Journal of the Air & Waste Management Association

ISSN: 1096-2247 (Print) 2162-2906 (Online) Journal homepage: http://www.tandfonline.com/loi/uawm20

Component optimization of dairy manure

vermicompost, straw, and peat in seedling
compressed substrates using simplex centroid
design

Longyuan Yang, Hongliang Cao, Qiaoxia Yuan, Shuai Luo & Zhigang Liu

To cite this article: Longyuan Yang, Hongliang Cao, Qiaoxia Yuan, Shuai Luo & Zhigang Liu
(2017): Component optimization of dairy manure vermicompost, straw, and peat in seedling
compressed substrates using simplex centroid design, Journal of the Air & Waste Management
Association, DOI: 10.1080/10962247.2017.1368736

To link to this article: http://dx.doi.org/10.1080/10962247.2017.1368736

Accepted author version posted online: 22

Aug 2017.

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 Component optimization of dairy manure vermicompost,
straw, and peat in seedling compressed substrates using
simplex centroid design
Longyuan Yang 1, Hongliang Cao1,2, Qiaoxia Yuan *1,2, Shuai Luo1, Zhigang Liu1,2
1
College of Engineering, Huazhong Agricultural University, No. 1, Shizishan Street,
Hongshan District, Wuhan 430070, PR China;

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2
The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070,
PR China.

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*Please address correspondence Qiaoxia Yuan, E-mail address: yc1group@126.com

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Abstract
Vermicomposting is a promising method to disposal dairy manures, and the dairy
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manure vermicompost (DMV) to replace expensive peat is high-value in the
application of seedling compressed substrates. In this research, three main
components, DMV, straw, and peat are conducted in the compressed substrates, and
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the effect of individual component and the corresponding optimal ratio for the
seedling production are significant. To address these issues, the simplex-centroid
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experimental mixture design is employed, and the cucumber seedling experiment is

conducted to evaluate the compressed substrates. Results demonstrated that the
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mechanical strength and physicochemical properties of compressed substrates for

cucumber seedling can be well satisfied with suitable mixture ratio of the components.
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Moreover, and peat) could be determined at 0.5917: 0.1608: 0.2475 when the weight
coefficients of the three parameters (shoot length, root dry weight and aboveground
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dry weight) were 1: 1: 1. For different purpose, the optimum ratio can be little
changed on the base of different weight coefficient.

Keywords: compressed substrates; vermicompost; cucumber seedlings

Implication. Compressed substrate is lump and has certain mechanical strength,

produced by application of mechanical pressure to the seedling substrates. It will not
 harm seedlings when bedding out the seedlings, since the compressed substrate and
seedling are bedded out together. However, there is no one using the vermicompost
and agricultural wastes components of compressed substrate for vegetable seedling
production before. Thus, it is important to understand the effect of individual
component to seedling production, and to determine the optimal ratio of components.

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INTRODUCTION

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With the fast development of dairy feeding operations, the production of dairy

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manures is dramatically increasing, which lead to increasing the severity of

environmental problems (Cao et al. 2014, Saunders et al. 2012, Venglovsky, Sasakova,

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and Placha 2009). These problems cannot be neglected. Dairy manure is high in
organic matters and contains nutrients essential for crop production, which is effective
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and popular as fertilizer (Cao, Xin, and Yuan 2016, Kapkiyai et al. 1999). However,
the direct use of dairy manure in an unmodified form for seedling production is not
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suitable, as untreated dairy manure is a significant source of ammonia volatilization,
which can lead to an opposed effect on crop growth (Chadwick 2005). Moreover,
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dairy manure can also be an important source of potentially dangerous pathogens and
nitrates that may be transported to the surface and ground water (Uzoma et al. 2011).
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Composting is a widely used disposal technology, which can produce a stabilized

product (Cao, Xin, and Yuan 2016, Hao et al. 2001). However, C and N losses during
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composting not only reduce the agronomic value of compost as fertilizer, but also
contribute to emissions of greenhouse gas, such as CO2, CH4, and N2O (Brown,
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Wagner-Riddle, and Thurtell 2000, Hao et al. 2001). Recently, vermicomposting is a

new and promising method to disposal dairy manures. During the process, the dairy
manure is digested by earthworms, and the earthworm manure, vermicompost, is a
high-quality fertilizer with increased availability of nutrients and improved
physicochemical and microbiological properties, compared with the traditional
composts (Atiyeh et al. 2001, Ding et al. 2016, Orozco et al. 1996). More importantly,
vermicomposts have similar physicochemical characteristics with peat, such as bulk
 density, electrical conductivity, salt content, and pH value (Jin and Chang 2011). In
China, peat is widely used as the main substrate component for the production of
seedlings (Bustamante et al. 2008, Garcia-Gomez, Szmidt, and Roig 2002). However,
the resource of peat is limited and non-renewable. Therefore, the peat is rare and of
high cost for seedling production. The dairy manure vermicompost can replace peat
with a high ratio, even 100%, for seedling production of cucumber (Hashemimajd et
al. 2004). Therefore, dairy manure vermicomposting is a kind of high value-added

