Lamellar Hyperplasia Research Articles

Acute exposure of zebrafish (Danio rerio) adults to psychotria carthagenensis leaf extracts: chemical profile, lack of genotoxicity and histological changes

Psychotria carthagenensis is a shrubby plant, often consumed by traditional populations in religious rituals. Previous studies have shown that this plant’s infusion can inhibit the activity of Acetylcholinesterase (AChE) in rats. Despite the therapeutic potential, there is a lack of research regarding its possible toxicological and genotoxic effects. Hence, this study aimed to analyze the chemical profile of the ethanol extract from P. carthagenensis leaves by LC-DAD-MS and assess its possible toxicity and genotoxicity in zebrafish (Danio rerio). Adult zebrafish (N = 9/group) were exposed at different concentrations and the LC50 was calculated. Frequencies of micronucleus (MN) and nuclear abnormalities (NA) were estimated for genotoxic effects, and degree of tissue changes (DTC) was used to assess the liver and gill histopathology. From the LC-DAD-MS analyses, the identified compounds included N-fructosyl valine, ethyl hexoside, 5-O-E-caffeoylquinic acid, N-feruloylagmatime, roseoside, di-O-deoxyhexoyl-hexosyl quercetin, loiolide, and oleamide. The calculated values of LC50 did not vary significantly during the time of exposure. At the concentrations of 1.25, 2.5, 3.75, 5, 7.5, 10 and 15 mg/L, there was no genotoxicity, and only low to moderate toxicity for the tissues was observed, despite mortality of 100% at doses of 20–100 mg/L of P. carthagenensis ethanolic leaf extract. There were changes in cytoplasm of hepatocytes at 1.25 mg/L, and karyorrhexis, karyolysis and megalocytosis at 10 mg/L. In the gills, the alterations were primary lamellar hyperplasia in all concentrations, and at 10 mg/L, secondary lamellar edema and vascular hyperemia were common. Additionally, the chemical composition of P. carthagenensis was expanded.

Salinity induced changes in growth and gill structure of freshwater carp, Cyprinus carpio Linn.

Aim: The present study was carried out to evaluate salinity induced changes in the growth and gill structure of cosmopolitan freshwater carp, Cyprinus carpio (Linn.). Methodology: Inland saline water (15 ‰ or g l-1) was collected from the salt affected waterlogged areas of village Shajrana (30.3346 0N, 74.1196 0E) in District Fazilka, Punjab (India) and diluted with underground freshwater for preparing different salinity levels (2 to 10 ‰). Fingerlings (10 ±2 cm) of C. carpio were exposed to 0 (control), 2, 4, 6, 8 and 10 ‰ salinity after gradual acclimatization with salinity increase @ 1 ‰ hr-1. Results: At the end of 120 days of rearing under saline conditions, 100% fish survival was observed up to 6 ‰ salinity, while 86.66 and 70.00% survival was recorded in 8 and 10 ‰, respectively. However, fish growth declined significantly at all salinity levels (p≤0.05). Gill structure was also affected at all salinity level, but pronounced changes were observed at salinity levels ≥ 6 ‰, including lamellar oedema, epithelial lifting, lamellar hyperplasia, hypertrophy, lamellar fusion, hyalinisation, aneurysm, blood congestion, etc. Interpretation: The results offer referral database to explore optimised economic production of common carp in inland saline areas of the region at salinity levels below 6 ‰. Key words: Carp, Gills, Growth, Production, Salinity, Survival

Fish Gills as Morphological Indicators of Exposure to Four Environmentally Relevant Pharmaceuticals

Pharmaceuticals are a fast-emerging class of environmentally relevant contaminants of concern, due to their ubiquitous presence in surfaces waters consequent on human and veterinary use. Their relatively unknown effects on non-target organisms and probable deleterious effects on ecosystem health and complexity make it necessary to develop the use of simple early warning detection systems to validate their probable presences in the aquatic environmental niche.. In this paper, we present the results of the effects on the gills of a local farmed catfish species, Clarias gariepinus of four pharmaceuticals detected in the Lagos lagoon at concentrations exceeding determined no observed effect concentrations (noec) in literature. Histopathological changes observed included lamellar hyperplasia, hypertrophy, epithelial lifting, necrosis and atrophy. This study validates the continuing histological utility of fish gills as sensitive indicators of exposure to xenobiotics, and the inherent capacity of pharmaceutical residues in water to elicit detectable pathologies in various fish organs, even at extremely low concentrations.

