The Kumamoto oyster, Crassostrea sikamea, starts gametogenesis as young as 71 days old from spawning (35 days from postsettlement) with a mean shell height of 3.0 mm. This information constitutes a new record in age-size for gametogenesis in oysters for commercial importance and adds another biological difference comparing this species with the Pacific oyster Crassostrea gigas

Cáceres-Martínez, Jorge

Guerrero-Rentería, Yanet

Vásquez-Yeomans, Rebeca

Spanish

Hidrobiológica (E-Journal)

Notas	 181Vol.	22	No.	2	•	2012Early Gametogenesis of Kumamoto oyster (Crassostrea sikamea )Gametogénesis temprana en el ostión Kumamoto (Crasssotrea sikamea )Jorge	Cáceres-Martínez,1,2	Rebeca	Vásquez-Yeomans2	and	Yanet	Guerrero-Rentería11 Centro de Investigación Científica y de Educación Superior de Ensenada. Departamento de Acuicultura. Carretera Ensenada-Tijuana No. 3918, Zona Playitas. Ensenada, Baja California, 22860. México 2 Instituto de Sanidad Acuícola, A. C. (ISA). Calle 15 #265, entre Obregón y Moctezuma, Zona Centro. Ensenada, Baja California, 22800. México e-mail: jcaceres@cicese.mxCáceres-Martínez	J.,	R.	Vásquez-Yeomans	and	Y.	Guerrero-Rentería. 2012.	Early	Gametogenesis	of	Kumamoto	oyster (Crassostrea sikamea).	Hidrobiológica	22(2):	181-184.ABSTRACTThe	 Kumamoto	 oyster,	 Crassostrea sikamea,	 starts	 gametogen-esis	as	young	as	71	days	old	from	spawning	(35	days	from	post-settlement)	with	a	mean	shell	height	of	3.0	mm.	This	information	constitutes	a	new	record	in	age-size	for	gametogenesis	in	oysters	for	 commercial	 importance	 and	 adds	 another	 biological	 differ-ence	comparing	this	species	with	the	Pacific	oyster	Crassostrea gigas.Key words:	Crassostrea sikamea,	gametogenesis,	gonadal	devel-opment,	Kumamoto	oyster,	sexual	differentiation.RESUMENEl	ostión	Kumamoto,	Crassostrea sikamea,	inicia	la	gametogéne-sis	a	una	edad	de	71	días	después	del	desove	(35	días	después	del	asentamiento)	con	un	promedio	de	altura	de	la	concha	de	3.0	mm.	Esta	información	constituye	un	nuevo	registro	en	la	edad	y	talla	en	 la	 cual	 se	 inicia	 la	 gametogénesis	 en	 ostiones	 de	 importan-cia	comercial.	Adicionalmente,	pone	en	evidencia	otra	diferencia	biológica	respecto	al	ostión	Japonés	Crassostrea gigas.Palabras clave:	 Crassostrea sikamea,	 desarrollo	 gonadal,	 dife-renciación	sexual,	gametogénesis,	ostión	Kumamoto.The	Kumamoto	oyster,	Crassostrea sikamea	Amemiya,	1928,	was	introduced	for	commercial	culture	into	bahía	de	San	Quintín,	Baja	California,	Mexico	around	1970	from	the	West	coast	of	the	U.S.A.	as	 a	 variety	 of	 the	 Pacific	 oyster	 Crassostrea gigas	 (Thunberg,	1793)	 (Gobierno	del	Estado	de	Baja	California	Sur,	2010).	Nowa-days,	it	is	known	that	the	Kumamoto	oyster	is	a	separate	species	from	the	Pacific	oyster	based	on	molecular	characterization,	mi-crosatellite	 DNA	 markers,	 concordant	 differences	 in	 16S	 rDNA	and	 allozymes	 (Banks	et al.,	 1993,	 1994;	 Hedgecock	 et al.,	 1999;	Sekino	et al.,	2003;	Reece	et al.,	2008;	López-Flores	et al.,	2010).	Additionally,	 there	are	phenotypic	and	physiological	differences	between	 C. gigas	 and	 C. sikamea,	 the	 later	 has	 slower	 growth	rate,	 smaller	 size,	 smaller	 eggs,	 a	 more	 deeply	 cup-shaped	 left	valve,	and	a	highly	wrinkled	or	ridged	shell	(Amemiya,	1928;	Nu-machi,	1978).	The	age	and	size	for	gametogenesis	could	also	be	different	among	genetically	related	species.	