CADERNO DE CIÊNCIAS AGRÁRIAS
Agrarian Sciences Journal
Use of docosahexaenoic acid for lamb diet
Maria Júlia Pereira de Araújo
1
; Erica Beatriz Schultz
2
*; Thauane Ariel Valadares de Jesus
3
; Marina Elizabeth
Barbosa Andrade
4
; Luciana Melo Sousa
5
; Gilberto de Lima Macedo Júnior
6
DOI: https://doi.org/10.35699/2447-6218.2020.20203
Abstract
The purpose of this study was to evaluate the effects of docosahexaenoic acid in the concentrated acid on intake,
digestibility, metabolic profile and ingestive behavior of lambs diets. Five crossbred Dorper x Santa Inês lambs with
initial age and body weight average of six months and 33 kg, respectively, were used in the trial. The animals were
assigned into a 5×5 Latin square design and they received one of the following treatments: 0, 1.5, 3, 4.5 and 6 % of
microalgal product (Aurantiochytrium limacinum algae) or ALL-G Rich
®
in the concentrate. The diets were composed
of corn silage and concentrate offered twice daily. During the experimental period, feed, water, feces, blood, and
urine were sampled to calculate the intake and digestibility and to characterize the metabolic profile. The ingestive
behavior was measured during 24 hours on the last day each experimental period. The time spent on feeding, rumi-
nating and idle activities was measured. Regression analyses were performed considering 5 % of significance. There
were no difference on nutrients intake, digestibility, and ingestive behavior with the inclusion of ALL-G Rich
®
in the
diet (p>0.05). There was a significant effect on serum concentrations of cholesterol, low density lipoprotein, alkaline
phosphatase and gammaglutamyl transferase, however, all metabolites remained within the range recommended for
the animal category. Thus, ALL-G Rich
®
can be used up to 6% in the concentrate, on dry matter basis, of lambs diet
without affect intake, digestibility, metabolic profile, and ingestive behavior.
Keywords: Digestibility. Intake. Lipids.
Uso do ácido docosahexaenóico na dieta para cordeiros
Resumo
A proposta da pesquisa foi avaliar o efeito da inclusão de ácido docosahexaenóico no concentrado da dieta de cordei-
ros sobre o consumo, digestibilidade, perfil metabólico e comportamento ingestivo. Foram utilizadas cinco borregas
mestiças (Dorper x Santa Inês) com média de 6 meses de idade e 33 kg. Os animais foram distribuídos em delinea-
mento quadrado latino 5x5 sendo os tratamentos: 0; 1,5; 3,0; 4,5 e 6% de ALL-G Rich
®
no concentrado. As dietas
foram compostas de silagem de milho e concentrado ofertadas duas vezes ao dia. Durante os períodos de avaliação
foram mensurados e amostrados os alimento, água, fezes, sangue e urina. A partir dessas amostras foram calculados
o consumo e digestibilidade. O comportamento ingestivo foi realizado durante 24 horas no último dia cada período
experimental, mensurando o tempo gastos nas atividades de alimentação, ruminação e ócio. Análises de regressão
1
Universidade Federal de Uberlândia. Uberlandia, MG. Brasil.
https://orcid.org/0000-0002-7730-5828
2
Universidade Federal Rural do Rio de Janeiro. Rio de Janeiro, RJ. Brasil.
https://orcid.org/0000-0003-1916-2117
3
Universidade Federal de Uberlândia. Uberlandia, MG. Brasil.
https://orcid.org/0000-0003-3127-7697
4
Universidade Estadual Paulista. Jaboticabal, SP. Brasil.
https://orcid.org/0000-0003-3713-6377
5
Universidade Estadual Paulista. Jaboticabal, SP. Brasil.
https://orcid.org/0000-0003-1016-8248
6
Universidade Federal de Uberlândia. Uberlandia, MG. Brasil.
https://orcid.org/0000-0001-5781-7917
*Autor para correspondência: ericabeatrizschultz@gmail.com
Recebido para publicação em 17 de Abril de 2020. Aceito para publicação 25 de Agosto de 2020
e-ISSN: 2447-6218 / ISSN: 2447-6218 / © 2009, Universidade Federal de Minas Gerais, Todos os direitos reservados.