Acute toxicity of trichlorfon and histological changes in the gills of Arapaima gigas, a neotropical fish from Amazon

Trichlorfon is a widely used drug to control ectoparasites in fish farming and is effective in treating protozoa, helminths, and crustaceans. However, prior to a concentration recommendation made for therapeutic purposes in aquaculture, it is necessary to establish its toxicity, behavioural and histological effects, and determine a safe use dose for each target species. Here we investigated the acute toxicity (LC50-12 h) of trichlorfon and its behavioural and histological effects on arapaima (Arapaima gigas) larvae and juveniles. For the acute toxicity tests (LC50), arapaima larvae and juveniles were exposed at 0, 50, 250, 500, 750, 1.000 and 1.250 mg L−1 of trichlorfon for 12 h. During this period, behavioural changes, mortality, and the water physico-chemical parameters were recorded. Immediately after a toxicity test, gills were collected for the histological analysis. The LC50–12 h for the arapaima larvae and juveniles were 411.8 mg L−1 and 587.3 mg L−1, respectively. The fish exposed for different times to the 250, 500, 750, 1.000 and 1.250 mg L−1 concentrations showed a dose-dependent effect with behavioural changes, such as erratic swimming, loss of balance, lethargy, gasping at the water surface and muscle spasms. Histological changes, such as lamellar hyperplasia and cell hypertrophy, total lamellar fusion, dilation, and vascular congestion in the central filament vessel, interlamellar hyperplasia, capillary congestion, proliferation of mucous cells, dilation of the venous sinus and capillaries, leukocyte infiltrate and haemorrhage, were observed and ranged from mild to diffuse lesions. However, branchial necrosis was presented only in the fish exposed to ≥ 500 mg L−1 of trichlorfon. In conclusion, sublethal trichlorfon concentrations (≤ 250 mg L−1) and short exposure times (up to 180 min) can be safely used in arapaima larvae and juveniles because they do not alter their normal behaviour nor induce mortalities and/or irreversible injuries to their gills.

Effects of trichlorfon organophosphate on the morphology of the gills and liver of Pseudoplatystoma corruscans

Possible changes in the morphology of the gills and liver of P. corruscans after exposure to TCF were evaluated. The fish were distributed into five groups in triplicate (n = 10 in each group: 0.0; 0.125; 0.25; 0.5; 1.0 mg/L of the commercial product NEGUVON®, for 96 h. TCF induced a high histopathological index associated with circulatory disorders (congestion and aneurysm), as well as progressive changes (lamellar hyperplasia, capillary dilation, epithelial detachment and edema) at the highest concentrations (0.25; 0.5 and 1.0 mg/L). In addition to the histopathological disturbances, there was an increase in the levels of neutral glycoproteins and alterations in the histomorphometry of the secondary lamellae (total height, apical region, median, basal and interlamellar) at the highest concentrations. In the liver, the alterations were more intensified, in addition to circulatory (congestion) and regressive disturbances (loss of hepatocyte architecture and accumulation of intracellular substances), progressive alterations (focal necrosis) were observed, indicating a high degree of hepatic tissue involvement. The results indicate that the use of TCF concentrations above 0.25 mg/L promotes histopathological, histomorphometric and histochemical changes in the gills and liver of P. corruscans, imposing a biological risk that affects the aquatic environment and the health of fish.

