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Article

Pilot Study of Diet Supplemented with Sold-Out Substrate of Pleurotus ostreatus in the Feeding of Backyard Broilers

by
Omar Romero-Arenas
1,
Maribel Martínez Carrera
1,
Gerardo Landeta-Cortés
2,
Victor Rodríguez-Hernández
3,
Nemesio Villa-Ruano
4 and
Antonio Rivera
5,*
1
Centro de Agroecología, Instituto de Ciencias, Benemérita Universidad Autónoma de Puebla, Edificio VAL 1, Km 1,7 Carretera a San Baltazar Tetela, San Pedro Zacachimalpa, Puebla 72960, Mexico
2
Centro Universitario de Vinculación, Benemérita Universidad Autónoma de Puebla, Puebla 72570, Mexico
3
Colegio de Posgraduados, Campus Puebla-México, Puebla 72860, Mexico
4
CONACyT-Centro Universitario de Vinculación y Transferencia de Tecnología-DITCo, Benemérita Universidad Autónoma de Puebla, Cd. Universitaria, Puebla 72570, Mexico
5
Centro de Investigaciones en Ciencias Microbiológicas, Instituto de Ciencias, Benemérita Universidad Autónoma de Puebla, Ciudad Universitaria, Puebla 72570, Mexico
*
Author to whom correspondence should be addressed.
Appl. Sci. 2023, 13(13), 7428; https://doi.org/10.3390/app13137428
Submission received: 4 May 2023 / Revised: 12 June 2023 / Accepted: 15 June 2023 / Published: 22 June 2023
(This article belongs to the Special Issue Livestock and Poultry Production：Technologies and Prospects)
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Abstract

:
There are several by-products obtained in agricultural systems that can contribute to small-scale production systems within backyards, in this sense, the production of oyster mushrooms (Pleurotus ostreatus) has been integrated into the rural family economy in Mexico. After harvesting the fruiting bodies, the remainder is known as sold-out substrate, this by-product has been destined for other agricultural uses such as: medium for seedlings, vermiculture or fertilizer; however, there are studies where it has been used in the diet of bovine cattle. In this study, the effects of sold-out substrate (corn stubble) of P. ostreatus in the diet of broilers with different percentages of protein, on feed conversion ratio, carcass yield, and broiler meat quality were evaluated. A total of 120 broilers Ross 308 were randomly assigned in 12 pens with concrete floor and curly shaving with an area of 3 m2. During the initial period (1 to 28 days), all broilers were fed commercial feed® mixture. On day 29, the broilers were weighed and divided into four treatment groups and fed diets with different percentages of protein until the end of the experiment. Feed conversion ratio was significantly higher in treatment control (TC) compared to the other treatments; however, the performance parameters were not affected by the diet supplemented with sold-out substrate, likewise, the color and quality of the meat were in a normal range (48 < L* < 51) and with a good amount of crude protein. Sold-out substrate of P. ostreatus from corn stubble presented a high mineral content of Mg, Na, K, Fe, Cu, and Mn suitable to be considered in diets for feeding backyard broilers.

1. Introduction

Poultry (Gallus gallus domesticus) production on a small scale is a common practice in rural households around the world. Various studies have shown that poultry farming continues to play an important socioeconomic role in rural development [1,2,3].
In Mexico, poultry production systems are developed under different agroecological and technological contexts with differences in management and production objectives; in general, they are classified into three schemes: (a) technical, (b) semi-technical, and (c) traditional or backyard; the latter channels its production to family self-sufficiency [4]. Backyard poultry farming is the most traditional and widespread activity in agricultural areas; it has been carried out since colonial times [5], it provides 85% of food security in rural productive units of the country, and it delivers high-quality food such as eggs and meat to meet essential family needs [6]. However, backyard poultry production systems are maintained without adequate disease prevention or control strategies, inadequate management practices, and poor feeding [7].
The supply of food for backyard poultry farming in Mexico and Latin American countries consists mainly of providing corn in its different forms and transformations, plants, and waste from the family food consumption; this type of diet does not satisfy the basic nutritional requirements of birds [8]. Feeding backyard broilers requires simple carbohydrates, fat, and protein for energy; proteins are extremely important, since their function is to provide necessary amino acids for muscle maintenance and development [9]. Therefore, all efforts are aimed at maintaining the quality of diet consumption and reducing the cost of feed to ensure maximum conversion of nutrients with high biological value [10].
To achieve nutritional requirements in backyard broilers, family production units have sought ways to meet this nutritional demand through commercial feed® or diets with high protein content, therefore, production costs increase and decrease their earnings [11]. Commercial feeding represents the highest cost of poultry production, oscillating between 65 and 70% of total costs [10,12], for which it is necessary to look for alternatives to reduce input costs without affecting the nutritional parameters in backyard broilers, likewise, they must be within the same locality and be easily obtained [11,13,14].
In a study conducted by Giannenas et al. [15] where Agaricus bisporus flour was incorporated into the diet of broilers, positive results in promoting growth and antioxidant protective activity in tissues were obtained, in addition to positively affecting the intestinal health of birds. The sold-out substrate of P. ostreatus has been used in the feeding of polygastric and monogastric animals [16,17]. In this sense, various studies address that the sold-out substrate or remaining substrate of P. ostreatus constitutes a source of predigested proteins, minerals, and fibers, with variations in the nutritional content of substrate, since it depends on the agricultural residue that is used to produce oyster mushrooms [18].
Currently, edible mushrooms of genus Pleurotus are cultivated in a wide range of altitudes and have the capacity to grow in different plant residues, representing an alternative with biological, economic, and social viability for the rural population of Mexico [19]. In addition, the sold-out substrate of P. ostreatus is easy to collect and handle; it also presents a high level of N, K, P, Ca, and trace elements, particularly iron and silicon [18] necessary for the growth and development of backyard broilers [10]. Therefore, the main objective of this research was focused on evaluating the effect of sold-out substrate (corn stubble) from the production of oyster mushrooms in the diet of broilers, incorporating different percentages of protein. Corn stubble is an agricultural by-product derived from the crop harvest, consisting mainly of leaves and dry stems.

