Effects of feeding empty fruit bunch extract on milk production and milk fatty acid profiles in goats

doi:10.21203/rs.3.rs-4975723/v1

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Effects of feeding empty fruit bunch extract on milk production and milk fatty acid profiles in goats

https://doi.org/10.21203/rs.3.rs-4975723/v1

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The effect of feeding empty fruit bunch extract on milk production and milk fatty acid profiles of lactating Saanen goats was evaluated. A total of four lactating Saanen goats (14 days in milk, 54.3 ± 9 kg of body weight) were subjected to a crossover design with a 17-d period (10-day adaptation period and 7-day for data and sampling period) with a 12-day washout period in between both periods. The treatment diets were; CON; (0% extract, basal diet of 60:40 of napier grass and concentrate) and EFB (5% OPEFB extract per DM basal diet). The 5% EFB extract in EFB diet did not affects milk yield, milk fat, milk total solid and milk solid non-fat compared to the CON diet. The milk protein content was significantly higher in EFB diet (P < 0.05) while milk lactose content was significantly higher in CON diet (P < 0.001) when compared to the others. Milk from EFB diet shows significantly higher (P < 0.05) in capric, lauric, myristic acid, and total SFA concentration as compared with milk from CON diet. Goat fed with EFB diet significantly reduced (P < 0.05) in stearic, oleic, total UFA, total MUFA concentration and UFA: SFA ratio in milk when compared with goat fed with CON diet. Meanwhile, the content of myristoleic acid, linoleic acid and PUFA: SFA ratio was similar between both treatment diets. Hence, the supplementation of 5% OPEFB extract in EFB diet did not alter milk production but the composition of the milk especially the profiles of milk fatty acid.

Goat

oil palm by-products

extract

milk

fatty acid

The oil palm empty fruit bunches (EFB) are an abundant and easily accessible agricultural by-product from the oil palm industry. However, the high fiber content in the EFB hinders the potential of utilizing EFB in animal feed. Hence, the usage of EFB in extract forms in animal feed to explore its effect in milk production of the animal as oil palm byproduct has been kwown to contain a broad range of bioactive polyphenols that could improve rumen fermentation. A previous study by Jaffri et al., (2011) showed that oil palm leaves are rich sources of polyphenols, meanwhile, Al-Jumaili (2018) identified 56 chemical constituents in his study of oil palm leaf methanolic extract characterization with linolenic acid, methyl ester was the major compound of interest contained in the oil palm leaf extracts. Whereas, n- hexadecanoic acid was the major compound of interest found in oil palm fruits, with oleic acid came as the second major compound of interest (Odion et al., 2020).

The changes in fatty acid profiles as a result of using oil palm-based wastes in animal feed are lacking, especially in milk and blood but not in rumen (Ebrahimi et al., 2015 ; Aiman-Zakaria et al., 2017; Chanjula et al., 2021), and meat (Ebrahimi et al., 2011 ; Ebrahimi et al., 2013 ). Nutritional properties of milk can be further improved through the increased essential fatty acids content in milk. In our preliminary findings, EFB has been shown to contain a few plant bio-active compounds that exhibit antioxidant and antimicrobial properties (unpublished). Incorporation of polyphenol-rich OPEFB extract in the diet may influence fatty acid and natural phenolic compound transfer from diet to milk, hence delivering value-added dairy to consumers. As of this moment, there is limited literature published on the usage of empty fruit bunch extract as animal feed in evaluating milk fatty acid profiles. Therefore, the objective of this study is to determine the effect of empty fruit bunch extract on milk production and milk fatty acid profile.

Extract preparations

Fresh fruit bunchs were collected from Eng Hong Palm Oil Mill Sdn. Bhd. (GPS coordinates: 2.8598918783, 101.476206779), an oil palm processing plant in Banting, Selangor, Malaysia. Upon retrieving the fresh fruit bunch of oil palm, bunches were shredded into small pieces approximately 2cm long and 1cm wide. Then, it was soaked in methanol at the ratio of 1:3 w/v for 72 hours, and was shaken for every hour prior to the collection of the methanolic extract and strained through a 4-layer muslin cloth before storage at room temperature. The methanolic extract was centrifuged at 4000 rpm for 20 minutes before storage at room temperature if the methanolic extract still contain large particles from the empty bunches. The methanolic extract were then evaporated under partial vacuum using a rotor evaporator (Heidolph, Germany). The concentrate is then stored at -80°C freezer. The oil palm empty fruit bunch methanolic extract is then used to prepare the treatment diets for in-vivo animal trials.

