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

Neotropical Helminthology, 2018, 12(1), ene-jun:63-70.

ORIGINAL ARTICLE / ARTÍCULO ORIGINAL

INTRAMARSUPIAL DEVELOPMENT OF BRAGA PATAGONICA SCHIODT & MEINERT, 1884

(ISOPODA: CYMOTHOIDAE)

DESARROLLO INTRAMARSUPIAL DE BRAGA PATAGONICA SCHIODT & MEINERT, 1884

(ISOPODA: CYMOTHOIDAE)

atjsouza15@gmail.com*, mariliavmarilia@gmail.com, danielbporto81@gmail.com, jcmalta10@gmail.com

National Research Institute of the Amazon, Av. André Araújo, 2936 - Petrópolis, Manaus - AM, 69067-375, Brasil

ABSTRACT

The Intra marsupial developmental stages of Braga patagonica Schiodt & Meinert, 1884 collected from

fish in an Amazonian floodplain lake, were ascertained. Gravid females were screened according to their

gestational stage. Their eggs, embryos and larvae were measured and images were drawn. Six intra

marsupial stages have been identified, namely, eggs mass, egg, embryo differentiation I, embryo

differentiation II, Pullus I larva, Pullus II larva plus one Manca III post-marsupial infective swimmer-free.

Neotropical Helminthology

Amanda Karen Silva de Souza*; Marília Rodriguês da Vitória; Daniel Brito Porto & José Celso de Oliveira Malta

Keywords: Amazon – Fish Parasites – Peracarida – Phlabellifera

RESUMEN

Las etapas de desarrollo intramarsupiales de Braga patagonica Schiodt & Meinert, 1884 colectadas de

peces en un lago de planicie amazónica, fueron indagadas. Las hembras grávidas fueron seleccionadas de

acuerdo a su etapa de gestación. Sus huevos, embriones y larvas fueron medidos y dibujados. Seis estadios

intramarsupiales fueron identificados, a saber, la masa de huevo, el huevo, la diferenciación de embriones

I, diferenciación de embriones II, larva Pullus I, larva Pullus II más uno Manca III post- marsupial

infecciosa nadadora libre.

Palabras clave: Amazonas – parásitos de peces – Peracarida – Phlabellifera

Isopods are the most diverse and species-rich

crustaceans of the superorder Peracarida (isopods,

amphipods, tanaids, and their kin). Isopods are

common inhabitants of nearly all environments,

yet they are unusual among Crustacea due to their

ubiquity. Isopoda include approximately 10,000

described species, in 10 suborders. These animals

range in length from 0.5 mm to 500 mm.

Phylogenetic analyses and the fossil record

(limited though it is) suggest the group to date back

to at least the Paleozoic Carboniferous Period of

approximately 300 million years ago (Brusca et al.,

2001).

At the development of the cymothoid the eggs

show to be centrolecithal, oval and enclosed in a

chorion and vitelline membrane, the latter not

being clearly visible until the first few divisions

have taken place. The number of eggs carried in the

marsupium varies directly with body length within

a given species, ranging from about 200 to 1600. In

mouth/gill inhabiting species, the hatching mancas

always seem to be released via the host's gill

chamber and opercula opening (Holdich, 1968;

Brusca, 1981).

Manca and pullus larva stages typically bear large

eyes, spinose pereopods, and setose pleonal

appendages to facilitate swimming. Manca and

young males typically attach to almost any

convenient fish for short periods. Little is known

regarding these temporary attachments, although

some workers have referred to juveniles and young

males as “facultative parasites” on “intermediate

hosts”. It is not known whether the isopod actively

feeds on the fish at this time or not. As the juvenile

matures, it eventually finds an acceptable

“definitive” host fish upon which to attach to

permanently. The natatory setae are lost upon

attachment to this host and transition into a

functional male (Brusca, 1981).

