Livestock Research for Rural Development 28 (10) 2016 Guide for preparation of papers LRRD Newsletter

Citation of this paper

Performance of sheep and goats finished in enriched Caatinga and given supplementation

R F Soares, J M Pereira Filho, K V M F Gama, A M A Silva, M F Cezar, J M Ramos, J B Borburema and O A Bakke

Federal University of Campina Grande, Patos Campus.
Avenida Universitária, Santa Cecília, Patos - Paraíba - Brazil, CEP: 58710-110, P O Box 61.
jucileidebarboza@hotmail.com

Abstract

The present study aimed to assess the performance of sheep and goats finished in thinned Caatinga enriched with Cenchrus ciliares L. that were given supplements. Twelve (12) Santa Inês sheep and 12 F1 crossbred goats (Boer x SRD) with live weight of 24.29 ± 1.35 kg were used in the experiment. The experimental area was 2.4 ha, divided into four plots of 0.6 ha, two for each animal species. For assessment of the vegetation, each plot was subdivided into 2 sub-plots of 0.3 ha. The animals were kept in pasture from 8:00 to 17:00 h, and then were brought into the stalls to receive energy or protein-energy supplementation in an amount equivalent to 1.0% of the body weight. A completely randomized design was used with 2 x 2 factorial arrangement (animal species x type of supplementation).

The finishing of sheep and goats in thinned Caatinga enriched with buffel grass was more favorable to the performance of goats compared to the consumption of dry matter, organic matter, crude protein and neutral detergent fiber, with weights and carcass yields unchanged, though the lambs had better feed conversion. Sheep and goats finished in thinned Caatinga enriched with buffel grass were more favored by protein-energy supplementation.

Keywords: carcass, dry matter intake, pasture, weight gain


Introduction

Most sheep and goats from Northeastern Brazil are raised on an extensive farming system, being mostly fed on native pasture of Caatinga, which is usually improperly handled, exceeding its support capacity, resulting in low performance of the animals and causing significant environmental impact. However, animal production can be considerably increased with the use of techniques for handling woody vegetation, as well as with dietary supplementation.

The control of woody species without forage value in the Caatinga, followed by the introduction of suitable grasses, has become a common practice among producers, whose purpose is to increase the participation of the herbaceous layer in the production of edible dry matter. The Cenchrus ciliaris L. is an exotic grass, resistant to drought and defoliation, which, depending on the physiological stage has protein levels of 3 to 15%. Also, it is one of the species with potential for enrichment of Caatinga, increasing the production and quality of dry matter, improving the support capacity and the performance of goats and lambs, whose meat is the main source of proteins for the population of the northeastern semiarid region. Yet, most producers still have low production rates.

Santa Inês sheep and F1 Boer x SRD goats are characterized by their capacity for adaptation to the semiarid region, being resistant to diseases, and represent an important source of income for farmers in the Northeast of Brazil. Quinzeiro Neto et al (2011) stressed that sheep has great potential for meat production. Maia et al (2012) reported that, because of their higher growth rate and good carcass yield, Santa Inês sheep are becoming more popular among producers compared to other woolless sheep.

Boer goats are characterize by: early growth, good conformation; good rates of fertility, fecundity, prolificacy and longevity; excellent maternal qualities with adequate milk production and high rates of weaning (Marques et al 2013), being very suitable for crossing with SRD animals and native breeds. The crossing of Boer goats with SRD animals produces offspring with higher growth rate (Freitas et al 2011). However, these animals do not demonstrate their entire production potential because of their breeding system, particularly due to the decrease in the quality and quantity of dry matter available throughout the year. Thus, the Caatinga vegetation becomes scarce, which requires an improved management, with the introduction of some sort of supplementation.

Few studies use supplementation for goats and sheep under Caatinga conditions. On the other hand, there are doubts about the supplementation that should be used, once the amount and quality of pasture the animals are put to graze may require different types of supplementation. Carvalho Júnior et al (2011) reported that the use of concentrates in the diet of ruminants can increase the intake of energy and protein, and, thus, meet the demand of animals with higher productivity.

