Phyton-International Journal of Experimental Botany |
DOI: 10.32604/phyton.2021.011413
ARTICLE
Poultry Manure as an Organic Fertilizer with or without Biochar Amendment: Influence on Growth and Heavy Metal Accumulation in Lettuce and Spinach and Soil Nutrients
1Department of Botany, University of Balochistan, Quetta, 87300, Pakistan
2Department of Natural Resource Sciences, McGill University, Quebec, H9X 3V9, Canada
3Department of Chemistry, University of Balochistan, Quetta, 87300, Pakistan
4Department of Zoology, University of Balochistan, Quetta, 87300, Pakistan
*Corresponding Author: Shamim Gul. Email: shamim.gul@mail.mcgill.ca
Received: 07 May 2020; Accepted: 06 July 2020
Abstract: This pot-based study investigated the influence of poultry manure and 1:1 mixture of poultry manure + biochar (produced from farmyard manure [FYM] or wood), on the biomass production and concentration of heavy metals in leaves of lettuce and spinach. The concentration of mineral nitrogen (N) and soluble inorganic phosphorus (P) of soils cultivated with these vegetables was also investigated. The application of poultry manure or FYM biochar in soil as 10% (equivalent to 60 t ha–1, an estimated 1726.8 kg ha–1 N in poultry manure and 1353.9 kg ha–1 N in FYM) and 15% amendment (equivalent to 90 t ha–1, an estimated 2590.2 kg ha–1 N in poultry manure and 2030.8 kg ha–1 N in FYM) significantly decreased biomass production of lettuce as compared to control (no fertilizer added) treatment. However, mixture of poultry manure with wood-derived biochar at both application rates (i.e., 10% and 15%) and with FYM biochar at lower application rate (i.e., 10%) caused 2–3-fold increase in aboveground plant biomass and 2–14-fold increase in root biomass (p < 0.05). Furthermore, as compared to control treatment, a significant ~2–3-fold increase in aboveground plant biomass was also observed in response to mixture of poultry manure with wood-derived and FYM derived biochars at 10% amendment rates. As compared to control treatment, concentration of mineral N and soluble inorganic P were higher in soils of all other treatments. In spinach, amendment of poultry manure or its co-amendment with biochar of FYM significantly increased aboveground plant biomass at 7% (equivalent to 42 t ha–1) as compared to 3% and 5% amendment rates (equivalent to 18 and 30 t ha–1 respectively). The concentration of soil mineral N and soil soluble mineral P was not different between treatments. In lettuce, wood-derived biochar did not reduce concentration of heavy metals (i.e., manganese (Mn), copper (Cu), iron, (Fe), cadmium (Cd), lead (Pb), nickel (Ni) and cobalt (Co) than FYM-derived biochar while in spinach, as compared to poultry manure, co-amendment of poultry manure with wood-derived biochar reduced concentration of heavy metals, indicating differential responses of crops to organic amendments.
Keywords: Biochar; poultry manure; lettuce; spinach
Poultry industry started in Pakistan in 1962, this industry turned into an important sector of livestock in Pakistan, providing employment to 1.5 million people and contributed in 1.4% gross domestic production (GDP) of the country in 2017–2018 [1]. To date, twenty-five thousand poultry farms are reported in Pakistan [2]. The production of cow meat is also increasing in Pakistan [1]. The growing industry of poultry and livestock production in Pakistan is also generating production of manure with the same rate. The utilization of manure produced from poultry and livestock as organic fertilizer in soil can be a means of its proper disposal and its prevention to be wasted, besides of its high potential to be utilized in biogas production for energy generation [2]. Poultry manure as a fertilizer was a part of organic crop production from centuries and it has been considered as the most suitable among the natural fertilizers because it has high nitrogen content in it [3].
Biochar is pyrogenous biomass that is made from burning biomass (e.g., stover, manure, bark, nutshells, algae, animal bones, etc.) in oxygen-free or oxygen-deficient conditions [4]. Biochar is hygroscopic can retain and attract water, because of its high surface area and porous structure [5]. It prevents the leaching of nutrients from the soil [6]. When biochar is mixed with compost or manure, it absorbs nutrients and become a slow-release fertilizer [7]. Many beneficial effects of biochar depend upon its different properties [8]. The utility of these manures can be maximized with the help of their co-amendment with biochar, which is reported to be more beneficial for soil quality and crop production than when these organic substances (manures and biochar) are amended alone [7,9–11]. Biochar-manure mixture can be more economical when it is applied in the root zone of crops as it will be used in many times less amount than when it is applied thoroughly in soil [12].
