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  • Effects of corn gluten feed inclusion at graded levels in a corn-soybean diet on the ileal and fecal digestibility of growing pigs

    • 1Email author,
    • 2 and
    • 1
    Journal of Animal Science and Biotechnology20145:40

    https://doi.org/10.1186/2049-1891-5-40

    • Received: 27 March 2014
    • Accepted: 11 August 2014
    • Published:

    Abstract

    Background

    This study aimed to determine the effect of the inclusion of corn gluten feed (CGF) on the apparent and standardized ileal digestibility of protein and amino acids and the apparent ileal and total tract digestibility of energy in growing pigs. The study was performed using 16 barrows (weight, 45.3 ± 4.5 kg) that were fitted with a T cannula at the terminal ileum. There were four treatments: a corn-soybean diet without CGF and three corn-soybean diets containing increasing levels of CGF (65, 130, and 195 g/kg). Data were analyzed according to a randomized complete block design, four blocks with four pigs each (one pig per treatment). The trend of the response (linear or quadratic) was determined using orthogonal contrasts, and when a linear effect was determined, a linear equation was obtained.

    Results

    The results showed that the inclusion up to 195 g/kg of CGF in the corn-soybean diet did not diminish the ileal digestibility (apparent and standardized) of protein and amino acids (P > 0.05), except that of phenylalanine, cystine, and proline. A linear decrease (P < 0.05) per gram of CGF added to the diet in the apparent and standardized ileal digestibility of phenylalanine (0.011 and 0.015 percentage units, respectively), cystine (0.048 and 0.043 percentage units, respectively), and proline (0.045 and 0.047 percentage units, respectively) was noted. Similarly, ileal digestibility of dry matter and energy were adversely affected (reduced by 0.028 and 0.025 percentage units, respectively, per gram of CGF increment in the diet). A significant (P < 0.05) linear reduction in total tract digestibility with increase in CGF amount in the diet was observed for energy (0.027 percentage units), dry matter (0.027 percentage units), crude protein (0.020 percentage units), and neutral detergent fiber (0.041 percentage units) per gram of CGF added to the diet.

    Conclusion

    CGF did not affect the ileal digestibility of protein and most amino acids but reduced the ileal and total tract digestibility of energy.

    Keywords

    • Amino acids
    • Corn gluten feed
    • Energy
    • Ileal digestibility
    • Pigs
    • Total tract digestibility

    Background

    Corn (Zea mays) is the most harvested cereal worldwide [1]. Most of the corn produced is processed to extract flour, syrup, sweeteners, starches, oils, and ethanol. When wet milling is used to process corn, the germ is separated from the kernel, processed to obtain edible oil, and the resulting flour is mixed with the bran to produce corn gluten feed (CGF) [2], which has traditionally been considered a protein feed [3], although it is rich in fiber [4, 5]. Therefore, the inclusion of CGF in pig diets has been limited, since fiber can affect directly [6, 7] and indirectly [8, 9] the digestibility of amino acids by increasing endogenous protein losses. Thus, this study aimed to determine the effects of inclusion of CGF in a corn-soybean meal diet on the apparent and standardized ileal digestibility of amino acids and apparent total tract digestibility of energy in growing pigs. We hypothesized that the inclusion of CGF decreases the ileal digestibility of amino acids and energy and the total tract digestibility of energy.

    Materials and methods

    This study was approved by the Scientific Associate Technical Group Committee of CENID Physiology. The animals used in this study were cared for in accordance with the guidelines issued by the Mexican Official Standard for the Production, Protection and Use of Lab Animals [10] and the guidelines of the International Guiding Principles for Biomedical Research Involving Animals [11]. The study was performed at the experimental farm of CENID-Physiology.

    Animals

    Sixteen barrows (Duroc × Landrace) weighing 45.3 ± 4.5 kg were used. The animals were placed in individual metabolic cages provided with a self-feeder and a low-pressure drinking nipple in a temperature controlled room at 20 ± 2°C. Animals were fed twice daily at 0800 h and 1800 h and had free access to water. A T cannula was fitted at the terminal ileum of each animal as previously described [12]. After surgery, therapeutic treatment (penicillin, 600,000 IU; streptomycin, 750 mg; oxytetracycline, 500 mg) was administered for 3 d; the post-surgery period lasted 21 d. From the second day after the post-surgery period, pigs began to receive 100 g of feed, which was increased by 100 g/day until the feed intake reached the level before surgery. During the experimental period the pigs were fed at 2.5-times the maintenance requirement of digestible energy of 460 kJ/kg BW0.75[13]. Pigs had free access to water.

