meat bone meal and fox manure as p sources for ryegrass (lolium multiflorum) grown on a limed soil

RESEARCH ARTICLE

Meat bone meal and fox manure as P sources for ryegrass(Lolium multiflorum) grown on a limed soil

Kari Ylivainio Æ Risto Uusitalo Æ Eila Turtola

Received: 1 August 2007 / Accepted: 5 December 2007 / Published online: 20 December 2007

� Springer Science+Business Media B.V. 2007

Abstract Phosphorus (P)-rich by-products, such as

meat and bone meal (MBM) and fur animal manures,

are potential P sources in plant production systems.

However, the solubility of P and its availability to

plants in these forms has not been evaluated. We

characterized P solubility in MBM, fox manures

(FoxM) and dairy manure (DairyM) by Hedley

fractionation and assessed P availability for ryegrass

in a pot experiment. Up to 81% of P was water-soluble

in DairyM, but only about 3 and 5–28% was soluble in

MBM and FoxM products, respectively. Of the P in

MBM and FoxM, 90 and 65–89%, respectively, was

soluble only in 1 M HCl. Most of the P was inorganic;

DairyM contained the highest share (14%) of organic

P. Based on ryegrass yields and P uptake in a 3-year

pot experiment with three P levels (25, 50 and

100 mg kg-1), P availability was equal in the DairyM

and superphosphate (SP) treatments. Compared with

the availability of P in DairyM and SP, 19 and 35–

54% of the P in MBM and FoxM, respectively, was

immediately available to the plant; for the 3-year

period with ten ryegrass cuts, the respective P

availabilities increased to 63 and 69–87%. Additions

of the sparingly soluble P sources MBM and FoxM

increased the acid-soluble P concentrations in the

experimental soil, with MBM having the strongest

effect. However, the acid-soluble P fraction decreased

with time. Although the immediate bioavailability of

P in sparingly soluble P sources was lower than that in

DairyM and SP, our results suggest that their use as a

long-term P supply for perennial plants in non-

calcareous soils should be encouraged.

Keywords Dairy manure � Fox manure �Meat and bone meal � P availability �P fractionation � Ryegrass

Introduction

As a result of bovine spongiform encephalopathy

(BSE), commonly known as ‘‘mad cow disease’’, the

EU banned the use of meat and bone meal (MBM) as

a feed for ruminants in 1994 and, subsequently, as a

feed for all livestock in 2001. In addition, EC

Regulation No. 1774/2002 of the European Parlia-

ment and the Council further restricted the use of

MBM as a fertilizer in 2002. Alternative methods for

handling MBM, such as incineration and cement

production, have been explored; however, in the

spring of 2006, restrictions on MBM usage as a

fertilizer were lifted in all regions of the EU.

The EU produces about 3 million tons of MBM

annually. MBM contains about 5% phosphorus (P),

mostly as calcium phosphate. However, due to its

chemical nature, the P in MBM is classified as being

K. Ylivainio (&) � R. Uusitalo � E. Turtola

MTT Agrifood Research Finland, 31600 Jokioinen,

Finland

e-mail: [email protected]

123

Nutr Cycl Agroecosyst (2008) 81:267–278

DOI 10.1007/s10705-007-9162-y

sparingly soluble. Another similar type of high-P

by-product in Finland is fur animal manure, which is

related to MBM because much of the P in fur animal

diets derives from MBM. Due to their high P

concentrations, both MBM and fur animal manure

are potential P sources for agricultural systems,

especially for organic farms specialized in plant

production. The annual manure P produced by

Finnish fur animal farms amounts to about 2 million

kilograms, whereas the amount of P associated with

MBM, eligible for fertilizer use, is about half of that.

If used solely as fertilizer, P derived from MBM and

fur animal manure could, respectively, contribute

about 0.5 and 1.0 kg of total P per cultivated ha in

Finland. In addition to these sparingly soluble P

sources, cattle manure comprises the most significant

source of manure-based P in Finland, with an average

annual input of 5.6 kg P per cultivated ha. These

numbers can be compared with P input, in the form of

mineral fertilizers, of 11–15 kg ha-1 (MMM 2004;

Antikainen et al. 2005).

In addition to P, MBM contains a considerable

amount of N (8%). The availability of this N to cereals

has been found to be close to that of mineral fertilizers

(Jeng et al. 2004), whereas the availability of P in

MBM is largely unexplored. In Norway, Jeng et al.

