Ultrastructural characterization of gills in juveniles of the Argentinian Silverside, Odontesthes bonariensis (Valenciennes, 1835) (Teleostei: Atheriniformes)

An ultrastructural study was performed on the gills of juvenile Argentinian silverside, Odontesthes bonariensis. The gills are composed of two sets of four holobranchs and, in turn, each holobranch consists of a gill arch and two rows of caudolaterally projecting branchial filaments. From the dorsal and ventral surfaces of each filament, branchial lamellae radiate out as foldings of the epithelial layer. Gill rakers are present on each of the gill arches, on the anteromedial side of the arch opposite to the filaments. Gill rakers, gill arches and branchial filaments are covered by a stratified epithelium, whereas branchial lamellae essentially consist of a thin epithelial envelope containing capillaries. In the stratified epithelium, mucous cells, rodlet cells, granular cells, pavement epithelial cells and mitochondria-rich cells are identified. The thin epithelium that lines the lamellae comprises two cell types, outer and inner epithelial cells, and the capillary walls on the inside of the epithelial envelope are defined by pillar cells. The ultrastructure of all these cell types is described and our findings are discussed in light of the existing data on fish gill morphology. In the gills of juvenile Argentinian silverside is of particular interest the characteristics showed by mitochondria-rich cells, such as their arrangement in clusters of 2-3 cells and their small and depressed surface in contact with the aquatic milieu, features which strongly resemble those of euryhaline species.     

The Effect of Copper from Industrial Emission on the Seminiferous Epithelium in Rams

Contents: The changes in the testes of adult rams have been followed by methods of light-and electronmicroscopy after intoxication with copper emitted from a copper plant. In the toxic phase of the disease primarily the germinative epithelium was damaged. The breaking or decay of the cellular membrane with consequent devastation or stagnation of the spermiogenesis was observed. The toxic effect of the copper affects irreversibly not only all the cells of spermiogenesis in the developmental stages, but it damages also the Sertoli cells including the components of the interstitium. These changes participated in the impairment of reproduction in the flock under study.     

An outbreak of nephropathogenic H13 infectious bronchitis in commercial broilers

A nephropathogenic Massachusetts strain of infectious bronchitis virus, designated H13-IB virus, was isolated from the kidneys of broilers in a commercial flock. Respiratory distress, diarrhea, depression, and high mortality were present. Gross renal lesions consisted of pale coloration, mottling, urate deposition, and swelling. The trachea contained a serous to catarrhal exudate. Microscopically, renal changes were primarily associated with the medullary region. Lesions included an interstitial mononuclear cell infiltration, urate deposition, edema, and degenerative and necrotic tubular changes. Tracheal changes included marked thickening of the mucosa, desquamation of ciliated and glandular epithelia and replacement with a stratified undifferentiated epithelium, and inflammatory cell infiltration.     

剑尾鱼硫酸铜急性中毒的病理学研究

通过对实验动物化的剑尾鱼进行急性硫酸铜中毒实验,在观察症状和剖检变化以后,对实验96h期间死亡鱼与96h后仍存活的鱼进行石蜡切片,HE染色,光学显微镜观察。主要眼观病变为肝脏水肿和白色坏死点;鳃丝肿胀,黏液分泌增多;肾脏肿胀,充血。显微病变为肝细胞水泡变性,严重坏死;鳃小片上皮细胞肿胀,脱落严重;有的鳃小片发生融合,整个鳃丝呈棒状化;肾小管上皮细胞空泡变性,间质有充血或网织细胞增多,细胞间噬铁血红素细胞增多;肝胰腺的胰腺细胞的酶原颗粒减少。     

Structural and Ultrastructural Differentiation of Cell Types in the Gills of the Tench (Tinca tinca, L.)

This study used 22 tench of 25-30 cm in length and weighing roughly 200 g. After sacrifice, the gill arch was extracted and fixed in 5% glutaraldehyde for structural, ultrastructural and morphometrical study. Tench gills consist of primary lamellae which are perpendicular to numerous secondary lamellae in which the respiratory barrier is located. A study was made of the structures which make up that barrier and of the multi-layered epithelium situated in the interlamellar space, which is closely linked to maintaining of internal balance in the fish.     

