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Innovative dulce de leche made by sheep’s milk with and without the addition of sheep’s milk cream

Abstract

The objective of this work was to develop innovative formulations of dulce de leche made with sheep's milk with and without the addition of sheep’s milk cream and evaluate the shelf life of these products. Physicochemical and microbiological evaluations were performed on sheep’s pasteurized milk and cream, as well as on dulce de leche right after production and during the storage time. Addition of cream to sheep dulce de leche did not change the physicochemical and microbiological parameters between formulations. The dulce de leche showed dark color and adequate results of protein, fat, total and microbiological counts (thermotolerant coliforms, Staphylococcus aureus, Salmonella sp. in all evaluations, and total mesophilic aerobic up to 150 days of storage). Innovative formulations of dulce de leche made by sheep’s milk were developed, differentiated from those existing in the Brazilian market, but a specific legislation is required in Brazil focused on the specific characteristics and quality parameters of sheep’s milk and its derivatives.

Keywords:
sheep dairy products; shelf life; physicochemical and microbiological evaluation

1 Introduction

Sheep farming has grown in recent years in Brazil and the Brazilian sheep herd represents 22.11% of the sheep in the American continent in 2017 (Food and Agriculture Organization of the United Nations, 2019Food and Agriculture Organization of the United Nations – FAO. (2019). FAOSTAT. Rome. Retrieved from http://www.fao.org/faostat/en/#data/QA
http://www.fao.org/faostat/en/#data/QA...
). Climatic conditions in Southern Brazil favor the production and adaptation of several dairy sheep breeds and, consequently, their productivity, presenting a high economic potential for this sector (Munieweg et al., 2017Munieweg, F. R., Nespolo, C. R., Pinheiro, F. C., Gavião, E. R., Pinheiro, F. C., & Czarnobay, M. (2017). Qualidade do leite cru ovino armazenado sob refrigeração. Vigilância Sanitária em Debate, 5(1), 52-59. http://dx.doi.org/10.22239/2317-269x.00848.
http://dx.doi.org/10.22239/2317-269x.008...
). Sheep milk has higher macronutrient contents than bovine milk (Balthazar et al., 2017Balthazar, C. F., Pimentel, T. C., Ferrão, L. L., Almada, C. N., Santillo, A., Albenzio, M., Mollakhalili, N., Mortazavian, A. M., Nascimento, J. S., Silva, M. C., Freitas, M. Q., Sant’Ana, A. S., Granato, D., & Cruz, A. G. (2017). Sheep milk: physicochemical characteristics and relevance for functional food development. Comprehensive Reviews in Food Science and Food Safety, 16(2), 247-262. http://dx.doi.org/10.1111/1541-4337.12250.
http://dx.doi.org/10.1111/1541-4337.1225...
; Munieweg et al., 2017Munieweg, F. R., Nespolo, C. R., Pinheiro, F. C., Gavião, E. R., Pinheiro, F. C., & Czarnobay, M. (2017). Qualidade do leite cru ovino armazenado sob refrigeração. Vigilância Sanitária em Debate, 5(1), 52-59. http://dx.doi.org/10.22239/2317-269x.00848.
http://dx.doi.org/10.22239/2317-269x.008...
), but its composition might be influenced by animal’s race and health status, milking techniques, environment and processing technology, among other factors (Balthazar et al., 2017Balthazar, C. F., Pimentel, T. C., Ferrão, L. L., Almada, C. N., Santillo, A., Albenzio, M., Mollakhalili, N., Mortazavian, A. M., Nascimento, J. S., Silva, M. C., Freitas, M. Q., Sant’Ana, A. S., Granato, D., & Cruz, A. G. (2017). Sheep milk: physicochemical characteristics and relevance for functional food development. Comprehensive Reviews in Food Science and Food Safety, 16(2), 247-262. http://dx.doi.org/10.1111/1541-4337.12250.
http://dx.doi.org/10.1111/1541-4337.1225...
). The average composition of raw sheep milk produced in Southern Brazil ranges from 5.5 to 6.0% of protein, 5.9 to 7.3% of fat, 0.7 to 0.9% of ashes and 17.1 to 17.5% of total solids (Munieweg et al., 2017Munieweg, F. R., Nespolo, C. R., Pinheiro, F. C., Gavião, E. R., Pinheiro, F. C., & Czarnobay, M. (2017). Qualidade do leite cru ovino armazenado sob refrigeração. Vigilância Sanitária em Debate, 5(1), 52-59. http://dx.doi.org/10.22239/2317-269x.00848.
http://dx.doi.org/10.22239/2317-269x.008...
).

Sheep milk production in Brazil is concentrated in one company being responsible for all stages of production (Santos et al., 2016Santos, F. F., Nunes, R., & Gameiro, A. H. (2016). Notas sobre as transações no sistema agroindustrial do leite de ovelha no Brasil. Socioeconomia & Ciência Animal, 98, 1-6.). This raises the cost of milk production, reduces its commercialization and leads to the production of higher value added to derivatives such as cheese, yogurt and dulce de leche, allowing the maintenance of the farm and the expansion of dairy sheep farming (Santos et al., 2016Santos, F. F., Nunes, R., & Gameiro, A. H. (2016). Notas sobre as transações no sistema agroindustrial do leite de ovelha no Brasil. Socioeconomia & Ciência Animal, 98, 1-6.; Balthazar et al. 2017Balthazar, C. F., Pimentel, T. C., Ferrão, L. L., Almada, C. N., Santillo, A., Albenzio, M., Mollakhalili, N., Mortazavian, A. M., Nascimento, J. S., Silva, M. C., Freitas, M. Q., Sant’Ana, A. S., Granato, D., & Cruz, A. G. (2017). Sheep milk: physicochemical characteristics and relevance for functional food development. Comprehensive Reviews in Food Science and Food Safety, 16(2), 247-262. http://dx.doi.org/10.1111/1541-4337.12250.
http://dx.doi.org/10.1111/1541-4337.1225...
; Munieweg et al., 2017Munieweg, F. R., Nespolo, C. R., Pinheiro, F. C., Gavião, E. R., Pinheiro, F. C., & Czarnobay, M. (2017). Qualidade do leite cru ovino armazenado sob refrigeração. Vigilância Sanitária em Debate, 5(1), 52-59. http://dx.doi.org/10.22239/2317-269x.00848.
http://dx.doi.org/10.22239/2317-269x.008...
).

Dulce de leche is a traditional South American product prepared by slowly heating milk and sugar, with or without the addition of flavoring, until it reaches a concentration of approximately 68-70 °Brix (Brasil, 1997Brasil, Ministério de Estado da Agricultura, Pecuária e Abastecimento. (1997, September 8). Regulamento técnico para fixação de identidade e qualidade de doce de leite (Portaria nº 354, de 4 de setembro de 1997). Diário Oficial [da] República Federativa do Brasil.; Gaze et al., 2015Gaze, L. V., Costa, M. P., Monteiro, M. L. G., Lavorato, J. A. A., Conte, C. A. Jr., Raices, R. S. L., Cruz, A. G., & Freitas, M. Q. (2015). Dulce de Leche, a typical product of Latin America: characterisation by physicochemical, optical and instrumental methods. Food Chemistry, 169, 471-477. http://dx.doi.org/10.1016/j.foodchem.2014.08.017. PMid:25236253.
http://dx.doi.org/10.1016/j.foodchem.201...
; Francisquini et al., 2019Francisquini, J. A., Pereira, J. P. F., Pinto, M. S., Carvalho, A. F., Perrone, I. T., & Silva, P. H. F. (2019). Evolution of soluble solid content and evaporation rate curves during the manufacture of dulce de leche (dl). Food Science and Technology, 39(1), 78-82. http://dx.doi.org/10.1590/fst.26617.
http://dx.doi.org/10.1590/fst.26617...
). Brazilian law defines the characteristics and classification of dulce de leche (Brasil, 1996bBrasil, Ministério de Estado da Agricultura, Pecuária e Abastecimento. (1996b, December 17). Regulamento técnico MERCOSUL de identidade e qualidade de doce de leite (Resolução nº 137, de 13 de dezembro de 1996). Diário Oficial [da] República Federativa do Brasil., 1997Brasil, Ministério de Estado da Agricultura, Pecuária e Abastecimento. (1997, September 8). Regulamento técnico para fixação de identidade e qualidade de doce de leite (Portaria nº 354, de 4 de setembro de 1997). Diário Oficial [da] República Federativa do Brasil., 2001Brasil, Ministério da Saúde. Agência Nacional de Vigilância Sanitária. (2001, January 10). Aprova o regulamento técnico sobre padrões microbiológicos para alimentos (Resolução RDC nº 12, de 2 de janeiro de 2001). Diário Oficial [da] República Federativa do Brasil.), focused on products of bovine origin, and this dairy product from sheep milk could represent an economic alternative for producers and a differentiated option for consumers.