(2006) found that the relative efficiency of P in MBM

was about 50% of that of mineral P fertilizer for the

first crop of barley and ryegrass and that MBM had

residual P effects the following year. Fur animal

manure, in turn, has a long history as a P fertilizer in

Finland, and 40% of the P in fur animal manure is

considered to be available for plants. Hence, when fur

animal manure is used as a P source in plant

production, surplus application rates of total P are

common. In order to ensure environmentally and

economically sustainable methods of using these

sparingly soluble P sources in plant production

systems, the availability of P for plants in these

products needs to be further clarified. The necessity

for such studies is supported by the results of a recent

study showing that long-term surplus P applications,

as in the case of fur animal manure, had increased

soluble P fractions in soils down to a depth of 60 cm

(Uusitalo et al. 2007), strongly suggesting that P

derived from this source may be partly converted into

a water-soluble form and leach into lower soil profiles.

In this study, we examined the solubility of P in

MBM, fox manure processed to various extents

(FoxM) and dairy manure (DairyM). We also deter-

mined the effect of all three P sources on the growth

and P uptake of ryegrass in a P-deficient soil in a

3-year pot experiment. The availability of P in these

products was compared with a soluble P source,

superphosphate (SP). The aim of the study was to

provide a scientific foundation for the use of MBM,

FoxM and DairyM as P fertilizers. Ryegrass was

chosen as the test plant because of its efficient dry

matter production and P uptake (Brink et al. 2001).

Material and methods

FoxM was obtained from a commercial composting

facility (Natural Compost, Kaustinen, Finland) and

used as received or processed to produce composted

FoxM (cFoxM) or pelletized cFoxM (pcFoxM). To

produce cFoxM, FoxM was mixed with peat (1/1,

v/v) and placed in a batch composter in which air was

injected from the bottom. The mixture reached 70�C

within 24 h and was maintained at this temperature

for another 24 h, after which the compost was dried

with suction. During drying, the moisture content

decreased from 70 to 50%. The total duration of the

composting phase was 1 week. During pelletizing, the

cFoxM was dried with hot air (325�C) and com-

pressed through a 6-mm sieve, producing pellets

about 1 cm in length.

Honkajoki Oy (Honkajoki), the main facility for

handling animal carcasses in Finland, provided MBM

for the experiment. DairyM was taken from a manure

heap from MTTs barn (Agrifood Research Finland,

Jokioinen, Finland), where it had been composted

with peat and stored for a few months on a paved

plate.

Manure analyses

Total P concentrations of the manures and MBM

were analyzed with an inductively coupled plasma–

atomic emission spectrometer (ICP-AES; Thermo

Jarrel Ash, Franklin, MA) after microwave digestion

with aqua-reqia-HF (Lamothe et al. 1986). Total N

and C concentrations were analyzed with on a CN-

2000 analyzer (LECO, St. Joseph, MI).

The solubility of P was assessed using the Hedley

fractionation scheme as modified by Sharpley and

268 Nutr Cycl Agroecosyst (2008) 81:267–278

123

Moyer (2000). In brief, air-dried samples of manure

were finely ground using a mortar, and 1-g subsam-

ples were extracted, twice with water at a 1:60 (w/v)

ratio, then once with each of the following sequen-

tially: 0.5 M NaHCO3, 0.1 M NaOH and 1 M HCl.

Extraction times were 16 h, except for the first water

extraction, which was 4 h. Following extraction, the

samples were centrifuged (3000 g, 15 min), and

inorganic P (Pi) [supernatants filtered through a 0.2-

lm nucleopore membrane (Whatman, Maidstone,

UK)] and total P (unfiltered supernatant digested at

120�C with sulfuric acid and peroxodisulfate) con-

centrations in the supernatant were determined using

molybdate colorimetry (Murphy and Riley 1962).

The difference between total P and Pi in the

supernatant was assumed to represent organic P (Po).

Soil analyses

The soil used as growth medium was retrieved from

the plough layer of a Histic Podzol [as tentatively

classified according to the FAO (1998) system],

which had a low content of plant-available P,

according to Finnish agronomic criteria (Table 1).

Soil texture was analyzed with a pipette method, as

described by Elonen (1971), C concentration was

analyzed with the LECO CN-2000 analyzer, and soil

pH was determined in a 1:2.5 (v/v) water suspension.

Plant available P, Ca, K and Mg concentrations were

analyzed according to the Finnish agronomic soil

testing protocol [acid ammonium acetate (AAAc);

Vuorinen and Makitie 1955], and P content was also

determined with 0.5 M NaHCO3 (Olsen-P). In addi-

tion, soil P was analyzed with a modified Hedley

fractionation scheme as described above.