Silva et al (2010) stressed that ruminants that graze in semiarid regions, including under Caatinga conditions (Leite 2002) usually require more energy than protein. However, the actual need for supplementation will be determined by the botanical and chemical composition of the pasture and diet of the animals, which is decisive to obtain animals with better performance and carcass yield compatible with the consumer market. Thus, the present study aimed to assess the performance of sheep and goats finished in thinned Caatinga enriched with Cenchrus ciliaris L. cv. Biloela that were given two types of supplementation.


Material and methods

Experimental area

The study was conducted at Farm Lameirão of Federal University of Campina Grande, located at the coordinates 7º1’ South latitude and 35º1’ West longitude. According to Köppen’s classification, the region has a BShw climate – semiarid, with short rainy seasons in summer-autumn and rainfall mainly concentrated in the months of March and April. The annual rainfall may vary from 150 to 1300 mm, but the highest average precipitation ever recorded is 500 mm. In 2010, year in which the experiment was conducted, the monthly rainfall from January to December was respectively: 117.7; 31.8; 70.9; 188.2; 11.3; 61.5; 2.1; 0.0; 0.0; 70.9; 0.0 and 158.6 mm, totaling 713 mm: The dry period varies from six to eight months, beginning in June and ending in mid-January. The average annual temperature is around 28 ºC, with maximum and minimum temperatures of approximately 35 and 22 ºC, respectively. The average relative humidity is 60%.

The woody vegetation of the experimental area was submitted to selective thinning to allow 15% of soil coverage (Araújo Filho 2013), which consisted in the partial removal of undesirable species, preservation of species with high timber value and plants considered endangered and/or that stay green the year round. For the control of thinned species, regrowths were cut during the rainy season. Caatinga was enriched with Cenchrus ciliaris L. cv. biloela soon after thinning, with the seeds sown by scattering, and distributed as evenly as possible.

Animals and diets

The experiment was submitted to analysis and approved by the Research Ethics Committee of Universidade Federal de Campina Grande - Campus de Patos (Protocol 29/2008).

Twelve (12) Santa Inês sheep and 12 crossbred goats with live weight of 24.29 ± 2.38 kg were used in the experiment. The animals were individually identified with numbered collars. The experimental area was 2.4 ha, divided into four plots of 0.6 ha, two for each animal species, all of them with shelter and water fountain. For assessment of the vegetation, each plot was divided into two subplots of 0.3 ha. During the experiment, the animals received the routine treatments, such as vaccinations and endoparasite and ectoparasite control. The animals were randomized, with six animals per plot (sheep or goats), corresponding go the continuous grazing stocking number of 0.54 UA/ha.

When the availability of DM of the herbaceous stratum surpassed 2000 kg/ha, or else, more than 50% of the potential of the thinned Caatinga (Araújo Filho 2013), two plots were occupied with sheep and two with goats. The diet was composed of native pasture enriched with Cenchrus ciliaris and supplementation. The animals were kept in pasture from 8:00 to 17:00 h, and then were brought into the stalls to the energy supplement (ES) based on maize meal (970 g/kg) or the protein-energy supplement (PES), consisting of soybean meal (500 g/kg) and ground maize grain (470 g/kg). Both supplements contained 30 g/kg of mineral premix for sheep (Santa Inês) or goat for the F1 Boer x SRD goats. The supplementation was provided in an amount equivalent to 10 g/kg of the body weight that was weekly adjusted. (NRC 2007). The animals received water ad libitum in the field and in the stalls. The chemical composition of the supplements is described in Table 1.

Table 1. Chemical composition of the supplement provided to the sheep and goats

Component

Type of Supplementation

Energy Supplement

Protein-Energy Supplement

Dry Matter1

901

903

Mineral Matter2

16

38

Organic Matter2

984

962

Crude Protein2

118

269

Neutral Detergent Fiber2

235

244

Acid Detergent Fiber2

48

80

¹g/kg of NM (natural matter); 2g/kg of DM.

Sampling and chemical analyses

For estimating consumption, the combination of estimated fecal output, which was determined based on the external indicator hydroxyphenyl propane (LIPE ®) recommended by Rodríguez et al (2007) with the in vitro digestibility of the material collected in the rumen. After 30 days of the experiment, the indicator was daily administered at a dosage of a 250 mg capsule directly into the rumen, through a tube, during 5 days. From the third day of administration of the LIPE® samples of feces were manually collected directed from the rectal ampulla of the animals, homogenized to form animal samples, which were pre-dried, milled and stored in bottles sent to the Department of Chemistry of Instituto de Ciências Exatas of UFMG for estimates of fecal production.

