Quantificação de ácidos humicos e fúlvicos, macro e micronutrientes e razão C/N em fertilizantes orgânicos (em Inglês)

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Communications in Soil Science and Plant Analysis
ISSN: 0010-3624 (Print) 1532-2416 (Online) Journal homepage: http://www.tandfonline.com/loi/lcss20
Quantification of Humic and Fulvic Acids, Macro-
and MicroNutrients and C/N Ratio in Organic
Fertilizers
Ariadne Missono Brondi, Josiane Souza Pereira Daniel, Vitor Xavier Monteiro
de Castro, Alexandre Carvalho Bertoli, Jerusa Simone Garcia & Marcello
Garcia Trevisan
To cite this article: Ariadne Missono Brondi, Josiane Souza Pereira Daniel, Vitor Xavier
Monteiro de Castro, Alexandre Carvalho Bertoli, Jerusa Simone Garcia & Marcello Garcia
Trevisan (2016) Quantification of Humic and Fulvic Acids, Macro- and MicroNutrients and
C/N Ratio in Organic Fertilizers, Communications in Soil Science and Plant Analysis, 47:22,
2506-2513, DOI: 10.1080/00103624.2016.1254791
To link to this article: http://dx.doi.org/10.1080/00103624.2016.1254791
Accepted author version posted online: 29
Nov 2016.
Published online: 29 Nov 2016.
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Quantification of Humic and Fulvic Acids, Macro- and
MicroNutrients and C/N Ratio in Organic Fertilizers
Ariadne Missono Brondi, Josiane Souza Pereira Daniel, Vitor Xavier Monteiro de Castro,
Alexandre Carvalho Bertoli, Jerusa Simone Garcia, and Marcello Garcia Trevisan
Instituto de Química, Universidade Federal de Alfenas – UNIFAL-MG, Alfenas, MG, Brasil
ABSTRACT
Humic substances are themain responsible for soil conditioning and are one of
the major components of the composting product. However, there is still little
information about their composition and mechanisms of activity. This study
aimed to evaluate quantitatively the humic and fulvic fraction by size-exclusion
liquid chromatography and the metal content by atomic absorption spectro-
metry. The chromatographic method used was efficient for humic and fulvic
acids quantitation. Levels of 1.73 ± 0.03 and 1.6 ± 0.5 g kg−1 were found for
chicken manure regarding humic and fulvic acids, respectively, and 5.1 ± 0.9
and 1.2 ± 0.1 g kg−1 for peat. The metal contents indicated the need of a
mineral enrichment, because only the levels of iron reached the minimum
recommended by legislation. It was also observed that evaluated organic
fertilizers were in accordance with the specifications established by legislation
in relation to carbon and total nitrogen and the pathogen levels.
ARTICLE HISTORY
Received 4 May 2016
Accepted 21 September 2016
KEYWORDS
Chicken manure; liquid
chromatography; peat
Introduction
Organic fertilizers are formed by wastes of distinct origins, animal and vegetal and correspond to one of the
most remote material applied on agriculture (Carmo et al. 2013). Among the several types of residues of
animal origin, one highlights the poultry manure (poultry litter) that is considered the main by-product of
aviculture and may be commercialized as natural fertilizer. The use of chicken manure as an organic
fertilizer is very common, acting as source of nutrients for vegetal cultivation and corrector of biological,
chemical and physical conditions of soil. This occurs because manure is rich in phosphorous, nitrogen and
potassium, and so it represents a remarkable source of nutrients and organic matter capable of improving
soil quality of (Witter and Kirchmann 1989), while peat is defined as an organic sediment formed by the
partial decomposition of vegetal matter that presents a high content of humic substances with high capacity
to adsorb metals in transition (Petroni, Pires, and Munita 2000).
In general, organic fertilizer is generated due to the process of fermentation, it means, the activity of
microorganisms on decomposing organic matter and complexing nutrients. The process of organic
matter decomposition is called composting which, when efficiently made, uses organic wastes from
agriculture generating a compound rich in humic substances that are also responsible for chemical,
physical and biological improvements for soil, thus increasing fertility and promoting a balanced
development of plants (Silva 2009a).