Growth experiment

Ryegrass (Lolium multiflorum var. Turgo) was grown

under a glass roof outdoors at ambient air tempera-

ture on sandy soil for 3 consecutive years. Before

establishing the experiment, the soil was air-dried and

passed through a 6-mm sieve to remove coarse

fragments and root debris. The soil (6.5 kg) was then

limed with an amount of CaCO3 (6.85 g) that was

calculated to bring the pH to 6.5.

Phosphorus sources are listed in Table 2. Phos-

phorus application rates were 25, 50 and 100 mg kg-1

soil, corresponding to 162.5, 325 and 650 mg pot-1,

respectively. The P sources were applied only once, at

the beginning of the experiment, and mixed into the

whole soil volume. All the P sources, except SP and

MBM, were air dried and passed through a 6-mm

sieve before application. The control treatment did not

receive P. Treatments were replicated four times.

In the first year, all of the pots received the

following amounts of nutrients during sowing (mg

pot-1): 1000 N (as NH4NO3 and KNO3), 1000 K

(KNO3), 200 Mg (MgSO4), 20 Na (NaCl), 10 Fe

(FeSO4), 10 Zn (ZnSO4), 10 Mn (MnSO4), 5 Cu

(CuSO4), 1 B (H3BO3) and 1 Mo (Na2MoO4). In the

following years, the same amounts of nutrients were

applied during the sowing, with the exception that the

amounts of applied N and K were 1500 mg pot-1.Table 1 Characteristics of the soil used in the pot experiment

Soil texture (%)

0.2–2 mm 18

0.02–0.2 mm 67

0.002–0.02 mm 10

\0.002 mm 5

Total C (%) 2.1

Soil pH (water, 1:2.5, w/v) 5.9

Olsen P (mg kg-1 soil) 18.6

AAAc extractable nutrientsa (mg l-1)

P 3.2

Ca 815

K 169

Mg 57

a Acid ammonium acetate, pH 4.65

Table 2 Total phosphorus (P), nitrogen (N), calcium (Ca) and

carbon (C) concentrations and dry weight (DW) percentage

content of the air-dried amendments used in the pot experiment

P sourcea P

(mg g-1)

N

(mg g-1)

Ca

(mg g-1)

C

(mg g-1)

DW

(%)

MBM 62.7 71.2 141.5 338.0 96.5

FoxM 34.3 34.1 60.1 276.4 91.2

cFoxM 25.7 34.1 48.9 321.9 92.1

pcFoxM 24.1 31.6 44.2 278.4 91.0

DairyM 3.8 20.5 8.8 382.6 89.7

SP 90.8 – 200.9 7.1 97.9

a MBM, Meat and bone meal;; FoxM, fox manure; cFoxM,

composted FoxM; cpFoxM, pelletized cFoxM; DairyM, dairy

manure; SP, superphosphate (soluble P source)

Nutr Cycl Agroecosyst (2008) 81:267–278 269

123

Ryegrass was sown in the same pots three

consecutive springs, with a total of about 50 germi-

nating seeds per pot, and irrigated with deionized

water. The ryegrass was cut four times in the first

2 years, but only twice in the third year, due to poor

growth. Each year the final cuttings were carried out

after growth had ceased due to low temperature. Each

ryegrass stand was cut 2 cm above the soil surface,

washed briefly with detergent (0.1% Deconex), rinsed

three times with deionized water, blotted dry and

dried at 65�C in a forced-draught oven and ground

with a hammer mill. The P concentration in plant

material was analyzed after HNO3 (about 7 M)

digestion, with ICP-AES, and N was analyzed using

the Kjeldall method. In-house reference material was

included in the analyses.

After each cut during the growth period, supple-

mental doses of N, K and Mg were applied to all pots

to ensure their sufficiency for the following crop. In

the first year, the supplements per pot included

800 mg of N and K and 100 mg of Mg. In the second

year, 1500 mg of N and K and 100 mg of Mg were

applied per pot after the two first cuts, and 500 mg of

N and K and 50 mg of Mg was added after the third

cut. In the third year, 1000 mg of N, 500 mg of K and

50 mg of Mg were applied after the first cut. For the

second and the third year, the supplemental doses of

K and Mg were determined according to the AAAc

extractable concentrations of K and Mg in soil

samples taken from the pots at the end of the

previous years (after the fourth and eight ryegrass

cuts; data not shown). Additional liming was done

before the seeding in the third year due to a decline in

soil pH. Soils with a pH lower than 5.6 (the lowest

value was 5.1, obtained for the control treatment)

received 10 g of CaCO3, while the other soils

received 7 g. After the experiment was terminated,

soil pH varied from 6.0 (control) to 6.5 (cFoxM,

50 mg P kg-1).