Araújo, M. J. P. et al.
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Cad. Ciênc. Agrá., v. 12, p. 01–08, https://doi.org/10.35699/2447-6218.2020.20203
foram utilizadas considerando 5% de significância. Não houve efeito significativo diante do consumo e digestibilidade
de matéria seca e dos nutrientes, tal como no comportamento ingestivo com a crescente inclusão de ALL-G Rich
®
na
dieta. Houve efeito significativo para as concentrações séricas de colesterol, lipoproteína de baixa densidade, fosfatase
alcalina e gamma glutamil transferase, porém todos os metabolitos mantiveram-se dentre da faixa recomendada para
categoria. Portanto, a ALL-G Rich
®
pode ser utilizada até 6% na matéria seca do concentrado em dietas de cordeiras
sem afetar o consumo, digestibilidade, o perfil metabólico e o comportamento ingestivo.
Palavras-chave: Consumo. Digestibilidade. Lipídeos.
Introduction
Lipids in the feed are energy source that de
-
creases increment caloric, improves energy efficiency,
and absorption of vitamins. The importance of lipids is
related to the fatty acid profile, where saturated fatty
acids lack double bonds and unsaturated fatty acids have
at least one double bond. Also, fatty acids can be divided
according to requirement and availability as essential and
nonessential fatty acids (Palmiquist & Mattos, 2011).
As examples of essential fatty acids, linoleic (ome-
ga-6) and linolenic (omega-3) acids are encountered
mainly in fish oil, canola, linseed, soy, and algae, which
has attracted attention for human health because of the
decrease in obesity, cardiovascular disease, and increase
in immunity (Calder, 2014). Also reported by Stark et
al. (2016) that animal producers enriched with omega-3
long-chain polyunsaturated fatty acids (LCPUFA), eico-
sapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic
acid (DHA, 22:6n-3) have been associated with a decrea-
sed risk of chronic disease, in particular cardiovascular
mortality and cognitive decline.
Conventionally, in ruminant nutrition is used
1 to 5% of lipids in the diet because more than 6% can
reduce microorganism access to fiber and exceed hydroly-
ze capacity, reducing intake and digestibility of the feed
(Palmquist & Jenkis, 1980). A ruminant particularity is the
biohydrogenation process in the rumen, which converts
unsaturated to saturated fatty acids. Thus, the offered
unsaturated source will be absorbed as saturated fatty
acids in the intestine (Church, 1993). The exceptions
that escape from this process is the polyunsaturated
fatty acids (PUFA) as docosahexaenoic (omega-3) and
eicosapentaenoic (omega-3) acids that are absorbed at
intestinal level (Ponnampalam et al., 2009).
Some studies using distinct sources of omega-3
in the ruminant feed showed a decrease in methane
production, improvement in animal health, meat and
milk fatty acids profile for human consumption without
damage animal’s performance (Boeckaert et al., 2008;
Pirondini et al., 2015; Thanh et al., 2018). Therefore,
considering the contribution for animals and human our
hypothesis is that the inclusion of docosahexaenoic acid
can alter intake, digestibility, ingestive behavior and the
metabolic profile of lambs.
The aim of this research was to evaluated the
effect of inclusion of docosahexaenoic acid on nutrient
intake, ingestive behavior, digestibility and metabolic
profile of lambs.
Material and Methods
All procedures were conducted in accordance
with the guidelines set out by the Brazilian College of
Animal Experimentation in the Code of Practice for the
Care and Use of Animal for Experimental Purposes, which
were approved by the Ethics Committee on Use of Ani-
mal for Research (CEUA) of the Universidade Federal de
Uberlândia under license number 183/2016.
Five lambs crossbred Dorper x Santa Inês with
an initial average age of 180 ± 8 days and body weight
(BW) of 33.59 ± 6.41 kg were assigned into a 5×5 Latin
square design. The animals were housed in individual
pens of 2 m
2
with individual feeders and drinkers accor-
ding to National Institute of Science and Technology
(INCT). In total, the trial lasted 75 days, divided into five
experimental periods of 15 days each one. An adaptation
period was performed from the 1
st
to 10
th
day of each
period and the collection period of feed, water, feces and
urine sampling was performed from 10
th
to 15
th
day. Also,
observation of ingestive behavior was performed from
14
th
to 15
th
day of each experimental period.