Derivatives made of sheep milk have been recently studied, including Ricotta cheese (Fusaro et al., 2019Fusaro, I., Giammarco, M., Chincarini, M., Vaintrub, M. O., Formigoni, A., Mammi, L. M. E., & Vignola, G. (2019). Fatty acids, health indices and sensory properties of Ricotta cheese from sheep fed three different diets. International Journal of Dairy Technology, 72(3), 427-434. http://dx.doi.org/10.1111/1471-0307.12613.
http://dx.doi.org/10.1111/1471-0307.1261...
), ice cream with probiotic (Balthazar et al., 2018bBalthazar, C. F., Silva, H. L. A., Esmerino, E. A., Rocha, R. S., Moraes, J., Carmo, M. A. V., Azevedo, L., Camps, I., K.D Abud, Y., Sant’Anna, C., Franco, R. M., Freitas, M. Q., Silva, M. C., Raices, R. S. L., Escher, G. B., Granato, D., Senaka Ranadheera, C., Nazarro, F., & Cruz, A. G. . (2018b). The addition of inulin and Lactobacillus casei 01 in sheep milk ice cream. Food Chemistry, 246, 464-472. http://dx.doi.org/10.1016/j.foodchem.2017.12.002. PMid:29291874.
http://dx.doi.org/10.1016/j.foodchem.201...
), beverage with strawberry (Balthazar et al., 2018aBalthazar, C. F., Santillo, A., Figliola, L., Silva, H. L. A., Esmerino, E. A., Freitas, M. Q., Cruz, A. G., & Albenzio, M. (2018a). Sensory evaluation of a novel prebiotic sheep milk strawberry beverage. Lebensmittel-Wissenschaft + Technologie, 98, 94-98. http://dx.doi.org/10.1016/j.lwt.2018.08.017.
http://dx.doi.org/10.1016/j.lwt.2018.08....
) and milk-juice beverage with fermented milk (Balthazar et al., 2019Balthazar, C. F., Santillo, A., Guimarães, J. T., Capozzi, V., Russo, P., Caroprese, M., Marino, R., Esmerino, E. A., Raices, R. S. L., Silva, M. C., Silva, H. L. A., Freitas, M. Q., Granato, D., Cruz, A. G., & Albenzio, M. (2019). Novel milk-juice beverage with fermented sheep milk and strawberry (Fragaria × ananassa): nutritional and functional characterization. Journal of Dairy Science, 102(12), 10724-10736. http://dx.doi.org/10.3168/jds.2019-16909. PMid:31521367.
http://dx.doi.org/10.3168/jds.2019-16909...
). Studies with sheep milk and derivatives have demonstrated that they are good sources of proteins and minerals (Balthazar et al., 2019Balthazar, C. F., Santillo, A., Guimarães, J. T., Capozzi, V., Russo, P., Caroprese, M., Marino, R., Esmerino, E. A., Raices, R. S. L., Silva, M. C., Silva, H. L. A., Freitas, M. Q., Granato, D., Cruz, A. G., & Albenzio, M. (2019). Novel milk-juice beverage with fermented sheep milk and strawberry (Fragaria × ananassa): nutritional and functional characterization. Journal of Dairy Science, 102(12), 10724-10736. http://dx.doi.org/10.3168/jds.2019-16909. PMid:31521367.
http://dx.doi.org/10.3168/jds.2019-16909...
), beneficial fatty acids such as α-linoleic acid and conjugated linoleic acid (Renes et al., 2018Renes, E., De la Fuente, F., Fernández, D., Tornadijo, M. E., & Fresno, J. M. (2018). Effect of feeding regimen on the fatty acid profile of sheep bulk tank milk. International Journal of Dairy Technology, 71(4), 857-867. http://dx.doi.org/10.1111/1471-0307.12553.
http://dx.doi.org/10.1111/1471-0307.1255...
; Fusaro et al., 2019Fusaro, I., Giammarco, M., Chincarini, M., Vaintrub, M. O., Formigoni, A., Mammi, L. M. E., & Vignola, G. (2019). Fatty acids, health indices and sensory properties of Ricotta cheese from sheep fed three different diets. International Journal of Dairy Technology, 72(3), 427-434. http://dx.doi.org/10.1111/1471-0307.12613.
http://dx.doi.org/10.1111/1471-0307.1261...
), in addition to their unique sensorial characteristics and functional properties (Balthazar et al., 2018aBalthazar, C. F., Santillo, A., Figliola, L., Silva, H. L. A., Esmerino, E. A., Freitas, M. Q., Cruz, A. G., & Albenzio, M. (2018a). Sensory evaluation of a novel prebiotic sheep milk strawberry beverage. Lebensmittel-Wissenschaft + Technologie, 98, 94-98. http://dx.doi.org/10.1016/j.lwt.2018.08.017.
http://dx.doi.org/10.1016/j.lwt.2018.08....
, bBalthazar, C. F., Silva, H. L. A., Esmerino, E. A., Rocha, R. S., Moraes, J., Carmo, M. A. V., Azevedo, L., Camps, I., K.D Abud, Y., Sant’Anna, C., Franco, R. M., Freitas, M. Q., Silva, M. C., Raices, R. S. L., Escher, G. B., Granato, D., Senaka Ranadheera, C., Nazarro, F., & Cruz, A. G. . (2018b). The addition of inulin and Lactobacillus casei 01 in sheep milk ice cream. Food Chemistry, 246, 464-472. http://dx.doi.org/10.1016/j.foodchem.2017.12.002. PMid:29291874.
http://dx.doi.org/10.1016/j.foodchem.201...
, 2019Balthazar, C. F., Santillo, A., Guimarães, J. T., Capozzi, V., Russo, P., Caroprese, M., Marino, R., Esmerino, E. A., Raices, R. S. L., Silva, M. C., Silva, H. L. A., Freitas, M. Q., Granato, D., Cruz, A. G., & Albenzio, M. (2019). Novel milk-juice beverage with fermented sheep milk and strawberry (Fragaria × ananassa): nutritional and functional characterization. Journal of Dairy Science, 102(12), 10724-10736. http://dx.doi.org/10.3168/jds.2019-16909. PMid:31521367.
http://dx.doi.org/10.3168/jds.2019-16909...
; Fusaro et al., 2019Fusaro, I., Giammarco, M., Chincarini, M., Vaintrub, M. O., Formigoni, A., Mammi, L. M. E., & Vignola, G. (2019). Fatty acids, health indices and sensory properties of Ricotta cheese from sheep fed three different diets. International Journal of Dairy Technology, 72(3), 427-434. http://dx.doi.org/10.1111/1471-0307.12613.
http://dx.doi.org/10.1111/1471-0307.1261...
).

The innovative character of sheep dairy products in Brazil occurs due to the use of raw sheep milk, which is unconventional and with a limited number of commercial products. The production of dulce de leche from this raw material and with the addition of sheep’s milk cream presents an innovative product to meet a strong demand for the gourmet and premium market. Thus, the objective of this work was to develop two innovative formulations of dulce de leche using sheep milk, with and without the addition of sheep milk cream, and to monitor the shelf life of these products through physical, chemical and microbiological evaluations.

2 Materials and methods

Sheep milk and cream were supplied by a dairy industry located in Southern Brazil that works with animals of the Lacaune breed. Sheep milk was previously skimmed to 3% of fat. Milk and cream were pasteurized and transported in plastic buckets for food use, placed in thermal boxes and transported under refrigeration to the laboratories. The ingredients such as crystallized sugar, glucose syrup and sodium bicarbonate were purchased on a local market.

Preliminary tests were performed to define the proportions of the raw materials and ingredients, as well as to the cooking times and inclusion of each component. The temperature during the preparation of the formulations was checked using a skewer thermometer CE F1007 (Incoterm®) and aliquots were collected to evaluate the total soluble solids (TSS) content in a model DR201-95 digital bench refractometer (A KrussOptronic®). Two formulations of dulce de leche were used: one without (DOC) and another with the addition (DWC) of sheep’s milk cream. The DWC formulation contained 3% of cream in partial substitution to the sheep milk. The other ingredients added were crystallized sugar, glucose syrup and sodium bicarbonate. The product was packaged while warm in sterilized glass jars (120 mL) with lids and stored at room temperature for 180 days, with random collection of three jars every 30 days for further evaluations.

Pasteurized milk and cream were evaluated upon arrival and the physicochemical analyzes included: protein, titratable acidity and pH analysis, as well as total dry extract, fat and cryoscopic index for pasteurized milk only. In the dulce de leche, water activity (Aw) and titratable acidity were evaluated, with periodic collections every 30 days, in addition to fat, protein and pH at 0, 90 and 180 days after production. Assessments of fat, protein, titratable acidity and cryoscopic index followed official methodologies (Brasil, 2017Brasil, Ministério da Agricultura, Pecuária e Abastecimento. (2017). Manual de métodos oficiais para análise de alimentos de origem animal (1. ed.). Brasília: MAPA.). The Aw was performed on an Aqualab 4TE meter (Decagon®) and the pH was determined on a pg1800 benchtop potentiometer (GGHAKA®) (Brasil, 2017Brasil, Ministério da Agricultura, Pecuária e Abastecimento. (2017). Manual de métodos oficiais para análise de alimentos de origem animal (1. ed.). Brasília: MAPA.). The values used as references were those established by the current Brazilian legislation (Brasil, 1996aBrasil, Ministério da Agricultura Pecuária e Abastecimento. (1996a, March 11). Regulamentos técnicos de identidade e qualidade dos produtos lácteos (Portaria nº 146, 7 de março de 1996). Diário Oficial [da] República Federativa do Brasil., bBrasil, Ministério de Estado da Agricultura, Pecuária e Abastecimento. (1996b, December 17). Regulamento técnico MERCOSUL de identidade e qualidade de doce de leite (Resolução nº 137, de 13 de dezembro de 1996). Diário Oficial [da] República Federativa do Brasil., 1997Brasil, Ministério de Estado da Agricultura, Pecuária e Abastecimento. (1997, September 8). Regulamento técnico para fixação de identidade e qualidade de doce de leite (Portaria nº 354, de 4 de setembro de 1997). Diário Oficial [da] República Federativa do Brasil., 2018Brasil, Ministério da Agricultura, Pecuária e Abastecimento. (2018, November 30). Regulamento técnico de identidade e qualidade do leite cru refrigerado (Instrução Normativa nº 76, 26 de novembro de 2018). Diário Oficial [da] República Federativa do Brasil.).

