Browsing by Author "Ajay Kumar Mittal"

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Alterations in certain immunological parameters in the skin mucus of the carp, Cirrhinus mrigala, infected with the bacteria, Edwardsiella tarda
(Springer Science and Business Media B.V., 2023) Jyoti Singh; Ayan Srivastava; Ashwini Kumar Nigam; Usha Kumari; Swati Mittal; Ajay Kumar Mittal
The bacterial fish pathogen Edwardsiella tarda causes heavy stock mortality, severely hampering fish production, resulting in great economic loss to the farming industry. The first biological barriers that confer immune protection against pathogen entry are the fish mucosal surfaces. The present study was undertaken to investigate the influence of E. tarda on certain enzymatic and non-enzymatic parameters in the skin mucous secretions of the fish Cirrhinus mrigala using spectrophotometry and zymography. Fish were randomly divided into three groups: control, vehicle control, and infected. A sublethal dose of E. tarda (2.2 × 106 CFU/fish) suspended in 50 μL of PBS was injected intra-peritoneally at 0 day (d). Subsequently, mucus samples were collected at 2 d, 4 d, 6 d and 8 d post-infection. The activities of lysozyme (LYZ), protease (PROT), alkaline phosphatase (ALP), acid phosphatase (ACP), catalase (CAT), peroxidase (PER), superoxide dismutase (SOD), and glutathione S-transferase (GST) decreased significantly in the skin mucus of the challenged fish, indicating the suppressed immune system and decreased antioxidant capacity of C. mrigala to E. tarda infection. Lipid peroxidation (LPO) and total nitrate-nitrite were significantly higher at several time points post-infection, suggesting that physiological functions have been impaired following pathogen challenge. The present findings could be relevant for fish aquaculture and underline the importance of skin mucus not only for assessing fish immune status but also for identifying early warning signals of disease caused by pathogens. © 2023, The Author(s), under exclusive licence to Springer Nature B.V.
Alterations in the activity of certain enzymes in the gills of a carp Labeo rohita exposed to an azo dye, Eriochrome black T: a biochemical investigation
(Springer Netherlands, 2018) Ayan Srivastava; Usha Kumari; Ashwini Kumar Nigam; Swati Mittal; Ajay Kumar Mittal
In Labeo rohita exposed to sub-lethal concentrations of an azo dye, Eriochrome black T for 4 days, gills show considerable alterations in the activity of certain metabolic enzymes—alkaline phosphatase, acid phosphatase, carboxylesterase, lactate dehydrogenase, and succinate dehydrogenase; and antioxidant enzymes—catalase and peroxidase. The activities of alkaline phosphatase, acid phosphatase, carboxylesterase, succinate dehydrogenase, catalase, and peroxidase decline significantly. This has been associated with impaired metabolic function of the gills due to azo dye toxicity. The activity of lactate dehydrogenase, in contrast, shows a gradual increase, reflecting a shift from aerobic to anaerobic metabolism. In the fish kept for recovery for 8 days, after exposing the fish to the dye for 4 days, activity of succinate dehydrogenase, alkaline phosphatase, and lactate dehydrogenase gradually become similar to control. Nevertheless, activity of acid phosphatase, catalase, peroxidase, and carboxylesterase, although recover gradually, remained significantly low as compared to that of control. This study signifies that the dye is highly toxic to Labeo rohita and suggests that the activity of metabolic and antioxidant enzymes can be used as biomarker for fish toxicity. © 2017, Springer Science+Business Media B.V., part of Springer Nature.