2. Materials and Methods

2.1. Characterization of Sold-Out Substrate of P. ostreatus

2.1.1. Proximal Chemical Analysis

Five hundred grams of sold-out substrate of P. ostreatus from corn stubble aged 90 days was dried in a Memmert Beschickung (Memmert GmbH, modelo 100-800, Schwabach, Alemania) oven at 60 °C for 24 h until reaching constant weight; 90 days was selected since it is the time that the production cycle of the oyster mushroom culture lasts. The material was subjected to a mechanical grinding process in a chopper mill (TRAPP TRF 300 G, Pulvex, Mexico City, Mexico) for 30 min. Subsequently, a hammer mill (WERKE M-20, IKA, LEYVITEC Laboratorios S.A. de C.V. Mexico City, Mexico) was used to obtain a particle size of 0.5 cm.
Proximal chemical analysis was performed following procedures recommended by regulations of the Association of Official Analytical Chemists [20,21]. Crude protein (CP) was determined using the micro-Kjeldahl method with a conversion factor of 6.28 [20,21]. Lipid determination was carried out using the Goldfish AOAC technique no. 95-402 [20,21]. Ash content was determined by constant weight ashing at 550 °C in a muffle furnace [20,21], while crude fiber was evaluated by the Van Soest method [22]. Dry matter (DM) was maintained for 36 h at a temperature of 55 °C; once constant weight was reached it was calculated with the following formula: DM = [final weight of the sample/initial weight of the sample] × 100 [23]. Carbohydrate content present in samples was calculated using total carbohydrate (C) method [24]. This value was calculated according to the following expression (Equation (1)):
C = 100% − (moisture% + crude protein% + fat% + ash%)
where C = carbohydrates in 100 g of dry matter.
The energetic value was calculated using the specific factors according to the following expression [24] (Equation (2)):
Energy = (C × 4) + (CP × 4) + (F × 9)
where Energy = Kcal in 100 g of dry matter; C = carbohydrates (excluding dietary fiber); CP = crude protein, and F = fat.

2.1.2. Mineral Element Analysis

Five hundred grams of sold-out substrate of P. ostreatus from corn stubble aged 90 days was dried at a temperature of 60 °C for 36 h. Subsequently, it was crushed until obtaining a particle size of 0.2 µm. Finally, the dried and pulverized samples weighing 0.5 g were placed in eight tetrafluoromethoxy (TFM) vessels, together with 9 mL of 65% (v/v) HNO3 and 3 mL of HCl (1:1). The first microwave digestion was carried out for 5 min with a power of 700 W at a temperature of 180 °C. The second digestion was performed at 180 °C with a power of 500 W for 10 min, which was verified with a temperature probe in a control vessel. Once the digestion was complete, the vessels were allowed to cool, and the resulting solution was filtered to remove possible residual particles.
Ultimately, each supernatant was transferred separately to 50 mL capacity flasks and was measured to 25 mL with deionized water. The resulting solutions were kept in plastic bottles that were previously washed with deionized water and stored at 4 °C. Minerals were identified and quantified using a multielement ICP standard solution (CertiPUR, Merck, KGaA, Darmstadt, Germany) containing 23 standard elements using a Varian Axial 720 Inductively Coupled Plasma Optical Emission Spectrophotometer (ICP-OES, Varian, Palo Alto, CA, USA) to determine mineral content (Ca, Mg, Na, K, P, Fe, Cu, Mn, and Zn). Data and standard curves were processed using ICP-Expert™ II (Varian, Palo Alto, CA, USA) [25].

