tesis algas importante

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Review of literature

3

UTILIZATION SOME OF SEAWEEDS IN POULTRY DIETS

BY

KHALED MOHAMED EL-SYED AL-ZAABLAWYB. Sc. Agric. (Animal production), Al-Azhar University, 1999

A THESIS

Submitted in Partial Fulfillment of the

Requirements for the Degree

of

MASTER OF SCIENCE

in

AGRICULTURAL SCIENCES

(Environment and Biological Agriculture)

Department of Environment and Biological Agriculture

Faculty of Agriculture

Al-Azhar University

1426 A. H.

2005 A. D.

APPROVAL SHEET

Name: KHALED MOHAMED AL-SYED AL-ZAABLAWY

TITLE: UTILIZATION SOME OF SEAWEEDS IN POULTRY DIETS

A THESIS

Submitted in Partial Fulfillment of the Requirements

for the Degree

of

MASTER OF SCIENCE

in

AGRICULTURAL SCIENCES(Environment and Biological Agriculture)


Faculty of Agriculture, AlAzhar University

1426 A. H.

2005 A. D.

Approved by:Prof. Dr. Nohamed El-Said Farghaly..Professor of Marine Ecology, Department of Marine Science, Faculty of Science, Suez-canal University.

Prof. Dr. Abdel-hadi A. Amer..Professor of Poulltry nutriton, Department of Animal production, Faculty of Agriculture, Al-Azhar University.

Prof. Dr. Nabil N. El-Hefnawy ...Professor and Head of Environment and Biological Agriculture Department, Faculty of Agriculture, Al-Azhar University.

Prof. Dr.Khimsawy A. El-Khimsawy ...Professor of Poulltry nutriton, Department of Animal production, Faculty of Agriculture, Al-Azhar University .


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Date: 13 / 8 / 2005

TITLE: UTILIZATION SOME OF SEAWEEDS IN POULTRY NDIETS

Name:KHALED MOHAMED EL-SYED AL-ZAABLAWY

A THESIS

Submitted in Partial Fulfillment of theRequirements for the Degree

of

MASTER OF SCIENCE

in

AGRICULTURAL SCIENCES

(Environment and Biological Agriculture)


Faculty of Agriculture

Al-Azhar University

1426 A. H.2005 A. D.

Supervision Committee:-

Prof. Dr. Nabil N. El-Hefnawy ...Professor and Head of Environment and Biological Agriculture Department, Faculty of Agriculture, Al-Azhar University.

Prof. Dr.Khimsawy A. El-Khimsawy ...Professor of Poulltry nutriton, Department of Animal production, Faculty of Agriculture, Al-Azhar University .

Dr. Mohsen A. O. Elmohandes Associate Professor, Department of Environment and Biological Agriculture, Faculty of Agriculture, Al-Azhar University.

AcknowledgementI wish to express my gratitude and appreciation to Dr. Nabil N. El-Hefnawy, Professor and Head of Environment

and Biological Agriculture Department, Faculty of Agriculture, Al-Azhar University and Dr. Ayman F. Abou-Hadid,

Professor of Vegetable Crops, Department of Horticulture, Faculty of Agriculture, Ain Shams Khimsawy A. El-Khimsawy

Professor of Poultry nutrition, Department of Animal production, Faculty of Agriculture, Al-Azhar University and Dr.

Mohsen A. O. El-mohandes, associate Professor, Department of Environment and Biological Agriculture, Faculty of

Agriculture, Al-Azhar University for their supervision, suggesting the problem, valuable guidance, constructive and

fruitful help throughout writing of this dissertation.

My deep thanks are also due to and Dr. Mahmoud El-Said Farghaly, Professor of Marine Ecology, Department ofMarine Science, Faculty of Science, Suez-canal University.

Many thanks are also due to all staff members of the Department of Environment and Biological Agriculture,Faculty of Agriculture, Al-Azhar University and all staff members of the Department of Animal production, Faculty of

Agriculture, Al-Azhar University.

Special thanks are due to my family who encouraged and pushed me forward to achieve the present work.

Finally my deep thanks are due to all people who taught and directed me throughout my life.


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Table of Contents

page1- Introduction. 1

2- Review of Literature... 3

2.1. Algae and Seaweeds 3

2.2.1. The major components of Seaweeds (ash,

protein, fiber, fat, and carbohydrates). 162.2.2. Amino acid and fatty acid composition of

seaweeds. . 22

2.2.3. Mineral and heavy metals contamination in theseaweeds. .. 27

2.2.4. Vitamins in seaweeds. 31

2.3. Effect of seasons on seaweeds. 33

2.3.1. The seasonal effects on yield and gel propertiesof seaweeds. 33

2.3.2. Seasonal effects on chemical composition of theseaweeds. 36

2.4. Cell wall degradation by chemical and enzymatic

treatment for improvement of protein extractionfrom seaweeds. 41

2.5. Seaweeds in poultry feeding. . 46

3- Materials and Methods...... 54

4- Results and Discussion..... 61

4.1. Effect of area and season on chemical composationof algae. .. 61

4.2. Degradation of algal cell walls to improving proteinaccessibility. . 86

4.3. Effect of chemical and enzymatic treatments ondigestibility of nutrients on Japanese Quail.. 105

5- Summary and conclusion.. 110

6-References.... 116

7- Arabic Summary...

LIST OF TABLES

Table page1 Seaweed world production (Tons) 132 Production and value of international seaweed gums market,

1995. . 153 The nutritional composition of Green Seaweeds(Chlorophyta) 204 The nutritional composition of Red Seaweeds (Rhodophyta)... 215 Free Amino Acid Composition (mg/gm protein) of several

species of seaweeds. Ito & Hori (1989) 226 Amino acid composition (mean % SE) of the algae

Chlorophyta and Rhodophyta . Wahbeh (1997). .. 237 Fatty acid composition (mean % SE) of the algae,U.

lactucaand P. pavonicafrom Aqaba.Wahbeh (1997) 248 Free Amino Acid Compsition (mg/gm protein) of several

species of seaweeds. . 249 Examples of ash and mineral contents in some Seaweeds. .. 28

10 Mineral composition of Seaweeds. (mg/g dry matter).. 3011 seasonal changes the constituents ofUlva lactuca. .. 3712 Amino acid composition ofUlva amoricanasamples

collected in October 1997, December 1997 and February1998 (g 100 g

-1protein). ... 39

13 Composition of basel diet. .... 5714 Chemical analysis of group diets were studied.. 5815 Content ofUlvafrom moisture. ... 6216 Content ofUlvafrom cured protein. 6417 Content ofUlvafrom ether extract. .. 67


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18 Content ofUlvafrom crude fiber. 6919 Content ofUlvafrom ash %. 7220 Content ofUlva from N. F. E. % . 7421 Content ofGelidium from moisture. 7622 Content ofGelidium from cured protein. 7823 Content ofGelidium from ether extract. ... 7924 Content ofGelidium from crude fiber. . 8125 Content ofGelidium from ash %. . 82

26 Content ofGelidium from N. F. E. % . . 8427 Effect of acetic acid treatment on seaweeds 8728 Effect of sodium hydroxide treatment on seaweeds. 9129 Effect of calcium hydroxide treatment on seaweed. . 9330 Effect of acetic acid and sodium hydroxide treatment on

seaweeds. .. 9731 Effect of enzymes on seaweeds. 9932 Effect of all previous treatments on seaweeds.. 10333 Effect of treatments on digestibility of crude protein (CP),

ether extract (EE) and N-free extract (NFE) and (DE)digestible energy of seaweeds. 105

LIST OF FIGURESFigure page

1 some genus of Clorophyta ( green algae) 7

2 some species of Ulva sp. 83 some genus of Rhodophyta (red algae). . 94 some species of Gelidium.

105 Content ofUlvafrom moisture. ... 636 Content ofUlvafrom cured protein. 657 Content ofUlvafrom ether extract. .. 688 Content ofUlvafrom crude fiber. 709 Content ofUlvafrom ash %. 73

10 Content ofUlva from N. F. E. % . 7511 Content ofGelidium from moisture. 7712 Content ofGelidium from cured protein. 7913 Content ofGelidium from ether extract. ... 8014 Content ofGelidium from crude fiber. . 8115 Content ofGelidium from ash %. . 8316 Content ofGelidium from N. F. E. % . . 8417 Effect of acetic acid treatment on seaweeds 8818 Effect of sodium hydroxide treatment on seaweeds. 9219 Effect of calcium hydroxide treatment on seaweed. . 9420 Effect of acetic acid and sodium hydroxide treatment on

seaweeds. .. 9821 Effect of enzymes on seaweeds. 10022 Effect of all previous treatments on seaweeds.. 10423 Digestion trails. 10624 Digestible energy. 108

1. INTRODUCTIONSeaweeds can be classified into three broad groups based on pigmentation: brown, red and green. Botanists refer to these broad groups as Phaeophyceae,

Rhodophyceae and Chlorophyceae, respectively. Brown seaweeds are usually large, and range from the giant kelp that is often 20 m long, to thick, leather-like seaweedsfrom 2-4 m long, to smaller species 30-60 cm long. Red seaweeds are usually smaller, generally ranging from a few centimetres to about a metre in length. Greenseaweeds are also small, with a similar size range to the red seaweeds.

Seaweed is a very versatile product widely used for industryal. Red and brown seaweeds are also used toproduce hydrocolloids alginate, agar and carrageenan, which are used as thickening and gelling agents. Today,approximately 1 million tonnes of wet seaweed are harvested and extracted to produce about 55 000 tonnes ofhydrocolloids, valued at almost US$ 600 million.(Mc.Hugh, 2003).

Seaweed used for food in direct human consumption and animal. It is also an ingredient for the global food and cosmetics industries and is used as fertilizer and as ananimal feed additive. Total annual value of production is estimated at almost US$ 6 billion of which food products for human consumption represent US$ 5 billion. Totalannual use by the global seaweed industry is about 8 million tonnes of wet seaweed.

Use of seaweed as food has strong roots in Asian countries such as China, Japan and the Republic of Korea, but demand for seaweed as food has now also spread toNorth America, South America and Europe.

However, only afew studies have been undertaken on the quality of seaweed proteins because the extraction of protein from seaweed is difficult because of theoccurrence of phenolic compounds and large amounts polyanionic cell wall mucilages (Fleurenceet al.1995).


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The first part of present study was observe undertaken to ascertain whether seasonal variation occurs in the chemical composition of seaweeds (Gelidiumsp. ,Ulvasp.)on north coastal in Egypt.

The second part was investigated the degradation of cell wall polysaccharides of saewweds by several chemical and enzymatic methods to improving the extraction ofproteins.

