BALTIC SEA ENVIRONMENT PROCEEDINGS

No. 27 D

GUIDELINES FOR THE BALTIC MONITORING PROGRAMME FOR THE THIRD STAGE

PART D. BIOLOGICAL DETERMINANDS

..·

BALTIC MARINE ENVIRONMENT PROTECTION COMMISS ION - HELSINKI COMMISS IO -

' .I BALTIC SEA ENVIRONMENT PROCEEDINGS

No. 27 D

GUIDELINES FOR THE BALTIC MONITORING PROGRAMME FOR THE THIRD STAGE

PART D. BIOLOGICAL DETERMINANDS

BALTIC MARINE ENVIRONMENT PROTECTION COMMISSION - HELSINKI COMMISSION - December 1988 PREFACE

The Guidelines for the Third Stage of the Baltic Monitoring Programme (BMP) are based on the Guidelines for the Second Stage of the BMP, published by the Commission as Baltic Sea Environment Proceedings No. 12 (BSEP No.l2). They have been revised by an expert group nominated by the Commission. The group was chaired by Dr. Gunni Aertebj erg and experts from all the Baltic Sea States participated in the work, with assistance from the International Council for Exploration of the Sea (ICES) and experts of the Baltic Marine Biologists (BMB).

·The nint.h meeting of the Helsinki Commission ( 15-19 February 1988) accepted the Guidelines in general as HELCOM Recommendation 9/7. The Commission recommends that the Governments of the Contracting Parties to the Helsinki Convention should apply the Guidelines for the Third Stage of the BMP, i.e. from 198 9 to 1993, and also, whenever possible, to follow the Guidelines in the monitoring of the internal waters as well. The data is to be submitted to the data bases of the Commission, as specified in the Guidelines.

The Guidelines for the Third Stage of the BMP are published in the BSEP series as four separate volumes (27 A, 27 B, 27 C, 27 D) and also as one combined volume of loose sheets.

The contents of the Guidelines for the Third Stage of the BMP is as follows: BSEP 27 A; Part A; Introductory Chapters 27 B; Part B; Physical and Chemical Determinands in Sea Water For bibliographic purposes this document should be dted as: Baltic Marine Environment Protection CommiSSIOn 27 C; Part C; Harmful Substances in Biota and Sediments - Helsinki Commission - 27 D; Part D· 1988 ' Biological Determinands Guidelines for the Baltic Monitoring Programme for the Third Stage; Part D. Biological Determinands Baltic Sea Environment Proceedings No. 27 D Volumes B, C and D are intended to be used together with Part A ' Copyright by the Baltic Mari~e Environr:r-ent Protection which contains general information on e.g. station networks, Commission - Helsmki Commission sampling requirements and data submission. ISSN 0357-2994

Helsinki 1988- Government Printing Centre

1 483184A ''"''<.f::;;z;-;;s"'Y}i!JFJJ£!4!;ii'W.'*. ------~

Any corrections or proposals for improvements concerning the GUIDELINES FOR THE BALTIC MONITORING PROGRAMME FOR THE THIRD STAGE content of these Guidelines are welcomed, and to be addressed to:

C 0 N T E N T S Baltic Marine Environment Protection Commission of the Baltic Sea Environment Proceedings No. 27 A, B, C, D - Helsinki Commission - Mannerheimintie 12 A A. INTRODUCTORY CHAPTERS SF - 00100 Helsinki (Baltic Sea Environment Proceedings No. 27 A) Finland Table of contents of Part A Tel.: 90 - 602 366 Tlx.: 125105 hlcom sf 1. Introduction 2. Station grid and maps Tfx.: 90 - 644 577 3, Sampling frequency and timetable 4. Data 5. Conditions required for carrying out scientific Possible comments concerning the formats prepared by the ICES research in the fishing/economic zone (for programmes should be addressed to the ICES, accordingly, as indicated in the adopted by the Helsinki Commission) 6. Format for notification of proposed research cruises formats. 7. Research vessels operating in the Baltic Sea Area

B. PHYSICAL AND CHEMICAL DETERMINANDS IN SEA WATER (Baltic Sea Environment Proceedings No. 27 B)

Table of contents of Part B

B. I Basic Hydrographic and Hydrochemical Determinands

1. Temperature 2. Salinity 3. Density structure 4. Oxygen 5. Hydrogen sulphide 6. pH 7. Alkalinity 8. Nutrients 9. Sampling and preservation of samples 10. References in Chapter B I 11. Reporting format for hydrographic and hydrochemical data

B. II Heavy Metals, Petroleum Hydrocarbons and Chlorinated Hydrocarbons in Sea Water

1. Heavy metals 2. Petroleum hydrocarbons (PHCs) 3. Chlorinated hydrocarbons (CHCs) 4. Sampling and preservation of samples 5. References in Chapter B II 6. Reporting format for contaminants in sea water 7. Exchange of data'on contaminants in sea water via magnetic tape c. HARMFUL SUBSTANCES IN BIOTA AND SEDIMENTS TABLE OF CONTENTS OF PART D (Baltic Sea Environment Proceedings No. 27 C)

Table of contents of Part C D. BIOLOGICAL DETERMINANDS Page

C. I Harmful Substances in Biota 1. Phytoplankton primary production 2 1. Harmful substances in selected species 2. Phytoplankton chlorophyll-~ and 16 2. Analytical procedures for mercury, cadmium and lead in phaeopigments biological material 3. Phytoplankton 23 3. Chlorinated hydrocarbons in biological material 4. Checklist of phytoplankton 61 4. References in Chapter C I 5. Rubin codes of phytoplankton 87 5. Reporting format for contaminants in marine biota 6. Phytoplankton identification sheets 87 6. Exchange of data on contaminants in fish and shellfish 7. Zooplankton 88 via magnetic tape 8. Macrozoobenthos 91 9. Micro-organisms 101 C. II Trend Monitoring of Contaminants in the Coastal Zone 10. Biological data reporting format 116 11. Forms to be used in relation to 154 1. Choice of monitoring organisms the biological data reporting format 2. Sampling procedures 12. Microbiological data reporting format 161 3. Subsampling and handling 4. References in Chapter c II Tables: c. III Harmful Substances in Sediments Tab.D.l. The factor, F, for calculating total 10 from carbonate alkalinity, temperature, 1. Contaminants in sediments co2 2. Reporting format for contaminants in sediments pH, and salinity Tab.D.2. Stereometrical formulas for common phyto­ 42 plankton taxa in the Baltic D. BIOLOGICAL DETERMINANDS (Baltic Sea Environment Proceedings No. 27 D) Figures: Table of contents of Part D Fig.D.l. Girdle view of a centric diatom with 52 1. Phytoplankton primary production diameter d, height h and plasma 2. Phytoplankton chlorophyll-a and phaeopigments thickness c 3. Phytoplankton - Fig.D.2. Flow chart of TTI-measurement technique; 108 4. Checklist of phytoplankton sample volumes, incubation times and 5. Rubin codes of phytoplankton liquid scintillation (LSC) procedure 6. Phytoplankton identification sheets may vary between sites 7. Zooplankton 8. Macrozoobenthos 9. Micro-organisms 10. Biological data reporting format 11. Forms to be used in relation to the biological data reporting format 12. Microbiological data reporting format -1-

D. BIOLOGICAL DETERMINANDS

Biological determinands to be monitored

Phytoplankton primary production

Phytoplankton chlorophyll-~ and phaeopigments *) Phytoplankton (species composition, number of counting units, biomass) Zooplankton (species composition, abundance and biomass of mesozooplankton, protozooplankton *)) Soft bottom macrozoobenthos (species composition, abundance, biomass) Micro-organisms *) (total number and biomass of bacteria, production of bacteria, number of colony­ forming bacteria)

New sampling methods: To improve the significance and reliability of pelagic sampling, the use of additional new sampling methods is recommended, such as automatic sampling and sediment trapping.

Remark: It is essential that sampling of macrozoobenthos is accompanied by some hydrographic measurements to provide information about the hydrographic situation. Therefore, as a minimum requirement, water should be sampled as close to the sea bottom as possible for determination of salinity, temperature and oxygen/H2s concentration. Preferably a complete hydrographlc series should be taken.

Sampling depths

The sampling depths are described under the sections for each determinand.

*) tentative determinands -2- -3-

1. Phytoplankton primary production Samplers

Introduction Non-transparent and non-toxic sampling devices must be used. In order to make it possible to calculate the daily 2 phytoplankton primary production per m water surface Experimental bottles by the data consultant the following measurements are obligatory and shall be reported: 25 cm 3 bottles made of high quality laboratory glass Temperature corrected potential production at the and with standard grinding and glass stoppers are sampling depths, measured in incubator recommended. Irradiance in the incubator used Vertical irradiance attenuation in the water at the The experimental bottles must be thoroughly cleaned sampling station, measured either by Secchi-disc or before every experiment in order to avoid bacterial an irradiance meter. film ox; adsorption of toxic substances to the inside of the bottles. The bottles must be cleaned with a 10 % a) Production measurements HCl-solution, then rinsed in tapwater and in distilled water. If possible the bottles should be dried at Sampling depths 170°C. Before use the bottles must be washed with water from the respective samples. Sampling depths are selected to give an adequate vertical production curve. The standard sampling depths All handling of samples before and after the incubation are l m, 2.5 m, 5 m, (7.5 m,) 10m, 15m and (20m) experiment must take place in dimmed light. (non-obligatory depths in brackets). The incubator experiment must be carried out as soon as In water bodies with a thermocline and/or a halocline, possible after sampling. a higher concentration of phytoplankton is often ob- served in the discontinuity layer than above or below From each sampling depth, two clear experimental this layer. If a pycnocline is found within the bottles are filled with water. Additionally, two dark euphotic zone and does not correspond to one of the experimental bottles are filled with water from 1 m and standard sampling depths, it is recommended to collect 15 m depths, respectively. an additional sample in the discontinuity layer. ,. determination of the irradiance production Sampling time celation (the PI-curve) seven clear experimental bottles are filled with water from 1 m depth or, with Water samples for production measurements should water from the integrated phytoplankton sample (see preferably be sampled between 8 a.m. and 4 p.m. Central Section D. 3. a)). If a pycnocline is found within the European Time. euphotic zone, additionally seven clear experimental bottles are filled with water from 15 m depth. .·:cwo);>;;~------~·

-4- -5-

The experimental bottles must not be filled totally, Incubator but space for the 14c-solution and a little air bubble should be left. The incubator used must have the following specifications: 14c-solution - thermostatically controllable 14 The c-ampoules for use in production studies can be - irradiance conditions ensuring photosynthetic 18 -2 -1 purchased from different manufacturers. These ampoules saturation: at least 250. 10 quanta m • s . -2 -1 must fulfill the following specifications: (400-700 nm) or 100 JOUles m • s (400-700 nm). (Philips TLD 18 W/33 meets these demands). - alkalinity 1.5 mM/dm' specific activity 4 - 20 !" Ci/cm' Measurements of irradiance in the incubator

14 Standardization of c -solution A calibrated irradiance meter (quanta meter, 400-700 nm) shall be used. Liquid scintillation counting shall be used as the basis for determination of the absolute activity. The irradiance meter is placed in the water-filled incubator facing the light source of the incubator and 14 . Concentration of c-solut1on at the same distance from the light source as the experimental bottles during experiments. The 14c-solution should be added to the experimental bottles in such concentrations that statistically suf­ The irradiances are measured in five different pos­ ficient estimations of the radioactivity fixed by itions: At the center of the bottle-wheel of the photosynthesis in the sample can be obtained. However, incubator and at the outermost positions of the it is also important not to disturb the co equilibrium experimental bottles on each side and above and below 2 14 in the water sample by adding too much NaH co3 the center of the bottle-wheel. solution. The measured irradiances are corrected for the Concentrations corresponding to a 1 em ' 14 C-so1 ut1on . immersion effect by multiplying by the immersion factor with a radioactivity of 1 - 4 ~Ci/cm' per 25 em' sample of the irradiance meter used. have been shown to be applicable for primary pr•)duction studies in the Baltic. Immersion factor: The ratio between the sensitivity of the irradiance meter in air and in water. Dark fixation of carbon The mean of the four outermost measurements are As the dark fixation of carbon is not directly related calculated, and the irradiance ·in the incubator is ' to photosynthetic production, it has to be reported expressed as the mean of the center measurement and the separately. calculated mean of the four outermost measurements. ''?)iJ.~%~'1.-""""------

-6- -7-

Incubation temperature Filters with even distribution of pore size, and good solubility with respect to scintillation liquids are The temperature in the incubator during the experiment preferred. Pore size should not exceed 0. 4 5 ~m. The has to be adjusted to the mean temperature of the filters should be wetted before the filtration starts. euphotic zone, or to the mean temperature found above 5 2 an eventual pycnocline if a pycnocline is present The suction pressure should not exceed 0. 3 10 Nm - within the euphotic zone. (0.3 atm.).

Incubation The whole filtration procedure should not exceed 0. 5 hour for the entire series of bottles. It is possible The experimental bottles are placed at the bottle-wheel to comply with this requirement by arranging a series of the incubator in such a manner that only clear ex­ of filtration units. In case it is impossible to filter perimental bottles face the irradiance source. the whole contents of a single sample, a subsample may be filtered. The subsample shall be of at least 15 em'. Five of the experimental bottles in each of the series If filtering a subsample, both the volume of the sub­ for determination of the PI-curves are covered with sample and the whole volume of the experimental bottle neutral filters of different transmissions, for example has to be measured. In the case of filtering the whole 5 %, 10 %, 15 %, 25 % and 50 %. The exact transmission volume of an experimental bottle, such measurements are should be known. Further, the production is determined not necessary. at 100 % incubator irradiance by the 6th bottle of the series and at 17 5 % incubator irradiance by the 7th The filters should not be washed but, whenever bottles bottle of the series. The 175 % incubator irradiance is and filtration funnels need to be rinsed, this should obtained by applying a tinfoil coating as reflector occur at the end of the filtration procedure, but behind the experimental bottles. before the last em' has passed through the filter.

