Different types of animal bones including fish bones (A), country hen bones (B), broiler hen bones (C), goat trotters (D), goat horns (E), cow bones (F) and buffalo horns (G) were subjected for the chemical tests and instrumental techniques such as basicity test, qualitative analysis of element and quantitative analysis of Ca+ 2. The instrumental techniques which are used are furnace for calcination, atomic absorption spectrophotometry (AAS) and Fourier transform infrared spectroscopy (FTIR). All the samples were also analyzed for microbial activities. In addition, the samples were also tested as nano-biofertilizers. All bones were calcinated at 1000°C. By calcination all bones were converted into the oxide particularly CaO and other oxides. After calcination, the animal bones surface was very smooth, and they contained tiny pore size (nanosized) particles.
At low temperature like 600 oC only organic components are removed and carbonates are not converted into CaO. At high temperatures like 1000 oC or 1200 oC, carbonates are converted into CaO. FTIR spectra of all samples were determined by Fourier transform infrared spectroscopy. Their FTIR spectra showed only one peak at 1050 cm− 1 which is the characteristic peak for CaO (Fig. 3). It was further supported by the reported literature (Engin et al. 2006; Figueiredo et al. 2010).
Buffalo horns showed the highest basicity among the above mentioned bones which means it contains high percentages of Ca, P and N. Goat trotters and goat horns also showed the high basicity but lower than buffalo horns (Table 1). Cow bones and chicken bones (country) showed the lowest basicity while fish bones and chicken bone (broiler) showed the moderate basicity (Table 1). White precipitates were appeared as a result of calcium test which confirmed the presence of calcium in bones and gelatin like white precipitates were appeared as a result of magnesium test which confirmed the presence of magnesium in bones. Buffalo horns showed the highest percentage of Ca+ 2 than all the other bones (Table 2). Cow bones also showed the high percentage of Ca+ 2 but buffalo bones dominated all bones. Similarly, fish bones and chicken bones (country) showed the moderate percentages while chicken bones (broiler), goat trotters and goat horns showed the lowest percentages of calcium. Chicken bones (broiler) contained the lowest percentage of calcium (Table 2). The animal bones were analyzed by flame atomic absorption spectrophotometry which showed the presence of different metals like Ca, Cd, Cu, Fe, K, Mg, Ni, As, Pb etc. The highest percentage of Ca (106.80 mg/l), Mg (2.038 mg/l) and As (9,690 mg/l) were observed in chicken bones (broiler) and the lowest percentage of Ca (89.44 mg/l), Mg (1.656 mg/l) and As (10.30 mg/l) were observed in buffalo horns (Table 3).
Table 1
Determination of basicity of different calcinated bones by chemical method
Name of bones
|
Basicity (m. moles of benzoic acid/g of bones)
|
Fish bones (A)
|
0.025
|
Chicken bones (country) (B)
|
0.015
|
Chicken bones (broiler) (C)
|
0.02
|
Goat trotters (D)
|
0.035
|
Goat horns (E)
|
0.03
|
Cow bones (F)
|
0.015
|
Buffalo horns (G)
|
0.13
|
In the present study, fish bones, chicken bones (both country and broiler), and bones and horns of goats, cows, and buffaloes were collected from fish harbor, poultry markets, and slaughterhouses at different locations in Karachi. To determine their importance as biofertilizers, their macronutrients (C, N, Ca, P, K, and Mg) were evaluated and fertility was assessed by growing kidney bean plants. Plant growth with each fertilizer was checked after 1–5 weeks. Biochar was prepared from different bones/horns, and the filtration capacity of each type of biochar was tested with different heavy metal solutions at different concentrations.
Table 2
Determination of percentage of calcium in different calcinated bones by chemical method
Name of bones and code
|
Ca+ 2 (%)
|
Fish bones (A)
|
53.57
|
Chicken bones (country) (B)
|
52.50
|
Chicken bones (broiler) (C)
|
40.0
|
Goat trotters (D)
|
47.12
|
Goat horns (E)
|
43.19
|
Cow bones (F)
|
57.12
|
Buffalo horns (G)
|
63.90
|
Table 3
Determination of metals content (ppm) in samples by flame Atomic Absorption Spectrophotometer (FAAS).
