AMINO ACID PROFILE OF EARTHWORM AND EARTHWORM MEAL

(Lumbricus rubellus) FOR ANIMAL FEEDSTUFF

L. Istiqomah, A. Sofyan, E. Damayanti and H. Julendra

Division of Feed and Animal Nutrition
Research Unit for Development of Chemical Engineering Processes
Indonesian Institute of Sciences (LIPI)
Jl. Jogja - Wonosari Km. 31, Gading, Playen, Gunungkidul, Yogyakarta, 55861
Corresponding Email: ps_uty@yahoo.com

Received November 3, 2009; Accepted November 30, 2009

ABSTRACT

Earthworm meal (Lumbricus rubellus) has become one of natural material that could be used as
feed additive. Powdering method of earthworm was done by using formic acid addition. The study was
carried out (1) to evaluate the essential amino acid profile of earthworm and earthworm meal, (2) to
calculate the value of essential amino acid index (EAAI) of both materials. A modified EAAI equation
was developed from the essential amino acid profile of earthworm and earthworm meal. The result
showed that essential amino acid of earthworm was dominated by histidine (0.63% of dry matter basis),
meanwhile the earthworm meal was dominated by isoleucine (1.98% of dry matter basis). The non
essential amino acid of earthworm and earthworm meal was dominated by glutamic acid (1.52% and
3.60% of dry matter basis respectively). The value of essential amino acid index obtained from
earthworm meal was higher (58.67%) than those from earthworm (21.23%). It is concluded that
powdering method of earthworm by using formic acid addition had higher amino acid balance than
earthworm.
Keywords: amino acid, earthworm meal, feed additive L. rubellus

INTRODUCTION The biological value has positive correlation

with amino acid balance (Rama Rao et al., 1964).
Fish meal is the conventional source of Meanwhile the amino acid balance could be
animal protein in poultry feed and it has been calculated by Essential Amino Acid Index (EAAI)
valued for its balanced amino acids, vitamin (Oser, 1951 cit. Mitchell and Block, 1946). The
content, palatability and growth factors (Tacon, increase of EAAI value will enhance nutrient
1993). Because of the increasing cost of high absorption (Raghunath and Narasinga Rao, 1984).
quality fish meal required for poultry feed and due There were some treatments that could increase
to declining stocks of fish from capture fishery the EAAI value in physically or mechanically
and competition for feed in animal husbandry, way, one of that was powdering method.
there is now need to search for alternative sources Previous study showed that earthworm meal
of animal protein for poultry feeds especially in of Lumbricus rubellus had 65.63% crude protein
developing countries (Sales and Janssens, 2006). content (Damayanti et al., 2008), earthworm meal
These alternatives must be able to supply of Lumbricus terestris contained 32.60% protein
adequate indispensable and dispensable amino (Julendra, 2003), and earthworm meal of
acid requirements of the fish or sufficient amino Perionyx excavatus contained 57.2% crude
nitrogen to enable their synthesis. protein and had complete amino acid (Tram et al.,
Many scientists have reported the possible 2005) Although the special quality of earthworm
use of some alternative animal protein feedstuffs meal has been known well as potential fish meal
to fish meal such as earthworm meal (Hilton, replacement as protein source (Yaqub, 1991), but
1983; Yaqub, 1991; Edwards, 1998; Sogbesan and it’s still necessary to study about the essential
Ugwumba, 2006; Sogbesan and Madu, 2008). As amino acid composition in earthworm meal.
a feed supplement, it has been found that The experiments were carried out (1) to
earthworm is equal to or surpass fish meal and evaluate the profile of essential amino acid
meat meal as an animal protein source for poultry. composition of earthworm and earthworm meals,