Alterations in the epidermis and mucus viscosity of the carp, Cirrhinus mrigala, experimentally infected with Edwardsiella tarda
(Academic Press, 2025) Jyoti S. Singh; Usha Kumari; Ravi Prakash; Pralay Maiti; Swati Mittal; Ajay Kumar Mittal
This study investigated effects of a bacterial pathogen, Edwardsiella tarda on the epidermis and mucus viscosity of an Indian major carp, Cirrhinus mrigala. The fish were divided into three groups: a control group (no treatment), a vehicle control group (fish injected with 50 μl of phosphate-buffered saline (PBS) at day 0), and an infected group (fish injected with 50 μl of PBS containing a sublethal dose of 2.2 × 106 CFU/fish, which is 10 % of the 96-h LD50, at day 0). Alterations in the surface architecture, histology, proliferating cell nuclear antigen (PCNA) and inducible nitric oxide synthase (iNOS) expression, specific activity of lactate dehydrogenase (LDH) and succinate dehydrogenase (SDH) were studied at 2d, 4d, 6d and 8d post-infection. Microscopic examination showed hypertrophy of the epidermal epithelial cells, accompanied by disrupted and disorganized microridges, as well as exfoliation. Mucous goblet cells (MGCs) density increased significantly at an early stage of infection. Club cells exhibited degenerative changes, including vacuolization, confluence with neighbouring cells at intervals and simultaneous discharge of their contents onto the surface. A significant increase in iNOS-positive cells was detected. PCNA expression was significantly lower in infected fish, indicating reduced cell proliferation. Cutaneous mucus showed non-Newtonian behavior, with higher viscosity at low shear rates which decreased significantly in infected fish, indicating thinning and shedding under stress. E. tarda infection also caused a significant increase in LDH activity and a decrease in SDH activity. This study will provide deep insight into the host defence mechanisms and serve as a knowledge base for the establishment of early warning systems to control disease outbreaks in farmed fish. © 2025 Elsevier Ltd
Alterations in the epidermis of the carp, Labeo rohita (Cyprinidae: Cypriniformes), infected by the bacteria, Aeromonas hydrophila: A scanning electron microscopic, histopathological and immunohistochemical investigation
(Blackwell Publishing Ltd, 2020) Ayan Srivastava; Arup Mistri; Swati Mittal; Ajay Kumar Mittal
This study was carried out to comprehend the pathogenicity of the bacteria in the epidermis of Labeo rohita inoculated with Aeromonas hydrophila. Alterations in the histopathology of the epidermis were examined using scanning electron microscopy, light microscopy and the localization of iNOS and caspase 3 + ve cells by means of immunohistochemical methods. Skin samples obtained from infected fish at different intervals 2, 4, 6, 8 and 10 days showed significant changes in the cellular components of the epidermis. Epithelial cells often appeared hypertrophied with fragmented and loosely arranged microridges, and in the process of exfoliation. Mucous goblet cells increased significantly in density. Club cells showed degenerative changes, often with simultaneous confluence of adjacent cells and release of their contents. Increase in density of iNOS and caspase 3 + ve cells indicates inflammatory response and apoptosis. This study could provide valuable information on the pathogenesis of the disease, and disease outbreaks in farmed fish. Further, it could provide useful guidelines for fish farmers to take preventive measures for the control of the disease. © 2020 John Wiley & Sons Ltd
Alterations in the skin of Labeo rohita exposed to an azo dye, Eriochrome black T: a histopathological and enzyme biochemical investigation
(Springer Verlag, 2017) Ayan Srivastava; Neeraj Verma; Arup Mistri; Brijesh Ranjan; Ashwini Kumar Nigam; Usha Kumari; Swati Mittal; Ajay Kumar Mittal
Histopathological changes and alterations in the activity of certain metabolic and antioxidant enzymes were analyzed in the head skin of Labeo rohita, exposed to sublethal test concentrations of the azo dye, Eriochrome black T for 4 days, using 24 h renewal bioassay method. Hypertrophied epithelial cells, increased density of mucous goblet cells, and profuse mucous secretion at the surface were considered to protect the skin from toxic impact of the azo dye. Degenerative changes including vacuolization, shrinkage, decrease in dimension, and density of club cells with simultaneous release of their contents in the intercellular spaces were associated to plug them, preventing indiscriminate entry of foreign matter. On exposure of fish to the dye, significant decline in the activity of enzymes—alkaline phosphatase, acid phosphatase, carboxylesterase, succinate dehydrogenase, catalase, and peroxidase—was associated with the binding of dye to the enzymes. Gradual increase in the activity of lactate dehydrogenase was considered to reflect a shift from aerobic to anaerobic metabolism. On transfer of azo dye exposed fish to freshwater, skin gradually recovers and, by 8 days, density and area of mucous goblet cells, club cells, and activity of the enzymes appear similar to that of controls. Alteration in histopathology and enzyme activity could be considered beneficial tool in monitoring environmental toxicity, valuable in the sustenance of fish populations. © 2017, Springer-Verlag Berlin Heidelberg.