Lettuce (Lactuca sativa) and spinach (Spinacia oleracea) are two important vegetable crops of Pakistan [13]. There is wealth of reports that demonstrate positive influence of manures and biochar-manure mixture on yield of these two important crops [14–17]. However, because in developing countries, there is no proper management of utilization of manures, they can contribute significantly in environmental pollution especially emission of greenhouse gases [18,19]. For this reason, there is need to assess if high application rates of these manures in soil with combination of biochar can have a positive influence on the yield of vegetable crops. Biochar due to its highly porous nature, captures nutrients and act as slow-release fertilizer [10] and therefore, if mixed with manures can attenuate high application rate-related negative influence of manure fertilizers. Such a study will help farmers to utilize these two important biowastes, which will in return help prevent environmental hazards related to their waste in environment. Objectives of the present study were to evaluate the influence of poultry manure and farmyard manure (FYM) as organic fertilizers, amended in soil alone or as a mixture with biochar, thoroughly mixed in soil or applied in the root zone of lettuce, on growth, water use efficiency (WUE) and concentration of heavy metals, i.e., manganese (Mn), copper (Cu), iron, (Fe), cadmium (Cd), lead (Pb), nickel (Ni) and cobalt (Co) in lettuce. Two types of biochars were used; biochar produced from wood and biochar produced from cow manure. We also tested poultry manure as organic amendment in soil alone or mixed with wood-derived biochar or biochar produced from FYM at lower rates than the rates applied to lettuce (as in lettuce high application rates of manure amendment reduced growth) on growth, WUE and concentration of heavy metals (as mentioned above) in spinach. Soil quality parameters such as organic matter, pH, electrical conductivity and concentration of mineral nitrogen (N) and soluble mineral phosphorus (P) of soils after harvest of crops were also tested. Our main hypotheses are mixture of poultry manure with biochar increases 1) plant growth performance and 2) reduces concentration of heavy metals in soil. Dikinya et al. [20] observed higher yield of spinach in Luvic Calcisol at 10% amendment rate of poultry manure as compared to 5% amendment rate. Yang et al. [21] used 20% amendment rate of poultry manure in their research that was related to evaluate abundance of endopytic bacteria in Pakchoi and antibiotic resistant bacteria in soil planted with Pakchoi. Hao et al. [22] observed approximately 15 times greater fresh biomass production of Pakchoi in response to amendment of poultry manure at 8% amendment rate. Therefore, we used 10% and 15% amendment rates for poultry manure for spinach. The main reason of using these high application rates was to provide an insight to local farmers to maximize the use of this manure for agriculture purpose, which can in return help substantially in reducing environmental pollution caused by its waste.
2.1 Biochar Production and Experimental Setup
In this study slow pyrolyzed wood-based biochar was purchased from the local timber market of Quetta, Balochistan, Pakistan. This biochar was produced in kilns, where the temperature is between 300°C and 500°C [23,24]. The biochar produced from cow manure (FYM in text) was prepared by the burning of dry cow manure under the oxygen-deficient conditions in kilns. The poultry manure in this project was taken from poultry farm in Quetta, Pakistan. Fresh poultry manure was kept under sunlight to properly dry it. The biochar-manure mixture was prepared as 1:1 w/w biochar:manure ratio on dry bases. Properties of biochars and poultry manure are provided in Tab. 1.