    Experimental diets

    The experimental diets were produced using corn, soybean meal, and CGF (Table 1). Four diets were formulated (Table 2) with increasing levels of CGF (0, 65, 130, and 195 g/kg of diet) at the expenses of corn and soybean meal. Corn oil was included at a rate of 40 g/kg. Salt, vitamins, and minerals were included at the levels that met or exceeded the National Research Council (NRC) nutritional requirements [5]; chromic oxide was added at a rate of 3 g/kg of feed as an inert marker [14]. The total fecal collection was performed daily for 5 d to achieve the required chromic oxide rate. Ferric oxide was added (3 g/kg of diet) to the first meal of the fifth day (to mark the start of the collection period) as well as to the first meal of the eleventh day (to mark the end of the collection period) [15].
    Table 1

    Chemical composition of raw materials as fed-basis (g/kg)

    Item

    SBM1

    YC

    CGF

    Dry matter

    899.5

    900.3

    891.2

    Crude protein

    468.1

    108.5

    208.4

    NDF2

    89.5

    119.7

    338.7

    ADF3

    57.4

    45.0

    106.5

    Amino acids

       

    Alanine

    24.8

    7.2

    17.1

    Aspartic acid

    67.5

    9.7

    17.3

    Arginine

    42.6

    8.2

    12.1

    Cystine

    8.3

    2.7

    4.3

    Glutamic acid

    111.7

    20.8

    45.3

    Glycine

    24.1

    6.1

    12.2

    Histidine

    10.1

    4.7

    7.1

    Isoleucine

    24.5

    3.3

    7.6

    Leucine

    44.2

    10.1

    23.8

    Lysine

    30.3

    2.8

    5.4

    Methionine

    7.8

    2.0

    3.4

    Phenylalanine

    28.9

    4.7

    9.7

    Proline

    31.1

    8.8

    16.6

    Serine

    30.7

    5.7

    11.5

    Threonine

    27.9

    5.6

    12.1

    Tyrosine

    20.8

    3.9

    8.1

    Valine

    25.5

    5.6

    12.1

    1Raw materials: SBM = soybean meal, YC = yellow corn, CGF = corn gluten feed.

    2NDF = neutral detergent fiber.

    3ADF = acid detergent fiber.

    Table 2

    Composition and analyzed nutrient composition of experimental diets, as fed-basis (g/kg)

    g CGF/kg feed

    0

    65

    130

    195

    Yellow corn

    762.9

    716.9

    671.2

    625.6

    Soybean meal

    155.9

    136.7

    117.4

    98

    Corn gluten feed

     

    65.7

    131

    196.4

    Corn oil

    40.0

    40.0

    40.0

    40.0

    Calcium carbonate

    17.9

    19.2

    20.5

    21.9

    Dicalcium phosphate

    10.4

    8.4

    6.3

    4.1

    Salt

    3.5

    3.5

    3.5

    3.5

    L-Lysine HCl

    2.5

    2.4

    2.4

    2.3

    Tryptosine

    1.4

    1.9

    2.4

    2.9

    Threonine

    0.2

       

    Vitamins1

    1.6

    1.6

    1.6

    1.6

    Minerals2

    0.7

    0.7

    0.7

    0.7

    Chromium oxide

    3.0

    3.0

    3.0

    3.0

    Chemical analysis

        

    Dry matter

    904.3

    908.3

    904.9

    907.1

    Protein

    128.7

    127.7

    140.2

    135.7

    Energy, MJ

    17.6

    17.6

    17.6

    17.2

    NDF

    103.5

    121.8

    138.3

    149.2

    Amino acids

        

