A histochemical and morphological study of the mucus producing pedal gland system in Latia neritoides (Mollusca; Gastropoda; Hygrophila).

The freshwater gastropod Latia neritoides is endemic to the streams of New Zealand's North Island. This species has evolved a unique defence system: it exudes a luminescent mucus thought to deter predators. While the bioluminescence itself has been investigated before, the underlying gland system has remained unstudied and relevant information to understand the defence system has been missing till now. For the release of the glowing mucus of L. neritoides two places of origin were assumed: the lateral foot area or the mantel cavity. In this study the focus was on the first suggestion. To gain insight into the defence system, morphological as well as histochemical analyses were performed involving all secretory gland types in the sub-epithelial foot layer. The results were compared with the foot gland system of Neritina sp., a snail living in a comparable habitat, but using a different survival strategy. The gland types of the two gastropods were compared and their mucus types were investigated. Seven subepithelial gland cell types can be distinguished in the foot region of L. neritoides. Neritina sp., in contrast, has six gland cell types of which three laterally located ones are epithelial. Both species show a pedal gland in the anterior foot region. A striking difference between the species are two prominent subepithelial gland cell types (L1l/L2l) in the lateral foot area of L. neritoides, which are missing in Neritina sp. These gland cells are distributed throughout the entire lateral foot area of L. neritoides and make up about 85% of the mucus gland cells in this area. Defence mucus and trail mucus of L. neritoides show different specificities in lectin staining, but are not equally represented in the gland cell types. Yet, based on the huge size and high density of L1l and L2L, we envision a role for these gland types in the defence system.

Polymer Based Bioadhesive Biomaterials for Medical Application-A Perspective of Redefining Healthcare System Management.

This article deliberates about the importance of polymer-based bioadhesive biomaterials' medical application in healthcare and in redefining healthcare management. Nowadays, the application of bioadhesion in the health sector is one of the great interests for various researchers, due to recent advances in their formulation development. Actually, this area of study is considered as an active multidisciplinary research approach, where engineers, scientists (including chemists, physicists, biologists, and medical experts), material producers and manufacturers combine their knowledge in order to provide better healthcare. Moreover, while discussing the implications of value-based healthcare, it is necessary to mention that health comprises three main domains, namely, physical, mental, and social health, which not only prioritize the quality healthcare, but also enable us to measure the outcomes of medical interventions. In addition, this conceptual article provides an understanding of the consequences of the natural or synthetic polymer-based bioadhesion of biomaterials, and its significance for redefining healthcare management as a novel approach. Furthermore, the research assumptions highlight that the quality healthcare concept has recently become a burning topic, wherein healthcare service providers, private research institutes, government authorities, public service boards, associations and academics have taken the initiative to restructure the healthcare system to create value for patients and increase their satisfaction, and lead ultimately to a healthier society.

Characterization of the Arion vulgaris pedal gland system.

The most common European gastropod species, Arion vulgaris, is one of the most troublesome pests for private garden owners and commercial agriculturists. The sticky and hard to remove secretion produced by these animals allows them to overcome most artificial and natural barriers. However, this highly adherent biopolymer has recently shown great potential for novel wound-healing applications in medicine. Nevertheless, our knowledge of the underlying gland system is still limited and few studies on the ventral gland system are available. We studied the lateral and ventral pedal glands in Arion vulgaris to determine their secretory content histochemically and through lectin assays. Using these histological and histochemical methods we differentiate five gland types with different mucus composition in the lateral pedal region of the foot of Arion vulgaris. These contain sulphated and carboxylated mucosubstances (positive Alcian blue staining) but lack hexose-containing mucosubstances (negative PAS staining). In the ventral pedal region, four gland types can be differentiated producing sulphated and carboxylated mucosubstances. Within the ventral mucus, a high affinity for the lectins PNA and WGA is observed. While the lateral glands are histochemically negative for PAS, a positive staining with the lectin JAC is observed. Arion vulgaris shows clear morphological differences from other arionid species. This raises the question whether the variation in the chemistry of the secretory material and mucus composition is the result of different functions and/or is related to the animals' different environmental conditions. A comparison of some glands of Arion vulgaris with those of the helicid species Helix pomatia and Cepaea hortensis indicates morphological similarities.

Current State of Bone Adhesives-Necessities and Hurdles.