Animal, diet and experimental design

Four female Saanen goats with body weight ranges 54.3 ± 9 kg (mean body weight ± standard deviation) were randomly chosen to undergo a crossover experimental design in which each animal was assigned to their respective treatment sequence. Each of the treatment diets consisted of 4 animals. The treatment diets were; control diets (CON): Napier grass (NG) / commercial concentrates (C), (60:40) without EFB extract and empty fruit bunch diet (EFB): Napier grass (NG) / commercial concentrates (C), (60:40) with 5% EFB extract inclusion. The diet was formulated to meet animal requirements (NRC, 2007) and was fed at 3% of body weight in two equal portions at 0800 and 1700, daily. Animals were housed in individual pens with free access to clean drinking water and mineral blocks. The feeding trial started at day 14 post-partum. It consisted of 10 days of adaptation period followed by 7 days of the experimental period, with a washout period of 12 days to ensure no carry-over effect for the next period of trial. Milk was collected twice daily and the yield was recorded while samples of milk were kept for further evaluation of milk composition and fatty acid profile. Blood and rumen were collected on day 17, 2 hours post-feeding and were kept for further analyses. Rumen fluid obtained was added with 5% of trichloro-acetic acid (TCA) to the samples for ammonia-N determination in a 1:1 ratio while for volatile fatty acid content, 10ml of rumen samples were acidified with 2ml of 25% meta-phosphoric acid, prior storage at -20°C. The chemical composition of the treatment diets were analysed as per standard method by AOAC (1990).

Table 1

Chemical composition of the treatment diets.
Items	Treatment diets
Items	CON	EFB
Chemical composition (% of DM)
Gross energy (MJ/kg of DM)	3.68	3.70
Dry matter	90.3	89.8
Crude protein	13.8	12.9
Neutral detergent fibre	59.4	58.3
Acid detergent fibre	38.6	40.5
Acid detergent lignin	4.73	5.40
Ether extract	1.74	4.14
Fatty acids, g/100g of total fatty acid
C10:0, capric	0.65	0.60
C12:0, lauric	6.90	7.33
C14:0, myristic	2.25	2.39
C14:1, myristoleic	0.77	n.d.
C15:0,pentadecanoic	n.d.	0.68
C15:1, pentadecanoleic	1.22	1.12
C16:0, palmitic	14.65	15.94
C16:1, palmitoleic	0.76	0.93
C18:0, stearic	3.75	2.42
C18:1n-9, oleic	12.55	12.66
C18:2n-6, linoleic	37.14	36.41
C18:3n-3, α-linolenic	17.40	19.53
Total SFA	28.20	29.35
Total UFA	71.80	70.65
Total MUFA	15.30	14.71
n-3:n-6 ratio	2.27	1.88
UFA: SFA ratio	2.59	2.44
PUFA: SFA ratio	2.05	1.95
CON (0%, basal diet: concentrate + napier grass (40:60%)), EFB (5% OPEFB per DM diet). SFA (saturated fatty acid): C10:0, C12:0, C14:0, C15:0, C16:0, C17:0, C18:0). UFA (unsaturated fatty acid): C14:1, C18:1n-9, C18:2n-6). MUFA (Monounsaturated fatty acid): C14:1, C18:1n-9. PUFA (polyunsaturated fatty acid): C18:2n-6. UFA: SFA (unsaturated fatty acid / saturated fatty acid). PUFA: SFA (polyunsaturated fatty acid / saturated fatty acid).*n.d.: not detected

Milk composition

Milk samples from individual goat was used to determine major component of the milk which is protein, fat, lactose, total solid and solid non-fat content at the dairy laboratory, Animal Science Department, Universiti Putra Malaysia. Approximately, 20 ml of milk sample was incubated in water bath (Techne Tempette Junior TE-8J Water Bath) at 40°C for 5 min. Then, the evaluation of milk composition was done by using a milk analyser (Laporte and Paquin, 1999), Milkoscan (134 A/B series equipment, Foss Electric, Denmark).