The use of organic fertilizers has been growing in Brazil due to low cost, varied composition
and especially to the presence of important nutrients for plants, as well as improvements in
physical and biological quality of soils (Carmo et al. 2013). When searching for inputs less
aggressive to the environment, that allows development of agriculture less dependent on
CONTACT Alexandre Carvalho Bertoli bertolialexandre@yahoo.com.br Instituto de Química, Universidade Federal de
Alfenas - UNIFAL-MG. Av. Jovino Fernandes Sales 2600, Santa Clara, CEP 37130-000, Alfenas, MG, Brasil.
Color versions of one or more of the figures in the article can be found online at www.tandfonline.com/lcss.
© 2016 Taylor & Francis
COMMUNICATIONS IN SOIL SCIENCE AND PLANT ANALYSIS
2016, VOL. 47, NO. 22, 2506–2513
http://dx.doi.org/10.1080/00103624.2016.1254791
http://www.tandfonline.com/lcss
industrialized products, and sometimes imported, there are several products available in the
market. On the other hand, the chemical composition of the organic fertilizers varies according
to the method of preparation and the material of origin. Therefore, an evaluation to guarantee the
quality and agricultural efficiency of these is needed. Furthermore, despite its frequent use in
different cultures, the mechanism of action of the organic fertilizer is not defined, even though
practical results attained were satisfactory (Carmo et al. 2013). Although the inorganic composi-
tion of organic fertilizers sold and produced in the southern region of Minas Gerais, in terms of
micro- and macronutrients, is already known, quantitative information on humic substances is not
described on the labels of this kind of product. Thus, studies that contribute to the development of
analytical methods to determine humic and fulvic acid (HA and FA) concentrations in agricultural
inputs are recommended to scientifically prove their efficiency.
In this sense, the present work aimed to perform the chemical characterization of two organic
fertilizers, mainly in relation to the quantification of HA, FA and metals. The level of their stability
was also evaluated, as well as if the organic fertilizers corresponded to the requirements related to
pathogens. The effect of a chemical treatment on the content of humic and fulvic substances of the
organic fertilizer was also studied by means of the addition of a base.
Experimental
Organic fertilizer samples
Samples of poultry manure and peat, both with about 5 Kg, were provided by the company Café Brasil
Fertilizantes, Alfenas, Minas Gerais. The poultry manure came from the company Ovos Iana in the
municipality of Pouso Alto, Minas Gerais, and identified as a NCTC product, a soil conditioner. The peat
was produced by the company Fertimax Fertilizantes located in the municipality of Boa Esperança,
Minas Gerais. Initially, material, like branches and stones, was previously manually removed. Then,
samples were homogenized,and their quartering wasmade until obtaining approximately 300 g. Samples
were dried in a stove at 60°C until the moisture was lower than 3%. Posteriorly samples were macerated
in porcelain mortar and then sieved using a granulometric sieve of 2-m mesh.
Part of the samples of poultry manure and peat was submitted to chemical treatment in order to
increase the content of HA and FA available to cultivate plants and compare them with the original
samples. This procedure consisted of weighing 100 g of material and adding 30 g of potassium hydroxide
(KOH) (p.a., Vetec Química Fina Ltda). Then, 200 mL of distilled water (H2O) was added. This mixture
was agitated with the aid of a magnetic stirrer for 12 h. After this period, 50.5 mL of phosphoric acid
(H3PO4) (p.a., Vetec Química Fina Ltda) was added with the aim to adjust the pH to values close to
neutrality (6.8 and 6.5 for the manure and peat, respectively). Before adding the base, the manure sample
presented pH of 8.2 and the peat of 4.9. Samples submitted to chemical treatment were named treated
manure and treated peat.
