Choline and Methionine Protected in Crossbred Cows Diets

doi:10.21203/rs.3.rs-1983917/v1

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Choline and Methionine Protected in Crossbred Cows Diets

https://doi.org/10.21203/rs.3.rs-1983917/v1

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The objective was to evaluate the effects of providing protected choline and methionine on the productive performance and milk quality of crossbred cows. Twenty-four animals were used, distributed in randomized blocks according to the number of days in lactation. The treatments were: control, treatment with the addition of 50g of methionine (Methionine), treatment with the addition of 100g of choline (Choline) and finally the treatment with a blend of the two amino acids 100g of choline and 50g of protected methionine (COL/MET). At the beginning of the experiment, milk was weighed, which was repeated every 7 days to measure production. Every 21 days, individual milk samples were collected to prepare the physicochemical composition. In the weeks interspersed with the individual collection, the milk from each treatment was taken separately in order of treatment for the production of fresh Minas cheese and yield evaluation. Data were submitted to analysis of variance in a randomized block design, using a significance coefficient of 5.0% and Tukey's test to compare treatments. No significant differences were observed for production, milk composition and cheese yield, but the supplemented animals showed a greater persistence of production, with a reduction in production from the beginning to the end in the smaller experiment when compared to the control treatment. Additives did not increase milk production and economic yield of cheeses, but improved lactation persistence.

Feed conversion. Immunonutrition. Metabolic efficienc. Protected amino acid.

The use of amino acids (AA's), such as choline and methionine, both at the by-pass level, in the nutrition of dairy cows, has been gaining ground, especially in balanced diets for cows in the production phase. As methionine is the most limiting amino acid for milk production (VAILATI-RIBONI et al., 2017), along with choline, its precursor, measured by studies (AGUIRRE et al., 2015 and POTTS et al., 2020) as potent immunonutrients, capable of significantly improving the zootechnical indices of a dairy farm.

Methionine and choline have essential functions for both hepatic lipid metabolism and the immune system (ZHOU et al., 2016), and their availability during lactation is essential. Methionine is classified as a sulfur amino acid with a non-polar and aliphatic R group (FERREIRA et al., 2018), responsible for maintaining the serum values of metabolic amino acids, and choline is considered an important vitamin of the B complex, being an essential component of low-density lipoprotein molecules (AIRES et al., 2020), responsible for removing excess fat from the liver tissue, showing effectiveness as a treatment and prevention of hepatic liposis and ketosis.

Aguirre et al. (2015), in a study with simultaneous inclusions of protected choline and methionine, evaluated their effects on intake, lipid mobilization, milk production and composition in Holstein cows. In methodology, the authors blocked 24 cows in four treatments, t0 – control; t1–15 g/day of methionine; t2–15 g/day of choline and 15 g/day of methionine; t3–30 g/day of choline and 15 g/day of methionine. The results showed that lipid mobilization was higher for treatments in combination with additives (t2 and t3 (p = 0.0157)) and the values of dry matter intake (DMI), milk production and protein did not differ statistically between the two treatments. treatments, however, were affected by sampling times. However, there was a trend towards an increase in milk production and a reduction in solids, as we fed the additives together with a greater increase in choline (t3).

Potts et al. (2020) were able to assess the effect of choline and methionine inclusion contrasts for dairy cows fed singly or together, as well as the effect between primiparous and multiparous. For this, they selected 54 Holstein cows (25 primiparous and 29 multiparous). The authors found that milk production in the treatment that received choline without methionine increased by 8.7 kg/day in multiparous women, who also had increased milk fat concentration in the same treatment, with a tendency to increase the amount of fat produced. In contrast, methionine supplementation did not affect the production and overall fat concentration in milk for primiparous cows, however, choline supply increased milk production for primiparous cows by 3.5 kg/day, independent of the inclusion of methionine – the contrary to observations in multiparous cows.