Microbiological analyzes on milk and cream were performed upon arrival and on dulce de leche every 30 days up to six months. Microbiological analyzes in all samples followed standardized methodologies and included: total mesophilic aerobic counts, total and thermotolerant coliforms (Brasil, 2017Brasil, Ministério da Agricultura, Pecuária e Abastecimento. (2017). Manual de métodos oficiais para análise de alimentos de origem animal (1. ed.). Brasília: MAPA.), S. aureus and Salmonella sp. (Silva et al., 2017Silva, N., Junqueira, V. C. A., Silveira, N. F. A., Taniwaki, M. H., Gomes, R. A. R., & Okazaki, M. M. (2017). Manual de métodos de análise microbiológica de alimentos e água (5. ed.). São Paulo: Varela.). Psychrotrophic microorganisms (Silva et al., 2017Silva, N., Junqueira, V. C. A., Silveira, N. F. A., Taniwaki, M. H., Gomes, R. A. R., & Okazaki, M. M. (2017). Manual de métodos de análise microbiológica de alimentos e água (5. ed.). São Paulo: Varela.) were determined only in milk and cream and the quantification of mold and yeast (Silva et al., 2017Silva, N., Junqueira, V. C. A., Silveira, N. F. A., Taniwaki, M. H., Gomes, R. A. R., & Okazaki, M. M. (2017). Manual de métodos de análise microbiológica de alimentos e água (5. ed.). São Paulo: Varela.) was performed in the dulce de leche. The values were compared with the limits established by the current Brazilian legislation (Brasil, 1996aBrasil, Ministério da Agricultura Pecuária e Abastecimento. (1996a, March 11). Regulamentos técnicos de identidade e qualidade dos produtos lácteos (Portaria nº 146, 7 de março de 1996). Diário Oficial [da] República Federativa do Brasil., bBrasil, Ministério de Estado da Agricultura, Pecuária e Abastecimento. (1996b, December 17). Regulamento técnico MERCOSUL de identidade e qualidade de doce de leite (Resolução nº 137, de 13 de dezembro de 1996). Diário Oficial [da] República Federativa do Brasil., 1997Brasil, Ministério de Estado da Agricultura, Pecuária e Abastecimento. (1997, September 8). Regulamento técnico para fixação de identidade e qualidade de doce de leite (Portaria nº 354, de 4 de setembro de 1997). Diário Oficial [da] República Federativa do Brasil., 2001Brasil, Ministério da Saúde. Agência Nacional de Vigilância Sanitária. (2001, January 10). Aprova o regulamento técnico sobre padrões microbiológicos para alimentos (Resolução RDC nº 12, de 2 de janeiro de 2001). Diário Oficial [da] República Federativa do Brasil., 2018Brasil, Ministério da Agricultura, Pecuária e Abastecimento. (2018, November 30). Regulamento técnico de identidade e qualidade do leite cru refrigerado (Instrução Normativa nº 76, 26 de novembro de 2018). Diário Oficial [da] República Federativa do Brasil.).

Color analyzes were performed with a portable measuring Chroma Meter CR-400 (Konica Minolta®) colorimeter, which works with three scales: L*, a* and b*. L* measures brightness and ranges from 0 for black to 100 for perfectly white surfaces. The a* scale measures the intensity of green (-) and red (+) color, while b* evaluates the transition from blue (-) to yellow (+) color (Ahmed et al., 2019Ahmed, E., Rita, R. P., Huq, S., Kabir, E., Rana, M. S., & Mazumder, M. A. R. (2019). Development of functional condensed milk from coconut milk and soy milk. Asian Food Science Journal, 6(4), 1-7. http://dx.doi.org/10.9734/AFSJ/2019/45269.
http://dx.doi.org/10.9734/AFSJ/2019/4526...
; Gaze et al., 2015Gaze, L. V., Costa, M. P., Monteiro, M. L. G., Lavorato, J. A. A., Conte, C. A. Jr., Raices, R. S. L., Cruz, A. G., & Freitas, M. Q. (2015). Dulce de Leche, a typical product of Latin America: characterisation by physicochemical, optical and instrumental methods. Food Chemistry, 169, 471-477. http://dx.doi.org/10.1016/j.foodchem.2014.08.017. PMid:25236253.
http://dx.doi.org/10.1016/j.foodchem.201...
). The colorimeter was previously calibrated and the analyzes were performed in triplicate. The total color difference (ΔE) between the two formulations was calculated as a way of assessing the differences noticeable to the human eye, considering the formula without cream (DOC) and with cream (DWC), using the formula that considers the variations of L*, a* and b*: ΔE* = [ΔL*2 + Δa*2 + Δb*2]1/2 (Gaze et al., 2015Gaze, L. V., Costa, M. P., Monteiro, M. L. G., Lavorato, J. A. A., Conte, C. A. Jr., Raices, R. S. L., Cruz, A. G., & Freitas, M. Q. (2015). Dulce de Leche, a typical product of Latin America: characterisation by physicochemical, optical and instrumental methods. Food Chemistry, 169, 471-477. http://dx.doi.org/10.1016/j.foodchem.2014.08.017. PMid:25236253.
http://dx.doi.org/10.1016/j.foodchem.201...
; Ahmed et al., 2019Ahmed, E., Rita, R. P., Huq, S., Kabir, E., Rana, M. S., & Mazumder, M. A. R. (2019). Development of functional condensed milk from coconut milk and soy milk. Asian Food Science Journal, 6(4), 1-7. http://dx.doi.org/10.9734/AFSJ/2019/45269.
http://dx.doi.org/10.9734/AFSJ/2019/4526...
).

Results were tabulated using a Microsoft Excel 2013 program, obtaining mean and standard deviation from the mean, and microbiological counts values converted to logarithms (log). Data were evaluated by the ASSISTAT 7.7 beta program, applying the analysis of variance followed by the Tukey test.

3 Results and discussion

The raw materials used in the production of dulce de leche were evaluated and pasteurized sheep milk showed 3.04 ± 0.03% of protein; 3.35 ± 0.05% of fat; 17.64 ± 2.35% of total dry extract; titratable acidity of 0.19 ± 0.01% in lactic acid; pH of 6.66 ± 0.05 and cryoscopic index of -0.518 ± 0.02 ºH. The fat content in pasteurized milk was lower than whole milk of Lacaune sheep, which has average contents above 5.8% (Munieweg et al., 2017Munieweg, F. R., Nespolo, C. R., Pinheiro, F. C., Gavião, E. R., Pinheiro, F. C., & Czarnobay, M. (2017). Qualidade do leite cru ovino armazenado sob refrigeração. Vigilância Sanitária em Debate, 5(1), 52-59. http://dx.doi.org/10.22239/2317-269x.00848.
http://dx.doi.org/10.22239/2317-269x.008...
; Tribst et al., 2020Tribst, A. A. L., Falcade, L. P., Leite, B. R. C. Jr., & Oliveira, M. M. (2020). Why are most physicochemical parameters not useful in predicting the quality of sheep milk? International Journal of Dairy Technology, 73(1), 292-295. http://dx.doi.org/10.1111/1471-0307.12657.
http://dx.doi.org/10.1111/1471-0307.1265...
), due to previous fat removal. Protein values of pasteurized sheep milk were lower than those observed in other studies with raw sheep milk, with averages of 4.8 to 6.0% (Balthazar et al., 2017Balthazar, C. F., Pimentel, T. C., Ferrão, L. L., Almada, C. N., Santillo, A., Albenzio, M., Mollakhalili, N., Mortazavian, A. M., Nascimento, J. S., Silva, M. C., Freitas, M. Q., Sant’Ana, A. S., Granato, D., & Cruz, A. G. (2017). Sheep milk: physicochemical characteristics and relevance for functional food development. Comprehensive Reviews in Food Science and Food Safety, 16(2), 247-262. http://dx.doi.org/10.1111/1541-4337.12250.
http://dx.doi.org/10.1111/1541-4337.1225...
; Munieweg et al., 2017Munieweg, F. R., Nespolo, C. R., Pinheiro, F. C., Gavião, E. R., Pinheiro, F. C., & Czarnobay, M. (2017). Qualidade do leite cru ovino armazenado sob refrigeração. Vigilância Sanitária em Debate, 5(1), 52-59. http://dx.doi.org/10.22239/2317-269x.00848.
http://dx.doi.org/10.22239/2317-269x.008...
; Renes et al., 2018Renes, E., De la Fuente, F., Fernández, D., Tornadijo, M. E., & Fresno, J. M. (2018). Effect of feeding regimen on the fatty acid profile of sheep bulk tank milk. International Journal of Dairy Technology, 71(4), 857-867. http://dx.doi.org/10.1111/1471-0307.12553.
http://dx.doi.org/10.1111/1471-0307.1255...
), probably due to differences in the processing with removal of components during skimming and heating in pasteurization. Both percentages of fat and protein in sheep milk evaluated were in accordance with the Brazilian legislation (Brasil, 2018Brasil, Ministério da Agricultura, Pecuária e Abastecimento. (2018, November 30). Regulamento técnico de identidade e qualidade do leite cru refrigerado (Instrução Normativa nº 76, 26 de novembro de 2018). Diário Oficial [da] República Federativa do Brasil.). The protein content of sheep's milk cream was lower than those found in bovine cream marketed in Brazil, with an average of 2.39% (Stephani et al., 2011Stephani, R., Neves, H. C., Neves, E. O., Souza, A. B., Perrone, I. T., & Silva, P. H. F. (2011). Caracterização físico-química do creme de leite UHT comercializado no Brasil. Revista do Instituto de Laticínios Cândido Tostes, 66(379), 25-29.) and the low efficiency of the skimming equipment may be the reason for this reduced percentage. Pasteurized sheep milk showed acidity above 0.14 to 0.18% of lactic acid defined by legislation (Brasil, 2018Brasil, Ministério da Agricultura, Pecuária e Abastecimento. (2018, November 30). Regulamento técnico de identidade e qualidade do leite cru refrigerado (Instrução Normativa nº 76, 26 de novembro de 2018). Diário Oficial [da] República Federativa do Brasil.), according to other authors (Balthazar et al., 2017Balthazar, C. F., Pimentel, T. C., Ferrão, L. L., Almada, C. N., Santillo, A., Albenzio, M., Mollakhalili, N., Mortazavian, A. M., Nascimento, J. S., Silva, M. C., Freitas, M. Q., Sant’Ana, A. S., Granato, D., & Cruz, A. G. (2017). Sheep milk: physicochemical characteristics and relevance for functional food development. Comprehensive Reviews in Food Science and Food Safety, 16(2), 247-262. http://dx.doi.org/10.1111/1541-4337.12250.
http://dx.doi.org/10.1111/1541-4337.1225...
; Munieweg et al., 2017Munieweg, F. R., Nespolo, C. R., Pinheiro, F. C., Gavião, E. R., Pinheiro, F. C., & Czarnobay, M. (2017). Qualidade do leite cru ovino armazenado sob refrigeração. Vigilância Sanitária em Debate, 5(1), 52-59. http://dx.doi.org/10.22239/2317-269x.00848.
http://dx.doi.org/10.22239/2317-269x.008...
; Tribst et al., 2020Tribst, A. A. L., Falcade, L. P., Leite, B. R. C. Jr., & Oliveira, M. M. (2020). Why are most physicochemical parameters not useful in predicting the quality of sheep milk? International Journal of Dairy Technology, 73(1), 292-295. http://dx.doi.org/10.1111/1471-0307.12657.
http://dx.doi.org/10.1111/1471-0307.1265...
). For sheep’s milk cream, the average acidity observed was below the maximum value allowed in legislation (0.20% of lactic acid) (Brasil, 1996aBrasil, Ministério da Agricultura Pecuária e Abastecimento. (1996a, March 11). Regulamentos técnicos de identidade e qualidade dos produtos lácteos (Portaria nº 146, 7 de março de 1996). Diário Oficial [da] República Federativa do Brasil.). The pH values of sheep raw milk and cream were similar to those found in bovine milk cream (Stephani et al., 2011Stephani, R., Neves, H. C., Neves, E. O., Souza, A. B., Perrone, I. T., & Silva, P. H. F. (2011). Caracterização físico-química do creme de leite UHT comercializado no Brasil. Revista do Instituto de Laticínios Cândido Tostes, 66(379), 25-29.) and lower than those observed in raw sheep milk (Munieweg et al., 2017Munieweg, F. R., Nespolo, C. R., Pinheiro, F. C., Gavião, E. R., Pinheiro, F. C., & Czarnobay, M. (2017). Qualidade do leite cru ovino armazenado sob refrigeração. Vigilância Sanitária em Debate, 5(1), 52-59. http://dx.doi.org/10.22239/2317-269x.00848.
http://dx.doi.org/10.22239/2317-269x.008...
; Tribst et al., 2020Tribst, A. A. L., Falcade, L. P., Leite, B. R. C. Jr., & Oliveira, M. M. (2020). Why are most physicochemical parameters not useful in predicting the quality of sheep milk? International Journal of Dairy Technology, 73(1), 292-295. http://dx.doi.org/10.1111/1471-0307.12657.
http://dx.doi.org/10.1111/1471-0307.1265...
) and sheep milk ice cream made with skimmed sheep milk added to sheep milk cream (Balthazar et al., 2018bBalthazar, C. F., Silva, H. L. A., Esmerino, E. A., Rocha, R. S., Moraes, J., Carmo, M. A. V., Azevedo, L., Camps, I., K.D Abud, Y., Sant’Anna, C., Franco, R. M., Freitas, M. Q., Silva, M. C., Raices, R. S. L., Escher, G. B., Granato, D., Senaka Ranadheera, C., Nazarro, F., & Cruz, A. G. . (2018b). The addition of inulin and Lactobacillus casei 01 in sheep milk ice cream. Food Chemistry, 246, 464-472. http://dx.doi.org/10.1016/j.foodchem.2017.12.002. PMid:29291874.
http://dx.doi.org/10.1016/j.foodchem.201...
). The cryoscopic index observed in pasteurized sheep milk was below the range recommended by law (-0.530 ° H to -0.555 °H) (Brasil, 2018Brasil, Ministério da Agricultura, Pecuária e Abastecimento. (2018, November 30). Regulamento técnico de identidade e qualidade do leite cru refrigerado (Instrução Normativa nº 76, 26 de novembro de 2018). Diário Oficial [da] República Federativa do Brasil.), the same reported in another study with raw sheep milk (Munieweg et al., 2017Munieweg, F. R., Nespolo, C. R., Pinheiro, F. C., Gavião, E. R., Pinheiro, F. C., & Czarnobay, M. (2017). Qualidade do leite cru ovino armazenado sob refrigeração. Vigilância Sanitária em Debate, 5(1), 52-59. http://dx.doi.org/10.22239/2317-269x.00848.
http://dx.doi.org/10.22239/2317-269x.008...
), and these differences may be due to the higher milk solids content of sheep as opposed to the legal limits set up for bovine milk (Brasil, 2018Brasil, Ministério da Agricultura, Pecuária e Abastecimento. (2018, November 30). Regulamento técnico de identidade e qualidade do leite cru refrigerado (Instrução Normativa nº 76, 26 de novembro de 2018). Diário Oficial [da] República Federativa do Brasil.).

