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Phylum Porifera: The Pore Bearing Animals

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John Ela Echon
John Ela Echon

This document provides information about the phylum Porifera (sponges). It describes the general characteristics and body structure of sponges, including their three canal systems, types of spicules, and three classes of classification. The document discusses the life processes of sponges, including their filter feeding, excretion, sexual and asexual reproduction, and ecology. It analyzes a zoological investigation about collagen isolation from marine sponges and its effects on skin parameters.

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BATAAN PENINSULA STATE UNIVERSITY Telefax: (637) 2372350 Website: www.bpsu.edu.ph GRADUATE SCHOOL Email:batpenstateuyahoo.com City of Balanga 2100, Bataan, Philippines Written Report in GSE 417 Student : John Florentino E. Echon Program : Master of Arts in Education Major : General Science Course : Biology II Time : 11:30 – 3:00 Professor : LEANDRO T. OLUBIA, Ed. D. Topic : Phylum Porifera Competencies : Identify the general characteristics of organisms belonging to Phylum Porifera : Describe the body structure and canal system of sponges : Categorize sponges based on its classification : Describe the life processes of sponges : Appreciate the significant role of marine sponges : Analyse Zoological Investigation about sponges I. Introduction Sponges are the simplest form of multi-cellular animals. A sponge is a bottom-dwelling creature which attaches itself to something solid in a place where it can find enough food to grow. The scientific name for sponges is "Porifera," which translates into "pore-bearing." Etymology: From the Latin porus for pore and Ferre to bear, hence an animal with with pores. II. Body of the Report General Characteristics A sponge is a simple organism that is easy to describe. A sponge is a sedentary, filter-feeding metazoan which has a single layer of flagellated cells that drive a unidirectional current of water through its body. Such a brief description though does not do them justice. Sponges are an ancient and highly successful group of animals. In the Palaeozoic they are believed to have comprised more than half the biomass in marine reefs. They have been living in the waters of the world for more than
600 million years, and can now be found in all marine and many freshwater habitats. Some of the general characteristics of sponges are: • Simplest of all animals • Hermaphrodites • Asymmetrical • No systems for reproduction, digestion, respiration, sensory, excretion • Filter feeders • No true tissues or body systems of any type • Almost all species are sessile suspension feeders, larvae free- swimming • Multicellular • Totipotent cells: like stem cells! Body Structure:
1. Spicules are composed of calcium carbonate or silicon dioxide. Often used in sponge ID 2. Amoebocytes (archaeocyte) amoeba-like cells. It store, digest and transport food, excrete wastes, secrete skeleton give rise to buds in asexual reproduction sclerocytes - secrete spicules spongocytes - secrete spongin collencyotes - secrete collagen 3. Mesenchyme. Located beneath the pinacocytes - a gelatinous protein layer it contains the skeletal material (ie. spongin and spicules) and amoebocytes 4. Pinacocytes. It is the outer cells covering sponge; equivalent of epiderm 5. Choanocytes- similar to choanoflagellates collared cells with flagella - create water current and collect food matter 6. Porocytes. It form ostia. It is a cylindrical tube-like cells. A contractile cell regulates the diameter of the ostia. Three Canal System There are three main types of canal system in sponges. The simplest form is Asconoid, here the canals run straight through the sponge body and all the choanocytes line the central large space called the 'spongocoel'. The water enters the ostia, is drawn through to the spongocoel and leaves through a single large osculum. Asconoid sponges have cylindrical hollow bodies and tend to grow in groups attached to some object or other in relatively shallow seas. Slightly more complicated are Syconoid sponges, externally they are fairly similar to asconoid sponges except that their body wall is thicker. The canals are branched however and do not allow the water to flow straight through in to the spongocoel. Instead the water flows a twisted route through a number of canals some of which are lined with choanocytes before being expelled into the spongocoel and out through the osculum. The spongocoel is not lined with choanocytes only the canals. Syconoid sponges go through a asconoid stage in their development suggesting that they evolved from some ancestral asconoid. Syconoid sponges do not normally form groups as do asconoid sponges.