Incubation period Preparation of filters

The incubation period is 120 minutes. In order to stop all biodegradation and thus losses of 14 radioactivity, and to remove possible c precipitates Filtration of samples extracellularly, immediately after filtration the wet membrane filters are placed in a desiccator and exposed The samples should be filtered immediately after the to vapours of fuming HCl for 3 minutes. production experiment is stopped, in order to avoid loss of 14c due to respiration. When scintillation counting is used, the filters can now be placed 'at the bottom of the empty scintillation vials. /,-:,;;.;y¢.,.""·------

-8- -9-

When Geiger counting is used, the filters must be dried TA = l. 26 + 0.031 s for 18 < s < 25 in a desiccator with freshly dried silicagel and with TA = 0.90 + 0.080 s for ll

to the amount and nature of filtered material Table D.l. The total co2 can then be expressed: (quenching and self-absorption problems). C02 = Carbonate alkalinity x F. The counting results should be given as disintegrations per minute (dpm). If more accurate co2 values are requested, which is recommended at higher salinities (the Kattegat and Belt Total concentration Sea) and also in coastal areas, where the relations co2 given by Buch (1) are less suitable, they can be For estimating the total carbon dioxide concentration, estimated according to Strickland & Parsons (11), the formulae given by Buch (1) and reproduced by Gargas Glowifiska et al. (4) or Mackereth et al. (9). (2) are recommended:

2 4S3184A -10- -11-

TABLE D.l. The factor, F, for calculating total co2 , b) Production calculations from carbonate alkalinity, temperature, pH, and salinity. Calculation of carbon uptake

The total carbon uptake, Pt during the time t, is calculated for all of the experimental bottles from the pH 7.0 7.1 7.2 7.3 7.4 7.5 7.6 7.7 7.8 7.9 8.0 8.1 8.2 8.3 8.4 following equation: s •/00 1 1.284 1.234 1.189 1.150 1.117 1.093 1.074 1.057 1.043 1.031 1.022 1.014 1.008 1.002 0.996 4 1.240 1.196 1.154 1.121 1.095 1.074 1.057 1.049 1,032 1,022 1.013 1.004 0.997 0.989 0,982 7 1.212 1.158 1.132 1.104 1.082 1.064 1.050 1.037 1.025 1.015 1.006 0.998 0.990 0,981 0.971 dpm(a).total co (c). 12(d).l.05(e).l.06(f).k ~ ~ 0 2 11 1.192 1.154 1.120 1.094 1.073 1.056 1.041 1.028 1.016 1.007 0.998 0.990 0.981 0.971 0.960 1 2 3 18 1.170 1.133 1.104 1,080 1.060 1.046 1.032 1.020 1.009 0.998 0.988 0.977 0.966 0,954 0.940 dpm(b) 25 1.152 1.107 1.087 1.067 1.048 1.034 1.020 1.010 0.998 0.987 0.976 0.964 0.952 0.938 0.923 1 1.271 1.223 1.175 1.136 1.104 1.084 1.066 1.050 1.040 1.030 1.020 1.011 1.004 0.998 0.992 4 1.220 1.180 1.140 1.109 1.084 1.065 1.050 1.036 1.024 1.016 1.008 1.000 0.992 0.984 0.978 7 1,192 1.153 1.120 1.093 1.074 1.056 1.042 1.030 1.020 1.010 1.000 0.992 0.984 0.974 0.964 where: 5 11 1.172 1.138 1.107 1.084 1.066 1.049 1.034 1.021 1.011 1.002 0.993 0.986 0.975 0.964 0.952 3 18 1.152 1.118 1.090 1.066 1.050 1.037 1.024 1.012 1.000 0.991 0.981 0.970 0.956 0.944 0.930 Pt =total carbon uptake, mg c m- h-l 25 1.132 1.102 1.076 1.056 1.040 1,028 1.016 1.004 0.992 0.982 0.970 0.958 0.944 0.928 0.908 1 1.244 1.190 1.152 1.121 1.098 1.078 1.062 1.048 1.036 1.026 1.016 1,008 1.000 0.993 0.988 4 1.204 1.160 1.126 1.100 1.080 1.063 1.048 1.034 1.024 1.015 1.006 0,996 0.988 0.981 0.973 7 1.174 1.138 1.110 1.086 1.068 1.052 1.038 1.026 1.016 1.007 0.998 0.988 0.980 0.970 0.960 (a) = filter dpm - background dpm = net dpm/filter 10 11 1.156 1.122 1.096 1.076 1.060 1.044 1.031 1.020 1.009 0.999 0.988 0.978 0.968 0,956 0.944 14 18 1.136 1.104 1.082 1.062 1.046 1.030 1.018 1.008 0.998 0.988 0.978 0.966 0.952 0.938 0.921 (b) = the activity of the added c-solution, dpm 25 1.118 0,915 1.092 1.071 1.052 1.034 1.020 1.008 0.997 0.986 0.974 0.962 0.950 0.933 0.896 (c) = concentration of total co in the experimental 1 1.229 '1.178 1.143 1.114 1.090 .1.070 1.056 1.041 1.030 1.020 1.010 1.002 0.996 0.989 0.984 2 4 1.188 1.144 1.115 1.091 1.074 1.056 1.041 1.030 1.020 1l. 010 ; 1.001 0.992 0.985 0.978 0.971 3 7 1.160 1.128 1.101 1.079 1.060 1.046 1.034 1.022 1.011 1.002 0.992 0.984 0.975 0.984 0.952 water, mM/dm 15 11 1.141 1.114 1.089 1.066 1.050 1.036 1.023 1.012 1.002 0.994 0,985 0.975 0.964 0.951 0.936 3 18 1.126 1.092 1.075 1.056 1.040 1.027 1.014 1.004 0.993 0.982 0.961 0.958 0.944 0.928 0.912 (d) = 12: the atomic weight of carbon, converts mM/dm 25 1.108 0,971 0.942 0.924 0.902 0.884 1.062 1.060 1.043 1.028 1.015 1.004 0.994 0.982 0.958 to mg/dm 3 1 1.210 1.166 1.130 1.102 1.080 1.084 1.050 1.040 1.028 1.018 1.010 1.001 0.992 0.984 0.978 4 1.170 1.130 1.106 1.084 1.068 1,054 1.040 1.029 1.018 1.007 0.996 0.986 0.976 0.966 0.959 14 7 1.144 1.114 1.090 1.072 1.056 1.040 1.028 1.018 1.008 0.998 0.988 0.978 0.968 0.956 0.944 (e) = a correction for the effect of c discrimi­ 20 11 1.130 1.100 1.080 1.062 1.045 1.031 1.020 1.010 1.000 0.990 0.980 0.968 0.956 0.943 0,930 14 18 1.112 1.086 1.066 1.050 1.034 1.021 1.009 0.998 0.986 0.974 0.962 0.950 0.936 0.920 0.901 nation, the uptake of the c-isotope is 5 % 25 0,976 0.896 0.872 1.098 1.064 1.054 1.040 1.024 1.011 1.000 0.988 0.962 0.948 0.932 0.916 slower than that of the 12c-isotope (f) = a correction for the respiration of organic matter produced during the experiment. This has empirically been found to represent 6 % at optimal photosynthesis. The production rate is thus corrected to represent the rate of gross production

3 k1 = a correction factor for subsampling, e.g. 15 cm were filtered from a sample of 27 cm 3 and 1 cm3 14 NaH co3 was added, then k 1 = 28/15 = 1.87 a time correction factor, e.g. when converting production per 2 hours to production per hour, then k ;, 0.5 2 a unit conversion factor, e.g. when converting mg -3 ; -3 3 C dm to mg C m , then k = 10 . 3 ~~,,______

-12- -13-

Subtraction of dark fixation c) Measurements of vertical irradiance attenuation

At stations with homogeneous water within the euphotic Introduction zone, the mean carbon uptake in the two dark exper- imental bottles is subtracted from the total carbon In order to make it possible to calculate the daily uptake in all the clear experimental bottles. At phytoplankton primary production per m' water surface stations with a pycnocline within the euphotic zone, by the data consultant, the following measurements are the dark-uptake at the 1 m depth is subtracted from all obligatory and shall be reported: the clear experimental bottles from the pycnocline and temperature-corrected potential production at the above, and the dark-uptake at 15 m depth is subtracted sampling depths, measured in incubator, from all the clear experimental bottles from below the irradiance in the incubator used, pycnocline. vertical irradiance attenuation in the water at the sampling station, measured either by Secchi disc or Calculation of potential production a qj.lanta meter.

The potential production at a sampling depth is ex- Secchi disc measurements -3 -1 pressed in mg C m h , and calculated as the mean of the production rates measured in the 2 clear experi­ In cases when irradiance meters are not available, a mental bottles from that depth and exposed to 100 % Secchi disc must be used to estimate the vertical incubator irradiance, with the dark fixation sub­ attenuation of irradiance in the water column. tracted. When a Secchi disc is used, the Secchi depth measured Temperature correction must be corrected for the wave height:

The potential production rates obtained must be cor­ Do = DH (1 + 0.4.H) m, where rected for eventual differences between the mean tern- Do = Corrected Secchi depth, m perature in the incubator during the experiment and the DH = Measured Secchi depth, m in situ temperatures at the sampling depths. The H = Wave height, m temperature correction factors are determined from the equation: The corrected Secchi depth, D 0 equals approximately the 10 % quanta depth.

Temp. correction factor= exp. (ln 2 x (t2 -t1 )) 10 The relative irradiance at a given depth, or the depth for a given relative irradiance, is determined by

where t 1 = the mean temperature in the incubator during graphical in~erpolation. On semi-logarithmic paper the experiment (irradiance: log scale, two decades; depth: linear t = the temperature at the sampling depth 2 scale), the curve becomes a straight line through 100 % irradiance at 0 m depth and 10 % irradiance at Do m depth. ,,,,,,,,~------

-14- -15-

Irradiance meter measurements The relative irradiances in the different water depths are calculated, setting Ed (z=o) = 100 %. If possible, instead of Secchi disc measurements, the vertical attenuation of irradiance in the water at the The relative irradiance at a given depth, or the depth sampling stations should be measured using a quanta for a given relative irradiance, is determined by meter (400-700 nm). graphical interpolation: The measured relative ir­ radiances are depicted (log scale) as a function of the The underwater detector is lowered from the side of the water depth (linear scale) on semi-logarithmic paper (2 ship facing the sun and so far from the side of the decades), and a smooth curve is drawn through the ship that the disturbance of the normal distribution of points. light is minimized. d) Data reporting The irradiance above the water surface, Ed (air), is

measured with the dry underwater detector. The Al~ data should be reported according to the details irradiance immediately below the water surface, Ed given in Sections D.lO and D.ll. (z=o), is then calculated as 93 % of Ed (air). (The mean reflection of irradiance by the water surface e)) References in Chapter D.l. approximates 7 %.)

1. Buch, K. 1945. Kolsyrejamvikten i Baltiska havet. The underwater detector is lowered into the water and Fennia 68(5). 1-208. the depths and irradiances are measured at the sampling 2. Gargas, E. (ed.) 1975. A Manual for phytoplankton depths of primary production. primary production studies in the Baltic, BMB Publ. 2. 1-88.

All underwater irradiance readings are corrected for 3. Gargas, E. and I. Hare 1976. User's manual for the immersion effect by multiplying the reading by the estimating the daily phytoplankton production measured in an incubator. Contr. Water Qual. Inst. immersion factor of the underwater detector used. 2. 1-75. Immersion factor: The ratio between the sensitivity of 4. Glowifiska, A., s. Ochocki, B. Popowska, H. Renk the detector in air and in water. and H. Torbicki 1975. Studia por6wnawcze nad meodami oznacznia weg1a nieorganiczego w badaniach produkoji pierwotnej (Comparative In immediate connection with every measurement with the studies on methods of inorganic carbon underwater detector, the irradiance in air is measured determination in research works on primary production). Stud. Mater. MIR 15. using a deck detector. From the deck detector measurements, the underwater measurements are corrected 5. Guidelines for the measurement of phytoplankton primary production. BMB Publ. 1, 2nd ed. 1984. for eventual changes in the irradiance above the water surface after the measurement in air with the under­ 6. Guidelines for the measurement of primary produc­ tion in the ICES area. C. M. 1981/L:46. water detector. ':.--,Z"J!:'f}%;o''______

-16- -17-

7. Hojerslev, N.K. 1979. Daylight measurements Non-transparent and non-toxic sampling devices must be appropriate for photosynthetic studies in natural sea water. J. Cons. int. Explor. Mer., used. 38. 131-146.

8. Incubator method for determining the carbon b) Storage of water samples f~similation by plankton algae using C-technique. Danish Standards Association. DS 293, 1983. 1-18. It is important that the water is filtered as soon as possible. If the filtration cannot be carried out im- 9. Mackereth, F. J. H. , J. Heron and J. F. Talling 1978. Water analysis. Freshwater Biological mediately, the sample must be stored cool and dark, Association. Sci. Publ. No. 36. preferably in a refrigerator, and no longer than 8 10. Photosynthetically active irradiance in water. hours. Danish Standards Association, DS 294, 1983. 1-10 (in Danish). c) Volume to be filtered 11. Strickland, J. D. H. and T. R. Parsons 1972. A practical handbook of seawater analysis, 2nd ed. Bull. Fish. Res. Bd. Can. 167. 1-310. In the,B~ltic Sea Area the chlorophyll a concentration varies from about 0.1 mg m- 3 during win~er and in the deep water to 15 mg m-3 or more in the surface water during the spring bloom. During summer the concen- 2. Phytoplankton chlorophyll-a and phaeopigments tration in the surface water is most often between 1 mg 3 -3 m- and 5 mg m

Measurements of phytoplankton chlorophyll-~ are -3 3 obligatory, while measurements of phaeopigments are At a concentration of 0.1 mg m and the use of 10 cm only tentative. extraction solvent and a 5 em cuvette, at least 12 dm 3 have to be filtered to get a spectrophotometric a) Sampling absorbance of at least 0.05.

3 Water for chlorophyll-~ analyses should be taken from When using 10 cm extraction solvent and a 1 em cuvette -3 the same samples as for primary production and at chlorophyll-a concentrations of 1 mg m and 10 -3 - 3 phytoplankton and, if possible, for chemical analyses. mg m at least 6 dm and 0.6 dm 3 water, respectively, have to be filtered to get absorbances of at least The standard sampling depths for chlorophyll-~ are the 0.05. same as for primary production: 1 m, 2. 5 m, 5 m, (7.5 m,) 10m, 15m and (20 m), non-obligatory depths d) Filtration being given in brackets. The samples shall be filtered in subdued light through Chlorophyll should also be analysed from the integrated Whatman GF/C filters. phytoplankton sample (see Section D.3.a)). Additional 2 chlorophyll sampling depths are recommended, e.g., one The suction pressure should not exceed 0. 5 • 1 o5 Nm - just above and one below the halocline. (0.5 atm). After filtration the filter shall be folded together. ":--,,,W,'1.'>-JM=------~

-18- -19-

The filtration time should not exceed 1/2 hour per h) Centrifugation sample. If this is not possible, a smaller water volume has to be used. If necessary, the extract volume is adjusted to 10 em' before centrifugation. The stoppered centrifuge tube is e) Drying of filters shaken vigorously to get a homogeneous distribution of the chlorophyll in the extraction solvent. As drying of filters has been shown in several experiments to increase the amount of extractable The sample is centrifuged for 10-20 minutes at about 2 chlorophyll pigments, it is recommended to dry the 10,000 m s- , in order to reduce the spectrophotometric folded filter wrapped in a piece of clean filter paper blank reading (750 nm) which should not exceed 0.005 in a room temperature airstream in darkness. for a 1 em cuvette.

f) Storage of filters i) Storage of extract

The filters ought to be analysed immediately after The measurements shall be made immediately after drying. centrifugation. If this is not possible the extract may be stored in a deep freezer (-20°C) for no more than 24 If storage is necessary, the filters shall be stored hours. deepfrozen (-20°C) in a desiccator with silica gel, for no longer than three months. j) Chlorophyll-a measurement procedure

g) Extraction The measurements of chlorophyll-~ may either be made by a spectrophotometer or a fluorometer, depending on the 96 % ethanol shall be used as extraction solvent. equipment available. The measurements of phaeopigments shall only be made by a fluorometer. All work with the chlorophyll extract shall be carried out in subdued or green light. 1) Spectrophotometric readings

Storage of the extract shall be in total darkness. A spectrophotometer with a bandwidth of 2 nm should be used. 96 % ethanol should be used as a reference. The folded filter is transferred to a graduated centrifuge tube, 10 em'. 96 % ethanol is added, and the --to-cell blanks should be measured at all wave­ tube is stoppered to avoid evaporation. Lengths used. The absorbence should be measured at 750 nm and 663-665 nm (at the peak). Extraction shall last 24 hours at room temperature in total darkness. The samples are shaken a few times For the measurement of phaeopigment, which is ' during the extraction time. non-obligatory, the extract shall be acidified with 1 M -20- -21-

HCl ( 0. 06 cm3 to 5 cm3 of extract) after all other A fluorometer with excitation setting of 425-430 nrn and readings have been taken. 0. 5-3 min. after acidifi­ emission setting of 663-665 nrn, or a filter fluorometer cation, the absorbence at 750 and 663-665 nm should be equipped with a blue lamp corresponding to GE F4T5B, measured. red-sensitive photomultiplier and primary filter corre­ sponding to Corning 5-60 and secondary filter corre­ Calculations sponding to Corning 2-64 should be used.

To calculate the chlorophyll-~ content using the The fluorescence shall be measured with the appropriate spectrophotometric technique, the following equation slit. should be used: For the measurement of phaeopigment the extract shall 3 be acidified with HCl (0.06 crn 3 to 5 crn 3 of = 10 .e. A(665 k) 1 M 83. v. 1 extract) after the first reading. 0. 5-3 minute after acidification, a new reading should be made.