2.2. Location

The study was carried out during the months of September to November 2021 at the facilities of the Family Farm of the Ejidal Commissariat of the municipality of Tlacotepec de Benito Juarez, Puebla-Mexico (18°41′04″ N 97°39′12″ W), at an altitude of 2004 m.a.s.l. Climatic conditions of the region are characterized by an average temperature of 17 °C, annual rainfall between 200 and 400 mm, and a temperate semi-dry climate with summer rainfall [26].

2.3. Experimental Design and Diets

Care and management of birds were agreed upon and approved by the Bioethics Committee of Facultad de Medicina Veterinaria y Zootecnia, and the Posgrado en Manejo Sustentable de Agroecosistemas of Benemérita Universidad Autónoma de Puebla (FACMEDVET/CI2021-2022).
The sold-out substrate of P. ostreatus (from corn stubble) was subjected to particle reduction with the help of a manual mill, until reaching an approximate size of 0.5 mm, which was incorporated into the different experimental diets in the growth and completion stage.
The dietary experiment (pilot study) was conducted on 120 Ross 308 broilers from a commercial hatchery; 10-day-old male chicks were vaccinated with Tabic VH® against Newcastle disease (1 drop per eye per bird) and TAbic H-120 Var® against avian influenza (0.5 mL/bird) [27]. Commercial feed® was supplied in the initiation stage, from day 1 to 28 days of age, reaching an average weight per bird of 0.48 ± 0.01 g. The commercial feed® contains 20.16% protein that provides 3275 (kcal/kg) of metabolizable energy, necessary for the nutritional contribution of broilers (Table 1).
For the growth and finishing stage, the broilers were housed in groups of 10 birds in 12 pens with a concrete floor and curly shaving bedding with an area of 3 m2 each, with a density of 3.32 birds/m2, equipped with an automatic bell-type drinker and tubular feeder [28]. The distribution of the experimental flock was randomized following a completely randomized design in 4 groups of 10 broilers, with three repetitions per treatment.
The diets were divided into a control treatment (n = 30) based on commercial feed®; treatment T1 (n = 30) based on sold-out substrate, wheat germ, and corn tortilla; treatment T2 (n = 30) based on sold-out substrate and wheat germ, and T3 treatment (n = 30) based on wheat germ and corn tortilla (Table 2), according to the calculations made through the Pearson square; this being a practical method when mixing two or more foods that have different concentrations of protein [29]. Metabolic energy was calculated for feed components based on the results of chemical analyses, prior to formulating dietary recipes, based on poultry recommendation guidelines for birds (Table 2).
From four weeks onwards, the broilers began to be fed with different diets (Figure 1) and free access to water; every 24 h, the pens were cleaned of food remains until reaching 70 days of the experiment. Every 15 days, the sawdust bed was changed and new curly shaving bedding was placed.

2.4. Feed Conversion Ratio (FCR)

To determine the productive behavior of each experimental diet in comparison with the treatment control, the feed intake and weight gain of each bird per treatment were evaluated. The feed conversion ratio was calculated by adding the total feed consumed between the weights of the broilers (FCR = TFC/WB) [30]; average water consumption per treatment was also recorded.

2.5. Determination of Anemia and Cholesterol

For the determination of anemia and cholesterol, three broilers were randomly taken per treatment and were fasted for 12 h; later, 1 mL of blood was extracted from the branchial vein of each bird. The complete blood count was determined using the methodology proposed by Mitchell and Johns [31], analyzing parameters of hematocrit, hemoglobin, and white blood cells. Regarding blood chemistry, cholesterol levels were determined using the dry chemistry method with an ion-selective electrode as established by Franco et al. [32].

2.6. Carcass Weight

Upon completing 10 weeks of feeding in different groups, experimental work was completed, and six broilers were sacrificed per treatment randomly chosen, which were subjected to 6 h of fasting. Birds were weighed before and after slaughter, recording the different variables: live weight (LW); slaughter weight (SW); blood weight (BW); carcass weight with viscera (CWV); carcass weight without viscera (CWwV); feather weight (FW); visceral fat weight (VFW); breast muscle weight (BMW); thigh muscle weight (TMW); wing weight (WW); head and neck weight (HNW); gizzard weight (GW), and liver weight (LW) [33], in order to determine if feeding had an effect on productive parameters.