    Alanine

    7.4

    8.5

    9.8

    9.2

    Arginine

    8.8

    9.8

    11.0

    9.6

    Aspartic acid

    13.5

    14.0

    15.5

    13.3

    Cystine

    2.9

    2.9

    2.7

    2.4

    Glutamic acid

    24.1

    27.1

    31.6

    28.0

    Glycine

    6.6

    7.2

    8.1

    7.4

    Histidine

    3.9

    4.6

    4.9

    5.0

    Isoleucine

    2.6

    4.3

    5.3

    5.0

    Leucine

    9.8

    12.3

    14.4

    13.3

    Lysine

    9.3

    9.8

    9.8

    9.4

    Methionine

    1.8

    1.7

    1.7

    1.4

    Phenylalanine

    5.3

    6.2

    7.1

    6.1

    Proline

    8.3

    11.8

    12.2

    8.4

    Serine

    7.3

    7.6

    8.8

    7.5

    Threonine

    6.4

    6.9

    7.8

    7.1

    Tyrosine

    4.3

    5.2

    5.8

    5.0

    Valine

    3.8

    6.3

    7.4

    7.3

    1Provided per kg piglet diet: Cl, 1.65 g; Na, 0.87 g; Cu, 7.7 mg; Fe, 89.25 mg; Mn, 19.98 mg; Se, 0.087 mg; I, 0.053 mg.

    2Provided per kg piglet diet: vitamin A, 6600 IU; vitamin D, 660 IU; vitamin E, 100 IU; choline, 350 mg; niacin, 54 mg; pantothenic acid, 13.15 mg; riboflavin, 2.2 mg; B12, 36 μg.

    NDF = neutral detergent fiber.

    Sample collection

    The experimental period lasted 12 d; this included 5 d for adaptation to the diet, 5 d for the collection of feces, and 2 d for the collection of ileal digesta. Ileal digesta was collected in plastic bags (length, 11 cm; width, 5 cm) containing 10 mL of 0.2 mol/L solution of HCl to block any bacterial activity. Bags were attached to the barrel of the cannula by using a rubber band. Ileal digesta was collected continuously over the course of 12 h each day. When the bags were filled, they were transferred to a container and frozen at -20°C until lyophilization. All fecal samples were collected, frozen, and kept at -20°C. At the end of the experimental period, the feces were defrosted and homogenized to obtain 10% of the weight as a final sample for lyophilizing.

    Chemical analysis

    Ileal digesta and feces samples were lyophilized and ground in a laboratory mill by using a 0.5-mm mesh (Arthur H. Thomas Co., Philadelphia, PA). Raw materials, experimental diets, ileal digesta, and feces were analyzed for dry matter (DM) and crude protein (CP) according to methods 934.01 and 976.05 of the Association of Official Agricultural Chemists (AOAC) [16]; neutral detergent fiber (NDF), according to van Soest [17]; and energy, by using an oxygen bomb calorimeter (model 1281; Parr, Moline, IL). Chromic oxide levels in the diets, ileal digesta, and feces were determined according to Fenton and Fenton [14]. Amino acid analysis was performed following method 994.12 of the AOAC [16]; samples were hydrolyzed in 6 mol/L HCl at 110°C for 24 h. Methionine and cystine were oxidized with performic acid before the analysis. The amino acid analysis was performed according to the method reported by Henderson et al. [18] by using a high-performance liquid chromatography (HPLC) model (1100; Hewlett Packard).

    Data analysis

    Apparent ileal or total tract digestibility (AID or ATTD) were estimated using the equation proposed by Fan and Sauer [19].
    AID = 1 ID × AF / AD × IF × 100 ,

    where AID% is the apparent (ileal or total) digestibility of a nutrient in the diet, ID is the marker concentration in the diet (mg/kg of DM), AF is the concentration of nutrient in the ileal digesta or feces (mg/kg of DM), AD is the concentration of the nutrient in the diet (mg/kg of DM), and IF is the marker concentration in the ileal digesta or feces (mg/kg of DM).

    The standardized ileal digestibility (SID) was obtained using the formula proposed by Furuya and Kaji [20].
    SID = AID + Endogenous / Dietary Content

    where SID is the standardized ileal digestibility of a nutrient, AID is the coefficient of apparent ileal digestibility of a nutrient, and Endogenous is the endogenous ileal losses of a nutrient in mg/kg of dry matter intake. The calculations were performed using endogenous values reported by Mariscal-Landin and Reis de Souza [21]. Dietary Content is the amount of nutrient consumed in mg/kg of dry matter intake.

