ANNEXES

ANNEX 1 - Broodstock section: present consistency of female seabream and seabass stocks
(February 1995)

SEABREAM

biomas	number	hormonal treatments (per kg b.w.)		outdoor	recommended
(kg)		1993–4	1994–5	tank	for next season

20	16	none	2×LH-RHa 5μg	21	yes

40	34	none	1×LH-RHa 5μg and	21	yes
			1×hCG 500 I.U.

21	15	2×hCG 1000 I.U.	2×hCG 500 I.U.	15	no

23	16	2×hCG 1000 I.U.	1×hCG 500 I.U.	15	no

29	23	2×hCG 1000 I.U.	1×hCG 500 I.U.	16	no



SEABASS

biomass	number	hormonal treatments (per kg b.w.)		outdoor	recommended
(kg)		1993–4	1994–5	tank	for next season

7	5	none	1×hCG 500 I.U.	19	yes

8	4	none	1×hCG 500 I.U.	19	yes

13	6	none	1×hCG 500 I.U.	19	yes

83	39	1×hCG 1000 I.U.	none	19	yes

25	7	1×hCG 1000 I.U.	1×hCG 500 I.U.	14	no

50	?	1×hCG 1000 I.U.	none	17	yes

ANNEX 2 - Broodstock section: egg production, December 1994

Species: seabream											(egg weights expressed as grams)

day	tank 1		tank 2		tank 3		tank 4		tank 5		tank 6		tank 7		tank 8		tank 9
	V	NV	V	NV	V	NV	V	NV	V	NV	V	NV	V	NV	V	NV	V	NV


24	14	28	20	60	12	26	2	10	0	0	0	28	2	2	0	0	18	18
25	20	102	120	178	25	106	20	140	50	120	10	66	6	70	20	96	40	72
26	20	68	66	54	0	48	20	52	26	34	14	58	10	30	20	46	14	40
27	35	66	15	34	20	36	0	14	8	26	12	36	0	8	0	26	0	2
28	0	10	1	6	6	12	2	20	0	2	0	4	0	0	0	0	0	0
29		2						2
30
31


Total	89	276	222	332	63	228	44	238	84	182	36	192	18	110	40	168	72	132

											Total viable eggs:								668

											Total non viable eggs:								1858

ANNEX 3 - Broodstock section: egg production, January 1995

Species: seabream and seabass							(egg weights expressed as grams)

day	tank 1		tank 2		tank 3		tank 4		tank 5		tank 6		tank 7		tank 8		tank 9
	V	NV	V	NV	V	NV	V	NV	V	NV	V	NV	V	NV	V	NV	V	NV
1
2							2	2
3							64	28	40	40
4	12	4	54	4			32	50	68	32
5	224	88	198	102	206	54	0	62	20	40			22	22
6	336	138	298	132	382	118				22			10	0	0	16	0	4
7	174	122	186	94	226	120					bass		18	8	146	130	14	10
8	154	100	102	68	52	124					860	444	124	30	192	98	12	10
9	122	54	6	16	0	28					14	190	112	50	138	74	4	20
10	56	14	0	14	0	2							6	2	54	42
11	18	28	0	2	0	1					830	80	6	34	10	24	0	10
12	12	22	0	2	0	2	394	82	114	80	26	24	12	24	10	18	178	56
13	50	54	0	0	0	0	454	122	280	92							400	91
14	114	76	0	0	0	4	400	132	258	66							176	158
15	202	136	82	66	400	176	278	176	176	82							214	118
16	146	56	82	54	232	164	68	30	68	30							52	58
17	82	104	84	106	74	136	56	74	12	20							38	28
18	112	104	38	80	18	26	40	34	0	0							2	18
19	0	14	68	60	16	14	0	6	0	0							0	6
20
21
22
23
24
25
26
27											46	30	22	26	0	0	36	12
28											258	126	286	84	0	32	270	122
29											460	148	622	176	162	74	474	98
30											504	134	642	206	252	112	400	122
31											320	138	408	140	172	98	182	84
Totals:
	1814	1114	1198	800	1606	969	1788	798	1036	504	1588	576	2290	802	1136	718	2452	1025

													Total viable eggs seabream:					14908
													Total non viable eggs seabream:					7306
													Total viable eggs seabass:					1730
													Total non viable eggs seabass:					738