Clinical and pathological studies on intoxication in horses from freshly cut Jimson weed (Datura stramonium)-contaminated maize intended for ensiling

Spontaneous intoxication in 34 horses after ingesting freshly harvested maize that was to be used for ensiling and heavily contaminated with young Datura stramonium plants, is described. The clinical status of all horses was monitored for 7 days, and included body (rectal) temperature, respiratory and heart rates, colour and moistness of visible mucosae, changes in pupil size, appetite, thirst, general behaviour, locomotion, sensory perceptions, urination and defaecation. The intoxication was accompanied by altered clinical status, namely mild hyperthermia, tachycardia, polypnoea, dyspnoea and shallow breathing, mydriasis, dry oral, rectal, vaginal and nasal mucosae, acute gastric dilatation and severe intestinal gas accumulation, anorexia to complete refusal of feed, decreased or absent thirst, absence of defaecation and urination. As a result of the treatment, the clinical parameters normalised between days 2 and 5. Necropsies and pathological studies were performed on two horses that died, revealing toxic liver dystrophy, cardiac lesions and substantial dystrophic and necrotic processes in the kidneys. The observed clinical signs, the pathomorphological changes and the applied therapy could be used in the diagnosis, differential diagnosis, prognosis and treatment of Jimson weed intoxication.     

Gill Infection Model for Columnaris Disease in Common Carp and Rainbow Trout

Challenge models generating gill lesions typical for columnaris disease were developed for the fry of both Common Carp Cyprinus carpio and Rainbow Trout Oncorhynchus mykiss by means of an immersion challenge and Flavobacterium columnare field isolates were characterized regarding virulence. Carp inoculated with highly virulent isolates revealed diffuse, whitish discoloration of the gills affecting all arches, while in trout mostly unilateral focal lesions, which were restricted to the first two gill arches, occurred. Light microscopic examination of the gills of carp exposed to highly virulent isolates revealed a diffuse loss of branchial structures and desquamation and necrosis of gill epithelium with fusion of filaments and lamellae. In severe cases, large parts of the filaments were replaced with necrotic debris entangled with massive clusters of F. columnare bacterial cells enwrapped in an eosinophilic matrix. In trout, histopathologic lesions were similar but less extensive and much more focal, and well delineated from apparently healthy tissue. Scanning and transmission electron microscopic observations of the affected gills showed long, slender bacterial cells contained in an extracellular matrix and in close contact with the destructed gill tissue.

This is the first study to reveal gill lesions typical for columnaris disease at macroscopic, light microscopic, and ultrastructural levels in both Common Carp and Rainbow Trout following a challenge with F. columnare. The results provide a basis for research opportunities to examine pathogen–gill interactions.

Received January 10, 2014; accepted July 27, 2014     

MHC class II+ cells in the gills of Atlantic salmon (Salmo salar L.) affected by amoebic gill disease

Amoebic gill disease (AGD) is characterised by the association of Neoparamoeba sp. with hyperplastic gill tissue of affected fishes, however, the identity and role of host cells associated with AGD lesions are not known. Here, we investigated cells with an immunological role that were associated with AGD lesions by locating cellular MHC class II β chain. A tank housing Atlantic salmon (Salmo salar) was inoculated with Neoparamoeba sp., and MHC class II β chain expression in the gills was qualitatively assessed by immunohistochemistry. In AGD-naïve control fish, MHC class II⁺ cells were detected basolateral to the interlamellar epithelium as well as upon the interlamellar and secondary epithelium. In the gills of AGD affected fish MHC class II⁺ cells were observed in both affected and unaffected tissue. Within AGD lesions, numerous MHC class II⁺ cells were present and these cells exhibited variable levels of expression suggesting that like mammals, MHC class II expression is highly regulated. The presence of MHC class II⁺ cells within gill lesions is indicative of immune cell trafficking and these cells could contribute in an antigen presentation capacity to the development of an antibody response in fish chronically affected by AGD.     

