Probiotic supplementation affects IGF-1 and leptin levels in Nile tilapia hepatopancreatic tissue / Suplementação com probiótico afeta os níveis de IGF-1 e leptina no tecido hepatopancreático de tilápias-do-nilo

This work aimed to assess the effect of the probiotic strain, Lactobacillus plantarum, on the levels of leptin, IGF-1 and their receptors on the hepatopancreatic tissues of Nile tilapia (Oreochromis niloticus) and then correlate fish growth performance and gut microbiological parameters. Fish juveniles (±23g) were reared in a recirculation system with constant aeration and temperature (25°C). They were distributed into six polyethylene tanks (45L) and fed twice a day at 5% of the tank biomass with the respective diets: control (commercial diet without probiotic) and supplemented with L. plantarum inoculum (1 x 108 CFU mL-1), both in triplicate. After 30 days of feeding, L. plantarum-fed fishes showed greater weekly growth rate, final weight, and feed conversion rate, in addition to higher count of lactic-acid bacteria and lower count of pathogenic bacteria in the intestinal tract, when compared to the control group. The immunostaining intensity for IGF-1 and leptin hormones was lower after L. plantarum supplementation than in the control group, with no change in the level for receptors. This reduction could implicate important changes in fish metabolism and homeostasis.(AU)

O presente trabalho teve como objetivo avaliar o efeito da cepa probiótica Lactobacillus plantarum sobre os níveis de leptina, IGF-1 e seus receptores no tecido hepatopancreático de tilápia-do-nilo (Oreochromis niloticus) e correlacionar com o desempenho zootécnico e os parâmetros microbiológicos intestinais dos peixes. Juvenis de tilápia-do-nilo (±23g) foram distribuídos em seis tanques de polietileno (45L) conectados a um sistema de recirculação, com aeração e temperatura constantes (25°C). Os peixes foram alimentados duas vezes ao dia, a 5% da biomassa do tanque, com as respectivas dietas: controle (dieta comercial sem probiótico) e suplementada com L. plantarum (1 x 108 UFC mL-1), ambas em triplicata. Após 30 dias de cultivo, os peixes alimentados com L. plantarum apresentaram maiores ganho de peso semanal, peso final e conversão alimentar, bem como maior contagem de bactérias ácido-láticas e menor contagem de bactérias patogênicas no trato intestinal das tilápias alimentadas com dieta probiótica, em comparação ao grupo controle. A intensidade da imunomarcação para os hormônios IGF-1 e leptina foi menor com a suplementação de L. plantarum do que no grupo controle, sem alterar os níveis de seus receptores. Essa redução pode implicar mudanças importantes no metabolismo e na homeostase dos peixes.(AU)

Histomorphologic characterization of the hepatopancreas of freshwater prawn Macrobrachium rosenbergii (De Man, 1879) / Caracterização histomorfológica do hepatopâncreas do camarão-de-água-doce Macrobrachium rosenbergii (De Man, 1879)

This study aimed to describe the Macrobrachium rosenbergii hepatopancreas histomorphology. The hepatopancreas is constituted by a set of blind end tubules, divided into proximal, middle, and distal regions, with the epithelium formed by E, F, B, R, and M cells differently of other species. Measurements of the length and width of the tubules were 419.64+69.09µm and 117.42+16.99µm, respectively. The percentage of each cell type per region was: proximal region (40%B, 20%F, 6.7%M, 33.3%R), middle region (45.4%B, 18.2%F, 9.1%M, and 27.3%R) and distal region (36.4%E, 27.2%B, 18.2%F, 9.1%M, 9.1%R). Cell B that stores glycogen and lipids, is the most commonly found cell in proximal and middle regions. In the distal region, the E, responsible for the mitosis, is the most prominent. M, responsible by nutrient storage, is numerically constant among the portions differently in the Macrobrachium amazonicum. The study for the first time also suggests that in addition to digestive enzymes, the F cell produces protective mucus. The present study generated for the first time a morphometric profile of M. rosenbergii hepatopancreas, demonstrating differences from other species, and can be an important tool for new studies in nutrition, reproduction, and production with the species.(AU)