The diets were composed of corn silage, corn
meal, soybean meal, and mineral premix. The treatments
were characterized by the inclusion of 0, 1.5, 3, 4.5 and 6
% of the microalgal product (Aurantiochytrium limacinum
algae) or ALL-G Rich
®
in the concentrate. The ALL-G
Rich
®
composition are showed in Table 1.
The ration was formulated according to Nutrient
Requirements of Small Ruminants (NRC, 2007) for an
average daily gain of 200 g. Also, the diets were compo-
sed of 30 % of roughage and 70 % of concentrate. The
concentrate composition and chemical composition of
the experimental diets are showed in Table 2.
Araújo, M. J. P. et al.
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Table 1 – ALL-G Rich
®
chemical composition.
Item* Value Item* Value
Ethereal extract (%) 50 Glycerol (%) 3.81
Carbohydrates (%) 24.88 Monoglycerides (%) <1.0
Crude Protein (%) 19.22 Triglycerides (%) 85.80
Diglycerides (%) 4.69 Docosahexaenoic acid (%) 27.20
*The data were provided by Alltech
®
.
The silage and concentrate were weighted and
offered twice a day (08 h 00 min and 16 h 00 min) as
total mixed ration (TMR). The amount of supplied feed
was corrected to generate 10% of leftovers in the dry
matter basis. To adjust the amount of provided ration,
the animals were weighted in the first and last day of
each experimental period. In each experimental period,
samples of feed and leftovers were collected daily and
stored in plastic bags at -20°C. Also, the amount of total
feces was weighted and sampled daily in each experi-
mental period and stored in plastic bags at -20°C.
Table 2 The concentrate and chemical compositions of the experimental diets.
ALL G-Rich
®
(%)
Item 0 1.5 3 4.5 6
Corn meal 67.35 66.25 65.2 64.1 63
Soybean meal 30.45 30.05 29.6 29.2 28.8
ALL-G Rich
®
0 1.5 3 4.5 6
Mineral premix 2 2 2 2 2
Adsorbent 0.2 0.2 0.2 0.2 0.2
ALL G-Rich
®
(%)
Item 0 1.5 3 4.5 6
Crude protein (%) 16.55 16.55 16.55 16.55 16.55
Neutral detergent fiber (%) 19.94 19.76 20.18 19.78 20.13
Acid detergent fiber (%) 11.08 11.43 11.34 11.09 11.33
Ether extract (%) 1.60 1.65 1.67 1.69 1.79
Total digestive nutrients (%) 76.84 77.01 77.18 77.34 77.51
An amount of 6 liter of water per animal in a
bucket with capacity of 8 liter was offered once a day at
08 h 00 min. Meanwhile, one reference bucket of same
size with 6 liter of water was used to measure the daily
evaporation, thus, the water intake was daily measured.
Also, urine production were measured through graduated
test tube with precision of 20 mL in each experimental
period.
Samples of feeds, leftovers and feces were analy-
zed to determine the concentration of dry matter (DM),
ether extract (EE) and crude protein (CP) (AOAC, 1990).
The concentration of neutral detergent fiber (NDF) and
acid detergent fiber (ADF) were based on the description
of Van Soest et al. (1991).
The DM, EE, NDF, and ADF intake were calcu-
lated by difference between offered feed and leftovers.
The water intake was determined by difference among
offered, leftover, and evaporation. The DM and NDF
digestibility was estimated by difference among intake
and feces divided by intake. After quantification of urine
volume, a sample of 100 mL was collected to measure the
urine density, using Megabrix
®
manual refractometer.
Blood samples were collected on 11
th
, 13
th
and
15
th
day of each experimental period before first feeding
by jugular vein puncture with auxiliary vacuntainner tubes
without anticoagulant. After collection, the blood samples
were immediately centrifuged at 2.700×g for 20 min.
The plasma samples were pipetted and frozen at -18°C for
later analysis according to Russell et al. (2007) for total
protein, albulmin, globulin, uric acid, urea, creatinine,
cholesterol, triglycerides, low density lipoprotein, high
Araújo, M. J. P. et al.