Sandy loam soil was selected for this study. The soil was air-dried completely and sieved through 2 mm mesh, 600 g soil was used for each plastic pot of lettuce plant. Commercially available plastic pots of 08 cm diameter and 15 cm height were filled with soil. For lettuce crop, experiment was designed as factorial with three factors; 1) fertilizer type 2) amendment rate and 3) fertilizer placement (thoroughly mixed or placed as root-zone fertilizer). For lettuce, five treatments except control treatment were selected for thoroughly mix type as 1) control (no organic amendment), 2) wood-based biochar, 3) poultry manure, 4) mixture of wood-based biochar-poultry manure as 1:1 ratio and 5) mixture of FYM biochar-poultry manure as 1:1 ratio. Each organic fertilizer was applied at two amendment rates10% (equivalent to 60 t ha−1) and 15% (equivalent to 90 t ha−1). Based on published reports regarding concentration of total nitrogen in poultry and FYM presented in Supporting Information Tab. S1, these amendment rates may contain respectively an estimated 1726.8 kg ha−1 N and 2590.2 kg ha−1 N input for poultry manure and respectively an estimated 1353.9 kg ha−1 N and 2030.85 kg ha−1 N input for FYM [25–29]. Previous reports suggest that the yield of lettuce was higher at higher application rate of synthetic fertilizer as 200 kg ha−1 [30] and 271 kg ha−1 [31]. Three treatments were selected for root zone fertilizer trial; 1) poultry manure, 2) mixture of wood-based biochar-chicken manure (mixed at 1:1 ratio) and 3) mixture of FYM biochar-chicken manure (mixed at 1:1 ratio). Each fertilizer was applied at three amendment rates, i.e., 5% (equivalent to 30 t ha−1), 10%, and 15%). The pots were filled with soil, then an amount of fertilizer equivalent to 5%, 10% or 15% rate were surface applied and the fertilizers were then covered with soil. Each treatment had three replications.
For spinach, only fertilizer as thoroughly-mixed treatments were used and the amendment rates were kept lower than the ones for lettuce. The experimental design was factorial with factors; 1) fertilizer type and 2) amendment rate. This was because for lettuce, root-zone fertilizer and manures at higher rates (10% and 15%) without biochar mixture had a negative influence on crop growth. For lettuce following treatments were made; 1) poultry manure, 2) mixture of FYM biochar-poultry manure (mixed at 1:1 ratio) and the mixture of wood-based biochar—poultry manure (mixed at 1:1 ratio). Each fertilizer was amended at 3% (~18 t ha−1), 5% (~30 t ha−1) and 7% (~42 t ha−1) amendment rate in soil. These amendment rates are approximately equal to 290 kg ha−1, 483 kg ha−1 and 676 kg ha−1 of total N input respectively from poultry manure and 406 kg ha−1, 677 kg ha−1 and 948 kg ha−1 of total N input from FYM manure according to the Supporting Information Tab. S1. Previous studies showed a positive linear growth rate of spinach in response to the amendment rate of synthetic fertilizer and the maximum yield was reported at highest application rate of 200 kg ha−1 [32,33]. All treatments had three replications. Treatments and abbreviations of each treatment are given in Tab. 2.
2.2 Cultivation and Harvesting
Lettuce seeds were purchased from the local market of Quetta city and broadcasted in the third week of November over all the pots. After two weeks of seed germination, the seedlings were thinned to six seedlings per pot [14]. The water use efficiency was carried out after two weeks of germination, till the time of harvesting the pots were irrigated with an equal amount of water at alternate days and once in a week were adjusted to ~60% water filled pore space (WFPS) as described in Gul et al. [34].
Spinach seeds for this research were purchased from the local market of Quetta city and were broadcasted in last week of November over all the pots [15]. After two weeks of seed germination, the seedlings were thinned to 10 seedlings per pot. The water use efficiency was carried out after two weeks of germination, till the time of harvesting the pots were irrigated with an equal amount of water at alternate days and once in a week were adjusted to approximately 60% WFPS as described in Gul et al. [34]. Water use efficiency was calculated by using the following formula [35]:
Water Use Efficiency = Plant biomass/Amount of water (L) provided.
2.3 Assessment of Growth Performance of Plants
At the time of harvesting (~7 weeks after germination), the above-ground biomass of plant and its roots were separated by cutting each pot from two sides with the help of a sharp scissor. The soil root system present in that was removed very carefully without damaging the root system. Roots were carefully washed. The above-ground biomass and roots were air-dried for 48 hours, afterword kept in an oven at 60°C for 48 hours then dry biomass and root weight was measured [15,36].
