mast cells in the intestine and gills of the sea bream, sparus aurata, exposed to a polychlorinated...

G

A

Mp

ESa

b

c

a

ARRAA

KMPGISST

I

tratmi

pmiaat

fP

0d

ARTICLE IN PRESSModel

CTHIS-50501; No. of Pages 6

Acta Histochemica xxx (2011) xxx–xxx

Contents lists available at ScienceDirect

Acta Histochemica

journa l homepage: www.e lsev ier .de /ac th is

ast cells in the intestine and gills of the sea bream, Sparus aurata, exposed to aolychlorinated biphenyl, PCB 126

ugenia Rita Laurianoa, Margherita Calòb, Giuseppa Silvestrib, Daniele Zacconec,∗,imona Pergolizzi a, Patrizia Lo Cascioa

Department of Food and Environmental Science, Faculty of Science FF.MM.NN., University of Messina, I-98166 Messina, ItalyDepartment of Experimental Sciences and Applied Biotechnology, Faculty of Veterinary Medicine, University of Messina, I-98168 Messina, ItalyDepartment of Animal Biology and Marine Ecology, Faculty of Science, University of Messina, I-98166 Messina, Italy

r t i c l e i n f o

rticle history:eceived 9 December 2010eceived in revised form 12 April 2011ccepted 13 April 2011vailable online xxx

eywords:ast cells

CB 126

a b s t r a c t

The presence of mast cells has been reported in all classes of vertebrates, including many teleost fishfamilies. The mast cells of teleosts, both morphologically and functionally, show a close similarity to themast cells of mammals. Mast cells of teleosts, localized in the vicinity of blood vessels of the intestine,gills and skin, may play an important role in the mechanisms of inflammatory response, because theyexpress a number of functional proteins, including piscidins, which are antimicrobical peptides that actagainst a broad-spectrum of pathogens. An increase in the number of mast cells in various tissues andorgans of teleosts seems to be linked to a wide range of stressful conditions, such as exposure to heavymetals (cadmium, copper, lead and mercury), exposure to herbicides and parasitic infections. This study

illsntestineea breamparus aurataeleosts

analyzed the morphological localization and abundance of mast cells in the intestine and gills of seabream, Sparus aurata, after a 12, 24 or 72 h exposure to PCB 126, a polychlorinated biphenyl, which is apotent immunotoxic agent. In the organs of fish exposed to PCB 126, it was observed that in addition tocongestion of blood vessels, there was extravasation of red blood cells, infiltration of lymphocytes, anda progressive increase in numbers of mast cells. These data confirm the immunotoxic action of PCB, and

cells
the involvement of mast
ntroduction

All vertebrates, including teleost fish, are subject to stress andheir immune systems are affected by dramatic changes in the envi-onment including pollutants. Polychlorinated biphenyls (PCBs)re persistent environmental pollutants that have been reportedo induce proliferation of mast cells (MCs), rodlet cells (RC) and

ucous cells (Dezfuli et al., 2003, 2010; Quabius et al., 2005) in fishmmune systems.

Polychlorinated biphenyls (PCBs) are a class of organic com-ounds with 1–10 chlorine atoms attached to biphenyl, which is aolecule composed of two benzene rings. PCBs were widely used

n the synthesis of pesticides, herbicides, solvents, disinfectants

Please cite this article in press as: Lauriano ER, et al. Mast cells in the intestinebiphenyl, PCB 126. Acta Histochemica (2011), doi:10.1016/j.acthis.2011.04

nd especially as dielectric fluids in transformers and capacitors,nd are still used for some industrial processes. Two major struc-ural classes of PCBs include the co-planar PCBs, as 3, 3′, 4, 4′,

Abbreviations: EGCs, eosinophilic granular cells; HP, hyperplasia; LF, lamellarusion; MCs, mast cells; MGG, May-Grünwald-Giemsa; MT, Masson’s trichrome;CBs, polychlorinated biphenyls; RC, rodlet cells; SW, swelling.∗ Corresponding author.

E-mail address: [email protected] (D. Zaccone).