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density lipoprotein, very low density lipoprotein, aspartate
aminotransferase, gammaglutamyl transferase, and alka-
line phosphatase using a commercial kit from Lab Test
®
in an automated biochemical analyzer (Bioplus
®
2000).
Feeding behavior such as eating, ruminating and
idling time of all ewes were evaluated on 14
th
to 15
th
day
of each experimental period in every five minutes during
24 hours, according to methodology proposed by Fischer
et al. (1998). The chewing time was calculated by sum
of eating and rumination times. The eating, rumination
and chewing efficiencies were calculated by dividing the
intake of dry matter by the total eating, rumination and
chewing times.
The statistical model was:
;
where:
= observation ijkl; = treatment
fixed effect i; = period fixed effect j; = animal
random effect k; = random error l. Comparisons
between ALL-G Rich
®
levels in the diets were conducted by
the sum decomposition of squares in orthogonal contrasts
to linear and quadratic effects with 5% of significance (p
< 0.05).
Results and Discussion
Intake of DM, DM in relation to body weight, EE,
NDF, and ADF were not different across the treatments
(p > 0.05) (Table 3). The ruminant intake is regulated
by a compilation of factors, for instance, physical (i.e. fill
ruminal), chemical (i.e. satiety), and psychogenic (i.e.
palatability) associated with diet composition (Mertens,
1994). Furthermore, inclusion of algae percentages sho-
wed no differences on chemical profile of the diets (isoe-
nergetic and isoproteic) (Table 2), leading to equivalent
nutrient intake between treatments.
Another fact related with no change in nutrients
intake can be the percentage of ether extract in the diets.
According to Palmquist e Jenkis (1980) the maximum
amount of lipids in the diet should be between 5% to avoid
negative effects on ruminal microbiota. Therefore, in our
experimental diets with 1.79% maximum of EE, there was
no influence on nutrient intake among treatments (Table
3). As reported by Thanh et al. (2018), where sunflower
and fish oil were used as sources of docosahexaenoic,
there was no difference in the nutrients intake because
the maximum percentage ethereal extract was 4.67 %,
no exceeding the recommendations.
In relation to lipid composition, the algae product
is rich in polyunsaturated fatty acids (PUFAS), as doco-
sahexaenoic acid, which according to Altomonte et al.
(2018) may have toxic effects on the ruminal microflora.
However, we emphasize that given the low inclusion
of ALL G-Rich
®
up to 6% in the concentrate, it did not
change intake and nutrient digestibility (Table 3).
The average values of DM and NDF intake were
1.27 and 0.594 kg.d
-1
, respectively, corresponding to 3.78
and 1.76 % of body weight, respectively. The average
of NDF and NDF in relation to BW were higher than
reported by Mertens (1997), which suggest that NDF
intake can reach 1.20% of BW, demonstrating that in
this study intake was not restricted by rumen fill. Howe-
ver, the average of DMI (1.0 to 1.30 kg day) and DMI
in relation to BW (2.97 to 3.87 % of BW) are within
the recommended for lambs according to NRC (2007).
Thus, we can conclude that despite NDF intake, %NDF
of BW was beyond suggested, do not observing intake
restriction.
According to Forbes (1996), intake regulator
factors operate in an integrated process, therefore, DM
intake was not limited by physical effects because diet
digestibility showed an average of 83.45%. Diets with
this percentage of digestibility is considered highly fer-
mentable and with high production of volatile fatty acids,
leading consequently to the interaction of physical and
osmotic intake controls. This control interaction happened
in all treatments, therefore, increasing ALL G-Rich
®
in
the concentrate did not change DM and NDF digestibility
(p >0.05) (Table 3).
The docosahexaenoic intake in the treatments
was estimated as 0, 1.5, 3, 4,5 and, 6 % of ALL G-Rich
®
,
being 3.78, 7.23, 10.96 and 13.70 g d
-1
, respectively.
Thus, ALL G-Rich
®
up to 6% with 13.70 g d
-1
as a source
of docosahexaenoic was used without change the nu-
trients intake and digestibility. This response opposes
that reported by Palmquist e Mattos (2011) and Borghi
(2018), it’s because these studies increased algae and
lipids more than our used, nearly 6%, thus, when we
increase PUFA sources in the ruminant diet the DM and
NDF digestibility decrease due to toxicity to ruminal
microbiota, hampering fiber degradation. The results of
these studies show that the unsaturated lipid can dama-
ge rumen, reducing DM and DND. However, the levels
included in our study were not detrimental to the dry
matter intake and digestibility.