Amino acid of earthworm meal ( L. Istiqomah et al.) 253

(2) to the value of essential amino acid index
Table 1. Nutrient Composition of
(EAAI) of both materials. Earthworm Meal
MATERIALS AND METHODS
Composition Earthworm meal
Moisture 9.03
Earthworm Meal Preparation
Based on United States patent Crude Protein 63.06
US005128148A (Ishii et al., 1992), disclosed Ether Extract 18.5
processing for the production of dried earthworm Crude Fiber 0.19
powder which comprises the steps of leaving a Nitrogen Free Extract 12.41
species of 1 kg earthworms Lumbricus rubellus, Organic Matter 94.16
Allolobophora caliginosa or Pheretima Ash 5.81
comunissima in 2 – 4 litres fresh water or slightly
acidic aqueous solution such as malate acid, * Dry matter basis (%)
citrate acid, phospate acid, tartate acid, lactic acid
and succinate acid with comparison of pH average
5.2 – 6.2, kept in 8 – 150C temperature for 2.5 – neutralized by NaOH 6N addition. The sample
18 hours until the alimentary canal thereof is freed was filtered by 0.45 μm millex filter paper using
of soil, wet-grinding the earthworm, and freeze- vacuum filter. Before the sample was injected to
drying and then vacuum drying the resulting the HPLC, 275 μL OPA solution (0,01 g
suspension under a vacuum of 10 mmHg or phetaldialdehyde, 9 ml methanol, 40 ml buffer
below for 10 to 100 hours while raising the borax pH 9.1, 100 μL 2-mercaptoethanol were
temperature gradually from -600C to 800C. This added into 25 μL sample, vortex and being
steps would produced dried earthworm powder. reacted for 5 minutes. The HPLC operation was
Based on Edwards (1985) method, the C18 column, move phase A: Sodium acetate pH
earthworm washed using water to separate 5.0; B : 80 MeOH / 15 Buffer acetate / 5 THF
manure from outside skin and fecal (fecal mud) (v/v/v), rate flow 1.5 ml/minute, detector :
then kept in cold water (140 C) for 24 hours. fluorescence λ excitation 340 nm; λ emission 450
Formic acid (80%) was added around 3% of nm.
earthworm body weight. The earthworm was
grinded and dried using oven 500C for 12 hours Observed Variables
and sheaved to achieve homogenize powder. Variables measured were essential amino
In this study, earthworm meal (L. rubellus) acid composition in earthworm and earthworm
was produced by using modified method from meals and also essential amino acid index in both
Edwards (1985). Earthworm sample was materials. Amino acid composition was analyzed
separated from media then washed using water by HPLC. EAAI value was calculated by Oser
and kept in the refrigerator (40C) for 12 hours. equation (1951) cit. Mitchell and Block (1946)
Formic acid (3% v/w) was added into earthworm with mathematical model:
and dried by using oven 500C for 10-12 hours.
Dried sample was grinded and sheaved become to log EAAI= 1 [log 100xA1 log 100xA2 ...log 100xAn ]
particle size + 40 mesh. Nutrient composition 10 At1 At2 Atn
proximately of earthworm meal was summarized Note:
on Table 1.
A1: first, …(1,2,3,4.......n) essential amino acid
rate from experiment sample
Analyzing Amino Acid
At1: first, …(1,2,3,4.......n) essential amino acid
Amino acid of earthworm meal was analyzed
rate from standard (whole egg)
by using High Performance Liquid
Chromatography (HPLC). Chloride acid 6N (10
RESULTS AND DISCUSSION
ml) was added into 2 gram of earthworm meal
and kept in highly temperature of disclosed
Earthworm meal L. rubellus had high protein
reaction cylinder. Sample was hydrolysed by
content and it was very important as nutrient
autoclave (1100C) for 20 hours then preserved in
source for animal. Protein content of earthworm
well-closed state at room temperature. After that,
meal was equal to fish meal (55-60% crude
sample was evaporated in 500C temperature then