Study of parasitic effect of Argulus sp. on Indian major carps in pond culture

The current investigation was undertaken to study the parasitic effects of Argulus sp. on Indian major carps. Only symptomatic and moribund samples of diseased fishes comprised of fingerlings and adults of Labeo rohita, Catla catla and Cirrhinus mrigala were collected for experiments. The affected fishes showed weight loss, retarded growth, restlessness, erratic swimming behaviour and loss of appetite. The skin showed abnormally pigmented while gills, kidney, liver and spleen of the affected fishes were pale in colour. The most significant histopathological lesions in the gills were the secondary lamellar hyperplasia, haemorrhages in the tips of the primary lamellae and fusion of both secondary and primary gill lamellae. Skin showed epidermis was lost but the epidermal cells in the vicinity of the sites of attachment were hyperplastic, muscular layer showed degenerative and necrotic. Kidney showed multifocal enlargement of glomeruli, glomerular tufts in many places shrunken, fragmented and necrosed, in renal tubules vacuolar degeneration. Fingerlings and adults of rohu were found highly susceptible to this disease than other IMC.

Parasitic Lesions in Fish in the Federal District, Brazil

Background: Brazil has a vast territory and favorable climatic conditions that allow the cultivation of freshwater fish. The intensification of the productive system can cause an imbalance in the aquatic environment as a result of poor water quality, nutritional deficiencies and infectious or parasitic diseases. The laboratory diagnosis and the determination of the prevalence of the main lesions, which occur in a certain region, help to guide towards the etiological diagnosis. This study aimed to describe the main parasitic lesions in fish in the routine at the Veterinary Pathology Laboratory of the Universidade de Brasília. Material, Methods & Results: All records of fish with parasitic lesions were recovered. Those cases in which there was an intralesional parasite and which presented lesions compatible with the parasite were included. The screening of ectoparasites was done by scraping the superficial mucus from the gills and skin. Organ sections were routinely processed for histopathologyand stained with hematoxylin-eosin (HE). In some records, parasitological identification was carried. The information was divided into the species of the affected fish, epidemiology of the outbreak (water quality, temperature, type of breeding), lesion distribution, etiology and macroscopic and microscopic changes. The resulting data was organized in absolute frequency and percentage. In this study, 22 cases were counted, between individual deaths and outbreaks, totaling 83 necropsied teleost fish.Inflammatory changes of parasitic origin were seen in 13/22 (59%) of the cases had lesions of parasitic origin. Skin lesions and gills were the most relevant. Macroscopically, red areas or spots of hyperemia or hemorrhage on the body surface were the most prevalent findings. Under microscopy, proliferative gill inflammation was the most relevant diagnosis. Pscinoodinium pilullare (Dinoflagellida), Ichthyophthirius multifiliis (Ciliophora), and monogenetic worms (Monogenea) were the main parasites found. Trichodina sp. (Ciliophora), Ichtyobodo sp. (Kinetoplastida), Amoebas, and Chilodonella sp. (Ciliophora), were seen in fewer numbers. An unusual case suggestive of parasitism by Eustrongylides sp. (Nematoda), in a pirá-brasília (Simpsonichthys boitonei), specimen has been recorded.Discussion: The diagnoses were based on epidemiology, anatomopathological and parasitological findings. The most frequent and significantly lethal lesion in the study was proliferative and / or hyperplastic branchitis. Proliferative branchitis with lamellar epithelial hyperplasia (LEH) is a response to some type of chemical or mechanical injury to the gill epithelium in order to protect the capillaries from further damage or microbial penetration. However, it also increases the diffusion distance between capillaries and the environment and, therefore, hinders breathing, excretory and osmoregulatory functions. Protozoan infections and monogenetic worms in general generated LEH and skin lesions of mechanical origin. Secondary bacterial infection, were observed in this parasitosis determining the cause of death of the fish. Its pathogenicity comes from the lesions caused by the colonization and histophagy of the epithelial surfaces, mainly gills and skin, causing epithelial proliferation, lamellar cell fusion, epithelial cell degeneration and necrosis forming several ulcers in the epithelium after the release of mature trophies. The pathogenesis of parasitism by Eustrongylides spp. is considerable when there is a large quantity of these larvae that can cause intestinal obstruction, rupture and compression of viscera, of greater importance in small fish. The main parasites of necropsied fish were protozoa and monogenetic worms, which mainly cause branquitis and dermatitis in varying grades.

