Heliyon 6 (2020) e05831

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Heliyon
journal homepage: www.cell.com/heliyon

Research article

Effects of chemical silage of red tilapia viscera (Oreochromis spp.) as a source

of protein on the productive and hematological parameters in isa-brown
laying hens (Gallus gallus domesticus)
~o F b, Jose E. Zapata M a, *
Yhoan S. Gaviria G a, Luis F. London
a
Nutrition and Food Technology Group, Universidad de Antioquia, Medellín, Colombia
b
Politecnico Jaime Isaza Cadavid, Medellín, Colombia

A R T I C L E I N F O A B S T R A C T

Keywords: This paper evaluates the inclusion of chemical silage from viscera of red tilapia (Oreochromis spp.) in diets of Isa-
Chemical silage viscera Brown laying hens (Gallus gallus domesticus), and its influence on productive variables and hematological pa-
Red tilapia rameters. A total of 56, 16-week-old laying hens were randomly divided into two groups (one per diet), which in
Blood chemistry
turn were subdivided into 7 replicates of 4 birds each. All test groups were fed for 16 weeks. During this period,
Fish meal
Soybean meal
the evaluation of the productive variables was carried out, and at the end, random blood samples were taken from
Productive parameters 3 birds per diet. The results indicated that the inclusion of chemical silage from red tilapia viscera with a pro-
portion of 17.18% dry matter, does not present statistically significant differences in the productive variables
which were evaluated (p > 0.05) with respect to the control. Moreover, the chemical silage did not modify the
hematological parameters and blood the chemistry in the hens. This allowed us to conclude that silage can be used
as a substitute for conventional protein raw materials such as fishmeal and soybean meal in the preparation of
diets for laying hens, without altering their productive performance.

1. Introduction ecosystems at different levels. This can generate problems in ecosystems

such as biomass reduction, the density and diversity of benthos and
In recent years, human beings have acquired an advanced knowledge plankton, as well as the modification of natural trophic networks. These
about the health benefits associated with eating fish. This has led to a residues can be used for animal feed, to produce fishmeal, fats, or fer-
permanent growth of aquaculture production throughout the world. In tilizers, among others (Chalamaiah et al., 2012).
2016, global fish production reached 171 million tons, of which 86% was One of the most widely used strategies for nutrient recovery from
for direct human consumption (FAO, 2018). In 2017, the Colombian fish by-products consists of adding acids, enzymes, or lactic acid bac-
aquaculture sector underwent a growth in production of 9% with a total teria to raw fish, or parts of it. This is done in order to cause the pH to
of 120,230 tons, of which the red tilapia species held the most significant drop and generates a liquefaction of the mass by effect of endogenous
contribution with 62%, followed by white cachama at 16%, silver tilapia enzymes which act under acidic conditions (Fernandez Herrero et al.,
at 14%, trout at 5%, and a 3% of remaining species (Ministerio de 2013). This process is called silage (Suarez et al., 2018). The product
Agricultura, 2017). This growth has been followed by an equivalent obtained from said process is a grayish-brown colored, semi-liquid,
volume of waste, which can generate significant environmental impacts, pasty substance, with a characteristic fishy odor. The product provides
given that it represents between 60-70% (w/w) of total production partially hydrolyzed proteins, with nutritional properties which are very
(Martínez-Alvarez et al., 2015). These residues are mainly composed of similar to those of the fish (Botello, 2010) they come from. These
fillet remains (15–20%), skin and fins (1–3%), bones (9–15%), heads proteins could improve the digestibility of diets in animal feed, which is
(9–12%), viscera (12–18%) and flakes (5) (Martínez-Alvarez et al., one of the factors that positively influences growth. The poultry sector is
2015), which, despite being important sources of protein, are most often possibly the fastest growing and the most flexible of all the livestock
discarded without any attempt at reutilizing them. However, due to how sectors. Mainly driven by a strong demand, this sector has been
rich this waste is in terms of nutrients, its inadequate disposal can affect expanded, consolidated and globalized in the last 15 years in countries

* Corresponding author.
E-mail address: edgar.zapata@udea.edu.co (J.E. Zapata M).