Sample ID
|
Ca
Mean ± SD
|
Cd
Mean ± SD
|
Cu
Mean ± SD
|
Fe
Mean ± SD
|
K
Mean ± SD
|
Mg
Mean ± SD
|
Ni
Mean ± SD
|
As
Mean ± SD
|
Pb
Mean ± SD
|
1
|
92.73 ± 1.041
|
0.012 ± 0.0001
|
0.406 ± 0.0034
|
3.636 ± 0.0313
|
18.23 ± 0.004
|
5.668 ± 0.0181
|
0.323 ± 0.0012
|
7.786 ± 0.3837
|
0.804 ± 0.0188
|
2
|
98.78 ± 6.187
|
0.013 ± 0.0001
|
0.159 ± 0.0027
|
1.464 ± 0.0135
|
18.28 ± 0.016
|
5.544 ± 0.0199
|
0.301 ± 0.0015
|
8.775 ± 0.5173
|
0.776 ± 0.0102
|
3
|
106.80 ± 8.28
|
0.013 ± 0.0002
|
0.137 ± 0.0964
|
0.396 ± 0.0039
|
18.30 ± 0.000
|
2.038 ± 0.0093
|
0.287 ± 0.0056
|
9.690 ± 0.8197
|
0.724 ± 0.0135
|
4
|
92.95 ± 0.576
|
0.012 ± 0.0002
|
0.159 ± 0.0015
|
5.089 ± 0.0119
|
18.36 ± 0.001
|
1.936 ± 0.0058
|
0.328 ± 0.0027
|
10.88 ± 0.623
|
0.861 ± 0.0242
|
5
|
94.23 ± 1.703
|
0.012 ± 0.0002
|
0.172 ± 0.0034
|
1.682 ± 0.0185
|
18.36 ± 0.001
|
1.960 ± 0.0057
|
0.286 ± 0.0041
|
12.11 ± 0.258
|
0.836 ± 0.0313
|
6
|
96.67 ± 1.908
|
0.012 ± 0.0003
|
0.120 ± 0.0028
|
0.843 ± 0.0060
|
18.37 ± 0.002
|
1.950 ± 0.0066
|
0.305 ± 0.0025
|
13.22 ± 0.346
|
0.930 ± 0.0017
|
7
|
89.44 ± 0.146
|
0.004 ± 0.0002
|
0.924 ± 0.0039
|
2.185 ± 0.0056
|
5.816 ± 0.0191
|
1.656 ± 0.0031
|
0.116 ± 0.0083
|
10.30 ± 0.293
|
0.422 ± 0.0102
|
Total carbon (TC) and total nitrogen (TN) were determined at different temperatures (200°C, 400°C, 600°C, 800°C, and 1000°C) from different samples; fish bones, chicken bones (both country and broiler), goat bones, goat horns, cow bones, cow horns, buffalo bones, and buffalo horns (Table 4). TC and TN tended to decrease with increasing temperature. At a given temperature (200–1000°C), among the different bones and horns samples, broiler chicken bones generally had the lowest TC and TN values (TC = 33 − 1.1 g/Kg; TN = 5.5 − 0.24 g/Kg), while the highest TC and TN values were found in buffalo bone samples (TC = 329 − 14.3 g/Kg; TN = 44-1.21 g/Kg). When comparing bones and horns, higher TC and TN values were found in the bone samples than in the horn samples. During combustion, the concentration of TC and TN was highest at 200°C, while the lowest concentration was measured at 1000°C. At 1000°C, the maximum loss of TC and TN was found to be between 95.5 and 97.5%.
Table 4
Determination of carbon and nitrogen by CHN analyzer
|
|
Total Carbon (g/Kg) at different temp.
|
|
|
Total Nitrogen (g/Kg) at different temp.
|
Samples
|
200°C
|
400°C
|
600°C
|
800°C
|
1000°C
|
200°C
|
400°C
|
600°C
|
800°C
|
1000°C
|
|
Mean ± SD
|
Mean ± SD
|
Mean ± SD
|
Mean ± SD
|
Mean ± SD
|
Mean ± SD
|
Mean ± SD
|
Mean ± SD
|
Mean ± SD
|
Mean ± SD
|
Fish bone
|
125 ± 5.28
|
42.5 ± 2.33
|
7.3 ± 0.75
|
5.5 ± 0.63
|
3.7 ± 0.70
|
19 ± 0.71
|
6.9 ± 0.34
|
2.6 ± 0.26
|
0.8 ± 0.07
|
0.68 ± 0.03
|
Chicken bone (country)
|
112 ± 5.32
|
37.2 ± 1.10
|
6.1 ± 0.50
|
4.7 ± 0.25
|
3.1 ± 0.36
|
15 ± 0.51
|
4.8 ± 0.28
|
1.9 ± 0.22
|
0.6 ± 0.07
|
0.52 ± 0.03
|
Chicken bone (broiler)
|
33 ± 1.47
|
14 ± 0.80
|
3 ± 0.34
|
2.1 ± 0.27
|
1.1 ± 0.18
|
5.5 ± 0.27
|
2 ± 0.23
|
1.1 ± 0.14
|
0.39 ± 0.04
|
0.24 ± 0.02
|
Goat bone
|
317 ± 10.20
|
103.6 ± 4.15
|
21.4 ± 1.06
|
17.4 ± 0.48
|
13.1 ± 0.41
|
41 ± 1.33
|
17.9 ± 0.57
|
3.9 ± 0.31
|
1.3 ± 0.17
|
1.09 ± 0.08
|
Goat horns
|
288 ± 8.62
|
96.1 ± 4.01
|
19.3 ± 0.63
|
15.7 ± 0.42
|
11.4 ± 0.50
|
33 ± 1.36
|
16.1 ± 0.56
|
3.3 ± 0.42
|
1.09 ± 0.08
|
0.99 ± 0.06
|
Cow bone
|
155 ± 7.07
|
52.3 ± 1.88
|
10.5 ± 0.55
|
8.1 ± 0.37
|
6.9 ± 0.38
|
22 ± 0.88
|
10.9 ± 0.51
|
2.7 ± 0.24
|
0.98 ± 0.04
|
0.81 ± 0.04
|
Cow horns
|
122 ± 2.20
|
41.9 ± 0.67
|
8.5 ± 0.43
|
7.7 ± 0.35
|
6 ± 0.34
|
17 ± 0.79
|
8.3 ± 0.31
|
2.1 ± 0.24
|
0.88 ± 0.05
|
0.72 ± 0.03
|
Buffalo bone
|
329 ± 4.86
|
110.2 ± 2.65
|
22.8 ± 1.11
|
18.1 ± 0.35
|
14.3 ± 0.40
|
44 ± 1.08
|
18.1 ± 0.48
|
4.1 ± 0.36
|
1.42 ± 0.14
|
1.21 ± 0.03
|
Buffalo horns
|
299 ± 5.24
|
98.1 ± 2.49
|
19.4 ± 0.86
|
17.2 ± 0.44
|
13.2 ± 0.48
|
37 ± 1.41
|
17.5 ± 0.46
|
3.5 ± 0.45
|
1.22 ± 0.07
|
1.1 ± 0.03
|
The total content of calcium (Ca), phosphorus (P), potassium (K) and magnesium (Mg) in the calcified bone samples was determined. Overall, among the different samples, the lowest concentration of these four elements was recorded in the bones of broiler chickens (Ca; 55 g/Kg, P; 26 g/Kg, K; 2.5 g/Kg and Mg; 0.83 g/Kg), while the highest concentrations were recorded in the bones of buffaloes (Ca; 112 g/Kg, P; 54 g/Kg, K; 5.5 g/Kg and Mg; 1.9 g/Kg). When comparing bone and horn samples, the concentration of these elements was higher in the bones than in the horn samples from the same animal. The concentration in the bone country chickens was higher than in those of broiler chickens (Table 5).