The concentration of LIPE® was determined by red spectrometer (Rodríguez et al 2007). The estimated production of feces (FP) was obtained by the equation: FP={(total LIPE administered/concentration of LIPE in the feces) x 100}.

The total collection in the rumen was made in two sheep and two goats with a ruminal fistula. On the day before collection, the animals were brought into the stalls, fasted for 16 hours and then the total ruminal content was collected. The animals were allowed to graze for 20 minutes and then all the ruminal material was collected and stored in polystyrene bottles with ice for further determination of dry matter (DM), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF) and crude energy. For analysis of in vitro dry matter digestibility (IVDMD), 0.500 g of the sample were weighed in TNT bags and then incubated in ANKOM 200 device (Ankon Technology Corp., Faiport, NY, USA), according to the methodology described by Silva and Queiroz (2002).

Slaughter and measurements

When the animals reached 35 kg of live weight or 75 days of experiment, they were fasted for 24 hours (solid) 16 hours (liquid), and weighed to obtain the slaughter weight (SW). At slaughter, stunning and bleeding (both jugular veins and carotid arteries severed), followed by skinning and evisceration. The gastrointestinal tract, bladder and gall bladder were removed, weighed and emptied to obtain the weights of the contents, and, thus, the empty body weight (EBW), which was calculated by subtracting the weights of the contents from the SW.

The carcass was obtained after removal of the head and paws, and the hot carcass weight (HCW) was obtained. All components of the animal’s body not included in the carcass weight were called “non-carcass components”, which were obtained by subtracting the HCW from the EBW. Among the non-components, the weight and yield of liver, kidneys and gastrointestinal tract, tongue, esophagus, trachea and lungs, heart, head and paws were assessed. With HCW the yields of the hot carcass (HCY) and biological yield (BY) were estimated, and then the carcasses were cooled at 4 ºC for 24 hours to obtain the cold carcass weight (CCW) and the amount of carcass loss by cooling shrinkage (CS), with the methodology described by Cezar and Sousa (2007).

Statistical analyses

A completely randomized design was used with 2 x 2 factorial arrangement, with two animal species (sheep and goat) and two types of supplementation (energy and protein-energy). The data were subjected to analysis of variance and the means were compared by Tukey test. The data were analyzed at 5% probability level and processed by SAS (Statistical Analysis System, Version 9.1, 2003).


Results

Chemical composition, intake and digestibility

The chemical composition of the vegetation did not change in the areas where the sheep and goats grazed, reflecting the experimental planning adopted (Table 2). Dicotyledons had a crude protein (CP) content of 106 g/kg, and dry matter (DM) content of 103 g/kg.

It is important to stress that the values of NDF of dicotyledons were 596 g/kg and 619 g/kg in the areas grazed by sheep and goats, respectively.

Table 2. Chemical composition of herbaceous vegetation of thinned Caatinga enriched with Cenchrus ciliaris L. grazed by sheep and goats

Item

DM¹

MM²

OM²

CP²

NDF²

ADF²

Buffel grass

Sheep grazing

532A

82A

918A

47A

769A

494A

Goat grazing

538A

74A

927A

51A

784A

504A

SEM

2.84

1.19

1.19

0.53

2.25

2.62

p

0.67

0.20

0.20

0.28

0.24

0.50

Dicotyledons

Sheep grazing

440A

63A

940A

106A

596A

460A

Goat grazing

461A

60A

937A

103A

619A

484A

SEM

9.54

0.82

0.82

2.12

5.47

3.83

p

0.67

0.51

0.51

0.75

0.44

0.26

Other grasses

Sheep grazing

529A

67A

933A

49A

766A

474A

Goat grazing

512A

65A

935A

59A

756A

471A

SEM

3.91

1.64

1.64

1.35

3.28

2.80

p

0.41

0.81

0.81

0.16

0.53

0.83

DM = dry matter; MM = mineral matter; OM = organic matter; CP = crude protein; NDF = neutral detergent fiber; ADF = acid detergent fiber. Means with different letters in the column of the vegetable component differ (P<0.05) by Tukey test. SEM = standard error; p = probability; ¹g/kg of NM; 2g/kg of DM.