3 cv = Chlorophyll-~ concentration, mg/rn Calculations e = volume of ethanol, ern' A( 665 k) = absorbence at 665 nrn (the peak) minus the absorbence at 750 nrn after correction by the Use the equation: cell-to-cell blanks -3 -1 Chl.-~ (rng.rn ) = R.f.s.e.V 1 = length of cuvette, ern v = water volume filtered, drn' R fluorescence reading 83 = absorption coefficient in 96% ethanol = f = calibration factor s correction The volume of chlorophyll sample, volume of ethanol and = slit e volume of ethanol (ern' ) the length of cell (cuvette) must be chosen to give ab­ = v volume of filtered water ( drn' ) sorbence values at 663-665 nrn of 0.05-0.8, i.e. the = optimum range of the spectrophotometer. The calibration factor is determined as follows:

2) Fluorornetric reading -1 -1 f = K.R .v.e The sample vol urne and the quantity of ethanol hco· ,; o be K = concentration of chlorophyll-~ (rng.rn-3 ) deter­ chosen so that the fluorescence reading is • · ~ optimum range of the equipment used (normally 50 · .c (). mined spectrophotometrically as described in Section D.2.k) and (1). crn 3 water and 5-10 ern' ethanol). ""'''''~'------

-22- -23-

When phaeopigment is to be calculated use the equation: 4. Lenz, J. & P. Fritsche 1980. The estimation of chlorophyll !:, in water samples: a comparative study on retention in a glass-fiber and -3 -1 Phaeopigment (mg.m ) =fa. ((r. Ra)- R) • s • e . V membrane filter and on the reliability of two storage methods. Arch. Hydrobiol. Beih. Ergebn. = fluorescence reading after acidifi­ Limnol., 14, 46-51. cation 5. Lorenzen, C.F. 1967. Determination of chlorophyll = ration R/R obtained from an extract r a and phaeopigments: spectrophotometric equa­ free from phaeopigment tions. Limnol. Oceanogr. 12: 343-346. fa = f . r .. ( r-1) -l 6. Nusch, E.A. 1980. Comparison of different methods f, R, s, e, V = see above for chlorophyll and phaeopigment determination. Arch. Hydrobiol. Beih. Ergebn. Limnol., 14, 14-36. k) Reproducibility 7. Nusch, E.A. 1984. Ergebnisse eines Ringversuches zur Chlorophyll - a - Bestimmung im Wasser. Every worker should check the reproducibility of the z. Wasser-Ab-Wasser-Forsch., 17, 89-94. method at least once a year by doing 10 replicate 8. Wintermanns, J.F.G.M. & A. de Mots 1965. samples. The value of the coefficient of variation Spectrophotometric characteristics of chloro­ phyll a and b and their phaeophytins in (CV %) should be stated for each pigment. ethanol7 Biochiiiiica et Biophysic a Acta, 109, 448-453. 1) Data reporting

For the data reporting see Sections D.lO and D.ll. 3. Phytoplankton

Any deviation from the procedure recommended here a) Sampling design should be stated on the Plain Language Record when reporting the data. For the purpose of quantitative studies, an integrated sample from the uppermost 10 m shall be obtained by m) References in Chapter D.2. pooling equal volumes of discrete water samples taken with a water sampler from 0-1 * m, 2.5 m, 5 m, 7.5 m and 1. Arvola, L. 1981. Spectrophotometric determination 10 m. If possible, a true integrated sample can be made of chlorophyll a and phaeopigments in ethanol instead using a tube sampler (Willen 1986, Ramberg extraction. Ann.-Bot. Fennici, 18, 221-227. 1976). 2. Jeffrey, S.W. and G.F. Humprey 1975. New spectrophotometric equations for determining chlorophylls a, b, c 1 and c 2 , in higher plants, algae and natural phytoplankton. Biochem. Physiol. Pfl., 167, 191-194.

3. Jespersen, A.-M. & K. Christoffersen 1987. *) Blue-green algae may concentrate close to the Measurements of chlorophyll a from surface. Care should be taken to ensure that these phytoplankton using ethanol as extraction blue-green algae, become represented in the solvent. Arch. Hydrobiol. 109, 445-454. integrated sample. -24- -25-

The integrated sample (independent of the sampling b) Preservation and storage of samples design) should be thoroughly mixed in a large bucket. A first-level subsample of 200 ml is drawn from the Qualitative samples well-mixed sample for quantitative phytoplankton counts. A second-level subsample is drawn for chloro­ Net samples should be preserved immediately with about phyll-~ analysis (see Section D.2). 5 ml cone. formalin per 100 ml. If coccolithophorids are present, 5-10 ml hexamine-neutralized formalin A third-level subsample might be drawn for measurements should be used if the coccoliths need to be preserved. of the irradiance-primary production relation (see Any glass container with a tight screw cap will do, Section D.l.b)). preferably a rather wide-necked one. Plastic containers are not recommended, since some plastics may be Other determinands (e.g. nutrients) might also be permeable to formaldehyde vapour. The samples should be determined from the integrated sample. stored in the dark at room temperature, since formalc;'!ehyde may polymerize to a white precipitate of Additionally, two quantitative samples are recommended: paraformaldehyde at low temperatures. Properly stored one just above and one below the halocline. samples will deteriorate only slowly over a period of several years. Weakly silicified diatom frustules are It is recommended that a net sample be taken from the dissolved first. 0-10 m water column for qualitative study of the microplankton fraction. A plankton net made of gauge Subsamples to be studied alive can be kept fresh for a with 10-20 (-25) 11m meshes, with a mouth diameter of few hours in an open container in a refrigerator. If 15-20 em, a cylindrical part 25-30 em long, and a water samples are to be processed alive according to tapering tail end 15-20 em long will work quite the relevant parts of Annex I of this Chapter, it satisfactorily. should be done as soon as possible. They can be kept unpreserved for up to 24 h in 0.5-1.0 1 polypropylene If net samples are used for biometric measurements, it or polyethylene bottles at in situ temperature if it is should be noted that for species with the smallest below +10° C or 1 month in a deep freeze. Warmer dimension around or below the size of the meshes, cells samples should be refrigerated. towards the lower size range tend to be under­ represented. Therefore, the quant.i tati ve samples have Quantitative samples to be used for biometric measurements of these and smaller species. Samples for quantitative phytoplankton counts should be preserved immediately with 0.5-1,0 ml acid Lugol's solution per 200 ml subsample. If coccolithophorids need to be preserved with the coccoliths intact, a parallel subs ample should be fixed with 0. 5-l. 0 ml alkaline Lugol's solution or 4 ml neutralized formalin. Clear, colourless 2 00 ml glass bottles with tightly

'i >llUlMA ··:.;.;;;;;:s:m,'kw·.m·· ______...,...... ,..,...... , ______,._------

-26- -27-

fitting screw caps should be used for iodine-preserved Analytic grade formalin is to be preferred. Still material. With such bottles it is easy to see when the better is a solution freshly made from paraformal­ iodine becomes depleted and more preservative needs to dehyde. Commercial formalin may contain up to lS % be added. methanol as a stabilizer, and will not give as good fixation. Filter after one week if the solution forms Plastic containers should not be used. Formalin should precipitate or becomes turbid. Add 4 ml per 200 ml not be added to samples preserved with Lugol's solution water sample. Net samples require S-10 ml per 100 ml. unless the storage period is going to exceed one year. c) Qualitative determinations The samples should be counted as soon as possible, preferably within half a year. They should be stored General dark and cool (ca. +4° C). Samples stored for more than one year are frequently of little use. While quantitative investigations have a long standing, and are. performed routinely by most laboratories, the Preservatives greatest weakness has proved to be the identification of the organisms. Acid Lugol's solution (Willen 1962): 1000 ml distilled or deionized water It is becoming more and more evident that the taxo­ 100 g potassium iodide (KI) nomical composition of phytoplankton and the species SO g resublimated iodine (I2 ) succession have considerable value as indicators of the 100 ml glacial acetic acid (cone. CH 3COOH) trophic status of a water mass. Especially the nano­ plankton (2-20 flm) and picoplankton ( < 2 flm) fractions Mix in the order listed. Make sure the previous need much more attention than they have hitherto ingredient has.. dissolved completely before adding the received. next one. Store in a tightly stoppered glass bottle at +4° c. Add O.S-1.0 ml per 200 ml sample. Quantitative phytoplankton counting is a labour­ intensive procedure, and the cost-benefit of the Alkaline Lugol's solution resulting data is questionable. Investing less effort (modified after Utermohl 19S8): in the actual counting process, and more in cheap chlorophyll-a determinations and skill-requiring survey Replace the acetic acid of the acid solution by SO g of the floristic content of samples would apparently sodium acetate (CH COONa). Use a small part of the 3 give more useful results, provided the taxonomic water to dissolve the acetate. Other comments as above. knowledge of the analyst is good enough.

Neutralized formalin (Throndsen 1978): SOO ml cone. formalin (37-40 % aqueous HCHO solution) 500 ml distilled or deionized water 100 g hexamethylenetetramine (CH 2 ) 6N4 (hexamine) ,.,.,,,,..-..., ______------

-28- -29-

Determination Settling procedure

Because of the limitations of the Utermohl technique Before sedimentation the sample should be adapted to and the inverted microscope, accessory techniques are room temperature for about 24 hours to avoid excessive needed for the determination of a number of phyto­ formation of gas bubbles in the sedimentation chambers. plankton species and groups, especially when as Gas bubbles will adversely affect sedimentation, the complete as possible species lists are required. Some distribution of cells in the bottom-plate chamber, and of these non-obligatory methods are reviewed in Annex I microscopy. of this Chapter. Immediately before the sample is poured into the Net samples are recommended to be taken routinely for combined sedimentation chamber, the bottles should be the investigation of sparsely occurring microplankton shaken firmly but gently in irregular jerks to homo­ species with a standard research microscope. The genize the contents. Too violent shaking will produce a advantages include potentially higher resolution, lot of _small bubbles which may be difficult to elimin­ thinner preparations and the possibility to turn the ate. If the sample must be shaken vigorously in order cells around by tapping the cover glass. This is to disperse tenacious clumps, this should not be done especially helpful when examining the plate structure later than one hour before starting sedimentation. of dinoflagellates. The chambers should be placed on a horizontal surface d) Quantitative determinations (phytoplankton counting) and should not be exposed to temperature changes or direct sunlight. Covering the settling chamber(s) with Introduction an overturned plastic box will provide a fairly safe and uniform environment for sedimentation. If moistened The recommendation is based on the counting technique tissue paper is included under the hood, problems with an inverted microscope as described by Utermohl caused by evaporation will be reduced considerably. It (7). In order to elucidate sources of error caused by is essential that the supporting surface is vibration­ subsampling and the settling and counting of free, since vibrations will cause the cells to collect subsamples, the practical application of the method is in ridges. emphasized. To make phytoplankton counting less time­ consuming and the results more comparable, a simplified Settling time is dependent on the height of the chamber procedure along the lines proposed by Willen (11) has and the preservative (e.g. 1, 4). The times given below been adopted. are recommended as minimum. If vibration caused e.g. by traffic is a problem, the minimum time should not be Statistically settling and counting represent two more significantly exceeded. Otherwise it is suggested that subsampling stages (unless the preserved subsample is counting be p'erformed within four days. Sedimented counted in toto, which is unrealistic), which reduce samples not counted within a week should be discarded. the precision of the population estimates (cf. 8, 9). Separated bottom chambers not counted immediately should be kept in an atmosphere saturated with humidity. -30- -31-

Settling time (h) CELL: All non-colonial unicellular species Volume of Height of Lugol's Neutr. Dinobryon Uroglena (disintegrated colonies) chamber ( rnl) chamber (ern) solution formalin Aulacosira 2 1 3 12 Chaetoceros Detonula 10 2 8 24 Leptocylindrus 50 10 24 48 Melosira Skeletonerna (and other chain-forming 100 20 48 48 diatoms) Planktonerna (and other filamentous green algae) One hundred rnl chambers should be used with caution since convection currents are reported to interfere COLONY: Aphanothece Coelosphaeriurn with the settling of plankton in chambers taller than Gornphosphaeria five times their diameter (2, 1). Such chambers can be Microcystis (incl. Aphanocapsa) Gloeotrichia used only when phytoplankton is very sparse, as in late Uroglena (when colonies well preserved) autumn and winter. Halosphaera Sphaerocystis (and similar genera)

If the cells are too strongly stained by iodine for COENOBIUM, with a± fixed number of cells (n): Eudorina (32) comfortable identification, surplus iodine can be Pandorina (16) chemically reduced to iodide by dissolving a small Coelastrurn (8, 16) Crucigenia (4) amount of sodium thiosulfate (Na s • 5 H 0) in the 2 2o3 2 Micractiniurn (4) aliquot to be sedirnented. Pediastrurn (4, 8, 16, 32, etc.) Scenedesrnus (2, 4, 8)

Counting procedure SOME COLONIAL ALGAE are most conveniently counted as groups of four cells, e.g.: Chroococcus In order to save time and to achieve a reasonable Merisrnopedia Crucigeniella accuracy in counting, the sedirnented sample should Dictyosphaeriurn first be examined for general distribution of cells on FILAMENTS to be counted in lengths of 100 ~rn: the chamber bottom, and the abundance and size Achroonerna distribution of the organisms. All species found should Anabaena Anabaenopsis be listed. If the distribution is visually uneven, Aphanizornenon one-sided or in ridges, indicating convection or vi­ Beggiatoa Lyngbya bration, respectively, the settled sample should be Nodularia discarded. If this occurs consistently, measures should Oscillatoria (etc. filamentous blue-green algae) be taken to eliminate the sources of disturbance. The following combinations of objectives and oculars How much of the chamber area should be counted and the are recommended for quantitative microscopy with the magnification to be used is dependent on the size of inverted microscope: the organisms and their abundance, and on the kind of counting units used. The following counting units are recommended: -32- -33-

Microplankton> 20 [lm Nanoplankton 2- 20 [lm Count 95 % C.L. ( %) Picoplankton <2 [lm 4 100 Objective lOx or 16x or 25x 40x or 63x 5 89 7 76 Oculars 12.5-16x 10-12.5x 10-12.5x 12.5-16x 10-12.5x 10 63 15 52 20 45 Small microplankton species can preferably be counted 25 40 40 32 together with the nanoplankton when they occur in 50 28 abundance, or they can be counted using an objective 75 23 100 20 with intermediate magnification, 20-25 x. A grid of 5x5 200 14 squares in one of the oculars is very helpful when 400 10 500 8.9 counting dense fields of small cells. 700 7.6 1000 6.3 2000 4.5 Counting should normally be restricted to the 10-12 5000 2.8 most abundant species. When the diversity of a sample 10000 2.0 is high, however, up to 15 species may have to be It should be recognized that these are not maximum counted, and during bloom conditions probably less than errors. The statistics assume perfectly random distri­ ten will be enough. According to Willen (11) 90 per bution of cells on the bottom of the sedimentation cent or more of the total biomass was made up by six chamber, a condition which is probably never realized. species in nutrient-rich, and by 7-8 species in The several subsampling steps involved also tend to nutrient-poor, lake waters. increase the variance (cf. 9, 10).

At least 50 counting units of each taxon should be With species for which the counting unit is smaller counted, and the total count should exceed 500. Fewer than the individual, e.g. some colonial forms, chain­ counted units will progressively decrease the precision forming diatoms, and filamentous species with average of the count and increase the statistical error of the filament length in excess of 100 [Am, the distribution population estimate. The approximate 95 % confidence of the counting units will be aggregated even in limits of a selected number of counted units are given perfectly sedimented samples. The variance will be below. They have been calculated according to the higher, and the precision accordingly lower. If it is formula: necessary to keep the error within the same limits as for "randomly" distributed units, the number of counted 95 % C.L. = n units should be increased in the ratio average size of individual/size of counting unit.

where n is the number of units counted. (Actually the At low magnification, a sufficient number of crossed error is not symmetrical, but increasingly asymmetrical transects should be counted to reach the required with lower counts. Thus, for four units counted the number of counting units. At high magnification, fields theoretical limits are -73 to +156 %.) -34- -35- of view evenly spaced along crossed transects should be protozooplankton are a separate group and must not be used. At intermediate magnifications either method can mixed with the phytoplankton. Thus, they must not be be employed, depending on the density of the units to included in abundance or biomass values of phyto­ be counted. Considering the many other sources of error plankton. involved in the method, it is unnecessary to spend time on randomization of the fields to be counted. Only Cleaning of the sedimentation chambers exceptionally will it be necessary to count the whole chamber bottom for very large cells, such as Ceratium After use no part of the combined sedimentation chamber and Coscinodiscus. should be allowed to dry out before it is carefully cleaned. Dried phytoplankton or formalin preservative The number of counting units per dm 3 sea water is may be quite difficult to remove. The separate parts calculated by multiplying the number of units counted are first rinsed under running tapwater, then soaked with the coefficient C, which is obtained from the for a few minutes in luke-warm water with some non­ following formulas: abrasive detergent added, then cleaned with a soft brush or soft tissue paper, and rinsed with tapwater. 1000 or A . 1000 Finally they are given a rinse with deionized or dis­ c = A c = a • V tilled water, and are put away to dry. Special care N al . v 2 should be taken not to scratch either end of the top where cylinder and the entire upper surface of the bottom A = cross-section area of the top cylinder of the plate. combined sedimentation chamber; the usual inner diameter is 25.0 mm, giving A = e) References in Chapter D.3. 491 000 000 ~m' (the inner diameter of the 1. Hasle, G.R. 1978. The inverted-microscope method. bottom-plate being irrelevant) In: Sournia, A. (ed.): Phytoplankton manual. N = number of counted fields or transects UNESCO Monogr. Oceanogr. Method. 6: 88-96. Paris. al = area of single field or transect = total counted area 2. Nauwerck, A. 1963. Die Beziehungen zwischen a2 zooplankton und Phytoplankton im See Erken. 3 v = volume (cm ) of sedimented aliquot Symb. Bot. Ups. 17(5): 1-163.

