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Cuban Journal of Agricultural Science

Print version ISSN 0864-0408On-line version ISSN 2079-3480

Cuban J. Agric. Sci. vol.49 no.4 Mayabeque Oct.-Dec. 2015




Evaluation of a probiotic mixture in the started birds feeding of heavy pure breeds B4 in a production unit


Evaluación de una mezcla probiótica en la alimentación de aves de inicio de líneas puras pesadas B4 en una unidad de producción



Marlen Rodríguez,I Grethel Milián,I Ana J. Rondón,I R. Bocourt,II A., Beruvidez,I Elayne Crespo,III

ICentro de Estudios Biotecnológicos, Universidad de Matanzas, Matanzas, C.P. 44740, Cuba.
IIInstituto de Ciencia Animal, San José de las Lajas, Mayabeque, C.P. 3200, Cuba.
IIIEmpresa Genética Avícola y Pie de Cría, Matanzas, C.P. 10400, Cuba.




In order to evaluate the effect of the inclusion of a probiotic mixture (bio-preparation) based on Lactobacillus salivariusC65 and Bacillus subtilis E44 strains on the diet of started birds of heavy pure breeds, a research was conducted at the Unidad Genética Avícola “Granma” from Pedro Betancourt municipality, Matanzas. The study was carried out in a period of 35 days, between December2013 and January 2014.The design was completely randomized and two treatments were evaluated: T1) basal diet (control) and T2) basal diet plus bio-preparations mixture, in ratio 1:1. A total of 600 heavy pure breeds B4 birds were used, (300 per treatment) with an average weight of 42 grams at birth. They stayed on floor under similar management and feeding conditions. The liveweight, intake, conversion in weight per kilogram of intake food, mortality and birds viability was determined. The animals that received the probiotic mixture had a high liveweight during the rearing, except in the first two weeks, in which there were not differences between treatments. There were differences (P<0. 05) in the conversion per bird and the mortality percentage and viability, in favor of the treatment with the probiotic mixture. It concludes that it is possible to use this bio -preparation in the started birds feeding of heavy pure breeds, since the best biometric results were found in the group of birds treated with the animal additive.

Key words: probiotic mixture, heavy pure breeds.


Para evaluar el efecto de inclusión de una mezcla probiótica (biopreparado) basada en las cepas Lactobacillus salivarius C65 y Bacillus subtilis E44, en la dieta de aves de inicio de líneas puras pesadas,  se realizó una investigación en la Unidad Genética Avícola “Granma”, del municipio Pedro Betancourt, Matanzas, en un período de 35 d, entre diciembre de 2013 y enero de 2014. El diseño fue completamente aleatorizado y se evaluaron dos tratamientos: T1) dieta basal (control) y T2) dieta basal más la mezcla de biopreparados, en proporción 1:1. Se utilizaron 600 aves de la línea Pura Pesada B4, (300 por tratamiento) con peso promedio de 42 g al nacer. Se  alojaron en piso, en similares condiciones de manejo y alimentación. Se determinó el peso vivo, el consumo, la conversión en peso por kilogramo de alimento consumido, la mortalidad y la viabilidad de las aves. Los animales que recibieron la mezcla probiótica tuvieron  mayor peso vivo en toda la crianza, excepto en las primeras dos semanas, en la que no se manifestaron diferencias entre tratamientos. Hubo diferencias (P < 0.05) en la conversión por ave, el porcentaje de mortalidad y viabilidad, a favor del tratamiento con la mezcla probiótica. Se concluye que es posible utilizar este biopreparado en la alimentación de aves de inicio de líneas puras pesadas, ya que los mejores resultados se encontraron en el grupo de aves tratadas con el aditivo zootécnico.

Palabras clave: mezcla probiótica, líneas puras pesadas.




The introduction of new products and technologies for the obtaining of healthy foods, which allows high productions with an appropriate sustainability, is a priority in the politics for animal production in Cuba. In this context, many authors agree in that the probiotics could be useful in the improvement of zootechnical indicators in animals of economic interest (Pérez et al. 2012 and Milián et al. 2014).