The scientific support of studies with both additives, supplementing mostly herds of Holstein cows, is well known, however, there is little knowledge on the diets of crossbred cows managed under tropical climate conditions, being necessary to carry out more specific studies for the knowledge of its effects and therefore, the aim of the study was to evaluate the effects of providing protected choline and methionine on the productive performance and milk quality of crossbred cows (H:Z)

The Project was approved by the COMMISSION ON ETHICS IN THE USE OF ANIMALS (CEUA) of the Federal Institute of the Southeast of Minas Gerais under protocol Nº. 01/2022. It was carried out between 08/25/2021 and 10/20/2021, on the property “Sítio das Limeiras” located in the city of Juiz de Fora (Humaitá) - geographical coordinates: 21º46'20.8''S 43º29'30.2''W -, which is located in the dairy mesoregion, Zona da Mata Mineira.

Twenty-four animals were used, among crossbred cows and heifers (H:Z), with initial average milk production (MP) of 27.7 ± 4.47 liters/day, average weight (LW) of 540 ± 65 kg, days in lactation (DL) mean of 70.3 ± 40.2 days, mean body condition score (BCS) of 3.02 ± 0.46 and mean parturition order (OP) of 3.3 ± 1.25 deliveries. The animals were distributed in randomized blocks according to the DL and divided into 4 treatments: control, without addition of protected AA's, treatment with the addition of 50g of protected methionine per cow/day (Methionine), treatment with the addition of 100g of choline protected by cow/day (Choline) and finally the treatment with a blend of the two AA's, 100g of protected choline and 50g of protected methionine (COL/MET).

Once selected and identified by treatment, the animals formed a single production lot within the farm, receiving the same diet and management. 80% of the dietary requirement and AA's (Table 1) were offered individually in canzileiro troughs after morning and afternoon milking, which started at 5:20 am and 2:20 pm respectively. After consumption of the diet provided in the trough, they were released in a pasture covered by brachiaria brizantha cv. Marandu, for rest and to complement the fulfillment of 20% of nutritional requirements.

Table 1

– Diet in natural matter provided individually to the animals during the experiment.
Ingredients (kg)	Control	Methionine	Choline	COL/MET
Silage	25,27	25,27	25,27	25,27
Concentrate	8,02	8,02	8,02	8,02
Mineral	0,12	0,12	0,12	0,12
Methionine	-	0,05	-	0,05
Choline	-	-	0,1	0,1

The ration offered in the trough consisted of corn silage, concentrated with 28% of crude protein (CP) in natural and mineral matter, which were mixed by the forage wagon, sized in 1m³, for 5 minutes after adding the last ingredient and in sequence, unloaded. in drums to supply the troughs, differing between treatments only the inclusion of amino acids, which in turn, were placed separately and individually for each animal in its respective treatment.

To evaluate the daily milk production, the milk was measured weekly, individually, in meters attached to the milking machine. For the purpose of analyzing the physical-chemical composition: crude protein, fat, lactose, total dry extract, somatic cell count (SCC), milk urea nitrogen and casein contents, individual samples were collected every three weeks, packed in plastic bottles with preservatives (Bronopol®), kept between 2 and 6ºC, and sent to the Laboratory of Milk Quality Analysis of Embrapa Gado de Leite, in the city of Juiz de Fora – MG.

The BCS assessment was performed at the beginning of the experiment and weekly. The measurement took place after the afternoon milking, while the animals were in the trough, following the methodology of Ferguson et al. (1994), performed by two people, who judged from 1 to 5, extremely thin to extremely fat, respectively.

Every 21 days the animals were submitted to visual observation to evaluate the ingestive behavior. The animals were observed every 10 minutes, for 12 hours, from 6:00 am to 6:00 pm, to determine the time spent with water intake, diet consumption via trough, grazing, rumination or if they were idle.

The data on productive performance, body condition score and ingestive behavior were submitted to analysis of variance in a randomized block design. Data on milk production, milk composition, body condition score, ingestive behavior and cheese yield were submitted to the Tukey test at a 5.0% significance level, considering a significance of 5.0 to 10.0% as a trend through the SISVAR program (FERREIRA, 2011).

Milk production and composition

In the evaluation of the averages of milk production, from the beginning to the end of the experiment, as well as the averages per week and general (Table 2), there was no significant difference (P > 0.05) between treatments.

However, when evaluating the averages of the production difference between the beginning and the end of the experiment, there is a trend of difference (P = 0.0942) between the treatments, so, although we did not obtain the difference in the average production, it is possible to verify how much the animals that received the treatments with choline, methionine and the association between them maintain the production more constant and with less reduction when compared to the control.