065-1281/$ – see front matter © 2011 Elsevier GmbH. All rights reserved.oi:10.1016/j.acthis.2011.04.004

in the inflammatory response.© 2011 Elsevier GmbH. All rights reserved.

5-pentachlorinated biphenyl (PCB 126) (Hestermann et al., 2000)and non-coplanar as PCB 153. PCB 126 is considered one of themost potent dioxin-like and potent immunotoxic agents (Calò et al.,2010). The dioxins are organochlorine persistent, toxic and bioac-cumulative chemicals. The health effects of PCBs have been widelystudied in humans, laboratory animals, and in wildlife in con-taminated areas. High-level exposure to selected organochlorinesappears to cause abnormalities of liver function, skin (chloracne),nervous system, endocrine system, reproductive system, immunesystem, and may cause cancer (Longnecker et al., 1997). Cellularcomponents of innate immunity (rodlet cells) are found in the bul-bus of the piscine heart (Reite, 1997), however, the effects of PCBson fish cardiovascular systems are unknown.

Mast cells (MC), also known as eosinophilic granular cells(EGCs), are present in most species of teleosts and are found ina variety of tissues, including the gut, gills, skin, brain, and in thevicinity of blood vessels (Reite, 1997; Murray et al., 2007). Fish mastcells are analogous to mammalian mast cells in their cytochemicalcharacteristics and their function (Dezfuli and Giari, 2008), how-

and gills of the sea bream, Sparus aurata, exposed to a polychlorinated.004

ever, the homology of EGCs in fish to mammalian MCs remains acontroversial issue.

The general morphology, staining characteristics and tissuedistribution together with discussion of some of the proposed

dx.doi.org/10.1016/j.acthis.2011.04.004


http://www.sciencedirect.com/science/journal/00651281

http://www.elsevier.de/acthis

mailto:[email protected]


ING Model

A

2 istoch

f2impMtciE1e

lr4ttaP2dt2tbesel(e

M

E

wwac

esfl

sdetw1ipc(sPr

H

p

ARTICLECTHIS-50501; No. of Pages 6

E.R. Lauriano et al. / Acta H

unctions of these cells have been reviewed (Reite and Evensen,006). EGCs are considered to be a component of the teleost innate

mmune system and have been observed in regions of active inflam-atory responses, especially those associated with bacterial and

arasitic infections (Reimschuessel et al., 1987; Sharp et al., 1989;atsuyama and Iida, 1999; Dezfuli et al., 2003). These cells tend

o accumulate in large numbers in tissues exposed to pathologicalhallenges and have been observed to exhibit distinct morpholog-cal changes, including near complete degranulation (Vallejo andllis, 1989; Lamas et al., 1991; Powell et al., 1991, 1993; Flano et al.,996; Noya and Lamas, 1997; Holland and Rowley, 1998; Schmalet al., 2004).

The presence of some morphological and histochemical simi-arities to mammalian MCs (Reite, 1997), including the observedesponse to known mast cell degranulating agents (i.e. compound8/80) and effects on vascular permeability, have led researcherso propose a role for these cells in the induction of inflamma-ory responses. These include vasodilation, neutrophil attractionnd migration and macrophage activation (Vallejo and Ellis, 1989;owell et al., 1991, 1993; Reite, 1997; Matsuyama and Iida, 1999,001). Studies have also shown that EGCs are involved in the pro-uction of specific antimicrobial peptides and are therefore thoughto be directly involved in killing microbes (Silphaduang and Noga,001; Iijima et al., 2003; Murray et al., 2003). Teleost gills and intes-ine are considered important immune organs and therefore shoulde able to mount an immune reaction (Paulsen et al., 2001; Penissit al., 2003; Smith and Fernandes, 2009). These organs are also con-idered as important portals of entry for fish pathogens (Mulerot al., 2008; Ellis, 2001). In this study we analyzed the morpho-ogical localization and local increase in the number of mast cellsMCs) in the intestine and gills of the sea bream, Sparus aurata, afterxposure to PCB126a for 12, 24 or 72 h.

aterials and methods

xperimental protocol

Twenty juvenile sea breams (S. aurata), weighing about 30 g,ere used in the study. All animal procedures were in agreementith the Declaration of Helsinki and with the Guide for the Care

nd Use of Laboratory Animals. Protocols for all experimental pro-edures were approved by the local Ethics Committee.