Chemical characterization: content of total organic carbon, nitrogen, copper, iron, manganese
and zinc
The determination of total organic carbon and total nitrogen was performed according to themethodology
recommended by Embrapa (Silva 2009b). Then, sampleswere evaluated in relation to the content of copper,
iron, manganese and zinc. The preparation of samples to determine the content of metallic nutrients was
conducted in open digester block (MA4025, Marconi). Thereunto approximately 0.5 g of the organic
fertilizer samples previously dried in oven at 100°C until constantmass was transferred to specific refractory
tubes of 100 mL. Five mL of nitric acid (HNO3) was added to each tube which was then heated for
10minutes at 95 ± 5°C. After cooling, an additional 5mL of concentrated HNO3 and the tubes were heated
again during 2 h at 95 ± 5°C. After cooling, tubes were sealed with parafilm, and samples rested during
approximately 10 h. After this period, 3 mL of hydrogen peroxide (H2O2) (30% v/v) was added and heated
for 4 h at 120 ± 5°C. After cooling, samples were filtered and the volumemeasured and stored. Triplicates of
COMMUNICATIONS IN SOIL SCIENCE AND PLANT ANALYSIS 2507
each sample and controls (tubes without sample with the addition of all involved reagents and exposed to all
the related conditions for the preparation of samples) were made (USEPA 1996).
Samples were diluted according to the linear range of concentration of the calibration curve for each
metal (0.5–2.5 mg L−1 for copper (Cu), 1.0–7.0 mg L−1 for iron (Fe), 0.5–1.75 mg L−1 for manganese
(Mn) and 0.5–1.75 mg L−1 for zinc (Zn)). The quantification of metallic ions was made through atomic
absorption spectrometer (model AA-7000, Shimadzu) in a flame spraying mode applying a mixture of
air-acetylene and hollow cathode lamps of the respective metals (copper, iron, manganese and zinc).
The conditions of analysis were the same recommended by the manufacturer.
Extraction of the humic and fulvic fraction and quantification by liquid chromatography
The method for extraction of humic substances (HA and FA) adopted in this work was the same
recommended by the International Humic Substances Society (IHSS 2016). To quantify the HA and
FA, a liquid chromatography system UHPLC Ultimate 3000 (Thermo Scientific, California, USA) was
used. The analysis was made at the wave length 254 nm using a BIOSEP-SEC-s2000 column
(300 × 4.6 mm, 5 µm—stationary phase consisting of modified silica, Phenomenex). The mobile
phase used was phosphate buffer (0.028 mol L−1, pH 6.8) with a flow rate of 0.5 mL min−1. The
temperature of the column was held at 25°C, and the run time was 12 min with a 30 µL injection
volume. The chromatographic conditions followed the methodology described by Wu, Evans and
Dillon (2002) with some adaptations.
Approximately 3 mg of extract of HA from each organic fertilizer was dissolved in 1 mL of
sodium hydroxide (NaOH) 0.1 mol L−1 using 10-mL flasks. This material remained resting during
24 h. After this period, the volume was adjusted with buffer solution of phosphate (pH 6.8). Before
the chromatographic analysis, samples were filtered using a 0.22-µm polyvinylidene fluoride mem-
brane (PVDF, Millipore).
Adequate volumes (from 0.5 to 5 mL) of the fractions of FA extracts from each organic fertilizer
were transferred to the flask of 10 mL, which was completed with buffer solution of phosphate
0.028 mol L−1 and the pH adjusted for 6.8. In both analyses, made in triplicates, 30 µL was injected.
The standard solutions of HA and FA used to build the calibration curves of the chromatographic
analysis were prepared from materials recommended by the IHSS called Pahokee Peat Humic acid
(code 1S103H) and Pahokee Peat Fulvic acid (code 2S103F). The calibration curves were prepared
from 15 to 150 µg mL−1 for HA and 50 and 280 µg mL−1 for the FA.
Microbiological analyses
Samples of organic fertilizers were submitted to microbiological analyses to verify the presence of
helminth eggs, thermotolerant coliforms and Salmonella sp according to a procedure described by
Silva (2010). Such analyses were made to check whether the organic fertilizers were in accordance
with the requirements of the Normative Instruction n. 27 from 2006 of the Ministry of Agriculture,
Livestock and Supply (MAPA 2006).