https://doi.org/10.1016/j.heliyon.2020.e05831
Received 20 May 2020; Received in revised form 17 July 2020; Accepted 21 December 2020
2405-8440/© 2020 Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Y.S. Gaviria G et al. Heliyon 6 (2020) e05831

of all income levels (FAO, 2013). However, the high cost of poultry 2.4. Determination of productive variables
feeding is one of the main problems that plagues the production chain,
since approximately 95% of the chain is used to satisfy protein and Both diets were evaluated for weight gain by performing weekly
energy needs. Fishmeal and soy meal are the main sources of protein weighing rounds of the fasted animals with a 1g precision analytical
and corn flour is the main source of energy (Ravindran, 2013). The balance (TxB220-1L from Shimadzu, Japan). Eggs were collected,
objective of this paper was to evaluate the effect of the inclusion of counted and weighed daily for 13 weeks to determine the laying per-
chemical silage made from red tilapia (Oreochromis spp.) viscera, in the centage and their average weight.
elaboration of diets for Isa Brown laying hens. The study takes the
productive variables and hematological parameters into account in 2.5. Blood chemistry and hemogram
order to evaluate the effects of this dietary modification.
The complete hemogram was performed using the methodology
2. Material and methods proposed by Clark et al., in 2012, which counts red blood cells or
erythrocytes, white blood cells or leukocytes, and thrombocytes or
2.1. Handling of the materials platelets, which are all structures produced in the bone marrow by the
cytoplasmic fragmentation process (Clark et al., 2012). Hematological
The red tilapia (Oreochromis spp) viscera were supplied by the El analyses are among the methods which may contribute to the detection
Gaitero fish farm, located in the municipality of San Jer onimo, of some changes in health status and can be a useful aid for diagnosis
Antioquia-Colombia. Initially, the viscera were degreased, for which they diseases in birds (Schmidt et al., 2009). Blood samples were collected
were brought to 67
C for 30 min, with a subsequent drop to 45
C and from the wing vein, with or without anticoagulants, at from the 32th
the fat was decanted- After this, they were frozen (at -18
C) for 24 h. This week of age from all of the groups. Plasma samples were analyzed for
allowed the separation of the oil from the aqueous-protein phase, since total cholesterol, total protein, triglycerides, alkaline phosphatase and
when phases solidify they do so separately (Arias et al., 2017). Then, the the enzyme alanine aminotransferase (ALT) concentrations.
lipid phase was detached from the frozen aqueous-protein phase, by
means of a cross-section, and the aqueous-protein phase was stored for 2.6. Statistical analysis
the silage. The aqueous-protein phase was homogenized with a crushing
mill. Then, it was mixed with 0.03% sulfuric acid at 97% (Merck, Ger- The data collected was evaluated at a confidence level of 95% (P <
many) and 1.16% formic acid at 85% (Merck, Germany) to start the 0,05) by means of the hypothesis test to determine the difference in
hydrolysis and acidification process. The BHT antioxidant was then means using Fisher's LSD (Least significant difference) test with stat-
added (Butyl hydroxy toluene) (Tecnas SA, Colombia) along with an graphics centurion XVI software. A Completely Randomized Design was
antifungal - potassium sorbate (Tecnas SA, Colombia). Finally, the used in this study. There were seven replications for each treatment, and
mixture was stored at room temperature for a minimum time of 8 days. four layer hens in each replication. The treatments were 0 (control), and
During this period the product was homogenized by stirring, and a pH 32% silage of the viscera red tilapia substitution of basal diet.
reading was taken every two days until it stabilized.
2.7. Ethical approval