Essential oil of Lippia grata (Verbenaceae) is effective in the control of monogenean infections in Colossoma macropomum gills, a large Serrasalmidae fish from Amazon

This study investigated the efficacy of Lippia grata essential oil (EO) against monogeneans of Colossoma macropomum and effects on the haematology after immersion baths. In the in vitro assays, the efficacy of 100, 250, 350 and 700 mg/L of L. grata EO and two controls were tested, of which one control was with the cultivation tank water and the other was with the tank water and 70% ethyl alcohol. Composition of majority bioactive compounds in the EO was carvacrol (48.12%), p-cymene (24.39%) and γ-terpinene (2.49%). Anacanthorus spathulatus, Notozothecium janauachensis, Mymarothecium boegeri and Linguadactyloides brinkmanni obtained of fish gills showed 100% immobilization in the in vitro assays when exposed to 700 and 350 mg/L after 30 min and 2 hr respectively. Fish showed tolerance to 700 mg/L of the EO, which was used in therapeutic in three consecutive baths, which caused lamellar hyperplasia in C. macropomum gills. The 700 mg/L of the EO in the therapeutic baths showed an efficacy of 95.1% against the monogeneans and caused decrease in haemoglobin, whereas control fish showed a reduction in mean corpuscular haemoglobin concentration. Results indicated that 700 mg/L of L. grata EO is a safe and effective concentration to treat C. macropomum infected with monogeneans.

Histopathological alterations in gills of a fish (Astyanax bifasciatus) in neotropical streams: negative effects of riparian forest reduction and presence of pesticides.

The reduction of riparian vegetation around aquatic environments causes several physicochemical alterations and favors the entry of pesticides via surface runoff. Such changes have negative effects on aquatic organisms. In this study, we evaluated histopathological alterations in gills of Astyanax bifasciatus to test the hypothesis that more severe histopathological alterations occur in gills of fish from streams with higher agricultural impact from the surrounding area. The specimens were collected by electrofishing in seven streams of the lower Iguaçu basin between August 2015 and February 2016. The gills were processed according to routine histological methods and examined by light microscopy. The histopathological alterations, mainly stage II (lamellar aneurysm and total fusion of lamellae), were observed in fish collected in streams with higher agricultural activity. In these streams, the histopathological index indicated slight to moderate organ lesions. In contrast, in streams with more vegetation cover, fish collected presented stage I histopathological alterations (lamellar edema and lamellar hyperplasia), and the HI indicated normal functioning of the gills. In addition, chloride and acid mucous cells were more abundant in the gills of fish collected in rural streams. Our findings demonstrate that more severe histopathological alterations were registered in fish collected from streams with intense agricultural activity in the surrounding area. Therefore, it highlights that vegetation cover around the streams is a positive force for the conservation and health of aquatic organisms.

Effect of the dietary linoleic/α-linolenic ratio (n6/n3) on histopathological alterations caused by suboptimal temperature in tilapia (Oreochromis niloticus).

Tilapias are produced worldwide, including subtropical areas. In turn, dietary fatty acids can enhance resistance under cold stress. The present study reports the nutritional effect on suboptimal temperature tolerance based on histopathological alterations in tilapia (Oreochromis niloticus). Fish (initial weight: 34.02 ± 0.79) were exposed to two different temperatures (20 °C and 30 °C) for 30 days. Under both conditions, fish were fed two different diets based on the linoleic/α-linolenic ratio (n6/n3 = 12.02 and n6/n3 = 3.85). The most important alterations in liver caused by temperature included high cellular hyperplasia in fish at 30 °C (P < 0.05). Suboptimal temperature also caused gills aneurysm, lamellar lifting and low hyperplasia (P < 0.05). Cytoplasmic vacuolization decreased and nuclear displacement increased in the fish fed n6/n3 = 3.85 (P < 0.05). Suboptimal temperature provided high gills aneurysm, epithelial lamellae lifting and low lamellar hyperplasia (P < 0.05). Conversely, the fatty acid content (n6/n3 = 3.85) in the diet provided low lamellar lifting and fusions (P < 0.05). Finally, tilapia showed an increased glycogen storage trend in gills and muscle at low temperature (P < 0.05). Our study suggests that when feeding tilapia a high dietary n3 content, this fish is able to respond to suboptimal temperature in subtropical aquaculture facilities.