In the last determinated digestibility of the protein extraction by chicken.

2. REVIEW OF LITERATURE

2.1. Algae and Seaweeds

Algae are very simple chlorophyll-containing organisms: some say that they are plants other says that are not,calling them Protists. We use the term "algae" very loosely because defining them is very difficult. In general,we can say that they are simple organisms composed of one cell, or grouped together in colonies, or asorganisms with many cells, sometimes collaborating together as simple tissues.2.1.1. Classification of algae

According toLee (1999)classification of algae. Groups are based on the number of chloroplast membranes.Group 1. Prokaryotic Algae, Ph. Cyanobacteria no chloroplasts (1 membrane). Group 2. Eukaryotic Algae,chloroplasts with 2 membranes Ph. Glaucophyta, Ph. Rhodophyta (red algae) and Ph. Chlorophyta (greenalgae). Group 3. Eukaryotic algae chloroplasts with 3 membranes, Ph. Euglenophyta (euglenoids) and Ph.Dinophyta (dinoflagellates). Group 4. Eukaryotic algae chloroplasts with 4 membranes, Ph. Cryptophyta(cryptophytes), Ph. Heterokontophyta (heterokonts), (Chrysophyceae, Synurophyceae, Dictyochophyceae,Pelagophyceae, Bacillariophyceae (diatoms), Raphidophyceae, Xanthophyceae, Eustigmatophyceae,Phaeophyceae), and Ph. Pymnesiophyta.

2.1.1.1. Chlorophyta (green algae)The green algae (chlorophyta) are probably the most strucurally diverse grup of algae with many types ofunicells, colonies, filaments, siphons, and thalloid forms. Recent ultrastructural studies have established fourlines of evolution from the primitive unicellular condition(Mattox and Stewart 1984). The chlorophyceae are

primarily freshwater forms this class contains most of the species of green algae includingChlamydomonasand the volvocine line,Chlorellaand most other chlorococcalean forms, a number of branched and unbranchedfilaments, and the familiar Oedogoniales. Most of the marine green algae belong to the Ulvophyceae, agenerallytropical and subtropical group containing a number of relatively large forms (e.g.,Ulva, Codium, Valonia,

Halimeda), even though Plasmodesmata (a prerequisite for tissue diferentiation and specialization) neverevolved in this class(Graham 1984).Figure (1) show some of green algae.*Sea Lettuce (Ulva): it known by the common name sea lettuce,Ulvacan be eaten in salads or used in soups.Ulva is a particularly popular food in Scotland. Nutritionally, it is very healthy. U. lactucais made of 15%

protein, 50% sugar and starch, less than 1% fat, and 11% water when dried. It is useful as roughage in thehuman digestive system. Ulvaare very high in iron, as well as high in protein, iodine, aluminum, manganeseand nickel. They also contain vitamin A, vitamin B1, vitamin C, sodium, potassium, magnesium, calcium,soluble nitrogen, phosphorous, chloride, silicon, rubidium, strontium, barium, radium, cobalt, boron and traceelements.Ulvaspecies have thalli with expanded blades two cells thick (distromatic). Ulvaare

parenchymatous: cell division may occur anywhere on the thallus but always in a plane perpendicular to thethallus surface. Compared to more advanced algae and vascular plants, their construction is relatively simple.They do not differentiate into tissue layers or show much specialization among cells. The cells themselves areirregularly arranged and are quadrate to slightly elongate anticlinally (perpendicular to the surface), dependingon the species. The cell walls are fibrillar and made up of cellulose. They store energy as starch. Arranged insheets only two cells thick,Ulvas large surface to volume ratio allows it to have a high nutrient uptake. Ulva

generally lives in the middle to low intertidal zone (or eulittoral to high sublittoral zone). The fronds are notsituated at the same level throughout the year, however. In the colder months, the algae grow mainly in wide

bands in the intertidal. In the warmer months, they grow in a narrower band, lower in the intertidal.Minimizing the amount of time they spend out of the water, under the hot summer sun, protects them fromdesiccation. Ulvaare greatly impaired by extreme desiccation (defined as loss of more than 25% original watercontent). Figure (2) show some of Ulva sp.

2.1.1.2. Phylum Rhodophyta (red algae):The red algae (Rhodophyta) are the third group (along with the brown and green algae) that contributes to

the seaweed flora. Some red algae are microscopic and even unicellular. A few grow in swift currents infreshwater streams(Sheat and Hymes 1980)but are often overlooked due to their small size. Most, however,are conspicuous, multicellular seaweeds. Carrageenan and agar are commercially valuable polysaccharidesobtained from certain red algae. The earlier theory that the pigmentation in red algae was a special adaptation tothe low irradiances in deep water has now been disputed (Ramus 1983).Figure (3) show some of red algae,figure (4) show some species ofGelidium.


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Figure (1) .some genus of Clorophyta ( green algae)

Figure (2). some species of Ulva sp.

Ulva

Codium Caulerpa

LactucaUlva PertusaUlva

RigidaUlva

LobataUlva

Valonia


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Figure (3).some genus of Rhodophyta (red algae).

Figure (4) . some species of Gelidium.

*Gelidium sp.The genus Gelidiumbelongs to the family Gelidiaceae, which also includes eight other genera. Thefamily Gelidiaceae has been considered a member both of the order Gelidiales and of the Nemalionales(=Nemaliales). The classification of the family depends upon acceptance of the Gelidiales as a distinct order, ataxonomic status which has been under dispute over the last twenty years(Dixon, 1961 Pueschel and Cole,1982).

Species ofGelidiumare among the most important agarophytes in the world(Santelices, 1974 Santelicesand Stewart, 1985). About 35 species are harvested in various areas contributing to 40-50% of the world'sannual exploitation of agarophytes, estimated at 39,000 tons of dry matter(Whyte and Englar, 1981).2.1.2. World production and uses of seaweed.

2.1.2.1. World production of seaweed:Between 1981 and 1994 world production of seaweed increased from 3.2 million tonnes (fresh weight) to

nearly 7 million tones. The seaweeds that are most exploited are the brown algae with about 5.2 million tones

(75%) followed by the red algae (1.73 million t 25%) and a small amount of green algae (about 0.5%).World production of seaweed grew by 99.37% during the period 1993-1997. As reported by Food andAgriculture Organization (FAO) during the same period, the Philippines ranked 5th among the major producingcountries of seaweed. The Philippine contributed 6.56% to the total world production of 559,888,073 MT(Table 1). China, on the other hand is the major producing of seaweed with 292,441,630 MT which contributed

Gracilaria Porphyra

Gelidium

CoulteriGelidium PusillumGelidium

PacificumGelidium


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52.23% or more than half of the world production. Second producing country is Korea D P Rp with 57,221,136MT followed by Korea Rep with 50,027,596 MT. Their total contribution was 10.22% and 8.94% respectively.

2.1.2.2. Uses of Seaweeds:

2.1.2.2.1. Human food, animal fodder and manure:Seaweed as a staple item of diet has been used in Japan and China since prehistoric times. In 600 BC, Sze Teu wrote in China, "Some algae are a delicacy fit for the most

honored guests, even for the King himself." Some 21 species are used in everyday cookery in Japan, six of them since the 8th century. Seaweed accounts for some 10% ofthe Japanese diet and seaweed consumption reached an average of 3.5 kg per household in 1973, a 20% increase in 10 years (Indergaard and Minsaas 1991).


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Table (1): Seaweed world production (Tons)All Fishing

Areas1993 1994 1995 1996 1997 % Share

Growth Rate(1993-1997)

Total WorldProduction

36,566,591 71,624,400 141,389,256 280,902,907 559,888,073 100.00 -

China 18,511,176 36,821,085 73,346,026 146,377,635 292,441,630 52.23 99.37

Korea D P Rp 3,594,446 7,166,892 14,316,784 28,618,568 57,221,136 10.22 99.75

Korea Rep 3,352,172 6,464,342 12,650,014 25,098,883 50,027,596 8.94 96.57

Japan 3,411,847 6,453,944 12,418,978 24,426,747 48,476,658 8.66 94.18

Philippines 2,704,308 5,031,093 9,606,416 18,666,506 36,720,567 6.56 92.00Chile 1,241,142 2,403,421 4,702,937 9,243,340 18,325,732 3.27 96.03

Norway 904,545 1,809,090 3,618,180 7,236,360 14,472,720 2.58 100.00

Indonesia 673,951 1,229,507 2,348,576 4,585,577 9,009,611 1.61 91.29

USA 405,972 811,944 1,623,888 3,247,572 6,494,882 1.16 99.99

India 437,800 831,500 1,617,900 3,190,700 6,334,700 1.13 95.06

Others 1,329,232 2,601,582 5,139,557 10,211,019 20,362,841 3.64 97.84

Source: FAO Statistics.


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The most important food species in Japan are Nori (Porphyra species), Kombu (Laminaria species), and Wakame

(Undaria pinnatifida). In the west, seaweed is largely regarded as a health food and, although there has been an upsurge

of interest in seaweed as food in the last 20 years, it is unlikely that seaweed consumption there will ever be more than a

fraction of the Japanese.

Human consumption of Gelidium is restricted mainly to G. divaricatum in China and to G. amansii in Japan,

Indonesia, China, Borneo and The Celebes Zaneveld, 1955, 1959 Johnston, 1966 Levring et al. 1969). Nowhere are

the species used as animal fodder or manure. In Chile, all the harvested crops are exported as raw materials for agar

production.

2.1.2.2.2. Industrial products and processes

Industrial gums extracted from seaweeds fall into three categories: alginates (derivatives of alginic acid), agars and

carrageenans. The first is extracted solely from brown seaweeds whilst the last two are extracted only from red seaweeds.

There are a number of artificial products reputed to be suitable replacements for seaweed gums but none have the exact

gelling and viscosity properties of seaweed gums and it is very unlikely that seaweeds will be replaced as the source of

these polysaccharides in the near future.

Production and value of international seaweed gums market, 1995 showed in Table (2).

Table (2):production and value of international seaweed gums market, 1995.Seaweed gum Total (t) Price ($ per kg) Total value ($ million)

Agar 10,161 20 203

Carrageenans 25,403 8 203

Alginates >25,000 6 150

Total >61,000 - 560

Sources: Quest International, Cork and IMR International, San Diego).

Species of Gelidium are among the most important agarophytes in the world (Santelices, 1974 Santelices and

Stewart, 1985). About 35 species are harvested in various areas contributing to 40-50 % of the world's annual

exploitation of agarophytes, estimated at 39,000 tons of dry matter (Whyte and Englar, 1981). However, agar

concentrations found in several Gelidium species Gelidium chilense produces the highest agar yield while G. lingulatum

yields the least gel (20%). Gelidium rexyields an intermediate amount of gel but it produces the strongest gel(Santelices,

Oliger & Montalva 1981).