Gill epithelium of an angler catfish,Chaca chaca (Siluriformes,Chacidae): Enzyme and glycoprotein histochemistry

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.     

Sub-clinical necrotic enteritis in broiler chickens: Novel etiological consideration based on ultra-structural and molecular changes in the intestinal tissue

The present study revealed several previously not recognized etiological details in the development of necrotic enteritis (NE) in broilers. We provide evidence that the pathological process leading to mucosal epithelium necrosis follows morphologically distinct phases commencing at the basal domain of the mucosal epithelium and then progressively invading the entire lamina propria. Initially mucosal epithelium appears normal, but as the pathological changes progress throughout the lamina propria, the adjacent enterocytes begin to show features of necrotic cell death and the necrotic process of the epithelium progresses from being focal to locally extensive.Ultra-structural examination showed that primary changes occur at the level of basal and lateral domains of the enterocytes, whereas the apical domain of enterocytes remains intact even in the face of advanced necrotic changes. This indicates that the mucosal necrosis does not result from direct damage to the mucosal epithelium. Rather, the necrotic death of enterocytes is a consequential effect of the destruction of lamina propria, the extra-cellular matrix, and intercellular junctions.The nature of these morphological changes indicates that initiation of the pathological process leading to NE involves proteolytic factors affecting the extra-cellular matrix and cellular junctions. Further studies revealed that, indeed, the elevated activity of collagenolytic enzymes in the mucosal milieu and in intestinal tissue represents an integral component of the pathological process leading to NE. In the first instance we discovered that Clostridium perfringens strains isolated from field cases of NE secrete several potent collagenolytic enzymes. In the second instance we observed that, in comparison to controls, broilers challenged with C. perfringens isolated from field cases of NE show high levels of several collagenolytic enzymes in the intestinal tissue. A major component of the overall collagenolytic activity detected in the intestinal tissue was identified by zymography as matrix metalloproteinases (MMPs). Dominant activity was associated with MMP-2. We confirmed using immuno-histochemistry that this enzyme is expressed at high levels in mucosal tissue showing signs of NE.The high levels of collagenolytic activities, in particular associated with MMP-2, demonstrated in our studies are consistent with the nature of morphological changes observed primarily in extra-cellular matrix (ECM) at the basal domain of enterocytes, as well lateral domains of enterocytes. The lack of changes at the level of apical domain of mucosal epithelium indicates that the lipolytic aspect of alpha toxin in NE is not an essential factor in primary lesions development. Taken together, our findings indicate that the early lesions leading to NE are associated with virulence factors that induce proteolytic activity, rather than lipolytic activity.     

Sub-clinical necrotic enteritis in broiler chickens: novel etiological consideration based on ultra-structural and molecular changes in the intestinal tissue

The present study revealed several previously not recognized etiological details in the development of necrotic enteritis (NE) in broilers. We provide evidence that the pathological process leading to mucosal epithelium necrosis follows morphologically distinct phases commencing at the basal domain of the mucosal epithelium and then progressively invading the entire lamina propria. Initially mucosal epithelium appears normal, but as the pathological changes progress throughout the lamina propria, the adjacent enterocytes begin to show features of necrotic cell death and the necrotic process of the epithelium progresses from being focal to locally extensive.Ultra-structural examination showed that primary changes occur at the level of basal and lateral domains of the enterocytes, whereas the apical domain of enterocytes remains intact even in the face of advanced necrotic changes. This indicates that the mucosal necrosis does not result from direct damage to the mucosal epithelium. Rather, the necrotic death of enterocytes is a consequential effect of the destruction of lamina propria, the extra-cellular matrix, and intercellular junctions.The nature of these morphological changes indicates that initiation of the pathological process leading to NE involves proteolytic factors affecting the extra-cellular matrix and cellular junctions. Further studies revealed that, indeed, the elevated activity of collagenolytic enzymes in the mucosal milieu and in intestinal tissue represents an integral component of the pathological process leading to NE. In the first instance we discovered that Clostridium perfringens strains isolated from field cases of NE secrete several potent collagenolytic enzymes. In the second instance we observed that, in comparison to controls, broilers challenged with C. perfringens isolated from field cases of NE show high levels of several collagenolytic enzymes in the intestinal tissue. A major component of the overall collagenolytic activity detected in the intestinal tissue was identified by zymography as matrix metalloproteinases (MMPs). Dominant activity was associated with MMP-2. We confirmed using immuno-histochemistry that this enzyme is expressed at high levels in mucosal tissue showing signs of NE.The high levels of collagenolytic activities, in particular associated with MMP-2, demonstrated in our studies are consistent with the nature of morphological changes observed primarily in extra-cellular matrix (ECM) at the basal domain of enterocytes, as well lateral domains of enterocytes. The lack of changes at the level of apical domain of mucosal epithelium indicates that the lipolytic aspect of alpha toxin in NE is not an essential factor in primary lesions development. Taken together, our findings indicate that the early lesions leading to NE are associated with virulence factors that induce proteolytic activity, rather than lipolytic activity.     