Most modern sponge species are Leuconoid. In leuconoid sponges the canal system is more complicated again with the canals being longer and more branched, they lead to special chambers whose walls are lined by choanocytes, there are no choanocytes in the canals. There is no real spongocoel just a central exit canal leading to the osculum. Leuconoid sponges tend to live in large groups with each individual sponge having its own osculum, however the borders between individual sponges are often hard to define and the sponge may act more like a large communal organism. Four General Types of Spicules Spicules are non-living aggregates of a chemical nature, secreted and made from either silica or calcium carbonate as calcite or aragonite. These spicules are important in the classification of sponges. The four main types are: 1. Monaxon- needle-like or rod-like; straight or curved 2. Tetraxon- has 4 prongs 3. Triaxon or Hexaxon- 3 or 6 rayed 4. Polyaxon- multiple short rods radiating from a common center; burr shaped, star shaped or like a child's jack. CLASSIFICATION OF PORIFERA Porifera is divided into three classes based on the skeleton they possess. 1. Calcarea (Calcispongiae) Skeleton consists mainly of calcareous spicules Examples: Leucetta, Sycon, Leucosolenia 2. Hexactinellida (Hyalospongiae). Skeleton consists mainly of siliceous spicules Examples: Euplectella, Hyalonema 3. Demospongiae. Skeleton consists mainly of spongin fibres which may be in combination with spicules Example: Spongilla
TYPICAL SPONGE FEEDING Water flowing through sponges provides food and oxygen, as well as a means for waste removal. This flow is actively generated by the beating of flagella. The water movement through some sponges is aided by ambient currents passing over raised excurrent openings. This moving water creates an area of low pressure above the excurrent openings that assists in drawing water out of the sponge. Sponges are capable of regulating the amount of flow through their bodies by the constriction of various openings. The volume of water passing through a sponge can be enormous, up to 20,000 times its volume in a single 24 hour period. In general, sponges feed by filtering bacteria from the water that passes through them. Some sponges trap roughly 90 percent of all bacteria in the water they filter. THE CARNIVOROUS EXCEPTION Sponges of the family Cladorhizidae are especially unusual in that they typically feed by capturing and digesting whole animals. That's right, these sponges are actually carnivorous! They capture small crustaceans with their spicules which act like Velcro when they come in contact with the crustacean exoskeletons. Cells then migrate around the helpless prey and digestion takes place extracellularly. EXCRETION Excretory product, mainly ammonia, is released from the body through outgoing water current. REPRODUCTION a. Sexual Reproduction Most poriferans that reproduce by sexual means are hermaphroditic and produce eggs and sperm at different times. Sperm are frequently "broadcast" into the water column. That is, sperm are created, concentrated and sent out the excurrent openings, sometimes in masses so dense that the sponges appear to be smoking. These sperm are subsequently captured by female sponges of the same species. Inside the female, the sperm are transported to eggs by special cells called archaeocytes. Fertilization occurs in the mesenchyme and the zygotes develop into ciliated larvae. Some sponges release their larvae, where others retain them for some time. Once the larvae are in the water column they settle and develop into juvenile sponges.