3. Ramberg, L. 1976. Relations between phytoplankton Reliable quantitative counting of the picoplankton and environment in two Swedish forest lakes. fraction requires fluorescence microscopy (cf. Annex I Scripta Limnologica Upsaliensis 426, Limno­ logical Inst., Univ. Uppsala. of this Chapter). 4. Rott, E. 1981. Some results from phytoplankton counting intercalibrations. Schweiz. S. When counting phytoplankton in a sedimentation chamber, Hydr~l. 43: 34-62. it is suitable to count protozooplankton (e.g. ciliates 5. Throndsen, J. 1978. Preservation and storage. In: and colourless flagellates). This recommendation is Sournia, A, ( ed.) : Phytoplankton manual. also valid for these forms (cf. Annex I of this UNESCO Monogr. Oceanogr. Method. 6: 69-74. Paris. Chapter). However, it must be stressed that the -36- -37-

6. Utermohl, H. 1931. Neue Wege in der quantitativen Butcher, R.W., 1959-1967. An introductory account Erfassung des Planktons. Verh. int. Verein. of smaller algae of British coastal waters. Limnol. 5: 567-596. Part I. Introduction and Chlorophyceae. Part IV. Cryptophyceae. Part VIII. Euglenophyceae. 7. Utermohl, H. 1958. zur Vervollkommnung der Fish. Invest. Ser. 4. London. quantitativen Phytoplankton-Methodik. Mitt int. verein. theor. angew. Limnol. 9: l-38. Cleve-Euler, A., 1951-1955. Die Diatomeen von Schweden und Finnland. 1-5. Kungliga svenska 8. Venrick, E.L. 1972. The statistics of subsampling. vetensk.-akad. handl. Ser. 4. Stockholm. Limnol. Oceanogr. 16: 811-818. Cupp, E.E., 1943. Marine plankton diatoms of the 9. Venrick, E.L. 1978a. The implications of west coast of North America. Bull. Scripps subsampling. In: Sournia, A. (ed.): Phyto­ Inst. Oceanogr. Univ. Calif. La Jolla. 5(1). plankton manual. UNESCO Monogr. Oceanogr. London. Method. 6: 75-87. Dodge, J.D. 1982. Marine dinoflagellates of the 10. Venrick E.L. 1978b. How many cells to count? In: British Isles. Her Majesty's Stationery Sournia, A. (ed.): Phytoplankton manual. Office, London. UNESCO Monogr. Oceanogr. Method. 6: 167-180. - 1985. Atlas of Dinoflagellates. Farrand Press, London. 11. Willen, E. 1976. A simplified method of phyto­ plankton counting. Br. phycol. J. 11: 265-278. Drebes, G., 1974. Marines Phytoplankton Eine Auswahl der Helgolander Planktonalgen 12. Willen, E. 1985. Vaxtplankton Rakning och (Diatomeen, Peridineen). 1. Aufl. Stuttgart. volymbestamning. In: Recipient kontroll vatten Metodunderlag. Naturvardsverket Ettl, H. 1978. Xanthophyceae. 1. Teil. Sliss­ Rapport 307 5, National Environmental wasserflora von Mitteleuropa 3: 1-530. Protection Board, Solna, Sweden, 18 pp. - 198 3. Chlorophyta I. Phytomonadina. Sliss­ wasserflora von Mitteleuropa 9: 1-807. 13. Willen, T. 1962. Studies on the phytoplankton of some lakes connected with or recently isolated Fott, B., 1959 and 1971. Algenkunde. Jena. from the Baltic. Oikos. 13: 169-199. - 1972. Chlorophyceae (Grlinalgen). Ordnung: Tetrasporales. Die Binnengewasser 16, Das Phytoplankton des Slisswassers 6: 1-116, pl. 1-47. f) Identification literature Geitler, L., 1925. Cyanophyceae. Die Slisswasser­ flora Mitteleuropas. 12. Leipzig. The following list is not complete. Articles concerning - 1932. Cyanophyceae. Rabenhorst Krypto- single or only a few species have not been included. gamenflora von Deutschland, Osterreich und der Schweiz. 14. Leipzig.

Phytoplankton identification literature Gran, H.H., 1905. Nordisches Plankton. Botanischer Teil. 19. Kiel. Balech, E., 1976. Some Norwegian Dinophysis species (Dinoflagellata). Sarsia, 61. Hendey, N.I., 1964. An introductory account of the smaller algae of British coastal waters. 5. Bourrelly, P., 1966. Les algues d'eau doL 1' . Bacillariophyceae. Fish. Invest. Ser. 4. Les algues vertes. Paris. London. - 1968. Les algues d' eau douce. 2. Les jaunes et brunes. Paris. Huber- pestalozzi, G., 1938-1961. Das phyto­ - 1970. Les algues d' eau douce. 3. Les algues plankton des Slisswassers. Die Binnengewasser. bleues et rouges. Paris. 16: 1. Blaualgen, Bakterien, Pilze. 2. Chryso­ phyceen, fqrblose Flagellaten, Heterokonten, Diatomeen. 3. Cryptophyceen, Chloromonadinen, Peridineen. 4. Euglenophyceen. 5. Chloro­ phyceae - Volvocales. Stuttgart. -38- -39-

Hustedt, F., 1930. Bacillariophyta (Diatomeae). Smith, G.M., 1920-1924. Phytoplankton of the Die Slisswasserflora Mitterleuropas. 10. Jena. inland lakes of Wisconsin. 1-2. Wisconsin - 1930-1966. Die Kiselalgen. Rabenhorst Krypto­ geological and natural history survey gamenflora von Deutschland, Osterreich und der bulletin. 57. Madison. Schweiz. 7.1-.3. Leipzig. Sournia, A. 1978. A guide to the literature for Jlrnefelt, H., Naulapll, A., Tikkanen, T., 1963. identification. In: Sournia, A. (ed.): Plankton-opas. Helsinki. Phytoplankton manual. UNESCO Monogr. Oceanogr. Methodol. 6: 154-159. Komarek, J. & Fott, B. 1983. Chlorophyceae (Grlinalgen). Ordnung: Chlorococcales. Die Starmach, K., 1966. Cyanophyta, Glausophyta. Flora Binnengewlsser 16, Das Phytoplankton des slodkowodna polski. 2. Warszawa. Slisswassers 7(1): 1-1044. - 1968. Chrysophyta 1. Chrysophyceae. Flora slodkowodna polski. 5. Warszawa. Komark, J. & Ettl, H., 1958. Algologische Studien. - 1968. Chrysophyta 3. Xantophyceae. Flora Prag. slodkowodna polski. 7. Warszawa. - 1972. Chlorophyta 3. Ulotrichales, Ulvales, Lebour, M., 1925. The dinoflagellates of northern Prasiolales, Sphaeropleales, Cladophorales, seas. Plymouth. Chaetophorales, Trentepholiales, Siphonales, Dichotomisiphonales. Flora slodkowodna polski. Pankow, H., 1976. Algenflora der Ostsee II. 10. Warszawa. Plankton. Jena. - 1974. Pyrrophyta. Cryptophyceae, Dinophyceae, Raphidophyceae. Flora slodkowodna polski. 4. Warszawa. Peters, N., 1930. Peridinea. In: Grimpe & Wagler (Ed.), Tierwelt der Nord- und Ostsee. II. - 1985. Chrysophyceae und Haptophyceae. Sliss­ wasserflora von Mitteleuropa 1: 1-515. Reinhardt, P., 1972. Coccolithen. Kalkiges Plankton seit Jahrmillionen. Wittenberg Tikkanen, T. 1986. Kasviplanktonopas. 278 pp. Lutherstadt. Helsinki. Schiller, J., 1933-1937. Dinoflagellatae. Rabenhorst Kryptogamenflora von Deutschland, Osterreich und der Schweiz. 10:3. Leipzig. Phytoplankton checklists Simonsen, R. 1987. Atlas and Catalogue of the Diatom types of Friedrich Hustedt. In the BMP the Preliminary Check-list of the Phyto­ Volume 1. Catalogue. Volume 2. Atlas, Plates 1-395, op.2-106 plankton of the Baltic Sea (Section D.4.) shall be Volume 3. Atlas, Plates 396-772, op.l07-175 used. Additional information can be found in the Skuja, H., 1934. Beitrag zur Algenflora Lettlands. following check-lists: I. Acta Horti. Bot. Univ. Latviensis. 7:1932. Riga. Christensen, T. & Thomsen, H .A., 1974. 1948. Taxonomie des Phytoplanktons einiger Algefortegnelse. Kobenhavn. Seen in Uppland, Schweden. Symbolae botanicae upsaliensis. 9:3. Uppsala. Christensen, T., Koch, c. & Thomsen, H.A., 1985. - 1956. Taxonomische und biologische Studien Distribution of algae in Danish salt and tiber das Phytoplankton schwedischer Binnen­ brackish waters. 64 pp. Copenhagen. gewlsser. Nova acta regiae societatis scientiarum upsaliensis. Ser. 4. 16:3. Drebes, G. & Elbrlchter, M., 1976. ~Checklist of Uppsala. Planktonic Diatoms and Dinoflagellates from - 1964. Grundzlige der Algenflora und Alpenflora Helgoland and List (Sylt), German Bight. der Fjeldgegenden urn Abisko in Schwedisch­ Botanica Marina. 29. Lappland. Nova acta societatis scientiarum upsaliensis. Ser. 4. 18:3. Uppsala. -40- -41-

Edler, L., HiHlfors, G. and Niemi, A., 1984. A g) Biomass transformations preliminary check-list of the phytoplankton of the Baltic Sea. Acta Bot. Fennica 128: 1-26. Introduction Heimdal, B.R., Hasle, G.R. and Throndsen, J., 1973. An annotated checklist of plankton algae from the Oslofjord, Norway (1951-1972). Norw. Depending on the purpose of the investigation, J. Bot. 2 0. phytoplankton biomass can for example be expressed as Hendey, M.I., 1974. A Revised Checklist of British cell volume (weight), plasma volume or carbon. The Marine Diatoms. J. Mar. Biol. Ass. U.K. 54. transformations to cell volume rely on measurements of HiHlfors, G., 1979. A preliminary check-list of the size of the species, and a large number of shapes the phytoplankton of the northern Baltic Sea. have to be used for the different organisms. The National Board of Waters, Helsinki. transformation of cell volume to plasma volume includes Parke, M. & Dixon, P.S., 1976. Checklist of an estimate of the vacuole volume, and the calculation British Marine Algae - Third Revision. J. Mar. Biol. Ass. U.K. 56. of cell carbon is in turn based on the plasma volume. Zetterberg, G. 1986. Phytoplankton Code List P4, The focrmulas recommended below are a step towards a Version 86165-GUZ. Code Centre, Swedish Museum more uniform treatment of counted organisms, in order of Natural History. to get comparable results from different parts of the Protozooplankton identification literature Baltic.

Bock, K.J., 1967. Protozoa. Fich. Ident. Plancton. Cons. int. Explor. Mer, 110, 4 pp. Stereometric shapes and formulas for common phyto­ plankton and protozooplankton in the Baltic Brandt, K. & Apstein, C., 1964. Nordisches Plankton. Zoologischer Teil. 7. Protozoa. Kiel. Volumes of species with a small size variation can be Corliss, J.O. 1979. The ciliated protozoa. calculated as annual median values. For species present Characterization, Classification and Guide to only seasonally, median values from several years can the Literature. 2. ed. Pergamon Press. Oxford. be used. Species with a large size variation are Grimpe, G. 1940. Die Tierwelt der Nord- und suitably divided into size groups during counting (e.g. Ostsee. Akademische Verlagsgesellschaft, Leipzig. diatoms and monads).

Kahl, A., 1935. Protozoa. I. Ciliata. In: Die Tierwelt Deutschlands. begr. von F. Dahl. Methods for calculating volumes of different plankton Jena. species are given in Table D.2. Kofoid, A. & Campbell, T., 1929. Conspectus of the Tintinnoinea. Univ. of Calif. Publ. in For comparison of the volume of different species in Zoology. 34. - 1939. The Tintinnoinea. Bull. of the Mus. of some parts of the Baltic Sea reference is made to ( 1, Comp. Zoo!. at Harvard Coll. 84. 2, 3, 4, and 12). Schwarz, s. 1964. Die Tintinnoidea. Hydrobiologia 23.

4 483184A -42- -43-

TABLE 0.2. Stereometrical formulas for common phytoplankton taxa in the Baltic

CONE

STEREOMETRIC FORMULAS USED

CYLINDER

() () TRUNCATED CONE

2 2 SPHERE 2 2 l•h•(d +d d +d ) l•h• (R +Rr+r ) l 1 2 2 3 1 2 n • d' ~ -6- ~ TRAPEZOID ROTATIONAL ELLIPSOID WITH ELLIPSE-SHAPED CROSS-SECTION d b

I h•d•(a+b) h 2 - ID•

ROTATIONAL ELLIPSOID WITH CIRCULAR CROSS-SECTION SPECIES

x obligate heterotrophic xx facultative heterotrophic 9l 1 autotrophic/heterotrophic unknown NOSTOCOPHYCEAE (Cyanophyceae)

ELLIPSOID x Achroonema! cylinder Anabaena: cylinder Aphanizomenon: cylinder 0. Chroococcus: cells as spheres, dividing cells as spheres/2 d Gomphosphaeria: outer sphere - inner sphere

PARALLELEPIPED

rt-----nI b -44- -45-

[yY!.qbya. lir.:neti,:-a: cylinder :'·fe"t'1;smopedi:::.: cells as spheres, dividing cells as spheres/2 ? CochZ.odinium: rotat\·onal ellipsoid :'1iarocystis: cells as spheres (cf. Reynolds & Jaworski, 1978) Dinophysis acuminata: two truncated cones - 50% :Vodu.laria: cylinder norvegioa: " Jsc·iZlatoria: cyLinder

P~onmidium: cylinder

Pseudoanabaen~: cylinder

CRYPTOPHYCEAE x Chilomonas: cone+ sphere/2, rotational ellipsoid (ellipse-shaped x Dinophysis rotundata: cylinder cross-section) Chroomonas: cone + sphere/2, rotational ellipsoid (ellipse-shaped cross-section) Cryptomonas: rotational ellipsoid (ellipse-shaped cross-section)

Rhodomonas: cone + sphere/2

DINOPHYCEAE Dinophysis acuta: ellipsoid . . d 2 XX Amphidinium: rotational elltpsot · j (ellipse-shaped cross-section)

Cerotiwn fu'!'ca: cone + parallelepiped + trapezoid + cylinder lineatum: -c::::::D ';------ro ' X Ebria: sphere/2 - 307.. If the skeleton is missing calculate the <:D "t:!===:::J volume as a sphere XX Glenodinium: sphere - 10 to 407. Gonyaulax caterr.ata: sphere/2

Ce~atiwn fusus: cylinder + truncated cone Gonyau~ diacantha: trapezoid + parallelepiped/2 t:riacantha:

