Aquaculture Reports 2 (2015) 22–25

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Esophageal infection due to Kudoa sp. (Myxozoa) in mapara catfish,

Hypophthalmus marginatus

a b a c

Michele Velasco , Marcela Videira , José Mauro Viana da Silva , Osimar Sanches ,

d e a,∗

Patrícia Santos Matos , Sérgio Carmona de São Clemente , Edilson Matos

a

Carlos Azevedo Research Laboratory, Federal Rural University of Amazonia (UFRA), Belém, Pará, Brazil

b

Morphophysiology and Health Laboratory, Amapá State University (UEAP), Macapá, Amapá, Brazil

c

MV. Pathologist – Veterinary Diagnostic Center – CDAPVET, Presidente Prudente, São Paulo, Brazil

d

Edilson Matos Research Laboratory, Federal University of Pará (UFPA), Belém, Pará, Brazil

e

Faculty of Veterinary, Federal Fluminense University (UFF), Niterói, Rio de Janeiro, Brazil

a

r t i c l e i n f o a b s t r a c t

Article history: The present study describes lesions caused by Kudoa sp. in the esophageal tissue of mapará catfishes

Received 26 November 2014

( marginatus) collected from Cametá, Tocantins River in the northern Brazil. Fifty refriger-

Received in revised form 23 April 2015

ated H. marginatus specimens were dissected and tissue samples were for embedded in paraffin and

Accepted 1 May 2015

stained with hematoxylin-eosi, Masson’s trichrome and Giemsa. Pseudocysts of whitish color were

Available online 22 May 2015

observed in the esophagus region. The parasites had four valves and other morphological characteristics

of the genus Kudoa. Microscopic analysis revealed lesions in the mucosa, submucosa, and muscular layers.

Keywords:

The Masson staining highlighted the presence of myxosporean cysts in the cytoplasm of the muscular

Amazonia

Esophagus fibers, and marked fibrosis of the muscular and mucous layers. The macro and microscopic findings of

Disease the present study confirm the presence of necrotic esophagitis in H. marginatus associated with infection

Mapará by Kudoa.

Myxozoan © 2015 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND

license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

1. Introduction As result of the herbivorous habit, high filet yield, good meat

taste and nutritional standards (Costa et al., 2010), associated to

The Amazon has the greatest diversity of freshwater fish in the great commercial importance, H. marginatus has been studied

the world (Santos and Santos, 2005). However, there is a lack for farming. Recently, the was included in lists of fish with

of knowledge on the ichthyofauna inhabiting the various aquatic potential for aquaculture, divulged by govern agencies and private

environments available (Soares et al., 2008). The high diversity of companies. (Brasil, 2015). In this context, know the parasites find in

habitats creates conditions for evolving many fish species with natural populations is important to prevent potential infestations

different characteristics. Some of them have great potential for in culture conditions.

aquaculture and high value on the international market, such as The importance of ichthyoparasites has increased in the past

the mapará catfish Hypophthalmus marginatus Valenciennes, 1840. years because of their implications for human health and economic

It is a peculiar siluriform, which is pelagic and phytoplanktopha- activities. Parasitological studies on H. marginatus are rare in the

gus. Mapara is largely exploited by fisheries (Worthmann, 1980) literature. The myxosporidiosis in this species is more important.

and most of the catch is exported abroad or to other regions in Recently, Rocha et al. (2014) described a new species of myx-

Brazil (Cutrim and Batista, 2005). osporean, Thelohanellus marginatus, infecting gills of mapara. A

number of myxosporean species, especially those of the genus

Kudoa Meglitsch, 1947, are of considerable importance due to their

∗ effects on the quality of fishery products, in particular due to post-

Corresponding author at: Laboratório de Pesquisa Carlos Azevedo, Universidade

◦ mortem myoliquefaction of the muscle tissue, which is rendered

Federal Rural da Amazônia, Avenida Presidente Tancredo Neves, N 2501 Bairro,

Montese, Cidade, Belém, Pará Cep: 66.077-901, Brazil. Tel.: +55 913210 5207. unacceptable for human consumption (Morado and Sparks, 1986;

E-mail addresses: michele [email protected] (M. Velasco), Stehr and Whitaker, 1986; Moran et al., 1999a). The genus Myxobo-

[email protected] (M. Videira), [email protected]

lus are also described as a causative of myoliquefaction of in fish.