b. Asexual Reproduction Pieces of sponge are able to regenerate into whole new sponges. Asexual reproduction occurs by budding or by fragmentation. The buds may remain attached to the parent or separate from it, and each bud develops into a new individual. Freshwater sponges, as well as several marine species, form resistant structures called gemmules that can withstand adverse conditions such as drying or cold and later develop into new individuals. Gemmules are aggregates of sponge tissue and food, covered by a hard coating containing spicules or spongin fibers. MOVEMENT Most species are sessile as adults. Cells frequently move and rearrange themselves. Amoebocytes are highly mobile. One species, Tethya seychellensis, Red Sea, has sticky, filamentous extensions. Filaments contract and pull sponge along. ECOLOGY AND IMPORTANCE Class calcarea: Found primarily shallow water and tropical Class hexatenilleda: Marine, primarily deep water Importance: Ideal habitats for marine animals such as snails, sea stars, sea cucumbers, and shrimp. Mutually beneficial relationships with bacteria, algae and plant-like protists many are green due to these organisms living in their tissues. Deep-water sponges provide important habitat to many species of fish and invertebrates, mostly as a source of refuge from predation and adverse conditions (e.g., strong currents) and as focal sites for foraging on prey species that aggregate in sponge habitat. Others likely use sponge habitat as breeding sites. Sponges produce a wide variety of bioactive compounds which are widely used in pharmaceuticals: antibiotics, asthma, arthritis, anticancer drugs, chemicals that promote wound healing, anti- inflammatories. eg. antibiotics against bacteria such as E. coli and Staph aureus eg. Acyclovir from Caribbean sponge which is the 1st antiviral compound approved for human use fights herpes infections used since 1982
III. Review and Critiquing Zoological Investigations Title: Marine sponge collagen: isolation, characterization and effects on the skin parameters surface-pH, moisture and sebum Dieter Swatscheka,b,c, Wolfgang Schattona, Josef Kellermannd, Werner E.G. Mu¨llerb, Jo¨rg Kreuterc,* INTRODUCTION AND PROBLEMS Collagen is a fibrous protein found ubiquitously in all multicellular animals.Many pharmaceutical applications are known for collagen, e.g. shields, injectable dispersions, sponges and microparticles. Furthermore, collagen is used as a moisturiser in cosmetic cream. Moisturisers are employed for the therapy of so-called ‘dry skin’. Lipids work by the principle of occlusion, whereas humectants, such as collagen, attract water in the stratum corneum. The use of cattle as the main source for collagen has to be reconsidered because of the risks of BSE (bovine spongiform encephalopathy) and TSE (transmissible spongiform encephalopathy). One alternative is the use of porcine collagen or, much safer, collagen from sea animals, such as marine sponges. Is marine sponge can be used as alternative source of collagen? METHODS Isolation of marine sponge collagen About 50 g of ethanol-conserved sponge material were washed three times under running tap water, cut into small pieces and homogenized using a blender. 2.3. Characterization of sponge collage 2.3.1. Light microscopy Aqueous dispersions of isolated material were investigated by bright field microscopy (Standardmikroskop CH, Olympus Optical Co. GmbH, Hamburg, Germany) and by phase contrast microscopy (AH3-RFCA, Olympus Optical Co. GmbH, Hamburg, Germany). 2.3.2. Electron microscopy Twenty microliter aliquots of aqueous collagen dispersionsm were sampled for negative staining, using the singledroplet procedure. 2.4.3. In-vivo measurements Eleven female and six male volunteers from 24 to 52 years of age participated in the trials. Seventy- five mg of each preparation were applied to a 25 cm2 area of the volar forearm. The subjects were asked not to wash the forearm or to apply cosmetics to the volar forearm for a period of at least 12 h prior to the study. Results The results from the pH-measurements show that no significant or adverse changes were caused by any of these formulations. Consequently, all formulations are suitable for repeated applications, as usually recommended for the therapy of ‘dry skin’. Concerning skin- hydration both sponge collagen containing preparations as well as the commercial collagen containing preparation Shoynear Collactivew showed a slight increase. The enhancement in lipids indicated by the sebumeter represents an advantage, since the ‘dry skin’ not only lacks skin hydration but also sufficient sebum RECOMMENDATION This study clearly demonstrates that conventional collagen can be substituted by marine collagen. In addition, the currently available complicated isolation procedure can be simplified according to the method described in this publication