2.4 Assessment of Heavy Metals
Heavy metals assessment was carried out by burning 1 g sample of above-ground biomass of both lettuce and spinach plant tissues, wood biochar, cow manure biochar and chicken manure at 500°C in furnace till ash formation. Ash was then transferred in bottles, containing 30 ml distilled water and 1.5 ml of concentrated hydrochloric acid as described in Ghori et al. [15]. Heavy metals concentration of plant tissues, cow manure biochar, wood biochar, and chicken manure samples were analyzed by using flame atomic absorption spectrophotometer (AA 7000 Shimadzu). Protocol and instrumental conditions for heavy metal analysis are carried out following protocol of Khan et al. [37].
2.5 Assessment of pH, E.C, mineral Nitrogen and soluble Phosphorus in soil
The assessment of Nitrogen and soluble Phosphorus was carried out by extracting the soil samples with 2M KCl solution as it was described in Gul et al. [34]. Total mineral nitrogen as ammonium (NH4+) and nitrate (NO3−) was assessed by following the protocol of Sims et al. [42]. Soluble inorganic Phosphorus was analyzed by following the process as described in D’Angelo et al. [43]. The extracts of soil samples were analyzed on UV-visible spectrophotometer (Shimadzu UV-700) for nitrogen and soluble phosphorus. The pH and electrical conductivity of soil were analyzed by mixing soil samples in distilled water by 1:2 w/v ratio as described in Dupuis et al. [44].
All the data sets were assessed for normal distribution followed by one-way analysis of variance (ANOVA) least significance difference (LSD) test. Statistical evaluation of all results was carried out by using the most advanced computerized statistical CoStat software (version 6.400) and Microsoft Excel.
3.1 Plant Growth Performance and Water Use Efficiency
In lettuce plant growth performance and water use efficiency (WUE) was higher in organic amendments that were thoroughly mixed as compared to organic amendments in the root zone of plants. Organic amendments tend to increase the shoot biomass, root biomass and water use efficiency (WUE) of lettuce (p < 0.05). Amendment of poultry manure and FYM biochar significantly reduced biomass production than control; however, when mixed with wood-based biochar at both application rates and FYM biochar at lower rates caused 2–3-fold increase in aboveground plant biomass and 2–14-fold increase in root biomass (Figs. 1 and 2; p < 0.05; raw data in Supporting Information Tab. S2). Furthermore, mixture of poultry manure with wood-derived biochar and FYM-derived biochar at 10% amendment rate significantly increased aboveground plant biomass (p < 0.05; Fig. 2). Application of poultry manure and FYM biochar at the root zone of plants significantly reduced growth pf plants as compared to the treatments where these fertilizers were thoroughly mixed in soil, while amendment of wood-based biochar did not ameliorate their negative effect (Figs. 1 and 2; p < 0.05). The WUE of plants were significantly higher as compared to control in response to the co-amendment of wood-based and FYM biochar with poultry manure (Fig. 2; p < 0.05; raw data in Supporting Information Tab. S3). For spinach, aboveground plant biomass was significantly lower in response to the amendment of poultry manure at lower application rate (3%) than other treatments (Figs. 1 and 2; p < 0.05; raw data in Tab. S4). Root biomass did not show the same trend as for aboveground plant biomass (Fig. 2; p < 0.05).The WUE of spinach showed positive relation to the aboveground plant biomass (Fig. 2; p < 0.05; raw data in Tab. S5).
In lettuce-grown soil, as compared to control, concentration of mineral N and soluble mineral P was significantly higher in response to organic amendments (Fig. 3; p < 0.05; raw data in Supporting Information Tabs. S6 and S7).
In spinach-grown soil, concentration of mineral N was significantly higher in response to the amendment of poultry manure and mixture of FYM + poultry manure at 7% rate as compared to the treatments of 3% amendment rates for poultry manure and mixture of FYM + poultry manure (Fig. 3; p < 0.05; raw data in Supporting Information Tab. S8). The concentration of P was significantly higher in response to the amendment of poultry manure at 7% rate and mixture of wood-based biochar and poultry manure at 5% amendment rate than the soil amended with mixture of wood-based biochar with poultry manure at 5% and 7% rates (Fig. 3; p < 0.05; raw data in Supporting Information Tab. S9).