Fish, were obtained from the “Acquacoltura Lampedusa” hatch-ry, Messina, Italy. Before experimentation, the fishes were kept inea water tanks at 12–15 ◦C provided by the Experimental Centeror Ichthyology (CISS), University of Messina, and acclimatized toaboratory conditions for 10 days. Sea breams were not fed.

Fishes were equally divided into two groups and allocated ineparate bags. The first group received PCB 126 (10−8 M dissolved inimethyl sulfoxide-DMSO) (Lookchem, Hangzhou, China) (Jönssont al., 2003); and the second group, used as controls, received justhe vehicle used to dissolve the PCBs (DMSO). For PCB exposuree used transparent polyethylene bags (50 cm × 70 cm) filled with

5 liters of continuously aerated sea water. The bags were placedn plastic boxes (45 cm × 30 cm × 15 cm). During the experiment,H and oxygen dissolved in sea water were monitored. During theourse of experiment, five animals of each group were anesthetizedMS-222, tricaine methanesulfonate, Sigma–Aldrich, 200 mg/L ofea water) and sacrificed after 12 h and at 24 h of exposure toCBs and after 72 h following removal of PCBs. Gills and gut wereemoved for histological analysis and kept in physiological saline.


istology

For the histological examinations, the samples were fixed in 4%araformaldehyde in phosphate buffered saline (PBS) 0.1 M (pH

PRESSemica xxx (2011) xxx–xxx

7.4) for 2–4 h, dehydrated in graded ethanol, cleared in xylene,embedded in Paraplast® (McCormick Scientific LLC, St. Louis, MO,USA) and cut into 5 �m sections. The sections were stained withhematoxylin–eosin (H & E), Masson’s trichrome, alcian blue (pH2.5)-PAS (periodic acid-Schiff), toluidine blue (pH 0.5 and pH 1.0)and May-Grünwald-Giemsa (MGG). Sections were examined undera Zeiss Axioskop 2 plus microscope equipped with a Sony DigitalCamera DSC-85.

Results

The fish gills of the species studied consist of gill arches, gill fila-ments (primary lamellae) and gill lamellae (secondary lamellae).The gill arches and filaments are supported by a branchial sys-tem of cartilaginous rods. Different cell types are associated withthe gill epithelium and submucosal layers namely: goblet cells,chloride cells, macrophages, eosinophilic granular cells and lym-phocytes. Some of these cell types are shared in common with someimmune-related associated cells in the intestinal epithelium. Theidentification of MCs/EGC in the gills and intestine is in agreementwith that reported by Dickerson (2009). In the gills and intestineof fish exposed to PCB 126 there was a progressive increase innumber of mast cells, which were full of numerous cytoplasmicgranules strongly stained with eosin (Figs. 1b and 2b). The granulesof MCs/EGCs stained with MGG were eosinophilic as previouslyreported by Reite and Evensen (2006) (Figs. 1d, 2g and h). In thehistological sections of gills and intestine stained with Masson’strichrome, MC granules were stained red (Fig. 1c). MCs/EGCs showno metachromatic staining with toluidine blue at lower pH values,which is in agreement with Mulero et al. (2007). The classifica-tion of these cells was also made using their histochemical feature(a faint to appreciable alcianophilia). The sections of the gills andintestine from control fish revealed the presence of a few mast cells(Figs. 1a and 2a). There was a moderate increase of mast cells in theconnective tissue surrounding the central cartilage of the gill fila-ments, and in the submucosal layer of the intestine (Figs. 1b and 2b)in fish exposed to PCB 126 at 12 h. At 24 and 72 h, PCB 126 causeda persistent local inflammatory reaction and a large number ofMCs/EGCs were present (Figs. 1c, d, 2c and d). Degranulation ofmast cells was frequent, and characterized by conspicuous swellingof granules and often by the presence of free granules adjacent tothe mast cells (Fig. 2c).