Title: Isolation, characterization and molecular weight determination of collagen from marine sponge Spirastrella inconstans (Dendy) Sudharsan1, P. Seedevi1, R. Saravanan2, P. Ramasamy1, S. Vasanth Kumar3, S. Vairamani1, A. Srinivasan4 and A. Shanmugam1* INTRODUCTION AND PROBLEMS Collagen is a fibrous protein found ubiquitously in all multicellular animals. Collagen is extracted mainly from the skin and bones of ruminant and porcine livestock and poultry (Vollmer and Rosenfield, 1983; Pachence, 1992; Morimura et al., 2002). However, foot-and-mouth disease, bovine spongiform encephalopathy and avian influenza have broken out frequently in recent years (Helcke, 2000; Trevitt and Singh, 2003) and hence the interest in safer sources of collagen including marine animals (fish, jellyfish, marine sponge and squid) and those living in high and cold regions without environmental pollution has greatly increased. Therefore, alternative sources of collagen should be sought. METHODS Sample collection The sponge S. inconstans was collected from Devipattinam (Lat. 9° 28' 60’ N; Long. 78° 54' 0’ E.) at 5-6 meter depth by SCUBA diving along the Southeast coast of India. Isolation of sponge collagen Collagen was isolated by following the method of Diehl-Seifer et al. (1985). Purification of crude collagen The collagen was purified with little modification of the protocol followed by Saravanan et al. (2009) through Sephadex G-50 column chromatography. Characterization of sponge collagen Total protein estimation and its homogeneity The total protein concentration was estimated by Lowry et al. (1951) method. FT-IR spectrum analysis FT-IR spectroscopy of freeze-dried sample of sponge collagen was relied on a Bio-Rad FT-IR – 40, USA. DSC measurement The shrinkage temperature (Ts) of collagen sponge matrix indicating the resistance against thermal denaturation was determined by differential scanning calorimetry (DSC 200 F3 NETZSC H). Results Total protein and collagen content In S. inconstans, the total protein content corresponds to 32% and the yield of collagen was found to be 0.16%.The sponge collagen was examined by SDS-PAGE, using a 12% resolving gel and the molecular weight of purified collagen from S. inconstans was recorded as 58 kDa (Figure 1). In the present study, the isolated crude collagen showed three bands with 80, 60 and 59 kDa molecular weight; whereas the purified collagen recorded only a single band with 58 kDa molecular weight in SDS-PAGE. RECOMMENDATION In the present study, the SEM images of sponge collagen demonstrated that all composites displayed an open and interconnected pore structure and the spicules appeared rod like structure. The chemotactic properties of collagen have many advantages in tissue engineering scaffolds (Postlethwaite et al., 1978). The sponge collgenous poriferan has provided a natural environment for cellular attachment and aggregation due to their connective tissue; it should be more complex organisms and it analogous to collagen type XIII. (no detailed recomemedation was mentioned in the study)
IV. Synthesis Sponges are the simplest form of multi-cellular animals. They are hermaphroditic, multicellular filter feeders. They have asymmetrical Body structure. It has three canal system which are the asconoid, syconoid and leuconoid. The three distinct group are the Hexactinellida (glass sponges), the Demospongia, and the Calcarea (calcareous sponges). Sponges reproduce sexually and asexually. In terms of their ecology, they are found in shallow water but primarily in deep water. Sponges provide habitat for many marine species. In terms of medical research, sponges provide a wide variety of bioactive pharmaceutical compounds, hence one of the most significant species in earth. V. References Sudharsan, S. et al (2013). Isolation, characterization and molecular weight determination of collagen from marine sponge Spirastrella inconstans (Dendy). African Journal of Biotechnology Vol. 12(5), pp. 504-511, 30 January, 2013. Retrived: August 26, 2016 Swatschek, D. et al (2001). Marine sponge collagen: isolation, characterization and effects on the skin parameters surface-pH, moisture and sebum. European Journal of Pharmaceutics and Biopharmaceutics 53 (2002) 107–113. Retrieved: August 26, 2016
VI: TASK SHEET 1. What is the significance of sponges in the ocean as filter feeders? 2. What are the three classes of the Phylum Porifera? How are they classified? 3. How do choanocytes help sponges to survive? 4. Describe the three canal system of sponges using the figure below. 5. How each canal system affects the feeding and nutrition of sponges?