3.3 Concentration of Heavy Metals in Aboveground Plant Tissues
The concentration of heavy metals (manganese [Mn], copper [Cu], iron [Fe], cadmium [Cd], lead [Pb], nickle [Ni] and cobalt [Co] was higher in tissues of lettuce plants grown in soils amended with organic fertilizers as compared to control (Tab. 3). However, the nutrient use efficiency (NUE) of plants for heavy metals tend to be higher in response to organic amendments as compared to control (Tab. 3). In spinach, increasing the amendment rate of poultry manure increased the concentration of heavy metals; however, amendment of mixture of wood-based biochar with poultry manure reduced concentration of heavy metals (Tab. 3). The NUE of plants for heavy metals had positive relation with plant biomass.
4.1 Plant Growth Performance and Water Use Efficiency
Our findings show that amendment of poultry manure and the biochar of FYM had negative influence on the growth of lettuce but poultry manure when mixed with biochar of wood at both application rates (10% and 15%) and with biochar of FYM at low application rate (10%) caused 2–3-fold increase in aboveground plant biomass and 2–14-fold increase in root biomass. Interestingly, as compared to control treatment, mixture of poultry manure with wood-derived biochar and with FYM-derived biochar significantly increased aboveground plant biomass by ~3 and ~2 fold respectively at 10% amendment rate. Our results are consistent with the finding of Hameeda et al. [36] who also observed enhanced yield of tomato in response to the co-amendment of wood-derived biochar and FYM. Findings of Sun et al. [38–40] also suggested that positive influence of organic fertilizers are enhanced when they are amended in soil with combination of biochars. The soil amended with FYM biochar and with mixture of FYM biochar and poultry manure at higher application rates had significantly higher EC (Supporting Information Tab. S10), which might be the reason for reduced growth of plants possibly because of high concentration of nutrients. Nutrients in high concentration cause osmotic stress in roots and thus affect negatively plant growth [34]. Wood-based biochar have lower concentration of nutrients and act as nutrient capture when applied to soil in mixture with manure [34]. The co-amendment of poultry manure with wood-based biochar might ameliorated the toxic effect of high concentration of nutrients in manure and thus caused a significant increase in biomass of lettuce in our study.
We also tested the influence of poultry manure and the mixture of poultry manure with FYM biochar or wood-based biochar on the growth of another important vegetable crop of Pakistan the spinach. As in lettuce, high amendment rates of poultry manure and biochar of FYM significantly reduced growth, we applied these fertilizers in lower rates, i.e., 3%, 5% and 7%. These amendment rates did not show any response in spinach growth while the mixture of poultry manure with wood-based biochar significantly reduced the growth of spinach as compared to when only poultry manure was applied. This indicates that these amendment rates are not enough to have a positive influence on growth of spinach and the addition of wood-based biochar might have caused nutrient reduction in plants, which caused a reduction in the growth of this crop. The WUE for both crops showed a positive relationship with the aboveground plant biomass. The WUE of lettuce was significantly higher for the treatment of mixture of wood-derived biochar with poultry manure and FYM-derived biochar with the poultry manure as compared to when only poultry manure or wood-derived biochar or FYM-derived biochar were applied to the soil. Such a trend, i.e., biochar + manure mixture positive influence on plant WUE, was however not observed for spinach. This non-consistent trend indicates that these fertilizers have differential influence on the growth performance of these vegetable crops. Crops exhibit variable interaction with their microbiome in response to fertilizer and its application rate, which influence crops growth performance [41]. It is worth further investigation to assess biochar-based organic fertilizer-plant rhizobium interaction in controlling plant growth performance in various crops. Such a study will help understand the crop-specific requirement of biochar-based organic fertilizers regarding their types and application rates.
Poultry manure and FYM in Balochistan province of Pakistan are many-times less expensive than inorganic NPK fertilizer as NPK fertilizer costs approximately 1 US dollar for one kg while FYM costs around 30–45 US dollars per “tone.” Poultry manure if required in small amount (few kg) is even free of cost. These fertilizers when not utilized appropriately, they will be wasted as air and water pollutants [46] with no exception in Balochistan (personal observation). Similarly, major source of wood-derived biochar for Bar B-Q purpose is wood from pruned trees of orchards. The crushed small pieces as leftover of this biochar costs approximately 0.5 US dollars for 1 kg. Agriculture in this arid region can be promoted by utilizing biochar-manure mixture as organic fertilizer. The use of such fertilizer treatments can be proved as cost-efficient and environmental-friendly practice that can potentially also promote agriculture in this arid region of Pakistan.