(J.M. Viana da Silva), [email protected] (O. Sanches), [email protected]

In south Brazil, Eiras et al. (2005) have reported myoliquefac-

(P. Santos Matos), [email protected] (S. Carmona de São Clemente),

tion in the flounder Paralichthys orbignyanus due to infection by

[email protected] (E. Matos).

http://dx.doi.org/10.1016/j.aqrep.2015.05.001

2352-5134/© 2015 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4. 0/).

M. Velasco et al. / Aquaculture Reports 2 (2015) 22–25 23

Fig. 1. Hypophthalmus marginatus. (A) Macroscopic view of the whitish cysts in the esophagus region (*). (B) Optical photomicrographs by means of differential interference

contrast (DIC; Nomarski) showing in natura Kudoa spores in apical view (Ep). Scale bar 10 ␮m.

Fig. 2. Hypophthalmus marginatus. (C) Transverse section of the esophagus region, showing edema in the submucosa (arrowheads) and necrosis of the muscle area (arrow). HxE.

Scale bar 100 ␮m. (D) Mucosa of the esophagus region infiltrated by lymphocytes (arrow). HxE. Scale bar 100 ␮m. (E) Transverse section of the esophagus region, showing

pseudocysts of Kudoa in the muscle layer (Ci) and presence of a large quantity of free spores in the mucosa region (arrow). Giemsa. Scale bar. 100 ␮m. (F) Observation

intracitoplasmatic pseudocysts Kudoa sp. (arrows), and necrosis of the muscle fibers (arrowheads). T. Masson. Scale bar. 100 ␮m. (G) Fibrosis in the mucosa and submucosa

(arrows) and intense infiltration of lymphocytes (arrowheads). Masson T. Scale bar 100 ␮m. (H) Muscle layer of the esophagus, showing fibrosis surrounding the muscle

fibers (arrows) and cysts containing Kudoa spores (arrowheads). Masson T. Scale bar 100 ␮m.

24 M. Velasco et al. / Aquaculture Reports 2 (2015) 22–25

Myxobolus sp. This muscle lysis result from action by proteases regions. Moran et al. (1999a) observed that when spores were

produced parasites, resulting in myoliquefaction in the tissue of released from pseudocysts through bursting, they generated an

their host. (Moran et al., 1999b). The present study describes inflammatory reaction with phagocyte involvement. Other stud-

macroscopic and microscopic lesions caused by Kudoa cysts in the ies, such as those by Andrada et al. (2005) on Trichiurus lepturus,

esophageal region of the mapará catfish H. marginatus in the Ama- Heckmann and Jensen (1978) on Sebastes paucispinis and Casal

zon basin, northern Brazil. et al. (2008) on Aequidens plagiozonatus, described infections due to

Kudoa with little or no inflammatory reaction. According to those

findings, infection of the esophageal musculature of H. margina-

2. Material and methods

tus by Kudoa sp. caused damage to the host fish, culminating in

Fifty specimens of the mapara H. marginatus, of 30 cm length inflammatory process and of fibrosis in the muscle layer, due to

(28–34 cm) and 75 g weight (72–78 g), were acquired dead from intense infection of the muscle fiber, and in the submucosa layers

◦ ◦

artisan fishermen near the town of Cametá (2 14 S, 49 29 W), in and mucosa of the esophagus, due to bursting of the pseudocysts

the northern Brazil state of Pará, between July 2011 and May 2012. with spore releasing.

The specimens were transported in refrigerated containers to the In aquaculture, the fish are subject to infection by numerous

Carlos Azevedo Research Laboratory at the Federal Rural University species of parasites, due to various stress factors provided by the

of Amazonia, in Belém. The fish were necropsied and the esophageal management (Tavares-Dias et al., 2001). Then, the results obtained

portion of the gastrointestinal tract was observed under a stereomi- in this work are fundamental to characterize the parasitological

croscope appropriate for parasitological studies. profile of the H. marginatus. It may subsidize research to standardize

For fresh examination, small tissue fragments with suspected methods of control and prevention, and avoid spread diseases in

parasitism were placed between slides and coverslips with a drop in aquaculture.