Assessment of triclosan induced histopathological and biochemical alterations, and molecular docking simulation analysis of acetylcholinesterase enzyme in the gills of fish, Cyprinus carpio
(Springer, 2024) Anchal Tripathi; Tuhina Gayen; Priyasha Maitra; Usha Kumari; Swati Mittal; Ajay Kumar Mittal
Triclosan (TCS), an antimicrobial additive in various personal and health care products, has been widely detected in aquatic environment around the world. The present study investigated the impacts of TCS in the gills of the fish, Cyprinus carpio employing histopathological, biochemical, molecular docking and simulation analysis. The 96 h LC50 value of TCS in C. carpio was found to be 0.968 mg/L. Fish were exposed to 1/1000th (1 µg/L), 1/100th (10 µg/L), and 1/10th (100 µg/L) of 96 h LC50 value for a period of 28 days. The histopathological alterations observed in the gills were hypertrophy, hyperplasia, edematous swellings, and fusion of secondary lamellae in TCS exposed groups. The severity of these alterations increased with both the concentration as well as the duration of exposure. The present study revealed that the activity of antioxidant enzymes such as superoxide dismutase, catalase, glutathione-S-transferase, glutathione reductase, glutathione peroxidase, and reduced glutathione content decreased significantly (p < 0.05) in both concentration and duration dependent manner. However, a significant (p < 0.05) increase in the activity of the metabolic enzymes such as acid phosphatase and alkaline phosphatase was observed in all three exposure concentrations of TCS from 7 to 28 days. The activity of acetylcholinesterase declined significantly (p < 0.05) from 7 to 28 days whereas the content of acetylcholine increased significantly at the end of 28 day. The experimental results were further confirmed by molecular docking and simulation analysis that showed strong binding of TCS with acetylcholinesterase enzyme. The study revealed that long-term exposure to sublethal concentrations of TCS can lead to severe physiological and histopathological alterations in the fish. © The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2024.
Characterisation of cholinesterases in mucous secretions and their localisation in epidermis of Labeo rohita and Cirrhinus mrigala
(Springer Netherlands, 2019) Ashwini Kumar Nigam; Neeraj Verma; Ayan Srivastava; Usha Kumari; Swati Mittal; Ajay Kumar Mittal
Cholinesterases are multifunctional enzymes and have been associated with diverse physiological functions in addition to their classical role at synapses. In the present study, cholinesterase (ChE) isozymes have been characterised in mucous secretions and their activity has been localised in the epidermis of Labeo rohita and Cirrhinus mrigala. Zymography using specific substrates and inhibitors revealed the presence of two ChE isozymes—ChE-1 and ChE-2. The isozyme ChE-1 was characterised as an atypical butyrylcholinesterase and ChE-2 as a typical acetylcholinesterase in skin mucous secretions of both the fish species. Enzyme histochemical analysis demonstrated the presence of ChE activity in the epidermis of the fish species investigated. In both the fish species, strong ChE activity was observed in the outer-layer epithelial cells, taste buds and neuromasts. The middle and basal layer epithelial cells showed moderate to weak ChE activity. Club cells and mucous goblet cells showed the absence of ChE activity. Characterisation with specific inhibitors indicates that acetylcholinesterase (AChE) was the major cholinesterase type expressed in the epidermis of the two fish species investigated. Immunohistochemical localisation of apoptotic and cell proliferation markers, in addition, revealed high expression of active caspase 3 in the outer-layer epithelial cells, and proliferating cell nuclear antigen (PCNA) in the middle and basal layer epithelial cells. High ChE activity in caspase 3-positive cells in the outer layer of the epidermis and low in PCNA-positive cells in middle and basal layers could point towards the possible involvement of ChEs in cell death and their final extrusion from skin surface. © 2019, Springer Nature B.V.