2.7. Proximal Chemical Analysis of Broiler Meat

The pieces of breast (BMW) and thigh muscle weight (TMW) without skin from each carcass treatment were used to determine the percentage of protein according to the method indicated in the Association of Official Analytical Chemists in triplicate [20]. Crude protein content was determined by the Kjeldahl method with a conversion factor of 6.25. Crude fat content was determined by the Soxhlet method with diethyl ether as solvent. Ash content was determined by a constant weight at 550 °C in a muffle furnace. Finally, the percentage of humidity was determined by drying the sample at a temperature of 105 °C at a constant weight.

2.8. Determination of pH and Color

A penetrating electrode was used at three different points of the broiler muscle, using a portable pH meter (Mod SG2, Mettler Toledo AG, Schwerzenbach, Switzerland) for meat, to perform the analysis of this variable. The color of pectoral muscle surface was measured in triplicate using a colorimeter (Minolta Chroma Meter CR 400) programmed with the CIE-Lab* system and expressed in L* (lightness), a* (redness), and b* (yellowness) values [34].

2.9. Statistical Analysis of the Results

In the present investigation, data were statistically analyzed as a completely random design according to Equation (3):
yij = μ + τi + εij
where yij = value of the observation; μ = average of the population; τi = treatment effect, and εij = experimental error.
Data obtained were processed with analysis of variance (ANOVA), and Tukey’s multiple comparison test (p < 0.05) was applied to determine the differences between the treatments using the statistical package SPSS Statistics version 17 (Statistical Package for the Social Sciences).

3. Results and Discussion

3.1. Analysis of Sold-Out Substrate of P. ostreatus

The crude protein, moisture, ether extract or fat, organic matter, crude fiber, ash, and mineral matter contents of sold-out substrate of P. ostreatus (from corn stubble) are shown in Table 3.
Romero et al. [35] mentioned that sold-out corn stubble substrate presents 4.9% crude protein and 0.37% ethereal extract, results similar to those obtained in the present investigation. The value obtained in organic matter was 63%, likewise, Bermúdez et al. [36] reported a value of 75.91% in coffee pulp. The organic matter of sold-out substrate achieves a reduction due to the losses of H2O and CO2 during the process of mycelial metabolism and fruiting [37].
Percentage of crude fiber obtained was 16.4%. This differs from the metabolism of the mycelium, which fulfills the function of degrading lignocellulosic matter, transforming it into simpler compounds, in addition to obtaining a higher percentage at the end of the fungus harvest, as demonstrated by Bermúdez et al. [36], with the pleurotin of coffee.
Metabolizable energy obtained for each kg of sold-out substrate was 2836.1 (kcal/kg); in this sense, Ferreira et al. [38] showed that dietary energy close to 3000 kcal/kg did not affect body weight in broilers. When the diet contains low energy values, broilers consume more feed and feed conversion ratio will be less efficient; however, the cost of this energy must be considered as a higher energy content in the diet does not always mean a higher economic benefit [39].
Data collected by Santiago [40] maintains values of Fe = 50, Cu = 10, Mn = 65, and Zn = 60 mg/Kg in broiler diets; these values turn out to be similar in comparison to the results obtained in this work (Fe, Cu, Mn, and Zn with 765, 3, 113.5, and 29.5 mg/kg, respectively). Mineral elements, including Cu, Fe, and Mn, play an important role in poultry production [41].