2.2. Poultry feed The experiments carried out in this manuscript were submitted and
approved by the ethics committee for experimentation with animals of
Fifty-six, 16-week-old Isa Brown laying hens were randomly divided the University of Antioquia - Colombia.
into two equal groups called the control group (CT) and the silage group
(ENS), which were fed from week 16 to week 32. The isoproteic diets 3. Results and discussion
were prepared following the nutritional requirements established by
Rostagno et al. (2011) (Table 3). The CT group was fed with the diet 3.1. Proximal composition of the silage
without the viscera silage, and the silage group (ENS) was fed with the
diet of which 32% of the protein was from the viscera silage. The hens Table 1 shows the bromatological composition of the viscera and the
were fed twice a day (morning and afternoon) with a 90 g/day ration for silage obtained from them. The viscera showed the levels of protein and
each bird, and a constant supply of water. fat typical to this species (Arias et al., 2017; Suarez et al., 2018). How-
ever, the values tended to be higher than those reported for acidic silage
from freshwater fish residues, with a dry basis protein of 44.38% (Vidotti
2.3. Physicochemical and microbiological analysis et al., 2003) and that of biological silage of tuna (Thunnus albacares) with
30.52% crude protein and 14.25% lipids (Spanopoulos-Hernandez et al.,
The physicochemical characterization of the silage was carried out in 2010).
accordance with the provisions of the AOAC (Association of Official In addition, the silage obtained showed an increase in protein content
Analytical Chemists). The moisture content was determined using the and a decrease in the level of fat, which is due to the degreasing process
methodology established in standard 930.15 (AOAC, 2000b), by drying to which the viscera were subjected. Said process is necessary to be able
the sample at 105
C for 8 h. The protein was determined by the Kjeldahl to balance diets such as the ones which were evaluated. A similar
method (standard 954.010) (AOAC, 2000a). The ashes were analyzed reduction was reported by Ramírez et al. (2013) authors who managed to
according to standard 942.05 (AOAC, 1995). Fat analysis was conducted reduce the ether extract from 24.5% to 14.5% on a wet basis.
according to standard 920.39 (AOAC, 2003). The carbohydrate deter- In this study, the product presented the appropriate characteristics of
mination was performed by subtracting one hundred percent of the an optimal silage: the substance was a grayish-brown colored, semi-
previously mentioned components from the sample (Spanopou- liquid paste with a characteristic fishy odor (Botello, 2010; Suarez
los-Hernandez et al., 2010). et al., 2018). The product's protein and fat levels allowed it to be an ideal
The microbiological analyzes were carried out according to the substrate for animal feed, since it provides partially hydrolyzed proteins,
Colombian Technical Standard NTC 3688 (ICONTEC, 1999), which with nutritional properties very similar to those of the original fish
prescribes the analysis of aerobic mesophylls, total and fecal coliforms, (Botello, 2010). This helps improve the digestibility of animal feed,
Salmonella spp., Clostridium spore reducing sulfites and molds and yeasts which is one of the factors that positively influences productive variables.
for these types of substrates. Proteins and lipids from tilapia waste silage have been found to have an

2
Y.S. Gaviria G et al. Heliyon 6 (2020) e05831

Table 1. Proximate and composition analysis of entrails and chemical silage.

Viscera* Chemical silage*

Moisture (%) 61,36  0,29 82,73  0,10
Crude protein (%) 10,45  0,10 50,89  0,51
Carbohydrate (%) 2,56  0,09 0,48  0,04
Crude fat (%) 85,24  0,04 39,78  0,13
Ash (%) 1,75  0,04 8,85  0,02
*
Dry matter.

