APPENDIX 2
Outside Activities

Oil-spill investigation in the strait of Malacca.

At the request of the Director General of Fisheries, Jakarta, I have participated as technical adviser on a cruise to investigate the effects of the oil spill from the Showa Maru accident in the strait of Malacca, 23–30 January 1975. The report on this investigation is to be finalized by the Indonesian members of the commission, and until now the report has not been published.
Technical advice has been given to Mr. P.H. Sand, legal consultant on environmental law to the Ministry of Justice, 14–16 August 1975. See Mr. Sand's consultant report: FAO, WS/H2676.
External help has been obtained for the analysis of Brestan-60 (triphenyl tin acetate), which pesticide is frequently used in aquaculture, but cannot be analysed by gaschromatography. Dr. Bozena Cosovic of the Ruder Boskovic Institute, Section on Polarography, Zagreb, Yugoslavia, was so kind to analyse 4 tambak sediments for us on tin residue. The results are:

Tambak	Date of Brestan-60 spraying	Tin ppm/dry	Brestan-60 equivalent ppm/dry
K-1	June Blank	-	-
E-4	June 1971	2.5	8.7
F-5	June 1974	3.5	12.1
F-2	June 1975	6.0	20.6

As long as the forms in which the tin residues occur (ionic or triphenyl form) are unknown, one has to regard the residue in the toxic triphenyl form, since this tin is arriving from Brestan-60 spraying. Ionic tin is considered as a non-toxic metal at least at such concentrations.

These initial results are not in favour for repeated applications of Brestan-60 in aquaculture, since the residues will accumulate in the sediment/water interface, which is also the source of food for Chanos.

Dr. Cosovic is highly acknowledged for this collaboration.

From a number of sediment samples from the Jepara bay, pesticide analyses have been made by Dr. J. Duinker and Mr. T. Hillebrand from the Netherlands Institution of Sea Research, Texel, The Netherlands. The results became available after finalizing this report, and are presented here below.

	BHC			pp'DDE	op'TDE	pp'TDE	pp'DDT	Dieldrin	PCB
	alpha	gamma	bétha	pp'DDE	op'TDE	pp'TDE	pp'DDT	Dieldrin	PCB
(1)	2.7	3.4	1.3	5.8	4.9	13.7	18.0	0.7	-
(2)	0.5	1.5	0.2	1.5	3.0	9.2	2.7	0.2	-
(3)	0.4	0.6	0.1	0.4	0.7	1.4	0.4	0.1	-
(4)	0.2	0.2	-	0.5	0.5	1.9	0.1	-	-
(5)	0.5	0.8	0.2	0.3	2.3	7.8	1.2	-	-
(6)	0.6	1.0	-	-	0.9	3.4	0.7	-	-
(7)	1.5	0.1	-	-	2.0	5.8	0.4	-	-
(8)	0.07	0.3	-	-	-	-	-	-	-
(9)	1.6	3.5	-	-	6.4	7.0	0.3	-	-
Results in ng/g dry				Samples taken 15/4/75			(ng/g-ppb)

The numbers 1 to 9 of the samples represent the sampling stations starting from a small river (1) and its mouth (2,3) further out in the estuary to 2 miles offshore (4–9).

The results are comparable with ours reported in Table VII-B concerning the order of magnitude of the concentrations, as well as on the spectrum of pesticides. PCB's were equally not detected. The results of Messrs. Duinker and Hillebrand show that the river is probably the source of the pesticides in the estuary.

Both Mr. Duinker and Mr. Hillebrand are thanked for their collaboration, which give support for the reliability of our measurements on the level of pesticide contamination in Indonesian littoral sediments.