O objetivo do presente estudo foi descrever a histomorfologia do hepatopâncreas do camarão-de-água-doce Macrobrachium rosenbergii. Observou-se que ele é constituído por um conjunto de túbulos de fundo cego, sendo cada túbulo dividido em regiões proximal, média e distal, com o epitélio formado por cinco tipos de células (E, F, B, R, M), diferentemente de outras espécies. As medidas de comprimento e largura dos túbulos foram de 419,64+69,09µm e 117,42+16,99µm, respectivamente. A porcentagem de cada tipo celular por região foi: região proximal (40%B, 20%F, 6,7%M, 33,3%R), região média (45,4%B, 18,2%F, 9,1%M e 27,3%R) e região distal (36,4%E, 27,2%B, 18,2%F, 9,1%M, 9,1%R). Assim, a B, que armazena glicogênio e lipídeos, é a célula mais encontrada nas regiões proximal e média. Na região distal, a célula E, responsável pela mitose, é a mais encontrada. A célula M, responsável pelo acúmulo de nutrientes, tem um número constante de células nas porções do túbulo, diferentemente do Macrobrachium amazonicum. O estudo também sugere, pela primeira vez, que a célula F produz, além de enzimas digestivas, um muco protetor para o túbulo hepatopancreático. O presente estudo foi o primeiro a gerar um perfil morfométrico do hepatopâncreas do M. rosenbergii e demonstrou diferenças em relação a outras espécies, bem como serviu de importante ferramenta para novos estudos que abranjam a produção, a nutrição e a reprodução para a espécie.(AU)

Hypolobocera guayaquilensis (Decapoda: Pseudothelphusidae): A New Crab Intermediate Host of Paragonimus mexicanus in Manabí Province, Ecuador

To determine that Paragonimus sp. is actively transmitted in a tropical area of the Pacific region of Ecuador where human cases of pulmonary paragonimiasis have recently been documented, a total of 75 freshwater crabs were collected from 2 different streams in the Pedernales area of Manabí Province, Ecuador. All collected crabs were identified as Hypolobocera guayaquilensis based on morphological characteristics of the male gonopods. The hepatopancreas of each crab was examined by compressing it between 2 glass plates followed by observation under a stereomicroscope. Excysted Paragonimus metacercariae were detected in 39 (52.0%) crabs and their densities varied from 1 to 32 per infected crab. There was a positive relationship between crab size and metacercarial density. Sequences of the second internal transcribed spacer region of the ribosomal RNA gene of the Paragonimus metacercariae obtained in this study were identical to those of Paragonimus mexicanus deposited in the DDBJ/EMBL/GenBank database. Thus, the present study is the first to confirm that the crab species H. guayaquilensis is the second intermediate host of P. mexicanus in Manabí Province, Ecuador. Because this crab might be the possible source of human infections in this area, residents should pay attention to improper crab-eating habits related with a neglected parasitic disease, i.e., paragonimiasis.

Infections of Larval Stages of Dicrocoelium dendriticum and Brachylaima sp. in Brown Garden Snail, Helix aspersa, in Turkey

The aim of this study was to determine the presence and prevalence of larval stages of Dicrocoelium dendriticum and Brachylaima sp. in the first intermediate host, a species of land snail, Helix aspersa, in Turkey. A total of 211 snails were collected in April-May 2014 from pastures in Mersin District. Larval stages of D. dendriticum were identified under a light microscope. Hepatopancreas from naturally infected H. aspersa snails were examined histologically. The prevalence of larval stages of D. dendriticum and Brachylaima sp. in H. aspersa snails was found to be 2.4% and 1.9%, respectively, in Mersin, Turkey. Cercariae were not matured in sporocysts at the beginning of April; however, it was observed that cercariae matured and started to leave sporocysts by early-May. Thus, it was concluded that H. aspersa acts as an intermediate host to D. dendriticumin and Brachylaima sp. in Mersin, Turkey. A digenean trematode Brachylaima sp. was seen for the first time in Turkey.

Purification and characterization of 6-phosphogluconate dehydrogenase (6-PGD) from grass carp (Ctenopharyngodon idella) hepatopancreas.