Alginates are cell-wall constituents of brown algae (Phaeophycota). They are chain-forming heteropolysaccharides

made up of blocks of mannuronic acid and guluronic acid. Composition of the blocks depends on the species being used

for extraction and the part of the thallus from which extraction is made.

Carrageenan is a general name for polysaccharides extracted from certain kinds of algae which are built up, in

contrast to agar, from D-galactopyranose units only. The use of this seaweed to extract a gel is known in Ireland since

1810. Chondrus crispus used to be the sole source of carrageenan, but species of Gymnogongrus, Eucheuma, Ahnfeltia

andGigartinaare now commonly used.

2.2.1. The major components of Seaweeds (ash, protein, fiber, fat, and carbohydrates).Castroet al. (1991)found that the nutritional composition of washed and unwashed sun-dried seaweed (Macrocystis pyrifera) meal was evaluated by chemical analysis

and in vitro and in situ digestibility. In the unwashed and washed meal, nitrogen free extract comprised 46.27 and 46.67%, respectively and ash 36.67 and 34.22%.Washing significantly increased the content of some minerals in the seaweed. Although protein percentage was low (8.8%), it had a good amino acid balance. Tannin was

only detected at a low level (34.20 mg/g). In vitro and in situ DM digestibility were high (90.34 and 83.24%, respectively). It is concluded that Macrocystis pyrifera can be

included in animal feeds, and that prior washing is not necessary.

Lahaye (1991)studied the soluble and insoluble dietary fiber contents of marine algae ('sea vegetables'), wakame (Undaria pinnatifida), hijiki (Hijiki fusiformis), seaspaghetti (Himanthalia elongata), arame (Eisenia bicyclis), sea lettuce (Ulva lactuca), A O nori (Enteromorpha spp.), and nori (Porphyra tenera) were estimated by the

gravimetric method ofProskyet alwith adaptations. These seaweeds had total dietary fiber contents between 32.7 and 74.6% (on a dry weight basis) of which 51.6 to85.0% were water soluble.


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Lahaye and Jegou(1993)studiedUlva lactucacontained 15.8 and 8.0% soluble fibres according to the standard and physiological methods, respectively, and24.2 and 32.6% insoluble fibres according to the 2 methods, respectively. ForEnteromorpha compressathese values were 14.9 and 15.9%, and 21.6 and 28.7%,

respectively. For both algae, soluble fibres appeared to be xylorhamnoglycuronan sulphates and insoluble fibres were essentially composed of glucans. Fibres in both algaewere hydrophilic but the water-holding capacities were higher after extraction of soluble fibres. Water-soluble fibres, particularly those from E. compressa, demonstrated

low intrinsic viscosities at 37C in buffers and were affected by pH.

Ventura et al. (1994)studied nutritive value of seaweed (Ulva rigida) in poultry diets and found that proximate composition of seaweed was nitrogen 33, crude fibre 17,neutral detergent fibre 312, acid detergent fibre 153, pentosans 13 and ash 228 g/kg DM.

Robledo and Freile (1997)determinedash, protein, fibre, fat, carbohydrates in samples ofGracilaria cornea,Eucheuma isiforme,Caulerpa racemosa,Codiumisthmocladum,Padina gymnosporaandSargassum filipendulaand they found the ash contents ranged from 29.06 to 55.93%.E. isiformehad the highest protein content

(12.10%), while lowest value was inC. isthmocladum(3.50%). Fat content was highest inCodium isthmocladumandGracilaria cornea(0.48 and 0.26%, respectively). ).Crude fibre varied from 1.01 to 9.07%.E. isiformeandG. corneahad the highest carbohydrate contents (25.89 and 36.29%, respectively).

Ventura and Cast-Anon (1998)studiedthe nutritive value of seaweed in adult male Canarian goats. The nutrient content ofU. lactuca, per kg DM, was 825 g organicmatter (OM), 211 g CP, 17 g ether extract, 189 g structural carbohydrates, 27 g lignin and 381 non-structural carbohydrates. Effective rumen degradation was OM 335 andCP 96 g/kg DM, and in vitro digestion was OM 512 and CP 147 g/kg DM. The energy content of seaweed was estimated to be 10.2 MJ DE/kg DM.

Wong and Cheung (2000)investigated the proximate composition, amino acid profile and some physico-chemical properties of two subtropical red seaweeds (HypneacharoidesandHypnea japonica) and one green seaweed (Ulva lactuca) were investigated. The total dietary fibre [ranged from 50.3 to 55.4% dry weight (DW)] and ash

(ranged from 21.3 to 22.8% DW) were the two most abundant components in these seaweeds but their crude lipid contents were very low (ranged from 1.42 to 1.64%DW).

Tables (3, 4) show the different chemical composition of some seaweed (green & red seaweeds).


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Table (3). The nutritional composition of Green Seaweeds. (Chlorophyta).

Authors SeaweedsAsh

% dryProtein

% dry

Crbohaydrate

% dryFiber% dry

Faty acid

% dry

Abdel-fattah & Edrees

(1972) Ulva.lactuca

24.03

35.89

8.70

33.75 _ _

2.91

3.94Ito & Hori (1989) Ulva.lactuca 18.7 15.2 39.1 4.3 0.6

Mohammed (1997) Ulva.lactuca33.3 +

6.417.63.1

_ _ 5.2 1.2

Greg &Alan (1998) Ulva.rigida4752

6.45.9

18.117.3

_ 0.30.6

Wong & Peter (2000) Ulva.spp. 21.3 7.06 14.60 55.4 1.64

David (2001) Ulva.spp. 14 24 47 1.00 _


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Table (4). Chemical composition of Red Seaweeds (Rhodophyta).

Authors SeaweedsAsh

%dryProtein% dry

Crbohaydrate%dry

Fiber%dry

Faty acid% dry

Ito & Hori (1989)Gracilaria.spp.

10.3 9 55.8 8.3 0.1

Ito & Hori (1989) Porphyra.spp. 6.9 38.8 39.5 1.8 1.9

Ito & Hori (1989)Gelidium

amansii

_ 12.8 _ _ _

Greg &Alan

(1998)

Gracilaria

plstoldes1416

10.811.8

41.443.1

_0.91.2

Greg &Alan(1998)

Corallina.spp7780

6.46.1

4.24.7

_0.71.1

David (2001)Gracilaria.spp

.17 11 54 _ 03


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2. 2.2. Amino acid and fatty acid composition of seaweeds.Ito & Hori (1989)studied content of several species of seaweeds (U. lactuca, E. compressa, P. pavonic and L. obtuse) from free amino acids and the results are shown in

Table (5).

Table (5): Free Amino Acid Composition (mg/ gm protein) of several species of seaweeds.Ito & Hori (1989)

Amino Acid U. lactuca E. compressa P. pavonica L. obtuse

Glycine

AlanineValineLeucine

IsoleucineMethionine

PhenylalanineProlineSerine

ThreonineCystine

TyrosineAspartic acid

Glutamic acidLysine

ArginineHistidine

2.2 0.4

1.80.35.60.85.01.12.80.2

19.02.412.90.3

-5.4 1.25.6 0.8

-2.6 0.29.20.63.2 0.416.70.83.0+0.34.60.7

3.3 0.5

3.0 0.65.3 1.24.6 0.73.3 0.813.80.41 l.02.72.00.22.3 0.42.7 0.65.3 1.15.70.85.4 1.26.2 1.34.4 1.13.00.38.5 1.2

1.10.2

5.85.94.87.87.69.0_

8.40.89.6

4.1 0.4.7

5.2 0.33.7 0.6

7.04.810.4

1.1 0.1

1.1 0.23.7 1.96.8 1.44.9 1.2

-5.3 0.59.1 1.59.4 1.59.7 2.5

-4.90.64.1 2.812.43.5

13.3 2.75.8 1.18.4 1.7

Wahbeh (1997)studied content of seaweeds chlorophyta (Ulva pertusandEnteromorpha liza) and Rhodophyta (Propheyra spp. andGracilaria compressa) from amino acidsand the results are shown in Table (6).

Table (6).Amino acid composition (mean % SE) of the algae, chlorophyta and Rhodophyta, Wahbeh (1997).

Wahbeh (1997) studied content of seaweeds (U. lactucaandP. pavonic)from fatty acids and the results are shown in Table (7).

Table (7).Fatty acid composition (mean % SE) of the algae,U. lactucaandP. pavonicafrom Aqaba.Wahbeh (1997).

Chlorophyta RhodophytaAmino acid Ulva

pertusa

Enteromorpha

linza

Porphyra

spp.

Gracilaria

compressaAlanine 17.7 23.8 1750 4.9

Arginine 2.5 1.9 11 26.3Aspartic acid 4.4 13.7 310 6.1

Chondrine - - - -Citrulline 28.7 43.0 - 66.2

Glutamic aci 31.8 55.0 1378 13.3

Glycine 9.1 5.2 20 TraceHistidine _ - 16 -

Isoleucine 3.7 5.8 16 TraceLeucine 6.6 6.8 35 -

Lysine 0.9 0.8 12 -Methionine - - 5 -

Phenylalanine 4.1 3.8 - -Proline 40.0 51.1 17 -Serine 11.9 33.7 41 Trace

Taurine 154.0 75.4 1569 58.7Threonine 5.7 3.8 38 Trace

Tyrosine 2.1 2.1 4 -Valine 3.5 3.9 37 -


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Percentage of total fatty acidFatty acid

U. lactuca P. pavonica

Saturated

12:014:015:016:0

18:02o:0

Monounsaturated14:1(-9)16:1(-9)16:1(-7)

18:1 (-9)18:1(-5)20:1(-9)

Polynsaturated16:2( - 6)

16:2(-9)16:3(-6)16:3(-3)16:4(-3)18:2(-9)18:2(-6)18:3(-6)

I8:3( - 3)18:4( - 3)20:2( - 6)

1.30.21.50.2

1.1 0.33.20.3

1.40.16.5 0.4

4.8 0.4-

3.6O.26.9 0.6

1.30.32.l0.4

20.31.6-

0.50.1-

6.10.49.4 0.89.7 0.6

5.60.57.1 0.74.6 0.33.00.2

-2.9 0.21.40.2

4.1 0.36.3 0.4

8.70.6

-1.50.3

-18.5 0.8

1.80.44.00.5

15.71.13.00.2

-3.10.3

-6.80.5

6.7 0.6-

5.8 0.43.8 0.45.3 0.6

Fleurence (1999)studied content of two species of seaweeds (Ulva amoricanaandpropheyra tenra) from amino acids, alsoWong and Peter (2000)studied content ofanther two species of seaweeds (U. lactucaandHypenea charoides) from the same amino acids and the results are shown in Table (8).