Microbiological evaluation of pasteurized sheep milk resulted in the following counts: 9.92 ± 0.01 log CFU/mL for total mesophilic aerobic; 1.18 ± 0.01 log NMP/mL total coliforms; 0.96 ± 0.01 log NMP/mL of thermotolerant coliforms; 9.24 ± 0.17 log CFU/mL for psychrotrophs; absence of S. aureus at dilution 10-1 and Salmonella sp. in 25 mL of milk. The pasteurized sheep's milk cream contained 0.17 ± 0.06% of protein, titratable acidity of 0.08 ± 0.01% in lactic acid, and pH 6.75 ± 0.01. The mesophilic aerobic count in cream was 5.56 ± 0.01 log CFU/g; less than 0.48 log NMP/g for both total and thermotolerant coliforms; 5.83 ± 0.07 log CFU/g for psychrotrophs; absence of S. aureus at dilution 10-1 and Salmonella sp. in 25g. The mesophilic aerobic count in the cream and pasteurized milk exceeded the legal limits (Brasil, 1996aBrasil, Ministério da Agricultura Pecuária e Abastecimento. (1996a, March 11). Regulamentos técnicos de identidade e qualidade dos produtos lácteos (Portaria nº 146, 7 de março de 1996). Diário Oficial [da] República Federativa do Brasil.), as well as in other studies with raw milk of goats and sheep in Brazil (Agibert, 2013Agibert, S. A. C. (2013). Caracterização reológica, microbiológica, físico-química e sensorial de doce de leite caprino (Dissertação de mestrado). Universidade Federal do Rio de Janeiro, Rio de Janeiro.; Munieweg et al., 2017Munieweg, F. R., Nespolo, C. R., Pinheiro, F. C., Gavião, E. R., Pinheiro, F. C., & Czarnobay, M. (2017). Qualidade do leite cru ovino armazenado sob refrigeração. Vigilância Sanitária em Debate, 5(1), 52-59. http://dx.doi.org/10.22239/2317-269x.00848.
http://dx.doi.org/10.22239/2317-269x.008...
; Jiménez-Sobrino et al., 2018Jiménez-Sobrino, L., Garzón-Sigler, A., Pérez-Guzmán Palomares, M. D., García-Martínez, A., & Arias-Sánchez, R. (2018). Calidad microbiológica diferencial de la leche de oveja procedente de tanque. Revista Científica FCV-LUZ, 28(1), 11-18.), a fact not observed in European studies using sheep milk (Renes et al., 2018Renes, E., De la Fuente, F., Fernández, D., Tornadijo, M. E., & Fresno, J. M. (2018). Effect of feeding regimen on the fatty acid profile of sheep bulk tank milk. International Journal of Dairy Technology, 71(4), 857-867. http://dx.doi.org/10.1111/1471-0307.12553.
http://dx.doi.org/10.1111/1471-0307.1255...
). Brazilian legislation establishes maximum limits for pasteurized milk of 0.69 log NMP/mL for Enterobacteriaceae (Brasil, 2018Brasil, Ministério da Agricultura, Pecuária e Abastecimento. (2018, November 30). Regulamento técnico de identidade e qualidade do leite cru refrigerado (Instrução Normativa nº 76, 26 de novembro de 2018). Diário Oficial [da] República Federativa do Brasil.) and 0.60 log NMP/mL for thermotolerant coliforms (Brasil, 2001Brasil, Ministério da Saúde. Agência Nacional de Vigilância Sanitária. (2001, January 10). Aprova o regulamento técnico sobre padrões microbiológicos para alimentos (Resolução RDC nº 12, de 2 de janeiro de 2001). Diário Oficial [da] República Federativa do Brasil.), so pasteurized sheep's milk was above the allowed limit for this last evaluated agent. For pasteurized sour cream, the maximum limit for total coliforms is 1.0 log MPN/g (Brasil, 1996aBrasil, Ministério da Agricultura Pecuária e Abastecimento. (1996a, March 11). Regulamentos técnicos de identidade e qualidade dos produtos lácteos (Portaria nº 146, 7 de março de 1996). Diário Oficial [da] República Federativa do Brasil.) and for thermotolerant coliforms ranges from 0.48 log MPN/g (Brasil, 1996aBrasil, Ministério da Agricultura Pecuária e Abastecimento. (1996a, March 11). Regulamentos técnicos de identidade e qualidade dos produtos lácteos (Portaria nº 146, 7 de março de 1996). Diário Oficial [da] República Federativa do Brasil.) to 0.60 log MPN/g (Brasil, 2001Brasil, Ministério da Saúde. Agência Nacional de Vigilância Sanitária. (2001, January 10). Aprova o regulamento técnico sobre padrões microbiológicos para alimentos (Resolução RDC nº 12, de 2 de janeiro de 2001). Diário Oficial [da] República Federativa do Brasil.), therefore the raw material evaluated in the present study was in accordance with the established values. Several pathogenic and deteriorating microorganisms are mesophilic aerobics and coliform group is an indicator of hygienic-sanitary quality (Forsythe, 2013Forsythe, S. J. (2013). Microbiologia da segurança dos alimentos (2. ed.). Porto Alegre: Artmed.), and high counts may indicate hygiene failures during milking, transport and/or refrigerated milk storage (Munieweg et al., 2017Munieweg, F. R., Nespolo, C. R., Pinheiro, F. C., Gavião, E. R., Pinheiro, F. C., & Czarnobay, M. (2017). Qualidade do leite cru ovino armazenado sob refrigeração. Vigilância Sanitária em Debate, 5(1), 52-59. http://dx.doi.org/10.22239/2317-269x.00848.
http://dx.doi.org/10.22239/2317-269x.008...
; Jiménez-Sobrino et al., 2018Jiménez-Sobrino, L., Garzón-Sigler, A., Pérez-Guzmán Palomares, M. D., García-Martínez, A., & Arias-Sánchez, R. (2018). Calidad microbiológica diferencial de la leche de oveja procedente de tanque. Revista Científica FCV-LUZ, 28(1), 11-18.). The absence of S. aureus and Salmonella sp. in the evaluated raw materials is in accordance with the legal parameters (Brasil, 1996aBrasil, Ministério da Agricultura Pecuária e Abastecimento. (1996a, March 11). Regulamentos técnicos de identidade e qualidade dos produtos lácteos (Portaria nº 146, 7 de março de 1996). Diário Oficial [da] República Federativa do Brasil., 2001Brasil, Ministério da Saúde. Agência Nacional de Vigilância Sanitária. (2001, January 10). Aprova o regulamento técnico sobre padrões microbiológicos para alimentos (Resolução RDC nº 12, de 2 de janeiro de 2001). Diário Oficial [da] República Federativa do Brasil.) and this monitoring is important since they are pathogenic bacteria and are associated with contamination in dairy products (Forsythe, 2013Forsythe, S. J. (2013). Microbiologia da segurança dos alimentos (2. ed.). Porto Alegre: Artmed.). Psychrotrophic counts were high in pasteurized milk and sheep's milk cream, but there is no limit set by the Brazilian legislation. This group of microorganisms is common in products stored under refrigeration and has also been found in pasteurized bovine milk, raw sheep and goat milk (Agibert, 2013Agibert, S. A. C. (2013). Caracterização reológica, microbiológica, físico-química e sensorial de doce de leite caprino (Dissertação de mestrado). Universidade Federal do Rio de Janeiro, Rio de Janeiro.; Munieweg et al., 2017Munieweg, F. R., Nespolo, C. R., Pinheiro, F. C., Gavião, E. R., Pinheiro, F. C., & Czarnobay, M. (2017). Qualidade do leite cru ovino armazenado sob refrigeração. Vigilância Sanitária em Debate, 5(1), 52-59. http://dx.doi.org/10.22239/2317-269x.00848.
http://dx.doi.org/10.22239/2317-269x.008...
; Jiménez-Sobrino et al., 2018Jiménez-Sobrino, L., Garzón-Sigler, A., Pérez-Guzmán Palomares, M. D., García-Martínez, A., & Arias-Sánchez, R. (2018). Calidad microbiológica diferencial de la leche de oveja procedente de tanque. Revista Científica FCV-LUZ, 28(1), 11-18.). Sheep milk and sour cream were pasteurized to reduce microbial contamination (Forsythe, 2013Forsythe, S. J. (2013). Microbiologia da segurança dos alimentos (2. ed.). Porto Alegre: Artmed.), but this process was not enough in pasteurized sheep milk because high total and psychrotrophic mesophilic aerobic counts were observed. Brazilian sheep milk producers indicate the need for specific legislation for sheep milk, such as that defined for goat milk, due to the specificities of this raw material and the difficulties in adapting to the requirements established for bovine milk.

