RESEARCH NOTE/NOTA CIENTÍFICA

FIRST RECORD OF ANISAKIS SP. (NEMATODA, ANISAKIDAE) L3 INFECTING THE

BODY CAVITY OF ATLANTORAJA PLATANA (CHONDRICHTHYES, RAJIDAE)

PRIMER REGISTRO DE ANISAKIS SP. (NEMATODA, ANISAKIDAE) L3 EN LA

CAVIDAD CORPORAL DE ATLANTORAJA PLATANA (CHONDRICHTHYES, RAJIDAE)

1 2 4 1 2 4 5 2 3

A.C. Moya , E.J Galíndez , E.E. Di Giacomo & R.D. Tanzola

1 Laboratorio de Citología, Histología y Embriología Animal, DBByF, UNS, San Juan 670, 8000, Bahía Blanca, Argentina.

2 Instituto de Investigaciones Biológicas y Biomédicas del Sur (INBIOSUR)-CONICET (Consejo Nacional de

Investigaciones Científicas y Tecnológicas).

3 Laboratorio de Patología de Organismos Acuáticos, DBByF, UNS, San Juan 670, 8000, Bahía Blanca, Argentina.

4 CONDROS-Laboratorio de Recursos Ícticos. Instituto de Biología Marina y Pesquera “Alte. Storni”, Güemes 1030,

CP8520, San Antonio Oeste, Río Negro.

5 Universidad Nacional del Comahue, San Martín 247, CP 8520, San Antonio Oeste, Río Negro, Argentina. Tel/Fax: 54 –

2934 – 21002.

Corresponding-authors: acmoya83@gmail.com ; rtanzola@uns.edu.ar

Neotropical Helminthology, 2015, 9(2), jul-dec: 359-365.

ABSTRACT

Keywords: Anisakis sp. - Atlantoraja platina - Epigonal organ - L - Nematodes.

This communication is the first record of the presence of a third stage larva of Anisakis sp.

infecting Atlantoraja platana. The hosts were collected from fishery landings at processing

plants of San Antonio Oeste (40° 44' S 64° 57' O) and San Antonio Este ports (40° 49' S 64° 57'

O), Rio Negro province, Argentina. They were found in the visceral cavity near the epigonal

organ, a lymphomyeloid tissue closely associated with gonads and only in cartilaginous fish. The

high concentrations of urea in the body fluid and tissues of elasmobranch hosts made an

inhospitable environment to the colonization of helminthes. The results produced by this work

constitute the first report of L of Anisakis sp. in the body cavity of an elasmobranch, in particular

A. platana, and show the capability of this anisakid to survive in the visceral mass of these hosts.

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ISSN Versión impresa 2218-6425 ISSN Versión Electrónica 1995-1043

RESUMEN

Palabras clave: Anisakis sp. L - Atlantoraja platana - Epigonal organ - Nematodes.

En la presente nota se registra por primera vez el tercer estadio larval de Anisakis sp. infectando a

Atlantoraja platana (Günther, 1880). Los hospederos fueron obtenidos en desembarques

pesqueros de plantas de procesado de los puertos de San Antonio Oeste (40° 44' S 64° 57' O) y San

Antonio Este (40° 49' S 64° 57' O), Provincia de Río Negro, Argentina. Las larvas fueron

colectadas en la cavidad visceral de los peces, cerca del órgano epigonal, un tejido linfomieloide

estrechamente asociado a las gónadas y exclusivo de los peces cartilaginosos. Hay evidencias

documentadas que las altas concentraciones de urea en tejidos y fluídos corporales tornan

inhabitable el medioambiente celómico para ser colonizado por helmintos. Los resultados

expuestos en este trabajo constituyen el primer reporte de L de Anisakis sp. en la cavidad corporal

de un elasmobranquio, en particular A. platana, y demuestran la capacidad de este anisákido para

sobrevivir en la masa visceral de estos hospederos.

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INTRODUCTION

The anisakid nematodes parasitize

elasmobranches as larvae or adults and are

generally internal parasites (Caira, 1990)

besides they can be found at integumentary

level. The location where larval nematodes can

be found range from the uterus (Benz et al.,

1987) and ovaries (Rosa-Molinar et al., 1983;

Aragort et al., 2002) to superficial body tissues

(Ruyck & Chabaud, 1960).

Anisakids use fish as paratenic hosts to close

their complex life cycles (Guagliardo et al.,

2009). There are relatively few records of

nematodiasis in rays (Mc Vicar 1977; Romera,

1993; Tanzola et al., 1998; Borucinska &

Heger, 1999; Sanmartín et al., 2000, Knoff et

al., 2001; Aragort et al., 2002; Santos et al.,

2004; Alvarez et al., 2006; Díaz Andrade et al.,

2008). The present communication is the first

report of the third stage larva (L ) of Anisakis

sp. infecting the body cavity of Atlantoraja

platana (Günther, 1880). This skate is an

endemic species from the Southwest Atlantic

Ocean and one of the most commonly taken as

bycatch in the coastal fisheries at the San

Matías Gulf (Northern Patagonia, Argentina).

Its IUCN Red List Category is “Vulnerable

A4bd”(http://www.iucnredlist.org/details/631

10/0 ).

Males specimens of A. platana were collected

from fishery landings at processing plants of

San Antonio Oeste (40° 44' S 64° 57' O) and

San Antonio Este ports (40° 49' S 64° 57' O),

Rio Negro province, Argentina. The specimens

were taxonomically identified following

Coller (2012). Two of fifteen (13.3%

prevalence) result parasitized by anisakid

larvae. These findings proceed from

histopathological examination.