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seedling substrates.
However, during the seedling experiment, seedling production was conducted

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with plastic plug, where the substrates, such as peats and vermicomposts, were used in

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the form of incompact. In actual production, a large number of plastic plugs are used,
which resulted in serious resources waste and environmental pollution. Moreover, this
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kind of seedling method, plug-seedling, has some other disadvantages. For instance,
the process is complex, including substrate preparation, batching, loading, seedling
management, removing the plastic plug, bedding out, and so on. Moreover, the poor
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air permeability is also an obvious shortcoming, where the absorption of the air and
water by the seedlings is difficult (Kostopoulou et al. 2011). In addition, removing the
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seedlings from the plastic plug is also difficult and would injure the root of seedlings,
which is harmful for the subsequent seedling stage (Menge et al. 1978). Therefore, the
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plug-seedling method is gradually eliminated, and a new integrative method,

compressed substrate (CS), is increasingly popular for seedling production. CS is
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lump and has certain mechanical strength, produced by application of mechanical

pressure to the seedling substrates, such as peats, vermicomposts and so on. In this
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case, during seedling production the plastic plug is not used, which is helpful for
saving resources and protecting environment. More importantly, it will not harm
seedlings when bedding out the seedlings, since the CS and seedling are bedded out
together. Thus, the subsequent seedling stage is very short and insignificant (Renison,
Cingolani, and Schinner 2002).
Although, CS is more suitable for seedlings than incompact substrates, the former
has stricter demands for substrates than the latter. Besides good chemical properties
 (such as pH for potential of hydrogen, EC for electric conductivity, N for nitrogen, P
for phosphorus, K for potassium, trace elements), CS should also have favorable
physical characteristics, such as mechanical strength, density, water availability, air
capacity, porosity, and expansion rate. In this context, it is important to address the
components of substrates as well as their ratios. Considering physical characteristics,
Dairy Manure Vermicompost (DMV) has good density, water availability, air capacity,
and porosity, which are beneficial for CS. However, its cohesiveness is not very well,

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worse than peat, which makes against the molding of CS. Therefore, in this research,
the components of CS still consider partial peats. Moreover, straw is also added into

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the CS, since straw has very well cohesiveness. In this way, we can reduce the ratio of

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peat, which is helpful to reduce the cost of CS. In addition, a small quantity of
super-absorbent polymer is also added to enhance the porosity, water availability, and
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expansion rate of CS.
This research aims to determine the effect of individual component to seedling
production, and to identify the optimal ratio of components. The simplex-centroid
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experimental mixture design is employed to study relationships among proportions of

different variables and responses which not only establish the surface model of
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continuous variables, estimating every element in the mixture and their interactions,
but also optimize the component elements according to the target (Muteki, MacGregor,
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and Ueda 2007). Besides, the cucumber seedling experiment is conducted to evaluate
the CS with different component ratios. It should be mentioned that this work may be
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is the first study to address DMV for CS seedling application, which provides a useful
reference for the clean and value-added process of dairy manures as well as other
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organic wastes.
 MATERIALS AND METHODS

Raw materials

DMV was produced in aerated piles of worm-inoculated dairy manure at the

earthworm farm in Hubei province, China. The vermicompost was air-dried, smashed

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by a hammer mill, and sieved through a 5mm mesh screen before addition to

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compressed substrates. The used peat (Fafard brand) was imported from Canada.

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The rice straw was collected from farm households around Huazhong Agricultural
University (Wuhan City, Hubei Province, PR China), milled and smashed into

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filiform straw through a 2mm mesh screen (length ≤ 1.5 cm). The used
super-absorbent polymer (Renqiu city XinGuang chemical industry Co., LTD,China)
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had a water absorption rate of 100g/g and expansion rate of 5(v/v). The base
physicochemical characteristics of these raw materials are listed in Table 1.
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Experimental design
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Scheffe (1963) proposed the simplex centroid design for optimal experimental
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design of testing the effects of individual components on the overall quality of mixtures.
DMV, smashed straw and peat were selected for the optimization of compressed
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substrate to optimize the efficiency of seedling production and to investigate if these

components have any synergistic or antagonistic effects when blended. To use DMV as
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much as possible, its rate in substrates was controlled above 50% by volume.
We employed a three-component, augmented simplex-centroid design in which
components are tested in a concentration gradient (by volume) across seven mixtures: 0
(0%), 1/6 (16.7%), 1/4 (25.0%), 1/2 (50.0%), 2/3 (66.7%), 3/4 (75.0%) and 1 (100.0%).
A ternary phase diagram of the 7-point test is obtained (Figure 1). The matrix of the
simple centroid mixture design and response values for the compressed substrate
breakage rates before the growth of the seedlings in compressed substrates (BRB), the
 compressed substrate breakage rates after the growth of the seedlings in compressed
substrates (BRA), shoot length, root dry weight and aboveground dry weight are listed
in Table 2. A 3g of super-absorbent polymer (SAP, Cross-linking Copolymerization of
Acrylic Acid) was added per liter of mixture to improve the expansion of compressed
substrates.
Linear, quadratic or special cubic regression models (see Eq.1) were used for
variations of all responses as a function of significant interaction effects (p < 0.05)

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between the proportions, with acceptable determination coefficients (R2> 0.90).