Table (8).Free Amino Acid Composition (mg/ gm protein) of several species of seaweeds.

Amino acid Ulva lactuca**Ulva

amoricana*Hypnea

charoides**Porphyra

tenra*

Alanine 96.7 5.5-0.7 60.6 7.4Arginine 48.6 4.3-8.7 98.1 16.4

Aspartic acid 139 6.0-11.8 163 7.0Cystin - - - -

Glutamic acid 110 11.7-23.4 125 7.2Glycine 65.3 6.3-7.5 55.2 7.2

Histidine 13.1 1.2-2.1 7.67 1.4Isoleucine 40.0 2.3-3.6 39.2 4.0

Leucine 72.6 4.6-6.7 69.8 8.7Lysine 46.4 3.5-4.4 39.2 4.5

Methionine 6.12 1.4-2.6 16.2 1.1Phenylalanine 57.1 5.0-7.1 42.2 3.9

Proline 45.7 5.0-10.5 35.3 6.4Serine 62.8 5.6-6.1 46.8 2.9

Threonine 62.0 4.5-6.8 48.3 4.0Tyrosine 36.3 4.4-4.7 29.1 2.4

Valine 52.6 4.0-5.2 52.1 6.4* Fleurence.J. (1999)

** Wong & Peter (2000)

Wong and Cheung (2000)investigated the proximate composition, amino acid profile and some physico-chemical properties of two subtropical red seaweeds (HypneacharoidesandHypnea japonica) and one green seaweed (Ulva lactuca). Although the crude protein content of the red seaweeds was significantly (p < 0.05, ANOVA, Tukey-

HSD) higher than that of the green, the three seaweeds' proteins contained all essential amino acids, the levels of which were comparable to those of the FAO/WHOrequirement. Moreover, the swelling capacity (SWC), water-holding capacity (WHC) and oil-holding capacity (OHC) of the seaweeds had a high positive correlation (r=0.99-

1.00) with their total amount of fibre and protein. Mohdet al. (2000)determined Composition ofG. changgi[C. changii].It contained a higher composition of unsaturated fatty acids (74%), mainly omega-fatty acids and

26% of saturated fatty acids (mainly palmitic acid) and also relatively high levels of calcium and iron. Major amino acid components were glycine, arginine, alanine andglutamic acid. Among the essential amino acids, lysine with a chemical score of 53% was the most limiting when compared with the essential amino acid pattern of egg

protein. This study was conducted to create nutritional data forG. changgiin order to popularize its consumption and utilization in Malaysia. Comparisons with nutritive

value of several commonly consumed local vegetables are made.

Wong and Cheung (2001)evaluated the nutritional values of seaweed protein concentrates (PCs) isolated from two red seaweeds (Hypnea charoidesandHypneajaponica) and one green seaweed (Ulva lactuca) by determining their in vitro protein digestibility and amino acid profiles. Both protein extractability and in vitro protein

digestibility of the red seaweed PCs (88.7-88.9%) were significantly (P < 0.05, ANOVA, Tukey-HSD) higher than those of green seaweed PCs (85.7%). The total amount ofessential amino acids (EAAs) in the three seaweed PCs was high (36.2-40.2% of total amino acid content). All three seaweed PCs were rich in leucine, valine and threonine

but lacked cystine. However, except for sulfur-containing amino acids and lysine, the levels of all EAAs were higher than those of th e FAO WHO requirement pattern.


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24

2.2.3. Mineral and heavy metals contamination in the seaweeds.Lacardeet al.(1985)studied the ash contents of seaweeds. They found the ash was vary widely from 8% to 40%, dry weight. All minerals required by humans: calcium,sodium, magnesium, potassium, phosphorus, iodine, iron and zinc are present in sufficient quantities. Some examples of ash and mineral contents are shown in Table (9).


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28

Table (9). Examples of ash and mineral contents in some Seaweeds.

Seaweed Total ash(% DM)

Mineral (mg/g dry matter)

K Ca Mg Si P Sr Fe Al Zn B

Clorophyta

Monostroma nitidum 22.5 .9 7.6 13.6 13.6 - 0.90 0.18 0.90 1.15 0.21 0.050

Ulva pertusa 18.8 6 5.1 8.0 25.8 5.2 1.45 0.22 0.76 0.56 0.14 0.065U. conglobata 15.9 0 1.7 8.3 36.5 - 0.80 - 0.61 1.01 0.06 -

Enteromorpha compressa 22.6 - 11.9 19.8 21.8 - 0.33 1.13 0.69 0.25 0.116

Chaetomorpha crassa 11.3 5 13.2 10.3 11.3 - 0.80 0.23 0.36 0.56 0.15 0.140

Phaeophyta

Padina arborescens 14.2 .1 35.7 19.2 8.0 - 0.98 1.31 0.84 0.77 0.14 0.130

Ishige foliacea 14.7 - 12.4 9.1 13.9 - 1.17 0.41 0.14 0.17 0.073

Scytosiphon lomentaria 23.5 9 8.7 31.9 10.7 2.5 1.80 - 1.49 1.40 0.19 0.053

Eisenia bicyclis 14.9 .8 22.1 15.0 9.0 - 0.90 1.10 0.10 0.10 0.10 0.097

Hizikia fusiforme 20.6 .9 34.3 17.2 9.9 - 0.90 1.14 0.16 0.20 0.08 0.109

Sargassum ringgoldianum 14.4 2 17.4 20.8 10.7 - 0.75 1.48 0.11 0.10 0.06 0.097

S. tortile 13.5 7 5.4 28.8 8.7 - 0.70 1.98 0.11 0.08 0.17 0.066

S. thunbergii 20.7 8 20.9 26.2 9.7 6.8 1.10 1.65 0.71 1.03 0.32 0.122

Rhodophyta

Gelidium amansii 7.7 2 0.3 7.0 5.3 3.3 0.90 0.09 0.36 0.14 0.16 0.177Acanthopeltis japonicus 12.1 8 1.8 1.1 4.8 - 1.05 0.02 0.23 0.32 0.17 -

Carpopeltis labellate 13.8 10.7 6.5 3.8 7.8 - 1.20 - 0.30 0.24 0.17 0.055

Gloiopeltis tenax 11.5 - - 5.1 3.3 - - 0.03 0.16 - 0.08 0.034

Gymnogongrusflabelliformis

12.5 - - 2.8 4.2 - - 0.09 0.28 - 0.12 0.217


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RESULTS ND DISCUSSION

61

Ito and Hori (1989).Studied minerals content of seaweeds ( ulva pertusa). Table (10) indicate the results he obtained.Seferlis and Haritonidis (1995)studied three salinity values (25%, 30% and 36%) correspond to three sites in the Thermaikos Gulf, Thessaloniki, each under the influence

of different pollution loadings (industrial, domestic, agricultural run-off). During 1992 the content of Pb, Cu and Cd in the thalli of the green alga were monitored at fourteenstations along the coastline of the gulf. Pb ranged from 29ppb to 119ppb, Cd from 129ppb to 448ppb and Cu from 3ppb to 180ppb. The experiment was designed so that

specimens could be classified by two characteristics (salinity - time and metal enrichment). Special attention was given to which combination of salinity and additional metal

loading, produces a very small or a very large accumulation. The effect of each variable was tested separately.

Robledo and Freile(1997)determined Na, K, Mg, Ca, Fe, Cu, Cr, Zn and Pb in samples ofGracilaria cornea,Eucheuma isiforme,Caulerpa racemosa,Codiumisthmocladum, Padina gymnosporaandSargassum filipendula. S. filipendulaandP. gymnosporawere characterized by a high mineral content Ca and Mg were particularly

high. Trace metals (Fe, Pb, Cu, Cr and Zn) occurred at high levels in all species. David (2001).Studied minerals content of Rhodophyta ( Gracilaria). Table (10) indicate the results he obtained.

Table (10).Mineral composition of Seaweeds. (mg/g dry matter).

Clorophyta Rhodophyta

Minral Ulva pertusa* Gelidium amansii* Gracilaria**

Na 3.6 7.7 53.4

K 5.1 0.2 -

Ca 8.0 7.0 41.00

Mg 25.8 5.3 -

Si 5.2 3.3 -

P 1.45 0.9 60.00

Fe 0.76 0.36 0.21

Al 0.56 0.14 -

Zn 0.14 0.16 0.041

B 0.065 0.177 -

I 0.31

Cu 0.008

Cr 0.006

*Ito & Hori (1989)** David (2001)

Calicetiet al.(2002)evaluated the concentrations of heavy metals (Fe, Zn, Cu, Cd, Ni, Pb, Cr, As) in seven seaweeds of environmental and commercial relevance (UlvarigidaC. Ag.,Gracilaria gracilis(Stackhouse)Steentoft,L. IrvineandFarnham,Porphyra leucosticta Thuret, Grateloupia doryphora(Montagne) Howe.,Undaria

pinnatifida (Harv.) Suringar, Fucus v irsoides J. Agardh, Cystoseira barbata (Good. et Wood. Ag) collected in four sampling sites in the lagoon of Venice, in spring andautumn 1999. Metals were extracted using hot concentrated acids in a Microwave Digestion Rotor and analysed by absorption spectrophotometry using a flame mode for Feand Zn and a graphite furnace for Pb, Cr, Cd, Cu, Ni and As. High contamination levels, especially for Pb, were detected in Ulvaand to a lesser extent inGracilaria. Brown

seaweeds,especially Cystoseirawas highly contaminated by As. The least contaminated genera with all metals except As werePorphyraandUndaria. A concentrationdecrease for Zn and Cd was observed from the inner parts of the central lagoon, close to the industrial district, towards the lagoon openings to the sea.

2.2. 4. Vitamins in seaweeds.Jensen (1963)surveyed 25 different species of Norwegian seaweeds for tocopherols and reported that tocopherol is persent (7-220ppm) in all classes of seaweeds

examined. In genral, green, red, and sublittoral brown seaweeds seem to be rather poor in vitamin E. Kanazawa (1963)reviewed distribution of vitamins in seaweeds. Seaweeds, as well as green vegetables, contain all kinds of vitamins and are a natural source of

vitamins for humans.Enteromorphassp. andPorphyrassp. Contain larger amounts of B-carotene (pro-vitamin A) than yello-green vegetables. Amounts rang from 22 to 25mg per 100 g of air dried materils.