The genus Braga Schiodte & Meinert (1881)

includes eight species: B. cichlae Schiodte &

Meinert (1881); B. nasuta Schiodte & Meinert

(1881); B. brasiliensis Schiodte & Meinert (1881);

B, patagonica Schiodte & Meinert (1884); B.

fluviatilis Richardson (1911); B. bachmanni

Stadler (1972); B. amapaenses Thatcher (1996)

Neotropical Helminthology, 2018, 12(1), ene-jun

INTRODUCTION

MATERIAL AND METHODS

and B. cigarra (Szidat & Shubart, 1960, Thatcher,

2002).

This work is the first one addressing a Braga

species' intra marsupial development and early

infectious stage.

Ten B. patagonica females were collected whilst

parasitizing the gills of Pygocentrus nattereri

(Kner, 1858), Serrassalmus elongatus (Kner,

1858), Brycon amazonicus (Spix & Agassiz, 1829),

Triportheus albus Cope, 1872 and Mylesinus sp.

The hosts were captured with gillnets at Catalão

0 0

Lake (03 10`04” S and 59 54`45”W). After being

captured each fish was examined in search for

Isopoda specimens. The isopods found were

collected, labeled and preserved in 70% ethyl

alcohol.

Females were measured and dissected, had their

eggs and larvae removed from the marsupium and

their development stage identified (Brusca, 1984;

Castro & Silva, 1985; Monteiro et al., 2001;

Ramdane et al., 2007).

Permanent preparations were made utilizing Hoyer

medium. Drawings were made with the aid of a

camera lucida. Measurements are in micrometers

(µm) unless designated as millimeters (mm). The

voucher material was deposited at the INPA

Crustacea Invertebrates Collection (INPA-2193,

2194, 2195, 2196 e 2197).

Ten females at different gravid stages were

analyzed. Six intramarsupial stages were

identified: eggs mass, egg, embryo I, embryo II

pullus I larva, and pullus II larva, as well as one

manca stage. At the intramarsupial development

stage, the analyzed females amounted to: one with

eggs mass, four with individualized eggs, one with

embryos at stage I, one with embryos at stage II,

one with pullus I larva stage and one with pullus II

RESULTS

Souza et al.

larva stage. In the post-marsupial stage one larva

was found at the manca stage.

Ten specimens of each stage and adult females

were measured in millimeters (Fig. 1).

Measurements averaged: Eggs 1.6 ± 0.1 long by

1.3 ± 0.1 wide; embryo I 1.7 ± 0.05 by 1.1 ± 0.03;

pullus I 2.7 ± 0.2 by 1.1 ± 0.04; pullus II 2.5 ± 0.2 by

1 ± 0.1; adults 17.4 ± 2 by 9.1 ± 1.5.

Intramarsupial Development

Stage 1 - Eggs mass (Fig. 2): initial stage of

embryonic development, grouping of the eggs,

oval in shape with no suspender filament. Each egg

is individualized in the structure's ensemble.

Stage 2 - Egg (Fig. 3): all eggs are morphologically

similar, circular-shaped and orange yellow colored

with lots of vitellus. With no segmentation. Total

number of eggs 88 (56-317). The egg size from four

females showed to be 19 ± 9.5 21 ± long by 11.8

wide.

Stage 3 - Embryo I (Fig. 4): head is differentiated

and initiates the differentiation of the eyes. The

differentiation of what will become the yolk sac

initiates in the dorsal part. The embryos are longer

than wide and 317 embryos I were found.

Stage 4 - Embryo II (Fig. 5): there is a very large

difference between head and body. Body is

Neotropical Helminthology, 2018, 12(1), ene-jun

differentiated in head, pereon, Pleon. The

rudiments of the telson, and gnathopod are visible.

Eyes well developed and visible. Yolk sac starts to

get circular-shaped, and 120 embryos II were

found. Female showed to be 1.9 ± 1.1 2.1 ± long by

1.1 wide.