Yi   i X i  ij X i X j  ijk X i X j X k (1)

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where Yi is the predicted response,  i is the regression coefficient for each linear

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effect term,  ij and ijk are binary and ternary inter-action effect terms and X i , X j
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and X k are the coded independent variables. (In this work, X 1 represents the DMV,

X 2 represents the rice straw, X 3 represents the peat)

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Compression of substrates and management of the experiment

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Materials were mixed according to the listed proportions in Table 2, with added
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SAP. Subsequently, 40g of each mixture were compressed at 10kN pressure in a tubular
mold (inner diameter 50mm) with a hydraulic press (WE-100B brand, made in China
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by Zhejiang Jingyuan Machinery Equipment Co., LTD). The compressed substrate

samples are shown in Figure 2(a). The plant growth experiment was carried out inside
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a greenhouse at Huazhong Agricultural University (Wuhan City, Hubei Province, PR

China) for 20 days in May 2016.The monthly average temperature during the day and
night was 26°C and 17°C, respectively. The experimental process is as follows:
cucumber seeds were soaked in warm water (50°C) for 20 minutes and moved to wet
gauze on a tray, then the tray immediately are placed in a 30°C culture box with no
illumination. The gauze was moistened every 6 hours. Cucumber seedlings with similar
growth were sowed in the holes of pre-watered. The compressed substrates after
 watering are illustrated in Figure 2(b). Micro-spray irrigation with tap water (no
fertilizer) was used throughout the seedling stage (Figure 2(c)). Figure 2(d) shows the
seedling growth after 20 days. The number of compressed substrate in each formulation
treatment was 20, and each formulation treatment was conducted with triplicates.

Measurement of compressed substrate breakage and analytical methods of

physicochemical properties

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In this study compressed substrate breakage rates were quantified to evaluate the

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effect of compression BRB and BRA. To determine breakage rates, the compressed

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substrates were weighed as m1. After dropping the individual compressed substrates
from 1 meter above the ground, the biggest broken compressed substrate was chosen
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and weighed again as m2. Thus, the breakage rate before planting the cucumber
seedlings (BRB) was calculated as follows:
(m1 −m2 )
%BR B = × 100
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(2)
m1

After growing the cucumber seedlings, the number of broken compressed substrates
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(which were broken by watering and not convenient for transplant as a whole) was
calculated as A1, with the total number of compressed substrates for each mixture as A2.
The breakage rate after seedling growth (BRA) was calculated as follows:
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A
%BR A = A1 × 100 (3)
2
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Electrical conductivity (EC) and pH were measured in the water extract of 1:5 by
volume of each mixture, according to Johnson (1980). Physical properties of the
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mixtures were determined according to methods of Inbar, Hadar, and Chen (1993) for
water availability, air capacity, total porosity and bulk density. (Bulk density = dry
weight/volume; particle density = 1/[% organic matter/(100×1.55)+ % ash/(100×2.65);
total porosity =(1- bulk density/particle density) ×100%;water availability = [(wet
weight – dry weight)/volume] × 100%; air capacity = (total porosity - water
availability) ).
After the seedling growth, aerial parts and roots of cucumber seedlings were dried
 at 105°C for 15 minutes and then dried at 65°C as previously reported to measuring dry
weight. Sub-samples (3 replications per treatment) of substrates and seedlings were
screened by 0.25mm sieve and taken for chemical analysis. After vitriol digestion of the
sub-samples, nitrogen and phosphorus were determined by Flow Injection Analysis and
all other nutrients were determined by Atomic Absorption Spectrophotometry. Data
were subjected to ANOVA analysis (with SAS® 9.1 software) followed by Duncan’s
multiple range test (p =0.05). The coefficient of determination R2 and F test (analysis

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of variance-ANOVA) were used to verify the quality of fit of the models.

