Effects of infection with the ectoparasite Argulus japonicus (Thiele) and administration of cortisol on cellular proliferation and apoptosis in the epidermis of common carp,Cyprinus carpio L., skin

The host‐parasite interaction between juvenile carp, Cyprinus carpio, and the ectoparasitic branchiuran, Argulus japonicus, together with the role of cortisol in this interaction, was examined at the level of the host skin epidermis. Epidermal mucous cell numbers, and proliferation and apoptosis of the epithelial cells were studied over 32 days. Apoptotic cell numbers in the uppermost epidermis were reduced at 26 days post‐infection with A. japonicus, while the other parameters were unaffected. Administration of cortisol‐containing food resulted in reduced apoptosis in the cells in the upper skin epidermis at 24 h and at 28 days post‐feeding. Cortisol feeding combined with A. japonicus infection reduced numbers of apoptotic cells in the upper epidermis more than either individual treatment. Further, combining the treatments also significantly increased apoptosis in the lower epidermis in cells morphologically identified as leucocytes apparently migrating macrophages and lymphocytes. Using immunohistochemistry, we demonstrated cortisol receptor presence and cellular localization in the teleost epidermis. Receptors only occurred in pavement cells in the upper epidermis and in leucocytes in the lower parts of the epidermis. The ectoparasites, or administered cortisol, induced effects which may be functionally adaptive in the upper pavement cells, while combining the two treatments also induced changes indicative of immunosuppression.     

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

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.     

On cell kinetics in skin tumours of the Pacific English sole Parophrys vetulus Girard

Abstract. An autoradiographic study of the various types of neoplastic lesions in the skin of Pacific English sole Parophrys vetulus Girard was performed using peritoneally injected tritiated thymidine. The epidermis offish is mitotically active at all cell levels, the most important being the supra-basal layer. In the hyperplastically growing epidermis of the English sole covering connective tissue nodules and in the papillating epidermis prior to formation of enlarged ovoid cells (X-cells), the supra-basal layer also shows the greatest degree of labelling. Parallel to the appearance of these X-cells, which are characteristic of the skin tumours in Pacific flatfish, the distribution of DNA-synthesizing cells is altered. In the tumour areas which are more or less filled with X-cells the 'envelope cells' are labelled throughout with a uniform frequency. On the other hand, the directly adjacent X-cell free areas are labelled with a greater than average frequency.     

褐藻酸降解菌诱发海带病烂的组织细胞学观察

用褐藻酸降解菌对海带进行人为感染，经培养使其产生烂斑，通过组织显微切片观察表明：1、褐藻酸降解菌首先侵入内皮层，然后再逐渐侵入到外皮层、髓部和表皮，最终导致海带病烂。2．感染后内皮层伤害最重，其次是外皮层和髓部，表皮的伤害最轻。3、侵染海皮层的细胞中，有大量颗粒状储藏物质，其直径范围在1．5－4．0μm之间，平均直径为2．7μm。形状为椭圆或圆形。     

短蛸消化系统和部分器官中μ受体的免疫组织化学定位

采用免疫组织化学SABC法对短蛸（Octopus ocellatus）消化系统和部分器官中μ受体（mu opioid receptor）进行定位研究。结果表明,短蛸腕、漏斗、外套膜腹面、口球、嗉囊、胃盲囊、肠、直肠、前唾液腺、后唾液腺、肝胰脏均呈μ受体阴性,但食道内上皮、外膜、结缔组织和胃角质层有μ受体分布,外套膜背面、腕间膜的上皮或近上皮部位呈μ受体阳性。μ受体在短蛸不同部位分布密度的不同,可能与各器官的功能不同有关。     

Pathogenesis and autoradiographic studies of the epidermis of salmonids infested with Ichtyobodo necator (Henneguy, 1883)

Abstract. The sequential pathology of Ichtyobodo necator infestations of the skin of 0+ and 1+ salmon and rainbow trout was studied. Areas of greatest shelter from water currents were found to be most commonly infested and no parasites were found attached to the epidermis on the head of the fish. The parasite caused hyperplasia of the malphigian cells and exhaustion of the goblet cells below infestations, followed by spongiosis of the underlying epidermis. The epidermal plaque then sloughed off leaving a single layer of cells attached to the basement membrane. Cell kinetic studies showed that I. necator caused the cells immediately below infestations to divide, a markedly different pattern from that of normal teleost epidermal cell proliferation. The possibility that the parasite secretes some form of digestive enzyme is postulated. In areas where sloughing had occurred, the remaining malphigian cells were seen to be in the process of division.     