Discussion

The mucosal surfaces of the skin, gills and intestine of fishesare constantly exposed to environmental pathogens. Resistanceto infection and recovery is facilitated in part by innate non-specific immunity that consists of a plethora of constitutivelyexpressed elements as well induced components of the inflam-matory response (for review see: Dickerson, 2009). The MCs/ECGscomprise a part of the innate cell inflammatory response in teleosts.The gastrointestinal-associated lymphoid tissue (GALT) of teleostsmainly consists of lymphocytes, plasma cells, macrophages aswell as different types of granulocytes, both acidophilic and neu-trophilic (Du Pasquier and Litman, 2000; Mulero et al., 2007). EGCsare present and may serve to modulate immune-hypersensitiveresponses that occur in the gut (Dickerson, 2009). MCs/EGCs werelocated in the lamina propria in the gut of various teleost species(Rombout et al., 1989; Leckness, 2002). In salmonids MCs/EGCs areprincipally located in the submucosal layers of the intestine and


move into the villi or mucosa in certain bacterial infections asso-ciated by a degranulation of these cells (Ellis et al., 1981). Thesecells may play an important role in innate immunity and immunesurveillance and studies have shown that the administration of


ARTICLE IN PRESSG Model

ACTHIS-50501; No. of Pages 6

E.R. Lauriano et al. / Acta Histochemica xxx (2011) xxx–xxx 3

Fig. 1. Progressive increase of mast cells in gills of fish exposed to PCB 126 (10−8 M). (a) The gills of control fish stained with H & E show no histopathological changes. A fewmast cells (a resident cell population) are associated with the loose connective of the gill arch. (b) In fish exposed for 12 h, moderate increase of mast cells (MC) in connectiveof the gill arch and in the primary lamellae H/E. (c) Masson’s trichrome stain in the gill exposed for 24 h reveals lamellar fusion (LF) and a strong increase of MCs with granuless MGG)e figure

ptntcftEtBaskhcneawe

tained red. (d) Gills of fish exposed for 72 h, stained with May-Grünwald-Giemsa (pithelium (scale bar = 10 �m). (For interpretation of the references to color in this

robiotics to gilthead sea bream elicits an increased number ofhese cells in the gut (Picchietti et al., 2007). Contrary to the intesti-al epithelium, the function of the various types of leukocytes inhe gills has been little investigated. Their roles are presumably toapture foreign substances and kill infectious agents that gain entryrom the water. The gill phagocytes also apparently participate inhe clearance of foreign substances from the blood (Leckness, 2002).xperimental studies showing the involvement of phagocytic cellshat migrate to gill tissues during inflammation are very scarce.ioaccumulation of PCB 126 results in persistent levels in animalnd human tissues and the biological responses to PCB 126 areimilar to those of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), anown human carcinogen. PCB congeners or mixtures (Aroclors)ave been associated with various toxic responses including: can-ers, hepatotoxicity, gastrointestinal and respiratory disturbances,eurotoxicity, immunotoxicity, reproductive and developmental


ffects, and endocrine disruption including thyroid toxicity (Lind Hansen, 1996). In this study, a large number of mast cellsere observed in the gills and intestine of sea breams (S. aurata)

xposed to PCB 126. Recently, two inflammatory cell types found

, showed lamellar fusion (LF), hyperplasia (HP) and eosinophilic material under thelegend, the reader is referred to the web version of the article.)

in teleosts have been studied: mast cells (MCs/ECGs) and rodletcells (RC) (Dezfuli et al., 2003, 2008; Reite and Evensen, 2006;Silphaduang et al., 2006). The origin of mast cells in vivo is notclear. They probably differentiate in hematopoietic organs andreach their target organs by the blood circulation as immaturecells (Bergeron and Woodward, 1983; Powell et al., 1990; Flanoet al., 1996), as has been documented for mammalian mast cells(Kirshenbaum et al., 1991). Mast cells are present in the alimen-tary canal, skin, and gills of the majority of teleosts (Mazon et al.,2007; Ezeasor and Stokoe, 1980; Ellis, 1985; Murray et al., 2007)and in regions of active inflammatory responses, especially dueto bacterial and parasitic infections (Reimschuessel et al., 1987;Sharp et al., 1989; Dezfuli and Giari, 2008; Dezfuli et al., 2008,2009).