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RESULTS AND DISCUSSION

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Physical properties
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Table 3 presented the physical properties analyses of different compressed
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substrates. . It can be found that the bulk density of compressed substrates, after the
incorporation of 0% to 50% smashed straw into DMV, reduced by 0-0.08 g/cm3, while
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the bulk density of compressed substrates with DMV as the only added waste
component were the highest (0.49 g/cm3) of all tested formulations.
Increasing addition of smashed straw (0% to 50%) into DMV resulted in no
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distinct reduction in bulk density, total porosity and air capacity. On the contrary,
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increasing 50% peat resulted in significant reduction in bulk density, but did cause an
increase in total porosity and a change in pore space distribution within the compressed
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substrates that raised water availability and air capacity. Atiyeh et al. (2001) reported
similar results from substrates composed of peat with different concentrations of pig
manure vermicompost. The water availability of all compressed substrates (except TS1
and TS4) was in the optimum range, while the air capacity of all mixtures was
unacceptable due to high levels of variability. However, according to Bragg and
Chambers (1988) and Handreck and Black (2002), an air capacity between 7% and 10%
was acceptable as long as watering was carefully managed. The expansion rate after
 watering correlates positively (Pearson coefficient, the value of sig was 0.023<0.05),
with total porosity of compressed substrates and higher expansion rate, which is more
favorable for transportation and plant growth because these compressed substrates
could be compressed to a smaller volume for transportation and a high expansion after
watering, where it had a higher total porosity for plant growth. Adding of smashed
straw or peat into DMV, both increased the expansion rates of compressed substrates,
with a more pronounced increase in peat treatments.

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Chemical properties

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Results in Table 4 show that the pH values of all compressed substrates were
within the optimal range for plant growth (5.0-6.5) according to Abad, Noguera, and
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Burés (2001). Upon incorporation of smashed straw or peat into DMV, the pH of the
compressed substrates decreased progressively with increasing the proportions of
smashed straw or peat. . The highest EC value (2.04±0.02 mS/cm) was given by 100%
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dairy manure vermicompost (with SAP) as a compressed substrate. The EC values of

compressed substrates decreased significantly with the addition of smashed straw or
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peat, though EC values of all formulations when it exceeds 1-3 mS/cm (Gajdoš 1997).
Also, the macronutrients of compressed substrates are listed in Table 4. By adding
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50% smashed straw into DMV, the contents of total phosphorus (TP), total potassium
(TK), and manganese (Mn) were increased significantly compared to DMV-only
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compressed substrate, whereas the rest of elements (Table 4) were unchanged across
treatments. Upon incorporation of 50% peat (by volume) the concentrations of TP,
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calcium (Ca), and magnesium (Mg) increased commensurately with adding of peat.
Meanwhile, levels of Mn were lowered significantly. Concentrations of other elements
were stable regardless of peat content. The heavy metals in all compressed substrates
were significantly lower than the stated limits by Abad et al. (1993). These limits, in
terms of mg kg-1 of dry weight, are as follows: copper (Cu), 500; zinc (Zn), 1500. When
the three materials were mixed (TS6 and TS7), all tested elements in the compressed
substrates were relatively abundant and balanced.
 Model fitting and regression analysis

The response data based on the independent variables was obtained from the
experiments and recorded in Table 2. The experiments were conducted with
triplicates and found that in almost all cases there exists good agreement between the
original and triplicates. All the independent and response variables were fitted to

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linear, quadratic or special cubic models as shown in Table 5, the predicted regression
equations represent the models for BRB, BRA, shoot length, root dry weight and

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aboveground dry weight. Among the independent variables, the peat ( x3 ) exerted the

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highest significant effect on the BRA, shoot length and aboveground dry weight,

followed by SS ( x2 ) and DMV ( x1 ). The peat ( x3 ) showed the highest positive

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significant effect on BRA, shoot length and aboveground dry weight, whereas it
simultaneously showed the highest negative significant effect on root dry weight.
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Table 6 contains the models, corresponding R2 and adjusted R2 of the regression
equations for the responses as well as the corresponding F-ratio and p-values for each
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term in the predicted regression equations. All models gave R2 higher than 0.87
indicating a very good fitting. The best fit was obtained by the quadratic or special
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cubic models for BRB, BRA, shoot length, root dry weight and aboveground dry weight.
Comparison between the different models according to the F-test values showed that
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the calculated F-test values were less than the critical F-test values at 95% confidence
level, indicating that all models have statistical significant and can be used for
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predictive purposes in the simultaneous reduction the breakage (BRB and BRA) of
compressed substrates and production of strong cucumber seedlings.
 Effect of mixture on breakage rate of compressed substrates

The compressed substrates often needed to be moved, resulting in broken substrate,

therefore, the BRB of compressed substrates had a negative correlation with movement
of compressed substrates, while the lower breakage rate showed the high quality of
compressed substrates before the seedling stage. The variation in breakage rates

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between compressed substrates made with combinations of DMV, straw and peat are
shown using mixture contour plots in which each factor is represented in a corner of an

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equilateral triangle; each point within this triangle refers to a different proportion of

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components in the mixture (Figure 3(a)). It was observed that the minimum response
variables were located toward the side of triangle having mixtures of DMV and
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smashed straw or DMV and peat as the vertices. This indicates that, to some extent,
smashed straw and peat may be added to decrease the breakage rate of compressed
substrates. The filiform shape of smashed straw and peat may act as reinforcement to
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increase the structural stability of the compressed substrate.