appropriate nutritional profile for herbivorous and omnivorous species Table 4 shows that the values for the parameters of the red series
mainly, which can be used as a partial substitute for fishmeal (Goosen (erythrocytes, hemoglobin and hematocrits) and the white series (leu-
et al., 2014; Güllü et al., 2014), which is the most important source of kocytes and lymphocytes), obtained for the control diet are above those
protein used in animal feed (precisely due to its digestibility and protein of the silage diet. This suggests an increase in metabolism and, conse-
content (Chalamaiah et al., 2012). In this vein, this study is part of the quently, a better physiological behavior for the transport of oxygen and
current trend of evaluating the effects of using mixtures of agricultural nutrients to the tissues, thus promoting improvements in the productive
and aquaculture sources in order to reduce the environmental impact variables and a better response disposition of the immune system in
generated by fish farming and thus reuse food sources to make it sus- animals fed with the silage diet. The normal hematocrit of birds is known
tainable (Sm arason et al., 2017). Our results coincide with Kjos et al. in to vary from 35 to 55%. As mentioned above, the hematocrit can suffer
(2001) who reported similar values of protein in silage of Salmon resi- alterations with relation to the sex and age of the birds (Schmidt and dos,
dues (Salmo), which was used to feed laying hens (Kjos et al., 2001). 2007; Thrall, 2004). The white series or leukocytes are closely related to
This study was able to include a considerable fraction of viscera silage the response of the animal's defense or immune system. Heterophilia
(43.4%), which corresponds to 17.8% on a dry basis. This represents a (increased neutrophils or heterophiles), generally occurs in response to
substitution of 57.37% and 23% of soybean meal and fishmeal, respec- inflammatory or infection processes. Mild to moderate leukocytosis with
tively. Other studies have used between 15% and 40% of the silage heterophilia and lymphopenia indicates a stress response, with excess
produced, with similar physicochemical parameters with regard to pro- endogenous or exogenous glucocorticoid. This could be the case of the
tein and fat, in food for tilapia (Llanes et al., 2010). This is important to birds with slight leukocytosis in the study's silage group (Latimer and
highlight because in this case the use of silage would be maximized and Bienzle, 2000; Thrall, 2004). For example, physiological lymphocytosis
possibly reduce the costs of the production system. represents a transitory phenomenon in birds after excitement, fear or
The microbiological composition of the chemical silage of red tilapia "struggle" during handling (for example, during the blood withdrawal
viscera (Table 2), is within the parameters established by the national procedure), which coincides with what was observed in the silage group
technical directive for raw materials and inputs for animal feed of the (Latimer and Bienzle, 2000). Similar results were reported by (Cardoso
Colombian Institute of Agriculture (Directivas tecnicas de alimentos para et al., 2002), in White Leghorn birds, using a vegetable substrate in their
animales y sales mineralizadas, 1999). This shows that the chemical diet.
silage of tilapia viscera can be used as a raw material in the elaboration of This study also obtained higher than normal values for basophils,
food for Isa-Brown laying hens without causing any negative effect or possibly associated with physiological stress during the handling of birds
pathology in the species. and with stress from the laying cycle of chickens (Schmidt et al., 2009).
Two dietary treatments were used. The CT group was fed with the diet Basophilia also occurs in respiratory diseases and severe tissue injuries,
without the viscera silage, and the silage group (ENS) was fed with the associated with stress due to internal and external parasitism and is
diet of which 32% of the protein was from the viscera silage (Table 3). generally accompanied by eosinophilia (Latimer and Bienzle, 2000;
Schmidt and dos, 2007). In summary, the results show that the substi-
3.2. Hemogram and blood chemistry tution of the protein in the diet of laying hens with viscera silage did not
significantly change the blood concentration of the parameters
Table 4 shows the results of the hematological examinations with evaluated.
their respective reference values according to Schmidt et al. (2009). It is Cholesterol is an important precursor to cholesterol ester, bile acids,
important to highlight that the literature reports few reference parame- and steroid hormones and can be synthesized by various tissues in the
ters for the species which was studied (Gutierrez Castro and Corredor body, but the liver is the main organ of endogenous cholesterol synthesis
Matus, 2017). However, blood is essential for electrolyte and water (Maldonado Saavedra et al., 2012). Hypercholesterolemia can be caused
balance, for temperature control and for the functioning of the immune by diet or by liver failure (Kaneko et al., 2008). Due to the fact that
system, among other factors (Voigt, 2003). Similarly, blood values can be cholesterol is eliminated in the form of bile acids, its increase in plasma is
influenced by nutritional conditions, sex (estrogens in birds tend to associated with extrahepatic biliary obstruction, liver fibrosis, or bile
depress erythropoiesis), age, habitat, season, reproductive status, duct hyperplasia in birds (Campbell, 2004). Plasma concentrations for
trauma, management and environmental stress (Campbell, 2004). most bird species range from 100 to 250 mg/dL (Lumeij, 1997). In this

Table 2. Microbiological composition of SQ.