Clinical and pathological features of common gill diseases of cultured salmonids in Ontario

We reviewed the clinical presentations and histopathology of 118 diagnostic submissions of trout with infectious gill diseases from commercial trout farms within Ontario. Bacterial gill disease (BGD) (56%) and nodular gill disease (NGD) (26.2%) together accounted for 82.2% of these submissions. Submissions of fish with BGD occurred in every month of the year, but were proportionally more frequent in late winter and spring than in late summer and fall. Chemotherapy of groups of fish with gill diseases was common prior to submission, but the success rate of such treatment was low (29% for BGD; 21% for NGD). Specific therapeutic protocols implemented following etiological diagnosis of BGD were successful in 80% of the previously unresponsive cases and in 88.8% of previously untreated cases. The gills of trout collected within 48-96 h of treatment and apparent clinical recovery lacked bacteria and necrotic epithelial cells, but features such as lamellar fusion and epithelial hyperplasia were similar between recovered and affected fish.     

PRRSV感染仔猪肾组织的超微病理变化分析

猪繁殖与呼吸综合征病毒（PRRSV）是能引起仔猪高死亡率的疾病,给养猪业带来极大的经济损失。本研究对PRRSV感染仔猪肾组织的病毒定位及产生的超微病理变化进行观察,以期了解PRRSV对肾组织细胞以及血液循环的影响。主要采用透射电镜技术结合血清生化检测、免疫组织化学染色（IHC）,对仔猪感染PRRSV后肾组织进行超微病理学观察及分析。结果发现,PRRSV感染后引起肾组织损伤,病毒主要分布于肾小球、坏死的肾小管上皮细胞胞质及巨噬细胞胞质内;肾小球内足细胞足突广泛融合或微绒毛化,内皮细胞肿胀;肾小管上皮细胞线粒体肿胀、嵴断裂、溶解;间质炎性细胞浸润。PRRSV通过巨噬细胞内复制随血液扩散至肾组织,引起损伤;根据透射电镜观察结果,足细胞足突融合,肾小球血管内皮细胞损伤引起微血栓形成,说明PRRSV感染仔猪后影响了肾小球滤过率,同时,对肾血液微循环系统造成损伤。本研究为阐明PRRSV的感染致病机制提供理论基础。     

Occurrence and pathology of Sinergasilus lieni (Copepoda: Ergasilidae), a parasite of the silver carp and bighead, in Hungarian ponds

Sinergasilus lieni Yin, 1949, a well-known and pathogenic parasitic copepod in China and Russia, has been detected in Hungarian carp farms for the first time. The parasite infected the third-year generation of silver carp (Hypophthalmichthys molitrix) and bighead (Aristichthys nobilis). The gills of the infected fish specimens showed severe pathological changes. At the attachment sites of female copepods clubbing and fusing of the gill filaments were observed and in some parts of the pale or whitish hemibranchia deep indentations were recorded in places where the tips of the damaged filaments had broken off. Silver carp and bighead were infected at a similar rate, having 8 to 27 copepods attached to the end of the clubbed filaments or the proliferated epithelium of 2 to 10 fused filaments. In histological sections the head part of the parasite was found in a deep cavity of the proliferated epithelium, piercing its antennae deep into the tissues. Only the end of the filaments showed changes. In this part the proliferated epithelium was infiltrated by eosinophilic granular cells. In the central and basal parts of the hemibranchia the original structure of the filaments was preserved with intact secondary lamellae.     