Characterization of carboxylesterase in skin mucus of Cirrhinus mrigala and its assessment as biomarker of organophosphate exposure
(Kluwer Academic Publishers, 2014) Ashwini Kumar Nigam; Usha Kumari; Swati Mittal; Ajay Kumar Mittal
Presence of carboxylesterase (CbE) activity in the skin mucus of Cirrhinus mrigala was investigated. CbE activity in skin mucus showed higher substrate preference for α-naphthyl acetate over p-nitrophenyl acetate. Four CbE isozymes-CbE-1, CbE-2, CbE-3, and CbE-4 were observed in skin mucus during zymography. The isozyme CbE-4 was characterized as typical serine esterase, whereas CbE-1, CbE-2, and CbE-3 were identified as sulphhydryl group-dependent serine esterases. In vitro treatment of skin mucus with the organophosphorus insecticide, Nuvan® showed strong inhibition of CbE activity. In vivo exposure of the fish to sublethal test concentrations (5 and 15 mg/l) of the insecticide also revealed significant inhibition of CbE activity in mucus. After the cessation of exposure, CbE activity recovered to its control level during the recovery periods. Thus, CbE activity in skin mucus could be considered a biomarker of the organophosphorus insecticide exposure to fish and a useful tool in monitoring environmental toxicity. © 2013 Springer Science+Business Media Dordrecht.
Comparative analysis of innate immune parameters of the skin mucous secretions from certain freshwater teleosts, inhabiting different ecological niches
(Kluwer Academic Publishers, 2012) Ashwini Kumar Nigam; Usha Kumari; Swati Mittal; Ajay Kumar Mittal
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. © 2012 Springer Science+Business Media B.V.
Ecotoxicological impacts of environmentally relevant concentrations of aspirin in the liver of Labeo rohita: Biochemical and histopathological investigation
(Elsevier Ltd, 2023) Tuhina Gayen; Anchal Tripathi; Usha Kumari; Swati Mittal; Ajay Kumar Mittal
Aspirin is one of the emerging pharmaceutical contaminants in the aquatic environment and thus it could impart toxicity to non-target organisms including fish. The present study aims to investigate the biochemical and histopathological alterations in the liver of the fish, Labeo rohita exposed to environmentally relevant concentrations of aspirin (1, 10, and 100 μg/L) for 7, 14, 21, and 28 days. The biochemical investigation revealed a significant (p < 0.05) decrease in the activity of antioxidant enzymes such as catalase, glutathione peroxidase, glutathione reductase; and reduced glutathione content in a concentration and duration dependent manner. Further, the decrease in the activity of superoxide dismutase was in a dose dependent manner. The activity of glutathione-s-transferase, however, increased significantly (p < 0.05) in a dose dependent manner. The lipid peroxidation and total nitrate content showed a significant (p < 0.05) increase in a dose and duration dependent manner. The metabolic enzymes such as acid phosphatase, alkaline phosphatase, and lactate dehydrogenase showed a significant (p < 0.05) increase in all three exposure concentrations and durations. The histopathological alterations in the liver such as vacuolization, hypertrophy of the hepatocytes, nuclear degenerative changes, and bile stagnosis increased in a dose and duration dependent manner. Hence, the present study concludes aspirin has a toxic impact on fish, which is evidenced by its profound effect on biochemical parameters and histopathological analysis. These can be employed as potential indicators of pharmaceutical toxicity in the field of environmental biomonitoring. © 2023 Elsevier Ltd
Effect of asiaticoside on the healing of skin wounds in the carp Cirrhinus mrigala: An immunohistochemical investigation
(Elsevier Ltd, 2017) Neeraj Verma; Usha Kumari; Swati Mittal; Ajay Kumar Mittal
In the present study effect of asiaticoside, on healing of skin wounds in Cirrhinus mrigala is reported. Skin wound, approx. 2 mm in diameter was excised using sterile disposable biopsy punch. Immediately after infliction of the wound, epidermis from wound edge starts migrating as thin sheet toward wound gap. Fronts of migrating epidermis gradually advance, and results in complete epithelialization of wound. Experiments were conducted for 30 days and fishes were divided into control, sham, vehicle control and asiaticoside treated groups. Immunohistochemical localization of proliferating cell nuclear antigen positive cells indicating cellular proliferation and caspase 3 positive cells reflecting apoptosis was carried out and their density at different post wound intervals in each fish group was analyzed. Significant increase in cellular proliferation as well as decrease in apoptosis in both epidermis and dermis in fish treated with asiaticoside compared to sham and vehicle control fish is observed at different intervals of wound repair. This suggests that in treated group healing of skin wounds in fish is enhanced than in sham and vehicle control groups. Asiaticoside treatment in healing of skin wounds would greatly be beneficial to fish farmers as it could protect fish from invasion of pathogens and check fish mortality. © 2017 Elsevier Ltd