A: Water

TABLE 1
Methods used for the analysis of pesticides in water, sediment and biological samples
Organo-phosphorus pesticides Diazinon and Leptophos
Organic Solvent		Water Quantity	Vial	Clean up	Injection μl
Kind	Quantity
Benzene	1 ml	10 ml	15 ml tube	-	10
Benzene or hexane	5 ml	200 ml	250 ml separatory funnel	-	10
Organo-chlorine pesticides
Organic Solvent		Water Quantity	Vial	Clean up	Injection μl
Kind	Quantity
Hexane	20 ml^a	200 ml	250 ml separatory funnel	Florisil Subsequent H₂SO₄	10

^a For low response, evaporated to 2 ml

B: Sediments

Organic Solvent		Sediment		Vial	Clean up	Injection μl
Kind	Quantity	Kind	Quantity	Vial	Clean up	Injection μl
Hexane	20 ml	freeze-dried	5 g	screwpot	Hg subsequently H₂SO₄	10
Hexane	200 ml	freeze-dried	40 g	Soxhlet	ibid	10
Acetonitril	200 ml	wet+Na₂SO₄	20+20 g	ibid	ibid, after extraction + 20 ml hexane + 100 ml H₂O	10 (hex)

C: Organisms

Organic Solvent		Sediment		Vial	Clean up	Injection μl
Kind	Quantity	Kind	Quantity	Vial	Clean up	Injection μl
For mainly organo-chlorine compounds:
Hexane	20 ml	fresh flesh liver stomach gonad total	2 g	screwpot	Florisil subsequently H₂SO₄	10
Hexane	20 ml	fresh flesh liver stomach gonad total	2 g	screwpot	Florisil subsequently H₂SO₄	10
Hexane	200 ml	freeze-dried	30 g	Soxhlet	ibid + eventually evaporation	10
Acetonitril	200 ml	Fresh+Na₂SO₄(1:1) total or different organs	30 g	ibid	ibid after extraction + 20 ml hexane + 100 ml H₂O	10 (hexane)
Acetonitril	20 ml	fresh + 1 h at 80°	2 g	screwpot	+ 10 ml hexane + 20 ml H₂O Florisil + subsequently H₂SO₄	10
For organo-phosphorus compounds:
Benzene	20 ml	fresh	2 g	screwpot	no or Florisil	10

TABLE 2

Comparison of various methods used. Symbols: I: fresh material directly treated with hexane; II: fresh material treated with acetonitrile and extracted with hexane; III: fresh material plus H₂O, cooked, extracted with hexane; IV: fresh material plus Na₂SO₄ (1:1 by weight), Soxhlet extraction by acetonitrile, re-extraction with hexane; V: freezedried, direct extraction with hexane; VI: freeze-dried, Soxhlet extraction with hexane. The applied clean-up procedures were either with H₂SO₄, or Florisil plus subsequently H₂SO₄, for sediment, the hexane extracts were first treated with mercury.