Myxobolus infection of the gills of common bream (Abramis brama L.) in Lake Balaton and in the Kis-Balaton reservoir, Hungary

During a five-year survey including studies on the parasite fauna of bream (Abramis brama), four gill-parasitic Myxobolus species (M. bramae, M. hungaricus, M. impressus and M. macrocapsularis) were recorded in a total of 313 breams from Lake Balaton. The commonest species, M. bramae showed a prevalence of 33%, while the other species occurred sporadically. Myxobolus bramae and M. macrocapsularis infected the tips of the gill filaments and caused both intralamellar and interlamellar infection. Intralamellar plasmodia of small size developed in the capillary network of the gill lamellae whereas the much larger interlamellar plasmodia were formed in the arteria afferens. The intralamellar plasmodia of M. hungaricus always infected the basal or central part of the gill filaments. In contrast to the above species developing in the blood vessels, M. impressus proved to be an epithelial parasite, as its plasmodia always developed in the adjacent gill filaments of two opposite haemibranchia, in the stratified epithelium between the respiratory plates, causing changes of the haemibranchium which were well visible even by the naked eye.     

Induction and Evaluation of Proliferative Gill Disease in Channel Catfish Fingerlings

Abstract

Proliferative gill disease (PGD) was first reported in channel catfish Ictalurus punctatus at commercial farms in 1981 and is caused by the myxozoan parasite Henneguya ictaluri. The disease affects the gills and is characterized by severe branchial inflammation, epithelial hyperplasia, lamellar fusion, and lysis of chondrocytes. Presumptive diagnosis is based on the presence of lytic areas in the cartilage of the primary lamellae on microscopic examination and is confirmed histologically by the presence of the organism. In these trials, PGD was induced by exposing channel catfish fingerlings to fresh or aged infectious water collected from a pond containing fish diagnosed with severe PGD. The severity of disease was graded by histological scoring and microscopic examination of wet mounts to determine the percentage of gill filaments containing chondrolytic lesions. Exposure of fish to infectious pond water was shown to produce pathological lesions consistent with PGD, and the percentage of gill filaments containing chondrolytic lesions was positively correlated with histological scoring of gill pathology. The number of trophozoite stages in the gills was shown to increase with the severity of the disease. In most cases, however, parasitic cells were not observed in tissue samples with chondrolytic lesions during the early stages of infection. These observations indicate that pathology and lysis of chondrocytes can occur prior to detection of the organism by histopathology. Exposing fish to infectious pond water that was aged for 1 d produced negligible gill pathology and implies that the infectivity of the H. ictaluri actinospore stage is short lived. Removing fish from the source of infection promoted repair of damaged gill tissue; within 14 d of fish transfer to clean water, gill pathology associated with the acute infection was negligible.     

Aspects of the pathology of Lamproglena clariae (Copepoda: Lernaeidae) on gills of Clarias gariepinus from the Vaal River system,South Africa

Lamproglena clariae attaches to gill filaments of Clarias gariepinus, penetrates the gill tissue and consumes blood. Hence, the purpose of this study was to investigate the mechanical damage caused by this parasite on gill filaments of the host. Infestation intensity and haematocrit values of fish blood were determined and correlated. Fish were killed, gills dissected out and observed with a dissection microscope. Gill filaments with adult females in situ were fixed in a solution of alcohol, formaldehyde and acetic acid and preserved in 70% ethanol. Fixed specimens were studied by histological examination following staining with Heidenhein’s azan solution. Gross morphology revealed varying degrees of swelling of gill filaments near the attached specimens of L. clariae. Epithelial hyperplasia, resulting in fusion of gill lamellae and necrosis of the host tissue in the vicinity of the parasite were revealed through histological examination. No correlation was evident between the infestation intensity and fish haematocrit values. Although results showed that L. clariae causes localized infestation, high intensity of infestation may be detrimental to the fish. Hyperplasia of epithelial tissue resulting in fusion of lamellae would result in restriction of air passages and thus hinder the process of respiration of the host.     