The DM content of Cenchrus ciliaris L. was lower in the May-July period than in the other periods, which can be associated to the phenological cycle of the plant (Table 3). The highest DM content was observed in August, after the rainfall that lasted until July.

The CP content of Cenchrus ciliaris L., even in May 5, when DM content was 444 g/kg of NM (natural matter) was considered low (52 g/kg of DM). The increase in CP content (65 g/kg) in July 5, em reflected the occurrence of rainfall, which made the regrowth of Cenchrus ciliaris L possible and, thus, its selection by the animals.

Table 3. Chemical composition of herbaceous vegetation in a thinned Caatinga enriched with Cenchrus ciliaris L. and grazed by sheep and goats at different assessment times

Item

DM¹

MM²

OM²

CP²

NDF²

ADF²

Buffel grass

05/May

444C

69.50A

931A

51.70B

790A

512A

05/June

579B

78.80A

921A

39.60A

778A

495A

05/July

444C

82.20A

920A

65.30A

766A

497A

05/August

673A

80.30A

918A

39.10B

772A

493A

SEM

2.84

1.19

1.19

0.53

2.25

2.62

p

0.001

0.49

0.49

0.00

0.53

0.72

Dicotyledons

05/May

303A

57.90A

942A

119A

578A

460A

05/June

473A

61.70A

938A

94.10A

620A

495A

05/July

515A

65.10A

935A

115A

600A

462A

05/August

512A

62.10A

938A

90.30A

632A

473A

SEM

9.54

0.82

0.82

2.12

5.47

3.83

p

0.05

0.68

0.68

0.23

0.57

0.58

Other grasses

05/May

312C

78.10A

922A

74.90A

712A

437A

05/June

534B

79.50A

940A

42.80AB

769A

479A

05/ July

523B

79.50A

921A

33.30B

786A

486A

05/August

712A

44.90A

955A

64.30AB

777A

486A

SEM

3.91

1.64

1.64

1.35

3.28

2.80

p

0.001

0.07

0.07

0.01

0.06

0.12

DM = dry matter; MM = mineral matter; OM = organic matter; CP = crude protein; NDF = neutral detergent fiber; ADF = acid detergent fiber. Means with different letters in the column within the vegetable component differ (P<0.05) by Tukey test. SEM = standard error; P = probability; ¹g/kg of NM; 2g/kg of DM.

There were no leftovers of the supplement offered to the animals, and the intake of concentrate (DM and OM) by the animals (10g of DM/kg of LW). The goats consumed more DM and OM of bulk and total (g e g/kg of MW) than sheep (Table 4).

Regardless of the type of supplementation, the intake of DM and OM by the animals (Table 4) was similar, reflecting the high selective capacity of these animals.

Table 4. Dry matter and organic matter intake depending on the animal species and type of supplementation

Variable

Animal species

p

Type of supplementation

p

SEM

Sheep

Goats

Energy

Protein-energy

Dry matter intake

Concentrate (g)

263

282

0.35

283

262

0.35

37.18

Bulk (g)

412

561

0.02

505

468

0.52

94.99

Total (g)

675

843

0.03

788

730

0.41

116.64

Concentrate (g/kgPV)

10

10

-

10

10

-

-

Bulk (g/kgPV)

16

20

0.05

18

18

0.82

2.94

Total (g/kgPV)

26

30

0.05

28

28

0.82

2.94

Concentrate (g/kgPM)

23

23

0.37

23

23

0.33

0.78

Bulk(g/kgPM)

36

46

0.03

41

41

0.97

6.61

Total (g/kgPM)

58

69

0.02

64

63

0.87

6.60

Organic matter intake

Concentrate (g)

256

274

0.41

277

253

0.27

36.13

Bulk (g)

377

519

0.02

465

431

0.52

87.81

Total (g)

634

793

0.03

742

684

0.38

109.13

Concentrate (g/kgPV)

10

10

-

10

10

-

-

Bulk (g/kgPV)

15

18

0.04

16

17

0.83

2.72

Total (g/kgPV)

24

28

0.04

26

26

0.90

2.72

Concentrate (g/kgPM)

22

22

0.37

23

22

0.10

0.76

Bulk (g/kgPM)

33

42

0.02

38

38

0.96

6.11

Total (g/kgPM)

55

65

0.02

60

59

0.79

6.10

p = probability; SEM = standard error; LW = live weight; MW = metabolic weight.

