1. Introduction
Methionine (Met) is one of the essential amino acids (EAAs) in animals and plays an important metabolic and functional role. It is also the most limiting amino acid (AA) that stimulates the growth of cashmere, wool and mohair fibre [
1,
2,
3]. When the supply of Met to animals is insufficient, it will limit the synthesis of body proteins and cashmere fibre keratin, leading to the supply of an AA surplus relative to the requirement [
3,
4]. Furthermore, surplus AAs are catabolized in the liver, and their nitrogen (N) content is converted into urea, with a large proportion being excreted via urine [
5]. The data from monogastric animals showed that insufficient Met in the diet can reduce the digestibility of protein in the small intestine and increase the faecal N output [
6]. The N in faeces and urine is gradually converted into nitrate, ammonia and nitrous oxide during the storage process, which contribute to air, soil and water pollution, have adverse effects on the environment and lead to global warming [
7,
8]. In addition to having adverse effects on the environment, the loss of dietary N in manure is also a significant economic loss for livestock farms, as protein is the most expensive dietary nutrient [
9]. Therefore, improving the N utilisation and environmental sustainability and enhancing the EAA supply, including Met, in low-protein diets to alleviate EAA deficiency is important in livestock production [
10].
Liaoning Cashmere goat is a major goat breed being raised in North-Eastern China, known for its high net cashmere rate and cashmere yield [
11]. The sulphur contents in cashmere fibre from goats is approximately 2.7–5.4%, and the higher the sulphur contents in cashmere fibre, the finer the cashmere fibre [
12,
13]. Therefore, sulphur-containing AAs are crucial nutrients for cashmere goats in harvesting high-quality fibres and achieving better growth performance [
14]. In North-Eastern China, crop straw (i.e., grazing) and a corn–soybean mixture are the main components of cashmere goat diets, which usually lack Met, and the offered protein frequently falls short of the recommended nutritional requirements [
15]. It has been observed that feeding rumen-protected Met (RPM) can improve the growth performance of goats [
16], milk yield of dairy goats [
17,
18] and wool characteristics of sheep [
19], as well as to improve N utilisation in goats [
16]. However, in relation to cashmere goats, the impact of RPM on performance and nutrient utilisation is currently poorly understood.
Unlike wool and mohair, cashmere fibres are produced seasonally [
20]. These changes vary along with nutritional conditions because of the seasonal nature of cashmere production [
21]. During the fast growth period of cashmere fibres (i.e., August to November), cashmere goats may have a relatively higher demand for protein and sulphur-containing AAs. These nutritional requirements drive the growth performance of cashmere goats [
13]. However, to date, there is limited information concerning the impact on bioavailability, e.g., plasma Met concentrations, N utilisation in cashmere goats, especially during the fast growth period of cashmere fibre. Therefore, this study aimed to determine the effects of RPM on the growth performance, nutrient digestion, manure N losses and plasma AA profiles of Liaoning cashmere goats during the fast growth period of cashmere fibre. The results we obtained will be potentially useful to the formulation of diets supplemented with RPM for maximising the production capacity of cashmere goats.
4. Discussion
Studies have shown that Met plays a specific role in protein metabolism and synthesis in the body and wool and is the primary limiting AA in furred animals, including cashmere goats [
12,
14]. Because Met deficiency may cause growth delays [
13,
25], this study investigated the effect of RPM supplementation in a Met-deficient diet on the growth performance and nutrient utilisation of cashmere goats. The findings indicated that ADG had a quadratic relationship with RPM level and increased 35.49 g/d for 2 g/kg RPM supplementation compared to the NC group during the 16-week feeding period. The ADG of 3RPM did not increase further relative to 2RPM, suggesting that 2RPM might have corrected the deficiency and helped goats meet the Met requirement. Moreover, the F/G was generally lower in the RPM groups than that in the NC group. The improvement in growth performance in goats may be due to increasing the Met availability in the small intestine to enhance the utilisation of N. Similar results were also reported, as feeding goats with 2.5 g RPM/kg DM improved their growth performance [
16]. Another study reported a significant improvement in ADG and F/G when actively growing Tan lambs were fed a low-protein diet supplemented with 1.5 g/d of RPM at a 1.0 and 0.8 voluntary intake [
26]. However, these findings are also inconsistent with some previous results. Obeidat et al. [
27] did not achieve a significant improvement in performance and F/G in ram lambs supplemented with different RPM doses. Similarly, no significant differences in performance were observed in Shaimi goats and beef steers supplemented with RPM [
28,
29]. Based on the inconsistencies between the present results and previous findings, the positive effect of RPM supplementation on growth performance in our study suggested that the cashmere goats required additional Met to address Met deficiency in order to maximise their body growth and cashmere fibre production. Considering the HSD results and the quadratic model of ADG and F/G, the 2RPM supplementation seemed to exhibit better results for growth performance.