Epithelium of lips and associated structures of the Indian major carp, Catla catla
(Springer-Verlag, 1991) Nalini Agrawal; Ajay Kumar Mittal
Structural organization of the epithelium of the lips and associated structures of the Indian major carp, Catla catla, is described. The upper lip is thin and is associated on its dorsal side with a membranous fold of skin and the rostral cap. In contrast, the lower lip is thick and very conspicuous. It is associated on its ventral side with a fold of skin between it and the ventral head skin. The lower lip is divided into a non-projectile portion, a projectile portion and an intermediate groove region. The projectile portion remains folded covering a part of the ventral head skin when the mouth is closed. Their role in relation to the formation of the characteristic feeding tube is discussed. The epithelium of the lips and associated structures is stratified in nature and is composed of the epithelial cells, mucous cells, club cells, lymphocytes and the taste buds. The mucous cells are small, few or even absent and do not appear to secrete profusely at the surfaces of the upper and the lower lips. This suggests that the lips in Catla catla, which feeds on micro-organisms, do not need extra lubrication for protection against abrasion during feeding. In the epithelium at the folds of skin, the voluminous mucous cells secrete profusely and provide extra lubrication to their surface. This reduces the resistance to surface drag during stretching and enables the jaws to protrude with increasing efficiency and swiftness. The club cells are developed additionally to complement the mucous cells in the rostral cap and the upper lip epithelium. Their primary function appears protective in some way, which needs further confirmation. The taste buds, though few in the lower lip, are located in a good number in the upper lip on the characteristic epithelial papillae-like projections, and are projected at the surface. These have been associated with the acute gustatory sense of the fish. The taste buds are absent on the folds of skin where they may not be of much significance. © 1991 The Ichthyological Society of Japan.
Essence of keratin in lips and associated structures of a freshwater fish Puntius sophore in relation to its feeding ecology: Histochemistry and scanning electron microscope investigation
(Elsevier Ltd, 2010) Pinky Tripathi; Ajay Kumar Mittal
Morphological specializations in the lips and associated structures of Puntius sophore were examined by scanning electron microscopy and histochemically. The upper lip (UL), in P. sophore, is associated with the horny upper jaw sheath (HUJS) on its ventral side and with the rostral cap (RC) on its dorsal side through a thin and extensive fold of skin (FSUR). The lower lip (LL) is greatly enlarged, conspicuous and associated with horny lower jaw sheath (HLJS) on the dorsal side and ventrally continues with ventral head skin (VHS). On the lateral sides there is a thin and extensive fold of skin (FSLS) between the lower lip and VHS. In contrast to the mucogenic epithelia of the UL, LL, the RC and fold of skins, the horny jaw sheaths are keratinized in nature and surface epithelial cells are characteristically modified into unculi. The UL and the LL are equipped with epithelial cells (EC), mucous cells (MC) and taste buds (TB) while in addition to these cells club cells (CC) are also present in the RC. Keratin found in unculi is an extremely strong protein which is tough and insoluble, they form the hard but un-mineralized structures. Keratin in unculi could be regarded as an adaptation for browsing or scraping food materials from the substrate as the fish grubs about the bottom. The elaboration of mucus is considered to lubricate the surface and protect the epithelia from abrasions. Taste buds are associated to locate and select palatable food and to trigger a 'pick-up' reflex. © 2010 Elsevier Ltd.