Table 5
Determination of Ca, K and Mg by atomic absorption spectroscopy and P by UV-VIS spectroscopy
Samples
|
Ca
Mean ± SD
|
P
Mean ± SD
|
K
Mean ± SD
|
Mg
Mean ± SD
|
Fish bone
|
95 ± 3.62
|
46 ± 1.21
|
5 ± 0.45
|
1.8 ± 0.09
|
Chicken bone (country)
|
81 ± 1.83
|
40 ± 0.89
|
4.2 ± 0.34
|
1.4 ± 0.18
|
Chicken bone (broiler)
|
55 ± 1.05
|
26 ± 0.75
|
2.5 ± 0.12
|
0.83 ± 0.01
|
Goat bone
|
107 ± 2.88
|
52 ± 1.33
|
5.3 ± 0.44
|
1.8 ± 0.21
|
Goat horns
|
99 ± 2.59
|
47 ± 1.51
|
4.5 ± 0.39
|
1.5 ± 0.14
|
Cow bone
|
101 ± 2.32
|
48 ± 1.69
|
5 ± 0.52
|
1.8 ± 0.40
|
Cow horns
|
96 ± 2.64
|
47 ± 1.38
|
4.8 ± 0.36
|
1.7 ± 0.21
|
Buffalo bone
|
112 ± 3.39
|
54 ± 1.76
|
5.5 ± 0.38
|
1.9 ± 0.23
|
Buffalo horns
|
105 ± 3.16
|
51 ± 1.97
|
5.2 ± 0.51
|
1.7 ± 0.27
|
To determine the fertility effect of the biofertilizers prepared from the bone/horn samples, kidney bean plants were grown in pots containing the biofertilizers prepared from the bone samples. Seeds were sown in each pot and their growth was observed for up to 5 weeks. Each week, the size of the stems and leaves of each plant was determined (cm). The plant grown in buffalo bone biofertilizer showed maximum growth after 5 weeks (leaves 7.5 cm and stems 27.2 cm), while the plant grown in the broiler chicken bone biofertilizer showed minimum growth (stems 15.2 cm, leaves 4 cm). In comparison, the plants fertilized with the horns grew less than the plants fertilized with the bones of the same animal (Table 6).
Table 6: Determination of biofertilizer efficacy of different samples
S. No.
|
Name of Samples
|
After 1st week
|
After 2nd week
|
After 3rd week
|
After 4th week
|
After 5th week
|
Average Size (cm)
|
Stem
|
Leaves
|
Stem
|
Leaves
|
Stem
|
Leaves
|
Stem
|
Leaves
|
Stem
|
Leaves
|
1
|
Fish bones
|
5.2
|
2
|
10.8
|
4
|
12
|
4.6
|
19
|
5.2
|
21.5
|
5.5
|
2
|
Chicken bones (country)
|
4.4
|
2.5
|
8.2
|
3.7
|
15.2
|
3.8
|
17
|
4
|
17.5
|
4.7
|
3
|
Chicken bones (broiler)
|
3.2
|
1.7
|
4.5
|
1.8
|
8.3
|
3.2
|
14.8
|
3.5
|
15.2
|
4
|
4
|
Goat bones
|
11.5
|
3.9
|
16.2
|
4.35
|
20
|
5.8
|
21.5
|
6.5
|
25
|
7.1
|
5
|
Goat horns
|
11
|
3.1
|
15.9
|
4.34
|
21.5
|
5.6
|
25
|
6
|
20
|
6.9
|
6
|
Cow bones
|
6.5
|
2.4
|
11.9
|
4.85
|
17
|
5.2
|
19.5
|
5.9
|
23.7
|
6.7
|
7
|
Cow horns
|
5.5
|
1.8
|
10.5
|
3.9
|
14.9
|
4.4
|
18.9
|
5
|
21.3
|
5.4
|
8
|
Buffalo bones
|
12
|
3.5
|
18.2
|
5
|
22.4
|
6.1
|
23.7
|
7.4
|
27.2
|
7.6
|
9
|
Buffalo horns
|
11.8
|
3.2
|
18
|
4.98
|
22.2
|
6
|
23.5
|
7.2
|
27
|
7.5
|
10
|
Reference
|
3
|
1.4
|
4
|
1.6
|
8
|
3
|
14.5
|
3.5
|
15
|
3.8
|
In Tables 7–16, the heavy metals adsorption capability of biochar from each sample (bones/horns) was determined. This adsorption capability was checked by the effect of time and concentration on removal of six heavy metals; arsenic (As), cadmium (Cd), chromium (Cr), copper (Cu), nickel (Ni) and lead (Pb). Comparison of the adsorption capability of the biochar made by the different animals, the comparison of bones biochar and horns biochar was done by comparing their adsorption ability by filtration of water solutions containing different heavy metals.
In this study concentration of biochar (made from fish and chicken bones and bones and horns of goat, cow and buffalo) was 1 mg/mL and heavy metal concentration of 100 ppm was used. As time of adsorption was changed from 1 to 6 h, efficiency increases and after 6 h, the efficiency began to decrease. The lowest concentration of heavy metals removal (at 6 h) was recorded by broiler chicken bone biochar while the highest concentration was recorded from the biochar of buffalo bone. In the comparison of biochar made by bones and horns, it was observed that the biochar made by the bones have higher ability of removal of heavy metals than those of horns of same animal. Overall, at 6 h, the lowest removal efficiency of broiler chicken bone biochar was recorded for arsenic (36%) while highest removal efficiency was recorded by the biochar of buffalo bone (75%). Among the metals, the lowest filtration was observed for the arsenic while highest filtration was recorded for lead (for each biochar sample).