6-Phosphogluconate dehydrogenase (6-PGD, E.C.: 1.1.1.44) was purified and characterized from the hepatopancreas of grass carp (Ctenopharyngodon idella) for the first time. Grass carp represents the second largest aquaculture industry in the world after silver carp, constituting 14.7% of the world aquaculture production, with an average annual increase of 14% in China, mainly as a source of food. The purification procedure involved a single 2’, 5’-ADP-Sepharose 4B affinity chromatographic step by using different elution buffers. The enzyme was purified 309-fold with a specific activity of 5.259 U/mg protein and yield of 68%. The purity and subunit molecular weights of the 6-PGD were checked on SDS-PAGE and purified enzyme showed a single band on the gel. The subunit molecular mass was 57 kDa, with an optimum pH, temperature and ionic strength at 7.96, 50oC and 100 mM Tris-HCl, respectively. The Km values of 6-PGA and NADP+ were 0.019 and 0.0052 mM, respectively, while Vm of 6-PGA and NADP+ was 0.69 U/ml. Dissociation constants (Ki) for 6-PGA and NADP+ were 2.05 and 0.12 mM, respectively. NADPH inhibited the enzyme in a competitive manner and its Ki value was 0.032 mM. The Cu2+, Zn2+, Cd2+ and Al3+ showed inhibitory effects on the enzyme with IC50 values of 0.293, 0.099, 0.045 and 1.526 mM, respectively. All tested metals inhibited the enzyme in a competitive manner, indicating that these metals might be toxic even at low concentrations for the 6-PGD. As the fish is one of valuable foodstuff of animal sources for human consumption, under certain environmental conditions, metal ions accumulated in fish up to a lethal concentration may be harmful for human health. Therefore, it is impending to reduce the concentration of metal ions in contaminated lakes and rivers for fishery and also for human health.

Microbiota of Vibrio sp. in the hepatopancreas of cultured white pacific shrimp (Litopenaeus vannamei) / Vibrio sp., microbiota en el hepatopáncreas del camarón blanco del Pacífico (Litopenaeus vannamei)

Objective. The present study aimed to investigate the presence of vibrios in the hepatopancreas of cultured shrimp. Materials and methods. Vibrios from the hepatopancreas of fifteen samples of five specimens each, of apparently healthy Pacific white shrimp (Litopenaeus vannamei) were isolated, identified and quantified. Results. The vibrio density ranged from 430 to 2,400 MPN g-1 (rs MPN cm-1=-0.114; rs MPN g-1 = 0.211). Thirty isolations were obtained, most of which belonged to the species V. cholerae (n=11) and V. parahaemolyticus (n=7). Conclusions. The outcomes of the present study suggest that, even in the absence of symptoms of vibriosis, the microbiota of the hepatopancreas of cultured shrimp may include sucrose positive and negative vibrios.

Objetivo. El presente estudio tuvo como objetivo investigar la presencia de vibrios en el hepatopáncreas del camarón de cultivo. Material y métodos. En este estudio, fueron aislados, identificados y cuantificados los vibrios del hepatopáncreas de 75 camarones blancos del Pacífico (Litopenaeus vannamei), aparentemente sanos, oriundos de un cultivo en la región nordeste de Brasil. Quince muestras, cada una consta de cinco camarones, se pusieron a prueba. Resultados. La densidad de Vibrio varió de 430 a 2.400 NMP g-1 (rs NMP cm-1 = -0.114; rs NMP g-1 = 0.211). Treinta aislamientos fueron obtenidos, la mayoría de los cuales pertenecían a la especie V. cholerae (n=11) y V. parahaemolyticus (n=7). Conclusiones. Los hallazgos del presente estudio sugieren que, incluso en ausencia de síntomas de la vibriosis, la microbiota de lo hepatopáncreas del camarón de cultivo puede incluir vibrios sacarosa positivos y negativos.