The results of protein, fat and pH contents of the dulce de leche are shown in Table 1.

Table 1
Fat, protein and pH contents of two formulations of dulce de leche with and without the addition of sheep's milk cream during storage.

Based on fat contents (Table 1) and the legislation (Brasil, 1997Brasil, Ministério de Estado da Agricultura, Pecuária e Abastecimento. (1997, September 8). Regulamento técnico para fixação de identidade e qualidade de doce de leite (Portaria nº 354, de 4 de setembro de 1997). Diário Oficial [da] República Federativa do Brasil.), dulce de leche without cream (DOC) was classified as a pure dulce de leche and the formulation with cream (DWC) was classified as dulce de leche with cream milk, compatible with the composition of each formulation. Other authors report average fat content in dulce de leche made of goat milk ranging from 16 to 20% (Agibert, 2013Agibert, S. A. C. (2013). Caracterização reológica, microbiológica, físico-química e sensorial de doce de leite caprino (Dissertação de mestrado). Universidade Federal do Rio de Janeiro, Rio de Janeiro.), 7.9% for sweetened condensed milk (Renhe et al., 2017Renhe, I. R. T., Pereira, D. B. C., Sá, J. F. O., Santos, M. C., Teodoro, V. A. M., Magalhães, F. A. R., Perrone, I. R., & Silva, P. H. F. (2017). Characterization of physicochemical composition, microbiology, sensory evaluation and microscopical attributes of sweetened condensed milk. Food Science and Technology, 38(2), 293-298. http://dx.doi.org/10.1590/1678-457x.34416.
http://dx.doi.org/10.1590/1678-457x.3441...
) and 3.6 to 7% in dulce de leche of bovine milk (Gaze et al., 2015Gaze, L. V., Costa, M. P., Monteiro, M. L. G., Lavorato, J. A. A., Conte, C. A. Jr., Raices, R. S. L., Cruz, A. G., & Freitas, M. Q. (2015). Dulce de Leche, a typical product of Latin America: characterisation by physicochemical, optical and instrumental methods. Food Chemistry, 169, 471-477. http://dx.doi.org/10.1016/j.foodchem.2014.08.017. PMid:25236253.
http://dx.doi.org/10.1016/j.foodchem.201...
; Silva et al., 2015Silva, F. L., Ferreira, H. A. L., Souza, A. B., Almeida, D. F., Stephani, R., Pirozi, M. R., Carvalho, A. F., & Perrone, I. T. (2015). Production of dulce de leche: the effect of starch addition. Lebensmittel-Wissenschaft + Technologie, 62(1), 417. http://dx.doi.org/10.1016/j.lwt.2014.10.062.
http://dx.doi.org/10.1016/j.lwt.2014.10....
). The structure and distribution of fat globules can be affected by some processes such as heat treatment, agitation and centrifugation employed to obtain milk cream (Balthazar et al., 2017Balthazar, C. F., Pimentel, T. C., Ferrão, L. L., Almada, C. N., Santillo, A., Albenzio, M., Mollakhalili, N., Mortazavian, A. M., Nascimento, J. S., Silva, M. C., Freitas, M. Q., Sant’Ana, A. S., Granato, D., & Cruz, A. G. (2017). Sheep milk: physicochemical characteristics and relevance for functional food development. Comprehensive Reviews in Food Science and Food Safety, 16(2), 247-262. http://dx.doi.org/10.1111/1541-4337.12250.
http://dx.doi.org/10.1111/1541-4337.1225...
). The DOC showed higher amounts (p < 0.05) of protein compared to DWC at 180 days of storage (Table 1) and formulations with sheep's milk cream contained protein levels above the minimum required by the legislation (Brasil, 1997Brasil, Ministério de Estado da Agricultura, Pecuária e Abastecimento. (1997, September 8). Regulamento técnico para fixação de identidade e qualidade de doce de leite (Portaria nº 354, de 4 de setembro de 1997). Diário Oficial [da] República Federativa do Brasil.). In the DWC formulation, there was a partial replacement of the pasteurized sheep's milk by pasteurized sheep's milk cream, which resulted on a lower protein content due to lower amount of protein in the cream compared to the milk (0.17% and 3.04%, respectively). Protein percentages in the DOC and DWC formulations were similar to those found in other studies with dulce de leche from goat milk (Agibert, 2013Agibert, S. A. C. (2013). Caracterização reológica, microbiológica, físico-química e sensorial de doce de leite caprino (Dissertação de mestrado). Universidade Federal do Rio de Janeiro, Rio de Janeiro.), from goat milk with chia flour (Chaves et al., 2018Chaves, M. A., Souza, A. H. P., Colla, E., Bittenccourt, P. R. S., & Matsushita, M. (2018). Influences of chia flour and the concentration of total solids on the characteristics of ‘dulce de leche’ from goat milk. Food Science and Technology, 38(Suppl. 1), 338-344. http://dx.doi.org/10.1590/1678-457x.22017.
http://dx.doi.org/10.1590/1678-457x.2201...
) and from bovine milk (Silva et al., 2015Silva, F. L., Ferreira, H. A. L., Souza, A. B., Almeida, D. F., Stephani, R., Pirozi, M. R., Carvalho, A. F., & Perrone, I. T. (2015). Production of dulce de leche: the effect of starch addition. Lebensmittel-Wissenschaft + Technologie, 62(1), 417. http://dx.doi.org/10.1016/j.lwt.2014.10.062.
http://dx.doi.org/10.1016/j.lwt.2014.10....
). Mean pH values ranged from 6.34 to 6.53, with no significant differences between formulations or sampling times (Table 1). These values were higher than those found in bovine dulce de leche, with the addition of goat's whey and with different concentrations of umbu pulp, whose pH ranged from 4.40 to 5.49 (Silva et al., 2011Silva, M. S. S., Figueirêdo, R. M. F., Queiroz, A. J. M., & Santiago, V. M. S. (2011). Avaliação físico-química e sensorial de doces cremosos produzidos com soro de leite de cabra, leite de vaca e polpa de umbu. Revista Brasileira de Produtos Agroindustriais, 13(4), 397-410. http://dx.doi.org/10.15871/1517-8595/rbpa.v13n4p397-410.
http://dx.doi.org/10.15871/1517-8595/rbp...
), similar to those observed in commercial bovine dulce de leche (6.14 to 6.37) (Gaze et al., 2015Gaze, L. V., Costa, M. P., Monteiro, M. L. G., Lavorato, J. A. A., Conte, C. A. Jr., Raices, R. S. L., Cruz, A. G., & Freitas, M. Q. (2015). Dulce de Leche, a typical product of Latin America: characterisation by physicochemical, optical and instrumental methods. Food Chemistry, 169, 471-477. http://dx.doi.org/10.1016/j.foodchem.2014.08.017. PMid:25236253.
http://dx.doi.org/10.1016/j.foodchem.201...
) and lower than those of buffalo dulce de leche (7.02 and 6.87) (Jacob et al., 2018Jacob, V. R., Roque, C. M., Silva, A. S. L., Neve, K. A. L., & Otani, F. S. (2018). Aspectos de qualidade físico-química de doce de leite de búfalas da raça Murrah, a partir de leite fresco e armazenado. Agroecossistemas, 9(2), 288-298. http://dx.doi.org/10.18542/ragros.v9i2.5122.
http://dx.doi.org/10.18542/ragros.v9i2.5...
). Considering the observed pH values, the formulations of the present work can be classified as low acid foods. which increases the risk of contamination by pathogenic microorganisms (Forsythe, 2013Forsythe, S. J. (2013). Microbiologia da segurança dos alimentos (2. ed.). Porto Alegre: Artmed.).

