The innate immune system of fish is considered first line of defense against a broad spectrum of pathogens. Being a component of innate immunity and lying at the interface between fish and the aqueous environment, skin mucus plays a frontier role in protecting fish from infections. In the present study, skin mucus of Cirrhinus mrigala, Labeo rohita, Catla catla, Rita rita and Channa punctata, inhabiting different ecological niches, was analyzed to characterize potential innate immune factors such as lysozyme, proteases, phosphatases, esterase and sialic acid. The enzyme activities were high in bottom dweller species, C. punctata and C. mrigala, and low in clean water inhabiting species, L. rohita and C. catla. An inverse relationship was observed between the level of enzyme activity and the sialic acid content in these fish species. In R. rita, however, the levels of all factors were found to be low. Zymographic analysis with labeled Micrococcus lysodeikticus revealed three isoforms of lysozyme in C. punctata and two in each species, C. mrigala, L. rohita and C. catla. In R. rita, lysozyme could not be detected. Gelatin zymography revealed that serine and metalloproteases were the major mucus proteases in all fish species investigated. In addition, trypsin-like protease and Ca(++)-specific serine proteases were observed in skin mucus. Increased knowledge of these parameters could be useful in understanding the role of skin mucus in the innate immune system of fish species inhabiting different ecological niches.
The buccal cavity of an herbivorous fish, Cirrhinus mrigala, was investigated by scanning electron microscopy to determine its surface ultrastructure. The buccal cavity shows significant adaptive modifications in relation to food and feeding ecology of the fish. The buccal cavity of the fish is of modest size and limited capacity, which is considered an adaptation with respect to the small-sized food items primarily consumed by the fish that could be accommodated in a small space. Modification of surface epithelial cells, on the upper jaw, into characteristic structures-the unculi-is considered an adaptation to browse or scrap, to grasp food materials, e.g., algal felts, and to protect the epithelial surface against abrasions, likely to occur during their characteristic feeding behavior. Differentiation of the highly specialized lamellar organ on the anterior region of the palate could be an adaptation playing a significant role in the selection, retention, and sorting out of palatable food particles from the unpalatable items ingested by the fish. The filamentous epithelial projections and the lingulate epithelial projections on the palatal organ in the posterior region of the palate are considered to serve a critical function in final selection, handling, maneuvering, and propelling the food particles toward the esophagus. The abundance of different categories of taste buds in the buccal cavity suggests that gustation is well developed and the fish is highly responsive in the evaluation and the selection of the preferred palatable food items. The secretions of mucous cells in the buccal cavity are associated with multiple functions-particle entrapment, lubrication of the buccal epithelium and food particles to assist smooth passage of food, and to protect the epithelium from possible abrasion. These morphological characteristics ensure efficient working of the buccal cavity in the assessment of the quality and palatability of ingested food, their retention and transport toward the esophagus. Such an adaptation may be essential in conducting the function most basic to the survival of the individuals and species-feeding.
The unicellular glands in the epidermis of the Indian freshwater fish Mastacembelus pancalus consist of three types of mucous cells and sacciform cells. The histochemical properties of their secretory glycoproteins have been analysed by means of a battery of histochemical methods. These included methods for the identification and simultaneous visualization of oxidizable vicinal diols, O-acyl sugars, O-sulphate esters and sialic acid residues with or without side-chain O-acyl variants. Four general classes of glycoproteins (GPs) were identified. These included (i) GPs with O-sulphate esters and oxidizable vicinal diols, (ii) GPs with oxidizable vicinal diols and sialic acid residues with or without O-acyl substitution at C7, (iii) GPs mainly with O-sulphate esters, low moieties of GPs with oxidizable vicinal diols, O-acyl sugars and sialic acid residues with side-chain O-acyl variant predominantly at C8 (or which are di- or tri-substituted) or C9 and in traces of sialic acid residues without O-acyl substitution or with O-acyl substitution at C7, and (iv) GPs with traces of oxidizable vicinal diols, O-acyl sugars and sialic acid residues with O-acyl substitution at C8 (or which are di- or tri-substituted) or C9. The physiological significances of these GP classes and their release on the surface of the epidermis are discussed with special reference to their role in lubrication, protection and inhibition of the invasion and proliferation of pathogenic microorganisms in the epidermis, as adapted to the peculiar mode of life of the fish.
Gill arches and the gill rakers of a sluggish, carnivorous catfish, Rita rita, show significant differences of their surface ultrastructure, which are recognized adaptive modifications in relation to food and feeding ecology of fish. Gill rakers on the first and second pairs of gill arches are borne on the oral side and are long and stout at the epi-ceratobranchial union. Gill rakers on the third and fourth pairs of gill arches, in contrast, are borne on the oral and aboral sides and are relatively delicate and short. Long and stout gill rakers on the first and second pairs of gill arches are considered primarily to prevent entry of undesirably large food items into the pharynx. Two types of taste buds, Type I and Type II, occur on the gill arches and the gill rakers. The raised taste buds, located at the apical ends of the gill rakers on the third, fourth, and the fifth pairs of gill arches could increase gustatory efficiency in the pharynx. Differences in the distribution of taste buds on the pharyngeal sides of different gill arches indicate that the posterior part of the pharynx plays a more crucial role in gustation than does the anterior part. Co-occurrence of teeth and taste buds on the epi- and hypopharyngeal bones denotes that food processing and gustation occur simultaneously in the pharynx. Villiform and caniform teeth on the epi- and hypopharyngeal bones are associated with a complex food-processing cycle. Mucous secretions, oozing through mucous cell openings, provide lubrication facilitating smooth passage of food through the pharynx. The angle of curvature at the epi-ceratobranchial union of the first to fourth pairs of gill arches could assist the ventral drag of ceratobranchials in lowering of the pharyngeal floor, thus resulting in a great expansion of the pharynx, as needed to accommodate the large quantities of food captured.
Mucus is secreted to the surface of the body and fin webs of Blennius pholis by superficial epithelial cells and by goblet cells. Some goblet cells secrete sulphated acid glycoproteins, others produce a mucus which is neutral or mixed in its reactions. The superficial epithelial cells of these areas secrete sulphated acid glycoproteins, seen by electron microscopy as electron-lucent or moderately lucent vesicles; this secretion is not normally visible external to the skin in transmission electron microscope (TEM) sections. These cells do not react to the bromphenol blue test for proteins. Over part of the surface of the pelvic fins and the distal parts of the rays of the pectoral fins, the skin contains no goblet cells and bears a thick external secretion, or cuticle, containing protein and glycoprotein which is mainly neutral in reaction, although some cells at the edges of the region secrete weakly sulphated or non-sulphated acidic glycoprotein. The protein content of the columnar superficial epithelial cells of these regions correlates with the fibrous nature of the secreted cuticular layer as seen by TEM; the columnar cells are characterized by extensive ribosomal endoplasmic reticulum and vesicles which stain darkly with phosphotungstic acid, less so with uranyl acetate. The distal part of the cell, containing these vesicles, reacts positively to the PAS stain. In some places the borders of the zones with fibrous cuticle are characterized by cuboidal superficial epithelial cells which give a strong positive reaction to alcian blue at pH I .O, indicating the presence of sulphated acid glycoproteins, but also react positively to the bromphenol blue test for proteins.
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