Mast cells produce specific antimicrobial peptides, namelypleurocidin (Secombes, 1996; Murray et al., 2007) and piscidin


(Silphaduang and Noga, 2001; Noga and Silphaduang, 2003;Silphaduang et al., 2006). Histamine is also stored in mast cellsof Perciformes, the most evolutionarily advanced order of teleosts(Dezfuli et al., 2010).


ARTICLE IN PRESSG Model

ACTHIS-50501; No. of Pages 6

4 E.R. Lauriano et al. / Acta Histochemica xxx (2011) xxx–xxx

Fig. 2. Progressive increase of mast cells in the intestine of fish exposed to PCB 126 (10−8 M). (a) No histopathological changes in the intestine of control fish stained withMay-Grünwald-Giemsa (MGG). A few mast cells are seen in the connective tissue. (b) In fish exposed for 12 h, moderate increase of mast cells (MC) in the submucosa layerw ed for( nectiv

owwtaeC

lthw(a

oslti

as observed. H & E. (c) May-Grünwald-Giemsa (MGG) in the intestine in fish exposd) The prolonged exposure (72 h) caused a significant increase of mast cells in con

In the present study, no histopathological changes werebserved in the gills of the control fish. A few mast cells associatedith the loose connective of the gill arch and the primary lamellaeere observed, suggesting a resident cell population. This suggests

hat the gills are important immunological organs and producerapid response to any pathogenic or parasitic challenge (Flano

t al., 1996; Noya and Lamas, 1997; Holland and Rowley, 1998;ampos-Perez et al., 2000; Dezfuli et al., 2003).

In the organs exposed to PCB 126 at 24 h and 72 h, histologicalesions were observed in gill lamellae including edema, separa-ion of epithelium from lamellae (epithelial lifting), lamellar fusion,yperplasia of lamella and the space under the epithelium filledith eosinophilic material. Similar findings were also observed

Sunitha and Sahai, 1983; Roy and Munshi, 1991; Saravana Bhavannd Geraldine, 2000).

The prolonged exposure to PCBs caused a significant increasef mast cells in connective tissue and blood vessels of primary and


econdary lamellae of gills, and close to blood vessels of the muscu-aris layers of the intestine. The close association of mast cells withhe capillary endothelial cells and their presence in gill capillar-es suggests that they may migrate across the endothelium (Powell

24 h showed degranulation of MC and conspicuous swelling (SW) of their granules.e tissue of the muscularis layers of the intestine (MGG) (scale bar = 10 �m).

et al., 1990; Reite, 1997; Holland and Rowley, 1998; Murray et al.,2007). According to Dezfuli and Giari (2008), fish may have twopopulations of mast cells, circulating and resident, and the expo-sure to immunotoxic agents induces recruitment of mast cells tothe sites of inflammation.

Many studies have demonstrated how mast cells degranulate inresponse to exposure to a variety of known degranulating agentsand pathogens with release of their contents (Ellis, 1985; Sharpet al., 1989; Vallejo and Ellis, 1989; Powell et al., 1990, 1993;Flano et al., 1996; Reite, 1997; Paulsen et al., 2001; Schmale etal., 2004; Murray et al., 2007; Dezfuli and Giari, 2008; Dickerson,2009). Some researchers have proposed that mast cell secre-tion may play a role in attracting other types of fish cells (e.g.neutrophils in gills of sea bream) involved in the inflammatoryprocess (Powell et al., 1991; Matsuyama and Iida, 1999; Reite andEvensen, 2006; Mulero et al., 2007). In the present study, intensedegranulation of MCs was observed in gills and intestine, such as


mast cells migration from gill filament to lamellae in fishes sub-jected to prolonged exposure. Our results confirm the immunotoxicaction of PCBs, and the involvement of MCs in the inflammatoryresponse.