大菱鲆仔鱼皮肤发育及超微结构

The histological development and ultrastructure of larval turbot skin were studied by both light and transmission electron microscopes.The early larvae showed a complete bisymmetry of skin.The skin of 1-25days old larvae developed slowly and was composed of the thin epidermis of 1-2layers epidermal cells and undeveloped dermis before the metamorphosis.But the skin developed significantly when the.metamorphosis began.Till the completion of metamorphosis(about 60days old),the skin contained 3-4layers epidermal cells and very developed collagenous strata.Wisth the process of metamorphosis,the ocular side skin and blind side skin became different in ultrastructure.Ultrastructural observation showed the epidermis of turbot contained three types of epidermal cells;filament-containing cells,mucus cells and chloride cells.The ocular side epidermis with looser structure consisted one type of filament-containing cells,but the blind side epidermis was structured densely and two types of filament-containing cells were observed in it.Melanophores,iridophores,fibroblasts and other cell types and tisssues were found in the spongiosum of dermis.The distribution of melanophores depended on the larval developing stage and the pigmentation of skin.The differentiation of skin seems to be an adaptation for the change of life style of turbot larvae from pelagic living to benthonic living after the metamorphosis.     

短蛸外套膜和足甲硫氨酸脑啡肽的免疫组织化学定位

采用免疫组织化学SABC法对短蛸外套膜和足中甲硫氨酸脑啡肽(met-Enk)进行定位研究。试验结果表明,短蛸外套膜、腕、腕间膜、漏斗中均有met-Enk存在,外套膜背面、腕和腕间膜上皮组织及其附近的部分细胞和神经纤维呈met-Enk强阳性或阳性;外套膜腹面和漏斗上皮组织及其附近的部分细胞和神经纤维呈met-Enk阳性或弱阳性。met-Enk在短蛸外套膜和足不同部位分布密度的不同,可能与各部位的功能不同有关。     

Fluorescence localization of epidermal cathepsins L and B in the Japanese eel

Histochemical localization of proteolytic activities in the dorsal epidermis of Japanese eel was demonstrated by fluorescent microscopy utilizing 4-methoxy-2-naphthylamide (4M$\beta$NA) derivatives as substrates and 5-nitrosalicylaldehyde as a trapping agent. Carbobenzoxy-L-phenylalanyl-L-arginyl-4M$\beta$NA (Cbz-Phe-Arg-4M$\beta$NA) and Cbz-Arg-Arg-4M$\beta$NA were used for direct detection of cathepsins L and B activities, respectively, in fresh frozen sections and unfixed cells of the eel epidermis. The fluorescing areas, where Cbz-Phe-Arg-4M$\beta$NA was hydrolyzed by cathepsin L, were shown in mucus secretory cells and club cells and broadly around skin surface. The fluorescing areas due to Cbz-Arg-Arg-4M$\beta$NA hydrolysis by cathepsin B were localized similarly in these tissues. The fluorescing intensity for both catheptic activities in mucus secretory cells was higher than that in club cells, where small fluorescing granules were distributed. These results indicate that eel cathepsins L and B are stored in epidermal secretory cells at different levels and probably serve as defense factors before or after secretion by these cells. Abbreviations: Cbz – carbobenzoxy; 4M$\beta$NA – 4-methoxy-2-naphthylamide; NSA – 5-nitrosalicylaldehyde.     

六种鲽形目鱼类无眼侧头部皮肤形态特征与功能探讨

本研究采用扫描电镜方法首次对半滑舌鳎、大菱鲆等6种鲽形目鱼类无眼侧头部皮肤形态结构进行观察比较,以期获得与摄食行为等生态习性相关的新发现。结果显示,半滑舌鳎和塞内加尔鳎的无眼侧头部皮肤存在大量游离神经丘,但其形态特征存在差异。半滑舌鳎皮肤表面的游离神经丘类似于乳头状突起,突起上有表皮细胞形成的微脊,多个为一簇,很少单独存在,呈无规则排列;而塞内加尔鳎的游离神经丘直接裸露在皮肤表面,呈线性排列在肉芽状皮肤突起的下方。星突江鲽的头部无眼侧皮肤表面零散分布着多个为一簇的游离神经丘,形状类似于半滑舌鳎的乳头状游离神经丘。大菱鲆、黄盖鲽和条斑星鲽的头部无眼侧皮肤表面未发现游离神经丘等特殊结构,但其表面均凹凸不平。研究推测鲽形目鱼类头部无眼侧皮肤形态特征与其摄食习性密切相关;其头部无眼侧游离神经丘的分布特征与种群分类存在一定的相关性,可为物种的分类提供一定依据。     