The	smaller	size	for	gametogenesis	in	C. gigas	is	between	16	to	39	mm	of	shell	height	(Buroker,	1983),	while	for	C. virginica	 (Gmelin,	1791)	 it	 is	smaller	than	35	mm	of	shell	height,	and	as	young	as	42	days	from	post-settlement	 (Stafford,	1913;	Thompson	et al.,	 1996).	The	oyster	C. rizophorae	(Guilding,	1828)	begins	gametogenesis	at	the	small-est	sizes;	from	5	to	9	mm	of	shell	height	and	less	than	42	days	old	from	post-settlement	(Vélez,	1976).	Until	now,	there	is	no	informa-tion	about	the	age	and	size	for	the	beginning	of	gametogenesis	in	C. sikamea.	During	a	histological	survey	of	Kumamoto	oysters	of	<	4.5	mm	of	shell	height	and	35	days	old	from	post-settlement,	sur-prisingly,	we	observed	gametogenesis.	Results	 from	this	survey	are	described.In	August	2007,	one	sample	of	120	Kumamoto	oysters	 from	a	commercial	hatchery	 in	Sinaloa,	Northwest	Mexico,	was	sent	alive	to	the	Laboratory	of	the	Instituto	de	Sanidad	Acuícola,	A.C.	for	 histological	 analysis.	 The	 sample	 was	 composed	 by	 oysters	of	71	days	old	from	spawning	and	35	days	from	post-settlement,	cultured	at	24	oC.	A	second	sample	of	80	oysters	of	similar	age	and	cultured	at	the	same	temperature	was	sent	a	month	later	to	the	same	laboratory.	The	oysters	were	cleaned	using	running	water	Hidrobiológica			2012,	22	(2):	181-184	 Agosto	2012NOTAS182	 Notas	 Hidrobiológicato	eliminate	debris.	From	each	sample,	54	and	58	organisms	were	chosen	 respectively	 due	 to	 best	 appearance	 and	 entire	 shells.	These	 organisms	 were	 considered	 as	 subsamples	 and	 were	measured	 using	 a	 micrometer	 placed	 in	 a	 stereo	 microscope,	considering	the	shell	height	(distance	from	the	umbo	to	the	distal	posterior	border	of	the	shell;	Helm	et al.,	2006).	The	whole	oysters	were	fixed	by	24	h	in	Davidson’s	fixative	solution	(Shaw	&	Battle,	1957)	and	washed	in	running	water.	Later,	oysters	were	decalci-fied	by	immersion	in	a	solution	of	10%	EDTA	for	approximately	12	h	(Howard	et al.,	2004)	and	washed	in	running	water	to	eliminating	excess	of	EDTA.	Thereafter,	oysters	were	placed	in	histocassetes	for	histological	process	according	to	Shaw	and	Battle	(1957)	and	they	were	checked	for	 tissue	alterations	and	general	condition.	The	size	of	oysters	 from	different	subsamples	and	 reproductive	conditions	was	compared	for	possible	significant	differences	us-ing	the	t-test	(Zar,	1974).The	 shell	 height	 of	 the	 oysters	 from	 the	 first	 subsample	ranged	 from	 2	 to	 4.1	 mm,	 the	 mean	 value	 was	 2.99	 mm	 (±	 0.51	SD).	In	Figure	1A	is	showing	a	live	oyster	of	about	3	mm	of	shell	height.	In	64%	of	individuals	from	this	subsample,	we	do	not	de-tect	gonad	development	and	 this	sub-group	was	considered	as	undifferentiated	(Fig.	1B);	however,	36%	of	these	oysters	showed	development	 of	 reproductive	 follicles	 with	 sperm	 (Fig.	 1C).	 The	shell	height	of	this	sub-group	varied	from	2	to	4	mm	with	a	mean	value	of	2.95	mm	(±	0.52	SD).	There	was	no	significant	difference	between	the	shell	heights	of	both	subgroups	(t-test	p	>	0.05).	The	shell	height	of	oysters	from	the	second	subsample	varied	from	2	to	4.4	mm,	 the	mean	value	was	3.19	mm	(±	0.58	SD),	62%	of	 the	oysters	 showed	 gonad	 development,	 from	 which	 80.55%	 were	males,	13.89%	females	(Fig.	1D)	and	5.56%	hermaphrodites	(Figs.	1E,	1F).Figure	1A-F.	A.	