ANNEX 4 - Broodstock section: egg production, February 1995

Species: seabream and seabass										(egg weights expressed as grams)

day	tank 1		tank 2		tank 3		tank 4		tank 5		tank 6		tank 7		tank 8		tank 9
	V	NV	V	NV	V	NV	V	NV	V	NV	V	NV	V	NV	V	NV	V	NV
1											212	92	316	72	120	48	44	10
2											84	34	194	52	52	32	32	34
3
4
5
6
7											Seabass
8											1850	1020	1210	1548	3098	794	1458	1680
9											60	234	0	98	90	192	0	164



Total sebream eggs:											296	126	510	124	172	80	76	44
Total seabass eggs:											1910	1254	1210	1646	3188	986	1458	1844

													Total viable eggs seabream:					1054
													Total non viable eggs seabream:					374
													Total viable eggs seabass:					7766
													Total non viable eggs seabass:					5730

TOTAL EGG PRODUCTION:
			December			January			February			TOTALS
			V	NV		V	NV		V	NV		V	NV

Seabream			668	1858		14908	7306		1054	374		16630	9538		26168

Seabass			0	0		1730	738		7766	5730		9496	6468		15964

ANNEX 5 - Broodstock section: hormonal treatments, egg production and quality
(biomass and eggs expressed as kg; LH-RHa as μg/kg b.w.; hCG as I.U./kg b.w.)

batch	species	tank	treatment	date	female	total eggs	% biomass	% viable eggs	notes
					biomass
1	bream	1	LH-RHa/3.9	19-12-94	6.7	0.4	6.0	24.4
	"	"	LH-RHa/2.0	28-12-94	"	0.0	0.0	0.0	2nd treatment
2	"	2	LH-RHa/3.5	19-12-94	7.3	0.6	7.6	40.1
	"	"	LH-RHa/1.0	28-12-94	"	0.0	0.0	0.0	2nd treatment
3	"	3	LH-RHa/3.7	19-12-94	6.2	0.3	4.7	21.6
	"	"	LH-RHa/3.9	28-12-94	"	0.0	0.0	0.0	2nd treatment
4	"	4	LH-RHa/3.9	19-12-94	6.7	0.3	4.5	15.6
	"	"	hCG/500	28-12-94	"	0.2	3.7	40.8	2nd treatment
5	"	5	LH-RHa/3.9	19-12-94	6.9	0.3	4.3	31.6
	"	"	hCG/500	28-12-94	"	0.3	3.9	48.9	2nd treatment
6	"	6	LH-RHa/3.7	19-12-94	7.3	0.2	2.7	15.8	(*)
7	"	7	LH-RHa/3.9	19-12-94	7.0	0.1	1.4	14.1
	"	"	hCG/500	3-1-95	7.0	0.5	6.9	64.6	2nd treatment
8	"	8	LH-RHa/4.5	19-12-94	5.0	0.2	4.0	19.2
	"	"	hCG/500	3-1-95	6.6	1.0	14.4	57.8	2nd treatment
9	"	9	LH-RHa/4.4	19-12-94	6.9	0.2	2.9	35.3
	"	"	thermic shock	30-12-94	"	0.0	0.0	0.0	2nd treatment
	"	"	hCG/500	3-1-95	"	0.0	0.0	0.0	3rd treatment
10	"	1	hCG/500	2-1-95	7.3	1.7	23.3	66.4
	"	"	hCG/500	11-1-95	"	1.3	17.1	56.5	2nd treatment
11	"	2	hCG/500	2-1-95	6.6	1.3	19.4	66.0
	"	"	hCG/500	11-1-95	"	0.7	10.9	49.2	2nd treatment
12	"	3	hCG/500	2-1-95	7.2	1.3	18.3	65.9
	"	"	hCG/500	11-1-95	"	1.3	17.5	58.7	2nd treatment
13	bass	6	hCG/500	6-1-95	7.0	1.5	21.4	58.0
14	bream	4	hCG/500	9-1-95	7.9	2.3	29.6	72.0
15	"	5	hCG/500	9-1-95	7.5	1.3	17.0	71.0
16	"	9	hCG/500	9-1-95	7.6	1.6	21.0	66.1
17	bass	6	hCG/500	9-1-95	7.8	1.0	12.2	89.1
18	bream	6	hCG/500	26-1-95	8.4	2.6	31.0	72.9
19	"	7	hCG/500	26-1-95	6.7	3.2	48.4	76.7
20	"	8	hCG/500	26-1-95	7.6	1.2	15.2	65.7
21	"	9	hCG/500	26-1-95	6.7	1.9	28.7	74.9
22	bass	6	hCG/500	6-2-95	8.8	3.2	36.4	60.4
23	"	7	hCG/500	6-2-95	9.4	2.9	30.9	42.4
24	"	8	hCG/500	6-2-95	11.7	4.2	24.8	76.4
25	"	9	hCG/500	6-2-95	8.3	3.3	39.8	44.2