3.2. Production Performance

The performance parameters of broilers fed with different percentages of protein inclusion using the sold-out substrate of P. ostreatus are presented in Table 4.
Initial weight of the broilers was similar in all replicates of each treatment group. However, a greater total weight gain was observed in the treatment control with 2.33 ± 0.34 kg, with respect to the T1 and T2 treatments, while the T3 treatment obtained the second-best weight gain, presenting significant differences (p < 0.019).
In this study, food and water intake showed significant differences between the diets of each group (p < 0.003). The best feed conversion ratio compared to the other treatments was obtained by the treatment control, with a reported value of 1.53 ± 0.10. Treatment T1 obtained the highest value, while the T2 and T3 treatments presented a feed conversion ratio with values of 1.66 ± 0.15 and 1.57 ± 0.94, respectively, however, no significant differences were observed.
Campo et al. [12] reported higher feed conversion ratio values (2.2 and 1.97) with commercial feed® with the inclusion of 10 and 20% chontaduro husk meal, which were enriched with P. ostreatus. That is why soluble polysaccharides without starch create a high viscosity in digestion and improve weight gain, in addition to reducing FCR in broilers [42]. It has been shown that there are factors that influence the feed conversion ratio of broilers, such as the temperature that affects feed consumption, causing stress; however, even though consumption is normal, the speed of passage increases and decreases the digestion of nutrients, and therefore, the feed conversion ratio increases [43].
Another important factor is the size and shape of the feed particles. In our study, the food from the control treatment was in the form of granules, and the other treatments were in prepared flour. Industrial diets are made in the form of granules, with small particle size, in order not to generate waste at the time of ingestion [44], since they allow a better apprehension of food, which generates less waste and, at the same time, there is greater ingestion of nutrients, generating a positive response in the feed conversion ratio [45]. In this sense, it is suggested that feed presentation has a very significant impact on the growth of broilers [46]; however, it has also been observed that excessive commercial feed® intake may not improve performance growth rate, but could reduce digestibility, nutrient absorption, and increase the rate at which food materials pass through the gastrointestinal tract [47,48].
Blood biomarkers are important indicators of the nutritional status of broilers. The analysis of blood parameters is considered an important tool to investigate the health, physiological, pathological, and nutritional status of animals [49]. Values obtained for hematocrit parameters in the present study (Table 4) were within the range reported in the literature [50,51,52,53,54,55]. Therefore, it suggests that the dietary inclusion of sold-out substrate is safe for consumption by broilers.
Cholesterol values (2.80–3.43 mmol/L) obtained are within the normal range (7.74–17.82 mmol/L) reported by the Clinical Diagnostic Division, as mentioned by Daudu et al. [56]. However, the blood cholesterol level in the broilers of treatment control was higher than that of the other treatments (3.43 mmol/L), while cholesterol levels in sold-out substrate treatments were significantly reduced.
Inclusion of sold-out substrate in the feed helps to reduce the cholesterol content in the blood. A study conducted by Kim et al. [57] found low concentrations of cholesterol in the group fed by fermented rice bran than in the control group for laying hens. Wang et al. [58] reported that fermented food helps to inhibit the activity of 3-hydroxy-3-methyl-glutaryl-CoA reductase, which reduces cholesterol biosynthesis. These results are similar to those obtained in the present investigation, where sold-out substrate comes from a solid fermentation carried out in the production of fruiting bodies of P. ostreatus [59].

3.3. Analysis of Carcass Traits

The weights of slaughtered broilers do not present significant differences among themselves (Table 5); however, it was observed that in treatment control, broilers were heavier with an average of 2774 g; this may be due to the consumption of less protein that contains the wheat germ, sold-out substrate, and corn tortilla. These results were like those reported by Bello-Pérez et al. [60] in broilers fed traditional and commercial corn tortillas with protein content of 7.82 and 7.73%, respectively. Similarly, no significant differences were observed in carcass weight with viscera (CWV), carcass weight without viscera (CWwV), breast muscle weight (BMW), thigh muscle weight (TMW), head and neck weight (HNW), and gizzard weight (GW). This may be due to the inclusion of fungal mycelium which positively affects body weight and the balance of microflora in the broiler intestinal tract, leading to more effective utilization of dietary nutrients [61].
There are significant differences in the relationship between organ weight and body weight (Table 5). The treatments T1, T2, and T3 have a lower weight in blood compared to the treatment control, likewise, the weight of the feather found in the different treatments presents significant differences (p = 0.09). Additionally, they presented significant differences in the weight of visceral fat; the treatments T1, T2, and T3 have a lower weight of visceral fat compared to the treatment control, this may be due to consumption of the amount of crude fiber containing wheat germ, sold-out substrate, and corn tortilla. Crude fiber is slowly soluble in the digestion process; when examining the bird intestines, a large amount of food was found, and this was corroborated by the volume of their feces and their rigidity during the time the study lasted. In addition, within the treatment control, it was observed that their behavior was more peaceful compared to the T1 and T2 treatment that presented hyperactivity; in addition, they obtained a lower gizzard weight to body weight ratio.
Dietary inclusion of AGPI-rich oils has been reported to enhance oxidation and decrease FA synthesis, with a resultant reduction in abdominal fat accumulation in broilers [62,63]. In this sense, Alsanad et al. [64] showed the profile of fatty acids presented by the sold-out substrate of P. ostreatus in four different treatments, where it could be observed that it contained 41.2% linoleic acid, 9.7% oleic acid, 9.6% stearic acid, 3.6% linolenic acid, and 0.9% arachidic acid. In this sense, it has been reported that the supplementation of mushrooms and probiotics in poultry feed shows beneficial and synergistic effects on the immune response, performance, and serum lipids in broilers [65]. In the present study, it was found that broilers fed with commercial treatment developed more abdominal fat, had less physical activity, and consumed more water at the end of the experiment.