of water, for observation under an optical microscope and a dif-

ferential interference contrast (DIC) microscope (Nomarski). For Acknowledgements

the histological procedure, small fragments (0.5 cm) of the par-

asitized tissue extracted from the esophageal region of the fish We are grateful to Coordenac¸ ão de Aperfeic¸ oamento de Pessoal

specimens were fixed in Davidson’s solution (neutral-buffered for- de Nível Superior (CAPES), Conselho Nacional de Desenvolvimento

malin, glacial acetic acid, 95% ethanol and distilled water) for 24 h Científico e Tecnológico (CNPq), Fundac¸ ão Amazônia Paraense de

and processed for setting in paraffin and stained with Masson’s Amparo à Pesquisa (FAPESPA) and The Edilson Matos Research Lab-

trichrome, hematoxylin-eosin and Giemsa (Luna, 1968). The sec- oratory (LPEM–UFPA) and to Mr. David Elliff for idiom revision

tions were stained and mounted on slides with a coverslip and of the manuscript. The helpful suggestions and comments of the

analyzed in a Nikon E-200 light microscope equipped with the Associate Editor and reviewers are greatly appreciated.

®

Micrometrics SE Premium image-capturing software.

References

3. Results and discussion

Andrada, C.D.G., Tortelly, R., Nogueira, P.P., Andrade, C.L., Lima, F.C., 2005. Infecc¸ ão

por Kudoa Meglitsch, 1947 (Myxozoa: Multivalvulida) em musculatura

The necropsy on H. marginatus specimens revealed that 60% of

esquelética de espada Trichiurus lepturus L. (Teleostei: Trichiuridae). Parasitol.

the hosts (30/50) presented pseudocysts of whitish color and fila-

Latinoam. 60, 150–153, http://dx.doi.org/10.4067/S0717-77122005000200008

mentous shape, in the region of the esophagus (Fig. 1A). From fresh Brasil, Ministério da Pesca e Aquicultura Secretária de Estado de Pesca e

Aquicultura. Pará, 2015. Quadro Sinótico de Espécies de Peixes Potenciais para

examination under the microscope, it was observed the mature

Piscicultura. Available in: www.sepaq.pa.gov.br

spores exhibiting the diagnostic traits of spores of the genus Kudoa

Bunton, T.E., Poynton, S.L., 1991. Kudoa sp. (Myxosporea: Multivalvulida) infection

(Fig. 1B): radial symmetry composed of four valves, with one polar in juvenile white perch, Morone Americana (Gmehn): histopathology and spore

morphology. J. Fish Dis. 14, 589–594, http://dx.doi.org/10.1111/j. 1365-2761.

capsule in each valve, as described by Moran et al. (1999b) and Lom

1991. tb00615.x

and Dyková (2006).

Casal, G., Matos, E., Matos, P., Azevedo, C., 2008. Ultrastructural description of a

The histopathological analysis on the fragments of parasitized new myxosporean parasite Kudoa aequidens sp. n. (Myxozoa, Myxosporea)

tissue revealed multifocal necrosis of the esophageal epithelium, found in the sub-opercular musculature of Aequidens plagiozonatus (Teleostei)

from the Amazon River. Acta Protozool. 47, 135–141.

with marked edema of the submucosa (Fig. 2C) and inflammatory

Costa, T.V., Oshiro, L.M.Y., Silva, E.C.S., 2010. O potencial do mapará Hypophthalmus

infiltration composed by lymphocytes and macrophages, surround-

spp. (Osteichthyes, Siluriformes) como uma espécie alternativa para a

ing the epithelial cells associated with the necrotic tissue (Fig. 2D). piscicultura na Amazônia. Bol. Inst. Pesca. 36 (3), 165–174.

Cutrim, L., Batista, V.S., 2005. Determinac¸ ão de idade e crescimento do mapará

The muscle layer was characterized by severe coagulative necro-

(Hypophthalmus marginatus) na Amazônia Central. Acta Amazon 35 (1), 85–92,

sis of the striated muscle fibers and an abundance of pseudocysts http://dx.doi.org/10.1590/S0044-59672005000100013

containing Kudoa spores, in the cytoplasm of the muscle fibers Eiras, J.C., Júnior, J.P., Sampaio, L.A., Robaldo, R., Abreu, P.C., 2005. Myxobolus sp. can

cause in vivo myoliquefaction in the host Paralichthys orbignyanus

(Fig. 2E and F). Similar data were described by Stehr and Whitaker

(Osteichthyes, Paralichthydae). Dis. Aquat. Org. 77, 255–258, http://dx.doi.org/

(1986) and Morado and Sparks (1986) in the musculature of the 10.3354/dao01852

host Merluccius productus infected with Kudoa thyrsitis and Kudoa Heckmann, R.A., Jensen, L.A., 1978. The histopathology and prevalence of

Henneguya sebasta and Kudoa clupeidae in the rockfish, Sebastes paucispinis of

paniformis, by Bunton and Poynton (1991) in the musculature of

Southern California. J. Wildlife Dis. 14, 259–262.