The evaluation of water activity and titratable acidity in dulce de leche made by sheep's milk are presented in Figure 1. The Aw results of dulce de leche ranged from 0.71 to 0.82 (Figure 1A), with significantly (p < 0.05) higher values at times 0 and 90 days for both formulations. Studies with bovine dulce de leche showed Aw values between 0.85 and 0.91 for industrialized formulations and condensed sweetened milk (Ferreira et al., 2012Ferreira, L. O., Pereira, P. A. P., Maria, J., & Pinto, S. M. (2012). Avaliação das características de qualidade de doces de leite comerciais. Revista do Instituto de Latícinios Cândido Tostes, 67(387), 5-11. http://dx.doi.org/10.5935/2238-6416.20120044.
http://dx.doi.org/10.5935/2238-6416.2012...
; Renhe et al., 2017Renhe, I. R. T., Pereira, D. B. C., Sá, J. F. O., Santos, M. C., Teodoro, V. A. M., Magalhães, F. A. R., Perrone, I. R., & Silva, P. H. F. (2017). Characterization of physicochemical composition, microbiology, sensory evaluation and microscopical attributes of sweetened condensed milk. Food Science and Technology, 38(2), 293-298. http://dx.doi.org/10.1590/1678-457x.34416.
http://dx.doi.org/10.1590/1678-457x.3441...
) and these variations may occur with dulce de leche, due to heating that can concentrate the ingredients and increase gradually the amount of insoluble solids (Francisquini et al., 2019Francisquini, J. A., Pereira, J. P. F., Pinto, M. S., Carvalho, A. F., Perrone, I. T., & Silva, P. H. F. (2019). Evolution of soluble solid content and evaporation rate curves during the manufacture of dulce de leche (dl). Food Science and Technology, 39(1), 78-82. http://dx.doi.org/10.1590/fst.26617.
http://dx.doi.org/10.1590/fst.26617...
). The titratable acidity in the products ranged from 0.41 to 0.71% of lactic acid in the DOC formulation and from 0.47 to 0.69% lactic acid in the DWC (Figure 1B). There was a gradual increase in acidity in both formulations during storage, with significant differences (p < 0.05) on day 180 of storage compared to the initial time, but without variation between DOC and DWC at any of the sampling times. Studies report 3.02% acidity in lactic acid of fresh milk of buffalo and 3.6% in dulce de leche stored for nine months (Jacob et al., 2018Jacob, V. R., Roque, C. M., Silva, A. S. L., Neve, K. A. L., & Otani, F. S. (2018). Aspectos de qualidade físico-química de doce de leite de búfalas da raça Murrah, a partir de leite fresco e armazenado. Agroecossistemas, 9(2), 288-298. http://dx.doi.org/10.18542/ragros.v9i2.5122.
http://dx.doi.org/10.18542/ragros.v9i2.5...
), much higher values than those found in fresh dulce de leche of this study. The acidity values in seven brands of dulce de leche marketed in Brazil showed values of 0.23 to 0.50% acidity in lactic acid (Gaze et al., 2015Gaze, L. V., Costa, M. P., Monteiro, M. L. G., Lavorato, J. A. A., Conte, C. A. Jr., Raices, R. S. L., Cruz, A. G., & Freitas, M. Q. (2015). Dulce de Leche, a typical product of Latin America: characterisation by physicochemical, optical and instrumental methods. Food Chemistry, 169, 471-477. http://dx.doi.org/10.1016/j.foodchem.2014.08.017. PMid:25236253.
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), which are much lower than those observed in the present study.

Figure 1
Evaluation of (A) water activity and (B) titratable acidity in two formulations of dulce de leche with and without the addition of sheep's milk cream during storage. The gray line represents the formulation DOC (dairy without cream) and the black line indicates the DWC (dulce de leche with cream) (n = 3).

The results of water activity of dulce de leche in the present study showed values below the minimum limit for multiplication of pathogenic bacteria, mainly due to the addition of sugar that acts to reduce the amount of free water in the food, however, the titratable acidity was low for microbial control to occur (Forsythe, 2013Forsythe, S. J. (2013). Microbiologia da segurança dos alimentos (2. ed.). Porto Alegre: Artmed.).

The quantification of the total mesophilic aerobic group over the storage period can be seen in Figure 2 and mold and yeast counts are shown in Table 2. The DOC formulation had counts between 1.52 to 9.42 log CFU/g, while DWC ranged from 1.52 to 7.49 log CFU/g, with significant differences (p < 0.05) at 180 days for both formulations compared to the other sampling times (Figure 2). The increase in the counting of this group after 150 days of storage might be associated with a process of deterioration, since these products were stored at room temperature, with favorable conditions for the growth of mesophilic microorganisms (Forsythe, 2013Forsythe, S. J. (2013). Microbiologia da segurança dos alimentos (2. ed.). Porto Alegre: Artmed.). High counts of these microorganisms were also found in bovine dulce de leche in Lavras, MG (Southeast of Brazil), with an average count of 5.86 log CFU/g (Oliveira et al., 2012Oliveira, A. R. C., Oliveira, R. M. E., Abreu, P. S., Ferreira, L. O., Domiciniano, D., & Pinto, S. M. (2012). Qualidade microbiológica de doces de leite comercializados no sul de Minas Gerais. Revista do Instituto de Latícinios Cândido Tostes, 67(388), 11-14. http://dx.doi.org/10.5935/2238-6416.20120059.
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) as well as for sweetened condensed milk samples, ranging from 1.0 and 6.2 log CFU/g (Renhe et al., 2017Renhe, I. R. T., Pereira, D. B. C., Sá, J. F. O., Santos, M. C., Teodoro, V. A. M., Magalhães, F. A. R., Perrone, I. R., & Silva, P. H. F. (2017). Characterization of physicochemical composition, microbiology, sensory evaluation and microscopical attributes of sweetened condensed milk. Food Science and Technology, 38(2), 293-298. http://dx.doi.org/10.1590/1678-457x.34416.
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). Although Brazilian legislation does not set limits for this microbial class in dulce de leche, monitoring of total mesophilic aerobes provides information on product contamination and may assist in defining the shelf life of a new product (Forsythe, 2013Forsythe, S. J. (2013). Microbiologia da segurança dos alimentos (2. ed.). Porto Alegre: Artmed.).

Figure 2
Total mesophilic aerobic count in both formulations of sheep's milk with and without the addition of sheep’s milk cream during storage. The gray line represents the formulation DOC (dairy without cream) and black indicates the formulation DWC (dairy with cream) (n = 3).
Table 2
Mold and yeast count of two formulations of dulce de leche with and without the addition of sheep’s milk during storage.

Beside this, both formulations of dulce de leche had counts of less than 0.48 log NMP/g for total and thermotolerant coliforms and absence of Salmonella sp. and S. aureus, which indicates that the samples are appropriate to the legislation considering these microbiological parameters (Brasil, 1996bBrasil, Ministério de Estado da Agricultura, Pecuária e Abastecimento. (1996b, December 17). Regulamento técnico MERCOSUL de identidade e qualidade de doce de leite (Resolução nº 137, de 13 de dezembro de 1996). Diário Oficial [da] República Federativa do Brasil., 1997Brasil, Ministério de Estado da Agricultura, Pecuária e Abastecimento. (1997, September 8). Regulamento técnico para fixação de identidade e qualidade de doce de leite (Portaria nº 354, de 4 de setembro de 1997). Diário Oficial [da] República Federativa do Brasil., 2001Brasil, Ministério da Saúde. Agência Nacional de Vigilância Sanitária. (2001, January 10). Aprova o regulamento técnico sobre padrões microbiológicos para alimentos (Resolução RDC nº 12, de 2 de janeiro de 2001). Diário Oficial [da] República Federativa do Brasil.). The low values quantified for the coliform group indicate that the processing and handling conditions were adequate, and the non-detection of Salmonella sp. and S. aureus emphasizes the safety of the food produced (Forsythe, 2013Forsythe, S. J. (2013). Microbiologia da segurança dos alimentos (2. ed.). Porto Alegre: Artmed.). Milk pasteurization and the cooking process while making the dulce de leche contribute to the destruction of microorganisms (Forsythe, 2013Forsythe, S. J. (2013). Microbiologia da segurança dos alimentos (2. ed.). Porto Alegre: Artmed.), and these steps were effective for the control of these microorganisms.