    Statistical analysis

    Data were analyzed according to a randomized complete block design [22] by using the general linear model (GLM) procedure of statistical analysis system (SAS) [23]: four blocks with four pigs each (one pig per treatment). Each pig was the experimental unit, and an alpha value of 0.05 was used to assess the significance. The trend of the response (linear or quadratic) was determined using orthogonal contrasts [22]. When a linear effect was determined, a linear equation was obtained using the regression (REG) procedure of SAS [23].

    Results

    Apparent ileal digestibility

    The results of apparent ileal digestibility are shown in Table 3. The inclusion of CGF significantly reduced (P < 0.05) the AID of dry matter; there was a reduction of 5.7 percentage units between the diet without CGF and the diet containing 195 g of CGF (87.7% vs. 82.0%). This adverse effect was also observed in energy digestibility, which was reduced by 4.9 percentage units (from 88.8 to 83.9 in the diets with 0 or 195 g of CGF, respectively). The digestibility of phenylalanine, cystine and proline decreased linearly (P < 0.05) in response to CGF increment in the diet. The average reduction in ileal digestibility of amino acids was 0.031 percentage units per gram of CGF included in the diet. Cystine digestion was the most affected (0.048 percentage units) and phenylalanine, the least (0.011 percentage units). The digestibility of the other amino acids was not affected by the inclusion of CGF at the levels used in this study.
    Table 3

    Apparent ileal digestibility (AID) coefficients (%) of experimental diets

    g CGF/kg feed

    0

    65

    130

    195

    SEM

    Dry matterA

    87.7

    83.8

    82.8

    82.0

    0.66

    Protein

    84.9

    81.4

    81.7

    80.7

    0.98

    EnergyA

    88.8

    86.0

    84.6

    83.9

    0.66

    Amino acids

         

    Alanine

    82.4

    81.1

    80.3

    81.5

    1.31

    Arginine

    93.1

    91.9

    91.0

    89.5

    0.86

    Aspartic acid

    86.1

    83.1

    83.1

    81.5

    1.05

    CystineA

    86.1

    85.1

    81.8

    76.7

    1.22

    Glutamic acid

    88.7

    88.1

    87.7

    86.9

    0.92

    Glycine

    80.2

    79.3

    77.5

    76.3

    1.38

    Histidine

    87.5

    86.7

    85.7

    85.7

    0.91

    Isoleucine

    79.0

    83.5

    83.1

    83.1

    1.63

    Leucine

    87.4

    87.4

    87.4

    86.9

    1.03

    Lysine

    92.7

    90.8

    90.6

    90.6

    0.67

    Methionine

    83.3

    82.4

    82.1

    78.2

    1.20

    PhenylalanineA

    93.7

    92.1

    92.0

    91.4

    0.30

    ProlineA

    87.5

    89.1

    83.9

    79.6

    1.27

    Serine

    85.5

    82.8

    82.7

    81.1

    1.18

    Threonine

    81.9

    77.8

    77.5

    77.2

    1.52

    Tyrosine

    88.2

    88.1

    87.3

    85.9

    0.94

    Valine

    79.8

    85.2

    84.1

    83.5

    1.57

    ALinear effect (P < 0.05).

    Standardized ileal digestibility

    The standardized ileal digestibility of amino acids is shown in Table 4. In general, the inclusion of CGF in the diet did not affect (P > 0.05) the SID. However, a linear decrease in SID of phenylalanine (0.015 percentage units; from 98.7 to 95.8), cystine (0.043 percentage units, from 91.4 to 83.1), and proline (0.047 percentage units, from 103.7 to 94.9) was noted per gram inclusion of CGF. There was no significant reduction in the digestibility of the other amino acids (P > 0.05).
    Table 4

    Standardized ileal digestibility (SID) coefficients (%) of experimental diets

    g CGF/kg feed

    0

    65

    130

    195

    SEM

    Protein

    93.7

    90.3

    89.9

    89.1

    0.98

    Amino acids

         