ING Model

A

istoch

R

B

C

C

D

D

D

D

D

D

D

E

E

E

E

F

H

H

I

J



eferences

ergeron TCM, Woodward W. Ultrastructure of the granule cellsin the small intestine of the rainbow trout (Salmo gairdneri)before and after stratum granulosum formation. Can J Zool1983;61:133–8.

alò M, Alberghina D, Bitto A, Lauriano ER, Lo Cascio P. Estrogenicfollowed by anti-estrogenic effects of PCBs exposure in juvenilefish (Sparus aurata). Food Chem Toxicol 2010;48:2458–63.

ampos-Perez JJ, Ward M, Grabowski PS, Ellis AE, Secombes CJ.The gills are an important site of iNOS expression in rainbowtrout Oncorhynchus mykiss after challenge with the gram-positive pathogen Renibacterium salmoninarum. Immunology2000;99:153–61.

ezfuli BS, Giari L, Simoni E, Palazzi D, Manera M. Alteration ofrodlet cells in chub caused by herbicide Stam® M-4 (Propanil).J Fish Biol 2003;63:232–9.

ezfuli BS, Giari L. Mast cells in the gills and intestines of naturallyinfected fish: evidence of migration and degranulation. J Fish Dis2008;31:845–52.

ezfuli BS, Giovinazzo G, Lui A, Giari L. Inflammatory response toDentitruncus truttae (Acanthocephala) in the intestine of browntrout. Fish Shellfish Immunol 2008;24:726–33.

ezfuli BS, Manera M, Giari L. Immune response to nematode larvaein the liver and pancreas of minnow, Phoxinus phoxinus (L.). J FishDis 2009;32:383–90.

ezfuli BS, Pironi F, Giari L, Noga EJ. Immunocytochemical localiza-tion of piscidin in mast cells of infected seabass gill. Fish ShellfishImmunol 2010;28:476–82.

ickerson HW. The biology of teleost mucosal immunity. In: Zac-cone G, Perrière C, Mathis A, Kapoor BG, editors. Fish defenses,vol. 2: pathogens, parasites and predators. Enfield: Science Pub-lishers; 2009. p. 1–42.

u Pasquier L, Litman GW. Origin and evolution of the vertebrateimmune system, vol. 248. Berlin: Springer-Verlag; 2000. p. 93.

llis AE. Eosinophilic granular cells (EGC) and histamine responsesto Aeromonas salmonicida toxins in rainbow trout. Dev CompImmunol 1985;9:251–60.

llis AE. Innate host defense mechanisms of fish against viruses andbacteria. Dev Comp Immunol 2001;25:827–39.

llis AE, Hastings TS, Munro ALS. The role of Aeromonas salmonicidaextracellular products in the pathology of furunculosis. J Fish Dis1981;4:41–52.

zeasor DN, Stokoe WM. A cytochemical, light and electron micro-scopic study of the eosinophilic granule cells in the gut ofthe rainbow trout, Salmo gairdneri Richardson. J Fish Biol1980;17:619–34.

lano E, López-Fierro P, Razquin E, Villena A. In vitro differentiationof eosinophilic granular cells in Renibacterium salmoninarum-infected gill cultures from rainbow trout. Fish Shellfish Immunol1996;6:173–84.

estermann EV, Stegeman JJ, Hahn ME. Relative contributions ofaffinity and intrinsic efficacy to aryl hydrocarbon receptor ligandpotency. Toxicol Appl Pharmacol 2000;168:160–72.

olland JW, Rowley AF. Studies on the eosinophilic granulecells in the gills of the rainbow trout Oncorhynchus mykiss.Comp Biochem Physiol C: Pharmacol Toxicol Endocrinol1998;120:321–8.

ijima N, Tanimoto N, Emoto Y, Morita Y, Uematsu K, Murakami T,Nakai T. Purification and characterization of three isoforms ofchrysophsin, a novel antimicrobial peptide in the gills of the redsea bream, Chrysophrys major. Eur J Biochem 2003;270:675–86.


önsson M, Abrahamson A, Brunström B, Brandt I, Ingebrigtsen K,Jørgensen EH. EROD activity in gill filaments of anadromousand marine fish as a biomarker of dioxin-like pollutants. CompBiochem Physiol C: Toxicol Pharmacol 2003;136:235–43.