Isolation and characterization of infectious Vibrio sinaloensis strainsfrom the Pacific shrimp Litopenaeus vannamei (Decapoda: Penaeidae)

Infectious diseases especially those caused by bacterial and viral pathogens are serious loss factors in shrimp farming. In this study, bacteria were isolated from the gut and hepatopancreas of stressed shrimps obtained from a commercial farm. The isolates were screened on Thiosulfate citrate bile salt sucrose (TCBS) agar plates for the selection of Vibrio species. Presumptive vibrios were characterized through tests for hemolytic and enzymatic activity, hydrophobicity, growth and molecular identification. Three experimental infections were conducted in order to confirm the pathogenicity of selected bacterial strains VHPC18, VHPC23, VHPC24 and VIC30. In the third experimental challenge the LD50 was obtained, it lasted 10 days with 10 shrimp, weighing 6.9±1.1g, per tank. The treatments in triplicate were: (1) saline solution (control group); (2) 2×10(5)CFU/shrimp; (3) 4×10(5)CFU/shrimp; (4) 2×10(6)CFU/shrimp; (5) 4×10(6)CFU/shrimp, and (6) 8×10(6)CFU/shrimp. In all challenges, water parameters measured during the experimental period remained within optimum ranges. Pathogenicity tests confirmed that the mixture of four vibrio isolates, identified as Vibrio sinaloensis, was virulent for L. vannamei. The LD50 value was 1.178×10(5)CFU/g body weight. V. sinaloensis may act as opportunistic pathogens for cultured L. vannamei. Rev. Biol. Trop. 60 (2): 567-576. Epub 2012 June 01.

Las enfermedades de etiología infecciosa, especialmente las causadas por patógenos bacterianos y virales ocasionan graves pérdidas en el cultivo de camarón blanco Litopenaeus vannamei. En este estudio se caracterizo: la actividad enzimática y hemolítica; hidrofobicidad; crecimiento e identificación molecular de vibrios aislados del intestino y hepatopancreas de camarones estresados, obtenidos de una granja comercial, en medio Agar Tiosulfato Citrato Bilis Sacarosa. Además, se realizaron tres infecciones experimentales para confirmar la patogenicidad de las cepas bacterianas seleccionadas VHPC18, VHPC23, VHPC24 y VIC30. En la tercera infección experimental se obtuvo la LD50, el reto duro 10 días, con 10 camarones por tanque con un peso de 6.9±1.1g. Los tratamientos se realizaron por triplicado: (1) solución salina (grupo control); (2) 2×10(5)UFC/camarón; (3) 4×10(5)UFC/camarón; (4) 2×10(6)UFC/camarón; (5) 4×10(6)UFC/camarón y (6) 8×10(6)UFC/camarón. En todos los retos, los parámetros del agua permanecieron dentro de los intervalos óptimos. Las pruebas de patogenicidad confirmaron que la mezcla de cuatro aislados de Vibrio, identificados como Vibrio sinaloensis, fue virulenta para L. vannamei. El valor de la LD50 fue de 1.178×10(5)UFC/g de peso corporal. Los resultados permiten establecer que las cepas de V. sinaloensis pueden actuar como patógenos oportunistas en L. vannamei cultivado.

Vitellogenin levels in hemolymph, ovary and hepatopancreas of the freshwater crayfish Cherax quadricarinatus (Decapoda: Parastacidae) during the reproductive cycle

The freshwater crayfish Cherax quadricarinatus is a tropical species of great interest for aquaculture. Vitellogenin (Vg), a lipoprotein precursor of the vitellum accumulated in spawned eggs, can be synthesized in the ovary and/or hepatopancreas of most crustaceans, being the hemolymph the way for transporting Vg throughout the reproductive cycle. Concentration of Vg in hemolymph, ovary and hepatopancreas of Cherax quadricarinatus adult females was measured by means of ELISA, specifically developed after purifying the native Vg. Measurements were made at four periods of the reproductive cycle: pre-reproductive, mid-reproductive, late reproductive and post-reproductive. Besides, both hepatosomatic (HSI) and gonadosomatic (GSI) indexes were determined in each period. Significant variations in Vg levels were detected in both hemolymph and hepatopancreas, being the highest values observed during the mid-reproductive period. Besides, such variations were positively correlated to the HSI. A positive correlation between Vg levels in hepatopancreas and ovary was also seen. These results support previous evidences about the central role of the hepatopancreas as a site of Vg synthesis in the studied species, together with the relevancy of hemolymph for transporting Vg from the hepatopancreas to the ovary. For aquaculture purposes, Vg monitoring in hemolymph could be used as a non-injurious method, to check the reproductive activity of C. quadricarinatus females.