3.4. Meat Quality

In general, there were significant differences (p < 0.005) in physical parameters of the breast and thigh meat of broilers fed with the sold-out substrate of P. ostreatus (from corn stubble) with different percentages of protein compared to those fed with the treatment control diet. In this study, the pH values of breast muscle (BM) and thigh muscle weight (TMW) are within the ranges of normal pH limits [66].
The color of any meat product is one of the most used indicators to determine freshness and healthiness of the meat, obtaining a good initial image of the product [67]. In the present investigation, sold-out substrate with different percentages of protein presented a significant difference (p < 0.05) in color properties of breast muscle (BM) and thigh muscle weight (TMW) meat at 24 h post-mortem between diets of the broilers (Figure 2).
Treatment T2 obtained the greatest lightness (L*), redness (a*), and yellowness (b*) unlike the other treatments, which did not present a significant difference with treatment control. In a study conducted by Stadig et al. [68], the authors found that broiler meat color characteristics ranged from 53.9 to 55.3 for lightness, 5.7 to 6.3 for redness, and 13.4 to 14.7 for yellowness, which were results higher than those reported in present investigation.
Based on the luminosity values proposed by Qiao et al. [69], meat is classified as lightest (pale, L* > 53), normal (48 < L* < 51), and dark (L* < 46), based on these values, and according to the results of the present investigation, the values of L*, a*, and b* for all the treatments were in a normal range (48 < L* < 51), which indicates that the breast muscle meat (BM) and the weight thigh muscle weight (TMW) are considered light, soft, and non-exudative [70].
Inclusion of sold-out substrate with different percentages of protein in the broiler diet influenced the moisture%, CP%, EE%, and ash content of breast muscle (BM) and thigh muscle weight (TMW) meat compared to treatment control. In a study conducted by Altmann et al. [71], the authors reported 73.88–74.29% muscle moisture; likewise, Schiavone et al. [72] obtained 75.24–76.14% moisture in broilers, results similar to those reported in the present investigation.
Crude protein (PC%) of breast muscle (BM) and thigh muscle weight (TMW) meat in the three treatments with respect to the treatment control showed significant differences (p < 0.05). Control treatment has the highest crude protein content (PC%), which is directly proportional to the amount of crude protein in the commercial feed®. Protein-rich meats are considered to exhibit good nutritional quality with a proportion of essential amino [73]. According to data collected by Bohrer [74], breast muscle (BM) can contain 22.5% of crude protein, while the thigh muscle weight (TMW) can have a value of 16.2%. Kirk et al. [75] mentioned that the protein of breast muscle (BM) and thigh muscle weight (TMW) can reach values of 23.4, 20.4, and 19.9%, respectively, which are values lower than those found in this research work (Figure 3).
The mean contents of ethereal extract (EE = 5.35%) in the present experiment were in good agreement with Schiavone et al. [72]. The highest ethereal extract content obtained in this work was presented by treatment control and T3 treatment, which do not show significant differences between them; this was confirmed by finding accumulations of fat in the breast muscle (BM) and thigh muscle weight (TMW). Treatments T1 and T2 show a lower percentage of ethereal extract in breast muscle (BM) and thigh muscle weight (TMW), coinciding with that reported by Altmann et al. [71] where they reported a considerably higher fat content of broiler meat (3.40–3.41%) with commercial feed®.
Ash content in any feed acts as a determining factor for the availability of minerals and energy metabolizable from the diet. The result of ash content shows that the T2 treatment registered higher values (1.88%) presenting significant differences (p < 0.05) with respect to the other treatments. The lowest ash content occurred in the T3 treatment, and this could indicate a lower availability of minerals and energy in food [76] available for broilers.
Okuskhanova et al. [77] investigated maral meat which showed lower protein content (19.4%), higher fat content (1.4%), and lower ash content (0.7%) compared to the results obtained in the present investigation. The high protein and ash content, in addition to the low-fat content presented in the T2 treatment, could be considered a unique nutritional characteristic with good potential for feeding backyard broilers.
The values registered for the four treatments are within the range of ash present in broiler meat (0.7–1.3%). In a study by Daniel [78], he recorded the mean value of 1.23 ± 0.130% in leg muscle ash. High levels of ash imply a lower concentration of organic matter; the average of sold-out substrate is 63 g for organic matter, results like those mentioned by Rude and Rankins [79]. Amounts less than 80% organic matter indicate a lower energy and protein content, which are the most critical and expensive nutrients in animal feed. However, feeds with 1.5% ash contain enough minerals to be considered at the time of formulation of diets for feeding broilers, with substantial savings [77].

4. Conclusions

The sold-out substrate of P. ostreatus from corn stubble presented a high mineral content of Mg, Na, K, Fe, Cu, Mn, and Zn, except for Ca and P, which must be considered for the growth and development of backyard broilers. In addition, due to its high Fe content, the treatments T1 (wheat germ, corn tortilla, and sold-out substrate) and T2 (wheat germ and sold-out substrate) obtained a higher percentage of hematocrits compared to the control treatment (commercial feed®).
The amount of lipids (cholesterol, ethereal extract, visceral fat) in the T1 and T2 treatments were low compared to the treatment control, possibly due to the increase in fiber contained in the diets with sold-out substrate. Even though the feed conversion ratio was high in treatments T1 and T2, feed and water consumption were lower compared to TC.
At week 10, no experimental treatment from the pilot study reached the weight obtained by the control treatment; however, it is necessary to remember that, in general, a protein bank is kept in the backyard for more than three months and the sacrifice is performed at the right time for the producer.
Further studies are recommended in which the complementation of ratios for broiler feeding can be determined in greater detail so that these results can become precedents and consulted for a series of future investigations.