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Phylum Porifera: The Pore Bearing Animals

  • 1. BATAAN PENINSULA STATE UNIVERSITY Telefax: (637) 2372350 Website: www.bpsu.edu.ph GRADUATE SCHOOL Email:batpenstateuyahoo.com City of Balanga 2100, Bataan, Philippines Written Report in GSE 417 Student : John Florentino E. Echon Program : Master of Arts in Education Major : General Science Course : Biology II Time : 11:30 – 3:00 Professor : LEANDRO T. OLUBIA, Ed. D. Topic : Phylum Porifera Competencies : Identify the general characteristics of organisms belonging to Phylum Porifera : Describe the body structure and canal system of sponges : Categorize sponges based on its classification : Describe the life processes of sponges : Appreciate the significant role of marine sponges : Analyse Zoological Investigation about sponges I. Introduction Sponges are the simplest form of multi-cellular animals. A sponge is a bottom-dwelling creature which attaches itself to something solid in a place where it can find enough food to grow. The scientific name for sponges is "Porifera," which translates into "pore-bearing." Etymology: From the Latin porus for pore and Ferre to bear, hence an animal with with pores. II. Body of the Report General Characteristics A sponge is a simple organism that is easy to describe. A sponge is a sedentary, filter-feeding metazoan which has a single layer of flagellated cells that drive a unidirectional current of water through its body. Such a brief description though does not do them justice. Sponges are an ancient and highly successful group of animals. In the Palaeozoic they are believed to have comprised more than half the biomass in marine reefs. They have been living in the waters of the world for more than
  • 2. 600 million years, and can now be found in all marine and many freshwater habitats. Some of the general characteristics of sponges are: • Simplest of all animals • Hermaphrodites • Asymmetrical • No systems for reproduction, digestion, respiration, sensory, excretion • Filter feeders • No true tissues or body systems of any type • Almost all species are sessile suspension feeders, larvae free- swimming • Multicellular • Totipotent cells: like stem cells! Body Structure:
  • 3. 1. Spicules are composed of calcium carbonate or silicon dioxide. Often used in sponge ID 2. Amoebocytes (archaeocyte) amoeba-like cells. It store, digest and transport food, excrete wastes, secrete skeleton give rise to buds in asexual reproduction sclerocytes - secrete spicules spongocytes - secrete spongin collencyotes - secrete collagen 3. Mesenchyme. Located beneath the pinacocytes - a gelatinous protein layer it contains the skeletal material (ie. spongin and spicules) and amoebocytes 4. Pinacocytes. It is the outer cells covering sponge; equivalent of epiderm 5. Choanocytes- similar to choanoflagellates collared cells with flagella - create water current and collect food matter 6. Porocytes. It form ostia. It is a cylindrical tube-like cells. A contractile cell regulates the diameter of the ostia. Three Canal System There are three main types of canal system in sponges. The simplest form is Asconoid, here the canals run straight through the sponge body and all the choanocytes line the central large space called the 'spongocoel'. The water enters the ostia, is drawn through to the spongocoel and leaves through a single large osculum. Asconoid sponges have cylindrical hollow bodies and tend to grow in groups attached to some object or other in relatively shallow seas. Slightly more complicated are Syconoid sponges, externally they are fairly similar to asconoid sponges except that their body wall is thicker. The canals are branched however and do not allow the water to flow straight through in to the spongocoel. Instead the water flows a twisted route through a number of canals some of which are lined with choanocytes before being expelled into the spongocoel and out through the osculum. The spongocoel is not lined with choanocytes only the canals. Syconoid sponges go through a asconoid stage in their development suggesting that they evolved from some ancestral asconoid. Syconoid sponges do not normally form groups as do asconoid sponges.