La langosta de agua dulce Cherax quadricarinatus es una especie tropical de gran interés para la acuicultura. Se midió la concentración de vitelogenina (Vg) en hemolinfa, ovario y hepatopáncreas de hembras adultas de esta especie, por medio de ELISA. Las mediciones fueron hechas en los cuatro períodos del ciclo reproductivo: pre-reproductivo, reproductivo medio, reproductivo tardío y post-reproductivo. Se detectaron variaciones significativas en los niveles de Vg tanto en hemolinfa como en hepatopáncreas, se observó el mayor valor durante el período reproductivo medio. Además, tales variaciones se correlacionaron positivamente con el índice hepatosomático. Se observó además una correlación positiva de los niveles de Vg entre hepatopáncreas y ovario. Estos resultados apoyan evidencias previas sobre el papel central del hepatopáncreas como sitio de síntesis de Vg, en esta especie, y también enfatizan la importancia de la hemolinfa para el transporte de la Vg del hepatopáncreas al ovario. Para propósitos de acuicultura, la medición de Vg en hemolinfa podría ser utilizada como un método no lesivo, con el fin de constatar la actividad reproductiva de hembras de C. quadricarinatus.

Histoarchitectural features of the hepatopancreas of the Amazon River prawn Macrobrachium amazonicum / Histoarquitectura del hepatopáncreas del camarón de la Amazonia Macrobrachium amazonicum

In decapod crustaceans, the digestive gland is concerned with the digestion, absorption of nutrients, the storage of reserves and excretion. The metabolism and the histological and histochemical changes of the hepatopancreas are observed in response to physiological demands as moult, reproduction, digestive process. Thus the hepatopancreas structure should be recognized to provide important morphological information to future studies involving the nutrition requirements of freshwater prawn culture. In this study, second-generation Macrobrachium amazonicum produced from wild broodstock collected in the state of Para in Brazil were used. Thirty adult male and female M. amazonicum were selected and randomly transferred to five experimental units for macroscopic and microscopic studies. The hepatopancreas of M. amazonicum is a large, yellowish-brown, compact organ, which occupies much of the cephalothoracic cavity. It has right and left halves that are enclosed together in a laminar connective tissue capsule, and at the same time they are separated by an interstitial connective tissue. The two halves are thereby called the right and left hepatopancreatic lobes. The principal tubule gives rise to four secondary tubules at each hepatopancreatic lobe. The morphological and functional unit consists of a blind-ended hepatopancreatic tubule, considered in the present study as the hepatopancreatic lobule. Each hepatopancreatic tubule can be subdivided into distal, medial and proximal zones. The hepatopancreatic tubule is lined by a pseudostratified epithelium that consists of five different cell types, which include the E-cell (embryonic), F-cell (fibrillar), B-cell (blister-like), R-cell (resorptive) and M-cell (midgut or basal). It is important to emphasize that the function of each cell type in the hepatopancreas during the digestive cycle is not yet established for decapods.