Statistical analyses of data
All data of quantification of humic and fulvic substances and metals were presented as mean values
followed by the respective standard deviations obtained from at least three repetitions. Differences
among samples were determined using ANOVA at the significance level of 0.05 according to the
F-test to verify the difference of HA and FA content in manure and peat, before and after the treatment,
and the t-test was used to compare the results obtained with those recommended by the law.
2508 A. M. BRONDI ET AL.
Results and discussion
Content of total C and N, macro- and micronutrients
The contents of total carbon and nitrogen found in the analyses are available in Table 1 and
presented adequate reproducibility due to the low standard deviation. All evaluated samples were
in accordance with the minimum requirements established by MAPA in the Normative Instruction
n. 25 from 2009 which established a minimum content of organic carbon of 20 and 15% for manure
and peat, respectively, and for total nitrogen 1 and 0.5% are required for the same material (MAPA
2009). Therefore, both the organic fertilizer from poultry wastes and peat have potential to be used
in agriculture. It is important to mention that the manure sample presented both for carbon and
nitrogen content of approximately 16 and 75%, respectively, when was superior when compared to
peat the values. Furthermore, the addition of KOH and H3PO4 did not alter the content of organic
carbon and total nitrogen of the samples, as expected. We highlight that the content of organic
carbon is independent of the oxidation state of organic matter and does not consider other organic
elements (Silva 2009b).
According to Inácio and Miller (2009), a C/N ratio greater than 20:1 may indicate that the
material is not properly stabilized that the material still can be liable to suffer the strong action of
microorganisms when in contact with the soil. Thus, results pointed that the organic fertilizers
evaluated in the present work have adequate stability since they presented a C/N ratio lower than 18,
an important parameter of quality for use in soils.
Lourenço (2013) found a carbon to nitrogen (C/N) ratio on the order of 10 for poultry litter
constituted by corn stover, sugarcane bagasse, pasture straw, sand and Pinus acicula. In the present
work, a C/N ratio of 11.8 was found for the same material.
The Normative Instruction n. 25 from 2009 also recommends the minimum requirements related
to secondary macronutrients and micronutrient in organic fertilizers (MAPA 2009). They are
recommended minimum levels of 0.05,0.2, 0.05 and 0.1% for Cu, Fe, Mn and Zn, respectively,
for application in the soil. It is recommended Table 2 the content of metallic ions of organic
fertilizers samples is presented.
Manure, both the treated and in natural, had a high content of iron, which was about eight times
greater than the minimum required by MAPA (2009) in the Normative Instruction n. 25 from 2009,
and zinc, approximately two times the minimum recommended level. However, the content of
copper and manganese did not meet the previously mentioned normative. Peat, before and after
treatment, presented approximately four times the minimum content of iron; however, it did not
Table 1. Content of total carbon and nitrogen in the organic fertilizer samples.
Sample Total Organic Carbon (%)* Total Nitrogen (%)* C/N*
Manute 21.3 ± 0.4 1.83 ± 0.01 11.6 ± 0.4
Treated manure 21.5 ± 0.2 1.82 ± 0.03 11.8 ± 0.2
Peat 18.3 ± 0.2 1.04 ± 0.05 17.6 ± 0.2
Treated peat 18.1 ± 0.5 1.07 ± 0.03 16.9 ± 0.5
*Conc. ± I.C. (95% of confidence, n = 3).
Table 2. Concentration of metallic ions in the organic fertilizers evaluated through atomic absorption spectrometry.
Concentration (mg g−1)*
Samples Cu2+ Fe2+ Fe3+ Mn2+ Zn2+
Minimum MAPA 0.5ª** 2ª** 0.5ª** 1ª**
Manure 0.28 ± 0.04b** 16.4 ± 0.3b** 0.34 ± 0.04b** 2.6 ± 0.3b**
Treated manure 0.23 ± 0.01c** 12.5 ± 0.2b** 0.20 ± 0.04c** 2.1 ± 0.3c**
Peat 0.06 ± 0.02d** 8.22 ± 0.07c** 0.03 ± 0.01d** 0.30 ± 0.01d**
Treated peat 0.02 ± 0.01e** 5.90 ± 0.07d** 0.01 ± 0.00e** 0.22 ± 0.02e**
*Conc. ± I.C. (95% of confidence, n = 3); **t-test (95% of confidence, n = 3, tcrit = 2.920).