The intake of CP of the concentrate did not differ between the species (Table 5). However, there was difference in the intake of CP (bulk and total), with greater consumption of goats compared to sheep.

Regarding the type of supplementation, there was a difference in the consumption of CP of the concentrate and total.

Table 5. Consumption (intake) of CP and NDF, depending on the animal species and type of supplementation

Variable

Animal species

p

Type of supplementation

p

SEM

Sheep

Goats

Energy

Protein-energy

Consumption of CP

Concentrate (g)

45

49

0.34

25

69

<.00

7.60

Bulk (g)

47

74

0.00

63

58

0.51

12.41

Total (g)

91

123

0.00

88

127

0.00

15.72

Consumption of NDF

Concentrate (g)

31

34

0.40

33

32

0.61

4.46

Bulk (g)

300

407

0.02

367

340

0.52

68.92

Total (g)

331

441

0.02

400

372

0.51

71.02

p = probability; SEM = standard error.

There was no difference between species for digestibility of DM, OM, CP and NDF (Table 6). However, regarding food conversion, the sheep showed 5.19 differing from the 6.02 obtained by the goats. Despite the greater consumption by goats, the performance of the animals did not differ.

Table 6. Digestibility of DM, OM, CP and NDF depending on the animal species and type of supplementation

Digestibility (g/kg)

Animal species

p

Type of supplementation

p

SEM

Sheep

Goats

Energy

Protein-energy

Dry matter

637

640

0.85

642

635

0.71

0.03

Organic matter

666

674

0.63

676

664

0.49

0.03

Crude protein

614

659

0.05

561

712

0.001

0.04

Neutral detergent fiber

547

546

0.97

550

543

0.73

0.03

Food conversion

5.19

6.02

0.001

6.56

4.87

0.14

0.844

Initial weight (kg)

24.31

24.28

-

23.82

24.76

-

2.47

Total weight gain (kg)

8.12

8.57

0.53

7.50

9.18

0.02

1.75

Average daily weight gain (g)

130

140

0.43

120

150

0.05

0.03

p = probability; SEM = standard error.

As for supplementation, only digestibility of crude protein results differs, with a higher value for protein-energy supplementation.

The protein-energy supplementation supported a feed conversion of 4.87, while animals supplemented only with energy had a conversion of 6.56.

Performance and carcass weight

As for performance, there was no difference between the species for total weight gain and average daily weight gain (Table 6). Regarding the types of supplementation, the protein-energy supplementation provided greater total weight gain for the animals.

There was interaction between species and type of supplementation for the final weight (Table 7), with the sheep that received protein-energy supplementation obtaining a higher final weight than the sheep that received energy supplementation.

On the other hand, the final weight of goats was similar in the two types of supplementation (Table 7).

Table 7. Final weight (kg) of sheep and goats finished in thinned Caatinga enriched with Cenchrus ciliaris L. in each type of supplementation

Species

Type of supplementation

p

SEM

Energy

Protein-energy

Sheep

30.22

34.63

0.03

1.99

Goat

32.43

33.25

p = probability; SEM = standard error.

There was no difference between the animal species for slaughter weight, amount of carcass loss by cooling shrinkage and biological yield. As for the slaughter hot carcass, cold carcass and empty body weights of the animals that received protein-energy supplementation, they were greater than those obtained by the animals given energy supplementation (Table 8), indicating the role of the protein of the concentrate in improving the low protein content of the pasture.

Table 8. Characteristics of the carcass of sheep and goats depending on the animal species and type of supplementation

Item

Animal species

p

Type of supplementation

p

SEM

Sheep

Goats

Energy

Protein-energy

Weights (kg)

Slaughter

30

30

0.51

29

31

0.00

1.67

Hot carcass

14

14

0.66

13

14

0.01

1.23

Cold carcass

13

13

0.53

13

14

0.01

1.11

Empty body weight

24

24

0.65

23

25

0.00

1.92

Yield (%)

Biological

58

58

0.96

59

58

0.42

1.90

Hot carcass

46

46

0.95

46

46

0.92

2.37

Cold carcass

45

45

0.77

45

45

0.74

2.27

Carcass loss by cooling shrinkage

4

3

0.15

4

3

0.36

1.32

p = probability; SEM = standard error.