La glándula digestiva en los crustáceos decápodos asume las funciones de digestión, absorción de nutrientes, almacenamiento de las reservas energéticas y excreción de metabolitos. El metabolismo y las alteraciones histológicas e histoquímicas son observados como respuesta a necesidades fisiológicas, tales como: muda, reproducción y procesos digestivos. Por lo tanto, se requiere conocer la estructura histológica del hepatopancreas con el fin de reunir información morfológica para futuros estudios que consideren las necesidades nutricionales para el cultivo de los camarones de agua dulce. Fueran utilizados 30 animales, machos y hembras de Macrobrachium amazonicum, producidos a partir de reproductores colectados en el Estado de Para, Brasil. Las observaciones macro y microscópicas permitieron concluir que el hepatopancreas del M. amazonicum es un órgano grande y compacto de color amarillo tendiendo a marrón, que ocupa la mayor parte de la cavidad céfalo-torácica. El órgano presenta dos mitades, derecha y izquierda, las cuales están envueltas por una cápsula de tejido conjuntivo, y al mismo tiempo, separadas por tejido conectivo intersticial. Las dos mitades del órgano son identificadas como lobos derecho e izquierdo. Cada lobo presenta un túbulo principal que origina cuatro túbulos secundarios. Por lo tanto, la unidad morfofuncional del hepatopancreas consiste en un túbulo hepatopancreático de fondo ciego, considerado en esta investigación como lóbulo hepatopancreático. Cada lóbulo hepatopancreático puede ser dividido en 3 regiones: distal, media y proximal, y en su totalidad, se observa revestido por epitelio seudo estratificado que reúne cinco tipos celulares: célula E (embrionaria), célula F (fibrilar), célula B (globosa), célula R (reabsortiva) y célula M (basal). Es importante enfatizar que la función de cada tipo celular del hepatopancreas no está todavía aclarada para los decápodos.

Bioactivity of crude ethanol extract and fractions of Eugenia uniflora (Myrtaceae) in the hepatopancreas of Oreochromis niloticus L

This study evaluates the bioactivity of the crude ethanol extract and ethyl acetate, hexane and chloroform fractions obtained from Eugenia uniflora leaves using the hepatopancreas of Oreochromis niloticus L. as an experimental model. The ethanol extract and fractions were administered to the fish orally with their feed. Twenty-four hours later, the fish were sacrificed and their livers dissected, fixed in neutral formalin, embedded in paraffin and sectioned. Histological analyses were performed using Masson's trichrome and Haematoxylin-Eosin. Histochemical studies were performed using Feulgen, PAS (Periodic Acid Schiff) and PAS + salivary amylase and Sudan IV stain. The qualitative analysis of the material showed that the crude extract and the ethyl, chloroform and hexane fractions induced vasodilation, vascular congestion and toxicity due to the presence of eosinophilic granular cells, rodlet cells, some leukocytic infiltrate and rare focal necroses. The Nile tilapia proved to be a satisfactory model for screening plant products.

The effect of crude ethanol extract and fractions of Hyptidendron canum (Pohl ex Benth) Harley on the hepatopancreas of Oreochromis niloticus L

Hyptidendron canum (Pohl ex Benth.) Harley is a native tree of the Brazilian Savannah. The fish Oreochromis niloticus L. was used as an experimental model to determine the bioactivity of the crude ethanol extract as well as ethyl acetate, hexanic and chloroform fractions obtained from its leaves. The plant ethanol extract and fractions were administered to the fish orally with their feed. Twenty four hours later, the fish were sacrificed and their livers dissected, fixed in neutral formalin, embedded in paraffin and sectioned. Histological analyses were performed using Masson's trichrome and Haematoxylin-Eosin. Histochemical studies were performed using Feulgen, PAS (Periodic Acid Schiff) and PAS + salivary amylase and Sudan IV stain. The qualitative analysis of the material showed that both the crude ethanol extract and the fractions from H. canum induced vasoactive activity, causing vasodilation and vascular congestion, and the hexanic fraction also caused an apparent proliferation of capillaries. Hepatopancreas toxicity was evident through inflammatory processes. Pancreatic (chloroform fraction) and hepatic alterations, hemorrhagic spots and necroses were observed in fish treated with-ethanol extract and fractions. This study is the first description of the biologic action of the crude ethanol extract and the hexane, ethyl acetate and chloroform fractions in fish.

Neuroendocrine regulation and pesticidal impact on freshwater crab, Barytelphusa guerini (H. Milne Edwards).