Efficiency of biochar was studied at different concentration of six heavy metals (As, Cd, Cr, Cu, Ni and Pb) removed from the artificial wastewater (100 to 500 mg/Kg). All the biochar samples showed highest ability of heavy metals removal at the concentration of 100 mg/Kg while the lowest capability at 500 mg/Kg of artificial wastewater. Among the comparison of all the biochar samples, biochar made from buffalo bone showed the highest filtration for all of the heavy metals from wastewater at 100 mg/Kg (As = 61%, Cd = 71%, Cr = 68%, Cu = 66%, Ni = 63%, Pb = 76%). While the lowest percentage of filtration was recorded by the broiler chicken bone biochar sample (As = 38%, Cd = 46%, Cr = 43%, Cu = 42%, Ni = 39%, Pb = 50%). The removal order of all heavy metals during both time and concentration was same i.e. Pb > Cd > Cr > Cu > Ni > As. While the efficiency order of biochar samples was; buffalo bone biochar > buffalo horn biochar > goat bone biochar > goat horn biochar > cow bone biochar > fish bone biochar > cow horn biochar > country chicken biochar > broiler chicken bone biochar.
Table 7
Removal of heavy metals from wastewater at different time (hour) in fish bone
Time (h)
|
As
Mean ± SD
|
Cd
Mean ± SD
|
Cr
Mean ± SD
|
Cu
Mean ± SD
|
Ni
Mean ± SD
|
Pb
Mean ± SD
|
|
1
|
2 ± 0.29
|
4 ± 0.52
|
3 ± 0.33
|
3 ± 0.34
|
2 ± 0.34
|
5 ± 0.69
|
2
|
10 ± 0.93
|
18 ± 0.88
|
15 ± 0.87
|
13 ± 0.69
|
12 ± 0.68
|
20 ± 1.33
|
3
|
25 ± 1.33
|
35 ± 1.94
|
30 ± 1.41
|
28 ± 1.30
|
26 ± 1.03
|
40 ± 1.63
|
4
|
37 ± 1.51
|
43 ± 1.63
|
41 ± 1.38
|
39 ± 1.63
|
38 ± 1.47
|
49 ± 1.67
|
5
|
42 ± 1.67
|
50 ± 1.76
|
48 ± 1.63
|
45 ± 1.64
|
43 ± 1.30
|
55 ± 1.43
|
6
|
47 ± 1.51
|
57 ± 1.83
|
55 ± 1.30
|
52 ± 1.30
|
50 ± 1.03
|
62 ± 2.09
|
7
|
46 ± 1.17
|
56 ± 1.26
|
54 ± 1.90
|
51 ± 1.63
|
49 ± 1.02
|
61 ± 1.30
|
Table 8
Removal of heavy metals from wastewater by biochar
Conc. mg/Kg
|
As
Mean ± SD
|
Cd
Mean ± SD
|
Cr
Mean ± SD
|
Cu
Mean ± SD
|
Ni
Mean ± SD
|
Pb
Mean ± SD
|
100
|
48 ± 1.60
|
58 ± 2.56
|
56 ± 1.72
|
54 ± 2.06
|
52 ± 1.90
|
64 ± 2.35
|
200
|
43 ± 1.79
|
50 ± 2.60
|
42 ± 1.57
|
45 ± 1.91
|
45 ± 1.60
|
56 ± 1.90
|
300
|
35 ± 1.30
|
38 ± 1.02
|
33 ± 1.14
|
41 ± 1.26
|
39 ± 0.97
|
50 ± 1.72
|
400
|
25 ± 1.46
|
26 ± 0.88
|
25 ± 1.10
|
28 ± 1.02
|
27 ± 0.95
|
42 ± 1.29
|
500
|
15 ± 0.88
|
19 ± 0.79
|
19 ± 0.63
|
18 ± 0.82
|
16 ± 0.68
|
21 ± 0.93
|
Table 9
Removal of heavy metals from wastewater by the biochar of chicken bones at different time (hour)
|
Chicken bone (country)
|
Chicken bone (broiler)
|
Time (h)
|
As
Mean ± SD
|
Cd
Mean ± SD
|
Cr
Mean ± SD
|
Cu
Mean ± SD
|
Ni
Mean ± SD
|
Pb
Mean ± SD
|
As
Mean ± SD
|
Cd
Mean ± SD
|
Cr
Mean ± SD
|
Cu
Mean ± SD
|
Ni
Mean ± SD
|
Pb
Mean ± SD
|
1
|
2 ± 0.36
|
4 ± 0.54
|
3 ± 0.38
|
3 ± 0.36
|
3 ± 0.32
|
4 ± 0.45
|
2 ± 0.25
|
3 ± 0.29
|
3 ± 0.33
|
3 ± 0.35
|
3 ± 0.37
|
4 ± 0.40
|
2
|
10 ± 0.88
|
16 ± 0.79
|
13 ± 0.71
|
12 ± 0.74
|
12 ± 0.89
|
18 ± 0.71
|
8 ± 0.71
|
13 ± 0.88
|
11 ± 0.67
|