After the last cut in each year, soil sample cores

were taken from each pot through the entire soil

column (about 0.5 l pot-1). The soil in each pot was

then sliced horizontally through the middle and the

top half turned upside down on top of the bottom half.

The soils were stored over winter at ambient air

temperature, covered with plastic under a glass roof.

In the spring, the soils were passed through a 6-mm

sieve and the ryegrass roots were placed at the bottom

of the pots.

To estimate P availabilities from different P

sources, data on ryegrass yields and P uptake for

the SP and DairyM treatments were used to solve the

following equation: y = A + B*(1-e- Cx), where A is

the minimum ryegrass yield or P uptake, B is the

maximum yield or P uptake response, C is a

coefficient and x the amount of applied P (mg kg-1

soil). This equation was then used with the data for

sparingly soluble P treatments to calculate the

amount of SP- or DairyM-based P required to

produce the corresponding yield or P uptake. These

values were then made proportional to the amount of

P applied.

Results and discussion

Phosphorus content and solubility of P sources

Of the by-products studied here, MBM had the

highest and DairyM the lowest total P concentration

(Table 2). The composting of FoxM decreased the

total P concentration due to the addition of peat prior

to composting, while pelletizing the compost had no

further effect on total P concentration (Table 2).

Phosphorus solubility varied widely between the

different P sources (Table 3). In DairyM, 81% of the

sum of the P fractions was water-soluble compared to

87% in SP; in contrast, water-soluble P was a minor

fraction (3–28%) in MBM and fox manure products,

with 65–90% of the P being in acid-soluble form

only. A high share of acid-soluble P in the FoxM is in

line with the diet of foxes, which at most may derive

30% of its P content from MBM; Baltic herring as

well as other dietary sources also contain P of low

solubility.

Composting of FoxM had no influence on P

solubility, but pelletizing cFoxM decreased both the

content of water-soluble P (Table 3) and total N

concentration (Table 2). During pelletizing, the com-

post was dried with hot air (325�C), which may alter

P solubility. The results are in agreement with the

study of O’Connor et al. (2004), who found that the

pelletizing of biosolids decreased water-soluble P

fractions and increased NH3 volatilization.

Most of the P in the studied by-products was

inorganic. Only in DairyM was a notable proportion

(14%) of P present as Po (Table 3), and most of it

was water- and NaHCO3-extractable, summing to


123

about 12% of the total P fractions. This results agrees

with that reported by Sharpley and Moyer (2000) who

found Po to be 14% of the total P extracted in water-

and NaHCO3-fractions from dairy manure.

Differences in P solubility in manures are related

to the diet and age of the animals, the bedding

material and the handling and storage of the manure.

However, the composting of FoxM had no influence

on P solubility, indicating that bone-derived P was

not affected by composting. Similar results were

obtained with poultry manure by Dao et al. (2001),

who argued that the solubility of dicalcium phos-

phate, added as dietary mineral supplements, is not

affected by composting. The DairyM in the present

study was mixed with peat and stored on a paved

plate for a few months. While the pile was rather

dense, the composting was probably minimal and

hence the manure can be regarded as stockpiled

manure.

Ryegrass yields in the pot experiment

All P sources increased the total sum of the ten

ryegrass yields significantly compared to the control

treatment (Table 4) with MBM having the least effect

(25 mg P kg soil-1, 27%) and DairyM have the

largest effect (100 mg P kg soil-1, 120%). Both

cFoxM and FoxM increased ryegrass yields equally,

and pcFoxM gave significantly lower yields than

FoxM only in the first and fourth cuts. Therefore, we

present yield data only for cFoxM.

Among the invidual cuttings, the most significant

differences in ryegrass yields were obtained in the

first cut, where DairyM gave the highest yields in all

P application levels, followed by SP [ cFoxM [MBM (Table 4). Thereafter, yield differences leveled

off. The lower yields of the SP treatment compared to

DairyM treatment were partly related to N availabil-

ity. All treatments received the same base level of

inorganic N, but during the first year it may have been

too low for maximum ryegrass growth, especially at

the highest P level (SP treatment), whereas organic

amendments provided N as well. This is supported by

the fact that among all the treatments in the first cut,

the lowest level of N was found in the 100 mg SP–

P kg-1 (19.1 mg N g-1 DW) treatment, which was

significantly lower than that in the 25 mg SP–P kg-1

(25.2 mg N g-1 DW) treatment. The same phenom-

enon was evident with other P sources as well, but

was least pronounced with DairyM (24.6 and

24.5 mg N g-1 DW in 25 and 100 mg DairyM–

P kg-1 treatments, respectively). Depressed N avail-

ability in SP treatments resulted in equal total N

uptakes for the 25 and 100 mg SP–P kg-1 treatments

(739 vs. 747 mg pot-1, respectively), whereas the P

uptake almost doubled (Table 5).