The extensive use of pesticides to control agricultural pests poses a serious threat to many non-target organisms of the aquatic environment such as the freshwater crab, B. guerini. The deleterious influence of the pesticide causes physiological, biochemical, histological and such other disorders in the animal exposed. In the present study impact of an organophosphate pesticide, monocrotophos and neuroendocrine regulation on the biochemical contents of hepatopancreas of B. guerini has been studied. Experimental studies revealed that glycogen and protein content decreased in normal crabs when exposed to sub lethal concentrations of monocrotophos, while lipid content was increased. In the ablated and pesticide exposed crabs glycogen, protein and lipid contents decreased. In case of ablated and exposed crabs when injected with eyestalk extract, glycogen and protein contents declined, whereas lipid content hiked. It was observed that glycogen, protein and lipid contents in eyestalk extract injected crabs were similar to those of normal exposed crabs. This indicates the vital role of eyestalk in the regulation of biochemical contents. Histological studies of the hepatopancreas indicate structural changes such as large number of vacuolated cells and phagocytes when exposed to the pesticide.

Heavy metal accumulation in certain marine animals along the East Coast of Chennai, Tamil Nadu, India.

Heavy metals disposed through anthropogenic activities find their way into the oceans and seas through the rivers or through direct fall out from factory effluents. These heavy metals resuspend back into the water column along with the sediments and are known to affect the marine animals. Marine animals like fish, prawn, crab and mussel were collected along the East Coast (off Pulicat lake to Chennai Harbour) to evaluate trace metal concentrations in various tissues. The above specimens accumulated heavy metals such as Zn, Pb, Cu, Co, Cr, Ni and Cd. Fish, prawn, crab and mussel revealed higher concentration of heavy metals such as Zn, Pb, Cr Co, Cu and Ni and Cd in low levels. The results revealed that the heavy metal concentrations in the marine animals are below the threshold levels associated with the toxicological effects and the regulatory limits. The bioconcentration factors revealed that the animals have accumulated heavy metals along the food chain rather than from the water column and sediment.

Effect of Pseudomonas aeruginosa on the giant freshwater prawn, Macrobrachium rosenbergii--histopathological and electron microscopic study.

The giant freshwater prawn Macrobrachium rosenbergii was injected with an inoculum containing LD, 96 hr dose of 10' Pseudomonas aeruginosa (MTCC 1688) to determine the histopathological effects in vivo. The comparison of tissues of both the control and the bacterial endotoxin treated prawns after 96 hr revealed significant degenerative changes in treated prawns. Both light microscopic and electron microscopic observations revealed the infiltration of the tissues of Pseudomonas sp in the muscular and hepatopancreatic tissues of prawn. The muscular tissue changes in the myofibrillar arrangement with blockage at the gap junctions and necrotic lesions were observed. The hepatopancreatic cells were vacuolated with hypertrophied nucleus. Atrophy of hepatopancreatic tubules was conspicuous. The pathogenicity of Pseudomonas aeruginosa is attributed to its infiltration and multiplication inside the tissues and the consequent release of extra-cellular enzymes for its metabolism. The degeneration of host tissues is also attributed to the latter.

Functional morphology of the hepatopancreas of Palaemonetes argentinus (Crustacea: Decapoda): influence of environmental pollution

We analyzed the morphological and functional state of hepatopancreas in Palaemonetes argentinusfrom two environments with different pesticide concentrations. Los Padres lagoon (Argentina) is an area subjectedto contamination due to the slow exchange of water, the shallow depth and the input of contaminatedwater. Prawns living in this lagoon accumulate high amounts of organochlorine pesticides in their tissues. Hepatopancreas of prawns from Canal 5, an adjacent shallow stream where the amount of pesticides is belowtoxic levels, and from Los Padres lagoon were processed by standard histological techniques with light microscopyand transmission electronic microscopy. At Los Padres lagoon, we found important tissular alterations,such as intertubular infiltration of haemocytes and connective tissue, epithelial retraction in some tubules, anda folded basal lamina. Important necrotic desquamation, with cariolysis, cariorrexis and lack of cellular detailswere also observed. Numerous tubules presented an enlarged and irregular lumen with the epithelium atrophiedor completely absent. In general, the lesions were particularly located in the medullar region of the organ. Atthe ultrastructural level, R and F cells were the most damaged. Both cell types had nuclear retraction, chromatincondensation and cytoplasmic lysis. Some R cells also had dilated mitochondria and numerous lysosomes, and the basal cytoplasm was nearly completely lysed. The hepatopancreas of prawns from Canal 5 did not evidenceany alterations. The histopathological study of the hepatopancreas is a highly sensitive tool to evaluate thephysiological condition of prawns and water quality. Other environmental conditions were similar, so it can beassumed that pollutants were the main cause of organ deterioration.