  • 4. Most modern sponge species are Leuconoid. In leuconoid sponges the canal system is more complicated again with the canals being longer and more branched, they lead to special chambers whose walls are lined by choanocytes, there are no choanocytes in the canals. There is no real spongocoel just a central exit canal leading to the osculum. Leuconoid sponges tend to live in large groups with each individual sponge having its own osculum, however the borders between individual sponges are often hard to define and the sponge may act more like a large communal organism. Four General Types of Spicules Spicules are non-living aggregates of a chemical nature, secreted and made from either silica or calcium carbonate as calcite or aragonite. These spicules are important in the classification of sponges. The four main types are: 1. Monaxon- needle-like or rod-like; straight or curved 2. Tetraxon- has 4 prongs 3. Triaxon or Hexaxon- 3 or 6 rayed 4. Polyaxon- multiple short rods radiating from a common center; burr shaped, star shaped or like a child's jack. CLASSIFICATION OF PORIFERA Porifera is divided into three classes based on the skeleton they possess. 1. Calcarea (Calcispongiae) Skeleton consists mainly of calcareous spicules Examples: Leucetta, Sycon, Leucosolenia 2. Hexactinellida (Hyalospongiae). Skeleton consists mainly of siliceous spicules Examples: Euplectella, Hyalonema 3. Demospongiae. Skeleton consists mainly of spongin fibres which may be in combination with spicules Example: Spongilla
  • 5. TYPICAL SPONGE FEEDING Water flowing through sponges provides food and oxygen, as well as a means for waste removal. This flow is actively generated by the beating of flagella. The water movement through some sponges is aided by ambient currents passing over raised excurrent openings. This moving water creates an area of low pressure above the excurrent openings that assists in drawing water out of the sponge. Sponges are capable of regulating the amount of flow through their bodies by the constriction of various openings. The volume of water passing through a sponge can be enormous, up to 20,000 times its volume in a single 24 hour period. In general, sponges feed by filtering bacteria from the water that passes through them. Some sponges trap roughly 90 percent of all bacteria in the water they filter. THE CARNIVOROUS EXCEPTION Sponges of the family Cladorhizidae are especially unusual in that they typically feed by capturing and digesting whole animals. That's right, these sponges are actually carnivorous! They capture small crustaceans with their spicules which act like Velcro when they come in contact with the crustacean exoskeletons. Cells then migrate around the helpless prey and digestion takes place extracellularly. EXCRETION Excretory product, mainly ammonia, is released from the body through outgoing water current. REPRODUCTION a. Sexual Reproduction Most poriferans that reproduce by sexual means are hermaphroditic and produce eggs and sperm at different times. Sperm are frequently "broadcast" into the water column. That is, sperm are created, concentrated and sent out the excurrent openings, sometimes in masses so dense that the sponges appear to be smoking. These sperm are subsequently captured by female sponges of the same species. Inside the female, the sperm are transported to eggs by special cells called archaeocytes. Fertilization occurs in the mesenchyme and the zygotes develop into ciliated larvae. Some sponges release their larvae, where others retain them for some time. Once the larvae are in the water column they settle and develop into juvenile sponges.