Similarly to nutrients intake, there were no sig-
nificant differences among treatments for water intake
and water losses (p > 0.05) (Table 4). The water intake
and urine density in relation dry matter intake did not
change (p > 0.05) (Table 4). The average of water intake
and urine volume was 4.97 and 1.51 l d
-1
, respectively.
The water intake was 27.11 % higher to the calculated
by the equation proposed by Forbes (1968), which NRC
(2007) recommends. However, this equation is a general
requirement for all ewes class, and our study used growing
lambs. Thus, it is still necessary specific equations to all
small ruminants classes. The response of water is also
related to diet profile, which was composed of 70% of
Araújo, M. J. P. et al.
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concentrate containing little humidity, increasing intake
requirements of water. Urine volume and urine density
are within the range describe by Reece (2006) of 100
to 400 ml per 10 kg of BW and 1.015 to 1.045 g ml
-1
(Carvalho, 2008).
Table 3 – Digestibility and nutrients intake of lambs with levels ALL G-Rich
®
in the concentrate.
ALL G-Rich
®
(%) p-values
Item 0 1.5 3 4.5 6 SEM L Q
Dry matter intake (kg d
-1
) 1.35 1.27 1.28 1.27 1.20 0.053 0.157 0.992
Dry matter intake (kg d
-1
BW) 3.37 3.14 3.26 3.17 3.01 0.094 0.197 0.915
Ethereal extract intake (kg d
-1
) 0.032 0.031 0.031 0.030 0.032 0.001 0.958 0.234
Neutral detergent fiber intake (kg d
-1
) 0.591 0.48 0.591 0.511 0.573 0.026 0.974 0.303
Acid detergent fiber intake (kg d
-1
) 0.204 0.186 0.231 0.21 0.219 0.012 0.504 0.902
Dry matter digestibility (%) 83.39 83.53 81.31 84.45 84.06 0,009 0.399 0.635
Neutral detergent fiber digestibility (%) 65 60.79 67.29 68.14 62.14 2.735 0.625 0.982
BW: body weight; SEM: standard error of the mean; L: linear effect; Q: quadratic effect.
Table 4 – Intake and loss of water of lambs with levels ALL G-Rich
®
in the concentrate.
ALL G-Rich
®
(% of concentrate) p-values
Item 0 1.5 3 4.5 6 SEM L Q
Water intake (l d
-1
) 4.69 5.28 5.16 5.11 4.61 0.222 0.663 0.149
Water / Dry matter intake (l kg d
-1
) 3.43 4.65 4.2 4.19 4.13 0.303 0.386 0.084
Urine volume (l d
-1
) 1.528 1.523 1.737 1.292 1.474 0.188 0.771 0.744
Urine density (g ml
-1
) 1.022 1.021 1.014 1.025 1.020 0.002 0.874 0.697
SEM: standard error of the mean; L: linear effect; Q: quadratic effect.
In relation to metabolic profile, there was no
significant differences on total protein, albumin, globulin,
uric acid, urea, creatinine, triglycerides, very low den-
sity lipoprotein and high-density lipoprotein as increase
docosahexaenoic increased in the diet (p> 0.05) (Table
5). However, cholesterol and low-density lipoprotein
showed a significant (p<0.05) quadratic behavior as
increased feeding levels of ALL G-Rich
®
were added in
the concentrate (Table 5). All energy and protein meta-
bolites values are between reference values.
The metabolic profile is used to access and prevent
metabolic and nutritional disorders in animal production,
mainly, when unusual products are included in the diet.
Protein metabolites: total protein, albumin, globulin, uric
acid, urea, and creatinine are positive correlated with
dietary protein intake, rumen degradability, and dietary
amino acid composition (Puppel e Kuczynska, 2016).
As the diets were formulated to be isoprotein, there was
no difference between treatments for concentration of
protein metabolites.