Ceratium longipes: macrocer-=>s: cone + cylinder + sphere/2 -46- -47-

CHRYSOPHYCEAE Gor:ya:.-~ lax grindleyi: polyedra: sphere x Bicocoeca: tc.:narens-i.s : Dinobryon: rotational ellipsoid or sphere !Jt:;niodoma: sphere Kephyrion: xx Gymnod.;ni:,vn spp. : Dictyocha: cylinder 2 cones or 2 sp~eres/2 - 10 to 40% Cy't'odiniwn spp.: Gymnodinium simplex: 2 spheres/2 Gymnodinium splendens: (sphere/2 + cone)/2 .::eteroocapsa triquetra: 2 cones - 207. h Min:...scula bipes: cone - 507. x Noctiluca mill-aris: sphere xx Oxytoxum: 2 cones Polykrikos: cylinder Prorocentrum Oalticum: sphere- 10%, measurement should be made MaZZ.cmona~: cone + sphere/2 at maximum diameter Symqc: cone + sphere/2 Prorocentrum micans: parallelepiped+ parallelepiped/2 Vrog Z.tJI".a: FTotoperidinium: General formulas for volume calculations: sphere or rotational ellipsoid (ellipse-shaped UrcgtlJnopsis: cone + sphere/2, 2 cones, sphere, 2 cones + cross-section) cone. Generally the volumes have to be reduced by 10 to 30% because of the concavity of the ventral side. BACILLARIOPHYCEAE x P. :.:reve: Aohnanthes: parallelepiped X brevipes: ActinoayoZ.us: cylinder cone + sphere/2 X co:'l.icoides: Amphipro~: ellipsoid X steinii: Amphora: ellipsoid X cl~~dicans: (cone + 2 cones) - 30% Aster·ione lZa: parallelepiped

X conicum: (cone + 2 cones) - 257. Attheya: ellipsoid

X depPesswn: (cone + 2 cones) - 307. Baciltaria: parallelepiped

X di~ergens: (cone+ 2 cones) - 307. Biddulphia: .ellipsoid

X J'Nlnii: (cone + sphere/2) - 20% Cerataulina: cylinder

X ?allidum: (cone + sphere/2) - 30% Chaetooeros: ellipsoid

X pellucidum: (cone+ sphere/2) - 10% Cocooneis: ellipsoid Scrippsiella faeroense: sphere Cosoinodisous: cylinder + two caps

? ~arnouia: rotational ellipsoid Cymbe!la: ellipsoid DaotyZ.iosolen: cylinder Detor.uZa,: cylinder ~iatoma: parallelepiped DipZoneis: ellipsoid Ditylwn: cylinder ·, "'""'~-,,------~------·

-48- -49-

Spithemia: ellipsoid

£~notia: ellipsoid PRAS!NOPHYC£AE ?~gila~;a: parallelepiped Gomphonema: ellipsoid Pyra:nimonas: trapezoid: Gl'amr.la.taphom: parallelepiped

Gui~nrdia: cylinder CHLOROPHYCEAE Gyrosigma: ellipsoid Ankistrodesm~s: cone+ cone Hantzschia: cylinder Carteria: sphere or rotational ellipsoid Lauderia: cylinder Chlamydomonas: sphere or rotational ellipsoid Leptocy[indrus: cylinder Chlorel~: sphere Licmophora: parallelepiped/2 Chlorogonium: cone + cone .''4elosira: cylinder Dich~yospha~rium: cells as spheres Navicula: parallelepiped, parallelepiped/2 Golenkinia: sphere Nitzschia: parallelepiped, parallelepiped/2 Kirchneriel~: cone+ cone or rotational ellipsoid

Pinnularia: parallelepiped, parallelepiped/2 Monor~phidium: cone + cone or rotational ellipsoid

Porosira: cylinder Oocystis: rotational ~llipsoid Rhabdonema: parallelepiped Pediastrum: cylinder with height as the shortest side of one cell Rhizosolenia: cylinder in the middle of the coenobium Rhoicosphenia: parallelepiped/2 Rhopalodia: ellipsoid Skeletonema: cylinder Stephnnodiscus: cylinder Surirella: ellipsoid Synedra: parallelepiped, parallelepiped/2

TaOellaria: parallelepiped h T.iulassionema: parallelepiped

Thalassio3i~a: cylinder P~nctonema: cylinder

XANTHOPHYCEAE Scenedesmus: cells as rotational ellipsoid or cone+ cone

Centritractus: rotational ellipsoid XX Monads size groups 2 - .(3 )lm 3 -<5 )lffi Gloeochloris: cells as spheres or rotational ellipsoids 5 -<7 )lm sphere or rotational ellipsoid Ophiocytium: cylinder 7 -< 10 )lm 10 -< 15 )lffi EUGLENOPHYCEAE 15 -< 20 )lm, etc.

Colacium: rotational ellipsoid P!COPLANKTON < 2 )lm E:uqZena: CHOANOFLAGELL!DA Eu trep tia : cylinder + cone, cone or rotational ellipsoid Et< treptie Lla: Acal':thoeaa: Trachelomonas: sphere or rotational ellipsoid Aoanthoecopsis: CalZiacantha: rotational ellipsoids or spheres Crinolina:

Desmarel~: -50- -51-

Plasma volume calculations

All phytoplankton, except diatoms.

Dia.phcx~oeca: In this recommendation it is assumed that plasma volume Dipiotheca: (PV) equals cell volume (CV). See, however, Sicko-Goad Mor;osiga: Parvic:o't'bicula: et al. ( 5) . Pleu.rasiga: rotational ellipsoids or spheres Salpingoeca: PV = CV Sa"i'oeca: Stephanoeca: Sa vi Zlea: unit:

CILIATES WITH ENDOSYMB!OTIC ALGAE Diatoms 5 Mesodiniwn < 30 um: 2 · sphere 1 8 > 30 um: sphere Different methods for calculation of the plasma volume 1 have been suggested by (6 and 7). The method CILIATES recommended here is a modification of that given by . l 4 Cl ate: sphere • 3, cone or ellipsoid Strathmann to include pennate diatoms. Plasma volume Didynium: truncated cone Lohmanniella: sphere (PV) equals cell volume (CV) minus vacuole volume (VV).

Str>ombidium conicwn: cone Strombidium lagenula: cone The vacuole volume is calculated by subtracting plasma Strombidium stylifer: cone thickness (c) from the measurements of length, breadth Tiarina: cylinder + cone etc. (see Figure D.l. below). If a measurement extends Tontonia gracillima: sphere Tontonia appendicu l::;_rij'orrnes: 2 truncated cones from cell wall to cell wall, 2 c should be subtracted

Tintinnids from the measured value. When using a measurement such

Coxliaila helix: cylinder + cone as radius that is not bounded by the cell wall and thus Favella denticulata: cylinder +cone contains only one layer of plasma, only c should be Helicostomella subuZata: sphere- 307. subtracted. Values of c have been given by Lohmann (8) Stenosomella nucula: sphere- 30% and Smayda (6) and are in the range of 1 ~m. Stenosomella ventricosa: cone Tintinnopsis beroidea: cone Tintinnopsis tubulosa: cone D•e to the high content of organic matter in the

Volume calculations for J'intinnids may be difficul-: due :;.::' :::re shens of ..~ole, only 90 % of the vacuole volume should be the _organisms. The measurements rrrv.st be do;:.e when ~he :JiZ~:ate iz :::·!1r:.side subtracted from the cell volume to give the plasma the shell. In preserved sa:nples, ho1Jever, the ci~i.:~.te ~s<.1an;,. stays inside volume. the shell. In that case a certai.>; aJ110unt .'n'..tst be su.O~r:zcted frcm the sJ:elZ volume. This amount nrust be j'c!md from meas~{remenr::s of the she~l and ~he ciliate. It is sufficient to measure this in one sample. PV = CV - (0.9 VV)

unit: ~m 3 ~--~:::wp~·'''·'~.------·--"-

-52- -53-

FIGURE D.l. Girdle view of a centric diatom with For comparison of the carbon content of different diameter d, height h and plasma species in some parts of the Baltic Sea, see Smetacek thickness c. (2) and Edler (4).

h) Presentation of results and data reporting

PV • CV - (0.9 • VV) 3 For the data reporting see Sections D.lO and D.ll. unit: /

0 ~ • d' • h Cell Volu~e: -- -- 4 i) References V•cuole Volume: t•{d·2c)'•(h·2c) I h -- 4 1. Melvasalo, T., Viljamaa, H. and Huttunen, M.,

!l~sm• ~olume: CV ~ (0.9 · VV} 1973. Plankton methods in the Water Conser­ I vation Laboratory in 1966-1972. Reports of the ~- ..... f c - - - Water Conservation Laboratory Helsinki. 5. c c 2. Smetacek, V., 1975. Die Sukzession des Phyto­ d plankton in der westlichen Kieler Bucht. Diss. Kiel.

3. McKellar, H. & Hobro, R., 1976. Phytoplankton - Zooplankton Relationships in 100-Liter Plastic Bags. Asko Contr. No. 13.

Carbon content calculation 4. Edler, L. 1977. Phytoplankton and primary production in the Sound. Diss. Goteborg.

Based on the above biomass expressions, it is possible 5. Sicko-Goad, L., Stoemer, E.F. and Ladewski, B.G., to calculate the carbon content of phytoplankton. 1977. A morphometric method for correcting phytoplankton cell volume estimates. Several equations based on both cell volume and plasma Protoplasma 93. volume have been proposed by e.g. Strickland (9), 6. Smayda, T.J., 1965. A quantitative analysis of the Mullin et al. (10), Strathmann (7), Eppley et al. (11). phytoplankton of the 1~lf of Panama II. On the relationship between C assimilation and the diatom standing crop. Inter-Amer. Trop. Tuna. Comm. 9:7. It is recommended that the plasma volume be multiplied 7. Strathmann, R.R., 1967. Estimating the organic by a factor of 0.13 for armoured dinoflagellates (2) carbon content of phytoplankton from cell and 0.11 for all other phytoplankton (7) and ciliates volume or plasma volume. Limnol. Oceanogr. 12.

(2). 0.13 has been chosen because of the higher carbon 8. Lohmann, H., 1908. Untersuchungen zur Feststellung content of the cell walls of armoured dinoflagel_ s des vollstandigen Gehaltes des Meeres an Plankton. Wiss. Abt. Kiel NF. 10 •.

armoured dinoflagellates: C = PV • 0.13 9. Strickland, J.D.H., 1960. Measuring the production of marine phytoplankton. Fish. Res. Bd. all other phytoplankton ~anada, Bull. No. 122. and ciliates: C = PV . 0.11 units: c (carbon) - picogram PV (plasma volume) - ~m' -55- -54-

ANNEX I 10. Mullin, M.M., Sloan, P.R. and Eppley, R.W., 1966. Relationship between Carbon Content, Cell Volume and Area in Phytoplankton. Limnol. Annex I to Section D.3. Phytoplankton Oceanogr. 11.

11. Eppley, R.W., Reid, F.M.H. and Strickland, J.D:H., Notes on accessory methods for species determination, 1970. Estimates of phytoplankton crop s~ze, growth rate and primary production. Bull. and on the determination of protozooplankton vs. non­ Scripps Inst. Oceanogr. 17. photosynthetic phytoplankton. 12. Kononen, K., Forsskahl, M., Huttunen, M., Sandell, M. and Viljamaa, H., 1983: Practical problems Fluorescence microscopy is needed in order to obtain encountered in phytoplankton cell volume calculations using the BMB recommendation in reliable counts of unicellular autotrophic picoplankton the Gulf of Finland. Limnologia 15/2. species. Experienced workers may reach order-of­ magnitude accuracy with relatively large cells such as Micromonas pusilla and Nannochloropsis sp. by the Utermohl technique, but if mineral particles are abundant, the results may be questionable. Coccoid

blue-green algal cells about 1 ~m in diameter or smaller can be distinguished from true bacteria, and thus quantified, only by fluorescence microscopy. Methodological details are given by Hoppe et al. (3), Vargo (8) and Haas (2).

Centrifugation of living water samples is recommended for the study of delicate nanoplankton flagellates whose morphology usually deteriorates upon fixation to the extent that they become virtually unrecognizable (e.g. small naked dinoflagellates, many chrysophytes and prymnesiophytes, some prasinophytes, and most zooflagellates). Besides, several genera or even some species are well characterized by their mode of swimming (e.g. Pseudopedinella, Prymnesium' Chryso­ ehromu lin a' Pavlova, Mieromonas pusi lla , Nephroselmis·) •

A simple bench centrifuge with a swing-out head taking 4-6 25 ml pointed centrifuge tubes is adequate. At 1500-2000 rpm ·a centrifugation time of 10 min is sufficient. ------,------

-56- -57-

To optimize the viability of the cells under the Diatom preparations are necessary for the determination microscope, the glassware should be as clean as of most nanoplanktonic disc-shaped centric diatoms possible (cf. 9). (e.g. CyeLoteLLa, Stephanodiseus and ThaLassiosira) even at the generic level. The majority of littoral Dry preparations observed with a good oil immersion pennate diatoms also require diatom preparations for phase-contrast objective make possible the determination at the species level; for some navi­ determination of a limited number of species in the culoids, even at the generic level. genera Paraphysomonas and Chrysoehromu Lina on the basis of their scale morphology, some coccolithophorids, and A rapid and satisfactory method of making diatom most choanoflagellates. preparations is as follows: Put less than 0. 5 ml of concentrated sample into a test tube. Add ca. 1 ml Suitably concentrated material obtained by centri­ cone. HN0 3 . Hold the test tube with a clothes-peg or fugation as described above is pipetted onto cleaned similar, and heat on a small flame inclining the test cover glasses, fixed by exposure to the vapour of a few tube 45-60" from the vertical and continuously shaking of 1-2 % Oso solution inside a Petri dish small drops 4 with caution in order to avoid overheating and for 30-45 s (under a fume hood) , and allowed to dry. spitting. Keep the opening of the test tube away from The salt is washed away by immersing the cover glasses yourself in a safe direction. Boil for 1-5 minutes into distilled or deionized water for 10 min, after until the solution is clear and white vapours start to which they are dried again. Finally the cover glasses form. If the material is not completely oxidized after are turned specimen side downwards, and are glued to 5 min., add a few drops of cone. H so (beware of 2 4 glass slides. overheating) and repeat.