Chronic effects of copper and zinc on the fish, Etroplus suratensis (Bloch, 1790) by continuous flow through (CFT) bioassay

Copper (Cu) and zinc (Zn) play a vital role in the growth and development, however increased uptake causes deleterious effects in normal functioning of organisms. We have demonstrated in this contribution the tolerance limit of Cu and Zn on Etroplus suratensis (pearl spot) by Continuous Flow Through (CFT) bioassay and the biomarker responses. The accumulation for Cu, Zn and selected trace metals (Cr, Cd, Ni and Pb) from field conditions, as well Geo-accumulation index (I geo) and Contamination factor (C.F) suggested moderate to heavy pollution in the Cochin estuarine system. The 96 h LC50 values for Cu was 1.74 ± 0.04 mg L−1and that for Zn was 24.36 ± 0.58 mg L−1 at 95% confidence interval. No observed effect concentration (NOEC) and low observed effect concentration (LOEC) for Cu and Zn were derived based on the survival rates. Chronic toxicity values for Cu and Zn were 0.23 mg L−1 and 2.005 mg L−1 respectively for 30 days period. The histological, biochemical, hematological and behavioral parameters showed significant variations at sublethal concentrations. Lamellar hyperplasia in gills, vacuolation combined with necrosis in liver, increased occurrence of melanomacrophage centres in spleen were noticed at chronic levels for both Cu and Zn. Tissue specific bioconcentration was observed for zinc and copper in gill and liver respectively, with least rate of bioconcentration observed in muscle tissues. Malaonate Dehydrogenase (MDH), Super oxide dismutase (SOD), Nonspecific esterase (EST) activity significantly varied compared to control at NOEC and LOEC values in both the metals. The hematological and genotoxic alterations as decrease in erythrocyte count, lymphocytes, hemoglobin concentration and hematocrit percentage were significantly reduced (p < 0.05) and increased thrombocytes and neutrophils, increased frequency of micronuclei, lobed, blebbed and notched nuclei and binucleate cells were characteristic for the metals at the sublethal concentrations. The frequency of behavioral changes remained significantly higher at chronic level than the control group. Thus such CFT based studies are important for precisely mapping the toxicity changes in organisms and also to develop suitable water quality guidelines.

Histomorphometric changes in the gill of Clarias gariepinus exposed to acute concentrations of chlorpyrifos

Abstract Introduction: Chlorpyrifos is an agrochemical pesticide of fame that is commonly used by farmers to control pests. This chemical may become harmful to fish when washed into waterways leading to aquatic environment. This study investigates the histo-morphometric changes in the gills of African Catfish Clarias gariepinus exposed to acute toxic concentrations of chlorpyrifos. Materials and Methods: Clarias gariepinus fingerlings (weight 7.28 ± 0.03g and length 4.82 ± 0.06cm) were exposed to grades of (6.25, 12.5, 25, 50 and 100 pml/L) chlorpyrifos in solution. The 96 hour LC50 value of chlorpyrifos was found to be 160 pml/L. Results: The grades of chlorpyrifos induced several gill histo-architectural damages such as: moderate to severe gill epithelia sloughing, primary and secondary lamellar hyperplasia and central veinous congestion in the parenchyma with pronounced severity in fish exposed to higher concentrations. Similarly, the gill morphometrics (secondary lamellar length, width, interlamellar distance and surface area) were markedly altered by the graded concentrations of chlorpyrifos. Conclusion: Chlorpyrifos appears to be very toxic to ish and therefore, its use near ish farms or in areas close to aquatic environments should be discouraged.