			Total seabream eggs		133.4	26.2	19.7	63.6
			Total seabass eggs		53.0	16.0	30.2	59.5

(*) Transferred in tanks 7, 8 and 9 for a 2nd treatment

ANNEX 6 - Broodstock section: fertility rates and eggs quality in seabream and seabass.
(LH-RHa as μg/kg b.w.; hCG as I.U./kg b.w.)

batches	treatment	date	eggs expressed as		notes
			% biomass	% viable eggs

SEABREAM

1 to 9	LH-RHa/3.9	19-12-94	4.2	24.2	1st treatment

1 to 3	LH-RHa/2.3	28-12-94	0.0	0.0	2nd treatment

4 & 5	hCG/500	28-12-94	3.8	44.9	2nd treatment

9	thermic shock	30-12-94	0.0	0.0	2nd treatment

7 & 8	hCG/500	3-1-95	10.7	61.2	2nd treatment

10 to 12	hCG/500	2-1-95	20.3	66.1	1st treatment

10 to 12	hCG/500	11-1-95	15.2	54.8	2nd treatment

14 to 16	hCG/500	9-1-95	22.5	69.7	1st treatment

18 to 21	hCG/500	26-1-95	30.8	72.6	1st treatment


SEABASS

13	hCG/500	6-1-95	21.4	58.0

17	hCG/500	9-1-95	12.2	89.1

22 to 25	hCG/500	6-2-95	33.0	55.9

ANNEX 7 - Broodstock section: compared fertility rates in 1993–4 and 1994–5
(biomass and eggs expressed as kg; hCG as I.U./kg b.w.)

season	species	treatment	female	total eggs	% biomass	% viable eggs
			biomass


1993–4	bream	1 × hCG/1000	66.1	13.9	21.0	74.8
		and 1 × hCG/500

1994–5	bream	2 × hCG/500	21.1	7.6	36.0	60.5
"	"	1 × hCG/500	23.0	5.2	22.6	69.7
"	"	1 × hCG/500	29.4	8.9	30.3	72.6


1993–4	bass	1 × hCG/1000	53.6	19.0	35.4	50.5

1994–5	bass	1 × hCG/500	53.0	16.0	30.2	59.5

ANNEX 8 - Broodstock section: average egg size in seabream and seabass.

	SEABREAM		SEABASS
	1994–5	1993–4	1994–5	1993–4	1995^o


Æ (mm)	0,96	0,92	1	1,1	1,08


weight (mg)	0,754 (Dec. 94*)	0,613	0,815 (Jan. 95)	0,908	1,218 (Jan. 95)
	0,702 (Jan. 95)		0,940 (Feb. 95)

* - LH-RHa induction

^o - Natural spawned eggs from a private farm

ANNEX 9 - Live food section: algal production results

		1994–5	1993–4	difference

Production (cu.m)		144,4	210	34,1% less




Algal growth in 350-I bags (million of cells/ml):

age	Nannochlor.	Isochrysis
(days)
0
1
2
3
4
5
6
7
8

Monthly algal production:

		0.5–1 flask	2-1 flask	6-1 balloon	65-1 bags	350-1 bags	production
							(liters)

November 1994		84	124	116	82	24	15000

December 1994		46	192	184	240	128	24500

January 1995		130	210	204	245	127	63000

February 1995		69	119	107	162	85	41900


							144400

ANNEX 10 - Live food section: daily controls of the section head

MORNING

general control of the unit:
watchman job during the night
air temperature (check YORK cooling system if something wrong)
oven
night oxygen (values in the tanks and bottles consume)
night carbon dioxide (bottles consume)
staff
plan of rotifer, algal and artemia production
choise of algae and rotifers for the larval feeding of the day after
supervision of algal replicates (small vessels and mass culture)
supervision of rotifer replicates (small vessels and mass culture)
supervision of artemia incubation, harvest and enrichment
consummable consumption:
oxygen bottles
carbon dioxide bottles
yeast
hypochlorite
hydochloric acid
chemicals for the algal culture
plastic bags

Inform the hatchery manager of any need in due advance, always keeping an amount of consummables suitable for at least 3–4 working days.