Fatty acid analyses of feed and milk

The fatty acid of feed and milk were extracted according to the method of Folch et al., (1957) with minor modifications describe by Ebrahimi et al., (2011) by using chloroform: methanol (2:1) (v/v) containing butylated hydroxytoluene to prevent oxidation during sample preparations. The transmethylation was conducted by using 0.66 N KOH in methanol and 14% methanolic boron trifluoride (Sigma Chemical Co. St. Louis, MO, USA). The fatty acid methyl ester was separated using Agilent 7890A gas– liquid chromatography (Agilent Technologies, Palo Alto, CA, USA) using a 100 m× 0.25 mm ID (0.20 µm film thickness) Supelco SP-2560 capillary column (Supelco, Inc., Bellefonte, PA, USA) equipped with a flame ionization detector (FID). The split ratio was 10:1 after injection of 1ul of the FAME. The injector and detector temperature was programmed at 250°C. The column temperature program initiated runs at 140°C for 5 minutes, warmed to 200°C at 3°C/min, held for 7 minutes, warmed to 220°C at 2.3°C /min, and then held for 3 minutes for facilitate optimal separation. The peak identified through comparison of equivalent chain lengths with a reference standard (Supelco 37 Component FAME Mix; Sigma- Aldrich, Inc., St. Louis, MO, USA). The fatty acid concentrations are expressed as g/100 g of the identified fatty acid.

Statistical analyses

All statistical analyses were carried out using SPSS (IBM version 22) by using an independent t-test to examine the effect of EFB feed on milk composition, and milk fatty acids profile. Differences between treatment means were deemed significant when P values ≤ 0.05 were observed.

Chemical composition and fatty acid profile of the treatment diets

The chemical composition of the treatment diets are presented in Table 1. The offered diets contained 12.91–13.8 of CP content and 3.70–3.68 MJ per kg of DM, respectively. The treatment diets content of NDF, ADF and ADL was different for both treatment, where NDF was in lower value (58.3% of DM) in EFB diet than CON diet (59.4% of DM). As for ADF and ADL content of the diets, CON diet contains lower values of ADF and ADL, 38.6 and 4.73% of DM compared with the EFB diet, which was 40.5 and 5.40% of DM, respectively. Meanwhile, higher ether extract content of EFB diets (4.14% of DM) was observed compared to the CON diet (1.74% of DM). Table 1 also shows the fatty acid profiles of the treatment diets. The fatty acid profiles of CON diets shows EFB diet contained higher value of lauric (C12:0), myristic (C14:0), pentadecanoic (C15:0), palmitic (C16:0), palmitoleic (C16:1), oleic (C18:1n9), and α-linolenic (C18:3n-3) compared to CON diet. This is reflected in the total SFA of the EFB diet which is considerably higher content (4.08%) than the CON diet.

Milk yield and compositions

Table 2 presents the result of milk yield and milk composition of goats fed with and without EFB extract. Milk protein content of goats fed with EFB extract (EFB) shows higher value (P < 0.05) compared to goats fed with control diet (CON). Meanwhile, milk lactose content of goats fed with control diet (CON) showed higher amount (P < 0.05) compared to those fed with EFB extract diet (EFB). An increment of 4.1% in milk protein content and a 2.8% decreased in milk lactose content was observed in milk of goats fed with EFB extract diet. Yet, there are no significant differences (P > 0.05) observed in milk yield, milk fat content, milk total solid content, and milk solid non-fat content between treatment diets.

Table 2

Effect of EFB on milk yield and composition (mean ± SD).
Parameters	Dietary treatment		SEM	P value
Parameters	CON	EFB	SEM	P value
Milk yield (kg/day)	1.20 ± 0.23	1.12 ± 0.44	0.05	0.407
Milk composition
Fat (%)	4.11 ± 0.65	4.03 ± 0.98	0.07	0.612
Protein (%)	3.15 ± 0.27	3.28 ± 0.44	0.03	0.039
Lactose (%)	5.01 ± 0.20	4.87 ± 0.26	0.02	0.001
Total solid (%)	12.62 ± 0.87	12.47 ± 1.24	0.09	0.420
Solid non-fat (%)	8.70 ± 0.41	8.64 ± 0.61	0.04	0.505

CON (0%, basal diet: concentrate + napier grass (40:60%)), EFB (5% OPEFB per DM diet).