254 J.Indonesian Trop.Anim.Agric. 34(4) December 2009

protein) as protein source for poultry feed. arginine, glycine, histidine, isoleucine, leucine,
Earthworm meal had also containing bioactive lysine, methionine, cystine, phenylalanine,
‘lumbricin’ in which able to inhibit pathogenic proline, threonine, tryptophan, and valine (NRC
bacteria (Cho et al., 1998). Sofyan et al. (2008) 1994). Five of them that are deemed critcial in
reported that earthworm meal with or without poultry rations are methionine, cystine, lysine,
chitosan capable to reduce E. coli colonization tryptophan, and arginine (North, 1990).
and improve protein retention in broiler chicken. Amino acid analysis with HPLC showed that
Formic acid addition (3%) into the earthworm and earthworm meal L. rubellus had
earthworm meal could increase the crude protein essential and non essential amino acid content
rate and produced low pH. The formic acid (Table 2). The highest essential amino acid of
addition into earthworm before entering the earthworm was dominated by histidine (0.63% of
grinding process was estimated could protect the dry matter basis) and earthworm meal was
earthworm meal from decomposer microbes, dominated by isoleucine (1.98% of dry matter
therefore it capable to maintain the highly protein basis). Histidine was essential for protein
content and also bioactive compound. synthesis, involved in carnitine and haemoglobin
Earthworm L. rubellus contained ‘lumbricin synthesis. It was effective in alergic disease and
I’ which had antibacterial activity, included in tension of the autonomic nervous system,
peptide group which contained 62 amino acids meanwhile isoleucine was involved in protein
(Salzet et al., 2006). Amino acid utilized as synthesis, energy production and muscle building.
proteins are primary constituents of structural and Tram et al. (2005) reported that the highest
protective tissues, including skin, feathers, bone, essential amino acid composition of P. excavatus
ligaments, as well as muscles and organs. Amino was leucine (3.47% and 0.76% of dry matter basis
acids are used as poultry feed supplement for respectively). The non essential amino acid of
optimum performance and also used in livestock earthworm and earthworm meal was dominated
health care (NOSB, 2001). As feed supplement by glutamic acid (1.52% and 3.60% of dry matter
amino acids form protein. Of the 22 amino acids basis respectively). This result also similar to
found in body proteins, the National Research previous experiments reported by Tram et al
Council lists 13 as essential in poultry diets, and (2005) that the highest non essential amino acid
these must be consumed in feed. These 13 are: composition of Perionyx excavatus was glutamic

Table 2. Amino acid composition of earthworm and earthworm meal

Earthworm meal Promin earthworm

Earthworm Earthworm meal
Amino Acid* L. rubellus L. rubellus P. excavatus 1) flour 2)
Essential
Phenyalalanine 0.45 1.03 - -
Valine 0.52 1.3 - -
Methionine 0.35 1.72 1.2 0.22
Isoleucine 0.47 1.98 2.74 0.47
Threonine 0.45 1.03 1.25 0.52
Histidine 0.63 1.91 2.87 0.26
Arginine 0.56 1.24 0.4 0.65
Lysine 0.51 1.43 0.13 0.76
Leucine 0.54 1.16 3.47 0.76
Cysteine 0.31 0.39 - -
Tyrosine 0.43 0.82 - -
Non Essential
Aspartic acid 0.98 2.38 3.18 1.16
Glutamic acid 1.52 3.6 7.13 1.42
Serine 0.54 0.7 2.45 0.5
Glycine 0.35 0.55 - -
Alanine 0.32 0.99 - -
Proline 0.54 0.71 1.81 0.53

* Dry matter basis (%); 1) Tram et al., 2005

Amino acid of earthworm meal ( L. Istiqomah et al.) 255

acid (7.13% and 1.42% of dry matter basis). EAAI value means that essential amino acid more
Glutamic acid was involved in protein synthesis balance and it could increase the biological value
and as source of energy for cells lining the of feed material, absorption and also supporting
intestine. Facilitates immune function and the poultry growth (Rama Rao et al., 1964;
improve anti-inflammatory effects, aids in Raghunath and Narasinga Rao, 1984).
preventing and healing of peptic ulcer and
ulcerative colitis.
Tabel 3. Essential Amino Acid Index (EAAI)
The high essential amino acid composition
of earthworm meal would produce great result Material EAAI Value (%)
when added in animal feedstuff ratio. By protein Earthworm L. rubellus 21.23
content rich with amino acids, the earthworm Earthworm meal L. rubellus 58.67
meal presents around 98% of absorption by Whole egg (standard) 100
animal organism due the balance between
vitamins and amino acids (Promin, 2006).
According to Table 2, the amino acid
contents of earthworm meal were varied CONCLUSION
depending on species and food source. However,
earthworms have been found to accumulate and Amino acid composition both essential and
concentrate methionine and lysine as limiting non essential of earthworm meal were higher than
amino acid found in ecosystem in proportions those of earthworm. The value of essential amino
greater than for other amino acids (Pokarzhevskii, acid index obtained from earthworm meal process
et al., 1997). Earthworm meal contains all the was higher (58.67%) than earthworm (21.23%).
essential amino acids required in feedstuff. A Powdering method of earthworm by using formic
shortage of the limiting amino acid will constrain acid had higher amino acid balance than
animal growth, reduce feed efficiency, and in earthworm.
extreme cases cause a nutritional deficiency.
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