COMMUNICATIONS IN SOIL SCIENCE AND PLANT ANALYSIS 2509
meet the minimum recommended levels of the other metallic ions. These values may vary according
to the feeding and metabolism of chickens, in the case of manure, or in the case of the plant material
that was used to produce the peat. Therefore, in some cases, mineral enrichment is required. This is a
procedure commonly adopted for organic fertilizers.
Analysis of the humic and fulvic fraction by liquid chromatography
After the fractionation of humic substances, according to the procedure recommended by IHSS, samples
were analyzed by HPLC (high performance liquid chromatography). However, initially the standards of
HA and FA were analyzed to obtain the calibration curve based on peak area. It is important to mention
that the mobile phase used was also evaluated, and it did not present any peak during the analysis. The
obtained coefficients of correlation (R2) were 0.9983 and 0.9848 for the standards of HA and FA,
respectively. Figure 1 presents chromatographic peaks showing retention time of 7.1 and 7.4 minutes,
corresponding to the humic and fulvic fractions, respectively, present in the standards. In the samples of
organic fertilizers, there was a small displacement (0.2 min) for superior retention times.
Although all the chromatograms exhibited similar shapes, the humic fraction presented retention
time a little inferior because of the greater mean molar mass and wider peak due to the high complexity
also expressed by the chemical, physical and structural poly-dispersion when comparing with the fulvic
fraction (Figure 1). This behavior is consistent with other studies available where the humic fraction
presents lower retention time when compared to the fulvic fraction, since it possesses a higher mean
molar mass. Furthermore, the chromatographic peak corresponding to the humic fraction is more
diffuse (Hur, Williams, and Schlautman 2006; Maia, Piccolo, and Mangrich 2008).
In the work of Hur, Williams, and Schlautman (2006), they evaluated the standards Suwannee River
Fulvic acid and Aldrich Humic acid and also the presence of HA and FA in natural samples of water
using size-exclusion liquid chromatography. However, the obtained retention times were 10 min for HA
and 13.5 min for FA, and these data are related to the apparent weigh and the mean number of molecular
weight of humic substances.
Wu, Evans and Dillon (2002) also quantified HA and FA in natural samples of water, and they
obtained retention times of 11.3 and 12.4 min, respectively, for the standard of HA and FA (Wu 2007).
These values differ from what was found in the present work, but it is important to highlight that the
standards (Suwannee River Fulvic acid and Aldrich Humic acid) and the column (stationary phase
comprising hydroxylated polymethacrylate with a diameter of 7.8 mm) used in the work of Wu, Evans
and Dillon (2002) were different, what justifies such behavior (Wu 2007).
Humic acid
0 2 4 6 8 10 12
0
20
40
60
80
100
 Standard solution 150 µg/mL
 Manure
 Treated manure
 Peat
 Treated peat
In
te
ns
it
y 
(A
. U
.)
Time (min)
0 2 4 6 8 10 12
0
50
100
150
200
250
300
350
400
In
te
ns
it
y 
(A
. U
.)
Time (min)
 Standard solution 280 µg/mL
 Manure
 Treated manure
 Peat
 Treated peat
Fulvic acid
Figure 1. Chromatographic profile of humic and fulvic acids present in the standards and samples of organic fertilizers obtained
from mobile phase constituted by phosphate buffer (pH 6.8) at 0.5 mL min−1.
2510 A. M. BRONDI ET AL.
Maia, Piccolo and Mangrich (2008) analyzed qualitatively sawdust sludge and grounded paper
submitted to a composting process for 15 days in a bioreactor for the distribution of molecular sizes of
the HAs. Analyses were made by size-exclusion liquid chromatography applying a column called
Polysep GFC-P3000 (600 × 7.8 mm) and using 100 µL of sample for the injection. The mobile phase
was phosphate buffer at 0.1 mol L−1 (pH 7) at a flow of 0.6 mLmin−1. Under these conditions, the HAs
eluted in 25 min. Then, it is important to emphasize that the quantification method developed in the
present work may be considered advantageous since the HA and FA presented lower elution times
(approximately 7 min and 30 sec).