PRESSemica xxx (2011) xxx–xxx 5

Kirshenbaum AS, Kessler SW, Goff JP, Metcalfe DD. Demonstrationof the origin of human mast cells from CD34+ bone marrowprogenitor cell. J Immunol 1991;146:1410–5.

Lamas J, Bruno DW, Santos Y, Anadon R, Ellis AE. Eosinophilicgranular cell response to intraperitoneal injection with Vib-rio anguillarum and its extracellular products in rainbow troutOncorhynchus mykiss. Fish Shellfish Immunol 1991;1:187–94.

Leckness IL. Uptake of foreign ferritin in platy Xiphophorus macula-tus (Poeciliidae: Teleostei). Dis Aquat Organ 2002;51:233–7.

Li MH, Hansen LG. Enzyme induction and acute endocrineeffects in prepubertal female rats receiving environmen-tal PCB/PCDF/PCDD mixtures. Environ Health Perspect1996;104:712–22.

Longnecker MP, Rogan WJ, Lucier G. The human health effects ofDDT (dichlorodiphenyltrichloroethane) and PCBs (polychlori-nated biphenyls) and an overview of organochlorines in publichealth. Annu Rev Public Health 1997;18:211–44.

Matsuyama T, Iida T. Degranulation of eosinophilic granular cellswith possible involvement in neutrophil migration to site ofinflammation in tilapia. Dev Comp Immunol 1999;23:451–7.

Matsuyama T, Iida T. Influence of tilapia mast cell lysate on vascularpermeability. Fish Shellfish Immunol 2001;11:549–56.

Mazon AF, Huising MO, Taverne-Thiele AJ, Bastiaans J, Verburg-van Kemenade BML. The first appearance of rodlet cells incarp (Cyprinus carpio L.) ontogeny and their possible rolesduring stress and parasite infection. Fish Shellfish Immunol2007;22:27–37.

Mulero I, Sepulcre MP, Meseguer J, Garcia-Ayala A, Mulero V. His-tamine is stored in mast cells of most evolutionarily advancedfish and regulates the fish inflammatory response. Proc NatlAcad Sci U S A 2007;104:19434–9.

Mulero I, Noga EJ, Meseguer J, Garcia-Ayala A, Mulero V. Theantimicrobial peptides piscidins are stored in the granules ofprofessional phagocytic granulocytes of fish and are deliveredto the bacteria-containing phagosome upon phagocytosis. DevComp Immunol 2008;32:1531–8.

Murray HM, Gallant JW, Douglas SE. Cellular localization of pleuro-cidin gene expression and synthesis in winter flounder gill usingimmunohistochemistry and in situ hybridization. Cell Tissue Res2003;312:197–202.

Murray HM, Leggiadro CT, Douglas SE. Immunocytochemicallocalization of pleurocidin to the cytoplasmic granules ofeosinophilic granular cells from the winter flounder gill. J FishBiol 2007;70(Suppl. C):336–45.

Noga EJ, Silphaduang U. Piscidins: a novel family of peptide antibi-otics from fish. Drug News Perspect 2003;16:87–92.

Noya M, Lamas J. Response of eosinophilic granule cells of giltheadseabream (Sparus aurata: Teleostei) to bacteria and bacterialproducts. Cell Tissue Res 1997;287:223–30.

Paulsen SM, Sveinbjornsson B, Robertsen B. Selective staining anddisintegration of intestinal eosinophilic granule cells in Atlanticsalmon after intraperitoneal injection of the zinc chelator dithi-zone. J Fish Biol 2001;58:768–75.

Penissi AB, Rudolph MI, Piezzi RS. Role of mast cells in gastroin-testinal mucosal defense. Biocell 2003;27:163–72.

Picchietti S, Mazzini M, Taddei AR, Renna R, Fausto AM, MuleroV, et al. Effects of administration of probiotic strains on GALTof larval gilthead seabream: immunohistochemical and ultra-structural studies. Fish Shellfish Immunol 2007;22:57–67.

Powell MD, Wright GM, Burka JF. Eosinophilic granule cells in thegills of rainbow trout Oncorhynchus mykiss: evidence of migra-tion? J Fish Biol 1990;37:495–7.