Discussion

Chemical composition, intake and digestibility

In the present study, dicotyledons had a crude protein (CP) content and dry matter (DM) content higher than the minimum dry matter (DM) content of 70 g/kg for microbial growth in the rumen (Van Soest 1994).

The values of NDF of dicotyledons corroborating Formiga et al (2011) who reported that herbaceous dicotyledons tend to have lower NDF than grasses. This aspect is stressed by the contribution of leguminous plants with high protein value among dicotyledons (Carvalho and Pires 2008).

The highest DM content was observed in August, after the rainfall that lasted until July, and which, according to Araújo Filho et al (2002) may induce or hasten the physiological maturation of the plants, and, consequently, increase DM concentration.

The CP content of Cenchrus ciliaris L., even in May 5, when DM content was considered low, from a nutritional view, being lower than the minimum 70.0 g content needed for proper functioning of the rumen (Van Soest 1994).

In the present experiment, the goats consumed more DM and OM of bulk and total than sheep. The grazing habit of goats may have favored the higher intake of bulk, since some herbaceous plants were in a more advanced phonological growth stage and with high altitude, which may difficult intake by sheep. (Osoro et al 2013). The selective capacity of these animals was reported by Pereira Filho et al (2013) who stressed the high degree of use of the Caatinga vegetation by sheep and goats, being considered intermediate selectors with high feeding flexibility, depending on the time of the year, availability and quality of grazing.

The intake of CP (bulk and total), with greater consumption of goats compared to sheep. This finding can be associated to the ability of these animals to select from among herbaceous plants, the broadleaf species, and, among these, the leguminous plants (Celaya et al 2007), which usually have greater protein concentration than grasses, which are usually selected by sheep (Rutter 2006).

Regarding the type of supplementation, a difference in the consumption of CP of the concentrate and total is explained by the composition of supplements with greater amount of CP in the case of protein-energy supplementation. In turn, the similar consumption of NDF in the two types of supplementation reflects the animals’ capacity of selecting their diet, by adjusting the fibrous portion of the diet (Animut et al 2005).

The results for food conversion of sheep, which showed greater efficiency of feed conversion than goats, are consistent.

The results obtained in this paper for digestibility of MM for sheep and goats was 630 g for sheep and 640 g for goats, being consistent with the findings of Formiga et al (2011) who assessed goats and sheep grazing in a Caatinga area enriched with buffel grass supplemented only with minerals, and obtained digestibility values for OM of 0.64 and 0.62 for sheep and goats, respectively. These results are considered satisfactory by the authors because they concern a native pasture at a dry season, confirming the ability of sheep and goats of selecting diets of good nutritious value.

It is important to stress that the greatest intake of CP obtained with the protein-energy supplementation was not sufficient to establish a difference in food conversion.

Performance and carcass weight

In a study with SRD goats, Voltolini et al (2009) found that energy supplementations had no effect on the average daily weight gain and total weight gain.

In a study with Santa Inês sheep finished on pasture that were given different levels of supplementation, Dantas et al (2008) obtained a daily gain of 148 g/day for sheep supplemented with 1% of LW. Carvalho Júnior et al (2011) assessing the performance of crossbred F1 (Boer x SRD) goats finished on native pasture that were given different levels of supplementation obtained a gain of 147 g/day also with 1% of the LW in supplementation.

The result of the interaction between species and type of supplementation to the final weight, indicates that the protein concentration of the protein-energy supplementation must have remedied the deficiency of the pasture, particularly if the CP values of Cenchrus ciliaris L and of the native grasses are considered, which despite the high availability of DM, did not allow the sheep to select a diet that compensated the lower protein intake in the energy concentrate.

Regarding the final weight of goats, the animals managed to supply their needs in the pasture, according to Formiga et al (2011), goats prefer shrubby, broadleaf vegetable species, which have a higher CP content than grasses.

The results for carcasses of this study corroborate the findings of Silva et al (2010), who affirmed that animals in pastures in semiarid conditions need more protein in their diet than the standard level of animals bred in temperate regions or in confinement.


Conclusions


Acknowledgments

The authors acknowledge the financial support by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).


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Received 7 August 2016; Accepted 29 August 2016; Published 1 October 2016

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