In the second experimental year, there was an

overall decline in the yields, which decreased more in

the two lower P application levels (25 and 50 mg P

kg-1 soil) than in the 100 mg P kg-1 treatments. At

the highest P application level, the yield depression

was most evident in the third year, indicating a

decrease in plant available P. This was supported by

the prolonged time to reach a given growth stage.

Most of the differences between the ryegrass

yields in the first 2 years were observed in the first

cuts of the growing seasons (Table 4, first and fifth

cuts). For example, with the highest P application

levels of MBM and DairyM, yield differences in cuts

Table 3 Inorganic (-i) and organic (-o) P concentrations in air-dried P sources according to the Hedley fractionation scheme

MBM (mg g-1) FoxM (mg g-1) cFoxM (mg g-1) pcFoxM (mg g-1) DairyM (mg g-1) SP (mg g-1)

Pw-i 1.0 6.9 5.6 0.5 3.2 84.8

Pw-o 0.8 1.5 1.4 0.7 0.3 –

PNaHCO3-i 2.3 1.7 0.8 0.7 0.2 0.6

PNaHCO3-o 0.1 0.1 0.3 0.2 0.2 –

PNaOH-i 0.4 0.3 0.2 0.4 0.1 5.3

PNaOH-o – 0.3 0.3 0.2 0.1 –

PHCl-i 51.3 19.2 17.5 22.6 0.2 7.1

RP-i 55.9 28.0 24.2 24.2 3.7 97.8

RP-o 0.9 1.7 1.9 1.1 0.6 –


123

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123

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38

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5.5

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61

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31

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5.8

a

MB

M

25

46

.8ab

46

.5ab

33

.7ab

c4

1.2

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16

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7.5

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8.9

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c6

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f5

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f2

9.5

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ryM

25

48

.9ab

c6

4.7

def

36

.5b

cd4

1.5

abc

19

1.6

cd1

7.9

a3

0.4

ab2

2.8

ab1

5.8

b8

7.0

ab1

7.9

ab1

9.5

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7.4

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16

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50

65

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.0fg

47

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51

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e2

35

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123

number one and five accounted for 65% of the total

yield difference during the first two years. Compared

with the SP and DairyM treatments, however, cFoxM

and MBM treatments increased ryegrass yields as the

growing season progressed, especially in the second

year (Table 4).

The importance of P availability at the onset of

growth was evident when comparing the sparingly

soluble P sources with SP and DairyM in the first

ryegrass cuts of the first and second growing seasons.

At the lowest P application level, DairyM increased

ryegrass yields the most in the first cut (Table 4),

even though total P uptake was at the same level as

for the other P sources (Table 5). For example,

treatment with DairyM produced a 55% higher

ryegrass yield than treatment with MBM with almost

the same total P uptake. Total P uptake increased in

DairyM and SP treatments with increased P applica-

tion level, whereas it remained about the same at the

two lowest P application levels for the cFoxM and

MBM treatments (Table 5). However, ryegrass yields

increased significantly as the P application level of

MBM and cFoxM increased from 25 to 50 mg kg-1 soil

(Table 4). This suggests that as the P application level

increased, ryegrass P needs were better met at the

beginning of the growth period and the P enhanced dry

matter production. It has been suggested that early P

acquisition is critical for producing optimal yields, while

later application of P does not improve growth (Grant

et al. 2001). However, from the second cut onwards,

sparingly soluble P sources clearly increased both the P

uptake and the ryegrass yield. This result indicates that

the subsequent growth potential of ryegrass was not

depressed by the lower availability of P in the MBM and

cFoxM during the initial growth stage, as evidenced by

the increased ryegrass yields in the later cuts. Similar

results were obtained by Baker et al. (1989) with bone

meal, who observed that corn yield and P uptake

increased in consecutive growth periods and that from

the second harvest onwards, yield and P uptake were

equal to those obtained with Ca(H2PO4)2.

Ryegrass has a high root–shoot ratio and is

referred to as a P-efficient species with relatively

low external P requirements (Fohse et al. 1988).