Result Reference
Mesophilic aerobic (ufc/ml) 5  103 10  105
2
Total coliforms (ufc/ml) 3,5  10 10  104
Fecal coliformes ABSENCE ABSENCE
Salmonella spp ABSENCE ABSENCE
Spore Clostridium ABSENCE 20  101
2
Molds and yeasts (ufc/ml) 8,2  10 10  104

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Y.S. Gaviria G et al. Heliyon 6 (2020) e05831

Table 3. Proximate composition analysis of diets.

Control Diet (%) Silage Diet (%)

Fish meal 13 10,06
Soybean meal 16,00 6,82
Corn meal 39 39,27
Rice flour 16 13
Fish entrails silage 0 17,18
Fish oil 4 0
Calcium carbonate 8 8,05
Dicalcium phosphate 2,5 4,02
Vit. min. supplement1 0,5 0,4
Lysine 0,25 0,3
Methionine 0,25 0,3
Tryptophan 0,25 0,3
Threonine 0,25 0,3
Chemical composition
Protein 20,3 20,6
Crude fiber 2,7 2,2
Carbohydrate 37,66 36,21
Crude fat 9 7,09
Metabolizable energy (kcal/kg) 3600 3500
Calcium 4 4,06
Phosphorus 0,9 1,1
1
Mineral vitamin supplement (composition per 250 g of product): vit. A - 1,400,000 IU; vit. B1 - 500 mg; vit. B12 - 300 mg; vit. B2 ¼ 500 mg; vit. B6 – 1,6 g; vit. D3 -
2,500,000 IU; vit. E - 6,000 IU; vit. K3 ¼ 1,000 mg; biotin - 30 mg; niacin -12 g; folic acid - 1 g; cobalt - 50 mg; Copper - 3,000 mg; Iron - 25 g; Iodine - 500 mg; Manganese
- 32.5 g; Selenium - 100.50 mg; Zinc - 22.49 g.

Table 4. Hemogram and blood cell count.

Unit Control Silage Reference

Leukocytes Cell x 103/μl 7,4  0,2a 11,6  0,12b 3,0–11,0
lymphocytes Cell x 103/μl 2,37  0,09a 4,87  0,11b -
Intermediate cell count Cell x 103/μl 0,07  0,01a 0,12  0,05b -
granulocytes Cell x 103/μl 4,96  0,04a 6,61  0,02a -
Lymphocytes % 32  0,7a 42  0,5b 20–65
Monocytes % 1  0,2a 1  0,08a 0–5
Heterophils % 49  0,8a 47  0,3a 30–75
Eosinophils % 6  0,3a 2  0,7b 0–5
Basophiles % 12  0,07a 8  0,04b 0–5
Bands % 0a 0a -
Erythrocytes Cell x 106/μl 3,97  0,03a 4,3  0,06a 2,5–4,5
Hemoglobin g/dl 9,7  0,2a 10,6  0,2a 11,0–19,0
Hematocrit % 29  0,37a 32  0,57a 23,0–55.0
Platelets Cell x 103/μl 44,3  0,83a 49  0,36b -
Plasmatic proteins g/dl 3,03  0,06a 3,8  0,07a 3,0–4,9
a-b
represent a significant difference p < 0,05 for the groups; n ¼ 3 in each group.

study, the cholesterol levels of the silage group were 71.33 mg/dL vs 114 et al. (2007), who observed a decrease in cholesterol and serum tri-
mg/dL of the control group, and the triglycerides were 63 mg/dL vs 170 acylglycerides in laying hens, when using 0.04% Rhodobacter capsules
mg/dL for each, respectively. In this regard, the levels of cholesterol and (bacteria, probiotic). Nogueira et al. (2003) also found similar values,
triglycerides in silage diet were decreased by 37.4% and 62.9% when supplementing diets with shark cartilage (rich in polyunsaturated
compared with those in control group. These differences in cholesterol fatty acids). The reverse effect is seen in diets with high percentages of
and triglycerides can be attributed to the percentage of crude fat in the saturated fatty acids, since they stimulate the hepatic production of very
diets. low-density lipoproteins (LMBD), by stimulating the expression of
Similarly, omega 3 and omega 6 lipids increase high-density lipo- apolipoprotein B-100 (Bennett et al., 1995).
proteins (LAD) and decrease atherogenic conditions and endothelial Likewise, high levels of saturated fatty acids affect the lipid profile of
dysfunction in the blood vessels of birds (Palou et al., 2005), which im- broilers with compatible signs of ascites syndrome, where a depression of
proves blood supply and consequently the quality of the egg as a final the energy metabolism was observed due to decreased energy sources,
product for the benefit of the end consumer (Buitrago et al., 2005). since the animal gradually loses its appetite. The loss of appetite is
Similar results to those of the present work have been reported by Salma generated by the pressure caused by abdominal edema and may lead to