The vision of gluing two bone fragments with biodegradable and biocompatible adhesives remains highly fascinating and attractive to orthopedic surgeons. Possibly shorter operation times, better stabilization, lower infection rates, and unnecessary removal make this approach very appealing. After 30 years of research in this field, the first adhesive systems are now appearing in scientific reports that may fulfill the comprehensive requirements of bioadhesives for bone. For a successful introduction into clinical application, special requirements of the musculoskeletal system, challenges in the production of a bone adhesive, as well as regulatory hurdles still need to be overcome. In this article, we will give an overview of existing synthetic polymers, biomimetic, and bio-based adhesive approaches, review the regulatory hurdles they face, and discuss perspectives of how bone adhesives could be efficiently introduced into clinical application, including legal regulations.

Biomechanical properties of fishing lines of the glowworm Arachnocampa luminosa (Diptera; Keroplatidae).

Animals use adhesive secretions in highly diverse ways, such as for settlement, egg anchorage, mating, active or passive defence, etc. One of the most interesting functions is the use of bioadhesives to capture prey, as the bonding has to be performed within milliseconds and often under unfavourable conditions. While much is understood about the adhesive and biomechanical properties of the threads of other hunters such as spiders, barely anything is documented about those of the New Zealand glowworm Arachnocampa luminosa. We analysed tensile properties of the fishing lines of the New Zealand glowworm Arachnocampa luminosa under natural and dry conditions and measured their adhesion energy to different surfaces. The capture system of A. luminosa is highly adapted to the prevailing conditions (13-15 °C, relative humidity of 98%) whereby the wet fishing lines only show a bonding ability at high relative humidity (>80%) with a mean adhesive energy from 20-45 N/m and a stronger adhesion to polar surfaces. Wet threads show a slightly higher breaking strain value than dried threads, whereas the tensile strength of wet threads was much lower. The analyses show that breaking stress and strain values in Arachnocampa luminosa were very low in comparison to related Arachnocampa species and spider silk threads but exhibit much higher adhesion energy values. While the mechanical differences between the threads of various Arachnocampa species might be consequence of the different sampling and handling of the threads prior to the tests, differences to spiders could be explained by habitat differences and differences in the material ultrastructure. Orb web spiders produce viscid silk consisting of ß-pleated sheets, whereas Arachnocampa has cross-ß-sheet crystallites within its silk. As a functional explanation, the low tear strength for A. luminosa comprises a safety mechanism and ensures the entire nest is not pulled down by prey which is too heavy.

The structure of the cutaneous pedal glands in the banded snail Cepaea hortensis (Müller, 1774).

Although gastropods have been crawling through the ocean and on the land for 60 million years, we still know very little about the sticky mucus produced in their foot. Most research has been focused on marine species in particular and, to a lesser extent, on the well-known terrestrial species Arion vulgaris and Cornu aspersum. Within this study, we aim to characterize the foot anatomy of a smaller representative of the family Helicidae, the banded snail Cepaea hortensis. We are particularly interested in the microanatomy of the foot glands, their position, and the histochemical nature of their secretory content. Characterization of the dorsal foot region of Cepaea hortensis reveals four glands, differing in their size and in the granules produced. Histochemically, three of them react positively for sugars (PAS staining and lectin affinity tests for mannose, glucose and N-acetyl-d-glucosamine) and acidic proteins (positive Alcian blue and Toluidine blue staining), indicating the presence of acidic glycosaminoglycans. The fourth gland type does not react to any of these dyes. The ventral pedal region includes two different gland types, which are positive for the presence of acidic glycoproteins, with a lectin affinity for mannose only. A comparison with Helix pomatia indicates differences regarding the number of glands and their contents. In Helix, only three gland types are described in the dorsal region of the foot, which show a similar granular appearance but nevertheless differ in their chemical composition. Congruently, there are two gland types in the ventral region in both species, whereas in Helix an additional sugar moiety is found. This raises the question whether these differences between the pedal glandular systems of both helicid species are the result of protection or size-related adaptations, as they occur in the same habitat.

Chemical characterization of the adhesive secretions of the salamander Plethodon shermani (Caudata, Plethodontidae).