Milk fatty acid profiles

The results of milk fatty acid profiles are presented in Table 3. There is no significant changes in myristoleic acid, palmitic acid, and linoleic acid. Meanwhile, significant changes was recorded on milk fatty acid of capric acid, lauric acid, myristic acid, stearic acid, and oleic acid. EFB diet shows a significant increase in total SFA, with a significantly reduced total UFA and MUFA of the milk. Additionally, milk fatty acid ratio of UFA : SFA was significantly reduced while the ratio of PUFA : SFA of the milk was unaffected.

Table 3

Fatty acid composition of milk from goats fed EFB (mean ± SD).
Fatty acid, g/100g total fatty acid	Dietary treatment		SEM	P value
Fatty acid, g/100g total fatty acid	CON	EFB	SEM	P value
C10:0, capric	9.64	13.01	0.709	0.014
C12:0, lauric	7.35	9.50	0.550	0.048
C14:0, myristic	8.91	12.29	0.759	0.022
C14:1, myristoleic	7.53	6.62	0.655	0.495
C16:0, palmitic	26.09	28.67	0.724	0.075
C18:0, stearic	9.26	7.40	0.433	0.029
C18:1n-9, oleic	26.06	20.47	1.258	0.024
C18:2n-6, linoleic	8.31	8.03	0.582	0.818
Total SFA	61.17	70.28	1.990	0.019
Total UFA	38.83	30.88	1.960	0.039
Total MUFA	31.60	25.22	1.636	0.049
UFA : SFA	0.69	0.47	0.054	0.043
PUFA : SFA	0.15	0.13	0.013	0.381

CON (0%, basal diet: concentrate + napier grass (40:60%)), EFB (5% OPEFB per DM diet). SFA (saturated fatty acid): C10:0, C12:0, C14:0, C15:0, C16:0, C18:0). UFA (unsaturated fatty acid): C14:1, C18:1n-9, C18:2n-6). MUFA (Monounsaturated fatty acid): C14:1, C18:1n-9. PUFA (polyunsaturated fatty acid): C18:2n-6. UFA: SFA (unsaturated fatty acid / saturated fatty acid). PUFA: SFA (polyunsaturated fatty acid / saturated fatty acid).

It is widely known that the fat supplementation in the diet of dairy cows and ewes leads to lowered milk protein content and modifying coagulated properties, which this negative effect was not visible in goats (Chilliard and Bocquier, 1993) since most fat supplementation in goat results in increasing milk protein content, which concurred with the current study. The increment trend of milk protein content in this study was similar with a study by Zailan et al. (2023) in its study of feeding treated-EFB to the dairy goats, which significantly increase milk protein content. It has also been observed by Chilliard et al., (2003) in its review studies that the addition of fat to the goat diet not only causes no reduction in milk protein content, but an increase in 14 out of 20 reviewed trials. The lactose content is the only source of carbohydrates in milk (Chandan et al., 1992). Milk lactose content in the present study was in a similar range to the findings of Simos et al., (1991), Chandan et al., (1992) and Greyling et al., (2004). Milk lactose content is positively correlated with milk yield (r = 0.10) (Mioc et al., 2008) which is in line with the result of the present study where milk yield is decreased, albeit insignificant.