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Y.S. Gaviria G et al. Heliyon 6 (2020) e05831

Table 5. Blood chemistry.

Unit Control Silage Reference

ALT U/L 1,3  0,1a 4,8  0,2b
Alkaline phosphatase U/L 44,69  0,17a 39,25  0,13a 10–106
a
Total protein g/dL 3,56  0,27 4,84  0,21a 3–4,9
Cholesterol mg/dL 114  0,8a 71,33  1,7b 129–279
Triglycerides mg/dL 170  1a 63  1,2b 79–517
a-b
represent a significant difference p < 0,05 for the groups; n ¼ 3 in each group.

100

90

80

70

60
LAYING (%)

50

40

30
Control
20
Silage
10

0
0 1 2 3 4 5 6 7 8 9 10 11 12 13
TIME (Weeks)

Figure 1. Laying percentage as a function of time during feeding of Isa-Brown line laying hens (Gallus gallus domesticus).

80

70

60

50
WEIGHT (gr)

40

30
Control
20
Silage
10

0
1 2 3 4 5 6 7 8 9 10 11 12 13
TIME (Weeks)

Figure 2. Egg weight as a function of time during feeding of Isa-Brown line laying hens (Gallus gallus domesticus).

decreased glucose, cholesterol and triglyceride levels (Buitrago et al., Plasma (Table 4) and total (Table 5) proteins are within the reference
2005). ranges, which indicates an adequate performance of both diets (control
Table 5 shows that the triglyceride and cholesterol values are lower in and test). Total plasma proteins in birds vary from 3.0 to 4.9 g/dL, of
the blood of animals fed with silage feed, compared to the control, which albumin comprises 40–50% (Kaneko et al., 2008). In general, the
possibly due to the presence of monounsaturated fatty acids in the viscera main factors that affect total protein concentrations in birds are: age,
of fish (red tilapia), according to Clarke et al. (2002). This phenomenon seasonality, handling conditions, diseases and egg production (by albu-
as yet to be well documented in birds. However, we know that foods that min and globulins) (Lumeij, 1997). In this vein, the results show proper
are rich in monounsaturated fat promote cardiovascular health in handling according to the age and production stage of the eggs, in
humans, because it causes a reduction in postprandial triglycerides with addition to the absence of protein deficiency diseases in the birds which
respect to the intake of saturated fat (Palou et al., 2005). were studied. Furthermore, animal sources of protein are known to have
Similarly, Table 5 shows that the levels of the enzyme alanine a higher quality and biological value than plant protein (Rosenfeld et al.,
aminotransferase (ALT) for both the control and silage group were below 1997).
the reference levels. The ALT enzyme is found both in the hepatocyte
cytosol and in muscle and other tissues in birds. ALT activity in most bird 3.3. Productive variables
species varies from 19 to 50 U/L (Campbell, 2004; Lumeij, 1997), when
this enzyme is above this range, it indicates possible liver damage, Figure 1 shows that there were no statistically significant differences
especially in its functionality. However, no possible negative effects are (P > 0.05) in the percentage of egg-laying between the group fed with the
reported for low ALT levels (Sanchez, 2009). control diet and the diet made with silage. The data shows that the

5
Y.S. Gaviria G et al. Heliyon 6 (2020) e05831

2400.00

2200.00

2000.00

WEIGHT (gr
1800.00

1600.00

1400.00

1200.00

1000.00
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14
TIME (Weeks)

Figure 3. Weight gain over time of Isa-Brown line laying hens (Gallus gallus domesticus) during their feeding.