Author Contributions

Conceptualization, M.M.C. and O.R.-A.; methodology, M.M.C. and V.R.-H.; software, N.V.-R.; validation, O.R.-A., M.M.C. and V.R.-H.; formal analysis, A.R., N.V.-R. and O.R.-A.; investigation, V.R.-H. and O.R.-A.; resources, A.R., M.M.C. and O.R.-A.; data curation, G.L.-C.; writing—original draft preparation, G.L.-C. and O.R.-A.; writing—review and editing, O.R.-A.; visualization, A.R.; supervision, O.R.-A.; project administration, A.R.; funding acquisition, M.M.C. and O.R.-A. All authors have read and agreed to the published version of the manuscript.

Funding

This research was supported by the PRODEP 2021 program of the Secretaría de Educación Pública de México (SEP); number C.A-348, the Consejo Nacional de Ciencia y Tecnología (CONACyT), number 217470628, and Benémerita Universidad Autónoma de Puebla, number 100420500.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Informed consent was obtained from all subjects involved in the study.

Acknowledgments

The authors thank the Family Farm of the Ejidal Police Station of the municipality of Tlacotepec de Benito Juárez, Puebla-Mexico, for the support provided to this research.

Conflicts of Interest

All authors declare no conflict of interest.

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Applsci 13 07428 g001 550
Figure 1. Diet mixtures: (A) Commercial feed®; (B) Sold-out substrate, wheat germ, and corn tortilla; (C) Sold-out substrate and wheat germ, and (D) Wheat germ and corn tortilla.
Figure 1. Diet mixtures: (A) Commercial feed®; (B) Sold-out substrate, wheat germ, and corn tortilla; (C) Sold-out substrate and wheat germ, and (D) Wheat germ and corn tortilla.
Applsci 13 07428 g001
Applsci 13 07428 g002 550
Figure 2. Analysis of color characteristics of the breast muscle (BM) and thigh muscle weight (TMW) at 24 h post-mortem of broilers fed diets containing sold-out substrate with different percentages of protein. Means with different letters indicate statistically significant differences by ANOVA and Tukey’s test (p < 0.05).
Figure 2. Analysis of color characteristics of the breast muscle (BM) and thigh muscle weight (TMW) at 24 h post-mortem of broilers fed diets containing sold-out substrate with different percentages of protein. Means with different letters indicate statistically significant differences by ANOVA and Tukey’s test (p < 0.05).
Applsci 13 07428 g002
Applsci 13 07428 g003 550
Figure 3. Proximal chemical analysis of breast muscle (MO) and thigh muscle weight (TMW) at 24 h post-mortem of broilers fed diets containing sold-out substrate with different percentages of protein. Means with different letters indicate statistically significant differences by ANOVA and Tukey’s test (p < 0.05).
Figure 3. Proximal chemical analysis of breast muscle (MO) and thigh muscle weight (TMW) at 24 h post-mortem of broilers fed diets containing sold-out substrate with different percentages of protein. Means with different letters indicate statistically significant differences by ANOVA and Tukey’s test (p < 0.05).
Applsci 13 07428 g003
Table
Table 1. Ingredients contained in the commercial feed® for the starter stage for broilers.
Table 1. Ingredients contained in the commercial feed® for the starter stage for broilers.
Ingredient(g)
Sorghum54.00
Soya flour21.00
Meat flour8.00
Peanut flour9.47
Soy oil5.23
Common iodized salt0.20
DL-Methionine0.25
Pecutrin1.42
Biolys0.43
Total (g)100.00
Crude protein (%)22.16
Table
Table 2. Experimental dietary ingredient content, metabolizable energy value, and crude protein percentage essential for broiler feeding.
Table 2. Experimental dietary ingredient content, metabolizable energy value, and crude protein percentage essential for broiler feeding.
IngredientTreatments
TCT1T2T3
Commercial feed® (g)100.00---
Wheat germ (g)-50.0050.0050.00