Hedetermined the composition of vitamin B in 35 different specis of Japanese seaweeeds. The results showedthe Porphyra spp.flavein adenine dinucleotide occupies over 90% of the total riboflavins, while flavinmononucleotide contributes around 10% or less and free riboflavin a few percent or less. This pattern is quiteunlike that of land plants, but rather like that in animals (Shimizu-1971). It is noteworthy that relatively highamounts of vitamin B12 are found in seaweeds (Kanazawa 1963, Hashimoto & Maeda 1953), since the B12 hadonce been thought to be distributed only in the animal kingdom.

Ito & Hori (1989)investigated the vitamin C content in seaweeds, and they reported that green seaweeds have 1-70 mg % brown seaweeds, 4-100 and red seaweeds, 1-90mg %. It is remarkable that Porphyra sp.contains a relatively high amont of vitamin C.

David (2001)studied the vitamins (A, B1, B2, B3, B12, Folic acid and C ) inGracilariaandUlva .In Ulva spvitamin A in 100g dry matter was 960 retinal U and Bcomplex (B1, B2, B3, B12 and Folic acid) was (0.06, 0.03, 8.0, 6.3 and 11.8 mg, respectively), and vitamin C (10.0 mg). InGracilariavitamin B complex (B1, B2 , B12

and Folic acid) was (0.4, 0.4, 14.4 and 2.8 mg, respectively ) vitamin C (1.1 mg).

2.3.Effect of seasons on seaweeds.

2.3.1. The seasonal effects on yield and gel properties of seaweeds.Freileet al. (1995) investigated seasonal effects on yield and gel properties ofGelidium canariensisagar for 2 intertidal populations on the northern coast of Gran Canaria.

Physical and rheological properties were measured in 1.5% w/v solutions after treatment with alkali. No significant differences were found on agar characteristics between the2 sites studied.

They were found the highest yields were obtained during summer with a maximum in June (27.8% DW) and minimum during late autumn and winter (18-18.6%).Overall quality was highest in winter (Nov.-Jan.), when gel strength peaked above 850 g/cm.

Ilyas and Sukan (1995)studied Samples ofG. verrucosa[G. gracilis] which collected monthly from the Aegean coast of Izmir Bay and analysed. They found agar yieldranged from 10.78% (October) to 30.07% (May). Gel strength, which was similar to that of technical grade agar, increased with 3,6-anhydrogalactose concentration and

decreased with reducing sugar concentration. Gelling and gel melting temperatures varied within a narrow range and were independent of season.Zinoun and Cosson (1996)studied of seasonal variation in the quality and content of iota carrageenan inC. jubatafrom the Normandy coast of France was carried out with a

view to potential commercial production. Growth increased during winter, when there was little synthesis of carrageenan and floridean starch was accumulated. Wheninorganic N content decreased, growth also decreased and stopped (May-Aug.) under high light intensity, metabolism was oriented towards synthesis of parietal carrageenans

to the detriment of reserve products such as floridean starch.Givernud et al.(1999)they studied the biology and agar composition and properties ofG. multiparita, a common species along the coasts of Morocco. They found Growth

was high in spring and autumn, and the seaweed partially decayed after fertility peaks in June and October. The agar content and composition showed seasonal variation. Agarcontent was highest in winter (30% dry weight), and decreased during growth periods to troughs in June and October (25% dry weight). Agar composition was characterized

by high 6-O-methyl galactose (38-59 mol %) and 3.6 anhydrogalactose (24-39%) contents together with galactose (12.6-25.7 mol %) an d sulfate (24-5% dry weight). Gelstrength varied between 246 and 511 g/cm2 and increased after alkali treatment to reach a maximum of 880 g/cm2.

Mouradi etal.(1999)they studied the biology and agar composition ofGelidium sesquipedalefor agar production in Morocco. The agar content varied around 40% of algaldry weight and reached a peak of 44.5% in November. Agar gel strength was highest in May and July (1000 g/cm2), and melting (90 oC) and gelling (35oC) temperatures

varied slightly.Pachecoet al.(1999)measured seasonal variation of the biomass ofG. lemaneiformisfor 18 months in Bahia de las Animas, Mexico. Showed Highest biomass per unit area(11.1 kg wet weight/m) occurred in the spring of 1995 and most of the biomass was lost by summer. Agar gel strength and yield were 891 g/cm2 and 14%, respectively for

spring samples. Biomass per unit area was also evaluated during spring from all the beds ofG. lemaneiformison the west coast of the Gulf of California (850 km). The totalbiomass estimated in 1995 was 5751404 dry t. Th e total biomass for spring of 1996 was about 30% l ess (4060246 dry t). Commercial exploitation ofG. lemaneiformis

started in the west coast of the Gulf of California in 1995.


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Benevides et al. (1999) observed remarkable variation in the haemagglutinating activity and chemical

composition of the marine red algae Gracilaria domigensis [G. domingiensis] and Gelidium pusillum during a year-

long study (February 1994 to January 1995) at a beach in Pacheco, Ceara, Brazil. Haemagglutinating activity in G.

domigensis was detected only in June while in G. pusillumit was detected from February to June. Protein levels in G.

domigensis showed small variation during the year while those in G. pusillum exhibited a pattern with maxima in

February and October. The levels of ash, carbohydrate, lipid and soluble protein showed specific variation

throughout the year for both species.

Marinho and Bourret(2003) found that the agar yield fromG. graciliswas maximum during spring (30%) and minimum during autumn (19%). InG. bursa-pastoris, theagar yield was greatest in summer (36%) and lowest in winter (23%). Agar yield from G. bursa-pastoris was positively correlated with temperature (r= 0.94P< 0.01) and

salinity (r= 0.97P< 0.01) and negatively with nitrogen content (r= -0.93P< 0.01). Agar gel strengths fluctuated from 229 to 828 g cm_2 and 23 to 168 g cm_2 for G.gracilisandG. bursa-pastoris, respectively. The gelling temperature showed significant seasonal variation for both species.

2.3.2. Seasonal effects on chemical composition of the seaweeds.

El-shazly (1956)found that seaweeds composition were changeable from time to time in the same place inAlexandria coast, also he reported that its composition were changeable from place to place in the same time.

Abdel-Fattah and Edrees (1973).Were determined seasonal changes the constituents ofUlva luctuca.Their results was showed in Table (11).

Table (11).seasonal changes the constituents ofUlva lactuca.

Date of collectionApril

1971

August

1971

November1971

Februar

y 1972Total ashTotal lipids

Unsaponified lipid fractionTotal sterols

MannitolPeriodate

ExtractionGlucuronic acid

GlucoseArabinose

XyloseRhamnose

Rh/G.A.Protein

Total amino nitrogen

24.033.070.70004

1.920.96

11.681.672.071.73

282408.702.25

20.580-400-280-15

1.651.895.732.020.670.816.081.04

33.758.13

35.893.941.570.70

1.720.612.382.450.250.391.47O-9121.164.22

24.422.911.901.12

2.010.982.51

15.741.181.581.960.12

30.136.81

Rh/G.A.-Rhamnose/glucuronic acid.

Brown et al.(1998)they studied Copper and zinc concentrations of the cosmopolitan green seaweeds Enteromorpha intestinalisandUlva lactuca were obtained for the firsttime from various sites within Otago Harbour, southern New Zealand. Spatial variation in the concentrations of both metals was found, with increasing levels at stations in the

upper harbour basin. While temporal fluctuations were apparent, a significant seasonal trend was evident only for zinc inU. lactuca concentrations varied fromapproximately 10mg g-1 dry weight in winter to 2mg g-1 in summer. Over four consecutive years of sampling there was little change in metal concentrations of either species.

Concentrations of both metals were invariably higher inE. intestinalisthanU.lactuca, at all sampling stations and at all sampling times.Fleurence et al.(1999)

Astudied the amino acid composition of the protein fraction and its changes during a sampling period from October to February were also studied.Some differences in the protein pattern shown by SDS-PAGE were found in different months, such as the presence of a 54 kDa protein in February. The protein fraction wascomposed mainly of aspartic and glutamic acids (24-35% of protein fraction, according to season) and the essential amino acids constituted 27-36% of the total fraction. Theproteins from the October sample were more sensitive to chymotrypsin than those from the February sample. For instance, t wo proteins with apparent molecular weights of

100 and 67 kDa were weakly digested by chymotrypsin in the February extract, but fully digested in the October sample. The data suggest a change in protein structuremaking it less sensitive to human intestinal juice. The glycosylation of protein extract, which was especially marked in February, could explain the differences in behaviour of

U. armoricanaproteins in response to the digestive action of enzymes. These results showed in Table (12).

Table (12). Amino acid composition ofUlva amoricanasamples collected in October 1997, December 1997 and February 1998 (g 100 g-1protein)

February 1998December 1997October 1997Essential amino acids

1.911.222.10Histidine*

3.632.312.99Soleucine6.714.645.92Leucine

1.411.722.58Methionine

6.335.077.10Phenylalanine

6.564.496.88Threoaine

5.134.055.01Valine

4.383.514.01Lysine

Non-essential amino acids

5.516.037.05Alanine

8.674.336.28Arginine

7.5411.846.09Aspartic

7.536.366.34Glycine

11.7023.3518.24Glutamic acid

5.1210.546.92Proline

1.190.471.89Hydroxyproline

6.125.605.92Serine

4.564.464.76Tyrosine

Flodin etal. (1999)studied seasonal variation in bromophenol content and bromoperoxidase activity in the green marine alga,U. lactuca. The results obtained show that bothbromophenol content and bromoperoxidase activity exhibit extreme seasonal variation, with high values in summer and low ones in winter.


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Villares et al .(2001)studied seasonal variation in the contents of different metals (Al, Cr, Cu, Fe, Mn, Ni and Zn) in two genera of macroalgae, Ulva and Enteromorpha,

studied at 22 sites on the northwest coast of Spain. The seasonal variation in the diferent metals followed a similar pattern in both seaweeds and appeared to be caused by

dilution during the period of maximum growth and concentration during periods of slow growth. Fluvial inputs of Al, Fe and Mn in autumn and winter appeared to accentuate

the latter effect the concentrations of these three metals in both macroalgae and of Cr in Enteromorphawere highest at those sites most influenced by inputs from rivers. The

background levels of Cr, Cu, Ni and Zn in the algae in summer and winter were established.