10 ± 0.54
|
9 ± 0.55
|
15 ± 0.98
|
3
|
20 ± 1.11
|
31 ± 0.92
|
29 ± 1.03
|
25 ± 1.28
|
24 ± 1.11
|
35 ± 1.33
|
18 ± 0.92
|
25 ± 1.24
|
24 ± 0.84
|
22 ± 0.93
|
19 ± 0.88
|
30 ± 1.33
|
4
|
33 ± 1.36
|
41 ± 1.41
|
38 ± 1.26
|
35 ± 1.33
|
30 ± 1.33
|
45 ± 1.08
|
28 ± 1.37
|
33 ± 1.41
|
30 ± 1.03
|
29 ± 1.03
|
25 ± 1.17
|
39 ± 1.63
|
5
|
40 ± 1.38
|
48 ± 1.33
|
43 ± 1.13
|
40 ± 1.08
|
35 ± 1.08
|
52 ± 1.26
|
35 ± 1.33
|
42 ± 1.60
|
40 ± 1.63
|
37 ± 1.67
|
33 ± 1.21
|
45 ± 1.94
|
6
|
42 ± 1.51
|
52 ± 1.67
|
50 ± 1.25
|
45 ± 1.47
|
43 ± 1.67
|
56 ± 1.76
|
36 ± 1.36
|
44 ± 1.69
|
41 ± 1.74
|
40 ± 1.69
|
37 ± 1.40
|
47 ± 1.67
|
7
|
41 ± 1.21
|
51 ± 1.17
|
49 ± 1.28
|
43 ± 1.41
|
42 ± 1.69
|
54 ± 1.28
|
35 ± 1.41
|
42 ± 1.33
|
40 ± 1.26
|
39 ± 1.17
|
36 ± 1.03
|
45 ± 1.72
|
Table 10
Removal of heavy metals from wastewater by the biochar of chicken bones
|
Chicken bone (country)
|
Chicken bone (broiler)
|
Conc. mg/Kg
|
As
Mean ± SD
|
Cd
Mean ± SD
|
Cr
Mean ± SD
|
Cu
Mean ± SD
|
Ni
Mean ± SD
|
Pb
Mean ± SD
|
As
Mean ± SD
|
Cd
Mean ± SD
|
Cr
Mean ± SD
|
Cu
Mean ± SD
|
Ni
Mean ± SD
|
Pb
Mean ± SD
|
|
100
|
46 ± 1.41
|
54 ± 1.72
|
52 ± 1.74
|
47 ± 1.46
|
45 ± 1.50
|
57 ± 1.99
|
38 ± 1.30
|
46 ± 1.50
|
43 ± 1.47
|
42 ± 1.08
|
39 ± 1.20
|
50 ± 1.78
|
|
200
|
41 ± 1.33
|
49 ± 1.63
|
45 ± 1.33
|
41 ± 1.07
|
36 ± 0.89
|
53 ± 1.76
|
35 ± 1.08
|
43 ± 1.11
|
40 ± 1.03
|
39 ± 1.11
|
35 ± 1.01
|
46 ± 1.36
|
|
300
|
35 ± 1.03
|
42 ± 1.21
|
39 ± 1.44
|
37 ± 1.07
|
30 ± 1.03
|
45 ± 1.41
|
28 ± 0.89
|
34 ± 1.03
|
32 ± 0.93
|
31 ± 0.94
|
27 ± 0.86
|
40 ± 1.08
|
|
400
|
22 ± 0.86
|
33 ± 1.11
|
30 ± 0.97
|
27 ± 0.98
|
25 ± 0.82
|
36 ± 1.03
|
18 ± 0.74
|
27 ± 0.88
|
25 ± 0.92
|
24 ± 0.86
|
20 ± 0.92
|
32 ± 1.03
|
|
500
|
12 ± 0.69
|
18 ± 0.97
|
14 ± 0.74
|
14 ± 0.72
|
13 ± 0.83
|
19 ± 0.85
|
10 ± 0.46
|
14 ± 0.63
|
12 ± 0.61
|
12 ± 0.60
|
11 ± 0.69
|
17 ± 0.74
|
|
Table 11
Removal of heavy metals from wastewater by the biochar of goat bones and horns at different time (hour)
|
Goat bone
|
Goat horns
|
Time (h)
|
As
Mean ± SD
|
Cd
Mean ± SD
|
Cr
Mean ± SD
|
Cu
Mean ± SD
|
Ni
Mean ± SD
|
Pb
Mean ± SD
|
As
Mean ± SD
|
Cd
Mean ± SD
|
Cr
Mean ± SD
|
Cu
Mean ± SD
|
Ni
Mean ± SD
|
Pb
Mean ± SD
|
1
|
4 ± 0.47
|
5 ± 0.46
|
5 ± 0.35
|
5 ± 0.35
|
5 ± 0.34
|
6 ± 0.34
|
4 ± 0.59
|
5 ± 0.88
|
5 ± 0.52
|
5 ± 0.69
|
5 ± 0.63
|
6 ± 0.71
|
2
|
15 ± 0.66
|
20 ± 1.33
|
18 ± 0.84
|
18 ± 0.66
|
16 ± 0.66
|
22 ± 0.98
|
11 ± 0.88
|
16 ± 1.03
|
14 ± 0.84
|
14 ± 0.92
|
14 ± 0.98
|
20 ± 0.98
|
3
|
28 ± 0.71
|
39 ± 0.93
|
35 ± 1.56
|
34 ± 0.92
|
30 ± 1.37
|
40 ± 1.37
|
24 ± 1.11
|
35 ± 1.37
|
31 ± 1.11
|
30 ± 1.37
|
29 ± 1.17
|
38 ± 0.82
|
4
|
45 ± 0.94
|
52 ± 1.28
|
50 ± 2.04
|
48 ± 1.26
|
48 ± 1.60
|
54 ± 1.76
|
40 ± 1.60
|
48 ± 1.63
|
43 ± 1.99
|
42 ± 1.67
|
40 ± 1.43
|
50 ± 1.36
|
5
|
53 ± 1.23
|
61 ± 1.03
|
58 ± 1.43
|
56 ± 1.67
|
56 ± 1.28
|
65 ± 2.16
|
48 ± 1.78
|
59 ± 1.79
|
54 ± 2.04
|
53 ± 1.90
|
49 ± 1.90
|
61 ± 1.97
|
6
|
58 ± 1.42
|
69 ± 2.04
|
65 ± 2.16