Figure 3(b) showed the breakage rate response variables, BRA, for mixtures TS1,
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TS2 and TS5 after 20 days of growth by the cucumber seedlings. The observation of
breakage in TS1, TS2 and TS5 (as compared to no breakage in other samples) is
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possibly due to the lack of smashed straw amendment in TS1, TS2 and TS5 or the high
expansion of peat. These data indicate that in order to minimize the breakage rate
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response variable, smashed straw should be included in the compressed substrate

formulation.
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Effects of mixture on stem diameter and height of seedlings growing in compressed

substrates

The stem diameters of cucumber seedlings receiving no fertilizer, irrigated with tap
water, and grown in all mixture formulations of compressed substrates had no
significant difference (P>0.05) at 20 days after germination (Figure 4(a)). Thus, the
 proportions of dairy manure vermicompost, smashed straw and peat could be optimized
to satisfy other parameters for seedling production in compressed substrates without
consideration for the stem diameter. However, the shoot lengths of cucumber seedlings
grown in compressed substrates had distinct differences between mixtures (Figure
4(b)). The shoot lengths of cucumber seedlings grown in eight formulations of
compressed substrate ranged from 15.83cm to 28.99cm, with the highest values found
in mixture TS2 (DMV: peat, 1:1 by volume). The shoot lengths of cucumber seedlings

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grown in compressed substrates, after incorporation of 0-50% smashed straw,
decreased by 14.6%. Upon addition of peat into DMV, the shoot lengths of cucumber

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seedlings increased with increasing the proportions of peat. However, too much rapid

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shoot growth can result in leggy, weak stems, susceptible to damage especially during
transplant (Korkmaz and Dufault 2001, Ming et al. 2011). This may be prevented by
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properly shrinking addition of peat into the DMV mixture (Figure 4(a)). In order to
control the shoot length and mitigate the effects of rapid growth in compressed
substrates, a mixture of the three materials should produce strong, healthy seedlings.
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Effects of the mixture on root and shoot dry weight of seedlings

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Figures 5(a) and 5(b) showed the contour and fitted plots for root and aerial dry
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weights of cucumber seedlings grown in compressed substrates, respectively. During

the experiment, we observed simultaneous profile changes in both root and
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aboveground dry weight, suggesting a positive correlation between these two metrics.
The zone of maximum response variables for root dry weight (Figure 5(a)) was located
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toward the side of triangle between the DMV and peat vertices, whereas the zone of
maximum response variables for aboveground dry weight (Figure 5(b)) was located
around the peat vertex. These results suggest that the addition of DMV and peat, but not
the addition of smashed straw, improves the dry weight response variables. However,
the inclusion of smashed straw was important for reduction of the breakage rate of
compressed substrates following seedling growth.
 Nutrition status of the cucumber seedlings grown in compressed substrates

The nutritional status of the cucumber seedlings is listed in Table 7. In cucumber

seedlings grown in different compressed substrates total nitrogen (TN), Mg and Cu
showed similar trends for each element between mixtures. A higher proportion of
smashed straw in the growth substrate negatively affected concentrations of these
elements in cucumber seedlings. In comparison, TN and Cu decreased dramatically,

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then rose again significantly in mixtures with adding peat into DMV, but it did not rise
above the levels found in seedlings grown in DMV-only substrate. Seedlings grown in

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compressed substrate containing all three materials had lower TN, Mg and Cu

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concentrations than seedlings obtained from substrate with DMV exclusively. The TK,
Ca and Mn contents of cucumber seedlings had no significant difference when the
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proportions of smashed straw, peat, or smashed straw with peat were increased. The
levels of TP distinctly increased commensurately with an increase in smashed straw or
peat, as well as when the three materials were mixed. The Zn content remained steady
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at range 57.56-60.24 mg kg-1 (no significance, P>0.05) and then decreased significantly
to 52.41mg kg-1 when straw was added from 0-50%, whereas the Zn concentration
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remained stable without significant change when 0-50% peat was added, though the Zn
concentration was lowest when all three components were included. Poole, Conover,
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and Joiner (1981) stated that the optimal ranges for potting plants with regard to TN, TP,
TK and Ca were 15-45, 1.5-3, 15-50 and 6-15 g kg-1, respectively. The TP and TK
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contents of cucumber seedlings in our study all fell within optimal ranges. The TN
values of TS1 and TS2 were within the optimal ranges and TS3-TS7 was close or below
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the optimal range. Ca values for TS1-TS4 were within the range while TS5, TS6 and
TS7 were slightly below. In general, Mn, Cu and Zn concentrations in cucumber
seedlings in our study, were far lower than the phytotoxic ranges given by Römheld and
Marschner (1991), and were within the ranges considered normal for plants (Adriano
2001).
 Optimisation of mixture proportion and Model validation

Increases in the proportions of DMV and peat, while reducing the use of smashed
straw helps to improve the response variables for shoot length, root dry weight and
aboveground dry weight, though the lack of this component will increase the BRA
response variable. Conversely, the smashed straw, when added in small amounts, will
decrease breakage variables (BRB and BRA) for compressed substrates. BRB and BRA

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were limited to 5%, in consideration of other factors such as shoot length, root dry
weight and aboveground dry weight while optimizing the proportions of DMV,