Turtle, Java Sea (Pulau Karimun Jawa)				(in ppb/dry)
	Method	BHC		DDE	TDE	DDT (p,p')	DIMU
	Method	Alpha	Gamma	DDE	TDE	DDT (p,p')	DIMU
Flesh	I	1.1	7.3	-	-	-
	IV	-	-	<1.9	<1.9	-
	V	4.3	12.3	-	-	-
	VI	23.8	-	-	-	-
Liver	V	4.3	-	3.2	-	3.7
Liver	VI	<20	<15	<13	<7.5	-
Mackerel (Canned from Japan)				(in ppb/wet)
(total)	I	63.7	-	56.3	133.7	-	44.8
	II	70.3	-	52.9	110.3	-	69.8
	V	33.0	-	38.5	74.7	0.2	2.3
	VI	64.6	-	44.9	78.5	5.9	30.9
Kuro (Polynemus sp.) marine fish, Jepara				(in ppb/dry)
(total)	I	2.7	3.4	370.5	122.8	150.3
	II	-	-	567.7	276.7	297.1
	IV	4.9	6.5	735.0	322.4	244.1
	V	27.4	4.0	162.3	86.4	52.2
	VI	48.2	-	400.5	291.6	149.0
Bandeng (Chanos chanos) marine, Jepara (about 1 m length)						(in ppb/wet)
Gonad	I	0.7	-	0.7	0.4	0.1
Gonad	IV	-	-	<5	-	-
Liver	I	10.3	-	1.7	1.8	-
	II	2.4	-	0.5	0.1	-
	IV	11.3	-	<20	2.4	7.8
	VI	-?-	-	1.6	1.9	1.5
Macrobrachium (Fresh-water shrimp) Jepara				(in ppb/dry)
	V	?	-	259.2	4.2	4.4
	VI	?	-	334.7	17.3	-
Pink shrimp (Metapenaeus monoceros) from West Irian					(in ppb/wet)
	I	1.4	-	0.5	-	-
	II	0.2	-	-	-	-
	III	0.2	-	-	-	-
	V	0.5	-	-	-	-
	Method	BHC		DDE	TDE	DDT (p,p')	Thiodan
	Method	Alpha	Gama	DDE	TDE	DDT (p,p')	Thiodan
Sediment from pond E₄ Jepara (was recently sprayed with BHC)						(in ppb/dry)
	I	2 364	-	0.5	1.5	0.3	0.4
	II	1 020	-	?	?	?	?
	IV	7 882	-	5.4	16.0	1.6	-
	V	7 710	-	?	0.8	0.3	7.0
	VI	11 347	-	?	17.6	3.2	4.6
Sundried Soxhl:		11 636	-	?	11.8	3.4	-
Bandeng (Chanos chanos) from pond E₄ Jepara (last spraying before sampling: 1½ month for BHC and 4 days for Diazinon) (in ppb/wet)
	I	1 678	-	19.1	14.0	8.5
	V	1 538	-	31.2	31.0	9.3
Na₂SO₄ -fresh -hex.		2 450	-	18.7	23.2	16.3
	IV	1 358	-	28.7	8.4	2.8
	VI	1 779	-	30.2	24.0	11.1	Diazinon
Freeze-dr.+benz.		FPD equipment					5 880

TABLE 3

Diazinon and Leptophos concentration as determined in the different specimens, detritus, water and absorbed to the aquaria (ppm/wet weight)

A. Diazinon

Time: (started 6/6/75) Concentrations Materials	hours			days						(days; all alive)				Total aquarium mg
Time: (started 6/6/75) Concentrations Materials	3	6	12	1	2	3	4	5		6	7	8	10	Total aquarium mg
added: 0.05 ppm									Still living fish removed into clean aquaria with ± 421 of water 20 ‰
Water	0.0075		0.013	0.0055	0.0017	0.0000	0.0007
Tilapia										0.0027	0.0026	0.000	0.000	0.000098
Chanos										0.0000	0.000	0.000	0.000⁺
Shrimp monoceros					0.000	Detritus		0.0073
Shrimp merguiensis		0.019		0.000
Chironomid						0.17
added: 0.5 ppm
Water	0.094			0.16	0.26	0.029	0.0000	0.0049
Tilapia										0.068	0.037	0.028	0.000
Chanos					0.000					0.0016	0.000	0.000	0.000
Shrimp monoceros		7.93				Detritus		0.39						0.00468
Shrimp merguiensis	0.72	0.070
Chironomid						0.36
added: 1.0 ppm
Water	0.24		0.36	0.27	0.046	0.00023	0.024
Tilapia										0.151	0.0146	0.0049	0.0017	0.277
Chanos					1.12	0.0355				0.349	0.000	0.0022	0.000
Shrimp monoceros		0.69				Detritus		2.98
Shrimp merguiensis	0.36	0.47
Chironomid					0.0184
added: 1.5 ppm
Water	0.45		0.51	0.27	0.104	0.0029
Tilapia				14.31	0.113
Chanos				0.817	0.0118			0.59					0.000	0.284
Shrimp monoceros	1.39					Detritus		0.025
Shrimp merguiensis	0.74	0.122
Chironomid					0.027
added: 2.0 ppm
Water	0.48			0.74	0.27	0.15	0.00082	0.064
Tilapia				13.31	0.164
Chanos				1.10	0.0034			6.10					0.000	0.811
Shrimp monoceros	1.91					Detritus		2.14
Shrimp merguiensis	0.39	0.157
Chironomid
added: 1.0 ppm
Contr. Water				0.22	0.10	0.056	0.0000	0.00091		0.0014	0.0009	0.0000	0.0009
Tilapia				6.01	4.15	4.93	?	3.09		1.01	0.380	0.0132	0.0007
						Detritus		0.037