The microorganisms more used as probiotics are the lactic acid bacteria (especially Lactobacillus and Bifidobacterium), yeasts (mainly the Saccharomyces genus) and bacteria from Bacillus genus and their endospores. All of them are part of bio-preparations that are in the international market (González-Núñez et al. 2013 and Carro et al. 2014).

The probiotic mixtures are constituted by two or more of these microorganisms, generally isolated from the digestive tract of healthy adult animals (Brizuela, 2011).Today it is recognized the importance and possible effectiveness of biotic therapy( probiotics and prebiotics) as medical tool in animal production(Garcia et al. 2012).

In Cuba, the Empresa Genética y Pie de Cría has as main social object the maintenance and improvement of the heavy pure breeds. With its production guarantees the replacement of all basic birds from the national system and prevents the country has to invest to import replacement regularly. This allows guaranteeing poultry genetic basis and the existing breeds, besides to favor the raising of meat and eggs yield, as well as the production and marketing of the heavy foundation stock in Poultry Enterprises (Godínez 2012). 

 The objective of this research was to evaluate the effect of the inclusion of a probiotic mixture (bio-preparation) based on Lactobacillus salivarius C65 and Bacillus subtilis E44 strains on the diet of started birds of heavy pure breeds, B4.



Preparation of the bacterial bio-preparation. From the Lactobacillus salivariusC65 and Bacillus subtilis E44 strains, 20L of probiotic mixture were elaborated. For these flasks with 1L of Mann, Rogosa, Sharper (MRS) media and nutrient medium for Lactobacillus and Bacillus were prepared, respectively. They were inoculated and cultured at 37 ºC during 18h.Then, was added 250 mL of inoculums in four Erlenmeyer of 6L, which contained 5L of medium for Lactobacillus salivarius or MCLs growth (Rondón 2009). For Bacillus growth the Bacillus subtilis (MCBs) growth medium described by Milian (2009) was used. Both cultures were stayed at 37 ºC during 20h, under static conditions for Lactobacillus and with agitation at 250 rpm for Bacillus. After this time, viable counts and pH measure were carried out, to check the bio-preparation quality. They were packaged in sterile flasks of 1L, with rubber top and were conserved at 5 ºC until their use. The cultures were mixed into equal parts at the time of the application.

Experimental conditions. Treatments. The experiment was conducted at the Unidad Genética Avícola “Granma” from Pedro Betancourt municipality, Matanzas. The evaluation of the in vivo bio-preparation was carried out during 35 days between December/2013 and January/2014. The design was completely randomized with two treatments: T1) basal diet (control) and T2) basal diet plus bio-preparations mixture, in ratio 1:1.A total of 600 heavy pure breeds B4 birds were used, distributed in 300 birds per

Birds management. The animals management was homogeneous in both groups, coinciding with the stated in the Technical Instructive No 7 UCAN -IIA (1998) for the evaluated species and category. The bio-preparation was given in the ration of the T2 group, in the morning twice a week. It was manually mixed with the diet, at a concentration of 109 UFC.mL-1, which is equivalent to 109 endospores of Bacillus g-1 concentrate. The dose was fitted according to the criteria proposed by Mutus et al. (2006) and Milián (2009).

Evaluation of the bio-preparation effect on started birds from heavy pure breeds B4, under production conditions. To evaluate the in vivo probiotic effect of the microbial biopreparation, the average liveweight of birds per treatment was determined weekly. The food intake and the conversion in weight per kilogram of food intake were determined. The mortality percentage was also calculated during the weeks of the experiment. It was also taking into account all parameters evaluated as a result of the biometric performed at 35 days. These indicators were calculated as described in the Technical Instructive for broiler rearing (UCAN- IIA, 1998).

Statistical processing of the in vivo evaluation. To analyze the results, the statistical Software INFOSTAT version 1 was used (Balzarini et al. 2001). For the statistical treatment of data variance analysis with completely randomized design were carried out and to verify significant differences between treatments the Duncan (1995) comparison test was used for a 95% reliability. The mortality and viability values were transformed to



The liveweight variation per week of rearing with the use of probiotic mixture is showed in table 1.