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smashed straw and peat. The expectations of shoot length, root dry weight and

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aboveground dry weight were the maximum observed values in the experiments,
respectively. A response optimizer was applied to generate the optimum proportion to
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meet the expectations of five responses. As the optimization plot (Figure 6) depicts,
the optimum ratio for these components was respectively 0.5917: 0.1608: 0.2475 when
the weight coefficients of the three parameters (Shoot length, Root dry weight and
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Aboveground dry weight) were 1: 1: 1. The composite desirability of the respective

mixtures was found to be 0.68036, which is mainly caused by the antagonistic effect for
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each response and the lower desirability of shoot lengths. However, the properly lower
shoot length of cucumber seedling makes it stronger which is more conducive to
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growth after transplanting.

In order to validate the model, experiments were conducted with the optimum
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compositions obtained. The results (BRB, BRA, shoot length, root dry weight and
aboveground dry weight) obtained from the experiments were found to be 0.72±0.13%,
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2.22±1.92%, 21.50±0.65cm, 95.52±0.85mg, 616.30±11.78mg, respectively. The

results obtained from the experiment were found to be comparable with the prediction.
The small variation in the results can be attributed to small change in the composition
of compressed substrates and the control of environment for seedlings used for the
validation experiments.
 CONCLUSIONS

From the results, it can be concluded that the application of the statistical mixture
designs for simultaneous compression effect of different compressed substrates and
production of root and shoot dry weight of cucumber seedlings grown in different
compressed substrates by using different agricultural wastes and peat as compressed

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substrates is an attractive process for improving the performance and to find the

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optimum formulations. Physical and chemical properties of compressed substrates (the

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bulk density, porosity, pH, EC, the elements content and so on), and the growth (shoot
lengths, stem diameter, shoot and root dry weights, and nutrition status) were evaluated

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in details as important factors on quality of compressed substrates for cucumber
seedlings production. As a whole, in this study with limitation of the simplex-centroid
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design, BRB and BRA of compressed substrates were both limited to 5% in the
production of compressed substrates, the optimum proportions of the three materials
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(dairy manure vermicompost, smashed straw and peat) could be determined at 0.5917:
0.1608: 0.2475 when the weight coefficients of the three parameters (Shoot length,
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Root dry weight and Aboveground dry weight) were 1: 1: 1. For different purpose, the
optimum ratio can be little changed on the base of different weight coefficient.
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ACKNOWLEDGMENTS
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The authors would like to acknowledge the financial supports of the Special Fund
for Agro-scientific Research in the Public Interest (201303091), and the Fundamental
Research Funds for the Central Universities (2662015QC002, 2015PY077,
2016PY018，2662016PY108).
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 ABOUT THE AUTHOURS
Hongliang Cao and Zhigang Liu are assistant professors, Qiaoxia Yuan is a
professor, Longyuan Yang and Shuai Luo are Ph.D. candidates in College of
Engineering, Huazhong Agricultural University, Wuhan, PR China.

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Figure 1. Constrained factor space inside a ternary diagram

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 Figure 2. The process of cucumber seedling production in different compressed
substrates: (a) the compressed substrates production; (b) the compressed substrates
after watering; (c) the 3th day after seeding; (d) the 20th day after seeding.

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 Figure 3. (a) The effects of different formulations on breakage rate before (BRB) the
seedling stage of different compressed substrates. (b) The effects of different
formulations on breakage rate after (BRA) the seedling stage of different compressed
substrates.

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 Figure 4. (a) The production of cucumber seedlings in compressed substrates. (b) The
effect of different formulations on shoot length.

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 Figure 5. (a) The effects of different formulations on root dry weight of cucumber. (b)
The effects of different formulations on aboveground dry weight of cucumber.

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 Figure 6. Optimization mixing proportion and composite desirability for responses of
BRB, BRA, shoot length (SL), root dry weight and aboveground dry weight based on
the optimal mixture proportions.

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 Table 1. Base physicochemical characteristics of the raw materials, (Means±SD),
n=3.
Particulars DMV Straw Peat
Wet basis moisture 25.12±2.62 17.23±1.62 14.78±1.12.
(%)
pH (1:5) 6.33± 0.03 6.44±0.45 5.99±0.33
EC(mS·cm-1) 2.04±0.02 0.14±0.05 0.22±0.01
TN (g·kg-1,dw) 9.51±0.62 1.21±0.45 2.90±0.10

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TP (g·kg-1,dw) 4.23±0.93 6.29±0.62 6.35±0.78
TK (g·kg-1,dw) 4.24±0.10 6.41±0.12 6.21±0.11

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Ca (g·kg-1,dw) 1.82±0.40 2.01±0.21 2.31±0.35

Mg (g·kg-1,dw) 1.80±0.07 1.16±0.05 2.44±0.04

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Mn(mg·kg-1,dw) 322.19±11.56 370.22±12.65 310.12±9.12
-1
Cu (mg·kg ,dw) 48.47±3.45 46.7±2.11 53.01±5.32
-1
Zn(mg·kg ,dw) 157.09±9.23
an 176.4±11.36 191.2±11.44
Note: EC Electrical conductivity, TN total nitrogen, TP total phosphorus, TK total
potassium, Ca calcium, Mg magnesium, Mn manganese, Cu copper, Zn zinc and dw
dry weight basis.
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 Table 2. Matrix of the simple centroid mixture design and response values for BRB,
BRA, shoot length, root dry weight and aboveground dry weight, (Means±SD), n=3.