The content of HAs in the organic fertilizer samples processed by adding KOH andH3PO4 was higher
when compared to the samples without treatment (Table 3). This fact was mainly caused by the presence
of KOH. Studies point that an increase of KOH concentration provides a greater extraction of humic
substances. This is explained by the thermodynamic characteristics of potassium ions (radius of hydra-
tion and ion mobility) that influence the solvation process of humic substances ionized groups and,
consequently, the yield of extraction (Romarís-Hortas, Moreda-Piñeiro, and Bermejo-Barrera 2007;
Rosa, Rocha, and Furlan 2000). For example, one may cite the work of Rosa, Rocha and Furlan (2000)
who affirmed that the increase on the base concentration (from 0.1 to 1 mol L−1) lead to the decrease in
the HA extraction and increase in FA extraction for the peat sample collected in the municipality of
Ribeirão Preto, São Paulo. Considering the ratio material/solution of KOH at 1 mol L−1 1:10 (m/v),
0.01 mol of base was applied for each 1 g of peat. In the present work, this tendency was not observed.
This may have occurred due to the higher concentration of KOH that was applied during the treatment
(approximately 0.018 mol for each 1 g of organic fertilizer). Furthermore, increasing the pH there is the
increase of solubility of HA because of its ionization.
The analyzed organic fertilizers presented more HA in relation to FA, and in the treated samples, a
significant increase in the HA concentration was observed. It was possible to note an increase of 2.5 times
in the samples of manure and peat. According to Silva (2009a), the predominance of HAs in relation to
the FA in the final of composting indicates an adequate humification of the evaluated organic fertilizers.
On the other hand, it was noted that the concentration of FA in themanure sample was statistically equal,
according to the F-test, even with the addition of KOH. But in the peat sample, the base provided an
increase of 3.6 times on the content of FA.
Benites (2004) related that in a final product of grass composting after 82 days presented 53.4 and
39.2 g kg−1 of HA and FA, respectively.These values are much higher than what was found in the present
work. However, the conditions of extraction are not those recommended by IHSS, a simplified and low
cost procedure was used and the determination of substances was conducted by through titration with
ferrous ammonium sulfate.
We highlight the difficult in comparing the content of HA and FA, because the methodologies of
extraction and determination of these substances, mainly in solid samples, are very different. This is a
complicating factor since several types of organic fertilizers have appeared in the market, which need to
be evaluated in terms of the typical composition that determines the agricultural value, including the
content of humic substances.
It is important to mention that the addition of compounds containing humic substances may stimulate
the growth of plants. In the work developed by Marrocos, they evaluated the performance of melon
cultivated via fertigation with organic fertilizers from cattle and poultry manure in different dosages and in
Table 3. Concentration of humic acid (HA) and fulvic acid (FA) obtained through HPLC.
Sample HA (g kg−1)* FA (g kg−1)*
Manure 1.73 ± 0.03ª** 1.6 ± 0.5ª**
Treated manure 4.1 ± 0.1b** 1.9 ± 0.1ª**
Peat 5.1 ± 0.9c** 1.2 ± 0.1b**
Treated peat 12.9 ± 0.1d** 4.4 ± 0.2c**
*Conc. ± I.C. (95% of confidence, n = 3); **F-test (95% of confidence, n = 3, Fcrit = 0.0526); *** Values in the columns followed by
the same letter did not present significant difference at the mentioned confidence interval.
COMMUNICATIONS IN SOIL SCIENCE AND PLANT ANALYSIS 2511
distinct times of decomposition (Carmo 2013). The greatest productivity of fruits was observed when the
organic fertilizer with 25% potassium was used. Characteristics of quality like total acidity and bark
thickness presented an effect due to the use of different dosages of organic fertilizers. In relation to the
meanmass of commercial fruit, it was observed that the organic fertilizationwith poultrymanure presented
results significantly superior to cattle manure in dosages of 25% and 100%. Witter and Kirchmann (1989),
also evaluated the effects of the presence of organic fertilizer of cattle manure on the development of
American-type of and fresh lettuce. It was verified that the application of organic fertilizer in the dosage, 2%
allowed and increase of 67% of fresh material of the commercial part of American lettuce and 56% of dried
material when compared to the control. On the other hand, the application of organic fertilizer in the
culture of curly lettuce did not provide significant differences in relation to the phytotechnical parameters
(height, fresh mass of the aerial part, number of leaves and circumference of the head), thus masking its
nutritional effect due to the lack of fertility in the applied soil.