The diet balance is verified through the urea
concentration. According to Wittwer (2000), blood urea
reflects the ammonia generated in the rumen by the
degradation of the nitrogen compounds that were not
converted into microbial protein. Thus, ammonia crosses
the rumen wall where it is transformed into urea in the
liver with high energy expenditure. Subsequently, this
urea can be eliminated by urinary excretion or milk (in
the case of lactating animals), and return to the rumen
via salivation or perfusion in the ruminal wall.
The urea values remained within the recommen-
ded range with inclusion of ALL G-Rich
®
. Treatments did
not affect the synergism of the degradation of nitrogen
and carbohydrate sources in the ruminal environment,
leading to no ammonia escape and subsequent no unne-
cessary energy expenditure for the animals. Therefore, all
protein metabolites remained within the normal range of
protein metabolism as recommended by Varanis (2018)
for growing lambs.
Among energetic metabolites, there were reduc-
tion in cholesterol and low-density lipoprotein concen-
trations up to 4%, and an increase in concentration with
6 % of ALL G-Rich
®
.
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Table 5 – Energetic and protein metabolites of lambs with levels ALL G-Rich
®
in the concentrate.
ALL G-Rich
®
(%) p-value
Item 0 1.5 3 4.5 6 SEM RV L Q
Total protein (mg dL)* 7.44 7.98 8.34 7.06 8.2 0.021 3.1 to 11.4 0.258 0.893
Albulmin (mg dL)* 3.49 3.85 3.29 3.18 2.86 0.001 1.12 to 5.38 0.398 0.748
Globulin (mg dL)** 3.94 4.45 5.04 3.88 5.34 0.148 3.5 to 5.7 0.473 0.698
Uric acid (mg dL)* 0.48 0.46 0.5 0.44 0.48 0.108 0 to 2.9 0.597 0.259
Urea (mg dL)* 48.56 46.46 57.76 46.78 48.44 0.225 12.8 to100 0.148 0.123
Creatinine (mg dL)* 0.74 0.72 0.74 0.72 0.74 0.001 0.40 to 1.80 0.163 0.543
Cholesterol (mg dL)
1
* 52.56 51.76 53.5 49.22 55.78 0.196 15 to 139.9 0.362 0.042
Triglycerides (mg dL)* 25.82 24.78 21.92 19.56 20.82 0.207 5 to 78 0.589 0.969
Low density lipoprotein (mg dL)*
2
21.22 11.12 11.95 8.12 14.85 0.366 0.80 to 83.36 0.093 0.001
High density lipoprotein (mg dL)* 37.56 41.64 40.04 37.18 36.76 0.284 13 to 79 0.111 0.473
Very low density lipoprotein (mg dL) * 5.16 4.95 4.38 3.91 4.16 0.189 1 to17.4 0.589 0.969
*RV: reference value accord to Varanis, (2018); **RV: reference value accord to Kaneko, (2008); SEM: standard error of the mean; L: linear effect;
Q: quadratic effect. ¹Y = 53.02 - 1.39X + 0.27X²; R² = 30.06%; ²Y = 20.74 - 6,56X + 0.92X²; R² = 86.20%
The ALL G-Rich
®
is rich in PUFA’s, which is
biohydrogenated in the rumen. When the docosahexae-
noic acid is biohydrogenated, it is converted to a long
saturated long chain fatty acid, the docosonaic acid (C
22:0) (Aldai et al., 2018). Thus, the increase in serum
cholesterol concentration may occur due to the high
demand for digestion, absorption, and transport of lon-
g-chain fatty acids (Freitas Júnior et al., 2010).
In this context, we call attention to the reduction
of low-density lipoprotein concentration (LDL) up to 4%
of ALL G-Rich
®
in the diet, the LDL acts on the transport
of cholesterol from the liver to the tissues. The increase
in cholesterol is indicative of an increase in energy me-
tabolism in the liver (NDLOVU et al., 2007).
The reduction of LDL concentration may occur-
red due to the greater use of fat from ALL G-Rich
®
in
the diet as energy source, which leaded to less need for
fat mobilization from the liver to the tissues. This is also
favorable because according to Brás et al. (2014) it leads
to a lower probability of developing health problems such
as cardiovascular diseases. It is worth mentioning that,
although there were differences in cholesterol and LDL
between treatments, all energy metabolites were within
recommended range to the animal category according
to Varanis (2018).