Run Independent Independent variables, Y

variables, xij

DM SS Pea BRB BRA Shoot Root dry Aboveground

V t

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Restrictio [0.5, [0, [0, （%） （%） length(cm) weight dry weight

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n on 1] 0.5] 0.5] (mg) (mg)

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Factors x1 x2 x3

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TS1 1 0 0 47.68±10.32 18.06±9.82a
20.62±1.01b 61.70±2.12e 492.30±44.55d
a b
TS2 1/2 0 1/2
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2.71±0.05 c 20.83±5.89a 29.39±0.55a 91.90±1.69b 676.25±11.53a

TS3 1/2 1/2 0

29.03±0.06b 0±0c 20.33±2.37b 81.10±0cd 254.80±10.89e
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TS4 3/4 1/4 0

3.13±0.03c 0±0c 16.31±1.34c 72.00±3.96d 290.15±17.18e

TS5 3/4 0 1/4

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2.56±0.11c 8.33±3.92bc 21.92±2.02b 112.90±8.2a 610.15±8.98b

TS6 1/2 1/4 1/4 18.60±0.85b 89.05±6.01b 528.70±18.53c

1.52±0.17c 0±0c
c c d
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TS7 2/3 20.13±1.94b 90.15±1.48b

1/6 1/6 3.29±0.18c 0±0c 570.10±0.57bc
c c
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Note: TS test sample and SS smashed straw. BRB and BRA were the breakage
rate of compressed substrates before and after planting the cucumber seedlings,
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respectively.
 Table 3. Physical properties of compressed substrates, (Means±SD), n=3.
Formulation Bulk Water Air capacity Total Expansion
s density availability (% v/v) porosity (% rate
(g·cm-3) （% v/v） v/v)
TS1 0.49±0.03 a 47.24±9.33 b 6.49±2.92b 53.73±11.6 1.68±0.15 c
c 6b
TS2 0.32±0.01b 54.59±1.02 16.22±4.83 70.81±4.81 2.61±0.21 a
c ab a a

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TS3 0.41±0.04b 59.11±7.41a 5.05±1.52b 64.16±5.91 1.98±0.10b

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c b c ab c
TS4 0.45±0.04a 54.28±7.82 8.60±2.03b 62.8±6.60 1.76±0.13b

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TS5 0.40±0.07b 60.79±1.46 a 10.91±2.03 71.70±5.71 2.01±0.24 b

c ab a

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TS6 0.31±0.01 c 60.30±1.52 a 12.70±1.25 73.01±2.78 1.85±0.12b
ab a c
TS7 0.40±0.03b 59.29±1.33 3.12±2.14 c 62.41±2.48 1.89±0.15b
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c ab ab c

a, b, c,….: Means with the same letter in the same column are not
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significantly different (p>0.05).
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 Table 4. Chemical properties of different compressed substrates, means±SD, (n=3).
Compre pH EC TN TP TK Ca Mg Mn Cu Zn
ssed (1: (mS (g·k (g·k (g·k (g·k (g·kg , (mg·kg (mg·kg (mg·kg-1
-1 -1 -1

substrat 5) · g-1, g-1, g-1, g-1, dw) ,dw) ,dw) ,dw)

-1
es cm ) dw) dw) dw) dw)
TS1 6.33 2.04 9.52 4.23 4.24 1.82 1.80
322.19 48.47 157.09
±0.0 ±0.02 ±0.62 ±0.93 ±0.10 ±0.41 ±0.07 b
±19.20 b ±2.85 c ±5.88 b
3d a ab c b c

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TS2 6.50 1.23 6.67 6.20 4.02 2.54 1.94
290.33 49. 68 175.49

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±0.0 ±0.07 ±1.74 ±0.53 ±0.23 ±0.16 ±0.03a
±8.55 c ±1.87 bc ±18.20 ab
1a d b a b ab

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TS3 6.51 1.26 8.05 4.99 4.81 1.99 1.63
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350.26 47. 08 160.22

±0.0 ±0.13 ±1.19 ±0.41 ±0.29 ±0.08 ±0.05 c
±1.90 a ±0.95 c ±3.38 b
1a d ab bc a bc