Inhibition of epidermal migration in the skin of rainbow trout Salmo gairdneri Richardson in the presence of achromogenic Aeromonas salmonicida

Abstract. Experimental infection of artificially-induced skin ulcers in rainbow trout Salmo gairdneri Richardson with an achromogenic strain of Aeromonas salmonicida induced a syndrome similar to that observed in cyprinid fishes. Sequential histological observations showed that the presence of the bacteria completely inhibited any migration of peripheral epidermal cells and subsequent under-running of the ulcer resulted in its expansion, as peripheral epidermis sloughed.     

Morphopathological features of a severe ulcerative disease outbreak associated with Tenacibaculum maritimum in cultivated sole, Solea senegalensis (L.)

This study describes morphopathologic changes in naturally infected farmed Senegalese sole affected by tenacibaculosis caused by Tenacibaculum maritimum. Macroscopic observation, in addition to light microscopy and scanning electron microscopy, was used to study the lesions. Main lesions were characterized by complete loss of epidermis and dermis, as well as extensive necrosis of muscle layers. Mild-to-moderate inflammatory response with the presence of macrophages was noted around hyaline degenerated muscle cells. Gram-negative filamentous bacteria could be detected only at the dermis. Under scanning electron microscopy, filamentous bacteria located over the scales without epithelium could be observed. These findings together with the isolation and PCR detection of the bacteria in kidney and skin tissues suggest that once the bacteria reach the dermis, probably through eroded epidermis, they are able to proliferate and produce enzymes that are responsible for the damage in the underlying tissues.     

Infection of captive Pagrus auratus (Bloch & Schneider) by the monogenean, Anoplodiscus cirrusspiralis Roubal, Armitage & Rohde (Anoplodiscidae) in Australia

Abstract The monogenean Anoplodiscus cirrtisspiralis infects the fins and nares of snapper, Pagrus auratus (Bloch & Schneider). The epidermis beneath the haptor in both microhabitats is eroded and the parasite attaches to the basement membrane by an adhesive secretion. Captive snapper suffered fin damage through high levels of infection by this parasite. Laboratory experiments showed A. cirrusspiralis to be adversely affected by reduced salinity and killed within 1 h by diluted sea water of < 5%0 salinity.     

The use of chemosterilants to lower the frequency of skin fungal infection amongst precocious male 1+ Atlantic salmon parr, Salmo salar L.

Abstract. Precocious male 1+ Atlantic salmon parr were treated with methallibure and cyproterone acetate during the later stages of spermatogenesis. These drugs prevented hyperplastic changes in the epidermis and lowered the prevalence of fungal infection. They did not alter the mucohistochemical staining pattern or the superficial mucous cell concentration of the skin.     

日本对虾杆状核病毒性传染病诊断技术

本文提出了分别用暗视野显微技术和透射电子显微技术快速确诊日本对虾杆状核病毒性传染病的方法。在暗视野显微镜下观察到的血淋巴中的病毒粒子大小约为0.5μm;胃表层上皮被病毒感染的细胞核明显地呈白色均质小体,直径10～15μm,球形或椭圆形,坏死细胞被囊化为棕色小块(约20～50μm)。病虾血淋巴和胃经负染制样在透射电子显微镜下观察病毒颗粒大小为400×150nm;核衣壳大小为390×85nm。     

Phagocytic cells in the epidermis of rainbow trout, Salmo gairdneri Richardson

Abstract. Electron microscopical observations of skin of rainbow trout revealed the presence of several types of phagocytic cells in the epidermis, most notably macrophages, some of which were seen crossing the basal lamina. Occasionally, filament-containing cells showed either melanin granules or phagosomes, suggesting these cells were also capable of phagocytosis. No cells with the characteristics of the Langerhans cells were observed, which does not necessarily preclude the possibility of macrophages being able to play an identical role as antigen-trapping cells.     