4.2 Concentration of Heavy Metals (Mn, Cu, Fe, Cd, Pb, Ni, Co)
The NUE of heavy metals in this study was calculated as nutrient efficiency ratio of Baligar et al. [45], which was calculated as plant biomass/concentration of heavy metal in plant biomass [45]. Therefore, high NUE of plant for a given heavy metal (e.g., in our study Mn, Cu, Cd, Pb, Ni or Co) indicates “less” uptake of that heavy metal per unit plant biomass production. As observed the high concentration of heavy metals in poultry manure as compared to wood-based biochar, amendment of these fertilizers showed generally higher concentration of heavy metals in leaves of lettuce as compared to control. However, NUE of lettuce for heavy metals was higher when grown in the soil amended with mixture of poultry manure with wood and FYM-derived biochars. These are the amendments that increased significantly biomass production and WUE of lettuce. Due to limiting funding we could not analyze heavy metals in lettuce for all treatments; however, our observation is in agreement of our previous finding that biochar-based organic fertilizer, which improve biomass production and WUE of plants, also improve NUE of plants for heavy metal absorption [15,36]. For spinach, amendment of mixture of wood-derived biochar with manure reduced concentration of heavy metals. Our results are in agreement to previous findings that wood-derived biochar reduces concentration of heavy metals in plants grown in soil contaminated with high concentration of heavy metals [36]. Wood-derived biochar has low concentration of nutrients. When it is mixed with organic fertilizer that is nutrient-rich it absorbs nutrients from that fertilizer and supply nutrients to plants by acting as slow-release fertilizer [10] and therefore also reduces uptake of heavy metals in plants [47].
In lettuce, concentration of total mineral N and soluble mineral P of soil were significantly and many-fold higher in response to organic amendments as compared to control, indicating that poultry manure, FYM biochar, wood-based biochar and their co-amendments with poultry manure increased concentration of these nutrients. In spinach, concentration of mineral N was higher in soil in response to the amendment of poultry manure and mixture of poultry manure with FYM biochar amended at higher rate (7%) as compared to when mixture of poultry manure and wood-derived biochar was amended at 7% application rate.
Our results indicate that high application rates (10% and 15% or 60 t ha−1 and 90 t ha−1) of poultry manure or FYM biochar, if amended alone reduced significantly the growth of lettuce; however, when these nutrient-rich organic wastes were co-amended (1:1 w/w ratio on air-dry weight bases) with wood-derived biochar, significantly improved biomass production. At low application rates (3%, 5% and 7% or 18 t ha−1, 30 t ha−1 and 42 t ha−1), co-amendment of wood-derived biochar with poultry manure significantly reduced aboveground biomass of lettuce; whereas, co-amendment of FYM biochar with poultry manure at high application rates had no influence as compared to when only poultry manure was applied to soil. This indicates that at low application rates, mixing of poultry manure with wood-derived biochar has no positive influence on this crop, which may be due to at low application rates, these fertilizers do not increase soil nutrients. The mixing of manure-based fertilizers with wood-derived biochar reduced concentration of heavy metals per unit biomass (increased NUE) in both crops. Our findings indicate that higher application rates of poultry manure and FYM biochar need to be done with their mixture with wood-derived biochar. Poultry manure and FYM are many times less expensive than synthetic fertilizer while leftover of wood-derived biochar is available in timber markets of this province (~0.5 US$ per kg). Proper use of biochar-based these organic fertilizers can be cost-efficient for farmers and best waste-management practice.
Acknowledgement: We thank Misbah Manzoor, Department of Plant Sciences, Sardar Bahadur Khan Women’s University, Quetta, Pakistan and Ayeeshaa Masood, Department of Botany, University of Balochistan for their unconditional support in providing laboratory facilities for this study.
Funding Statement: This work is supported by University of Balochistan Research Fund (UBRF-17/026).
Conflicts of Interest: The authors declare that they have no conflicts of interest to report regarding the present study.
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References
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