Mold and yeast counts presented values of 3.86 and 4.29 log CFU/g for the DOC and DWC formulations, respectively, at 180 days of storage (Table 2). In only two DWC sampling, the maximum limit for mold and yeast was not exceeded, while in DOC the counts were above the limit allowed by law (Brasil, 1996bBrasil, Ministério de Estado da Agricultura, Pecuária e Abastecimento. (1996b, December 17). Regulamento técnico MERCOSUL de identidade e qualidade de doce de leite (Resolução nº 137, de 13 de dezembro de 1996). Diário Oficial [da] República Federativa do Brasil., 1997Brasil, Ministério de Estado da Agricultura, Pecuária e Abastecimento. (1997, September 8). Regulamento técnico para fixação de identidade e qualidade de doce de leite (Portaria nº 354, de 4 de setembro de 1997). Diário Oficial [da] República Federativa do Brasil.) during the whole storage period. Previous work on goat dulce de leche, with or without whey (Agibert, 2013Agibert, S. A. C. (2013). Caracterização reológica, microbiológica, físico-química e sensorial de doce de leite caprino (Dissertação de mestrado). Universidade Federal do Rio de Janeiro, Rio de Janeiro.), have found mold and yeast counts lower than 2.0 log CFU/g allowed. On the other hand, counts above 3.0 log CFU/g were reported in bovine dulce de leche marketed in Lavras, MG (Southeast of Brazil) (Oliveira et al., 2012Oliveira, A. R. C., Oliveira, R. M. E., Abreu, P. S., Ferreira, L. O., Domiciniano, D., & Pinto, S. M. (2012). Qualidade microbiológica de doces de leite comercializados no sul de Minas Gerais. Revista do Instituto de Latícinios Cândido Tostes, 67(388), 11-14. http://dx.doi.org/10.5935/2238-6416.20120059.
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). In some of these formulations, the low counts for mold and yeast may have been influenced by the addition of preservatives allowed in the preparation of dulce de leche (Brasil, 2010Brasil, Ministério de Estado da Agricultura, Pecuária e Abastecimento. (2010, September 10). Altera o subitem 5.2 do Anexo da Portaria MAPA nº 354, de 4 de setembro de 1997 (Portaria nº 855, de 27 de setembro de 2010). Diário Oficial [da] República Federativa do Brasil.), considered the factor responsible for controlling the presence of fungi in dulce de leche formulated with or without sucrose (Milagres et al., 2010Milagres, M. P., Dias, G., Magalhães, M. A., Silva, M. O., & Ramos, A. M. (2010). Análise físico-química e sensorial de doce de leite produzido sem adição de sacarose. Revista Ceres, 57(4), 439-445. http://dx.doi.org/10.1590/S0034-737X2010000400001.
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). However, preservatives were not used in the formulations of the present study. Water activity results in sheep dulce de leche (Figure 1A) showed values from 0.71 to 0.82, which are limiting for pathogenic and some spoilage microorganisms, but the increase in sugar concentration would only inactivate microbial multiplication in Aw below 0.70 (Forsythe, 2013Forsythe, S. J. (2013). Microbiologia da segurança dos alimentos (2. ed.). Porto Alegre: Artmed.). The quantification of molds and yeasts, above the values allowed by legislation may indicate flaws in the processing and demonstrated how easy it is for fungi to grow in foods with low water activity, with potential risk to consumer health, since many fungi species produce mycotoxins (Oliveira et al., 2012Oliveira, A. R. C., Oliveira, R. M. E., Abreu, P. S., Ferreira, L. O., Domiciniano, D., & Pinto, S. M. (2012). Qualidade microbiológica de doces de leite comercializados no sul de Minas Gerais. Revista do Instituto de Latícinios Cândido Tostes, 67(388), 11-14. http://dx.doi.org/10.5935/2238-6416.20120059.
http://dx.doi.org/10.5935/2238-6416.2012...
; Forsythe, 2013Forsythe, S. J. (2013). Microbiologia da segurança dos alimentos (2. ed.). Porto Alegre: Artmed.). The preparation of DOC and DWC involved pasteurization and cooling of raw materials, concentration by addition of sugar and heat, and the use of good processing practices. All these processes help in the prevention and conservation of food, with destruction of pathogenic bacteria and reduction of microbial load (Oliveira et al., 2012Oliveira, A. R. C., Oliveira, R. M. E., Abreu, P. S., Ferreira, L. O., Domiciniano, D., & Pinto, S. M. (2012). Qualidade microbiológica de doces de leite comercializados no sul de Minas Gerais. Revista do Instituto de Latícinios Cândido Tostes, 67(388), 11-14. http://dx.doi.org/10.5935/2238-6416.20120059.
http://dx.doi.org/10.5935/2238-6416.2012...
; Forsythe, 2013Forsythe, S. J. (2013). Microbiologia da segurança dos alimentos (2. ed.). Porto Alegre: Artmed.). under storage at room temperature. However, the presence of molds and yeasts above the levels allowed by the legislation reveals failures that occurred in any of the production or sealing steps of the packaging, allowing the passage of oxygen and contaminants, although the non-use of chemical preservatives in sheep's milk dulce de leche.

Color results of the formulations can be seen in Table 3, without significant differences (p < 0.05) between formulations or storage periods.

Table 3
Color analysis on the L*, a* and b* scales for both formulations of dulce de leche with and without the addition of sheep’s milk during storage.

The L* values in the DOC and DWC were below 50 (Table 3), the midpoint of this scale, indicating a tendency of dark staining of the samples. In studies with dulce de leche, the brightness values were between 48 and 68 (Ferreira et al., 2012Ferreira, L. O., Pereira, P. A. P., Maria, J., & Pinto, S. M. (2012). Avaliação das características de qualidade de doces de leite comerciais. Revista do Instituto de Latícinios Cândido Tostes, 67(387), 5-11. http://dx.doi.org/10.5935/2238-6416.20120044.
http://dx.doi.org/10.5935/2238-6416.2012...
; Agibert, 2013Agibert, S. A. C. (2013). Caracterização reológica, microbiológica, físico-química e sensorial de doce de leite caprino (Dissertação de mestrado). Universidade Federal do Rio de Janeiro, Rio de Janeiro.; Silva et al., 2015Silva, F. L., Ferreira, H. A. L., Souza, A. B., Almeida, D. F., Stephani, R., Pirozi, M. R., Carvalho, A. F., & Perrone, I. T. (2015). Production of dulce de leche: the effect of starch addition. Lebensmittel-Wissenschaft + Technologie, 62(1), 417. http://dx.doi.org/10.1016/j.lwt.2014.10.062.
http://dx.doi.org/10.1016/j.lwt.2014.10....
; Gaze et al., 2015Gaze, L. V., Costa, M. P., Monteiro, M. L. G., Lavorato, J. A. A., Conte, C. A. Jr., Raices, R. S. L., Cruz, A. G., & Freitas, M. Q. (2015). Dulce de Leche, a typical product of Latin America: characterisation by physicochemical, optical and instrumental methods. Food Chemistry, 169, 471-477. http://dx.doi.org/10.1016/j.foodchem.2014.08.017. PMid:25236253.
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), higher than those found in the present work. In dulce de leche containing umbu pulp, the measurements ranged from 39.0 to 47.9 (Silva et al., 2011Silva, M. S. S., Figueirêdo, R. M. F., Queiroz, A. J. M., & Santiago, V. M. S. (2011). Avaliação físico-química e sensorial de doces cremosos produzidos com soro de leite de cabra, leite de vaca e polpa de umbu. Revista Brasileira de Produtos Agroindustriais, 13(4), 397-410. http://dx.doi.org/10.15871/1517-8595/rbpa.v13n4p397-410.
http://dx.doi.org/10.15871/1517-8595/rbp...
) and the values ranged between 41.8 and 43.3 for dulce de leche from goat milk and with chia flour (Chaves et al., 2018Chaves, M. A., Souza, A. H. P., Colla, E., Bittenccourt, P. R. S., & Matsushita, M. (2018). Influences of chia flour and the concentration of total solids on the characteristics of ‘dulce de leche’ from goat milk. Food Science and Technology, 38(Suppl. 1), 338-344. http://dx.doi.org/10.1590/1678-457x.22017.
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), close to the dulce de leche of the present study. The results of the a* scale were all positives, with no significant differences between formulations (p < 0.05), demonstrating a tendency to red (Table 3). The trend for this color was also observed in goat dulce de leche (Agibert, 2013Agibert, S. A. C. (2013). Caracterização reológica, microbiológica, físico-química e sensorial de doce de leite caprino (Dissertação de mestrado). Universidade Federal do Rio de Janeiro, Rio de Janeiro.; Chaves et al., 2018Chaves, M. A., Souza, A. H. P., Colla, E., Bittenccourt, P. R. S., & Matsushita, M. (2018). Influences of chia flour and the concentration of total solids on the characteristics of ‘dulce de leche’ from goat milk. Food Science and Technology, 38(Suppl. 1), 338-344. http://dx.doi.org/10.1590/1678-457x.22017.
http://dx.doi.org/10.1590/1678-457x.2201...
), commercial bovine dulce de leche (Gaze et al., 2015Gaze, L. V., Costa, M. P., Monteiro, M. L. G., Lavorato, J. A. A., Conte, C. A. Jr., Raices, R. S. L., Cruz, A. G., & Freitas, M. Q. (2015). Dulce de Leche, a typical product of Latin America: characterisation by physicochemical, optical and instrumental methods. Food Chemistry, 169, 471-477. http://dx.doi.org/10.1016/j.foodchem.2014.08.017. PMid:25236253.
http://dx.doi.org/10.1016/j.foodchem.201...
) and bovine dulce de leche with different concentrations of umbu pulp (Silva et al., 2011Silva, M. S. S., Figueirêdo, R. M. F., Queiroz, A. J. M., & Santiago, V. M. S. (2011). Avaliação físico-química e sensorial de doces cremosos produzidos com soro de leite de cabra, leite de vaca e polpa de umbu. Revista Brasileira de Produtos Agroindustriais, 13(4), 397-410. http://dx.doi.org/10.15871/1517-8595/rbpa.v13n4p397-410.
http://dx.doi.org/10.15871/1517-8595/rbp...
). The b* values indicated that both formulations of dulce de leche showed higher wavelength reflection associated with the yellow color (Table 3). These positive results were also identified by other authors (Agibert, 2013Agibert, S. A. C. (2013). Caracterização reológica, microbiológica, físico-química e sensorial de doce de leite caprino (Dissertação de mestrado). Universidade Federal do Rio de Janeiro, Rio de Janeiro.; Chaves et al., 2018Chaves, M. A., Souza, A. H. P., Colla, E., Bittenccourt, P. R. S., & Matsushita, M. (2018). Influences of chia flour and the concentration of total solids on the characteristics of ‘dulce de leche’ from goat milk. Food Science and Technology, 38(Suppl. 1), 338-344. http://dx.doi.org/10.1590/1678-457x.22017.
http://dx.doi.org/10.1590/1678-457x.2201...
; Gaze et al., 2015Gaze, L. V., Costa, M. P., Monteiro, M. L. G., Lavorato, J. A. A., Conte, C. A. Jr., Raices, R. S. L., Cruz, A. G., & Freitas, M. Q. (2015). Dulce de Leche, a typical product of Latin America: characterisation by physicochemical, optical and instrumental methods. Food Chemistry, 169, 471-477. http://dx.doi.org/10.1016/j.foodchem.2014.08.017. PMid:25236253.
http://dx.doi.org/10.1016/j.foodchem.201...
; Ferreira et al., 2012Ferreira, L. O., Pereira, P. A. P., Maria, J., & Pinto, S. M. (2012). Avaliação das características de qualidade de doces de leite comerciais. Revista do Instituto de Latícinios Cândido Tostes, 67(387), 5-11. http://dx.doi.org/10.5935/2238-6416.20120044.
http://dx.doi.org/10.5935/2238-6416.2012...
; Silva et al., 2011Silva, M. S. S., Figueirêdo, R. M. F., Queiroz, A. J. M., & Santiago, V. M. S. (2011). Avaliação físico-química e sensorial de doces cremosos produzidos com soro de leite de cabra, leite de vaca e polpa de umbu. Revista Brasileira de Produtos Agroindustriais, 13(4), 397-410. http://dx.doi.org/10.15871/1517-8595/rbpa.v13n4p397-410.
http://dx.doi.org/10.15871/1517-8595/rbp...
, 2015Silva, F. L., Ferreira, H. A. L., Souza, A. B., Almeida, D. F., Stephani, R., Pirozi, M. R., Carvalho, A. F., & Perrone, I. T. (2015). Production of dulce de leche: the effect of starch addition. Lebensmittel-Wissenschaft + Technologie, 62(1), 417. http://dx.doi.org/10.1016/j.lwt.2014.10.062.
http://dx.doi.org/10.1016/j.lwt.2014.10....
). The total color difference (ΔE) between the formulations indicated values of 0.96 at initial time, 2.13 at 90 days, and 1.88 at 180 days of storage. According to Gaze et al. (2015)Gaze, L. V., Costa, M. P., Monteiro, M. L. G., Lavorato, J. A. A., Conte, C. A. Jr., Raices, R. S. L., Cruz, A. G., & Freitas, M. Q. (2015). Dulce de Leche, a typical product of Latin America: characterisation by physicochemical, optical and instrumental methods. Food Chemistry, 169, 471-477. http://dx.doi.org/10.1016/j.foodchem.2014.08.017. PMid:25236253.
http://dx.doi.org/10.1016/j.foodchem.201...
, total color differences up to 1.5 are considered small, as observed at baseline, and ΔE of 1.5 to 3.0 between formulations indicate that they are distinct, which occurred at 90 and 180 days of storage. The brightness difference (ΔL*) showed negative results in all periods evaluated, indicating that the DWC formulation was slightly brighter than the DOC, but without significant variations (p < 0.05). The fact that there were no significant variations between both formulations of dulce de leche elaborated in the present study regarding their color, demonstrates that the technology used in the production of both was similar and that the cream added to the dulce de leche (DWC) did not interfere in this parameter. The initial acidity of milk, the amount and timing of the addition of sodium bicarbonate, and the initial and final soluble solids content may be some of the triggering factors for differences in the coloration of dulce de leche (Ferreira et al., 2012Ferreira, L. O., Pereira, P. A. P., Maria, J., & Pinto, S. M. (2012). Avaliação das características de qualidade de doces de leite comerciais. Revista do Instituto de Latícinios Cândido Tostes, 67(387), 5-11. http://dx.doi.org/10.5935/2238-6416.20120044.
http://dx.doi.org/10.5935/2238-6416.2012...
), a fact not observed in the present study.