Phosphorus uptake by ryegrass has been shown to

increase if the availability of P increases after a

period of deprivation, probably due to a deprivation-

induced increase in the number of P uptake sites in

the roots (Breeze et al. 1985). In addition, ryegrass

growth is enhanced by the presence of apatite in acid

soil (Junge and Werner 1989), and Chien and Menon

(1995) suggested that apatite is a suitable P source for

plants with a long growth period and a low external P

requirement. Therefore, MBM and the processed fox

manures may be convenient storage P fertilizers for

plants with the above characteristics. This would

reduce the need for surface application of P, especially

with perennial crops, and ultimately abate the problem

of P transport by runoff (Turtola and Yli-Halla 1999).

Availability of P in the different P sources

When DairyM was mixed in soil, P availability was

equal to that of SP, whether it was estimated

according to ryegrass yields or by P uptake. This is

in line with the high proportion of water-soluble P in

DairyM. In addition, manure may depress soil P

sorption strength (Holford et al. 1997) as manure-

based organic acids may compete for the same

binding sites as P (Øgaard 1996; Haynes and

Mokolobate 2001). This may have been the reason

that the highest water-extractable P concentration

was obtained for the 100 mg P kg-1 DairyM treat-

ment at the end of the experiment (Table 6), despite

the fact that this treatment gave the highest level of P

uptake. DairyM and SP can be both considered as

reference P sources when evaluating the P availabil-

ity in the sparingly soluble P sources, because they

had the same level of P availability. In addition, N

and C effects on P acquisition are within the range of

that in SP and DairyM. The application levels of P

correlated well with ryegrass yields in the SP and

DairyM treatments for the 3-year period (R2 [ 0.99).

Phosphorus availability from sparingly soluble

sources, when judged by yields, increased as the

experiment proceeded. According to the data from

the very first ryegrass cut, P availability was 19, 35,

43 and 54% in the MBM, pcFoxM, cFoxM and FoxM

treatments, respectively, where the availability of P in

the sparingly soluble P sources was taken as an

average of the three P application levels. However, in

the fourth cut, the respective values were 65, 50, 71

and 100%. For the sum of the ten ryegrass cuts, the

respective P availabilities were 63, 69, 74 and 87%.

When judging P availability based on total P uptake,

higher values were obtained (data not shown). This is

probably related to the low acquisition of P during the


123

early development of the ryegrass, which depressed

ryegrass growth.

Enhancement of ryegrass growth due to N derived

from the P amendments was most evident in the first

ryegrass cut. At the highest P application level, N

uptake in the DairyM treatment was 420 mg more

(total N uptake 1168 mg pot-1) than in the SP

treatment (total N uptake 747 mg pot-1). This prob-

ably stimulated both growth and P uptake, as shown by

Belanger et al. (2002). For the other organic amend-

ments, N derived from MBM was mostly in a plant-

available form (C:N ratio 4.7:1), as evidenced by the

approximately 250 mg greater N uptake from MBM

(100 mg P kg-1) than from SP (100 mg P kg-1) dur-

ing the first year. This difference in N uptake in the first

year accounted for 33% of the total N content in the

applied MBM. Moreover, the N uptake for MBM was

the second highest (after the DairyM treatment) in the

first ryegrass cut, even though MBM increased

ryegrass yield the least. This result is in line with

observations that N in MBM was equivalent to at least

80% of the N in mineral fertilizer (Jeng et al. 2004).

In the three succeeding years, P concentration in

the ryegrass stand varied between 1.46–2.69, 1.01–

1.88 and 0.90–1.12 mg g-1 dry weight (DW), respec-

tively, excluding the eighth and tenth cuts (data not

shown, values can be calculated by dividing the

values in Table 5 with the values in Table 4). Due to

the cessation of growth caused by low temperature,

the eighth and tenth ryegrass cuts were done at an

earlier growth stage, and thus P concentrations were

higher, varying between 1.87–3.18 and 1.21–1.57 mg

g-1 DW, respectively. Phosphorus concentrations of

ryegrass did not correlate with the respective yields

during the experiment (R2 = 0.03), indicating that P

concentration was a poor indicator of the physiolog-

ically available P content, due to the dilution effect

caused by enhanced growth.

In this study, ryegrass growth became depressed

before P concentration decreased below 1 mg g-1

DW, which is the concentration believed to indicate a

severe P deficiency (Yli-Halla 1991). The N: P ratio

of the crop may be a better indicator of the P status of

plants than P concentration. According to Gusewell

(2004), biomass production is depressed by P short-

age when the N: P ratio is[20. In this study, the N: P

ratio was up to 41 in the ninth cut (data not shown),

indicating a severe P deprivation.