    Alanine

    87.6

    85.7

    84.3

    85.7

    1.31

    Aspartic acid

    91.2

    88.0

    87.4

    86.6

    1.05

    Arginine

    97.1

    95.5

    94.7

    92.7

    0.86

    CystineA

    91.4

    90.3

    87.3

    83.1

    1.22

    Glutamic acid

    91.9

    91.0

    90.1

    89.7

    0.92

    Glycine

    92.0

    89.9

    86.9

    86.8

    1.38

    Histidine

    91.8

    90.3

    89.2

    89.1

    0.91

    Isoleucine

    90.3

    90.5

    89.0

    88.9

    1.63

    Leucine

    92.5

    92.0

    90.9

    90.6

    1.03

    Lysine

    96.8

    94.2

    94.2

    93.5

    0.67

    Methionine

    87.2

    86.2

    86.6

    83.2

    1.20

    PhenylalanineA

    98.7

    96.5

    95.8

    95.8

    0.30

    ProlineA

    103.7

    100.5

    95.5

    94.9

    1.27

    Serine

    92.4

    89.3

    88.4

    87.9

    1.18

    Threonine

    90.8

    86.0

    85.6

    84.5

    1.52

    Tyrosine

    91.4

    90.7

    89.7

    88.7

    0.94

    Valine

    90.3

    91.6

    89.5

    89.0

    1.57

    ALinear effect (P < 0.05).

    Apparent total tract digestibility

    The ATTD of dry matter, protein, energy, and NDF diminished (P < 0.05) linearly with an increase in CGF concentration in the diet (Table 5). The effect of CGF inclusion on ATTD was net and clear. The reduction of digestibility of dry matter and energy was 6.7 percentage units; for protein, 6.3 percentage units; and for NDF, 11.3 percentage units. The linear equations are shown in Table 6; a decrease of 0.02 to 0.03 percentage units in the ATTD of DM, CP, and energy per gram of CGF included in the diet and a decrease of 0.041 percentage units in the ATTD of NDF were noted per gram of CGF included in the diet.
    Table 5

    Apparent total tract digestibility (ATTD) coefficients (%) of experimental diets

    g CGF/kg feed

    0

    65

    130

    195

    SEM

    Dry matterA

    88.6

    81.3

    83.7

    81.9

    0.36

    ProteinA

    86.9

    76.9

    83.4

    80.6

    0.59

    EnergyA

    88.7

    81.4

    84.1

    82.0

    0.36

    NDFA

    62.5

    47.8

    55.2

    51.2

    1.42

    ALinear effect (P < 0.05).

    NDF = neutral detergent fiber.

    Table 6

    Linear relationships between corn gluten feed (CGF) inclusion and ileal and total tract digestibility coefficients

    Apparent ileal digestibility

      

    Dry matter

    DM = 86.77 (±1.02) – 0.028 (±0.008) gCGF1

    r2 = 0.86

    Energy

    E = 88.22 (±0.96) – 0.025 (±0.008) gCGF

    r2 = 0.92

    Phenylalanine

    Phe = 93.31 (±0.46) – 0.011 (±0.004) gCGF

    r2 = 0.84

    Cystine

    Cys = 87.15 (±1.90) – 0.048 (±0.016) gCGF

    r2 = 0.92

    Proline

    Pro = 89.36 (±1.89) – 0.045 (±0.016) gCGF

    r2 = 0.78

    Standardized ileal digestibility

      

    Phenylalanine

    Phe = 98.13 (±0.51) – 0.015 (±0.004) gCGF

    r2 = 0.78

    Cystine

    Cys = 92.19 (±1.88) – 0.043 (±0.015) gCGF

    r2 = 0.94

    Proline

    Pro = 103.20 (±1.84) – 0.047 (±0.015) gCGF

    r2 = 0.93

    Apparent total tract digestibility

      

    Dry matter

    DM = 86.52 (±1.16) – 0.027 (±0.010) gCGF

    r2 = 0.47

    Protein

    CP = 83.85 (±1.74) – 0.020 (±0.014) gCGF

    r2 = 0.14

    Energy

    E = 86.64 (±1.20) – 0.027 (±0.010) gCGF

    r2 = 0.46

    NDF

    NDF = 58.13 (±2.97) – 0.041 (±0.015) gCGF

    r2 = 0.29

    1gCGF = grams of corn gluten feed added to the diet.

    NDF = neutral detergent fiber.