Powell MD, Wright GM, Burka JF. Degranulation of eosinophilicgranule cells induced by capsaicin and substance P in the intes-tine of the rainbow trout Oncorhynchus mykiss (Walbaum). CellTissue Res 1991;226:469–74.


ING Model

A

6 istoch

P

Q

R

R

R

R

R

S



owell MD, Briand HA, Wright GM, Burka JF. Rainbow trout(Oncorhynchus mykiss Walbaum) intestinal eosinophilic gran-ule cells (EGC) response to Aeromonas salmonicida and Vibrioanguillarum extracellular products. Fish Shellfish Immunol1993;3:279–89.

uabius ES, Krupp G, Secombes CJ. Polychlorinated biphenyl 126affects expression of genes involved in stress–immune interac-tion in primary cultures of rainbow trout anterior kidney cells.Environ Toxicol Chem 2005;24:3053–60.

eimschuessel R, Bennett RO, May EB, Lipsky MM. Eosinophilicgranular cell response to a microsporidian infection in a sergeantmajor fish Abudefduf saxatilis L. J Fish Dis 1987;10:319–22.

eite OB. Mast cells/eosinophilic granule cells of salmonids: stain-ing properties and responses to noxious agents. Fish ShellfishImmunol 1997;7:567–84.

eite OB, Evensen O. Inflammatory cells of teleostean fish: a reviewfocusing on mast cells/eosinophilic granule cells and rodlet cells.Fish Shellfish Immunol 2006;20:192–208.

ombout JHWM, Van den Berg AA, van den Berg CTGA, Witte P,Egberts E. Immunological importance of the second gut segmentof carp. II. Characterization of mucosal lymphocytes. J Fish Biol1989;35:167–78.

oy PK, Munshi J.S.D. Malathion induced structural and morphome-tric changes of gills of a freshwater major carp Cirrhinus mrigala


(Ham). J Environ Biol 1991;12:79–87.aravana Bhavan P, Geraldine P. Histopathology of the hep-

atopancreas and gills of the prawn Macrobrachium malcolmsoniiexposed to endosulfan. Aquat Toxicol 2000;50:331–9.

PRESSemica xxx (2011) xxx–xxx

Schmale MC, Vicha D, Cacal SM. Degranulation of eosinophilicgranule cells in neurofibromas and gastrointestinal tract inthe bicolor damselfish. Fish Shellfish Immunol 2004;17:53–63.

Secombes CJ. The nonspecific immune system: cellular defences.In: Iwama G, Nakanishi T, editors. The fish immune system. SanDiego: Academic Press; 1996. p. 63–95.

Sharp GJE, Pike AW, Secombes CJ. The immune response ofwild rainbow trout Salmo gairdneri Richardson to naturallyacquired plerocercoid infections of Diphyllobothrium den-driticum (Nitzsch 1824) and D. ditremum (Creplin 1825). J FishBiol 1989;35:781–94.

Silphaduang U, Colorni A, Noga EJ. Evidence for widespread dis-tribution of piscidin antimicrobial peptides in teleost fish. DisAquat Organ 2006;72:241–52.

Silphaduang U, Noga EJ. Peptide antibiotics in mast cells of fish.Nature 2001;414:268–9.

Smith VJ, Fernandes MO. Antimicrobial peptides of the innateimmune system. In: Zaccone G, Meseguer J, Garcia-Ayala A,Kapoor BG, editors. Fish defenses, vol. 1: immunology. Enfield:Science Publishers; 2009. p. 241–75.

Sunitha S, Sahai S. Histopathological changes in the gills of Rasboradaniconius induced byg-BHC. J Environ Biol 1983;5:65–9.

Vallejo A, Ellis AE. Ultrastructural study of the response of


eosinophil granule cells to Aeromonas salmonicida extracel-lular products and histamine liberators in rainbow troutSalmo gairdneri Richardson. Dev Comp Immunol 1989;13:133–48.


mast cells in the intestine and gills of the sea bream, sparus aurata, exposed to a polychlorinated...

Documents