Naldi and Viaroli (2002) studied the seasonal cycle of biomass and tissue composition of Ulva rigida C. Agardh,

in relation to nitrogen availability in the water column, in 1991-1992 in the Sacca di Goro, a highly eutrophic lagoon

in the Po River Delta (Italy). Nitrate uptake rates and storage capacity were also determined in laboratory

experiments. The seasonal growth of U. rigida was related to the seasonal trend of nitrogen concentration in the

water column. U. rigida biomass increased exponentially during spring and attained peaks of about 300-400 g dry

mass (DM) m-2

in June. As biomass increased, U. rigida depleted nitrate in the water column. Thallus nitrate reserves

also declined from 100 mol N (g DM)-1

to almost undetectable levels, and total thallus nitrogen declined from 4% to

2.5% DM and 1.25% DM in 1991 and 1992, respectively. During summer, U. rigida decomposition increased, and

organic nitrogen concentrations in the water column increased.

2.4. Cell wall degradation by chemical and enzymatic treatment for improvement of protein extraction from seaweeds.

Some seaweeds belonging to the Rhodophyta and Chlorophyta contain high protein levels ( between 20 and 40%of dry weight,respectively:Fujiwara- arasakiet al. 1984)with potential uses in human and animal nutrition, suchas fish farming feeding( Fleurence, 1999). However,protein extraction from most seaweeds is difficult due to the

presence of large amounts of aninoic cell-wall polysccharides, such as the alginates of the phaeophyta or thecarrageenans of some Rhodophyta. The high content of neutral polysccharides (eg xylanes and cellulose) in somered or green seaweeds can also limit the protein accessibility. To improve the algal protein solubilization, the usein extraction buffer of additive reagents like getergents(Rice&Crowden,1987)or the application of alkalitreatment(Serotet al.,1994)is recommended in the classical extraction procedures . At the beginning of thisdecade.Amano and Noda (1990)suggested the use of algal cell wall degradation enzymes to facilitate the

extraction and the study of proteins from red seaweed.Ryoet al. (1995)Highly methylated agars wear isolated from the red seaweed,Gracilaria eucheumoides, harvested in Japan. Seaweeds were extracted exhaustively with

water at 121o using an autoclave to afford a polysaccharide from cell wall ofGracilaria eucheumoides.Chirapartet al.(1995) investigated the chemical composition of agars fromG. lemaneiformis,seaweed newly reported from Japan. They were isolated agars by a sequential

extraction of plants in water at 22 or 100C, or in boiling 20, 40 or 60% ethanol. The highest yield of agar (total carbohydrate) was obtained from the 40% ethanol extract

(55%).The highest sulfate content was obtained in non-alkali treated agars extracted with hot water (4.81%, degree of substitution (DS) 0.2). The 3, 6-anhydrogalactosecontent was highest in the 40% ethanol extract (36.1% in non-alkali treatment, 40.3% in alkali treatment). The highest methoxyl content (6.51%, DS 0.66) was obtained in the60% ethanol extract. TheG. lemaneiformisagar was composed of the biological precursor to agarobiose repeating units and agarobiose containing 6-O-methyl agarobiose and

a small amount of 2-O-methyl-alpha-L-galactopyranose residues.Fleurence et al. (1995a)studied the protein have been extracted from the edible seaweedsUlva rigidaagardh and Ulva rtundata bliding using NaOH under reductive

conditions or a two-phase system (PEG/K2CO3) produced the best protein yields. The cleavage or the limitation of the linkages between protein and polysccharides cause bythese experimental conditions prrobably explains the efficiency of these protocoles.

Fleurence et al.(1995b)studied the effect of polysaccharidases (kappa-carrageenase, beta-agarase, xylanase, cellulase) on protein extraction from 3 Rhodophyta ( Chondrus

crispus,Gracilaria verrucosa and Palmariapalmata. Kinetic parameters and optimum activity conditions for each enzyme were determined using pure substrates.. Except forP. palmata, the highest protein yields were obtained using cellulase coupled with carrageenase or agarase for an incubation period limited to 2 h. The C. crispus/cellulase +

carrageenase and G. verrucosa/cellulase + agarase systems gave 10- and 3-fold improvements, respectively, in protein extraction yield over the blank procedure. The bestoverall protein yield forP. palmatawas with xylanase only with a 14-h incubation. This study shows the possibilities of using a polysaccharidases mixture for improving

protein extractability from certain Rhodophyta.

Rebelloet al.(1996)investigated six economically important Gracilaria species, from a number of commercial sources around the world, andGracilariopsis lemaneiformis,collected from 2 Japanese localities. Agar-agar was extracted by pretreatment with various concentrations of NaOH (0, 3, 5, 7 or 10%) incubated at 80C for 2 h. In general,all species produced an agar with high gel strength after treatment with 5% NaOH, except forG. chilensisandGracilariopsis lemaneiformis, which produced agar with high

gel strength after treatment with 3, 7 or 10% NaOH.

Castro and Castro (1996)studied the optimum NaOH pretreatment strength and duration and monthly variations in gel strength, agar yield and sulfate content of agar fromG. heterocladafarmed in brackish-water canals in Leganes, Iloilo, Philippines, during the dry season (October to March), were investigated. They found The highest gelstrength (641 g/cm2) and lowest sulfate content (7.66 g/mg SO4) were obtained following pretreatment of the dried seaweed with 5% NaOH for 30 min.

Rebelloet al. (1997)investigated the chemical composition of agars extracted from economically important species of Gracilaria (G. gracilis, G. edulis, G. chilensisand G. tenuistipitata) from different geographical sources (Argentina, Brazil, Indonesia, China and Turkey) and Gracilariopsis lemaneiformiscollected from two different

Japanese localities was investigated. Agar was extracted by pretreatment with various concentrations of NaOH (3%, 5%, 7% and 10%) for 2 h at 80C. The sulfate, 3, 6-anhydrogalactose and methoxyl contents of each agar extract were analysed. High sulfate and 3,6-anhydrogalactose contents were found in non-alkali treated agar from

TurkishGracilaria gracilis(3.4%) and from ChileanG. chilensis(54.3%) after alkali treatment concentration of 5% NaOH, respectively.

Lian and Zhang (2000)investigate the effects of salinity of seawater and extraction method on agar properties of different species ofGracilaria. Five species ofGracilaria(G. lemaneiformis, G. asiatica, andG. tenuistipitatafrom wild harvest, andG. tenuistipitata var. liuiandG. sp.from culture) were collected from five regions in China in

different seasons. Salinity and temperature of seawater were recorded. Samples were pretreated with either 5% NaOH or 25% NaOH prior to agar extraction. Extracted agarsolution was filtered, gelatinized, frozen, and dehydrated. Yield, gel strength (GS), melting temperature, moisture, and sulfate content of agar were determined. They found

the yield and properties of agar varied with extraction methods, salinity, and species.G. asiaticaandG. lemaneiformistreated with 25% NaOH had higher yield (57.0% and59.3%, respectively) than with 5% NaOH (46.4% and 44.1%, respectively). GS of agar fromG. tenuistipitata var. liuiincreased with an increase in salinity in the range of

0.10 - 3.23%. The least sulfate content (0.77%) in agar was obtained from G. tenuistipitatawith the highest GS (1001g/cm2). There was no significant difference on meltingtemperatures while treated by either 25% NaOH or 5% NaOH. Sample treated by 25% NaOH had lower sulfate content as compared to 5% NaOH treatment in all cases.

2.5. Seaweeds in poultry feeding.

Abdel-salamet al. (1971),found that , egg production and utilization of feed of chicks fed on basel diet with 3% seaweeds meal (local or imported) lower than that of chicksfed on basel diet with 3% Lucerne. But they found also, no effect of addition of 3% seaweeds meal (local or imported) in the diet of laying pullets on both fertilization andhatchability ratio.

Blumet al.(1975)used different levels ofSpirulinealgae in diet hen's broiler: 0 (control) 7.5 or 15 %. Each diet was used for the feeding of 48 hybrid pullets of medium sizeduring a 24-week test period (32 to 56 weeks).they found the egg production was slightly better (47.1 g/hen/day) with 7.5 % ofSpirulines, compared to the control (45.3g/hen/day. The colour of the egg yolk was very light in the controls, but was a deep orange (above the maximum in the Roch scale) with 7.5 or 15 % of Spirulinesin the

laying hen diet.


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Incorporation of seaweeds in poultry feed was reported byChapman and Chapman (1980) and El-Deeket al. (1985).Soldevila and Almodovar (1982)reported thatsea weeds can be used as 5% of balanced diets in chickens. Stephnson (1980)found egg prouduction to increase by 20 to 30% through the addition of 8%Ascophyllumor

Laminariameal. Use of sea weeds meal up to 7% in diets of chicks and 15% for laying hens caused no problem in performance (Hand 1980 and Chapman 1980). However,higher levels reduced performance(Hoie and Sanny 1980).Jenson (1963)reported that addition of 10 to 15% brown seaweed meal to carotenoid deficient diet increased

yolk color when given to laying diets.Ben-Amotzet al.(1986)Dunaliella bardawil, a beta-carotene accumulating halotolerant algae, has been tested as a source of retinol in a chick diet. Chicks were fed diets

deficient in retinol or supplemented with retinol, synthetic beta-carotene or dry algae. After an initial lag, chicks grew equally well on diets supplemented with algae, retinolor beta-carotene. Serum and liver analyses revealed a normal content of retinol in all chicks except those grown on the retinol-deficient diet. Chicks fed the algae-

supplemented diet contained lutein but no beta-carotene in their serum, even though the ratio of beta-carotene to lutein in the algae was over 15:1. Laying hens fed with analgae-supplemented diet showed enhanced egg yolk colour attributable to a higher lutein content. No beta-carotene was present in the egg-yolk. These studies demonstrate the

possibility of using driedDunaliella bardawilas a dietary supplement which can fully satisfy the retinol requirement and also serve as a yolk-colour enhancing agent.Ross and Dominy (1990)studied the nutritional value of the blue-green algae,Spirulinaon day-old, White Leghorn cockerel chicks (120) were fed isonitrogenous diets

containing 0, 5, 10, 15, and 20% of driedSpirulina. At 3 wk of age, the growth of the chicks fed 10 and 20% ofSpirulinawas depressed, although feed efficiency was notaffected. The same other fed Hubbard male broiler chicks on experimental diets containing 0, 1.5, 3.0, 6.0, or 12.0% ofSpirulinafor 41 days. He found that the growth of the

chicks fed theSpirulinadiets was not different from that of the chicks receiving the control diet, the birds receiving the 12%Spirulinadiet grew significant slower than thechicks fed all of the otherSpirulinadiets.

Carrillo (1990)used broiler chicks from 1 day old for 8 weeks feeding on sorgam soybean diets contain 0, 5, 10, 15% seaweed (Macrocystispyrifera). There was nosignificant difference among the groups in feed intake, but body gain significantly gradually decreased by increasing in seaweeds level in the diet.