Electron microscopy is required for a positive The oxidation with acids should be performed under a determination to species level of most scale-covered fume hood. If a fume hood is not available, the acids organisms. These include most prymnesiophytes and can be replaced by enough K s o to make a nearly 2 2 8 prasinophytes, some chrysophytes (Mallomonadaceae and saturated solution at +100" c. The sample volume can be Paraphysomonadaceae), and certain protozoan groups. increased to ca. 2 ml. Heat for 1-2 h in a boiling water bath occasionally replacing the evaporated water. The method of making direct preparations onto specimen grids is very similar to that described for dry Cool and dilute the contents of the test tube by adding preparations. A detailed account of the technique has at least 10 ml water and transfer to a centrifuge tube. been given by Moestrup & Thomsen (5). Species with thin Spin the material down at 1500-2000 rpm for 20 min. organic scales require shadowing for TEM. Most Remove the supernatant preferably using a Pasteur silicified scales can also be determined without pipette attached to a water suction pump. Resuspend the shadowing. Strongly calcified coccolithophorids usually sediment with a jet of water from a wash bottle, and require SEM, as do a number of dinoflagellates with spin it down again for 10 min. Repeat the rinsing pro­ thin thecal plates. cedure at least five ~imes. Use distilled or deionized water in no less than the two last rinses. Dilute with

5 483184A -58- -59- water to a suitable (by experience) turbidity and erally considered by botanists as algal classes within transfer a few drops to a clean cover glass. Evaporate a number of divisions (e.g. 1) are treated by the water, preferably slowly under cover to achieve an zoologists as orders of the class Phytomastigophorea even distribution. Add a sui table amount of embedding (Phylum Sarcomastigophora, Subphylum Mastigophora 1 4) . medium with a high refraction index (e.g. While groups like the choanoflagellates and the Clophenharpiks, Hyrax, Styrax) onto a clean slide, bodonoid flagellates, which until recently were claimed place the cover glass on the top (or pick it up with by phycologists, now seem to be generally accepted to the embedding medium on the slide), and heat gently on belong in the animal kingdom, some groups are still a hot plate to expel air bubbles and evaporate the disputed or uncertain, or lack a satisfactory system­ solvent. atic superstructure, e.g. the bicosoecoids and many colourless members of the Proto- and Polyblepharidinae It should be recognized, however, that some of the and Polymastiginae ( 6, 7), etc. On the other hand, smallest centric species, and small species of the connections have been found in the Pedinellales ( 9) genera Fragi Laria, Achnanthes. and NavicuLa, at least, between a chrysophyceaen lineage and members of the and some Nitzschia species, still require electron protozoan order Actinophryida (Phylum Sarco­ microscopy (TEM or SEM) for a safe determination. mastigophora, Class Heliozoea). There are also a few true animals which function as plants because they have Protozoa and non-photosynthetic phytoplankton intracellular photosynthetic symbionts (e.g. Mesodinium). When working with phytoplankton, it is convenient to determine the protozooplankton as well. The distinction Even with the procaryotes, a uniform conception is between algal and protozoan higher systematic units is lacking regarding the limits of the systematic entities vague to the extent that some researchers tend to and their names. The blue-green algae seem to be a classify them together as protists. Most genuine algal rather homogeneous group, regardless of whether they classes contain species or even whole genera which are are referred to as Cyanophyceae, Nostocophyceae or non-photosynthetic, and which thus functionally should Cyanobacteria. Problems of delimitation arise when be regarded as zooplankton, for example, Cryptophyceae: morphologically similar non-photosynthetic types are CryptauLax, Cya t homonas, KatabLepharis, Leucocryptos1 included (e.g. Beggiatoa, Achroonema), or when the p .p., Dinophyceae: Amphidinium p •P • I Gymnodinium organisms are so small, i.e. usually less than 1-1.5 ~m Gyrodinium p.p., Oxyrrhis, NoctiLuca' ObLea' wide, that special methods (e.g. fluorescence Protoperidinium, Ebria 1 Prymnesiophyceae: Ba Laniger 1 microscopy) are required to determine whether the cells Chrysophyceae: Paraphysomonas1 Euglenophyceae: Astasia.' contain chlorophyll or not. This concerns for instance Peranematales1 Chlorophyceae: PoLytoma. narrow OsciLLatoria spp. vs. Achroonema spp., Merismopedia wa,rmingiana vs. Lampropedia hya tina , and A major obstacle to a uniform treatment is the distinc­ very small photosynthetic coccoid cells vs. true tion between the Botanical and Zoological Nomenclatural bacteria. Codes and the incompatibility of the systematic super­ structures. A number of different phyletic groups gen- -60- -61-

References to Annex I of D.3. 4. Checklist of phytoplankton

1. Edler, L., Hallfors, G. & Niemi, A. 1984. A preliminary checklist of the phytoplankton of the Baltic Sea. Acta Bot. Fennica 128: 1-26. Acta Bot. Fennica 128: l-26, 1984 2. Haas, L. W. 1982. Improved epifluorescence microscope technique for observing planktonic microorganisms. Ann. Inst. Oceanogr. 58: 261-266.

3. Hoppe, J.E., Daily, R.J. & Jasper, S. 1977. Use of A preliminary check-list of the phytoplankton of the Baltic Sea Nuclepore filters for counting bacteria by fluorescence microscopy. Appl. Environ. LARS EDLER, GUY HALLFORS and AKE NIEMI' Microbiol. 33: 1225-1228.

4. Levine, N.D. et al. 1980. A newly revised classi­ Edler. L., Hiillfors. G. & Ni~mt. A. !984: A prcltmlnary check·l!st oft he phytoplankton of the Balttc Sea.- Acta Bot. Fenntca 128:1-26. Hcls1nk1. ISBN 95l-9469-22-2.1SSN fication or the Protozoa. J. Protozool. 27: 0001-5369 37-58. This check-list summarizes the prest:nt state ol knowledge concerning the spcctes compositwn. nomenclature and, to some degree. the ~ptaxonomy. Protistologica plankton, the nomenclature of the species, and grateful to receive records, corrections and 20: 591-612. to some degree their spatial and temporal improvements in order to make the next edition distributions. By this means we hope to facilitate as useful as possible.

ARRANGEMENT OF THE LIST

For the purpose of the check-list the Baltic Sea Bothnian Bay (BB). the Bothnian Sea (BS), the has been divided into ten subareas (Fig. I), the Archipelago Sea (AS), the Gulf of Finland (GF). the Riga Bay (RB), the Northern Baltic proper 1 Work1ng Group 15 of the Ba1t1c Manne Biologists. (NB), the Central Baltic proper (CB), the Baltic Marine Biologists Publication No ll Southern Baltic proper (SB), the Arkona Basin ..,*'·······-·· ------

-62- -63-

2 L Edla, G. Hdll{or.\ & .4. Nu•mJ ACTA BOT. FENNICA 12H t 191S4J

Explanation of the symbols v ,./···-~~

.,- \KO.:urr.-no: w1thout ccologlCi.!l o.:hur:.~o.:tcnzatlon species belongmg to waters of htgher ~al1n11y than that c 0.:1liJ w<.Ho:r 'PCO.:IC~ 1<10'' Ci of the area ~~o v.urm w:>ter ~pedes 1>10° Cl e matn occurrence 1n eutrophi<:J waters mam oco.:urn:n<:e 1n the ltthHal questiOnable record t're~hwato.:r -.pec1c.:~ whtch Joc.:s not tolerate the t"ull ( ) symbol not very stnct!y

KB AB 58 CB RB GF AS 85 88 CYANOPHYTA (CYANOBACTERIA) Nostocophyceae (Cyanophyceae)

CHRuOCOCC'A LES Aphunon!psu Np<.:r . .-\B =the Ad, una Ba ... l!l. dtHI 1puncww L minor Lagerheim) KB =the Kattegat and Belt Sea Area lll!flU/SSimu Lemmermann) Micmcruo1· P Rtchter sahtdicofa ( L.1gcrheim) Geuler + + ,\.1lcmCI·.I"tn Kut1.1ng anwuno\ll ( Kutnng) Kutz1ng 14) wf wf wf wf wf (AB). and the Kattegat and Belt Sea Area (KB). clatural changes and have included the most (jlu1-aqum• 1 Wtttftl~k l Kirchner) To facilitate rapid location of samples, well important synonyms used tn the Baltic S~!a tn halophtfa \1ancns & Pankow ( 4) defined borders have been drawn which differ brackets below each species. The numbcr(s) 1n retnbofdit (Rlo.:hter) Foni (4) w w w we we + w w somewhat from previously proposed ·natural' brackets after a name refers to an annotation at !Aphanocapw deficali.n·ima W. & G.S.West) ( ,\.-ficron·st1s tncerta { Lemmcrmann) borders. When considering division of the area the end of the list. For the sake of clarity. Lcmmermannl for other purposes the borders suggested by author's names are mostly g1ven m full. For (M. pulverea (WoOO) Foru) Ekman ( 193 I) and by Wattenberg ( 1949) should abbreviations. see Christensen & Thomsen LW. pufverea var. incena tLemmermann) be taken into consideration. (1974) and Parke & Dixon (1976). Crow) wesenbergu ( Species are listed alphabetically under each Ktlm:irek) Scarmach When a spec1es has been recorded from an Rhahdoderma Schm1dle & Lauterborn Order. We have tried to consider recent nomen- area, there is a symbol in the proper column. lmeare Schmidle & Lauterborn- . '·'""~01!:------········

-64- -65-

4 L. Edler. G. Ha'//fors & A. l1ilemt ACTA BOT. FENNICA 128119841 5

KB AB SB CB ~B RB GF AS BS BB KB AB SB CB NB RB GF AS BS BB

NOSTOCALES

Achroonema Skuja ( 5) subtilissima (Ki.itzing) Gamont (8) + /enrum Sk UJa + renuts C.A.Agardh ex Gamont we we proieiforme Skujo + + Phormidium Ki.itzing ex Gamont proteus SkuJO + autumnalr: (C.A.Agardh) Gamont ex Go mom Anabaena Bory e.>; Barnet & Flahault I 11) ba/nca Schmtdt w w w w w w muctco!a Huber-Pestalozzi & Naumann {A. spiroides f. baluca {Schmidt) Pankow) Raphidiopsis Fntsch enema/is Rabenhorst wf medirerranea Skuja ( 37) lA. sptrotdes f. hassallii (Klitzing) Pankow) Sptrulina Turpin ex Gamont cylindnca Lemmermann + + wlf wlf balflca Manens & Pankow lA. subcylmdnca Borge) {6) ma;vr Ki.itz~ng ex Gomor\t inaequalis IKU.tzing) Barnet & wbsalsa Orstedt ex Go mont + Flahauh w w w Th1orhrix Winogradsky ( 14) lrmmermannii P. Richter {7) w w w w + annulara Molisch le {A. flos·aquae f. femmermannu (P.Richter) nl>'ra (Rabenhorst) Winogradsky le Canabaeus) osclllari01des Bory ex Barnet & Flahault r 10) CRYPTOPHYTA scherrmenewi Elenkin (8) Cryptophyceae spiro/des Klebahn (9) wf CRYPTOMONADALES <5) torulosa /Carmichel ex Harvey tn Hooker) Lagerheim ex Barnet et Flahault r IOl Chroomonas Hansgirg ( 18: 19) + vanabi/is Klitzing ex Barnet & Flahault (10, CrrptaultJX sli."uja II. 12) wf manna Throndsen (37) + Anabaenopsis V.Miller Crrptomonas Ehrenberg {3) e/enk1nii V.Miller ( 13) we ba/nca I Karsten) Butcher + + Aphanizomenon Morren ex Barnet & Flahault erosa Ehrenberg + flos-aquae (L.) Ralfs ex Barnet & Ol'ata Ehrenberg + Flahault ( 15) wb w w w w w w w w + pelaf(Jca \Lohmann; Butcher + gracile (lemmermann) Lemmermann w w + + pus1f/a Bachmann + &ggiaroa Trevisan ( 14) Karablepharis Skuja alba (Vaucher) Trevisan wle wle m·alis Sk uja ( J 7) + /epromiuformis ( Meneghinil Leucocryp1os Butcher Trevtsan wle wle manna ( Braarud J Butcher ( II) + mmima Winogradsky wk wle Rhodomonas Karsten mirabilis Cohn wle wle minuw Skuja (I) + + G/oeorrichia J.G.Agardh ex Barnet & Flahauh echinu/ara (J.E.Smtth) R1chter DINOPHYTA Lyngbya C.A.Agardh ex Go mont Dinophyceae conrorra Lemmermann PROROCENTRALES limne1ica Lemme:rmann + + + Prorocenlrum Ehrenberg Nodulana Mertens ex Barnet & Flahau!t btJ//Icum (Lohmnnn) Loeblich 1!! w w w w w w w w harveyana (Thwalles) Thuret ex Barnet & ( Exu\litJ, i/o bailiea Lohmann) Flahault + + + + + cu_\.\uhl(-um 1Wolo~zyr\ska J Dodgl! spumigena Mertens ex Barnet & Flahault ( 15) wb w w w w w w w w + lima /Ehrenberg) Dodge Osci/laroria Vaucher ex Go mont tExuviaella marina Cienkowski) ( 10) agardhii Go mont var. m1cans Ehrenberg w w WS agardhii we we we we mintmum Schiller w w w · var. iso1hrix Skuja wf (Exuviael/a minima P

ACTA BOT FENNI(A 128 I 19X4l 7 6 L. Edler. (i. Hallfors & A. ,Viemt KB AB SB CB NB RB GF AS BS BB KB AB SB CB ~B RB GF AS BS BB

PYROCYSTALES Dissodinium Klebs 1n Pascher + hastata Ste1n pseudolunula Swift + + (+) (W) (w) (W) (W) (w) {W) {w) {W) norveg/Ca C\apun!de &. lachmann !20) (G_~'mnodinium lunula SchUt!) w w w w w w w w w roiUndara C!aparede & Lachmann (Pvronstis /unula (SchUtt) SchUtt) {24) (Phalachroma rorundatum !Clap. & Lachm.) Kofo1d & M1chener} mpos Gourret ( 21) PERIDINIALES Ceranum Schrank arcrwm f Ehrenberg) Cleve + GYMNODINIALES bucephalum (Cleve) Cleve + Actimscus (Ehrenberg) Ehrenberg fUrca f Ehrenberg) Claparede & Lachmann w WS penrasrenas Ehrenberg + tUsu.s rEhrenberg) Dujardin w WS WS ( Gymnaster pentasurias (Ehrenberg) hmmdinel/a (0.F.Mtil!er) Schrank Schtittl homdum (Cleve) Gran w Amplridinium Claparede & Lachmann tmermedium (JOrgensen) JOrgensen + carter/ Hulburt + linea/urn (Ehrenberg) Cleve w crass urn Lohmann\ 22) /ongiper (Batley) Gran w WS longum Lohmann (II. 22) " + + mucroct'fOJ (Ehrenberg) VanhOff~:n w WS (?acurum Lohmann) ./-;;tpo1 tO.F.Mi.illcr} N1tzseh w w WS apercu/alum Clapan!de & Lachmann (8) C/adopyxis Stein pellucidum C. Herdman + + cfavronii R.W.Holm.es + semi/una tum C. Herdman ( 10) Ent::za Lebour'' JWNii ( Lemmerm;~nnl K0fc11d & Swezy + acura { Apsteln) Lebour Cochiodinium Schlitt ( Peridinium /arum Paulsen) helicoides Lebour ( 2:!) Gfenodimum Ehrenberg ( 3) pel/ucidum Lohmann + damcum Paulsen + + folwceum Stetn ( !0) Gymnodinium Ste1n (3, 5) + gracile Bergh + gymnodimum Penard lohmanmi Paulsen + paululum Lmdemann (II) + Hmplex (Lohmann) Kofoid & Swezy iII) + + + + c penard(forme (Lindemann) Schiller ( ll) + a·armingfi Bergh splendens Lebour + Goniodoma Stein westificii Schtitt + 01/enfe/dii Gyrodimum Kofoid & Swezy {5) Paulsen ( l) + aureolum Hulburt w Gonyaulax Dtesing fissum {Levander} Kofo1d & Swezy + + cart•na/0 (Levander) Kofoid cb cb c c c c c c c ( G,rmrwdimum fissum Levander) digiiUI/.1 1 Pouc he{) Kof01d + (SpircxlimumjiSJum (Lev.) Lemm.) nctJvata fBraarudJ Balech + .fusiforme Kofoid & Swezy + uamaren.l'ls Lebour) /onRum (Lohmann) Kofo1d & Swt"zy + .f!.nnt/lcyi Reinecke w w w w w w w w tProtoceratium reticula tum (C!aparede & sp~raie (Bergh) Kofo1d & Swt'Zy + + + Hemtdimum Stein Lachmannl Btit~hhl heft:n.\1\ w~lio~zyriska { 1) nasu1um Stt:1n + ' pol_t•edru Stem ochraceum Levander 123\ + + + rp1m[era fClaparecte & Lachmannl Diesing w w w w w w Kazodinium Fott (5) + tnacantha JOrgensen w w w w w w w (Massartia Conrad) w verior Sournta w w w w w w w rotundatum (Lohmann) Fott + + I diacantha (Meunier) Schiller) Oxyrrhis Dujardin Heterocapsa Stein marina Dujardin (23) + + + -r mquerra (Ehrenberg} Stem w w w w Polykrikos Bi.itschli w w w IPendinium triquetrum (Ehrenberg) Lebour) schwartzi/ BUtsch!i w Micracanthodinium Deflandre Pronoctiluca Fabre·Domerque sel/ferum /Lohmann) Deflandre pe/agica Fabre-Domerque {22) + i C/adopy:ds setifera Lohmann) Torodinium Kofoid & Swezy Oblt:a 8ak.:h ttredo (Pouchetl Kofoid & S~zy + rorunda (LebouT) Balech w w w w we w Warnowia Lindemann w w w {Pendinopsis rotunda Lebour) parva (Lohmann) Lindemann (22) ( Gll?nodinium rotundum (Lebour) Sch1ller) (Peridinium ftmnophilum Lindemann) NOCTILUCALES Pendinium Ehrenberg balticum I Levander) lemmermann + + + + + Nocti/uca Suriray { Glenodinium balticum Levandt"r) scintil/ans (Macartney) Ehrenberg w w -69- -68-