AFTERNOON

general control of the unit
quality control of rotifers and artemia for the larval unit
quality control of new rotifer tanks
quality control of the algal cultures to be inoculated the day after
instructions for the nightwatchman
plan the work of the day after

ANNEX 11 - Live food section: rotifer production results

Total production (billion):
		1994–5	1993–4	difference
				(%)

for larvae		55,7	33,3	67,3
for tank inoculum		105,2	88,7	18,6
discharged		42,3	151,2	72,1 less
	Total	203,2	273,2	25,6 less


Culture growth in 3000-1 fiberglass tanks:

	1994–5		1993–4
age	rotifers	samples	rotifers	samples	difference
(days)	(n/ml)	(n)	(n/ml)	(n)	(%)

0	215	68	181	54	18.8
1	254	74	230	54	10.4
2	341	70	303	54	12.5
3	462	68	483	54	4.3 less
4	476	68	570	54	16.5 less
5	404	62	602	34	32.9 less
6	569	15	402	4	41.5


Number of small vessel cultures:

		0.5 1 flask	2 1 flask	6 1 balloon	65 1 bags	350 1 bags

November 1994		41	48	29	14	4

December 1994		43	115	102	94	72

January 1995		39	125	121	102	58

February 1995		39	54	45	48	19

	total	162	342	297	258	153

ANNEX 12 - Live food section: artemia production results

Hatching results:
	average number/g cyst		production*
	1994–5	1993–4	(million)

nauplii AF	262000	356000	1364
nauplii BE	295000	318000	3745
nauplii EPAC	192000	_	5094
metanauplii	171000^o	n.a.	6816

* - Up to the date of 25/02/95

^° Calculated on an average survival rate of 89,1% of nauplii EPAC after 24-hour enrichment

ANNEX 13 - Live food section: artemia daily programme

Date

CYSTS TO BE INCUBATED :

* cyst AF : g into tank filled with liters

* cyst BE : g into tank filled with liters

* cyst EG : g into tank filled with liters

NAUPLII TO BE HARVESTED :

* AF nauplii from tank

* BE nauplii from tank

* EG nauplii from tank

METANAUPLII TO BE HARVESTED:

from tank at

NAUPLII TO BE ENRICHED (SUPERSELCO added at 10.30 and 22.30):

* Put million nauplii in tank filled with l; add g S/SELCO

ANNEX 14 - Live food section: consumption of expendable material in 1994–5 and foreseen needs for 1995–6

items	starting	total	residual	to be ordered	notes
	stock	consumption	stock	for 1995–6

NaNO₃ (kg)	75	46	29	40
KH₂PO₄ (kg)	29	3,1	25,9	_
NH₄CI (kg)	11	2,1	8,9	_
ZnSO₄×H₂O (g)	0,9	0,1	0,8	_
CuSO₄×5H₂O (g)	1,1	0,1	1	_
MnSO₄×H₂O (g)	0,9	0,1	0,8	_
CoSO₄×7H₂O (g)	0,5	0,1	0,4	_
FeCI₃×6H₂O (g)	0,8	0,1	0,7	_
Na₂MoO₄×2H₂O (g)	1	0,1	0,9	_
EDTA×2H₂O (g)	3,7	0,5	3,2	_
Vit.B12 (g)	8	1,8	6,2	_
Vit. B1 (g)	1023	170	853	_
Vit. H (g)	3	1,7	1,3	2
Selco (kg)	27	_	27	_
Super Selco (kg)	34	16,5	17,5	50	(^o)
Protein Selco (kg)	48	29,2	18,8	20	(^o)
Artemia AF	10,5	6,9	3,6	15	(^o)
Artemia BE	15,4	13,5	1,9	25	(^o)
Artemia EPAC	140	48,5	91,5	150	(^o)
Baker's yeast	_	724	_	800

(^o) - Figures updated to 25/02/95: final consumption will be higher.