|
63 ± 1.76
|
60 ± 2.09
|
73 ± 2.23
|
53 ± 2.23
|
63 ± 2.06
|
60 ± 2.50
|
58 ± 2.07
|
57 ± 1.60
|
68 ± 2.51
|
7
|
57 ± 1.28
|
68 ± 1.33
|
64 ± 1.41
|
62 ± 1.60
|
59 ± 1.94
|
72 ± 1.86
|
52 ± 1.63
|
62 ± 2.00
|
59 ± 2.19
|
57 ± 1.90
|
56 ± 1.76
|
67 ± 2.75
|
Table 12
Removal of heavy metals from wastewater by biochar of goat bones and horns at concentration of heavy metals
|
Goat bone
|
Goat horns
|
Conc. mg/Kg
|
As
Mean ± SD
|
Cd Mean ± SD
|
Cr Mean ± SD
|
Cu Mean ± SD
|
Ni Mean ± SD
|
Pb Mean ± SD
|
As Mean ± SD
|
Cd Mean ± SD
|
Cr Mean ± SD
|
Cu
Mean ± SD
|
Ni Mean ± SD
|
Pb Mean ± SD
|
|
100
|
59 ± 1.90
|
70 ± 2.95
|
67 ± 1.63
|
65 ± 1.51
|
61 ± 2.04
|
75 ± 2.14
|
55 ± 1.63
|
65 ± 2.53
|
62 ± 2.28
|
60 ± 1.84
|
58 ± 1.96
|
69 ± 2.25
|
|
200
|
54 ± 1.60
|
63 ± 1.80
|
60 ± 1.60
|
58 ± 1.69
|
57 ± 1.90
|
67 ± 1.86
|
49 ± 1.33
|
60 ± 1.72
|
56 ± 1.90
|
55 ± 1.41
|
50 ± 1.41
|
63 ± 1.78
|
|
300
|
46 ± 1.76
|
54 ± 1.67
|
52 ± 1.87
|
50 ± 1.33
|
50 ± 1.41
|
55 ± 1.90
|
42 ± 1.41
|
50 ± 1.30
|
45 ± 1.60
|
44 ± 1.33
|
42 ± 1.28
|
52 ± 1.74
|
|
400
|
30 ± 1.03
|
41 ± 1.26
|
37 ± 0.99
|
36 ± 1.02
|
32 ± 1.14
|
42 ± 1.63
|
26 ± 0.93
|
37 ± 1.08
|
33 ± 0.98
|
32 ± 0.97
|
31 ± 1.03
|
40 ± 1.10
|
|
500
|
17 ± 0.89
|
21 ± 0.88
|
20 ± 0.71
|
20 ± 0.98
|
17 ± 0.88
|
24 ± 1.02
|
13 ± 0.56
|
17 ± 0.66
|
16 ± 0.75
|
16 ± 0.56
|
16 ± 0.83
|
22 ± 1.02
|
|
Table 13
Removal of heavy metals from wastewater by the biochar of cow bones and horns at different time (hour)
|
Cow bones
|
Cow horns
|
Time (h)
|
As
Mean ± SD
|
Cd
Mean ± SD
|
Cr
Mean ± SD
|
Cu
Mean ± SD
|
Ni
Mean ± SD
|
Pb
Mean ± SD
|
As
Mean ± SD
|
Cd
Mean ± SD
|
Cr
Mean ± SD
|
Cu
Mean ± SD
|
Ni
Mean ± SD
|
Pb
Mean ± SD
|
1
|
2 ± 0.37
|
5 ± 0.66
|
4 ± 0.63
|
4 ± 0.52
|
4 ± 0.38
|
6 ± 0.82
|
2 ± 0.28
|
4 ± 0.71
|
4 ± 0.46
|
4 ± 0.38
|
4 ± 0.36
|
5 ± 0.48
|
2
|
14 ± 0.71
|
18 ± 0.88
|
16 ± 0.99
|
16 ± 0.80
|
15 ± 0.71
|
20 ± 0.88
|
12 ± 0.52
|
17 ± 0.80
|
16 ± 0.88
|
16 ± 1.02
|
15 ± 0.66
|
0.82 ± 0.81
|
3
|
28 ± 1.03
|
32 ± 1.37
|
31 ± 1.11
|
30.5 ± 1.13
|
29 ± 1.03
|
36 ± 1.03
|
25 ± 1.21
|
33 ± 1.02
|
32 ± 1.03
|
31 ± 1.11
|
29 ± 1.02
|
35 ± 1.03
|
4
|
1.02
|
45 ± 1.60
|
44 ± 1.33
|
42 ± 1.17
|
41 ± 1.33
|
50 ± 1.37
|
34 ± 1.30
|
45 ± 1.36
|
44 ± 1.38
|
43 ± 1.03
|
41 ± 1.21
|
48 ± 1.22
|
5
|
45 ± 1.37
|
54 ± 1.69
|
52 ± 1.39
|
51 ± 1.43
|
49 ± 1.17
|
56 ± 1.26
|
42 ± 1.41
|
52 ± 1.33
|
51 ± 1.67
|
49 ± 1.11
|
47 ± 1.38
|
56 ± 1.60
|
6
|
49 ± 1.68
|
60 ± 2.11
|
58 ± 1.60
|
55 ± 1.10
|
52 ± 1.60
|
65 ± 2.23
|
47 ± 2.07
|
58 ± 1.86
|
56 ± 2.07
|
53 ± 1.33
|
50 ± 1.33
|
63 ± 1.86
|
7
|
48 ± 1.50
|
59 ± 1.75
|
56 ± 1.26
|
54 ± 1.33
|
51 ± 1.05
|
64 ± 1.36
|
46 ± 1.84
|
57 ± 1.72
|
55 ± 1.47
|
52 ± 1.37
|
49 ± 1.90
|
62 ± 1.72
|
Table 14
Removal of heavy metals from wastewater by biochar of cow bones and horns at concentration of heavy metals
|
Cow bones
|
Cow horns
|
Conc. mg/Kg
|
As
Mean ± SD
|
Cd
Mean ± SD
|
Cr
Mean ± SD
|
Cu
Mean ± SD
|
Ni
Mean ± SD
|
Pb
Mean ± SD
|
As