Infectious bacterial pathogens, parasites and pathological correlations of sewage pollution as an important threat to farmed fishes in Egypt

This paper is a part of a multi-disciplinary research “Application of Decentralized On-Site Water Treatment System in Egypt for Use in Agriculture and Producing Safe Fish and Animal Proteins”.The project aimed to investigate the environmental impact of implementing sewage water before and after treatment using the effluent of the on-site decentralized Japanese' Johkasou system, in agriculture and producing fish protein. The aim is to establish such system in Egypt to strengthen the sanitary conditions of water resources. In the present study, the impact of the sewage pollution in some fish farms at El-Fayyum, Port Said and El-Dakahlia governorates in Egypt was carried out. Water and fish (Oreochromis niloticus and Mugil cephalus) samples were collected from private fish farms of such localities. Bacteriological and chemical examination of water samples revealed the existence of coliforms and many other bacterial species of significant human health hazards. The chemical parameters of water showed a marked deviation from normal levels while examination of fish flesh specimens indicated contamination with Streptococcus Sp., Staphylococcus Sp., and Salmonella in all examined localities. Other bacterial isolates of human health importance (Morganella morganii, Pseudomonas cepacia and Enterococcos durans) were identified. The parasitological examination revealed the presence of encysted metacercariae (EMC); Diplostomatidae, Prohemistomatidae and Heterphyidae. Moreover, two protozoan parasites (Mxyoboulus tilapiae and Ichthyophthirius multifilis) were also recorded. The histopathological examination revealed mild tissue reaction in case of bacterial infection and severe pathological lesions in different organs in case of EMC infection. Lamellar hyperplasia and mononuclear cell infiltration in branchial tissue was common findings. In skeletal muscles, atrophy of muscle fibres, myolysis and myophagia were detected.

Partial substitution, with their chelated complexes, of the inorganic zinc, copper and manganese in sow diets reduced the laminitic lesions in the claws and improved the morphometric characteristics of the hoof horn of sows from three Greek herds.

BackgroundHoof lesions in sows have been associated with lameness and poor hoof horn quality. The mechanical strength and quality of hoof horn is determined by the density and diameter of horn tubules, which were recently associated with the severity of lesions on the hoof wall of sows. Histologic changes that have previously been described in cases of bovine laminitis, have also been observed in the dermis and epidermis of the sows’ claws. Trace elements, particularly zinc, copper and manganese, occupy important roles as enzyme catalysts in the process of keratin synthesis which determines the quality and the integrity of the hoof epidermis. Therefore, the objective of this study was to investigate the effect of diet supplementation with chelated zinc, copper and manganese, partially substituting their inorganic form, on sow claw health and hoof horn quality assessed by macroscopic, histologic and morphometric examination.ResultsClinically, the total claw lesion score was significantly lower in claws of sows which received the “organic” diet compared to those of sows on the “inorganic” diet. Histologically, lamellar hyperplasia was the most frequently recorded change in the epidermis of the sows’ claws regardless of the diet’s mineral source. The claws of the sows which received the organic diet were more likely to have none or less histologic changes than at least one or more, respectively, compared to those of the sows on the “inorganic” diet. Morphometrically, the density and vertical and horizontal diameters of the horn tubules was significantly higher and smaller, respectively, in the hoof horn of sows which received the “organic” compared to those which received the “inorganic” source diet.ConclusionsPartial substitution of the inorganic zinc, copper and manganese in sows’ diet with their chelated complexes, provided a comparative advantage against a conventional, inorganic mineral source diet, at least under the conditions examined in the current study, in terms of macroscopic, histologic and morphometric criteria, characterizing the health and horn quality status of sows’ hooves.

Pathological evaluation of claw lesions I culled sows from a Greek herd

Objectives: To characterize macroscopic claw lesions of culled sows, describe the histologic characteristics observed in the laminar corium and investigate their associations with lesion severity, and compare the morphometric characteristics of horn tubules among claws according to lesion severity. Materials and methods: One front and the opposite rear foot from 74 culled sows of one herd were examined for lesions. From each claw, a tissue sample consisting of dermis and epidermis was examined histologically for changes suggesting laminitis. Slices from the lateral claws of the rear feet of 48 sows were examined morphometrically to evaluate the density and size of horn tubules. Results: The most frequent lesions were those located on the heel, wall, and white line, with 146 (49.3%), 94 (31.8%), and 81 (27.4%) affected claws, respectively, among the 296 examined. Lamellar hyperplasia was the most frequently recorded characteristic in the epidermis of 87 of 296 claws (29.4%) in 51 of the 74 examined sows (68.9%). The total lesion score of the claw was higher (P &lt; .001) when lamellar hyperplasia was recorded than when no histologic change was recorded. The density of horn tubules was lowest (P = .018) and the size was largest (P &lt; .001) among animals with severe wall lesions, compared to those without wall lesions. Implications: The histologic changes observed in the dermis and epidermis of the sows’ claws have been described in cases of equine and bovine laminitis. Sow laminitis may frequently occur, causing production of low quality hoof horn.