Solubility of phosphorus in the experimental soil

After ten ryegrass cuts, those P fractions showing the

greatest decrease were the water- and bicarbonate-

Table 6 Hedley fractionation scheme for experimental soils after first year (four cuts) and third year (ten cuts) in 100 mg P kg-1

treatments

Pwater PNaHCO3 PNaOH PHCl RPi and Po

Pi Po Pi Po Pi Po Pi

Beginning of the exp. 4.4 7.8 37.1 30.0 105.9 112.5 176.5 474.2

Treatment

After 1st year

Control 2.3 a 7.7 ab 29.1 abc 42.7 d 96.7 b 108.8 a 173.1 ab 460.4 ab

MBM 5.4 bc 12.6 c 38.3 de 31.3 ab 103.0 bc 125.3 b 245.0 d 560.9 ef

cFoxM 5.7 c 12.4 c 42.4 ef 38.6 cd 118.0 de 140.7 cd 215.3 cd 572.9 f

DairyM 6.0 c 19.1 d 37.9 cde 32.8 abc 111.1 cd 122.0 ab 173.6 ab 502.5 bcd

SP 6.4 c 10.2 bc 47.6 f 38.0 bcd 129.8 e 127.9 bc 175.2 ab 535.2 cdef

After 3rd year

Control 0.9 a 6.4 a 21.8 a 29.6 a 79.0 a 121.1 ab 166.5 ab 425.3 a

MBM 2.4 a 10.5 bc 26.2 ab 28.8 a 92.7 ab 127.0 bc 228.0 cd 515.6 bcdef

cfoxM 2.3 a 10.5 bc 25.1 ab 32.6 abc 112.7 cd 119.8 ab 207.2 bc 510.2 bcde

DairyM 2.9 ab 12.7 c 25.1 ab 32.7 abc 89.2 ab 145.9 d 173.7 ab 482.3 abc

SP 2.0 a 11.7 c 30.7 bcd 34.8 abc 121.0 de 133.9 bcd 214.9 cd 549.1 def

Columns with the same letter do not differ significantly (P [ 0.05, Tukey’s test)


123

extractable Pi fractions – 80 and 41%, respectively –

in the control treatment (no P added) (Table 6).

However, these two fractions represented only 8% of

the sum of the Pi fractions. Quantitatively, the NaOH-

extractable Pi fraction decreased the most, by

26.9 mg kg-1 (25% decrease). Only the acid-soluble

P fraction was unaltered in the control treatment. As

for the P-amended soils, the highest P application

level (100 mg P kg-1) affected the concentrations of

the different P fractions most and, therefore, only the

results of 100 mg P kg-1 treatments are presented

(Table 6). In addition, the P solubility in soils after

the addition of the different fox manures (FoxM,

cFoxM, pcFoxM) was rather similar, and only the

results for cFoxM are presented.

After the first growth period, the water- and

NaHCO3-extractable Pi concentrations in the soils

were significantly higher due to P applications, but

thereafter the influence of the treatments on these

fractions was less evident. Throughout the treatment

period, the NaHCO3- and NaOH- extractable Pi

concentrations remained significantly higher only in

the SP treatments, and the NaOH-extractable Pi

concentrations remained significantly higher only in

the cFoxM treatment (Table 6). The acid-soluble Pi

fraction increased the most in the MBM and cFoxM

treatment. This is in line with the high share of acid-

soluble P in these P sources. During the two first

years, DairyM and SP had no influence on the acid-

soluble Pi fraction. During the last year, however, this

fraction increased somewhat in the MBM, cFoxM

and SP treatments. This may be due to the liming

with CaCO3 before the last seeding. A similar

increase in the acid-soluble P fractions after liming

was detected by Guo et al. (2000), who also found

that the Ca–P formed was not stable in highly

weathered soil.

Phosphorus in MBM is mostly in the form of

calcium phosphate, and its solubility is partly

governed by soil pH. After the first growth period,

about 70% of the added P in the MBM treatment

(100 mg P kg-1) was still in acid-soluble form. In

the succeeding years, the amount of this P fraction

decreased, but after 3 years about 60% of the added P

still remained in acid-soluble form. This result shows

that in our non-calcareous soils, MBM-derived P

gradually converted to a more soluble form. A recent

study has shown that a long history of fur animal

manure application on non-calcareous soils increased

soluble P fractions up to a depth of 60 cm (Uusitalo

et al 2007), suggesting that P derived from fur animal

manure gradually becomes water-soluble and leaches

down into the soil profile.