Mean ± SD
|
Cd
Mean ± SD
|
Cr
Mean ± SD
|
Cu
Mean ± SD
|
Ni
Mean ± SD
|
Pb
Mean ± SD
|
100
|
51 ± 1.30
|
62 ± 1.91
|
59 ± 2.23
|
57 ± 1.75
|
53 ± 1.86
|
67 ± 2.28
|
49 ± 1.87
|
60 ± 3.16
|
57 ± 2.46
|
55 ± 2.28
|
51 ± 2.61
|
65 ± 2.83
|
200
|
46 ± 1.24
|
53 ± 1.60
|
50 ± 1.86
|
50 ± 1.33
|
47 ± 1.86
|
58 ± 2.64
|
41 ± 1.76
|
54 ± 2.66
|
52 ± 2.14
|
50 ± 1.74
|
45 ± 2.38
|
57 ± 2.73
|
300
|
37 ± 1.16
|
45 ± 1.33
|
42 ± 1.28
|
41 ± 1.37
|
37 ± 1.41
|
51 ± 1.75
|
35 ± 1.36
|
46 ± 1.96
|
44 ± 1.76
|
42 ± 1.80
|
40 ± 1.36
|
49 ± 2.32
|
400
|
27 ± 0.97
|
30 ± 1.11
|
30 ± 1.11
|
29 ± 1.28
|
27 ± 1.33
|
38 ± 1.41
|
26 ± 1.28
|
35 ± 1.67
|
34 ± 1.36
|
31 ± 1.25
|
30 ± 1.03
|
36 ± 1.44
|
500
|
16 ± 0.72
|
19 ± 0.86
|
17 ± 0.60
|
16 ± 0.95
|
16 ± 0.98
|
22 ± 0.98
|
14 ± 0.76
|
17 ± 0.92
|
15 ± 0.97
|
15 ± 0.83
|
15 ± 0.91
|
20 ± 1.033
|
Table 15
Removal of heavy metals from wastewaterby biochar of buffalo bones and horns at different time (hour)
|
Buffalo bones
|
Buffalo horns
|
Time (h)
|
As
Mean ± SD
|
Cd
Mean ± SD
|
Cr
Mean ± SD
|
Cu
Mean ± SD
|
Ni
Mean ± SD
|
Pb
Mean ± SD
|
As
Mean ± SD
|
Cd
Mean ± SD
|
Cr
Mean ± SD
|
Cu
Mean ± SD
|
Ni
Mean ± SD
|
Pb
Mean ± SD
|
|
1
|
4 ± 0.46
|
6 ± 0.55
|
5 ± 0.06
|
5 ± 0.46
|
4 ± 0.37
|
7 ± 0.47
|
4 ± 0.46
|
5 ± 0.41
|
5 ± 0.41
|
5 ± 0.46
|
4 ± 0.47
|
6 ± 0.55
|
|
2
|
16 ± 0.93
|
22 ± 0.66
|
20 ± 0.99
|
18 ± 0.98
|
17 ± 0.71
|
23 ± 0.99
|
15 ± 0.92
|
18 ± 0.88
|
17 ± 0.71
|
17 ± 0.94
|
16 ± 0.71
|
20 ± 1.08
|
|
3
|
32 ± 1.02
|
40 ± 1.26
|
38 ± 1.26
|
36 ± 1.03
|
35 ± 1.02
|
42 ± 1.33
|
30 ± 1.17
|
37 ± 1.33
|
36 ± 0.88
|
34 ± 0.95
|
33 ± 0.99
|
39 ± 1.40
|
|
4
|
44 ± 1.41
|
54 ± 1.33
|
52 ± 1.38
|
50 ± 1.33
|
48 ± 1.63
|
56 ± 1.38
|
42 ± 1.33
|
51 ± 1.30
|
49 ± 1.60
|
47 ± 1.33
|
45 ± 1.30
|
52 ± 1.14
|
|
5
|
55 ± 1.97
|
65 ± 1.33
|
63 ± 1.72
|
61 ± 1.94
|
59 ± 1.94
|
68 ± 1.75
|
52 ± 1.40
|
62 ± 1.76
|
60 ± 1.67
|
57 ± 1.28
|
54 ± 1.60
|
66 ± 1.90
|
|
6
|
60 ± 2.23
|
70 ± 1.63
|
67 ± 1.86
|
65 ± 2.42
|
62 ± 1.86
|
75 ± 2.32
|
56 ± 1.75
|
66 ± 1.80
|
64 ± 1.63
|
62 ± 1.41
|
59 ± 2.04
|
71 ± 1.63
|
|
7
|
59 ± 2.11
|
69 ± 2.06
|
66 ± 2.25
|
64 ± 1.63
|
61 ± 2.23
|
74 ± 1.90
|
55 ± 1.36
|
65 ± 1.43
|
63 ± 1.72
|
61 ± 1.41
|
60 ± 1.70
|
70 ± 1.96
|
|
Table 16
Removal of heavy metals by biochar of buffalo bones and horns at concentration of HM
|
Buffalo bones
|
Buffalo horns
|
Conc. mg/Kg
|
As
Mean ± SD
|
Cd
Mean ± SD
|
Cr
Mean ± SD
|
Cu
Mean ± SD
|
Ni
Mean ± SD
|
Pb
Mean ± SD
|
As
Mean ± SD
|
Cd
Mean ± SD
|
Cr
Mean ± SD
|
Cu
Mean ± SD
|
Ni
Mean ± SD
|
Pb
Mean ± SD
|
|
100
|
61 ± 2.94
|
71 ± 3.72
|
68 ± 2.14
|
66 ± 2.64
|
63 ± 2.50
|
76 ± 3.31
|
57 ± 2.73
|
67 ± 3.33
|
65 ± 3.09
|
63 ± 2.29
|
60 ± 3.46
|
72 ± 3.37
|
|
200
|
56 ± 1.75
|
64 ± 2.64
|
63 ± 2.07
|
62 ± 2.48
|
60 ± 2.71
|
70 ± 2.61
|
53 ± 2.07
|
63 ± 3.08
|
61 ± 3.31
|
55 ± 2.43
|
53 ± 2.34
|
65 ± 2.42
|
|
300
|
45 ± 1.86
|
55 ± 1.86
|
53 ± 2.16
|
51 ± 2.17
|
50 ± 2.59
|
57 ± 2.53
|
42 ± 1.75
|
52 ± 2.48
|
50 ± 2.88
|