Analyses of pathogens
The organic fertilizer was in accordancewith the limits recommended by the Normative Instruction n. 27
from 2006 for all the conducted microbiological analyses. The tests for thermotolerant coliforms,
Salmonella sp and helminth eggs in all samples had negative results (MAPA 2006).
Once detecting pathogenic bacteria from various samples in environmental, food and agricultural
fields has been of great importance because pathogenic bacteria infection is found not only in foods, but
also in soil, water and air environments (Baek 2015).
Conclusions
The ratio C/N indicated that the evaluated organic fertilizers have adequate stability to be applied in
soils. The treatment with KOH and H3PO4 did not result in alterations of this parameter. Only iron
was in accordance with the recommended by MAPA in the Normative Instruction n. 25 from 2009
(MAPA 2009). On the other hand, the content of zinc, copper and manganese ions indicated the
need of mineral enrichment both for the organic fertilizer without treatment with KOH and H3PO4
and the treated one.
According to the results, the peat presents a concentration of HA about five times higher than the
poultry manure and concentration of FA statistically equal, according to the F-test.
The treatment with addition of KOH and H3PO4 led to an increase of about 2.5 times the
concentration of HA in the samples of manure and peat. On the other hand, there was an increase of
3.6 times in the content of FA only in the peat sample.
Funding
Authors thank exclusively the Fundação de Amparo de Pesquisa do Estado de Minas Gerais—FAPEMIG (Process
APQ-04835-10) for the financial support. We also thank Frederico Araújo Leite and Maria Tais Buzzo Gomes of Café
Brasil Fertilizantes, Alfenas (MG), Brasil.
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COMMUNICATIONS IN SOIL SCIENCE AND PLANT ANALYSIS 2513
http://dx.doi.org/10.1111/j.1757-1707.2012.01199.x
http://dx.doi.org/10.1016/j.chemosphere.2005.08.069
http://dx.doi.org/10.1016/j.chemosphere.2005.08.069
http://www.humicsubstances.org/soilhafa.html
http://dx.doi.org/10.1590/S0100-06832013000200017
http://dx.doi.org/10.1016/j.chemosphere.2008.07.069
http://extranet.agricultura.gov.br/sislegisconsulta/consultarLegislacao.do?operacao=visualizar%26id=16951
http://extranet.agricultura.gov.br/sislegisconsulta/consultarLegislacao.do?operacao=visualizar%26id=16951
http://www.agricultura.gov.br/vegetal/fertilizantes/legislacao
http://www.agricultura.gov.br/vegetal/fertilizantes/legislacao
http://dx.doi.org/10.1590/S0100-40422000000400009
http://dx.doi.org/10.1016/j.aca.2007.09.022
http://dx.doi.org/10.1590/S0100-40422000000400008
http://www3.epa.gov/epawaste/hazard/testmethods/sw846/pdfs/3050b.pdf
http://dx.doi.org/10.1007/BF02220694
http://dx.doi.org/10.1016/j.apgeochem.2007.03.043
http://dx.doi.org/10.1016/S0003-2670(02)00476-2
	Abstract
	Introduction
	Experimental
	Organic fertilizer samples
	Chemical characterization: content of total organic carbon, nitrogen, copper, iron, manganese and zinc
	Extraction of the humic and fulvic fraction and quantification by liquid chromatography
	Microbiological analyses
	Statistical analyses of data
	Results and discussion
	Content of total C and N, macro- and micronutrients
	Analysis of the humic and fulvic fraction by liquid chromatography
	Analyses of pathogens
	Conclusions
	Funding
	References