Salamanders have developed a wide variety of antipredator mechanisms, including tail autotomy, colour patterns, and noxious skin secretions. As an addition to these tactics, the red-legged salamander (Plethodon shermani) uses adhesive secretions as part of its defensive strategy. The high bonding strength, the fast-curing nature, and the composition of the biobased materials makes salamander adhesives interesting for practical applications in the medical sector. To understand the adhesive secretions of P. shermani, its components were chemically analysed by energy dispersive X-ray spectroscopy (EDX), inductively coupled plasma mass spectrometry (ICP-MS), amino acid analysis, and spectroscopy (ATR-IR, Raman). In addition, proteins were separated by gel-electrophoresis and selected spots were characterised by peptide mass fingerprinting. The salamander secretion contains a high amount of water and predominantly proteins (around 77% in the dry stage). The gel-electrophoresis and peptide mass fingerprint analyses revealed a de novo set of peptides/proteins, largely with a pI between 5.0 and 8.0 and a molecular mass distribution between 10 and 170 kDa. Only low homologies with other proteins present in known databases could be identified. The results indicate that the secretions of the salamander Plethodon clearly differ chemically from those shown for other glue-producing terrestrial or marine species and thus represent a unique glue system.

Salamanders on the bench - A biocompatibility study of salamander skin secretions in cell cultures.

Salamanders have evolved a wide variety of antipredator mechanisms and behavior patterns, including toxins and noxious or adhesive skin secretions. The high bonding strength of the natural bioadhesives makes these substances interesting for biomimetic research and applications in industrial and medical sectors. Secretions of toxic species may help to understand the direct effect of harmful substances on the cellular level. In the present study, the biocompatibility of adhesive secretions from four salamander species (Plethodon shermani, Plethodon glutinosus, Ambystoma maculatum, Ambystoma opacum) were analyzed using the MTT assay in cell culture and evaluated against toxic secretions of Pleurodeles waltl, Triturus carnifex, Pseudotriton ruber, Tylototriton verrucosus, and Salamandra salamandra. Their effect on cells was tested in direct contact (direct culture) or under the influence of the extract (indirect exposure) in accordance with the protocol of the international standard norm ISO 10993-5. Human dermal fibroblasts (NHDF), umbilical vein endothelial cells (HUVEC), and articular chondrocytes (HAC), as well as the cell lines C2C12 and L929 were used in both culture types. While the adhesive secretions from Plethodon shermani are cytocompatible and those of Ambystoma opacum are even advantageous, those of Plethodon glutinosus and Ambystoma maculatum appear to be cytotoxic to NDHF and HUVEC. Toxic secretions from Salamandra salamandra exhibited harmful effects on all cell types. Pseudotriton ruber and Triturus carnifex secretions affected certain cell types marginally; those from Pleurodeles waltl and Tylototriton verrucosus were generally well tolerated. The study shows for the first time the effect of salamander secretions on the viability of different cell types in culture. Two adhesive secretions appeared to be cell compatible and are therefore promising candidates for future investigations in the field of medical bioadhesives. Among the toxic secretions tested, only two of the five had a harmful effect on cells, indicating different cell toxicity mechanisms.

Snail mucus - glandular origin and composition in Helix pomatia.

Apart from their well-known culinary use, gastropod species such as Helix, which have a hydrogel-like mucus, are increasingly being exploited for cosmetic, bioengineering and medical applications. However, not only are the origin and composition of these "sticky" secretions far from being fully characterized, the number and morphology of the mucus glands involved is also uncertain. This study aims to characterize in detail the cutaneous glands of the Helix pomatia foot on morphological, histochemical and immunohistochemical levels. Hereby the focus is on the gland position and appearance on the foot sole as well as on the chemical nature of the different gland secretions. At least five different gland types can be distinguished by their microanatomy; three are located on the dorsal side and two on the ventral side of the foot sole. Most glands are reactive for acidic proteins and sugars such as mannose and fucose, indicating the presence of acidic glycosaminoglycans. One dorsal gland type shows high reactivity for acidic proteins only. The isolated mucus includes a certain amount of the elements chlorine, potassium and calcium; evidence for lipids was also confirmed in the isolated mucus. The present results for Helix pomatia show a clear difference in the number of glands compared to the related species Helix aspersa (only four mucus glands); histochemically, the glands of both species similarly produce acidic proteins as well as acidic glycosaminoglycans. While calcium ions are known to play a role in mucus formation, the presence and function of other ions such as potassium still need to be clarified.

Characterization of the adhesive dermal secretion of Euprymna scolopes Berry, 1913 (Cephalopoda).