Live	juvenile	Kumamoto	oyster.	B.	Undifferentiated	oyster	of	2.5	mm	of	shell	height.	There	is	no	follicular	develop-ment	in	the	connective	tissue	(Ct)	surrounding	the	digestive	gland.	C.	Male	oyster	of	2.7	mm	of	shell	height	with	sperm	(S)	sur-rounding	the	digestive	gland.	D.	Female	oyster	of	3.5	mm	of	shell	height	with	ovocytes	(Ov).	E.	Hermaphrodite	oyster	of	3.00	mm	of	shell	height.	F.	Detail	of	hermaphrodite	(encircled),	showing	ovogonies	(Og)	attached	to	the	germinal	wall	of	the	follicle	and	spermatids	and	spermatozoids	(Sp)	in	the	central	area	of	the	follicle.Notas	 183Vol.	22	No.	2	•	2012There	was	no	significant	difference	between	the	shell	height	of	both	subsamples	(t-test,	p	>	0.05);	however,	there	was	a	trend	of	larger	oysters	in	the	second	subsample.Results	indicate	that	gametogenesis	of	cultured	Kumamoto	oyster	starts	in	very	young	and	small	size.	This	finding	adds	an-other	 biological	 difference	 between	 C. gigas	 and	 C. sikamea,	the	later	starts	gamete	production	at	about	3	mm	of	shell	height	versus	16	mm	in	C. gigas.	 It	 is	 important	to	carry	out	studies	on	natural	populations	of	C. sikamea	for	determining	if	this	early	de-velopment	of	gametes	takes	place	in	nature	or	if	it	was	influenced	by	the	hatchery	operation,	possible	due	by	use	of	chemicals	dur-ing	culture	(Cooper	&	Wintermyer,	2009).	Presence	of	gametes	in	these	oysters	do	not	means	that	they	are	physiologically	viable.	Moreover,	 there	 was	 not	 observed	 empty	 follicles	 or	 the	 pres-ence	of	hemocytes	which	could	indicate	spawning	or	reabsorp-tion	process.	Specific	studies	are	needed	for	determining	if	these	small	oysters	may	reproduce	successfully	and	if	so,	how	this	early	reproduction	accounts	for	oyster	population	dynamics	in	nature,	and	 how	 this	 information	 may	 impact	 management	 practices	 in	hatchery	oyster	production.The	fact	that	the	percentage	of	oysters	showing	gametogen-esis	was	greater	in	oysters	from	the	second	sample	could	be	re-lated	to	their	slightly	larger	size;	in	this	sense,	Quayle	(1988)	noted	that	 in	C. gigas	sexual	maturity	appears	to	be	a	function	of	size	rather	than	age.	This	information	constitutes	a	new	record	in	the	size	of	gametogenesis	in	oysters	of	aquaculture	importance,	plac-ing	the	Kumamoto	oyster	(Crassostrea sikamea)	as	the	most	pre-cocious	species	among	cultivable	oysters	in	the	world.	Precocity	in	bivalve	mollusks	has	been	documented	in	Mytilus galloprovin-cialis Lamarck,	1819	(Mikhailov	et al.,	1995;	Paz	et al.,	2001).ACKNOWLEDGEMENTSThe	authors	would	like	to	thank	Ing.	Daniel	Carreño	Montreal	from	Sea	Farmers,	S.	A.	de	C.	V.	for	providing	us	with	the	oysters	for	the	study	and	allowing	us	to	publish	the	data	obtained.REFERENCESAmemiyA, i.	 1928.	 Ecological	 studies	 of	 Japanese	 oysters,	 with	 special	reference	to	the	salinity	of	their	habitats.	Journal of the College of Agriculture	9:	333-382.BAnks, m. D., D. J. mcGolDrick, W. BorGeson & D. HeDGecock.	1993.	Discrim-ination	between	closely	related	Pacific	oyster	species	(Crassostrea)	via	 mitochondrial	 DNA	 sequences	 coding	 for	 large	 subunit	 rRNA. 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Early Gametogenesis of Kumamoto oyster (Crassostrea sikamea)

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Early Gametogenesis of Kumamoto oyster (Crassostrea sikamea)

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