Evaluation of antibacterial activity and innate immune components in skin mucus of Indian major carp, Cirrhinus mrigala
(Blackwell Publishing Ltd, 2017) Ashwini Kumar Nigam; Usha Kumari; Swati Mittal; Ajay Kumar Mittal
The present work has been undertaken to analyse the antibacterial activity and innate immune components in the skin mucus of Indian major carp, Cirrhinus mrigala. Skin mucus was extracted separately in triple-distilled water (TDW), 3% acetic acid (3% AA) or 0.1% trifluoroacetic acid (1% TFA). All mucus extracts exhibited different spectrum of the antibacterial activity against different groups of pathogenic bacteria. Protein profiling by polyacrylamide gel electrophoresis revealed a series of protein bands in the TDW extract, four major protein bands in the AA extract and two protein bands in the TFA extract. Tandem mass spectrometry analysis of distinct protein bands identified potential innate immune factors – histone H2A, histone H3, histone H4, haemoglobin, cofilin and nucleoside diphosphate kinase in the TDW extract, and ubiquitin and histone H2B isoforms in acidic extracts of skin mucus of C.ï¿½mrigala. The presence of these innate immune molecules suggests that skin mucus play an important role in the protection of the fish against microbial invasion. ï¿½ 2015 John Wiley & Sons Ltd
Functional organization of the skin of the 'Green-puffer fish'Tetraodon fluviatilis (Ham.-Buch.) (Tetraodontidae, Pisces)
(Springer-Verlag, 1976) Ajay Kumar Mittal; Tarun Kumar Banerjee
The present study concerns the functional organization of the skin of Tetraodon fluviatilis. The epidermis consists of five different types of cells - the flask-shaped mucous cells, the eosinophilic granular cells, the sacciform granulated cells, the vesicle containing granulated cells, and the polygonal cells. A thin noncellular layer, the cuticle found on the surface of the skin, is probably secreted from the polygonal cells in the outermost layer of the epidermis. A,well-defined lymphatic plexus exists between the cells of the basal layer. Numerous triradiate calcareous spines are embedded within elastic connective tissue pockets in the thick dermis. These pockets are filled with an amorphous, acellular, PAS positive material, and are richly supplied with fine blood capillaries. A histomorphologic basis for the erection of the spines and various structural modifications in the skin facilitating its enormous stretching under inflated conditions of the fish are discussed. © 1976 Springer-Verlag.
Gill epithelium of an angler catfish, Chaca chaca (Siluriformes, Chacidae): Enzyme and glycoprotein histochemistry
(Blackwell Publishing Ltd, 2020) Arup Mistri; Usha Kumari; Swati Mittal; Ajay Kumar Mittal
A series of histochemical techniques have been employed to localize alkaline phosphatase, acid phosphatase, non-specific esterase, catalase and peroxidase; and to visualize and characterize glycoprotein (GPs) moieties in the epithelium of gill arch, gill filaments and secondary lamellae of an angler catfish Chaca chaca. The epithelium of gill arch and gill filament shows strong alkaline phosphatase activity in the deeper layer epithelial cells; strong non-specific esterase activities in the outer layer epithelial cells; and weak acid phosphatase activity throughout the epithelium. The activity of these enzymes in the secondary lamellae is weak. The catalase and peroxidase show strong activities in the blood cells of the secondary lamellae. Various classes of GPs have been identified and characterized in the mucous secretions of the gill epithelium of C. chaca. These include—GPs with oxidizable vicinal diols, GPs with sialic acid residues without O-acyl substitution and GPs with O-sulphate esters. The functional significance of different enzymes in gill epithelium and the GPs in the mucus secreted on the surface has been discussed with the physiology of the gills in relation to the characteristic habit and habitat of the fish. © 2019 Blackwell Verlag GmbH
Healing of cutaneous wounds in a freshwater teleost, Labeo rohita: Scanning electron microscopical investigation
(Wiley-Liss Inc., 2012) Amita Kumari Rai; Nidhi Srivastava; Ashwini Kumar Nigam; Usha Kumari; Swati Mittal; Ajay Kumar Mittal
In this study, healing of cutaneous wounds in Labeo rohita using scanning electron microscope is reported. Wound area could be divided into three regions. Immediately after infliction of wound, edges retract exposing underlying tissues in wound gap (Region I). Simultaneously, at region close to wound edge (Region II), mucous goblet cell openings are observed with copious mucous secretions. Within 1 h, Region I gets covered by mucous secretions, and epidermis at edges starts migrating. Opposing fronts gradually advance and by 4-6 h come in contact to epithelialize wound gap. Zone of contact of fronts is demarcated by epidermal ridge, which is relatively prominent at 8 h. It gradually diminishes and is not distinguished at 24 h and afterward. At 1-4 h, microridges on epithelial cell surfaces appear irregularly arranged, widely spaced, short with abrupt ends at Region I; relatively extensive at Region II; and similar to those in controls at region surrounding Region II (Region III). At 12 h and afterward, microridges appear similar to those in controls at Regions I and II. At 1-2 h, isolated swollen epithelial cells, often in process of detachment and exfoliation at surface, are observed at Regions I and II. Such cells are infrequent at 8 h and afterward. Region I covered by migrated epidermis appears trough like at 4 h to 2 days, level of which gradually rises and at Day 4, surface of epidermis appears at a level similar to that at Regions II and III. Changes have been associated with the imbalance of osmotic homeostasis due to disruption of barrier between internal and external environment of skin. © 2012 Wiley Periodicals, Inc.