  • 6. b. Asexual Reproduction Pieces of sponge are able to regenerate into whole new sponges. Asexual reproduction occurs by budding or by fragmentation. The buds may remain attached to the parent or separate from it, and each bud develops into a new individual. Freshwater sponges, as well as several marine species, form resistant structures called gemmules that can withstand adverse conditions such as drying or cold and later develop into new individuals. Gemmules are aggregates of sponge tissue and food, covered by a hard coating containing spicules or spongin fibers. MOVEMENT Most species are sessile as adults. Cells frequently move and rearrange themselves. Amoebocytes are highly mobile. One species, Tethya seychellensis, Red Sea, has sticky, filamentous extensions. Filaments contract and pull sponge along. ECOLOGY AND IMPORTANCE Class calcarea: Found primarily shallow water and tropical Class hexatenilleda: Marine, primarily deep water Importance: Ideal habitats for marine animals such as snails, sea stars, sea cucumbers, and shrimp. Mutually beneficial relationships with bacteria, algae and plant-like protists many are green due to these organisms living in their tissues. Deep-water sponges provide important habitat to many species of fish and invertebrates, mostly as a source of refuge from predation and adverse conditions (e.g., strong currents) and as focal sites for foraging on prey species that aggregate in sponge habitat. Others likely use sponge habitat as breeding sites. Sponges produce a wide variety of bioactive compounds which are widely used in pharmaceuticals: antibiotics, asthma, arthritis, anticancer drugs, chemicals that promote wound healing, anti- inflammatories. eg. antibiotics against bacteria such as E. coli and Staph aureus eg. Acyclovir from Caribbean sponge which is the 1st antiviral compound approved for human use fights herpes infections used since 1982
  • 7. III. Review and Critiquing Zoological Investigations Title: Marine sponge collagen: isolation, characterization and effects on the skin parameters surface-pH, moisture and sebum Dieter Swatscheka,b,c, Wolfgang Schattona, Josef Kellermannd, Werner E.G. Mu¨llerb, Jo¨rg Kreuterc,* INTRODUCTION AND PROBLEMS Collagen is a fibrous protein found ubiquitously in all multicellular animals.Many pharmaceutical applications are known for collagen, e.g. shields, injectable dispersions, sponges and microparticles. Furthermore, collagen is used as a moisturiser in cosmetic cream. Moisturisers are employed for the therapy of so-called ‘dry skin’. Lipids work by the principle of occlusion, whereas humectants, such as collagen, attract water in the stratum corneum. The use of cattle as the main source for collagen has to be reconsidered because of the risks of BSE (bovine spongiform encephalopathy) and TSE (transmissible spongiform encephalopathy). One alternative is the use of porcine collagen or, much safer, collagen from sea animals, such as marine sponges. Is marine sponge can be used as alternative source of collagen? METHODS Isolation of marine sponge collagen About 50 g of ethanol-conserved sponge material were washed three times under running tap water, cut into small pieces and homogenized using a blender. 2.3. Characterization of sponge collage 2.3.1. Light microscopy Aqueous dispersions of isolated material were investigated by bright field microscopy (Standardmikroskop CH, Olympus Optical Co. GmbH, Hamburg, Germany) and by phase contrast microscopy (AH3-RFCA, Olympus Optical Co. GmbH, Hamburg, Germany). 2.3.2. Electron microscopy Twenty microliter aliquots of aqueous collagen dispersionsm were sampled for negative staining, using the singledroplet procedure. 2.4.3. In-vivo measurements Eleven female and six male volunteers from 24 to 52 years of age participated in the trials. Seventy- five mg of each preparation were applied to a 25 cm2 area of the volar forearm. The subjects were asked not to wash the forearm or to apply cosmetics to the volar forearm for a period of at least 12 h prior to the study. Results The results from the pH-measurements show that no significant or adverse changes were caused by any of these formulations. Consequently, all formulations are suitable for repeated applications, as usually recommended for the therapy of ‘dry skin’. Concerning skin- hydration both sponge collagen containing preparations as well as the commercial collagen containing preparation Shoynear Collactivew showed a slight increase. The enhancement in lipids indicated by the sebumeter represents an advantage, since the ‘dry skin’ not only lacks skin hydration but also sufficient sebum RECOMMENDATION This study clearly demonstrates that conventional collagen can be substituted by marine collagen. In addition, the currently available complicated isolation procedure can be simplified according to the method described in this publication