Bio-adhesion is a common and crucial process in nature and is used by several different species for camouflage, prey capture, hatching or to avoid drifting. Four genera of cephalopods belonging to four different families (Euprymna, Sepiolidae; Idiosepius, Idiosepiidae; Nautilus, Nautilidae; and Sepia, Sepiidae) produce glue for temporary attachment. Euprymna species live in near-shore benthic habitats of the Indo-Pacific Ocean, are nocturnal and bury into the seafloor during the day. The animals secrete adhesives through their epithelial glands to completely coat themselves with sand. In cases of danger, they instantaneously release the sandy coat as a sinking decoy to deflect predators. Earlier morphological investigations have shown that the adhesive gland cells of Euprymna scolopes are scattered on the dorsal epidermis. It has been proposed that neutral mucopolysaccharides, secreted by one gland type (goblet cells), are responsible for adhesion, whereas the release of the glue could be caused by acidic mucoproteins produced by ovate cells in the ventral epidermis. The ultrastructural re-investigation of the Euprymna epithelium in this study has indicated the presence of a new gland type (named flask cell), exclusively located in the dorsal epithelium and always neighboured to the known goblet cells. Based on our histochemical observations, the secretory material of the ovate cells does not display a strong reaction to tests for acidic groups, as had been previously assumed. Within the dermis, a large muscle network was found that was clearly distinctive from the normal mantle musculature. Based on our data, an antagonistic gland system, as previously proposed, seems to be unlikely for Euprymna scolopes. We hypothesize that the adhesive secretion is formed by two gland types (goblet and flask cells). The release of the sand coat may occur mechanically, i.e. by contraction of the dermal mantle muscle, and not chemically through the ovate cells.

Characterization of the Fishing Lines in Titiwai (=Arachnocampa luminosa Skuse, 1890) from New Zealand and Australia.

Animals use adhesive secretions in a plethora of ways, either for attachment, egg anchorage, mating or as either active or passive defence. The most interesting function, however, is the use of adhesive threads to capture prey, as the bonding must be performed within milliseconds and under unsuitable conditions (movement of prey, variable environmental conditions, unfavourable attack angle, etc.) to be nonetheless successful. In the following study a detailed characterization of the prey capture system of the world-renowned glowworm group Arachnocampa from the macroscopic to the ultrastructural level is performed. The data reveal that the adhesive droplets consist mostly of water and display hygroscopic properties at varying humidity levels. The droplet core of Arachnocampa luminosa includes a certain amount of the elements sodium, sulphur and potassium (beside carbon, oxygen and nitrogen), while a different element composition is found in the two related species A. richardsae and A. tasmaniensis. Evidence for lipids, carbohydrates and proteins was negative on the histochemical level, however X-ray photoelectron spectroscopy confirm the presence of peptides within the droplet content. Different to earlier assumptions, the present study indicates that rather than oxalic acid, urea or uric acid are present in the adhesive droplets, presumably originating from the gut. Comparing the capture system in Arachnocampa with those of orb-spiders, large differences appear not only regarding the silky threads, but also, in the composition, hygroscopic properties and size of the mucous droplets.

Morphological characterization of the glandular system in the salamander Plethodon shermani (Caudata, Plethodontidae).

Amphibians have evolved a wide variety of mechanisms that provide a certain degree of protection against predators, including camouflage, tail autonomy, encounter behavior and noxious or toxic skin secretions. In addition to these strategies, some amphibians release a glue-like secretion onto the surface of their skin when threatened. While some information regarding the origin and production of these adhesive secretions is available for frogs such as Notaden bennetti, these aspects are only partially understood in salamanders. We contribute to an earlier study and provide additional information regarding the origin, production, and characterization of the adhesive secretion in the red-legged salamander (Plethodon shermani) at a microanatomical level. When stressed, this salamander secretes a milky, viscous liquid from its dorsal and ventral skin. This secretion is extremely adhesive and hardens within seconds upon exposure to air. This study describes two cutaneous gland types (mucous and granular) in the dorsal and ventral epithelial tissue that differ considerably in their secretory content. While the smaller mucous glands contains flocculent to granular material, mostly acidic glycoproteins, the granular glands synthesize various granules of differing size and density that consist of basic proteinaceous material. The results strongly indicate that the secretions of both gland types from the dorsal as well as the ventral side form the adhesive mucus in Plethodon shermani, consisting of basic and acidic glycoproteins, glycoconjugates with mannose and α-L-fucose residues as well as lipid components.

The chemistry of stalked barnacle adhesive (Lepas anatifera).