ACTA BOT. FENNICA 128t1984l 9 L. Edler. G. HdlljOrs &. A. Niemt 8 KB AB S8 C8 N8 R8 GF AS 8S 88 RB GF AS BS BB KB A8 S8 C8 N8

p,vn(orme (Paulsen) Balech + (Pmdimum pyr(!Orme Paulsen) cmctum (O.F.Mi.illerl Ehrenberg + wle) + roseum (Paulsen) Balech + hangoei Schiller /Pendinium roseum Paulsen) tgractle Lindemann) ueinit {JOrgensen) Balech + inconspicuum Lemmermann (Pendinium swnii J6rgensen) wtllet Huitfeld·Kaas wbinerme (Paulsen) Loeblich III + Protoperidimum Bergh /Peridimum ~ubinerme Paulsen) w w w ( Peridintum p. p. l + w w Pyrophacus Stein achromattcum (Levander) Balech horo/ogtcum S1e1n w w + IPendimum achromau·cum Levander) + + + Scnpp.1tel/a Balech ex Loebllch III ave !lana Meunier (II l rrochoidea !Stein) loeblich Ill (I I, 25) w w w w w w Peridinium ave/lana (Meunier) Lebour) CW CW IPeridinium 1roch01deum 1 CW CW cw cw CW (Stein) bipts (Paulsen) Balech Lemmermann) {Gienodinium bipes Paulsen) Zygabtkodinium Loebhch tiL & Loeblich Ill (Peridinium minusculum Pavi\lard) !enllculatum Loeb\ich fil. & Loeblich III (26) + + w w w w (Minuscula bipe.r (Paulsen) Lebour) (Diplopsalfs !enucula f. minor Paulsen) brtvt (Paulsen) Balech + ' cw cw cw CW cw cw CW ( Glenodimum /emicu/a f. minor (Paulsen) brevipts (Paulsen) Balech Pavd\ardl (Peridimum brevipes Paulsen) + ( Dtplopel10psis minor ( Pauhcn l Pavi\lard) arasus (Paulsen) Ba!ech Amphtdtmopsts Wo\o~zyris~a (Ptridinium cerasus Paulsen) kotutJ/1 W~!oszyr.!.ka + + daudicans (Paulsen) Balech + "- (Pendinium claudicans Paulsen) + EBRIALES 114) conico1des (Paulsen) Balech (Peridimum conicoides Paulsen) Ebna Bogen + comcum (Gran) Balech mpar111a (Schumann) Lemmermann CW cw cw cw cw c~ CW cw cw (Peridinium conicum Gran) crassipes 1Kofoid) Ba!ech , + (Pendinium crassipes Kofo1d) PRYMNESIOPHYTA (HAPTOPHYTA) + curvipn {Ostenfeld) Ba\ech , Prymnesiophyceae (Haptophyceae) (Pmdinium curvipes Osten\eldl ISOCHRYSIDAI.ES dtclpiens (JOrgensen) Parke & Dodge + (Peridinium decip1ens JOrgensen) + Gephyrocapsa Kamptner deficiens (Meunier) Balech (8) " huxleyi (Lohmann) Reinhardt + + (Cocco/irhus hux/eyi (Lohmann) Kamptner) (Pertdimum defiCiens Meumer) + depressum (Bailey) Balech . + (Emtfiana huxle.vi (Lohmann) Hay & Mohler) (Peridinium depressum Badey) + divergens (Ehrenberg) Balech + Pleurochrysis E. G. Pringsheim !Peridinium divergens Ehrenberg) canerae (8raarud & Fagerland) excenrricum (Paulsen) Ba\ech + T. Christ.:nsen + + (Peridimum excenmcum Paulsen) ( Cncosphaera carraae ( Braanxi & c c ' granii (Ostenfeldl Bakch Fagerlandl Braarud) (Peridimum granii Ostenfeld) ( Hytn~''"/'.·monas carrerae ( Braarud & tPeridinium finlandicum Paulsen) . Fagerlandl Manton & Peterfll {Peridimum divergens var. levanden PAPPOSPHAERALES Lemmermann) leoms (Pavi\lard) Balech + Acanrhoica Lohmann (Peridinium leonis Pavillard) quamospina Lohmann + longispinum (Kofoid) Ba\ech (22) Balaniger Thomsen & Oates oblongum (Aurivillius) Parke_e~ Dodge + balticus Thomsen & Oates + (Peridinium oblongum Aunvdhus) Braarudosphaera Deflandre oceanicum (VanhOffenl Balech btgelowii (Gran & Braarud) Deflandre + (Peridimum oceanicum VanhOffen) Ca(rptr01phaera Lohmann + ovatum Pouchet + Di1·cosphaera Haeckel (Ptridinium ova tum (Pouchet) SchUtt) tub(fer (Murray et Blackman) Ostenfeld (8) ? CW pallidum Ostenfeld) Balech Pappomonas Manton & Oates (Ptridimum pa/lidum Ostenfeld) w w + vtrgulosa Manton & Sutherland CW + w + pellucidum Bergh . .. (Pmdinium pef!uCidum (Bergh) Schutt) PRYMNESIALES penragonum (Gran) Balech + Chrysochromu/in.a Lackey ( 5) (Per~dinium penragonum Gran) acanrha Leadbeater & Manton + punctulatum {Paulsen) Balech + (Pmdinium puncrulatum Paulsen) -71- -70-

ACTA BOT. FENNICA 12811984> II

L Edll'f. (i. Hdll{vrs & A . .t•b!'mt KB AB SB CB 1'18 RB GF AS BS BB 10 BS 88 NB RB GF AS KB AB SB CB bavancum Imhof cylindncum Imhof Mant;!roh!lu.\ P:.~rkc ..:\ Yl..HI\Iln 1n Park..:. Manton & uve/la Ehrenberg 129) + r + Croglota Ehrenb..:rg (Iarke + prramtdosa Thomsen amencana C:.~!kins w w w w 1·o!vox Ehrenberg PhaeocySIIS Lagerhetm + amoeb01dea BUttner (22) pouchew ( Harriot) Lagerhe1m CHR0MULINALES T sphaeroides BUttner ( ::!2) Apedmel!a Throndsen Prymneswm Massan + + + 1P1mfaa !Tilrondsenl Throndsen + + parvum Caner (II) Calw omnnas Lohmann ( 11) Tngonaspts Thomsen + f!\•alo- Wulff w w w d1skoens1s Thomsen + vungoon !C<>nrad) Lund w w w m111Utiss1ma Thomsen wu/tfn' Conmd & Kufferath w w w w w W1gwamma M:.~nwn. Sutherland & 0ates + Chr\·j·ococcus Klebs scenozonwn Thomsen bt,70rus Sk u)a ru,·ercen~· Kkbs PAVLOV ALES Keph1rion Pascher hemuphaericum ( Ladcv) Conrad + Pavlova Butcher !5) + Pcdinella Wy<,otzki · /when

ACTA BOT. FENNICA 128 119841 13 12 L. Edler. G. Hdllfon & A. N1emt KB AB SB CB NB RB GF AS BS BB AS BS BB KB AB SB CB NB RB GF

lens Ehrenbo:rg _._ Tribophvceae (Xanthophyceae, Heterocontae) mobi/JenHs (Bailey) Grunow ex Van Heurck + RHIWCHLORIDALES obrusa !Kutz!ng) Ra!fs in Pritchard + pulfhella Gray (4) + + _._ Rh1:och/orrs Paschcr rhombus (Ehrenberg) W. Smith + nodufanae Bursa + 5/nen.m Greville + so/irana Bursa )uhaequa ( KUtzing) Ralfs in Pritchard + Cera/au/ina Peragallo MISCHOCOCCALES pela.~!Ca (Cleve) Hendey w tba.~onil Peragat!o) .Weringosphaera Lohmann (I. 141 Ceraraulus Ehrenberg mediterranea Lohmann + + 1urg1dus (Ehrenberg) Ehrenberg rad1ans Lohmann + + Chaetoceros Ehrenberg serrara Lohmann a(finJJ Lauder {4~) Ophwcynum Nageli CW r anastomosans Grunow e/ongatum W & G. S. West + at/anucus Cleve Schil/emlla Pascher s + borealis Bailey anuraea Pascher (!. 30\ + T bre\1/s SchOtt + + ca/curaf!S !Paulsen) Taka no + TRIBONEMATALES !Hmplex var. calcitrans Paulsen) (I l) Tnbonema Derbes & Solier If cerai05porus 0stenfdd ( ~2) cw CW cw CW c(w) C(W) c(w) (w) affine G. S. West If nnctus Gran T v.iride Pascher comprenus ·Lauder CW + < oncal'lcorne Mangin <'onsrrictus Gran + ,. emrnltnus Castracane f. com·olun.~s LES) + Diatomophyceae (Bacillariophyceae) - r mseto5a Brunei EUPODISCALES iBIDDULPH!ALES. CENTRA c•nonarus Gran T' coHutus Pavdlard c Acanrhoceras Honigmann mnuus SchUtt ( 33) IC)W ;achanasii ( Brun l Simonsen cun·1serus Cleve {Ailheya ;achariasii Bruni CW + damcus Cleve ( 34) cw cw cw CW Ac 11 nocyclus Ehrenberg {C)W (c)w + (c)w (C)W deh1/i> Cleve cw T octonanus Ehrenberg denptens Cleve (ehrenbergii Ralfs) w w + + cw CW w w w + denws Cleve _._ _ var. ncronarius + + dwdema (Ehrenberg) Gran c(w} _ var. crassus (W. Smnh) Hendey I 10) T !.1Ub1·ecundus (Grunow) HustedU _ var. 1enellus 1Breb.) Hendey I 10\ dHhmus Ehrenber~ cw A.cllnoptychus Ehrenberg gra< i11s SchUtt senanus {Ehrenberg) Ehrenberg + + ho/.wticus SchUtt (undulatus (Bailey) Ralfs) be c c c mgo(lianus Ostenfeld in Gran c Auhe1·a T. West ' lunmosus SchUtt c + de~ora T. West .'uudai Ralfs !n Lauder + A.ulacosira Thwa1te:-; furenzl;ua l Grunow\ Stmon:>en W< WO WO WO WO rerpusdlus Cleve + dista~s (Ehrenberg) Stmon~n (31 l preudocrinuus Ostenfeld l(ranulata (Ehrenberg) Stmons.en + rod1ans SchOtt T '_ var. f(ranulata . radicans SchUll - var. angustissima (0_ MUller) Simonsen + 'it'lrOCan!hus Gran w w isfandica (0. MUller) Simonsen \·cp1enmonalis bstrup cw CW ssp. hetvn1ca {0. MUller) Simonsen cw CW CW cw CW r1mt/is Cleve c ~ italica (Ehrenberg} Simonsen + f Simplex Ostenfe!d + + ssp. 1talica var. ltalica . f wC!alis Lauder + - var. va/ida (Grunow) S1monsen f subtilis Cleve c(w)' ssp. subarctica (0. MUller) Simonsen + + + teres Cleve c + ' Bacreriastrum Shadboh .-.cighamii Brightwell {33) CW CW cw cw CW CW elongarum Cleve + _._ cw cw cw + 1\'tliel Gran hyalinum Lauder Corethron Castracane Bacteriosira Gran + cnophi/um Castracane .,. fragllis Gran Coscmndhcus Ehrenberg Biddulphia Gray antra/is Ehrenberg + aurita {Lyngbye) Breb1sson + + granulata Roper +

6 4831B4A -75- -74-

ACTA BOT FENNICA 128 (19841 15

L. Edler. (J_ Hdll(vrs & A ."•hem/ 14 KB AB SB CB NB RB GF AS BS 88 GF AS BS 88 KB AB SB CB NB RB

lon_((tJl'IO Zachanas + + mtntma Le\·ander w we concmnus W. Smith w + w + + + + w + pun_!ffrtJ A. Cleve + granii Gough (35) H.'lf.f?t'ra Brightwell + + granulosus Grunow ( 10) + ~-hruhYolei Cleve f ocuius·irld/5 Ehrenberg (36) + + Holtedothn Peragal!o + radiarus Ehrenberg Hyliform1.'i Brightwell + rorhii {Ehrenberg) Grunow + Ropena Grunow I!;( Van Heurck sub-bulliens Jorgensen /eHellara ( R1)per\ Grunow Cyclore!la Ki.itzing we Skeletonema Grevllle i52) atom us Husted! { 37) w w w w .,_ w COI'/Otum tGrc=vdkl Cleve cw cw cw cw {C)W c(w) T C(W) C(W) caspta Grunow wbsalsum I Cleve· Euler) Bethge e e comta !Ehrenberg) Ki.itzing + Stephanodiscus Ehrenberg kuelttngtana Thwaites (3~) f a:straea I Ehrenb~rgJ Grunow + meneghimana KUwng r (asrraea var. mmutula (Kiltzing) H~ndev) srelligua Cleve & Grunow ~ (")row/a I Kiltllng) Hendev) · smora (Kilwngl Grunow in Cleve & Grunow + + dubws I Fmkel Hustedt - Deronula SchUll c c hant:schli Grunow (..1.!) confervacea (Cleve) Gran lhant:.1chtt var. pu.1tlla Grunow) pumi/a (Castracane) Schlitt + !Schroederel/a delicawla (Peragallol StephanopyHI Ehrenberg turn~ ( Grevllkl Ralls m _Pntchard Pavdlardl .'J"trepwrht'Cil Shrub~ol~ Dtll·lum W. Bailey bnghtwellii (T. West) Grunow ex Van Heurck w tame.\'/.\ Shri.{b<.o!e + Thala.ISIOI"IrO Ckve Eucampta Ehrenberg + ant;?UJte-lmeata (A. S<.:hm!dtl G. Fryxell :odiacus Ehrenberg Vwnardia Peragallo & Hask c flacdda (C:lstracane) Peragallo w ICtHCinoilra pol_1chorda !Gran) Gran) haluta IGrunnw) O'>tenfeiJ + + + O::(W) <;(wl + C(W) C(W) Hyalodiscus Ehrenberg + scu11cus ( KUmng) Grunow + bramaputrae I Ehrenberg) Hf.tka n~~on -i- + Lauderia Cleve & Lo<.:kl.'r + c T I Thalasswsira lacu.l'lrH 1Grunow) Haslel ' borealis Gran (annulata Cleve) rCoscin()(ltSCIH lacu.l/rl\ Grunow) dectpten.Y !Grunow) Jnrgenscn Ltptocy!indrus Cleve + eccenmca 1 Ehrenberg) Cleve +' ' damcus Cleve + mediterraneus (Peragailo) Hasle tCoscinoducus acentncus Ehrenberg) gwvida Cleve c tDacty/iosolen mediterraneus Peragallo) + + + ~ m1mmus Gran (39) <.:utllardii Hasle -i- Ltthodesmium Ehrenberg h_~alina (Grunow) Gran + f undulatum Ehrenberg .'eptopus !Grunow) Hasle & G. Frvxell I lmeaws Melosira C. A. Agardh c ~O.\CtnodtS<"US Ehrenbe~gJ c c c arcttca (Ehrenberg) Dickie ' ' ' !t·~·amlen van Gonr norden.~J.:toeldn Cleve ' dubia Kiitzing lineata (Dillwyn) C. A. Agardh pseudonanu Has!e & HeJmdal w Uuergensii C. A. Ag.ardh) rotula i~lcun1cr + moniiiformis (().F. MUller) C. A. Agardh !{"eJn:{logti /Grunow) G. Fryxell & Hasle nummu/oides (Dillwyn) C. A. Agardh tj1uwatili.1· Husu:dtl varians C. A. Agardh BACILLARIALES iPENNALESl Paralia Heiberg Achnanthes Bury (42) sulcata (Ehrenberg) Cleve hw.wlettiana ( K Utzing) Grunow Podosira Ehrenberg hrt'l'tpes C. A. Agardh montagnei KUtzing (22) delicatula IKUtzing) Grunow em. stel/igera (Bailey) Mann Lange-Bertalot Porosira JOrgensen in Nordgaard - var. delicaiU/a glacio/is (Grunow) JOrgensen c gibberula Grunow Rhi:oso/enia Brightwell + + I hw ~oletllana ( KUtzmg) Grunow alara Brightwell + Jn Cleve & Grunow) calcar-avis Schultze + taentata Grunow c c + c cylindrus Cleve ( 40) + Amphtprora Ehrenberg ' ' ' de/icatula Cleve .a law Ki..itzi ng ( 1 1J eriensis H. L. Smith w + (+) (+) kjellmanntl Cleve c fragilissima Berg + orna1a Bailey hebetata f. semi:spina (Hensen) Gran ... imbricata Brightwell -77- -76-