HID-AB2.5-PAS and HID-PAS Methods for the Histochemical Analyses of a Variety of Carbohydrates
(1994) Takashi Ueda; Ajay Kumar Mittal; Osamu Fujimori
The high iron diamine (HID) staining procedure was combined with alcian blue pH 2.5 (AB2.5) and periodic acid-Schiff (PAS) methods for the simultaneous demonstration of carbohydrates containing sulfate esters, carboxyl groups and oxidizable vicinal diols, whereas it was sequentially employed with PAS alone to differentiate sulfate esters from oxidizable vicinal diols. A variety of rat tissues and the epidermis of four fish species were utilized to test the specificity or selectivity of these methods. The HID-AB2.5-PAS sequence largely coloured carbohydrates containing sulfate esters, carboxyl groups and oxidizable vicinal diols in brownish black, turquoise and magenta shades respectively. The HID-PAS sequence coloured sulfate esters brownish black and oxidizable vicinal diols magenta. On the basis of the experimental and control studies on the both staining techniques, these staining methods could be postulated to represent efficient and useful techniques for precise histochemical analyses and simultaneous differentiations of a variety of carbohydrates in light microscopy. © 1994, Editorial Board of Okajimas Folia Anatomica Japonica. All rights reserved.
Histochemical analysis of glycoproteins in the gill epithelium of an Indian major carp, Cirrhinus mrigala
(2012) Nidhi Srivastava; Usha Kumari; Amita Kumari Rai; Swati Mittal; Ajay Kumar Mittal
Glycoproteins were analyzed by a range of histochemical methods in the epithelium of gills of . Cirrhinus mrigala, a valuable food fish of great economic importance cultured extensively in India. The gills consist of gill arches, gill rakers, gill filaments and secondary lamellae. Major components of the epithelium of gill arches and gill rakers are epithelial cells, mucous goblet cells, rodlet cells, lymphocytes, eosinophilic granular cells and taste buds. In contrast, in the gill filament epithelium, rodlet cells and taste buds, and in secondary lamellae epithelium, rodlet cells, lymphocytes, eosinophilic granular cells and taste buds are not discernible. The epithelial cells, the mucous goblet cells and the eosinophilic granular cells elaborate glycoproteins with oxidizable vicinal diols and glycoproteins with sialic acid residues without O-acyl substitution. In addition, glycoproteins with O-sulphate esters are secreted by the mucous goblet cells. The rodlet cells elaborate glycoproteins with oxidizable vicinal diols. Different types of glycoproteins elaborated on the epithelial surface of gills are discussed in relation to physiological significance of glycoprotein classes with special reference to their roles in lubrication, protection and inhibition of invasion and proliferation of pathogenic micro-organisms. © 2011 Elsevier GmbH.
Histochemical analysis of glycoproteins in the secretory cells in the gill epithelium of a catfish, Rita rita (Siluriformes, Bagridae)
(Elsevier Ltd, 2009) Usha Kumari; Madhu Yashpal; Swati Mittal; Ajay Kumar Mittal
Glycoproteins (GPs) were visualised histochemically in the secretory cells - the mucous goblet cells (the type A and the type B), the serous goblet cells, the club cells and the epithelial cells in the gill epithelium of Rita rita. The type A mucous goblet cells, the type B mucous goblet cells and the epithelial cells elaborate GPs with oxidizable vicinal diols and GPs with sialic acid residue without O-acyl substitution. In addition, GPs with O-sulphate esters are elaborated by the type A and GPs with O-acyl sugars by the type B mucous goblet cells. GPs are absent in the serous goblet cells and are with oxidizable vicinal diols in low moieties in the club cells. The analysis of the results elucidates interesting differences in the composition and concentration of GPs in the mucus elaborated by the epithelium of the gill arches and the gill rakers; and the gill filaments and the secondary lamellae indicating the potential importance of the glycoproteins at these locations. GPs elaborated on the surfaces of the gill arches and the gill rakers could be associated to assist in feeding activities and on the surfaces of the gill filaments and the secondary lamellae in the respiratory activity. © 2009 Elsevier Ltd. All rights reserved.