Consistent with previous findings, the DM intake was similar across the different RPM doses, both during the digestibility determination period and the entire experimental period, as all parameters were kept similar, except for the RPM dose [
5,
16]. The apparent total tract digestibility of DM, NDF and N was improved using the RPM supplementation. Baghbanzadeh-Nobari et al. [
30] found that cows supplemented with DL-Met (1.2 g/kg DM) exhibited significantly increased apparent DM and ADF digestion. However, data on the relationship between nutrient digestibility and dietary RPM doses in ruminants remain limited. Early studies suggested that supplements with sulphur-containing AAs can better improve rumen microbial growth compared to supplements lacking these AAs [
31]. Improved rumen microbial growth should lead to more efficient DM, OM and NDF disappearance. However, in supplementation with Met from a rumen-protected source, some Met remains available to the rumen microbes [
32].
Nitrogen digestibility has often been assessed in ruminants receiving RPM in their diet. The data on dairy cows indicated that RPM supplementations at 0, 11.0, 19.3 and 27.5 g/d improved the apparent total tract digestibility of dietary N and decreased faecal N levels. Studies involving other species have indicated that Met supplementation can improve the AA balance in the diets, promote protein synthesis and enhance protein digestion and absorption by improving small intestine morphology and stimulating the production of digestive enzymes in the pancreas [
4,
16]. Similarly, urinary N excretion was not affected by RPM dose [
5]. Owing to a significant decrease in the manure N excretion, the N retention linearly increased with RPM dose, indicating an improvement in protein utilisation efficiency in the diet. Reductions in faecal N outputs improve the environmental sustainability of livestock farming, as the transformation of manure N into nitrate, ammonia and nitrous oxide contributes to soil, air and water pollution [
7,
33].
Plasma urea N concentration is often used to assess the nutritional status of livestock in terms of AAs, which reflects the efficiency in protein synthesis and the urea recycling rate [
34]. In our study, the plasma urea N concentrations significantly decreased, highlighting that whole-body N catabolism was affected by the RPM supplementation. The decrease in plasma urea N content indicates an increase in protein synthesis, whereas the opposite trend indicates an increase in protein catabolism and urea recirculating in the liver [
35]. In the present study, the plasma urea N concentrations in the 1RPM, 2RPM and 3RPM groups were lower than in the NC group (2.73, 1.96 and 2.91 vs. 3.99 mmol/L, respectively), with an average concentration of 2.53 mmol/L. All groups fell within the normal range [
34]. The regression analysis showed that the plasma urea N level was lowest when the RPM dose was 1.87 g/kg. Based on N utilisation resulting from the RPM dosage, the 2RPM treatment apparently satisfied the metabolic Met requirements of the investigated goats.
Plasma-free AA concentrations serve as an indicator of the amount of limiting AAs in the body, generally influenced by the composition of absorbed AAs [
36]. In this study, Met concentrations increased linearly and quadratically with RPM dose. The plasma Met concentration in treatment groups fed a diet containing 4.13–6.03 g of Met/kg DM exhibited 6.45–7.40 ng/mL, whereas the plasma concentration in the NC group exhibited 3.52 ng/mL. The elevated plasma Met concentration indicated that the rumen-protected technology had successfully presented Met to the small intestine and that Met had been released for absorption [
5]. Other studies have also reported an increased plasma Met concentration caused by supplementation with Met from rumen-protected sources [
37,
38,
39,
40,
41]. The availability of free AAs in the small intestine can positively influence blood AA concentrations [
32]; thus, the plasma Met concentration may be used to evaluate the duodenal flows of Met [
42]. When adequate energy is available, protein deposition shows a linear relationship with the supply of the most limiting AA, until another AA becomes more limiting [
43]. Inconsistent results have been reported regarding the ability of RPM supplementation to render Met available for absorption in the small intestine [
44,
45]. However, the present study showed that Met was absorbed, which affected the utilisation of other AAs. Based on the quadratic regression analysis, when the RPM dose was 2.4 g/kg DM, the plasma Met concentration was highest.
The plasma concentration of Lys, Phe, Leu and Ile decreased with the increase in RPM dose. Meanwhile, RPM supplementation had a negative impact on total plasma EAA, NEAA and TAA concentrations. These results suggested that the goats supplemented with RPM were more efficient in utilising Lys, Phe and two of the BCAAs (Leu, and Ile) than NC goats. These findings also indicated that cashmere goats supplemented with RPM had a more efficient utilisation of EAAs, NEAAs and TAAs, as the concentration of these AAs declined while the N retention increased on the 10th day. Toledo et al. [
43] also observed that feeding RPM in multiparous Holstein cows during the periparturient period increased the plasma Met concentration by about 38%; however, the concentrations of Leu, Val, Asp, Ser and Tyr were significantly decreased, and the TAA concentration tended to decrease. However, King et al. [
5] reported that supplementation of RPM (0, 11.0, 19.3 and 27.5 g/d) to a diet deficient in Met of dairy cows resulted in a linear increase in arterial concentration of Met, Lys, Try and Arg with RPM dose, while the BCAA concentration remained unchanged. The reason behind these inconsistent results is obscure. They could be attributable to differences in diet composition (i.e., some studies may have used treatment diets with insufficient Met, while others with sufficient Met), the dosage of supplementary RPM, and the age, breed and physiological status of the animals [
46]. But more elaborate studies should be conducted to better understand their dynamic roles in the growth performance of cashmere goats.