Infantile Cortical Hyperostosis: Report of a Case with Observations on Clinical Manifestations, Radiology, and Pathology with a Late Follow-Up of Eight Years.

Purpose. The purpose of our study was to investigate clinical manifestations, roentgen images, histopathological studies, and evolution of the disease in patient displaying infantile cortical hyperostosis. Methods. Roentgenograms were made to evaluate a neonatal patient presenting multiple soft-tissue swellings. The initial radiographs insinuated that the disease had been present for some time in utero. Bone puncture biopsy of the tibia for histopathological observation and diagnosis conclusions was performed. Results. The disease was demonstrated radiographically by massive cortical diaphyseal thickening and also extensive periosteal new bone formation surrounding several bones. Results in blood count were as follows: discrete anemia, moderate leukocytosis, and elevated sedimentation rate. Histological pattern of tissue removed from tibia showed lamellar cortical bones and hyperplasia. Biopsy studies disclosed no evidence of neoplasia as well as of bacterial infection. Comments. Clinical manifestations in a neonatal patient displaying infantile cortical hyperostosis have gradually decreased. Radiograph findings have demonstrated complete recovery of bones manifested by the disease. The pathologic findings are in accordance with previous microscopic examination summarized by the literature. Total patient cure, without sequels, could be demonstrated.

Municipal wastewater-induced histopathological lesions in gills of freshwater fish Labeo rohita – A SEM study

The fingerlings of fish Labeo rohita were subjected to chronic (15, 30, and 60 days) and recovery (60 days) experiments for histopathological studies using sublethal concentrations (35.4%, 26.6%, and 17.7% v/v) of municipal wastewater of Tung Dhab drain, Amritsar, India. Control fish were maintained for same exposure periods using tap water. Scanning electron microscopic studies exhibited dose- and duration-dependent alterations on gill surface ultra-structure. Control fish exhibited functionally normal gills with normal mean degree of tissue change. Fifteen different types of lesions were identified out of which 9, 5, and 1 were of the stage I, II, and III, respectively. The mean degree of change values in the treated groups ranged from 14.8 ± 0.5 (17.7%, 15 days) to 116. ± 17 (35.4%, 60 days) demonstrated gill damages of slight to irreparable lesions. The recorded alterations were epithelial lifting and intraepithelial edema, reduction of water spaces, lamellar fusion, lamellar telangiectasia, swelling and fusion of filaments, sloughing of lamellar epithelium, hypertrophy and hyperplasia of mucous and chloride cells, and necrotic lamellae. The recovery experiments showed positive improvement as the intensity of damage was less than recorded for 60 days of treatment. This study clearly indicated toxicity of drain water to fish at cellular and organ level. The study recommends environmental agencies to make and enforce decisions regarding improvement of water quality of Tung Dhab drain, and further incorporating histopathological studies as an analytical tool.

Chemical and histological comparisons between Brevoortia sp. (menhaden) collected in fall 2010 from Barataria Bay, LA and Delaware Bay, NJ following the DeepWater Horizon (DWH) oil spill