Coskun (1993)used hens 44 weeks old, in 3 groups feed for 12 weeks on abasal diet without or 0.1 or 1.0% seaweed powder. He found the feed intakeper kg egg producedwas lowest with 0.1% seaweed powder. Seaweed powder affected egg yolk colour but had no effect on feed intake.

Venkataramanet al(1994)studied the effect of sun-dried Spirulina platensis in poultry diets in a 12-weekfeeding trial by replacing either fishmeal (FM) or groundnut cake (GC) in a commercial diet with algae atisonitrogenous concentrations of 140 g/kg and 170 g/kg respectively. Additional vitamins/minerals were omittedfrom the algal diets because Spirulina is rich in them. Efficiency of food utilisation, protein efficiency ratio anddressing percentage indicated that substitution of FM or GC by alga did not affect the performance of broilers.Meat quality remained unchanged except for a more intense colour in the case of birds fed on the alga-containingdiets.

Venturaet al(1994)studied the chicken performance of seaweed (Ulva rigida) in poultry diets. The TMEn value ofU. rigida, estimated after tube-feeding, was 5.7 and 4.3MJ/kg DM for 3-week-old chickens and adult cocks (P


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65

Two algaes (Ulva and Gelidium) are collected from seven beaches to Mediterranean sea (Port Said, Balteem,

Rasheed, Abou Qeer, El-Montaza, Sidy Bishr and Kayt-Bey) for Ulva and three beaches for Gelidium (Port Said,

Balteem and Abou Qeer) and three seasons (autumn, spring and summer).

The seaweeds were washed in distillation water to removing remaining salts, sand, and epiphytes. Then it was

dried at 60C for 24h and it was milling by blender to producing analysis following:

3-2: Extraction procedures.One kg of each seaweed from each beaches were mixed together, 7 kg of Ulva and 3 kg of Gelidiuim were

crushing with grinding will, then algal powder was used for extraction trails. Each extractive procedure wasperformed with 3 replicates using (20g) algal powder obtained by crushing the dry material with a grinding mill.

3.2.1: Extraction with acetic acid. (EX1)A sub-sample (20g) of the dried algae was added to 200ml of acetic acid solution with concentration (1, 2, 3, 4

and 5% grail acetic acid in distillated water v/v) in an Evlenmeyer flask and heated for 5 h at 120C in an

autoclave then it was transferred to centrifuge (6000/min) for 10 min and it was dried for 24 h at 60C.

3.2.2: Extraction with NaOH .(EX2)A sub-sample (20g) of the dried algae was immersed in 200ml of ( 1, 2, 3, 4 and 5% NaOH in distillated waterw/v) in an Evlenmeyer flask and heated for 2h at 80C. Then it was transferred to centrifuge (6000/min) for 10min and it was dried for 24 h at 60

C.

3.2.3: Extraction with Ca(OH)2.(EX3)Each sample was ground and mixed well before use. A sub-sample (20g) of the dried algae was

immersed in 200ml of ( 1, 2, 3, 4 and 5% ) Ca(OH)2 solution in an Evlenmeyer flask and heated for 2h at 120C,then it was transferred to centrifuge (6000/min) for 10 min and it was dried for 24 h at 60

C.

3.2.4: Extraction with acetic acid and NaOH . (EX4)Each sample from (EX1) was heated with 2% NaOH solution in an Evlenmeyer flask for 2h at 80

C then dried for

24 h at 60C and crushing.

3.2.5: Extraction with polysaccharidases (EX5).

Mixed of Four types of polysaccharidases namely, cellulases, hemicelluloses, bictianase andB- glucanasewere tested.

*Enzymatic solution preparation.The cellulase, hemicelluloses, bictianase andB- glucanase powder (10 g) was added to 200 ml phosphate

buffer (0.1 M ph 6) and left under agitation in rome for at least 3 h. the solution was centrifuged at 10000 x g for20 min. the supernatant was recovered and used as the enzymatic solution. Before incubation, salts were adad (0.5M NaCL, 40 mM Mg Cl2 , 5 Mm KCl ) to the enzymatic solution as recommended by Le Gall et al. (1990) andPotin et al. (1991).

*Enzymatic extraction procedure.Enzymatic medium (10, 20, 30 or 40 ml) was added to a sub-sample (20g) of the dried algae was immersed in

100ml ( Potin et al. 1991). The suspension at ph 6 was incubated at 30C for 24 h. After this time, the algae were

filtered through a nylon mesh and dried the residue for 24 h at 60C.

3.3: The digestibility Trails:

A digestibility trial was conducted at Poultry station of Animal Production Department, Faculty of AgricultureAzhar University, Nasr City. This experiment was designed to determine the digestibility of nutrients seaweeds.

3.3.1: Birds and diets:A total of 42 Japanese Quail males (9weeks of age) were distributed at random into 7 groups in three replicates of2 birds each. Birds were housed in battery brooders. The all groups were fasted for 24 hr to empty their digestivecanal, then each groups fed one of experimental diet.A plastic tray was placed under each cage to collect excreta of both groups twice daily. The excreta were collectedquantitatively and the feed intake was measured after the 4

thday of experiment and contains during three days.

The total amount of excreta voided by each group of birds was oven-dried at 60C for 24 hr and then weighed.

Samples of feed and excreta were assayed for analysis. Water and feed were given ad libitum.

Table (13).Composition of basel diet.

Ingredients KgYellow corn,ground

59.00

Corn Gluten, 8.00


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66

60%

Soybean meal,44%

18.50

Broilerconcentrates,52%

11.00

Bone meal 1.50

Vitamins* 0.20

Minerals** 0.20NaCl 0.10

Vegetable oil 1.00

Limestone 0.50

Total 100*Etch kg contained: VA 6000000 IU, VD3 1250000 IU, VE 15000 mg, VB1 1000 mg, VB6 1000 mg, VB12 6 mg, Nicotinic acid15000 mg, Pantothenic acid 5000 mg, Biotin 50 mg, Folic acid 500 mg and choline chloride 50% 400 mg.**Etch kg contained: Mn 35 mg, Fe 40 mg, Cu 3.5 mg, Zn 25 mg, Iodine0.25 mg, Selenium 0.075 mg, Cobalt 0.10 mg, CaCO 3 1000mg.

Diets :

7experimental diets were formulating as followed,1

stdiet is practical diet (basel diet) Table (13).

2

nd

diet contents 50% diet 1 + 50% untreated Ulva.3rd

diet contents 50% diet 1 + 50% of Ulva treated with acetic acid and NaOH treatment.4

thdiet contents 50% diet 1 + 50% of Ulva treated with enzymatic treatment.

5th

diet contents 50% diet 1 + 50% of untreated Gelidium.6

thdiet contents 50% diet 1 + 50% of Gelidium treated with acetic acid and NaOH treatment.

7th

diet contents 50% diet 1 + 50% of Gelidium treated with enzymatic treatment.Chemical composition of experimental diet were shown in Table (14).Table (14).Chemical analysis of group diets were studied.

Diet

Basledi

et

55.60

3434.7

Ulvauntreated 2344.0

U

.chem 2798.0


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67

icaltreate

dU.enzymatictreated 3103.3

Gelidium

untreated 2422.3

G.chemicaltreated 2967.9

G.enzymatictre

ated 3216.9

CP= crude protein, EE= ether extract, CF= crude fiber, NFE= nitrogen free extract.


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3.3.2:Digestion coefficient determination:The apparent digestion coefficient of protein , ether extract, crude fiber and NFE were determined by the analysiseach nutrient in feed intake and faeces and then the digestibility was computed by this equation according to El-Khimsawy 2003.

Ni - Nf x 100

NiWhere Ni = nutrient in feed intake.

Nf = nutrient in faeces.

Digestion coefficition for tested algae was determination with indirect methods as following

[( Nit Nft ) ( Nip x Ncp )] x 100Nie

Where, Nit = nutrient intake in total diets.Nft = nutrient in total faeces.Nip = nutrient intake of practical diets.

Ncp = digestion coefficient for nutrient of practical diets.

Nie = nutrient intake of total algae.The nitrogen content in faces was determined following the procedure out lined byJakobsen et al, 1960as cited

byEl-Naggar (1978).

Determination of Digestible Energy:DE = GE intake GE in excreta.

Determination of gross energy :The gross energy GE of diets and faces was calculated according to West et al, 1968 and Harper et al, 1977asfollowedGE (k.cal/100g) = NFE% x 4 + EE% x 9 + CP% x 4

3.4:Chemical analysis:

Determination of major components:-Chemical analysis for moisture, crude protein, crude fiber, ether extract and ash were determined following

procedures outlined by Association of Official Analytical Chemists (A.O.A.C,1980).

3.5: Statistical analysis:Data were analyzed using general linerel model procedure (GLM) of SAS, 1993. Tow following models wereapplied:Model (1) for analysis of digestibility trail.Y ijk = + Ti+ Rj+ TRij+ e ijkWhere Y ijk is the digestibility value.T i treatments affected andi= 1,2,3.

R j seaweeds type affected andj= 1,2.TR ij interaction.eijk Residual associated withijk observation.

Model (2) for other data in present study.

Y ijk = + T i+ R j+ Sk+ TR ij+ RS jk+ TS ik+ TRS ijk+ e ijk

Where Y ijkT i treatments affected andi= 1, 2, 3, 4, 5.R j level effected and j = 1, 2, 3, 4, 5.Sk seaweeds type and k = 1, 2.TR ij, RSjk, TSik, TRS ijk interaction.eijk Residual associated withijk observation.

4. RESULTS AND DISCUSSION


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The results in the present dissertation were divided into three main parts (I) aimed to study the effect of area

and season on chemical composition foe algae (Ulva and Gelidium). (II) aimed to study Degradation of algal cell

walls to improving protein accessibility.(III) aimed to study the effect of chemical and enzymatic treatments on

digestibility of nutrients and digestible energy on Japanese quail.

4.1. Effect of area and season on chemical composition for seaweeds.4.1.1.Ulva sp.(Sea lettuce)

Moisture %Data presented in Table (15) & Fig.(5) show that regardless of season there were significant differences among

areas in moisture content in Ulva. Port Said area had significant the highest value 9.80 %, while Balteem, El-

Montazah, Rasheed and Abou-Qeer areas (8.21, 8.50, 8.52 and 8.64% respectively) had significant lowest values, with

insignificant differences among them. Sidi Bishr and Kayt-Bey had medium values.

Irrespective of area, moisture in areo-dried matter of Ulvais not significantly affected by seasons.