In relation to hepatic metabolites, there were
a quadratic and linear effect for alkaline phosphatase
and gammaglutamyl transferase (p<0.05), respectively.
However, means values of aspartate aminotransferase
were not affected by the increase DHA in the diet.
Table 6 – Hepatic metabolite of lambs with levels ALL G-Rich
®
in the concentrate.
ALL G-Rich
®
(%) p-value
Item 0 1.5 3 4.5 6 SEM RV* L Q
Aspartate aminotransferase (U L) 161.24 214.04 357.84 269.02 199.3 12,36 47 to 353.5 0.256 0.445
Alkaline phosphatase (U L)
3
283.98 328.18 347.56 311.62 328.16 25,36 58 to 727.7 0.398 0.025
Gammaglutamyl transferase (U L)
4
62.66 80.16 97.2 87.72 94.52 4,23 31 to154 0.558 0.002
*RV: reference value according to Varanis, (2018); SEM: standard error of the mean; L: linear effect; Q: quadratic effect;
3
Y = 289.73 + 25.86X -
3.51X²; R² = 61.48%;
4
Y = 70.19+ 4.75X; R² = 66.17%.
According to Dallago et al. (2011), elevated
concentrations of alkaline phosphatase, gammaglutamyl
transferase and aspartate aminotransferase are related
to hepatic injuries. When hepatic injuries happen, hepa-
tocellular damage overflows the hepatocytes by raising
their serum concentration. Although the values changed
with increase DHA in the diet, all metabolites hepatics
remained within reference intervals for lambs, showing
that there is no risk of intoxication when ALL G-Rich
®
is
used as source of DHA in lambs diet.
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In relation to animal behavior, the time lambs
spent in feeding, rumination, idle, and chewing as well
as efficiency times of these activities were not different
between treatments (p > 0.05) when added up to 6 % of
ALL G-Rich
®
in the concentrate (Table 7). Mean values
of feeding, rumination, idle and chewing were 191.8,
349.4, 898.8 and 541.2 min d
-1
, respectively.
The ingestive behavior is controled by several
factors, for instance, nutrients intake, particle size and
nutrients digestibility (Van Soest, 1994). We did not
observed changes in this factors among treatments, thus,
increased levels of ALL G-Rich
®
in the experimental diets
did not change the time spent in feeding, rumination,
idle and chewing activities, and consequently on their
efficiency. We call attention to the value of ruminating
time that was lower than showed by Van Soest (1994),
which reports a time of 360 to 480 min day
-1
rumina-
ting. However, the diet composition of this study had
high proportion of soluble carbohydrates. The values
of feeding, ruminating and idle activities of 3.19, 5.82
e 14.98 h day
-1
were similar as those reported by Silva
(2018) of 4.76, 5.98, 13.26 h day
-1
, respectively, using
up to 4% algae meal in lamb diet.
Table 7 – Ingestive behavior of lambs with levels ALL G-Rich
®
in the concentrate.
ALL G-Rich
®
(% of concentrate) p-values
Item 0 1.5 3 4.5 6 SEM L Q
Feeding time (min d
-1
) 187 188 197 195 192 6.524 0.712 0.627
Rumination time (min d
-1
) 347 352 351 368 329 8.804 0.761 0.375
Idle time (min d
-1
) 906 900 892 877 919 12.08 0.396 0.971
Chewing time (min d
-1
) 534 540 548 563 521 12.08 0.760 0.368
Feeding efficiency (g of DM min
-1
) 7.3 6.74 6.76 6.76 6.52 0.345 0.242 0.689
Rumination efficiency (g of DM min
-1
) 3.96 3.6 3.69 3.53 3.68 0.162 0.460 0.452
Chewing efficiency (g of DM min
-1
) 2.56 2.33 2.37 2.29 2.33 0.102 0.249 0.391
SEM: standard error of the mean; L: linear effect; Q: quadratic effect.
Conclusion
The ALL-G RICH
®
is viable to use on diets for lambs in
a level up to 6%, which represents up to 13.70 g d
-1
of
docosahexaenoic acid intake. This percentage of inclu-
sion do not affect ingestive behavior, digestibility, and
nutrient intake, also, maintains metabolic profile within
the recommended range for the animal category, increases
cholesterol and reduces LDL concentration.
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