The birds that received the additive in diet exceed those of the control treatment in this indicator. There were significant differences (P<0.01) in favor of this treatment (T2), from the third evaluated week.

When probiotics are incorporated in the food as additive, an eubiosis state occurs and promotes a physiological effect on the organism, more than its nutritive value (Khasefidi and Goorchi 2006).The probiotics positive effects not only occur at the gastrointestinal tract (GIT), but are reflected in animal results as liveweight gain and the feed conversion improvement (Gil and Gil- Turnes 2005).

Khaksefidi and Ghoorchi (2006) obtained weight increase in chickens after supplying in diets, Bacillus subtilis cultures at 50 mg.Kg-1 of concentrate. These authors refers that differences (P<0.01) in the animals daily liveweight were presented.

When performing the corresponding biometric at 35 days of age, significant differences between treatments were observed (table 2).

The use of L. salivarius and B. subtilis mixture in birds diets represented an improvement in all the evaluated indicators. The liveweight at the end of the rearing and meat conversion per each kilogram of food intake, was better regarding the animals that did not received this microbial mixture.

The indicators evaluated in both groups are above the technical instructive indicators, used as comparison criteria. This supposed a positive control under very controlled conditions for the facilities of the Unión de Empresas del Combinado Avícola Nacional (UECAN).

The viability of birds was one of the parameters taken into account in health indicators during the experimental stage. In this case, the higher viability percentage corresponds to the birds treated with the bio-preparation. The group of birds that did not intake the probiotic mixture, not only resulted with lower viability, but they were not able to reach the percentage required by the standard line. According to Kizerwetter-Swida and Binek (2009), probiotics microorganisms, when acting, maintain a beneficial microflora in the animal and exclude those potentially pathogenic microorganisms. This supposes to be the cause for which the group of treated birds reaches better results.

Lan et al. (2003) informed that the supply of probiotic bacteria (L. salivarius and L. agilis) produce an increase of broilers liveweight. Other authors as Mohan et al. (1996), Zulkifli et al. (2000) and Balevi et al. (2001) also observed improvements in weight increase of the birds that were treated with probiotics.

Milián (2009) and Rondón (2009) evaluated the effect of different strains of Bacillus subtilis and Lactobacillus salivarius respectively, on productive indicators of broilers. These authors observed a better productive response in the treated animals regarding those of the control group. These results refers that probiotics, when stabilizing the microbiological system in the digestive tract, improve the nutrients digestibility and provide a range of higher absorption, when introducing an anabolism that promotes body weight gain with decreasing feed conversion.

Years later, a Bacillus subtilis E44 (subtilprobio) strains was evaluated in birds of heavy pure breeds E1 (González 2012) and, a Lactobacillus salivarius(Probiolactil) strain in birds of heavy pure breeds P8 (Suarez, 2013). As a result it was observed that the birds which ingested probiotic bacteria had better productive results than the control animals. This result agrees with those reached in this research, but in birds of heavy pure breeds B4, treated with the mixture of probiotic strains previously mentioned.

These results could be related to the benefits provided by probiotic microorganisms when favoring digestion, absorption and nutrients use. The use of probiotics bio-preparation in broilers is a routine practice in modern poultry production which provides favorable results (Lima 2007) and coincides with those of this research.

A naturalistic and update approach of probiotics conceives them as natural products which are used as growth promoters in animals, so that its use allows to obtain higher yields, higher immune resistant, reduced or non amount of pathogens in the GIT and lower antibiotic residues and other substances in the final products.

The bio-preparation that was used in the experiment could influence in the improvement of liveweight, conversion, uniformity and the higher number of selected birds, according to the standard criteria.



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Received: November 25, 2015
Accepted: January 6, 2016



Marlen Rodríguez, Centro de Estudios Biotecnológicos, Universidad de Matanzas, Matanzas, C.P. 44740, Cuba. Email:

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