The mean values of the physicochemical composition of milk from cows supplemented with choline and methionine (Table 3) did not differ statistically (P > 0.05). Similar results were also measured in the studies by Humer et al. (2019) and Zanton et al. (2014) and contrary to the results of Potts et al. (2020) and Toledo et al. (2017).

Table 2

– Mean values of initial production, production per week (1st to 7th), general and final average of dairy cows supplemented with protected choline and methionine.
Production(kg/dia)	Control	Methionine	Choline	COL/MET	SEM	P
Adaptation¹	29,08 a	27,13 a	28,05 a	26,36 a	1,3916	0,5606
1ª	28,2 a	27,33 a	28,01 a	26,83 a	1,4221	0,8978
2ª	26,07 a	25,64 a	28,83 a	27,42 a	1,5894	0,5023
3ª	28,06 a	27,14 a	27,86 a	28,45 a	1,7757	0,9615
4ª	27,77 a	26,72 a	26,73 a	27,46 a	1,2542	0,9089
5ª	26,54 a	24,83 a	26,42 a	25,38 a	1,3525	0,7730
6ª	27,09 a	26,94 a	28,39 a	26,52 a	1,5653	0,8494
7ª	25,32 a	25,13 a	25,39 a	26,45 a	1,4153	0,9109
General	27,38 a	26,33 a	27,46 a	26,8 a	1,3420	0,9248
Final²	26,20 a	26,04 a	26,89 a	26,48 a	1,4006	0,9749
Difference³	-2,88 a	-1,09 ab	-1,16 ab	0,12 b	0,771	0,0942

SEM – Standard error of the mean. P - Probability. ¹Average production from the fitting week to the first week after fitting. ²Average production of the last two weeks. ³Average difference of final production with the average of adaptation.

Table 3

– Mean values of the physicochemical composition of milk from dairy cows supplemented with protected choline and methionine.
Parameters	Control	Methionine	Choline	COL/MET	SEM	P
Fat (%)	3,5 a	3,07 a	3,59 a	3,51 a	0,3136	0,6311
Protein(%)	2,98 a	3,05 a	3,15 a	3,06 a	0,1195	0,7911
Lactose (%)	4,54 a	4,55 a	4,69 a	4,63 a	0,0913	0,6339
Casein (%)	2,36 a	2,41 a	2,52 a	2,40 a	0,0869	0,6136
Dry extract (%)	11,88 a	11,54 a	12,34 a	12,09 a	0,3973	0,2962
Urea (mg/dl)	23,05 a	23,39 a	22,97 a	22,54 a	0,7688	0,8904
SCCx1000 (CS/ml)	610,39 a	334,14 a	220,0 a	81,61 a	157,73	0,1538
SEM – Standard error of the mean. P - Probability.

Body condition score

Table 4

– Mean values of the initial, final body condition score, general average and difference between final and initial, of dairy cows supplemented with protected choline and methionine.
Parameters	Control	Methionine	Choline	COL/MET	SEM	P
Initial BCS	2,75 a	3,08 a	3,42 a	2,83 a	0,1848	0,0884
Final BCS	2,83 a	3,25 a	3,33 a	2,92 a	0,2753	0,5161
General	2,88 a	3,23 a	3,43 a	2,97 a	0,2157	0,2938
Difference	0,08 a	0,17 a	-0,08 a	0,08 a	0,1851	0,8101
SEM – Standard error of the mean. P - Probability.

The animals supplemented with protected choline and methionine did not differ statistically (P > 0.05) in ECC with the control treatment (Table 4). A result different from that found by Aguirre et al. (2015), who observed greater lipid mobilization – reduced BCS – for treatments in combination with additives (P = 0.0157).

Ingestive behavior

In the evaluation of the ingestive behavior results (Table 5), a statistical difference (P < 0.05) was observed for the average time spent in grazing (P = 0.02), total consumption (P = 0.03) and idle time (P = 0.01), with no difference (P > 0.05) for the times of trough, rumination and water intake.