Sold-out substrate (g)-25.0050.00-
Corn tortilla (g)-25.00-50.00
Total (g)100.00100.00100.00100.00
Crude protein (%)20.1620.2619.6420.89
Carbohydrates (%)70.0062.8459.1966.50
Fat (%)8.506.195.536.85
Moisture (%)12.006.256.515.99
Crude fiber (%)8.008.7112.295.12
Ash (%)5.007.8212.313.34
Metabolizable energy (kcal/kg)3275.002946.743032.902861.10
Table
Table 3. Characteristics of sold-out substrate.
Table 3. Characteristics of sold-out substrate.
IndicatorsIn 100 g of Dry MatterUnits
Crude protein5.78(%)
Moisture 9.66
Ethereal extract0.37
Organic material63.00
Crude fiber16.04
Ash19.85
Ca6.24
Mg1.17
Na1.80
K2.75
P0.01
Total carbohydrates64.38
Fe765.00mg/kg
Cu3.00
Mn113.50
Zn29.50
Metabolizable energy2836.1Kcal/kg
Table
Table 4. Effect of sold-out substrate with different protein levels on production performance in broilers.
Table 4. Effect of sold-out substrate with different protein levels on production performance in broilers.
ParameterTCT1T2T3p-Value
Hematocrit (%)31.50 ± 1.50 b33.52 ± 0.5 a33.23 ± 0.00 a30.45 ± 0.00 b0.028
Cholesterol (mmol/L)3.43 ± 0.26 a2.81 ± 0.18 c3.13 ± 0.12 b3.40 ± 0.05 a0.014
Starting weight (kg)0.48 ± 0.01 a0.46 ± 0.02 b0.43 ± 0.01 c0.41 ± 0.00 d0.006
Final weight (kg)2.77 ± 0.30 a2.22 ± 0.25 b2.29 ± 0.18 b2.35 ± 0.40 b0.158
Overall weight gain (kg)2.33 ± 0.34 a1.76 ± 0.10 b1.85 ± 0.04 b1.93 ± 0.43 b0.019
Feed intake (kg)3.58 ± 0.13 a3.24 ± 0.14 b3.08 ± 0.10 b3.05 ± 0.17 b0.008
Water intake (L)10.07 ± 0.01 a9.34± 0.09 c9.70 ± 0.11 b7.47 ± 0.11 d0.003
FCR1.53 ± 0.10 a1.84 ± 1.10 a1.66 ± 0.15 a1.57 ± 0.94 a0.063
FCR: feed conversion ratio. Means with different letters indicate statistically significant differences by ANOVA and Tukey’s test (p < 0.05).
Table
Table 5. Effect of sold-out substrate with different levels of protein in carcass weight.
Table 5. Effect of sold-out substrate with different levels of protein in carcass weight.
TreatmentsParameter (g)
LWSWBWCWVCWwVFWVFWBMWTMWWWHNWGWLW
Control treatment
TC		2774265112325212214130475606262271804941
T122292117112202517169274274881771634345
T222942177116207917359814454.54971831624554
T32355225210321521797100344764911861644546
S.e.d.94.2793.315.1988.68203.805.7712.7063.5056.0312.7012.1242.885.77
p-value0.150.160.05 *0.190.140.09 *0.07 *0.390.140.02 *0.080.380.08 *
Abbreviations: Live weight (LW); slaughter weight (SW); bleeding weight (BW); carcass weight with viscera (CWV); carcass weight without viscera (CWwV); feather weight (FW); viscera fat weight (VFW); breast muscle weight (BMW); thigh muscle weight (TMW); wing weight (WW); head and neck weight (HNW); gizzard weight (GW), and liver weight (LW); S.e.d: standard error of deviation; * Indicates statistically significant differences by ANOVA and Tukey’s test (p < 0.05).
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Romero-Arenas, O.; Martínez Carrera, M.; Landeta-Cortés, G.; Rodríguez-Hernández, V.; Villa-Ruano, N.; Rivera, A. Pilot Study of Diet Supplemented with Sold-Out Substrate of Pleurotus ostreatus in the Feeding of Backyard Broilers. Appl. Sci. 2023, 13, 7428. https://doi.org/10.3390/app13137428

AMA Style

Romero-Arenas O, Martínez Carrera M, Landeta-Cortés G, Rodríguez-Hernández V, Villa-Ruano N, Rivera A. Pilot Study of Diet Supplemented with Sold-Out Substrate of Pleurotus ostreatus in the Feeding of Backyard Broilers. Applied Sciences. 2023; 13(13):7428. https://doi.org/10.3390/app13137428

Chicago/Turabian Style

Romero-Arenas, Omar, Maribel Martínez Carrera, Gerardo Landeta-Cortés, Victor Rodríguez-Hernández, Nemesio Villa-Ruano, and Antonio Rivera. 2023. "Pilot Study of Diet Supplemented with Sold-Out Substrate of Pleurotus ostreatus in the Feeding of Backyard Broilers" Applied Sciences 13, no. 13: 7428. https://doi.org/10.3390/app13137428

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