    Discussion

    Fiber has been defined as “vegetal compounds, from carbohydrate nature and resistant to digestive enzymes”; it is classified according to solubility as soluble fiber (β-glucans, gums, and mucilages) and insoluble fiber (cellulose and most hemicelluloses) [24]. CGF is a mixture of corn structures that remain after the removal of most starch and gluten germ [2]. Approximately, two-thirds of these corn structures are fibrous structures, and one-third consists of soluble compounds [25]. The fiber obtained from corn is essentially insoluble (99% of the total fiber) and represents over 30% of the dry matter [25]; the CGF used in this study had 338.7 g NDF/kg. Insoluble fiber is known to cause endogenous protein losses and reduce the AID of protein and amino acids because it increases mucin secretion as well as mucosal cell shedding via its abrasive effect [8, 26, 27]. The protein content of CGF is not contained in the fibrous part (bran). It is present in the soluble fraction that is mixed with bran at the end of the milling process [2]; therefore, fiber does not block the enzyme access to proteins as is noted in wheat bran, in which a high proportion of protein is present in the aleurone cells [28]. Furthermore, several authors [2931] have suggested that insoluble fiber has a minor effect on the ileal digestibility of amino acids and protein. Both findings (protein is not present in the fibrous part and the minor effect of insoluble fiber on the ileal digestibility of protein) could explain the almost null effect of CGF on the ileal digestibility of protein and amino acids. The decrease in the ileal digestibility of proline could be because of the proteins present in the fibrous fraction; these proteins are rich in extensine—a proline-rich protein that resembles collagen—and are closely linked to the cellulose fraction [3234]. The negative effect on the ileal digestibility of dry matter and energy was due to an increase in the levels of non-digestible compounds (insoluble fiber) after treatment with CGF; as mentioned before, two-thirds of the CGF is insoluble fiber, and hence, it dilutes the digestible dry matter and energy as has been reported previously [35].

    In the large intestine, the undigested food (mainly non-starch polysaccharides and protein) is fermented by microorganisms [36]. This is a long process, but the insoluble fiber decreases the residence time of digesta in the cecum and colon, and this effect is related to the fiber content of the diet [24, 29, 35]. Insoluble fibers are resistant to fermentation; therefore, they play a major role in fecal bulking, unlike soluble fibers that are almost completely fermented and have little effect on increasing fecal bulk [37]. This limited fermentation of insoluble fibers by intestinal bacteria reduces the total digestibility of protein and energy [31, 35, 38]; consequently, the digestible energy is low in diets containing CGF. Furthermore, fibers increase the production of mucin, an almost indigestible protein, thereby increasing protein excretion in the feces [39] and lowering protein digestion. As discussed before, the inclusion of CGF in diets has a mild effect on the ileal digestibility of amino acids; however, it adversely affects energy digestibility. Few studies have quantitatively investigated the adverse effect of NDF on energy digestibility; the findings reported in this study are in agreement with those reported by Dégen [31]. However, in sows, fiber plays an important role in avoiding stereotyped conducts, and thus, sows are able to obtain more energy from fiber than growing pigs [24, 40, 41].

    Conclusions

    CGF did not affect the apparent and standardized ileal digestibility of protein and most amino acids, except that of phenylalanine, cystine, and proline. However, it linearly decreased the ileal digestibility of energy and the total tract digestibility of protein and energy.

    Abbreviations

    CGF: 

    Corn gluten feed

    AID: 

    Apparent ileal digestibility

    SID: 

    Standardized ileal digestibility

    ATTD: 

    Apparent total tract digestibility

    NDF: 

    Neutral detergent fiber.

    Declarations

    Acknowledgments

    This study was funded in part by the Ministry of Agriculture, Livestock and Fishing (SAGARPA) of Mexico and The National Council for Science and Technology (CONACYT) of Mexico via the financial support provided to Research Project SAGARPA-CONACYT 2003-2-169.

    This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

    Authors’ Affiliations

    (1)
    Centro Nacional de Investigación en Fisiología Animal, Instituto Nacional de Investigaciones Forestales Agrícolas y Pecuarias, Km 1, Carretera a Colón, Ajuchitlán Colón, Querétaro, 76280, México
    (2)
    Facultad de Ciencias Naturales, Universidad Autónoma de Querétaro, Santiago de Querétaro, Av. De las Ciencias s/n Juriquilla, 76000 Querétaro, Querétaro, México

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    © Mariscal Landín et al.; licensee BioMed Central Ltd. 2014

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