Stage 5 - Larva pullus I (Fig. 6): head with

antennas, antennules and body segmentation quite

apparent. Pereon, Pleon, uropods and Telson

clearly differentiated. Six pairs of unornamented

legs. Visible yolk sac in the pereonite 3 dorsal part.

One exuvia externally coating the body, which had

undergone a molt when going to pullus II, was

observed six individuals were collected in the

marsupium.

Stage 6 - pullus II (Fig. 7): larger than the previous

stage, no exuvia was observed. The bristles and

thorns are evident on the pereopods. It has six pairs

of legs. All pullus II free in the marsupium. A total

of 196 pullus II were found. This is the last stage of

intramarsupial development, and is released into

the environment after molt as Manca I.

Post- marsupial development

Manca III (Fig. 8): the young individual is similar

to the adult male, with all the morphological

characters of the species but on a smaller scale. One

can already observe 7 pairs of legs and all the fully

formed body segments. Only one individual was

found parasitizing Triportheus albus Cope, 1872.

Intra marsupial development of Braga patagonica

Figure 1. Braga patagonica: Adult female.

Figures 2-5. intramarsupial early stages of development of B. patagonica. 1 - Egg Mass 2 - individual egg 3 - Side and back of

developing embryo I, a - eyes b- head; 4 - dorsolateral view of the developing embryo II.

Neotropical Helminthology, 2018, 12(1), ene-jun Souza et al.

2 3

4 5

Figure 6. Braga patagonica: Pullus I larva, a – legs, b - pereon, c - pleon, d – uropods, e - exuvia.

Neotropical Helminthology, 2018, 12(1), ene-jun Intra marsupial development of Braga patagonica

Figure 7. Braga patagonica: Pullus II larva.

Figure 8. Braga patagonica: Manca III larva.

Neotropical Helminthology, 2018, 12(1), ene-jun Souza et al.

DISCUSSION stage (Pugues et al., 1998).

The present work observed six intramarsupial

stages and one manca stage. The egg mass was

considered as the first intramarsupial development

stage according to Pugues et al. (1998). Some

researchers indicate the individualized egg as the

first stage (Alberto et al., 2001; Moreira, 1973;

Sartor & Pires, 1987).

In stage 3 the individualized eggs were more

elongated and the first visible differentiation under

protective membrane was the head and the visible

rudimentary eyes in dorsal and ventral view

(Alberto et al., 2001; Pugues et al., 1998). In this

work, stage 3 exhibited the same characteristics

and it was possible to observe the ocelos

primordium and the developing somatic mass.

Stage 4 of the marsupial development is marked by

the development of rudimentary eyes seen both in

dorsal and ventral view, the early formation of

appendages: antennae, pereiopods and pleopods

and the beginning of differentiation of a yolk sac,

which begins as a protuberance on the dorsal part

(Alberto et al., 2001; Moreira, 1973; Pugues et al.,

1998).

In this work we observed the same characteristics

mentioned by Moreira (1972), Pugues et al. (1998)

and Alberto et al. (2001). In the fourth B.

patagonica development stage the head, pereon

and pleon are well defined and there is the

beginning of the formation of appendages:

antennae, pereiopods and pleopods. The egg

membrane remains and the differentiation of the

yolk sac, which begins as a protrusion on the dorsal

part.

In stage 5 the eyes are already formed and the yolk

sac remains evident in A. trysibia (Pugue et al.,

1998). In T. henselii stage 5 the thoracic, abdominal

segments and six pairs of pereopods show to be

differentiated. The pereopods have neither

pigmentation nor ornamentation. The yolk sac is

located on the third pereonite. At this stage the

embryo is pullus I larva (Alberto et al., 2001).

The present work shows the B. patagonica stage 5

characteristics to be similar to those of the above

mentioned species. Braga patagonica differs by

having evident pigmentation at the ends and in the

The terminology used in the study of embryonic

development of isopods molts varies according to

the author. However, most of them point out an

intra and post-marsupial development period.