stability in the laying percentage (at 85%) was reached in week 5 for both to be used as a protein raw material in the feeding of different species of
diets and was maintained during the following weeks of study. This birds. For this reason, this kind of silage can be used as an alternative
means that the quantity and quality of degreased silage in laying hens' protein raw material in the elaboration of diets for laying hens, thus
feed did not have an adverse effect on the laying percentage of the birds, substituting conventional raw materials such as fishmeal and soybean
which shows that the nutritional requirements were adequately adjusted meal, without modifying the productive variables, percentage of laying,
for the expected level of laying. Similar results were found by Kjos et al. or weight gain. Likewise, the diets that include silage have positive ef-
(2001), however, this study only used 5% silage in the feed. Additionally fects on the health and nutrition of the birds within the established pa-
(Al-Marzooqi et al., 2010), these authors showed that replacing soybean rameters, and do not show any negative alterations in blood chemistry or
meal with fish silage protein improved the growth performance of hemograms (from the red and white series). The inclusion of acidic silage
broilers. In general, the amino acid content and protein quality of animal from red tilapia viscera in the diet of laying hens significantly decreased
sources tend to be of better biological quality and consequently obtain the percentage of blood cholesterol and triglycerides in the birds.
better responses in weight and production compared to diets from plant
sources (Rosenfeld et al., 1997). Declarations
Figure 2 shows that the average weight of the eggs increased gradu-
ally over time, until constant values were maintained between the sixth Author contribution statement
and seventh week, without showing statistically significant differences
between the diets (P > 0.05). Padhi et al. (2013) reported a similar Yhoan S. Gaviria G.: Performed the experiments; Analyzed and
behavior in the increase of egg weight depending on the age of the birds, interpreted the data; Wrote the paper.
concluding that the egg increases in weight until week 52, after which it Luis F. Londo~ no F.: Conceived and designed the experiments;
maintains its uniform weight (Padhi et al., 2013). In this study both diets Analyzed and interpreted the data; Wrote the paper.
maintained a similar trend until week 12 of laying, after which the silage Jose E. Zapata M.: Conceived and designed the experiments; Analyzed
diet achieved a significant increase in egg weight (P < 0.05), possibly due and interpreted the data; Contributed reagents, materials, analysis tools
to the fact that the diet with silage had higher levels of plasma (Table 4) or data; Wrote the paper.
and total (Table 5) proteins. These proteins are the primordial precursors
of the yolk (vitellogenin and lipoproteins), are synthesized in the liver Funding statement
and transported to the ovaries where they are incorporated into the
oocyte in the ovary (Campbell, 2004; Lumeij, 1997; Rosenfeld et al., This work was supported by Comite para el Desarrollo de la Inves-
1997). tigaci
on en la Universidad de Antioquia (CODI) (Sustainability program)
Egg weights were consistent those obtained by Batalha et al. (2018), and COLCIENCIAS (111574558746).
who used a lower concentration of silage in the diet. This may suggest
that using higher concentrations of silage does not adversely affect the
Data availability statement
weight of eggs over time, and could actually could favor it.
On the other hand, the weight gains of the birds in both diets did not
Data included in article/supplementary material/referenced in
show statistically significant differences during the weeks of the study (P
article.
> 0.05) (Figure 3). Abelti (2018) obtained similar results by feeding
broilers by adding different levels of Barbus paludinosus silage (Abelti,
2018) in the feed. Additionally (Al-Marzooqi et al., 2010), these authors Declaration of interests statement
showed that replacing soybean meal with fish silage protein improved
the growth performance of broilers. In general, the amino acid content The authors declare no conflict of interest.
and protein quality of animal sources tend to be of better biological
quality and consequently obtain better responses in weight and produc- Additional information
tion compared to diets from plant sources (Rosenfeld et al., 1997).
No additional information is available for this paper.
4. Conclusions
References
The chemical silage of red tilapia viscera has the appropriate micro-
biological conditions in accordance with Colombian national regulations Abelti, A.L., 2018. Evaluation of small barbus silage through inclusion into commercially
formulated poultry feed. Int. J. Poult. Sci. 2 (1), 1–7.

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