Histopathology caused by Caligus epidemicus Hewitt (Copepoda: Caligidae) on captive Acanthopagrus australis (Günther) (Pisces: Sparidae)

Abstract. Highest densities of copepodids (≤5.3 cm ^-2), chalimus (≤8.5 cm ^-2) and redundant filaments (≤6.7 cm^-2) of Caligus epidemicus were found on the anterodorsal and anteroventral flanks, head and pectoral fins of Acanthopagrus australis (Günther). Copepodids eroded the epidermis and usually attached their frontal filament to the basement membrane on the inter-ray region of fins, as well as the fin rays or scales. Chalimus stages were associated with a healed epidermis and little tissue response. The most extensive tissue response was associated with the redundant frontal filament, and was characterized by infiltrating macrophages, multinucleated giant cells, and epidermal and fibroblast proliferation. No significant pathology could be attributed to either pre-adult or adult copepods.     

Characterization of the pattern of cytokeratin proteins in the epidermal cells of loach,Misgurnus anguillicaudatus (Cyprinformes)

The pattern of cytokeratin proteins in the epidermal cells of loach was studied by immunotechniques and partial separation of the epidermal cells. Two monoclonal antibodies, namely 8F7 and 1C45, against the cytokeratin proteins of the loach epidermis were prepared. these two monoclonal antibodies exhibit distinctive results in immunohistochemical staining. The 8F7 monoclonal antibody stains mainly with the epithelial cells, while the 1C45 monoclonal antibody stains specifically with the club cells. The pattern of cytokeratin proteins in the club cells and the epithelial cells of various epidermal layers was further determined by partial separation of these cells. Immunoblotting analysis of the cell fractions confirms the cytokeratin proteins to be differentially expressed in the club cells and the epithelial cells. However, the cytokeratin proteins expressed in the epithelial cells of the basal, middle and outer layers are same. The results indicate that differentiation of the epithelial cells seems limited during their translocation from basal to upper layers, but in those cells that do differentiate into club cells, the cytokeratin pattern changes.     

The histopathology associated with the juvenile stages of Lepeophtheirus salmonis on the Atlantic salmon, Salmo salar L.

Abstract. Atlantic salmon infected with juvenile stages of Lepeophtheirus salmonis were obtained from sea loch cage sites on the West coast of Scotland. Attached parasites on the skin were studied using light and electron microscopy. Behavioural activity was observed in attached stages maintained on skin in vitro. Pathological changes were associated with four main areas of interaction between the parasite and the host, namely second antennal attachment, filament attachment, maxilliped activity and feeding activity. The skin changes involved were initial mechanical disruption followed by epidermal hyperplasia. The filament material was secreted into the epidermis and spread laterally without breaching the basement membrane. Surprisingly little reaction to the filament occurred, but chalimus activity resulted in chronic changes which were more marked as the later chalimus stages were reached. Fibroplasia was a feature of lesions induced by redundant filaments resulting in small nodular lesions. After erosion of these nodules, all that remained were the melanized rings characteristic of old chalimus sites.     

盐度对瘤背石磺不同部位Na /K -ATP酶活性、围心腔液和腹腔液渗透压及离子含量的影响

为了研究盐度对瘤背石磺不同部位Na+/K+-ATP酶活性、围心腔液和腹腔液渗透压及离子含量(Na+、K+和Cl-)的影响,选取健康的瘤背石磺[湿重(10.24±2.16)g]600只,在不同盐度条件下(5、15、25、35和45)采用模拟生态环境的方式饲养30 d后测定各项指标.结果显示:(1)瘤背石磺腹腔液和围心腔液的盐度和渗透压均随着环境盐度的升高而增高,腹腔液的盐度和渗透压的变动幅度分别为25～ 43和514 ～ 796 mOsm/kg,其增高幅度均大于围心腔液(分别为23 ～ 28和502～ 726 mOsm/kg);(2)围心腔液中Na+、Cl-和K+含量随环境盐度的变化具有一致性,均呈先上升后下降的趋势,且各离子浓度均在盐度25时达到最大值,分别为295.50、322.50和5.15 mmol/L,但盐度对Na+和Cl-含量有显著影响,对K+含量无显著影响.(3)瘤背石磺不同部位的Na +/K+-ATP酶活性具有组织特异性,贲门胃、肠、表皮、肌肉和围心腔膜等部位的Na +/K+-ATP酶活性较高,同时,盐度激活不同部位的Na +/K+-ATP酶活性具有显著差异性,肌肉和贲门胃的Na +/K+-ATP酶在低盐度时活性较高,而肠、肾及围心腔膜等在高盐度时Na+/K+-ATP酶被激活,酶活性较高.研究表明:盐度能显著影响瘤背石磺腹腔液和围心腔液的渗透压和离子浓度以及各器官的Na+/K+-ATP酶活性,贲门胃、肠和肌肉等均是其离子转运和渗透调节的主要部位,具有较强的渗透压调节能力.