Pathological changes in the mammary gland and biochemical changes in milk of the goat following oral dosing with leaf of the Avocado (Persea americana)

Two varieties of avocado leaves (Persea americana var Guatemalan and var Mexican) were administered to lactating goats. The Mexican variety was without effect. The Guatemalan variety in doses exceeding 20 g fresh leaf per kg bodyweight, produced damage to the mammary gland with decreased milk production. The lesions were characterised by oedema and reddening, with clots in the large ducts. Microscopically, there was widespread degeneration and necrosis of the secretory epithelium, the necrotic cells sloughing into the lumen. There was no significant cellular inflammatory response. Concentrations of antitrypsin in the milk, indicating changes in vascular permeability, increased rapidly 15 h after a single high dose, coinciding with palpable oedema. Concentrations of NAGase, indicating cell damage, increased after 24 h. Goats given multiple doses followed a similar pattern but the initial response was delayed. The toxic principle, and its mode of action in selectively damaging mammary secretory cells, remains to be determined.     

Morphological identification of cryptosporidia in the intestines of calves using light and scanning electron microscopy

Small and large intestines of ten calves of the age from 7 to 21 days, suffering from cryptosporidiosis were examined. It is recommended to perform the histopathological identification of cryptosporidia by prolongation of the basic staining with haematoxylin to one hour and by additional staining with tartrazine. Out of the special methods the most suitable are as follows: Wolbach's modification of Giems' staining method or staining with toluidine or polychromous blue. Mucopolysaccharides are in the granules and in the capsules of some developmental stages of cryptosporidia. By means of scanning electron microscope (SEM) cryptosporidia are clearly visible. As a rule, they are more or less immersed in the microvilli of enterocytes. In the intestines destructed by inflammation they occur even outside the epithelium, in the cases studied in the necrotic matter obstructing the outlets of the crypts of Lieberkühn in the caecum of the calves.     

Gill oxidative stress and histopathological biomarkers of pollution impacts in Nile tilapia from Lake Mariut and Lake Edku, Egypt

Various oxidative stress and histopathological biomarkers in gill tissues of Nile tilapia Oreochromis niloticus were investigated. Fish were collected from four sites that differ in their extent of pollution load, including heavy metals: the southeast basin (SEB), main basin (MB), and northwest basin (NWB) of Lake Mariut; and Boughaz El-Maadiya, a channel in Lake Edku. The oxidative stress biomarkers that were analyzed included lipid peroxidation (LPO), superoxide dismutase (SOD), catalase (CAT), and glutathione redox cycle enzymes (glutathione peroxidase [GPx] and glutathione reductase [GR]). Levels of reduced glutathione (GSH) were also evaluated. Gill morphology was analyzed by light microscopy and scanning electron microscopy (SEM). Gill LPO was significantly higher in gill tissues of fish collected from the more heavily contaminated MB (40.0%) and NWB (51.4%) sites than in gill samples from the less-contaminated (reference) site, the SEB. Gill LPO in fish from Lake Edku was intermediate but was not significantly higher (17.1%) than the reference. The activities of antioxidant enzymes and the redox-sensitive thiol compound GSH were significantly lower in gill samples from the disturbed sites than in samples from the reference site. Specifically, SOD in MB, NWB, and Lake Edku samples; CAT and GPx in NWB samples; and GR activity and GSH content in MB and NWB samples were lower than those in SEB samples. In most cases, gill tissues from Lake Edku fish had intermediate levels of antioxidants. The main histopathological alterations observed in gills were epithelial lifting, hyperplasia and hypertrophy of the respiratory epithelium, lamellar fusion, and aneurysms. In addition, SEM results demonstrated transformation of the surface structure of epithelial pavement cells. Pathological reactions in the gills of Nile tilapia were most severe at the MB and NWB sites. Our findings suggest that Nile tilapia responded differently according to the environmental stress index in each sampling area. This study is the first to report gill oxidative stress and histopathologies in Nile tilapia from Egyptian aquatic environments.