The results of the first chemical analysis of the adhesive of Lepas anatifera, a stalked barnacle, are presented. A variety of elements were identified in scanning electron microscopy with energy dispersive spectrometry (SEM-EDS) of the adhesive, including Na, Mg, Ca, Cl, S, Al, Si, K and Fe; however, protein-metal interactions were not detected in Raman spectra of the adhesive. Elemental signatures from SEM-EDS of L. anatifera adhesive glands were less varied. Phosphorous was mostly absent in adhesive samples; supporting previous studies showing that phosphoserines do not play a significant role in adult barnacle adhesion. Disulfide bridges arising from Cys dimers were also investigated; Raman analysis showed weak evidence for S-S bonds in L. anatifera. In addition, there was no calcium carbonate signal in the attenuated total reflectance Fourier transform infrared spectra of L. anatifera adhesive, unlike several previous studies in other barnacle species. Significant differences were observed between the Raman spectra of L. anatifera and Balanus crenatus; these and a range of Raman peaks in the L. anatifera adhesive are discussed. Polysaccharide was detected in L. anatifera adhesive but the significance of this awaits further experiments. The results demonstrate some of the diversity within barnacle species in the chemistry of their adhesives.

Alterations in the mantle epithelium during transition from hatching gland to adhesive organ of Idiosepius pygmaeus (Mollusca, Cephalopoda).

Epithelial gland systems play an important role in marine molluscs in fabricating lubricants, repellents, fragrances, adhesives or enzymes. In cephalopods the typically single layered epithelium provides a highly dynamic variability and affords a rapid rebuilding of gland cells. While the digestive hatching gland (also named Hoyle organ) is obligatory for most cephalopods, only four genera (Nautilus, Sepia, Euprymna and Idiosepius) produce adhesive secretions by means of glandular cells in an adhesive area on the mantle or tentacles. In Idiosepius this adhesive organ is restricted to the posterior part of the fin region on the dorsal mantle side and well developed in the adult stage. Two gland cell types could be distinguished, which produce different contents of the adhesive. During the embryonic development the same body area is occupied by the temporary hatching gland. The question arises, in which way the hatching gland degrades and is replaced by the adhesive gland. Ultrastructural analyses as well as computer tomography scans were performed to monitor the successive post hatching transformation in the mantle epithelium from hatching gland degradation to the formation of the adhesive organ. According to our investigations the hatching gland cells degrade within about 1 day after hatching by a type of programmed cell death and leave behind a temporary cellular gap in this area. First glandular cells of the adhesive gland arise 7 days after hatching and proceed evenly over the posterior mantle epithelium. In contrast, the accompanying reduction of a part of the dorsal mantle musculature is already established before hatching. The results demonstrate a distinct independence between the two gland systems and illustrate the early development of the adhesive organ as well as the corresponding modifications within the mantle.

Biochemical analyses of the cement float of the goose barnacle Dosima fascicularis--a preliminary study.

The goose barnacle Dosima fascicularis produces an excessive amount of adhesive (cement), which has a double function, being used for attachment to various substrata and also as a float (buoy). This paper focuses on the chemical composition of the cement, which has a water content of 92%. Scanning electron microscopy with EDX was used to measure the organic elements C, O and N in the foam-like cement. Vibrational spectroscopy (FTIR, Raman) provided further information about the overall secondary structure, which tended towards a ß-sheet. Disulphide bonds could not be detected by Raman spectroscopy. The cystine, methionine, histidine and tryptophan contents were each below 1% in the cement. Analyses of the cement revealed a protein content of 84% and a total carbohydrate content of 1.5% in the dry cement. The amino acid composition, 1D/2D-PAGE and MS/MS sequence analysis revealed a de novo set of peptides/proteins with low homologies with other proteins such as the barnacle cement proteins, largely with an acidic pI between 3.5 and 6.0. The biochemical composition of the cement of D. fascicularis is similar to that of other barnacles, but it shows interesting variations.

Morphology of the cement apparatus and the cement of the buoy barnacle Dosima fascicularis (Crustacea, Cirripedia, Thoracica, Lepadidae).

Barnacles produce a proteinaceous adhesive called cement to attach permanently to rocks or to other hard substrata. The stalked barnacle Dosima fascicularis is of special interest as it produces a large amount of foam-like cement that can be used as a float. The morphology of the cement apparatus and of the polymerized cement of this species is almost unknown. The current study aims at filling these gaps in our knowledge using light and electron microscopy as well as x-ray microtomography. The shape of the cement gland cells changes from round to ovoid during barnacle development. The cytoplasm of the gland cells, unlike that of some other barnacles, does not have distinct secretory and storage regions. The cement canals, which transport the cement from the gland cells to the base of the stalk, end at different positions in juvenile and mature animals. With increasing size of the cement float, the exit of the cement canals shift from the centrally positioned attachment disk of the vestigial antennules to more lateral positions on the stalk. The bubbles enclosed in the foam-like float are most likely filled with CO(2) that diffuses from the hemolymph into the cement canal system and from there into the cement.