The current study results were similar to a previous study by Chamberlain and DePeters (2017) where high content of palmitic acid in milk was found and it was due to the high content of palmitic acid in the diet. The current study observed a significant increment of milk fatty acid occurs at medium-chained fatty acid, which is capric acid, lauric acid, and myristic acid that originated from de novo synthesis in the mammary gland where acetate and β-hydroxybutyrate are the major substrate sources. While acetate is the end-product of carbohydrate fermentation in the rumen, β-hydroxybutyrate is produced by the rumen epithelium from absorbed butyrate (Bernard, Leroux and Chilliard, 2008). This increment may be explained by both rumen acetate and butyrate concentrations in our unpublished study, although statistically insignificant, showing an increase in amount when EFB feed is given to the animal. This results in the total SFA increases in milk of EFB feed. As for LCFA in the milk, they are absorbed from blood plasma where the resources are from dietary lipid absorption and from body reserves mobilisation. In the current study, supplementation of the EFB feed has led to decreased concentration of stearic and oleic acid in the milk which suggests that the EFB extract diet improved energy density availability to animals. This is because when the energy balance is negative, animals mobilise lipids stored in adipose tissue, mainly in the form of non-esterified fatty acid (NEFA). Since ruminant adipose tissues are in high content of palmitic, stearic and oleic acid as described by Bas et al., (1987) in goat tissues, 59% of the variability in milk content of C18 and C18:1 is related to energy-balanced changes, in goats with differences in milk yield during the first four months of lactation, fed with a classic diet of hay and concentrate diet (Chilliard et al., 2003). A lower UFA: SFA ratio was observed in milk of EFB feed due to a higher proportion of SFA content in the milk. The ratio of UFA: SFA is used to reflect the hardness of butter which varies throughout lactation (Soyeurt et al., 2007), as the values increase rapidly to 2 up to 100 days in milk (DIM) and decrease slowly until 365 DIM, in cows. The changes appear due to the seasonal effect which is mainly due to changes in feeding especially in summer as stated by Soyeurt et al., (2007). The value of milk UFA: SFA ratio in this study (0.468–0.685) was similar to the findings of Correddu et al., (2016) in the study of dietary supplementation of grape seed and linseed on milk fatty acid of ewes. Additionally, PUFA: SFA ratio is another frequently used indices that was used to evaluate fatty acid profile related to cardiovascular health (Chen and Liu, 2020). Current study shows the ratio of PUFA: SFA was similar to the study by Mierliţă (2018) of dietary hemp supplementation on ewe’s milk where its PUFA: SFA ratio is in the range of 0.106–0.175, while PUFA: SFA ratio of this study was in the range of 0.126–0.150. Nevertheless, the factors that change the milk fatty acid which stems from rumen fatty acids are rumen biohydrogenation and fatty acid esterification (Lanier and Corl, 2015). Among the challenges that prevent easy incorporation of dietary fatty acids into milk are the fatty acids cellular uptake and the differences in fatty acids incorporation into milk as stated by Lanier and Corl (2015), although these received lesser research attention.

In conclusion, supplementation of the OPEFB extract at 5% can be employed in animal feed as it posed no adverse effect on milk yield, milk fat, milk total solid and milk solid non-fat. The 5% OPEFB extract supplemented in a high roughage diet increased the milk protein content and reduced milk lactose content while changing the composition of milk fatty acid profile towards a higher saturated fatty acid content. In general, EFB feed is a suitable feed to be incorporated in animal feed to produced milk with a wholesome dairy matrix.

Ethics approval:

The protocol for the animal study was reviewed and approved by the Animal Care and Ethics Committee of the Universiti Putra Malaysia (UPM/IACUC/AUP-R0048/2017).

Consent to participate
Not applicable.

Consent for publication
Not applicable.

Competing Interest
The authors declare no competing interests.

Funding

This study was supported by the Transdisciplinary Research Grant Scheme, Ministry of Higher Education, Malaysia [TRGS/1/2016/UPM/02/5/1].

Author contribution

All authors contributed to the study conception and design. NLAH conducted the experiment, data collection and data analysis. GYM validated the statistical analysis. NLAH wrote the first draft of the manuscript with the inputs from AAS, AQS, and GYM. All authors have read and agreed to the final manuscript.

Data availability

The associated author provides the datasets and resources upon reasonable request.

Code availability

Not applicable.

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Effects of feeding empty fruit bunch extract on milk production and milk fatty acid profiles in goats

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Version 1

Abstract

Introduction

Materials and methods

Extract preparations

Animal, diet and experimental design

Milk composition

Fatty acid analyses of feed and milk

Statistical analyses

Results

Chemical composition and fatty acid profile of the treatment diets

Milk yield and compositions

Milk fatty acid profiles

Discussions

Conclusion

Declarations

Ethics approval:

Consent to participate
Not applicable.

Consent for publication
Not applicable.

Competing Interest
The authors declare no competing interests.

Funding

Author contribution

Data availability

Code availability

References

Status:

Version 1

Effects of feeding empty fruit bunch extract on milk production and milk fatty acid profiles in goats

Status:

Version 1

Abstract

Introduction

Materials and methods

Extract preparations

Animal, diet and experimental design

Milk composition

Fatty acid analyses of feed and milk

Statistical analyses

Results

Chemical composition and fatty acid profile of the treatment diets

Milk yield and compositions

Milk fatty acid profiles

Discussions

Conclusion

Declarations

Ethics approval:

Consent to participate Not applicable.

Consent for publication Not applicable.

Competing Interest The authors declare no competing interests.

Funding

Author contribution

Data availability

Code availability

References

Status:

Version 1

Consent to participate
Not applicable.

Consent for publication
Not applicable.

Competing Interest
The authors declare no competing interests.