In the next step of the study, we will conduct the sensory analysis of the dulce de leche formulations to verify the acceptance of these differentiated products in relation to those existing on the market. Acceptance sensory tests can evaluate parameters by a numerical scale (Balthazar et al., 2018aBalthazar, C. F., Santillo, A., Figliola, L., Silva, H. L. A., Esmerino, E. A., Freitas, M. Q., Cruz, A. G., & Albenzio, M. (2018a). Sensory evaluation of a novel prebiotic sheep milk strawberry beverage. Lebensmittel-Wissenschaft + Technologie, 98, 94-98. http://dx.doi.org/10.1016/j.lwt.2018.08.017.
http://dx.doi.org/10.1016/j.lwt.2018.08....
; Fusaro et al., 2019Fusaro, I., Giammarco, M., Chincarini, M., Vaintrub, M. O., Formigoni, A., Mammi, L. M. E., & Vignola, G. (2019). Fatty acids, health indices and sensory properties of Ricotta cheese from sheep fed three different diets. International Journal of Dairy Technology, 72(3), 427-434. http://dx.doi.org/10.1111/1471-0307.12613.
http://dx.doi.org/10.1111/1471-0307.1261...
) combined with relevance coefficients for each sensory parameter (Heydari et al., 2018Heydari, S., Amiri‐Rigi, A., Ehsani, M. R., Mohammadifar, M. A., Khorshidian, N., Koushki, M. R., & Mortazavian, A. M. (2018). Rheological behaviour, sensory properties and syneresis of probiotic yoghurt supplemented with various prebiotics. International Journal of Dairy Technology, 71(S1), 175-184. http://dx.doi.org/10.1111/1471-0307.12491.
http://dx.doi.org/10.1111/1471-0307.1249...
) and descriptive analysis can be used to identify the relationship between products and their sensory attributes, which must be performed by consumers with high sensory acuity in dairy assessment (Balthazar et al., 2018aBalthazar, C. F., Santillo, A., Figliola, L., Silva, H. L. A., Esmerino, E. A., Freitas, M. Q., Cruz, A. G., & Albenzio, M. (2018a). Sensory evaluation of a novel prebiotic sheep milk strawberry beverage. Lebensmittel-Wissenschaft + Technologie, 98, 94-98. http://dx.doi.org/10.1016/j.lwt.2018.08.017.
http://dx.doi.org/10.1016/j.lwt.2018.08....
; Chetachukwu et al., 2019Chetachukwu, A. S., Thongraung, C., & Yupanqui, C. T. (2019). Development of reduced‐fat coconut yoghurt: physicochemical, rheological, microstructural and sensory properties. International Journal of Dairy Technology, 72(4), 524-535. http://dx.doi.org/10.1111/1471-0307.12600.
http://dx.doi.org/10.1111/1471-0307.1260...
; Fusaro et al., 2019Fusaro, I., Giammarco, M., Chincarini, M., Vaintrub, M. O., Formigoni, A., Mammi, L. M. E., & Vignola, G. (2019). Fatty acids, health indices and sensory properties of Ricotta cheese from sheep fed three different diets. International Journal of Dairy Technology, 72(3), 427-434. http://dx.doi.org/10.1111/1471-0307.12613.
http://dx.doi.org/10.1111/1471-0307.1261...
). The use of the projective techniques has been applied to evaluate the sensorial perception of Brazilian consumers about healthy, differentiated or unconventional products in the market. These techniques, such as free word association and Haire’s shopping list methods, can be useful to comprehend the perceptions and beliefs about food products and can be applied in the development or reformulation of products and marketing strategies (Judacewski et al., 2019Judacewski, P., Los, P. R., Lima, L. S., Alberti, A., Zielinski, A. A. F., & Nogueira, A. (2019). Perceptions of Brazilian consumers regarding white mould surface‐ripened cheese using free word association. International Journal of Dairy Technology, 72(4), 585-590. http://dx.doi.org/10.1111/1471-0307.12649.
http://dx.doi.org/10.1111/1471-0307.1264...
; Pinto et al., 2018Pinto, L. P. F., Silva, H. L. A., Kuriya, S. P., Maçaira, P. M., Oliveira, F. L. C., Cruz, A. G., Esmerino, E. A., & Freitas, M. Q. (2018). Understanding perceptions and beliefs about different types of fermented milks through the application of projective techniques: a case study using haire’s shopping list and free word association. Journal of Sensory Studies, 33(3), e12326. http://dx.doi.org/10.1111/joss.12326.
http://dx.doi.org/10.1111/joss.12326...
). For similar purpose, consumer-based methodologies are gaining popularity in the sensorial science to improve the novel dairy development and to expand efficient marketing strategies of commercialization of these products. The preferred attribute elicitation methodology demonstrated to be useful to assess the perceptions of consumers of dairy product samples, and it did not need training, resulting in suitable reliability with a low number of consumers (Soares et al., 2019Soares, E. K. B., Silva, R., Silva, W. P., Kuriya, S. P., Maçaira, P. M., Oliveira, F. L. C., Silva, M. A. A. P., Pimentel, T. C., Freitas, M. Q., Cruz, A. G., & Esmerino, E. A. (2019). An intra-cultural investigation in Brazil using Coalho cheese and preferred attribute elicitation. Journal of Sensory Studies, 53(1), e12543. http://dx.doi.org/10.1111/joss.12543.
http://dx.doi.org/10.1111/joss.12543...
). The check-all-that-apply questionnaire allows assessors to select all appropriate sensory characteristics for describing a product and contributes to optimizing the different dairy foods with similar or complex sensory characteristics (Torres et al., 2017Torres, F. R., Esmerino, E. A., Thomas Carr, B., Ferrão, L. L., Granato, D., Pimentel, T. C., Bolini, H. M. A., Freitas, M. Q., & Cruz, A. G. (2017). Rapid consumer-based sensory characterization of requeijão cremoso, a spreadable processed cheese: Performance of new statistical approaches to evaluate check-all-that-apply data. Journal of Dairy Science, 100(8), 6100-6110. http://dx.doi.org/10.3168/jds.2016-12516.
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). All of these methods can contribute so that the dulce de leche developed in the present study have consumption potential in the Brazilian market.

4 Conclusions

Addition of cream to sheep dulce de leche did not change the physicochemical and microbiological parameters between formulations, that showed dark color, and sheep dulce de leche were stable up to 150 days of storage for most of the evaluated parameters, except for mold and yeast over the evaluated period. Innovative formulations of dulce de leche made by sheep's milk were developed, differentiated from those existing in the Brazilian market, but a specific legislation for sheep's milk and its derivatives is necessary in Brazil to establish the identity and quality of these products.

Acknowledgements

We thank the National Council for Scientific and Technological Development (CNPq), Brazil, Research and Innovation Support Foundation of Santa Catarina State (FAPESC), Santa Catarina, Brazil, and Academic Development Program (PDA) of Federal University of Pampa (UNIPAMPA), Rio Grande do Sul, Brazil.

  • Practical Application: The development of sheep’s dulce de leche formulations with and without the addition of sheep’s milk cream contributes to the diversification and innovation of production in dairy, especially from milk of small ruminants. The definition of conditions of processing and verification of quality during storage demonstrate diversification potential for small-scale production.

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Publication Dates

  • Publication in this collection
    22 June 2020
  • Date of issue
    June 2021

History

  • Received
    10 Mar 2020
  • Accepted
    15 Apr 2020
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