  • 8. Title: Isolation, characterization and molecular weight determination of collagen from marine sponge Spirastrella inconstans (Dendy) Sudharsan1, P. Seedevi1, R. Saravanan2, P. Ramasamy1, S. Vasanth Kumar3, S. Vairamani1, A. Srinivasan4 and A. Shanmugam1* INTRODUCTION AND PROBLEMS Collagen is a fibrous protein found ubiquitously in all multicellular animals. Collagen is extracted mainly from the skin and bones of ruminant and porcine livestock and poultry (Vollmer and Rosenfield, 1983; Pachence, 1992; Morimura et al., 2002). However, foot-and-mouth disease, bovine spongiform encephalopathy and avian influenza have broken out frequently in recent years (Helcke, 2000; Trevitt and Singh, 2003) and hence the interest in safer sources of collagen including marine animals (fish, jellyfish, marine sponge and squid) and those living in high and cold regions without environmental pollution has greatly increased. Therefore, alternative sources of collagen should be sought. METHODS Sample collection The sponge S. inconstans was collected from Devipattinam (Lat. 9° 28' 60’ N; Long. 78° 54' 0’ E.) at 5-6 meter depth by SCUBA diving along the Southeast coast of India. Isolation of sponge collagen Collagen was isolated by following the method of Diehl-Seifer et al. (1985). Purification of crude collagen The collagen was purified with little modification of the protocol followed by Saravanan et al. (2009) through Sephadex G-50 column chromatography. Characterization of sponge collagen Total protein estimation and its homogeneity The total protein concentration was estimated by Lowry et al. (1951) method. FT-IR spectrum analysis FT-IR spectroscopy of freeze-dried sample of sponge collagen was relied on a Bio-Rad FT-IR – 40, USA. DSC measurement The shrinkage temperature (Ts) of collagen sponge matrix indicating the resistance against thermal denaturation was determined by differential scanning calorimetry (DSC 200 F3 NETZSC H). Results Total protein and collagen content In S. inconstans, the total protein content corresponds to 32% and the yield of collagen was found to be 0.16%.The sponge collagen was examined by SDS-PAGE, using a 12% resolving gel and the molecular weight of purified collagen from S. inconstans was recorded as 58 kDa (Figure 1). In the present study, the isolated crude collagen showed three bands with 80, 60 and 59 kDa molecular weight; whereas the purified collagen recorded only a single band with 58 kDa molecular weight in SDS-PAGE. RECOMMENDATION In the present study, the SEM images of sponge collagen demonstrated that all composites displayed an open and interconnected pore structure and the spicules appeared rod like structure. The chemotactic properties of collagen have many advantages in tissue engineering scaffolds (Postlethwaite et al., 1978). The sponge collgenous poriferan has provided a natural environment for cellular attachment and aggregation due to their connective tissue; it should be more complex organisms and it analogous to collagen type XIII. (no detailed recomemedation was mentioned in the study)
  • 9. IV. Synthesis Sponges are the simplest form of multi-cellular animals. They are hermaphroditic, multicellular filter feeders. They have asymmetrical Body structure. It has three canal system which are the asconoid, syconoid and leuconoid. The three distinct group are the Hexactinellida (glass sponges), the Demospongia, and the Calcarea (calcareous sponges). Sponges reproduce sexually and asexually. In terms of their ecology, they are found in shallow water but primarily in deep water. Sponges provide habitat for many marine species. In terms of medical research, sponges provide a wide variety of bioactive pharmaceutical compounds, hence one of the most significant species in earth. V. References Sudharsan, S. et al (2013). Isolation, characterization and molecular weight determination of collagen from marine sponge Spirastrella inconstans (Dendy). African Journal of Biotechnology Vol. 12(5), pp. 504-511, 30 January, 2013. Retrived: August 26, 2016 Swatschek, D. et al (2001). Marine sponge collagen: isolation, characterization and effects on the skin parameters surface-pH, moisture and sebum. European Journal of Pharmaceutics and Biopharmaceutics 53 (2002) 107–113. Retrieved: August 26, 2016
  • 10. VI: TASK SHEET 1. What is the significance of sponges in the ocean as filter feeders? 2. What are the three classes of the Phylum Porifera? How are they classified? 3. How do choanocytes help sponges to survive? 4. Describe the three canal system of sponges using the figure below. 5. How each canal system affects the feeding and nutrition of sponges?