Table 5

– Ingestive behavior in minutes, for 12 hours of evaluation, of dairy cows supplemented with protected choline and methionine.
Behavior	Control	Methionine	Choline	COL/MET	SEM	P
Grazing	105,0 a	134,17 ab	175,0 b	159,17ab	14,7180	0,0220
Trough¹	120,0 a	120,0 a	120,0 a	120,0 a	4,6398	1,0000
Total consumption²	225,0 a	254,17 ab	295,0 b	279,17ab	15,5665	0,0314
Rumination	147,5 a	145,83 a	148,33a	160,0 a	11,6884	0,8202
Water	19,17 a	20,0 a	20,0 a	17,5 a	2,6352	0,4988
Idle	328,33 a	300,0 ab	256,67b	263,33 b	15,0938	0,0142
SEM – Standard error of the mean. P - Probability. ¹Consumption in the trough. ²Sum of time grazing with consumption in the trough.

Milk production and composition

The animals that received the COL/MET treatment arrived with the average production at the end of the experimental period increased by 0.12 kg, differing from the control treatment by 3.0 kg, which totaled an extra production in the period of 56 days, of 168 kg of milk per treated cow and, consequently, these animals tend to close a lactation period of 305 days with higher milk production.

Such results are associated with the consumption of dry matter and the metabolic interaction between nutrients in the diet, since choline acts directly on hepatic metabolism, enhancing the functions of nutrient synthesis via the liver and methionine, in turn, manages to reach higher proportion in the intestine and promotes a better balance of metabolizable AA's in the intestine, which is confirmed in the study by Vailati-Rioni et al. (2017), who report that the Lysine:Methionine ratio that most closely resembles milk protein is 3:1, and diets based on soybean meal as a protein feed tend to lack methionine to meet this ratio, which is the most limiting amino acid for milk production.

However, the absence of differences in the results between the studies for milk composition between animals supplemented with protected choline and methionine can be explained by the absence of difference in the average milk production between the tested herds, certain that the higher the milk production, the smaller the percentage of solids in it and the greater the amount of solids produced.

Therefore, when evaluating the urea concentrations in milk, which comes from the excess nitrogen recycled by the ruminant, metabolized by the liver and partially excreted via milk (KOZLOSKI 2017), a considerably high average of 23 mg/ dl According to Siachos et al. (2019), values from 18 mg/dl of urea nitrogen in milk already demonstrate excess ammonia nitrogen in the rumen, as well as excess CP and/or lack of carbohydrates in the diet, factors that may justify the lack of results for the other characteristics evaluated, milk production and composition. This was due to the failure to fully meet the nutritional requirements via the trough, with the supplement being made on pasture, which in turn may have been affected by the high level of non-protein nitrogen depending on the time of year – rain and regrowth.

Body condition score

The variation in BCS is related to the amount of nutrients metabolized by the animal in relation to its maintenance, production and reproduction requirements (FERGUSON et al., 1994), so the reduction occurs when the diet neglects some nutrient.

However, among the factors that alter the nutritional requirement of the lactating cow, LW, MP, DL, it is observed the absence of significant differences between the treatments, which also justifies the maintenance of the BCS in the present study.

Ingestive behavior

The time spent in the trough was the same for all treatments because the animals received the same individual diet in canzileiro troughs in the same period after milking. Therefore, it is observed that the differences in the average time in minutes of grazing and total consumption, added the times in trough and grazing, of the animals supplemented with methionine, choline and with both AA's in relation to the control treatment increased by 29.17, 70.00 and 54.17 minutes respectively and with that, they spent less time idle.

This increase is justified in the attempt of the animals supplemented with the additives to balance the diet and meet the nutritional requirements, as they maintained a more constant milk production during the experimental period, as well as maintaining the BCS.

In view of the above, it is concluded that the supply of protected choline and methionine to crossbred cows does not alter milk production and composition, but improves lactation persistence.

Ethics approval: the procedures were evaluated and approved by the Ethics Commission on the Use of Animals of the Federal Institute of Southeast Minas Gerais—Campus Rio Pomba, under protocol no. 01/2022.

Consent to participate: not applicable.

Consent for publication: not applicable.

Conflict of interest: the authors declare no competing interests.

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Choline and Methionine Protected in Crossbred Cows Diets

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Abstract

Introduction

Material And Methods

Results

Milk production and composition

Body condition score

Ingestive behavior

Discussion

Milk production and composition

Body condition score

Ingestive behavior

Declarations

References

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Version 1