Inside the marsupium the eggs develop up to the

formation of the larvae. They leave maternal

oostegits with characteristics similar to those of the

male species, but lacking the seventh pair of

pereiopods that will only arise when they reach the

juvenile period following a succession of molts

(Alberto et al., 2001).

The post-larval and pubertal period is

characterized by a general growth of the young

individual, with the gradual emergence of the male

sexual characteristics. The pubertal period is

reached through a series of molts and it is

characterized in females and males by the

appearance of oostegits and sexual characteristics,

respectively (Alberto et al., 2001).

In B. patagonica there also occurred an intra and

post-marsupial development period. Stage I was

considered to be the egg mass not individualized in

th e m a r su p iu m . Fol l o w i n g th e eg gs

individualization differentiation of them will

initiate the development of subsequent

developmental stages.

In intramarsupial development of Telotha henselii

Von Martens, 1869 the sequence of embryo

segmentation stages with differentiation of

structures and the gradual reduction of yolk sac

(Alberto et al., 2001). In this work the

methodology adopted was that of Alberto et al.

(2001). Morphological differences were observed

in the development of B. patagonica indicating the

development to be similar between Isopoda species

with little variation from species to species.

In intramarsupial development stages of Livoneca

vulgares (Stimpson, 1857) the individualized eggs

developed themselves and there were five molts up

to the infective larva stage (manca) (Brusca, 1978).

Four intramarsupial development stages occurred

in Serolis complete (Moreira, 1971) (Albert et al.,

2001; Moreira, 1973; Sartor, 1987; Thatcher,

1988). In Artystone trysibia (Schiodte, 1866) the

egg mass was considered the first development

Alberto, RMF, Goulart, MS, Pereira, EHL. &

Bertoletti, JJ. 2001. Desenvolvimento

intramarsupial de Telotha henselii (Von

Martens, 1869) (Crustacea: Isopoda:

Cymothoidae). Comunicação do Museu de

Ciência e Tecnologia PUCRS série

Zoológica, vol. 14, pp. 73–87.

Brum, PED. & Araújo, PB. 2007. The manca

stages of Porcellio dilatatus Brandt

(Crustacea, Isopoda, Oniscidea). Revista

Brasileira de Zoologia ,vol. 24, pp. 93–502.

Brusca, RC. 1978. Studies on the cymothoid fish

symbionts of the eastern Pacific

(Crustacea: Isopoda: Cymothoidae), 2:

Biology and systematics of Lironeca

vulgaris. Allan Hancock College

Foundation, Occasional paper vol. 2, pp.

1–19.

Brusca, RC. 1981. A monograph on the Isopoda

Cymothoidae (Crustacea) of the eastern

Pacific. Zoological Journal of the Linnean

Society, vol. 73, pp. 117–199.

Brusca, RC. 1984. Phylogeny, evolution and

biogeography of the marine isopod

subfamily Idoteinae (Crustacea: Isopoda:

Idoteidae). Transactions of San Diego

Society of Natural History, vol. 20, pp.

99–134

Castro, AL & Silva, JL. 1985. Manual de

identificação de invertebrados límnicos do

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Holdich, DM. 1968. Reproduction, growth and

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BIBLIOGRAPHIC REFERENCES

Neotropical Helminthology, 2018, 12(1), ene-jun Intra marsupial development of Braga patagonica

middle of pereonites and pleonites; they have no

ornamentation and there is a cuticle surrounding

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Stage 6 is called pullus II and in T. hanselii the

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uropods are evident. The seventh pair of

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II was also the last stage within the marsupium.

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work with B. patagonica the manca stage was

attached to mouth of T. albus probably in stage 3

development. T. albus may likely exhibit the

mancas' latest stages.

Received February 23, 2018.

Accepted April 3, 2018.

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