Of the P amendments used, DairyM increased

water-extractable Po concentrations of the soil the

most. After the first growth period, the water-

extractable Po concentrations in the DairyM treat-

ments increased from an initial value of 7.8 mg kg-1

to final values of 11.6 mg kg-1 (25 mg P kg-1) and

19.1 mg kg-1 (100 mg P kg-1). These values were

significantly higher than that found in the control

treatment (7.7 mg kg-1). The increased concentra-

tion of water-soluble Po with the DairyM treatment

(100 mg P kg-1) corresponded to the amount of

water + bicarbonate-extractable Po contents in the

added DairyM. In the succeeding years, the content of

water-soluble Po in the experimental soil declined,

indicating mineralization.

Variations in the NaHCO3- and NaOH-extractable

Po concentrations during the experiment were prob-

ably related to both the characteristics of the P

amendments and the amount of organic acids pro-

duced by ryegrass roots due to P deficiency and their

influence on microbial activity in the soil. While the

amount of C applied was the highest in the DairyM

treatment (Table 2), the mineralization of organic

matter may explain the steady increase in NaOH

extractable Po to its maximal level at the end of the

experiment in DairyM treatment. Other possible C

sources for soil microorganisms are organic acids,

which are excreted by roots after a period of P

deficiency (Jones 1998) and may increase the growth

of microorganisms and enhance microbial activity in

the rhizosphere (Toal et al. 2000). Increased micro-

bial activity in the rhizosphere may also increase Po in

the NaHCO3- (Helal and Sauerbeck 1984; Chen et al.

2002) and NaOH-extractable fractions (Zoysa et al.

1997, 1999). According to Guo et al. (2000),

NaHCO3–Po starts to contribute to the plant-available

P pool after the soil-available Pi pool is exhausted. In

the control treatment, there were indications of these

events, with an increase of NaHCO3–Po from 30 mg

kg-1 to 42.7 mg kg-1 during the first growth period,

and a subsequent decline to 32.1 mg kg-1 after the

second growth period.

Sparingly soluble P sources maintained the AAAc-

extractable P concentration at elevated levels

throughout the experiment (Table 7). However, lower


123

ryegrass yields in the MBM and cFoxM treatments

compared to the DairyM and SP treatments demon-

strate that the concentrations of AAAc-extractable P

were not in agreement with the plant availability of

P in soils supplemented with these sparingly soluble

P sources. The water-extractable P concentrations

were also lower in both sparingly soluble P sources

(Table 3) and in soils amended with these products,

although the difference was not significant when

compared with the respective values of the DairyM

treatment (Table 6). Overestimation of the concen-

tration of readily soluble P by the AAAc extraction in

soils amended with these sparingly soluble P sources

was probably due to the dissolution of calcium

phosphates by AAAc (pH 4.65). In the study of Jeng

et al. (2006), the amount of readily available P in

MBM was estimated to be 33–40% of the total P

when the estimation was based on the extraction of

MBM with ammonium lactate (pH 3.75). These

results suggest that extracting calcium phosphate

compounds, or soils amended with these compounds,

does not reflect their immediate P availability for

plants but rather gives an idea of their long-term

availability. In the long term, the concentrations of

water-extractable P may also increase in soils

amended with sparingly soluble P sources when the

amount of P added continuously exceeds the crop P

uptake (Uusitalo et al. 2007).

Conclusion

This study showed that MBM and fox manure may

represent good P storage fertilizers for plants which

have a long growing period and efficient P uptake

systems. The application of such fertilizers would

reduce the need for surface application of P for

perennial grasses, subsequently reducing P runoff.

Moreover, MBM and fox manure could be used

instead of inorganic fertilizers to improve the recy-

cling of P, and in organic farms lacking animal

production and their own manure source. Although

most P in MBM and fox manure was initially in an

acid-soluble form, P eventually converted to a plant-

available form and sustained ryegrass yields compa-

rable to those achieved with DairyM or SP. However,

plant growth as well as nutrient uptake is more

intense in pot experiments than in the field. There-

fore, field studies are underway to further evaluate the

P availability from these sparingly soluble P sources.

Acknowledgements We thank research assistant Pirkko

Maki and laboratory technician Anja Lehtonen for their

skillful technical assistance with the pot experiment and the

related analyses. The Ministry of Agriculture and Forestry,

the Finnish Fur Breeders’ Association and Honkajoki Oy are

gratefully acknowledged for funding.

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P source AAAc

After first year After third year

Control 2.3 ab 1.7 a

MBM 9.6 e 4.8 cd

cFoxM 7.8 e 4.2 bcd

DairyM 5.1 d 2.6 ab

SP 5.4 d 2.9 abc

Averages with the same letter do not differ significantly

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meat bone meal and fox manure as p sources for ryegrass (lolium multiflorum) grown on a limed soil

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