Small pieces of the male reproductive system

were fixed in Bouin's solution in seawater for

at least 24 h. Afterwards, all material was

dehydrated through a graded series of ethanol

and embedded in Paraplast®. Sections of 4-5

μm thick were stained by Masson's trichromic

stain and hematoxilyn-eosin. Selected sections

were photographed using an Olympus BX51

light microscope equipped with an Olympus

C-7070 digital camera.

Following the criteria of Oshima (1972) the

larvae were identified as third stage (L ) of

Anisakis sp. and were characterized by a

cuticle with slight ripples, number of muscular

cells (41-56 muscular cells per quadrant) and

hypodermic cords with “Y-shape” associated

with the excretory gland (Figs. 1 y 2). The

maximum diameter of the larvae were between

0.35 – 0.37 mm.

The parasites were found in visceral cavity of

two males of A. platana (one juvenile and other

sexually mature). In the juvenile specimen,

larvae were found near the epigonal tissue

adjacent to the testes. In this case,

inflammatory tissue was evident (Fig. 3). In

the mature host, larvae were found near the

epigonal organ bordering the genital ducts and

no inflammatory tissue was detected (Fig. 4).

Despite the presence of larvae, the

cytoarchitecture of reproductive and

lymphomyeloid structures of the hosts was

preserved.

The larvae stages of Anisakis sp. present in fish

tissues can be determined, in cross section, by

the structure of the digestive tract, the

morphology of the hypodermic cords and the

association of these structures with the

excretory gland (Oshima, 1972). However,

few characters of the adults are present in

larvae, so the specific determination of the

MATERIAL AND METHODS

Moyaet al.

RESULTS AND DISCUSSION

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parasite, based on the larvae, is complex (Dick

& Choudhury, 1995; Guagliardo et al., 2009).

The general pathology of fish specify that the

major diseases caused by nematodes have

targeted organs as intestine, liver, heart, body

cavity, muscle, gonads (especially ovary) and

swim bladder (Williams & Jones, 1994; Dick

& Choudhury, 1995). Borucinska & Heger

(1999) associated the presence of granulomas

scattered throughout different organs (spleen,

stomach, spiral valve, kidney and gill septa) in

Isurus oxyrinchus (Rafinesque, 1891) with

nematoda larvae and were attributed likely to

drancunculoids.

Santos et al. (2004) reported the occurrence of

a P s e u d o a n i s a k i s ( A s c a r i d i d a :

Acanthocheilidae) from the intestines of two

rajids: Rioraja agassizii (Müller & Henle,

1841) and Psammobatis extenta (Garman,

1913) from Brazilian southwestern Atlantic

waters. Alvarez et al. (2006) recorded six

species of nematode, among them A. simplex,

all of them in the lumen of stomach and

intestine.

Knoff et al., (2001) reported L of Anisakis sp.

in the stomach and spiral valve of 263

elasmobranchs of Brazil.

Guagliardo et al. (2009) found L of Anisakis

sp. in a bony fish Seriolella porosa (Guichenot,

1848) and Tanzola & Guagliardo (2004)

reported L of several anisakids in the intestinal

lumen of cartilaginous fish, but no invasion of

these in the body cavity. The Anisakis larvae

found in the present study are morphologically

similar and share the range of measurements

with those parasitize silver warehou, S.

porosa, from the same geographical area, San

Matias Gulf (Guagliardo et al., 2009).

Anisakis sp. l infecting the body cavity of Atlantoraja platana

Figure1. High magnification of the anterior part of a third larva stage of Anisakis sp. Arrow shows the cuticle. Asterisks indicate

the hypodermic cordons. Mc: muscular cells; Oe: oesophagus; Eg: excretory gland. Scale bar: 70 µm.

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The body cavity of elasmobranch is rarely

parasitized by nematodes (Caira & Healy,

2004). Moravec & Little (1988) reported two

species of micropleurid nematodes in bull

sharks and Caira & Healy (2004) found larvae

of ascarid nematodes in elasmobranchs from

the Gulf of California. Williams (1964) and

Williams et al. (1970) stated that the high

concentrations of urea in the body fluid and

tissues of elasmobranch hosts made an

inhospitable environment to the colonization

of helminthes, such as acanthocephalans. This

statement could be extrapolate to explain the

few records of larval helminths parasitizing the

body cavity and tissues of cartilaginous fishes

to date (Díaz Andrade et al., 2008).

Base on this background, the results exposed in

this work constitute the first report of L of

Anisakis sp. in the body cavity of an

elasmobranch, in particular A. platana, and

show the capability of this anisakid to survive

in the visceral mass of these hosts.

Moyaet al.

Figure2. High magnification of the middle part of a third larva stage of Anisakis sp. Asterisks indicate hypodermic cordons. In:

intestine. Scale bar: 60 µm.

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Anisakis sp. l infecting the body cavity of Atlantoraja platana

Figure3. Transverse section of a third larva stage of Anisakis sp. in visceral tissue of a juvenile male of Atlantoraja platana.

Asterisk depicts inflammatory tissue. Ct: connective tissue. Scale bar: 130 µm.

Figure4. General view of a third larval stage of Anisakis sp. in visceral tissue of a mature male of A. platana. Eo: epigonal organ;

Ct: connective tissue; Gd: genital ducts. Scale bar: 150 µm.

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ACKNOWLEDGEMENT

We thank to San Antonio Oeste and San

Antonio Este fish-processing plants for

providing us the samples. This work was

supported by the SGCyT-UNS, PGI: 24/B173.

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Anisakis sp. l infecting the body cavity of Atlantoraja platana

Received June 29, 2015.

Accepted September 26, 2015.