45 ± 1.67
|
44 ± 2.14
|
53 ± 2.06
|
|
400
|
33 ± 1.41
|
42 ± 1.33
|
40 ± 1.75
|
38 ± 1.75
|
36 ± 1.43
|
43 ± 1.69
|
31 ± 1.41
|
38 ± 1.41
|
37 ± 1.41
|
33 ± 1.41
|
32 ± 1.60
|
40 ± 2.42
|
|
500
|
18 ± 0.94
|
24 ± 1.11
|
21 ± 1.03
|
19 ± 1.03
|
18 ± 0.86
|
25 ± 1.33
|
18 ± 1.03
|
20 ± 1.02
|
19 ± 0.88
|
19 ± 1.08
|
18 ± 0.88
|
23 ± 1.02
|
|
According to results, fish bones, chicken bones (country), chicken bones (broiler) and goat trotters showed antibacterial activities against Enterococcus faecalis while fish bones were found to be active against Staphylococcus aureus and Bacillus subtilis. Chicken bones (country) and goat trotters showed antibacterial activities against Salmonella typhi while goat trotters were found to be active against Pseudomonas aeruginosa. However, none of the bones had any bioactivity against E. coli. Fish bones showed antifungal activities against Candida albicans while none of the bones were found to be active against plant pathogenic fungi (Table 17–20; Fig. 4–6).
Table 17
Antibacterial activity of different calcinated bones against Gram positive bacteria.
Sample names
|
Zone of inhibition (mm)
|
Staphylococcus aureus
|
Bacillus subtilis
|
Enterococcus faecalis
|
Fish bones
|
19 mm
|
20 mm
|
10 mm
|
Chicken bones (country)
|
-ve
|
-ve
|
20 mm
|
Chicken bones (broiler)
|
-ve
|
-ve
|
15 mm
|
Goat trotters
|
-ve
|
-ve
|
20 mm
|
Goat horns
|
-ve
|
-ve
|
-ve
|
Cow bones
|
-ve
|
-ve
|
-ve
|
Buffaloes horns
|
-ve
|
-ve
|
-ve
|
-ve = no zone of growth inhibition.
|
Table 18
Antibacterial activity of different calcinated bones against Gram negative bacteria
Sample names
|
Zone of inhibition (mm)
|
Escherichia coli
|
Pseudomonas aeruginosa
|
Salmonella typhi
|
Fish bones
|
-ve
|
-ve
|
-ve
|
Chicken bones (country)
|
-ve
|
-ve
|
10mm
|
Chicken bones (broiler)
|
-ve
|
-ve
|
-ve
|
Goat trotters
|
-ve
|
15 mm
|
12 mm
|
Goat horns
|
-ve
|
-ve
|
-ve
|
Cow bones
|
-ve
|
-ve
|
-ve
|
Buffaloes horns
|
-ve
|
-ve
|
-ve
|
-ve = no zone of growth inhibition.
|
Table 19
Antifungal activity of different calcinated bones against fungi
Name of samples
|
Zone of inhibition (mm)
|
Candida albicans
|
Candida glabrata
|
Mucor sp.
|
Aspergillus niger
|
Aspergillus flavus
|
Fish bones
|
10 mm
|
-ve
|
-ve
|
-ve
|
-ve
|
Chicken bones (country)
|
-ve
|
-ve
|
-ve
|
-ve
|
-ve
|
Chicken bones (broiler)
|
-ve
|
-ve
|
-ve
|
-ve
|
-ve
|
Goat trotters
|
-ve
|
-ve
|
-ve
|
-ve
|
-ve
|
Goat horns
|
-ve
|
-ve
|
-ve
|
-ve
|
-ve
|
Cow bones
|
-ve
|
-ve
|
-ve
|
-ve
|
-ve
|
Buffaloes horns
|
-ve
|
-ve
|
-ve
|
-ve
|
-ve
|
Table 20
Antifungal activity of different calcinated bones against plant pathogenic fungi
Sample names
|
Zone of inhibition (mm)
|
Alternaria sp.
|
Drechslera sp.
|
Humicola sp.
|
Fish bones
|
-ve
|
-ve
|
-ve
|
Hen bones (country)
|
-ve
|
-ve
|
-ve
|
Hen bones (broiler)
|
-ve
|
-ve
|
-ve
|
Goat trotters
|
-ve
|
-ve
|
-ve
|
Goat horns
|
-ve
|
-ve
|
-ve
|
Cow bones
|
-ve
|
-ve
|
-ve
|
Buffaloes horns
|
-ve
|
-ve
|
-ve
|