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TS4 6.34 1.70 8.38 5.41 4.25 2.39 1.83
326.68 52.26 173.62
±0.0 ±0.05 ±2.11 ±0.20 ±0.13 ±0.23 ±0.08ab
±9.29 ab ±0.82ab ±5.29 ab
2d b ab ab b abc
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TS5 6.40 1.59 10.10 6.49 4.39 2.28 1.78
352.08 50.03 174.54
±0.0 ±0.13 ±1.76 ±0.54 ±0.09 ±0.55 ±0.12b
±21.54 a ±2.06 bc ±10.84 ab
5c bc a a ab bc
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TS6 6.44 1.26 8.59 5.67 4.47 2.99 1.87
328.68 51.84 174.37
±0.0 ±0.09 ±1.56 ±0.47 ±0.54 ±0.22 ±0.02ab
±15.28ab ±0.86 ab ±3.74 ab
2b d ab ab ab a
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TS7 6.40 1.48 8.75 6.00 4.49 2.46 1.93

346.12 53.27 183.85
±0.0 ±0.02 ±1.34 ±0.61 ±0.01 ±0.36 ±0.07 a
±14.26 ab ±1.56 a ±11.08 a
2c c ab ab ab ab
Note: Means followed by the different letter in the same column differed
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significantly according to Duncan’s multiple range test (p =0.05).

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 Table 5. Equations of the final reduced models for BRB, BRA, shoot length, root dry
weight and aboveground dry weight defined by the statistical mixture design.

Response Equations

Y  48x1  292x2  139x3  564x1 x2  362x1 x3  1191x2 x3 +1923x1 x2 x3

BRB(%)
(4)

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Y  18x1  52.4 x2  110.8x3  141.1x1 x2  174.5x1 x3  163.3x2 x3

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BRA(%)
(5)

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shoot Y  20.6x1  53.4x2  62.8x3  66.6x1 x2  49.3x1x3  390x2 x3 +579.7x1x2 x3

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length(cm) (6)

Root dry Y  62.1x1  111.2x2 -151.2x3  20.6x1 x2 +547.4x1 x3 +10.6x2 x3

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weight(mg) (7)

Abovegroun
Y  492x1  684x2  1653x3  1334x1 x2  1186x1 x3  8048x2 x3 +2064.75x1x2 x3
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d dry
(8)
weight(mg)
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 Table 6. Models, R2 and adjusted R2 and probability values for the final reduced
models (component proportions) of responses

Response Anova

Model F calculated F tabulated（0.05） R2 Adjusted R2 p-Value

BRB(%) Special cubic 43.71 4.39 97.40 95.17 <0.001

BRA(%) Quadratic 11.10 5.05 87.41 79.54 0.002

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shoot length(cm) Special cubic 13.53 4.39 92.06 85.26 0.002
Root dry weight(mg) Quadratic 30.33 5.05 94.99 91.86 <0.001

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Aboveground dry Special cubic 120.24 4.39 99.04 98.22 <0.001

weight(mg)

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 Table 7. Nutritional status of the cucumber seedlings grown in compressed substrates
Compres TN TP TK Ca Mg Mn Cu Zn
sed (g kg-1, (g kg-1, (g kg-1, (g kg-1, (g kg-1, (mg kg-1, (mg kg-1, (mg kg-1,
substrate dw) dw) dw) dw) dw) dw) dw) dw)
s
TS1 24.36± 1.05± 26.96± 6.51± 2.17± 18.88± 13.44± 57.56±
3.03 a 0.19 c 5.04 a 3.51 a 0.12 b 8.88 b 1.40 a 1.30 abc
TS2 19.23± 4.21± 23.97± 6.68± 2.54± 24.09± 11.73± 59.43±

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0.95b 0.45 a 4.83 ab 2.00 a 0.09 a 5.71 b 1.27 ab 4.74 ab

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TS3 12.19± 2.27± 22.97± 6.41± 1.99± 25.55± 8.18± 52.41±
0.10 c 0.60 b 1.56 ab 1.35 a 0.05 c 6.04 b 0.47 cd 2.18 c

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TS4 9.48± 2.84± 22.21± 5.19± 1.86± 36.95± 11.04± 60.24±

0.25 d 0.45 b 0.81 ab 0.91 a 0.05 cd 5.48 a 1.50 b 3.45 a

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TS5 12.15± 2.96± 20.51± 4.13± 1.89± 22.81± 10.09± 53.81±
1.09 c 0.85 b 4.97 ab 0.94 a 0.09 cd 2.55 b 0.43bc 5.77bc
TS6 9.80± 4.29± 19.70± 3.87± 1.60± 17.92± 6.87± 44.12±
0.27 d 0.38 a 2.06 b
an 0.84 a 0.08 e 3.23 b 1.07 d 1.35 d
TS7 9.21± 2.94± 21.52± 4.54± 1.78± 19.22± 7.78± 42.27±
0.06 d 0.71 b 3.64 ab 0.53 a 0.05 d 6.05 b 1.89 d 2.21 d
Normal 15-150 3-20 15-150
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range
Phytotoxi 400-2000 25-40 500-1500
c range
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Note: Phytotoxic range (Romheld and Marschner, 1991); normal range given by
Adriano (2001).
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