Body burdens of PAHs were compared to histological effects in menhaden (Family: Clupeidae, Genus: Brevoortia) collected in fall 2010 from Barataria Bay, LA (BBLA) and Delaware Bay, NJ (DBNJ). Barataria Bay was heavily oiled during the DeepWater Horizon (DWH) oil spill, while Delaware Bay although urbanized had no reported recent oil spills. GCMS analyses of pre-spill 2009, BBLA and DBNJ fish found predominantly C2/C3 phenanthrene (1.28–6.52 ng/mg). However, BBLA also contained five higher molecular weight PAHs (0.06–0.34 ng/mg DW). Fluorescent aromatic compound spectroscopy (FACS) of gastrointestinal (GI) tract tissue showed statistically higher levels of hydroxypyrene-like PAHs in DBNJ than BBLA fish. Histopathologic lesions were more prevalent in BBLA than DBNJ fish. The lesion prevalence (gill, trunk kidney, epidermis, stomach) in the BBLA menhaden were significantly higher and more severe than observed in the DBNJ menhaden. Reversible lesions included gill lamellar hyperplasia, adhesions, edema, and epidermal hyperplasia. The increased pigmented macrophage centers were indicative of activated macrophages responding to connective tissue damage or other antigens. The liver hepatic necrosis and renal tissue mineralization may well have undergone repair, but damage to the kidney nephrons and hepatic/biliary regions of the liver would be slower to resolve and apparently remained after elimination of PAHs. Therefore, a direct cause and effect between DWH oil spill and increased lesion prevalence in BBLA menhaden could not be established.

Epitheliocystis in lake trout Salvelinus namaycush (Walbaum) is associated with a β‐proteobacteria

Lake trout Salvelinus namaycush (Walbaum) raised for stocking experienced yearly (2011-13) winter epizootics of epitheliocystis. Affected fish were dispersed on the bottom of the tank, had decreased feed and fright response, and mortality often reached 40%. Peak mortality occurred within 3weeks of the appearance of clinical signs, and outbreaks typically lasted 6weeks. Affected fish had no gross lesions but histologically had branchial epithelial necrosis and lamellar hyperplasia, with small to large numbers of scattered epithelial cells containing 10- to 20-μm inclusions. A longitudinal study was undertaken of one annual outbreak, and lamellar hyperplasia was most closely associated with mortality. The number of inclusions was statistically greater (P<0.05) before and during peak mortality, but inclusions were present in low numbers before clinical signs occurred. Results of histochemical staining, immunohistochemistry and transmission electron microscopy supported the presence of a β-proteobacteria rather than a Chlamydiales bacterium within inclusions. PCR primers to identify Chlamydiales did not give consistent results. However, the use of universal 16S rDNA bacterial primers in conjunction with laser capture microdissection of inclusions demonstrated that a β-proteobacteria was consistently associated with affected gills and is more likely the cause of the disease in lake trout.

Sex-response differences of immunological and histopathological biomarkers in gill of Prochilodus argenteus from a polluted river in southeast Brazil

The fish gill is in direct and standing contact with the immediate external environment and, therefore, is highly vulnerable to aquatic pollutants. In this study, Prochilodus argenteus were caught at two different points in São Francisco river. The first point is located near Três Marias dam, while the second is placed downstream the Abaeté river. Chemical approaches showed the presence of metals contamination in the first point. Thus, the main goal of this study was to investigate the possible toxic effects of these contaminants and the likely use of biomarkers on fish gills. Biometric data of length and weight of fish were obtained in order to calculate the condition factor as an organismal biomarker. The histological changes in gills and alterations in mucous and rodlet cells occurrence were detected microscopically and evaluated with quantitative analyses. Myeloperoxidase (MPO) and Eosinophil Peroxidase (EPO) were also assessed in fish gill. The analysis of the water and sediment samples revealed the presence of metals at the two points. As and Cd were detected at higher concentrations at point 1. The presence of lamellar cell hyperplasia, lamellar fusion, lamellar edema and inflammatory foci varied according to the point. Additionally, mucous and rodlet cells and MPO and EPO activities showed variability according to the environmental conditions. Furthermore, with exception of lamellar hyperplasia and eosinophil peroxidase activity, all others parameters showed sex-variation responses. At the first point, male fish showed a chronical inflammation in gills due to the lowest activity of MPO and EPO, as well as low occurrence of inflammatory foci and glycoprotein secretion by mucous cells, while female fish presented an opposite pattern of response to the same environmental conditions. Therefore, we suggest the use of such biomarkers in future monitoring of aquatic systems, taking into account the sex-variation responses.