Morone Americana and by Eiras et al. (2005) in the musculature of

Lom, J., Dyková, I., 2006. Myxozoan genera: definition and notes on ,

the fish P. orbignyanus infected with Myxobolus sp. life-cycle terminology and pathogenic species. FoliaParasitologica 53, 1–36.

Luna, L.G., 1968. Manual of Histologic Staining Methods of the Armed Forces

Masson’s trichrome stained myxosporean cysts in the cytoplasm

Institute of Pathology, 3rd ed. MacGraw-Hill Book Company, New York, pp.

of the muscle fibers, revealed fibrosis of the muscle layer and the

251.

mucosa (Fig. 2G and H). Morado and Sparks (1986) reported that Morado, J.F., Sparks, A., 1986. Observations on the host-parasite relations of the

encapsulation of the plasmodia of K. thyrsitis & K. paniformis took Pacific whiting, Merluccius productus (Ayres), and two myxosporean parasites,

Kudoa thyrsitis (Gilchrist, 1924) and K. paniformis Kabata & Whitaker, 1981. J.

place through development of plasmodia over the entire extent of

Fish Dis. 9, 445–455.

the host’s muscle fiber, starting with a phagocytic infiltrate and

Moran, J.D.W., Margolis, L., Webster, J.M., Kent, M.L., 1999a. Development of Kudoa

progressing to encapsulation by fibroblasts. thyrsites (Myxozoa: Myxosporea) in netpen-reared Atlantic salmon

determined by light microscopy and a polymerase chain reaction test. Dis.

A number of burst pseudocysts could also be observed, with

Aquat. Org. 37, 185–193.

spores disseminated in the cells of the muscle layer, submucosa

Moran, J., D,W, Whitaker, D.J., Kent, M.L., 1999b. A review of the myxosporean

and lumen of the esophagus, with observation of fibrosis in these genus Kudoa Meglitsch, 1947, and its impact on the international aquaculture

M. Velasco et al. / Aquaculture Reports 2 (2015) 22–25 25

industry and commercial fisheries. Aquaculture 172, 163–196, http://dx.doi. Stehr, C., Whitaker, D.J., 1986. Host-parasite interaction of the myxosporeans

org/10.1016/S0044-8486(98)00437-2 Kudoa paniformis Kabata & Whitaker, 1981 and Kudoa thyrsites (Gilchrist, 1924)

Rocha, S., Casal, G., Velasco, M., Alves, A., Matos, E., Al-Quraishy, S., Azevedo, C., in the muscle of Pacific whiting, Merluccius productus (Ayres): an

2014. Morphology and phylogeny of Thelohanellus marginatus n. sp. (Myxozoa: ultrastructural study. J. Fish Dis. 9, 505–517.

Myxosporea), a parasite infecting the gills of the fish Hypophthalmus Tavares-Dias, M., Martins, M.L., Moraes, F.R., 2001. Fauna parasitária de peixes

marginatus (Teleostei: ) in the Amazon River. J. Eukaryot. oriundos de pesque-pague do município de Franca, São Paulo, Brasil I.

Microbiol., 586–593. Protozoários. Revta Bras. Zool. 18 (1), 67–79.

Santos, G.M., Santos, A.C.M., 2005. Sustentabilidade da pesca na Amazônia. Estud. Worthmann, H., 1980. Estágios iniciais de crescimento da pescada (Plagioscion

Av. 19 (54), 165–182. monti Soares). Acta Amazon 10 (1), 157–163.

Soares, E.C., Teixeira, C.V., Oliveira, A.C., Parise, M., Pinto, W.H.A., 2008. Avaliac¸ ão

da pesca através do banco de estatística e SIG na região de Santarém, estado do

Pará. Brasil Rev. Bras. Eng. Pes. 3 (1), 98–107.