Unusual adhesive production system in the barnacle Lepas anatifera: an ultrastructural and histochemical investigation.

Adhesives that are naturally produced by marine organisms are potential sources of inspiration in the search for medical adhesives. Investigations of barnacle adhesives are at an early stage but it is becoming obvious that barnacles utilize a unique adhesive system compared to other marine organisms. The current study examined the fine structure and chemistry of the glandular system that produces the adhesive of the barnacle Lepas anatifera. All components for the glue originated from large single-cell glands (70-180 µm). Staining (including immunostaining) showed that L-3,4-dihydroxyphenylalanine and phosphoserine were not present in the glue producing tissues, demonstrating that the molecular adhesion of barnacles differs from all other permanently gluing marine animals studied to date. The glandular tissue and adhesive secretion primarily consisted of slightly acidic proteins but also included some carbohydrate. Adhesive proteins were stored in cytoplasmic granules adjacent to an intracellular drainage canal (ICC); observations implicated both merocrine and apocrine mechanisms in the transport of the secretion from the cell cytoplasm to the ICC. Inside the ICC, the secretion was no longer contained within granules but was a flocculent material which became "clumped" as it traveled through the canal network. Hemocytes were not seen within the adhesive "apparatus" (comprising of the glue producing cells and drainage canals), nor was there any structural mechanism by which additions such as hemocytes could be made to the secretion. The unicellular adhesive gland in barnacles is distinct from multicellular adhesive systems observed in marine animals such as mussels and tubeworms. Because the various components are not physically separated in the apparatus, the barnacle adhesive system appears to utilize completely different and unknown mechanisms for maintaining the liquid state of the glue within the body, as well as unidentified mechanisms for the conversion of extruded glue into hard cement.

Old and sticky-adhesive mechanisms in the living fossil Nautilus pompilius (Mollusca, Cephalopoda).

Nautiloidea is the oldest group within the cephalopoda, and modern Nautilus differs much in its outer morphology from all other recent species; its external shell and pinhole camera eye are the most prominent distinguishing characters. A further unique feature of Nautilus within the cephalopods is the lack of suckers or hooks on the tentacles. Instead, the animals use adhesive structures present on the digital tentacles. Earlier studies focused on the general tentacle morphology and put little attention on the adhesive gland system. Our results show that the epithelial parts on the oral adhesive ridge contain three secretory cell types (columnar, goblet, and cell type 1) that differ in shape and granule size. In the non-adhesive aboral epithelium, two glandular cell types (cell types 2 and 3) are present; these were not mentioned in any earlier study and differ from the cells in the adhesive area. The secretory material of all glandular cell types consists mainly of neutral mucopolysaccharide units, whereas one cell type in the non-adhesive epithelium also reacts positive for acidic mucopolysaccharides. The present data indicate that the glue in Nautilus consists mainly of neutral mucopolysaccharides. The glue seems to be a viscous carbohydrate gel, as known from another cephalopod species. De-attachment is apparently effectuated mechanically, i.e., by muscle contraction of the adhesive ridges and tentacle retraction.

Characterization of the adhesive areas in Sepia tuberculata (Mollusca, Cephalopoda).

Adhesion in cephalopods is either mechanical, involving a reduced-pressure system of the arm and tentacle suckers, or is chemically mediated by special adhesive gland structures (as proposed for Euprymna, Idiosepius, and Nautilus). Four species of Sepia (S. typica, S. papillata, S. pulchra, and S. tuberculata) possess grooved structures on the ventral mantle surface and on the fourth arm pair, which are used to attach mechanically to the substratum. Because these areas are often partly covered with sand or debris, it has been hypothesized that chemical substances were involved in this attachment process. This study provides a histochemical and ultrastructural description of the glandular epithelium in the adhesive area of Sepia tuberculata. Two specific glandular cells (Type 1 and Type 2) are present in the epithelium, which differ clearly in their granule size and cellular structure. The aggregation of both cell types and their simultaneous secretion suggest that the secretions of both cell types work synergistically providing a two-component adhesive system which supports the primarily mechanical sucker adhesion by making the arm surface sticky.