B. Leptophos (Phosvel)

Time: Started 26/6/75 Concentrations Materials	hours			days					(days)						Total aquarium mg
	hours			days					(alive)				(dead)
	3	6	12	1	2	3	4	5	6	7	9	11	11	14
added: 0.05 ppm
Water	0.14			0.000	0.000	0.17?	0.000	0.000
Tilapia							0.21	1.06	0.68	1.08?
										0.28
Chanos							0.041	0.06	0.31	0.030	0.000	0.000			0.019
Shrimp monoceros		0.96	4.55
Detritus								0.000
Chironomid						41.69	32.64
added: 0.5 ppm
Water	0.76?			0.004	0.005	0.14?	0.000	0.002
Tilapia				2.71	11.00										0.606
Chanos							12.96	0.96	0.48	0.66	0.121	0.326	20.4	0.005
Shrimp monoceros		4.85	16.00
Detritus								41.60
Chironomid						73.33	58.02
added: 1.0 ppm
Water	0.28			0.054	0.015	0.052	0.003	0.005
Tilapia			6.31	5.94	14.67
Chanos							4.74	4.14	1.73	0.074	0.049	1.851	16.9-		1.363
Shrimp monoceros		2.27	8.10										20.0
Detritus								176.3
Chironomid						132.45	42.14
added: 1.5 ppm
Water	0.20			0.094	0.062	0.061	0.013	0.009
Tilapia				6.54	14.04
Chanos							16.52	5.57	11.31	0.92	0.94	0.521	17.2-	16.44	2.621
Shrimp monoceros	2.95												49.4
Detritus								566.9
Chironomid					16.55
added: 2.0 ppm
Water	0.46			0.330	0.162	0.190	0.022	0.386?
Tilapia			4.20	5.13	22.61
Chanos							17.82	4.96	4.82	6.83	2.63	5.972	82.6-		51.6
Shrimp monoceros	2.09												45.0
Detritus								1 801.8
Chironomid					66.00

TABLE 4

Budget and persistence of Diazinon and Leptophos at time t = 5 days for the aquaria toxicity tests. A: Diazinon, B: Leptophos

A: Diazinon

Specimens	Amount of Diazinon in μg
	Initial dosage
	0.05 ppm	0.5 ppm	1.0 ppm	1.5 ppm	2.0 ppm
M. monoceros	0.068	3.28	7.62	11.83	5.50
P. merguiensis	0.000	1.29	0.20	2.27	0.36
Chanos	0.000	0.01	0.81	3.98	9.32
Tilapia	0.051	0.78	0.19	14.41	54.07
Chironomid	0.018	0.43	0.002	0.004	?
Detritus	0.019	1.58	5.31	0.09	5.56
Water	28	196	960	4 040	2 560
Adsorbed	0.10	4.68	277	284	811
Total	28.256	208.05	1 251.13	4 356.58	3 445.81
Added (40 litre)	2 000	20 000	40 000	60 000	80 000
% Persistence	1.4	1.0	3.1	7.3	4.3

B: Leptophos

Specimens	Amount of Leptophos in μg
	Initial dosage
	0.05 ppm	0.5 ppm	1.0 ppm	1.5 ppm	2.0 ppm
P. merguiensis	1.40	4.50	0.86	2.46	1.16
Chanos	0.20	49.4	159.7	200.5	221
Tilapia	124.4	311.0	716.8	771.0	650
Chironomid	1.34	2.54	5.45	0.63	2.40
Detritus	0.00	21.3	321.9	859.3	1 164
Water	0.00	120	300	540	1 320
Adsorbed	19.0	606	1 363.0	2 621	51 600
Total	146.34	1 114.74	2 867.71	4 994.89	54 958.56
Added (60 litre)	3 000	30 000	60 000	90 000	120 000
% Persistence	4.9	3.7	4.8	5.5	45.8

APPENDIX 2 Outside Activities

APPENDIX 2
Outside Activities