Season did not significantly affected moisture of Ulva in Port Said, Abou-Qeer, El-Montazah and Sidi Bishr,

but in Balteem and Kayt-Bey moisture of Ulva were significant increased in summer compared to each autumn and

spring, and in Rasheed moisture was significantly increased in summer and autumn compared to spring. The

significant lowest values of moisture during autumn were found in Balteem and during spring in Rasheed but in

summer differences among areas were closed.

Data showed in Table (15) indicated that the range of differences among experimental areas in moisture

content of Ulvawas wide (3.24%) in autumn, medium (2.27) in spring and close (1.72) in summer.

Table (15).Effect of area and season on moisture contentUlva. (As percentage to areo-dried matter).

Area Autumn Spring summer Mean

Port Said9.84

abc

0.32

10.49a

0.46

9.06abcdef

0.21

9.80a

0.27

Balteem6.87

g

0.27

8.58cdef

0.32

9.18abcdef

0.48

8.21c

0.39

Rasheed8.85

bcdef

0.54

7.77g

0.27

8.93bcdef

0.34

8.52c

0.27

Abou-Qeer9.46

abcde

0.29

8.35def

0.25

8.12efg

0.37

8.64c

0.26El-

Montaza8.29

def

0.53

8.31def

0.62

8.92bcdef

0.79

8.50c

0.34

Sidy Bishr10.11 ab

0.18

9.65 abcd

0.32

8.68 bcdef

0.18

9.48b

0.24

Kayt-Bey7.99

efg

0.86

9.00bcdef

0.37

9.84abc

0.32

8.94b

0.39

Mean8.77

A

0.28

8.88A

0.23

8.96A

0.17a,b,cmeans within column symbolized with the same letter are not significantly deferred at ( p< 0.05) level.

A,B,C. means within raw symbolized with the same letter are not significantly deferred at (p< 0.05) level.

a.b.c means within raw or column symbolized with the same letter are not significantly deferred at (p< 0.05) level.

Crude Protein %Results detected in Table (16) & Fig.(6) indicate that there were significant differences among all areas and

seasons


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Figur (5). Effect of area and season on moisture content

Ulva.

0

2

4

6

8

10

12

PortSa

id

Balte

em

Rasheed

Abou

Qeer

El-m

ontaz

a

Sidy

Bish

r

Kayt

-Bey

area

moisture%

autumnspringsummer


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64

in crude protein of Ulva. El-Montana area had significant the highest value 21.14 % followed in significantly

desending order by Rasheed (20.01%), Port Said (19.32%), Abou-Qeer and Sidi Bishr (17.35% and 17.14% with

significant difference between them), Balteem (14.7%), while Kayt-Bey had significant lowest value 11.58 %.

Summer season had significant the highest value (20.67 %) compared to each autumn and spring season (17.56 %

and 13.73 %, respectively with significant difference). The same trend of season was observed in El-Montazah,

Rasheed, Port Said, Sidi Bishr and Balteem.Table (16).Effect of area and season on crude protein content

Ulva. (As percentage to areo-dried matter).


Port Said19.57

d

0.32

15.72h

0.32

22.67b

0.24

19.32c

1.02

Balteem15.67

h

0.41

10.57j

0.41

17.87ef

0.32

14.70e

1.10

Rasheed20.87

c

0.18

16.12gh

0.33

23.03b

0.19

20.01b

1.03

Abou-Qeer16.76

gh

0.22

17.2fg

0.23

18.11ef

0.28

17.35d

0.23El-

Montaza20.91

c

0.24

15.85h

0.28

26.64a

0.31

21.14a

1.56

Sidy Bishr18.52

e

0.34

10.11j

0.44

22.8b

0.23

17.14d

1.87

Kayt-Bey 10.66j

0.6310.54 j

0.6413.55 i

0.4911.58f

0.57

Mean17.57

B

0.56

13.73C

0.66

20.67A





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Figure (6). Effect of area and season on

crude protein content Ulva.

0

5

10

15

20

2530

PortSa

id

Balte

em

Rasheed

Abo

uQeer

El-m

ontaza

Sidy

Bishr

Kayt-B

ey

area

protein%

autumnspringsummer


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66

On the other hand, Season did not significantly affected crud protein of Ulva in Abou-Qeer and Kayt-Bey with

the exception of the difference between autumn and summer in Abou-Qeer and between summer and both autumn

and spring in Kayt-Bey was significant.

Results showed in Table (16) indicate that there are similarly effects for summer and autumn on crud protein

content in Ulvaweeds in all investigated areas. This trend was not observed in spring. Moreover, Kayt-Bey recorded

significantly the lowest crud protein content during all seasons.

It is clearing that the range of differences among experimental areas in crude protein content of Ulva was wide

(13.09%) in summer, and (8.91, 7.09) in autumn and spring respectively.

Ether extract %Data presented in Table (17) & Fig.(7) showed that irrespective of seasons there was significant differences

among areas in ether extract of Ulva. Rasheed area had significant the highest value 7.53 %, while Port Said, El-

Montazah, Balteem, Abou-Qeer and Sidi Bishr areas (5.47, 5.60, 5.63, 5.64 and 6.14% respectively) had significant

lowest values, with insignificant differences among them.

Irrespective of area, ether extract in areo-dried matter of Ulvaare not significantly affected by seasons.

Season did not significantly affected ether extract of Ulvain Port Said, El-Montazah, Balteem, Abou-Qeer, Sidi

Bishr, and Kayt-Bey but in Rasheed ether extract of Ulva was significant increased in autumn compared to each

spring and summer. The significant lowest value of ether extract during autumn was found in El-Montazah but in

spring and summer differences among areas were closed.

Data showed in Table (17) illustrated that the range of differences among experimental areas in ether extract

content of Ulva was wide (4.94%) in autumn, medium in spring (2.08) and close (1.66) in summer.

Table (17).Effect of area and season on ether extracts contentUlva. (As percentage to areo-dried matter).Area Autumn Spring summer Mean

Port Said5.53

efgh

0.24

4.92gh

0.29

5.95cdefg

0.29

5.47c

0.20

Balteem5.66

defgh

0.39

5.6defgh

0.26

5.63defgh

0.36

5.63c

0.17

Rasheed9.52

a

0.29

7.19b

0.41

5.89defg

0.33

7.53a

0.56

Abou-Qeer5.62

defgh

0.40

6.31bcdef

0.15

4.98gh

0.41

5.64c

0.26

El-Montaza

4.58h

0.40

6.52bcde

0.36

5.68defgh

0.66

5.60c

0.37

Sidy Bishr6.80

bcd

0.24

5.11fgh

0.38

6.51bcde

0.41

6.14c

0.31

Kayt-Bey5.86

defg

0.26

7.14bc

0.55

6.55bcde

0.40

6.52b

0.28

Mean6.22

A

0.35

6.11A

0.22

5.88A




Crude fiber %

As shown in Table (18) & Fig.(8) there were significant effect for each areas and seasons in crude fiber of Ulva.Port Said, El-Montazah and Kayt-Bey areas had significant the highest values (27.61, 26.96 and 27.41%

respectively). While Rasheed, Abou-Qeer and Sidi Bishr areas had significant lowest values (24.78, 24.59 and

24.66%, respectively),


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Figure (7). Effect of area and season on ether

extract content Ulva.

0

2

4

6

8

10

PortSa

id

Balteem

Rasheed

AbouQ

eer

El-mon

taza

SidyB

ishr

Kayt-Be

y

area

etherextract%

autumn

spring

summer


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69

with insignificant differences among them. Balteem had medium value. Spring and summer seasons had significant

highest values (27.35 and 26.91, respectively) followed by autumn (23.62).Table (18).Effect of area and season on crude fiber content Ulva. (As percentage to areo-dried matter).


Port Said23.58

gh

0.42

25.49f

0.64

33.76b

0.64

27.61a

1.59

Balteem23.50

ghi

0.40

32.78b

0.30

20.87l

0.35

25.72b

1.81

Rasheed21.73 jkl

0.4322.97

ghijk

0.23

29.65 d0.65

24.78c1.25

Abou-Qeer22.54

hijk

0.09

22.00ijkl

0.33

29.22d

0.56

24.59c

1.18

El-Montaza21.46

kl

0.59

35.18a

0.59

24.24fg

0.71

26.96a

2.12

Sidy Bishr21.46

kl

0.53

29.52d

0.44

23.02ghij

0.46

24.66c

1.26

Kayt-Bey31.08

c

0.55

23.53ghi

0.19

27.63e

0.36

27.41a

1.12

Mean

23.62B

0.72

27.35A

1.08

26.91A


A,B,C. means within raw symbolized with the same letter are not significantly deferred at (p< 0.05) level.a.b.c means within raw or column symbolized with the same letter are not significantly deferred at (p< 0.05) level.

Season did not significantly affected crude fiber of Ulva in Rasheed and Abou-Qeer between autumn and

spring but in Balteem, El-Montazah and Sidi Bishr crude fiber of Ulvawere significant increased in spring compared

to each summer and autumn, and in Port Said crude fiber was significantly increased in summer compared to spring

in autumn compared to spring and summer.


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Figure (8). Effect of area and season on crude

fiber content Ulva.

0

5

10

15

20

25

30

35

40

PortSa

id

Balte

em

Rasheed

Abo

uQe

er

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The significant highest value of crud fiber during autumn was found in Kayt-Bey and during spring in El-Montazah

but the significant lowest value found in Port Said, in summer was found in the significant highest value in Port Said

and the significant lowest value in Balteem.

Results in Table (18) indicated that the range of differences among experimental areas in crud fiber content of

Ulvawas wide (12.89%) in summer, medium in spring and close (9.69%) in autumn.

Ash %Results in Table (19) & Fig.(9) showed that regardless of there were significant differences among areas in ash

content of Ulva. Sidi Bishr and Abou-Qeer areas had significant the highest value (13.49 and 13.22% respectively).

While Rasheed and El-Montazah, areas had significant lowest values (10.47 and 10.73%, respectively), with

insignificant differences among them. Regardless of area during autumnUlvahad significant the highest value of ash

content (14.57%) followed by spring and summer (12.05 and 9.43, respectively).

On the other hand season did not significantly affected ash of Ulva in Abou-Qeer between autumn and spring,

in Sidi Bishr between spring and summer, but in Kayt-Bey it was not significant between autumn and summer.

In Port Said, Balteem, Rasheed and El-Montazah ash of Ulvawas significant increased in autumn compared to

each spring and summer. The highest value of ash during autumn was found in Sidi Bishr and the lowest value was

found in Kayt-Bey, but during spring the significant lowest was found in Rasheed, and the lowe

tesis algas importante

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