Analizamos el estado morfológico y funcional del hepatopáncreas de Palaemonetes argentinus de dosambientes con diferentes concentraciones de plaguicidas. La laguna Los Padres (Argentina) es un área sujeta a contaminación debido al lento intercambio del agua, la escasa profundidad y el influjo de agua contaminada. Las gambas acumulan aquí grandes cantidades de plaguicidasorganoclorados en sus tejidos. Los hepatopáncreas de gambas del Canal 5 y de la laguna Los Padres fueronprocesados mediante técnicas histológicas estándar para microscopía óptica y electrónica de transmisión. Loshepatopáncreas de los individuos recolectados en Los Padres tenían alteraciones tisulares importantes, comoinfiltración intertubular de hemocitos y tejido conectivo, retracción epitelial en algunos túbulos y láminas basalesplegadas. También se observó descamación necrótica importante, con cariolisis, cariorrexis y falta de detallescelulares. Vimos muchos túbulos con un lúmen irregular y agrandado, con el epitelio atrofiado o completamenteausente. En general las lesiones se localizaron en la región medular del órgano. Ambos tipos celulares mostraronretracción del núcleo, condensación de la cromatina y ruptura del citoplasma. Algunas células R también teníanmitocondrias y numerosos lisosomas dilatados, y el citoplasma basal casi completamente desintegrado. Nohallamos alteraciones en los hepatopáncreas de gambas deCanal 5. El estudio histopatológico del hepatopáncreas es una herramienta muy apropiada para evaluar la condiciónfisiológica de las gambas y la calidad del agua. Por la semejanza de otras condiciones ambientales, suponemosque los contaminantes fueron la causa principal del deterioro de los órganos.

Fine structural analysis of the epithelial cells in the hepatopancreas of Palaemonetes argentinus (Crustacea, Decapoda, Caridea) in intermoult

The aim of this study is to describe the ultrastructure of the hepatopancreas of P argentinus in intermoult. P argentinus hepatopancreas was studied using standard TEM techniques. Each tubule consists of four cellular types: E (embryonic), F (fibrillar), R (resorptive) and B (blister like). E-cells have embryonic features and some of them were found in mitosis. F, R and B cells possess an apical brush border. F-cells have a central or basal nucleus, a conspicuous RER, and dilated Golgi cisternae. R cells show a polar organization of organelles in three areas: apical, with numerous mitochondria and sER tubules, a central are a with the nucleus and RER, and a basal area containing a sER-like tubule system and mitochondria. B-cells were observed at different stages of their life cycle. In an early differentiation stage they comprise an apical endocytotic complex and Golgi vesicles. The fusion of endocytotic and Golgi vesicles originates subapical vacuoles. During maturation, a big central vacuole is formed by coalescence of subapical vacuoles. The central vacuole is eliminated by holocrine secretion. The ultrastructure suggests that F-cells synthesize proteins, R-cells storage nutrients and B-cells have a secretory or excretory function, and confirms the independent origin of F, B and R cells from the embryonic cells

Effect of Pseudomonas aeruginosa (MTCC 1688) on the tissue phosphatases activity on Macrobrachium rosenbergii (De Man).

A time course study on the endotoxin toxicity of the gram negative bacteria, Pseudomonas aeruginosa MTCC 1688 on the tissue phosphatases activity on the giant freshwater prawn, Macrobrachium rosenbergii was conducted. The results revealed marked elevation of both acid and alkaline phosphatase activity in the haemolymph and body muscle. The hepatopancreas showed reduced phosphatase activity compared to control. The enzymes, being non-specific in action and particularly the acid phosphatase being of lysosomal origin, their increase in muscle and haemolymph has pathogenic significance in the inoculum treated prawns.