ACTA BOT. FENNICA 128119841 17

L. Edler. 0. Hdllfon & A. N11!m1 16 KB AB SB CB NB RB GF AS BS DB GF AS BS BB KB AB SB CB NB RB

ventralls t Ehrenberg) Hustcdt Fragtlarw Lyngbye {42) pa!udo>a W Sm\\h var. pa!udo~a capucma Desm;:LZ!eres - var subsalina Ckve + consrruens (Ehrenberg) Grunow var. Amphora Ehrenberg {421 construens If coffeaeform!S IC A. Agard hi Ki.iwng + + + - var. rub salina Hustedt If - var. cojfeaeform/S + + cro10nensu Kitton f - v n Cymbe/la Agardh - var. rent'fa ( KUtzing) Cleve I aspera (Ehrenberg) Cleve n dm•eta { W Smith) Ra!fs 10 Pnt..:hard emu/a (Hempnch) Grunow 11 ('/t·guns W. Sm1th ventricosa KUtztng + gr,•(!ana Donk1n ill + + Diatoma De Candolle (+I (+I i+l (+I ill humaosa Breb1sson elongarum (Lyngbyel C. A. Agardh I I I manna Ra!fs vulgare Bory var. constricta Van Heurck pela.(!tca Cleve ( 10) Dip/antis Ehrenberg {42) + pere.~nna (Ehrenberg) KUtzing var. paegr~na didyma (Ehrenberg) Cleve - var. kefvingiensts (Ehrenberg) Cleve smithii (Brebisson) Cleve pseudomemhranacea Cleve-Eu!er Epithemia Brebisson {42\ PY.~maea Ki.itzing (53) sorex Ki.itzing rad1ow KUtzing If turg1da (Ehrenberg) Ki.itzing rhomhtca Gregory zebra (Ehrenberg) Kiltz1ng rhynchocephala Ki.i1z1ng Eunotia Ehrenberg (42) ptctinalfs {Ki.itzing) Rabenhorst var. -78- -79-

ACTA BOT. FENNICA 128 ( 1984) 19 18 L. Edler. G. Hiillfors & A. Ntemi BB KB AB SB CB NB RB GF AS BS 88 KB AB SB CB ~B RB GF AS BS

- var. venmcosa !Kiitzing) Grunow saltnarum Grunow gibbuula (Ehrenberg) 0. Muller transuans Cle ve var. tluesu (Grunow) Cl eve + Stauroneis Ehrenberg !42) f. de/icatula Hetmdal membranacea (Cleve) Hustedt + trrpunctata (0. F. Muller) Bory spicula Hickte + (gracilis Ehrenberg) c c Sunopterobta Brebisson •·anhneffemi Gran tntumedia L.ewts !43) ••mdula Kii!Zing .._ Surirella Turpin ( 42) ."'II:!Chta Hassall (42 ) If capromi Brebisson 143) If aCicularrs W. Smtth + ~emma (Ehrenberg) Kiitzing + + deltcausstma Cleve or + ovalir Brcbisson var. ova/is ( I) I clnsterrum t Ehrenberg) W. Smith I I. I I) + + + + + c c - var. crumtna (Brcbisson) Van Heurck ( I) I cylindrus !Grunow) Hasle ovata Kiitzing (I) I ( Fragtlarra cyltndrus Grunow) Striatula Turpin ( 43) I (fragtlariopsis c.vltndrus (Grunow) Krieger) tenua Gregory var. nuvosa A. Schmidt (43) If dubta W. Smith Synedra Ehrenberg (42) Jiliformts ( W. Smith) Hustedt acus Klitzing var. acus If I [on/leola Grunow c c c - var. angusrisstma Grunow f + f ngrda Grunow I pulchella Ralfs I I f rustulum ( Kiitzi ng) Grunow I( I) rum~n.f Klitzing If gracilis Hantzsch ex Rabenhorst tah11/ata !C. A. Agard h) Klitzinl( (45) I grunowii Hasle + ulna (Nitzschl Ehrenherg If (fra.~i/arra oaanica Cleve) •·uuchrriae ·Kiitll ng If (fragi/ariopsis oaanica (Cleve) Hasle) Tuhrllurra Ehrenberg hungarica Grunow ( 43) f entnrura !Lyngbye) Kiitztng If I hybrtda Grunow (43) ,11occulosa !Roth) Kiitzing If If mtermedia Hantzsch ex Cleve & Grunow Tha la.uwnema Grunow ex Hustedt (capitella Hustedt) + .._ nu:schiotdes Hustedt c longtSSima I Brebissonl Ralfs + + + + + I Thulasstothnx Cleve & Grunow lorenztana Grunow (43) I (rauenfe/dii Grunow in Cleve & Grunow + mtcrocephala Grunow I longtwma Cleve & Grunow + paleo !Kiitzing) W. Smith (l)w Ill Tropidoneis Cleve pa/eacea Grunow (44) w w "' "' I dann/tltii Cleve-Euler ... + pupustl/a (Kiitzing) Lange-Bertalot lt ptdopttra (Gregory) Cleve + pseudodeltcuttssima Hasle + puncrata ( W. Smith) Grunow (43) Raphidophyceae ( Chloromonadophyceae) p ungens Grunow + If pu;tl/a !Kiilllng) Gruno w em. Lange-Bertalot RAPHID0MONADALES tkutt:rngiana Hilse) fionyosromum Diesing ( 18) sca/aris (Ehrenberg) W Smith (4.1 ) sutata Cleve c ~ - f. ohtusa Hasle + EUGLENOPHYT A stgma !Kiilllng) W. Smith (43) Euglenoph) ceae stgmoidea !Nitz~hl W Smith (4Jl subtilis ( Kiitzing) Gruno w H JG LE NALES tryblionella Hantzsc h (43) Afla.Ha 0Ujardin (5) + w!rmicularis (Kiitzing) Grunow (431 t ula<'lum F.h re nberg Optphora Petit arbu;-cu/a Stetn marryi Hcnbaud venculo.rum Ehrenberg Phatodactylum Bohlin + Eu((lena Ehrenberg (42) tricornurum Bohlin (23) !I CuJ Ehrenberg f Pinnularia Ehrenberg a .~.1·uns Schmarda quadratarea (A. Schmtdt) Cleve var. c c c c proxima Dangeard + stuxbergii C:leve (I) rplfngyra Ehrenberg f Pleurosigma W. Smith (43) I mpttm ( Dujardin) Klebs f angulatum !Quekett) W. Smith I vtridis (0. F. Muller) Ehrenberg ef elongarum W. Smith If Eutrepria Perty ( S) + subsalsum Wislouch & Kolbe lanuwii Stuer + + Rhotcosphenia Grunow virtdis Perty + abbreviara !C. A. Agard h) Lange-Bertalot Eurrepuel/a da Cunha (5) + + + (curvata (Kiitzing) Grunow ex Rabenhorst) braarudii Throndsen + Rhopalodia 0 . Muller gymnastica Throndsen + + grbba (Ehrenberg) 0. Muller va r. gibba -80- -81-

ACTA BOT. FENNICA 128 ( 1984) 21 20 L. Edlu. G. Hdllfors & A. Nt~mi BB KB AB SB CB NB RB GF AS BS 88 KB AB SB CB NB RB GF AS BS

Chlorophyceae L~pocinclt s Perty ( 5) VOL VOC ALES (incl. TETRASPORALES) ovum Ehr~nberg em. Lemmermann Phacus DuJardin ( 5) Brachcomonas Bohlin · fongteauda (Ehrenberg) DuJardin rubmarina Bohlin (23) + pl~u ronecus ((). F. Muller) Du jardin Ccm~ria Diesmg (5. 54) + + + + + pyrum ( Ehrenb~rg) Stein Chlamydomonas Ehrenberg (5) + + + Strombomonas Denandrc marina Cohn (I) c d~J7andm (Roll\ Der1andre Eudonna Ehrenberg Trachelnmonas Ehrenberg elt'gans Ehrenberg htsptda (Pertyl Stem em. Denandre unicocca G. M. Smith volvoccna Ehrenberg Glo~ocysm Nageli planctomca 1W . & G. S. West) Lemmermann Conium (). F Muller ptCiora/~ 0 . F. Muller CHLOROPHYTA roc cale ( DuJa rdin) Warming Prasinophyceae (incl. Loxophyceae) Pandonna Bory em. Ehrenberg PEDINOMONADALES murum (0. F. Mlillerl Bory + Pla nktococcus Korsikov Pedinomonas Korsikov (5) 1pha~rocyst: fo rmis Korsikov PTEROSPERMATALES Planktnipha~na G. :vi. Sl)'lith ge/a11nosa p . M. Smith Micromonas Manton & Parke + + + ... Sphaeroo-stis Chodat em . Korsikov pustlla (Butcher) Manton & Parke Rhrn~tui Chodat + + Puudoscourfie/dia Manton -t- marina (Throndsen) Manton CHLOROCOCCALES Mantoniel/a Desikachary + Ac11nastrum Lagerheim squamata (Manton et Parke) Desikachary + ha111:schit Lagerherm Nephroselmis Stein ? Anktstrodesmus Corda minuta (Carter) Butcher (a/cat us (Corda) Ralfs (I) olivacea Stein em. Moestrup & Ettl ~ Botryococcus Kiitzing pyriformts (Carter) Ettl braunu Kiitzlng + + + + rotunda (Carter) Fott Cuelastrum Nageli Pterosperma Poucher + + cambricum Archer f crista tum J. S.:hiller mtcroporum Nageli f vanhoefftnti (Jorgensen) Ostenfeld + rwculatum 1Dangeard) S~nn f in 0stenfeld et Schmrdt + Chuduul/a Lemmermann em. Fott Pachysphaua {)stenfeld in Knudsen et Ostenfeld (ubsalsa Lcm mermann pe/agica Ostenfeld in Knudsen et Ostenfeld + Cruc t ~~nlcJ Morren quadrata Morren e MAMIELLALES 'tCtangularis !A . Braun! Gdy "'- Mamtella Moestrup l~trap~diu !Kcn:hnerl W. G. S. West gilva (Parke & Rayos) Moestrup (49) + Dtctyv.< hui (Lemmermann) G. M. Sm11h Pyramimonas Schmarda (5) Col~nl<~nta Chodat em. Korsikov grossi Parke + radJala Chodat obovata N. Carter + + Kirchner i~/la Schmrdle + + + orienta/is Butcher + contona (Schmidle) Bohlin e PRASINOCLADALES lunam (Kirchner) Mobius obesa ( W. West) Schmidle Tetraselmis Stein (55) Lugerheimia Chodat canvolutat (Parke et Manton) Norris, Hori xen~vens1s Chodat & Chihara + + longtseta (Lagerheim) Printz cordiformis (Carter) Stein Micractinium Fresenius (Cartena cordiformis (Carter) Diehl) pustllum Fresenius (Piarymonas tetrathelt G. S. West) -83- -82-

ACTA BOT. FENNICA 128 ( 19841 23 22 L. Edltr. G. Hiillfors & .f Ni~mt KB AB SB CB NB RB GF AS AS BS BB BS BB KB AB SB CB NB RB GF - var. Jisetformts Chodat f ~ tllipsotdeus Chodat f Monoraphidium Koniarkova·Legnerova granulatus W. & G. S. West braunu tNagelil Komarkova -Legnerova + gutwinskti Chodat f (Ankwrodtsmus braunii (Nagelil intermedius Chodat + + Brunnthaler) le!(vrti Deflandre f capricornutum (Printz) Nygaard obftquuJ (Turpin) Kli!Zing tSeltnastrum capricornutum Printz) + + tacutus Meyenl contnrtum (Thuretl Komarkova-Legnerova -t- + + + + + opolitn.riJ P. Richter + + (45) o•·alternu.~ Chodat f (.~nkwrodtsmus angustus Bernard ?) (t) .,. wnt Hortohagyi f grtffithu ( Be rkeley) Kom:lrk ova-Legnerova 1ptnost11 Chodat t.-lnkiSirodesmus acicularis (A. Braun) + Schroedtrta Lemmermann Korstkov) + + stll ~ua IS.:hrnderl Lemmermann minutum 1 Nagelil Komarkova-Legnerova I Ankistrodesmus settguus (Schroder) (Stltnasrrum mtnutum tNageli) Collins) + + + G. S. Wesn mtrabtlt tW. & G. S. West) Pankow Sorastrum Klitz1ng (Ankisrrodesmus mtrabilis I W. & G . S. sptnulo.•um Nageli West) Lemmermannl + Tnraedron Klitzing se11[ormt (Nygaard) Komarkova-Legnerova coudatum !Corda) Hansg1rg (Arrkistrodesmusfalcatus va r. setiformis limnettcum Borge Nygaard) mtmmum (A. Braun) Hansgirg + Nephrocytium Nageli f plane tomcum G. M. Sm1th f agardhianum Nageli f quadratum -~ Remsclil Hansg 1rg lunatum W. West regulare Kiitzmg var. ngulart 14) Oocystis + ... 1- Nageli 13) + + + + + + - var. tncus Teiling (4) borgti Snow + + .... + + + trt~onum (Niigeli) Hansg~rg lacustrts Chodat Treul>arta Bernard parva W. & G. S. Sm1th + + + trtuppendtculata Bernard pelagica Lemmermann + + Trochtscw Kii!Zing solitario Wittrock 1n Wittrock et Nordstedt + + + + + + + brachtolata (Mobius) Lemmermann + submarino Lagerhe1m + Pediastrum Meyen arrgulosum (Ehrenberg) Meneghmi ULCJTR ICH:\ LES biradiatum Meyen bor,vanum (Turpin) Meneghim r. Elakatothrtx Wille clathratum (Schroterl Lemmermann f !!tlattrrosa Wille 1 II) \'trtdtJ (Snow) Printz (II) consmctum Hassan Klebsormtdtum Silva. Mauox & Blackwell duplex Meyen r. (graci/limum (W. & G. S. West) Thunmark) j lacndum I Klitzing) Silva. Mauox & tlimneltcum Thunmarkl Blackwe ll II) c f Ko ltel/a Hindak integrum Nageli f ion~i>tta Hi ndak f. tenutS Nygaard (37) kawraiskyi Schmidle + f f'lanktontma S.:hm1dle muticum Kiitzing + prtvum (Printz) Hegewald lauttrbor'lt! s~t-:nldle + + + + + + + I Btnll<'!l'arta lauttrhornit 1Schm1dle) simplex Meyen Prosh kma-Lavrenkol tetras (Ehrenberg) Ralfs Polyedriopsis Schmidle spinulosa Sc hmidle O!:DOGONIALES Scerrtdesmus Meyen (42) + + + + acumirratus (lagerheim) Chodat f Oedogomum Link ex Hirn (46) apiculatus ( W. & G . S. West ) Chodat arcuatus Lemmermann - var. arcuatus ZYGNEMATALES - var. platydiscus G. M. Smith + armatus (Chodat) G. M. Smith Arthrodesmur Ehrenberg f tncur 1Brebls >on) Hassall btcaudatus ( Hansgi rg) Chodat + + Clo.rttrium Nitzs~ h (42. 47) bicellulam Chodat (II) f unculare T. West + brastlitrrsu Bohlin f O<'Utum Brebisson var. voriabtlt brevisptrra (G. M. Smith) Chodat I Lemmermann) Kneger carinatus (Lemmermann) Chodat + + montltfaum ( Bory) Ralfs communis Hegewald Cosmarium Corda (47) + (quadricauda auct. p. p.) + + Mou~rotia C. A. A!!ardh (46) I derrticulatus Lagerheim f ecornis (Ralfs) Chodat var. ecornis -84- -85-

24 L. Edln. G. Hiill(ors & A.. Vtrmt ACTA BOT. FENN ICA 128 ( 1984) 25 KB AB SB CB :"liB RB GF AS BS BB 140) R. rrltndrus ts an a !ten 1n the flora nf the Balttc Sea . The o.:C,IS ional re.:ord by N1em1 & Hallfors ( IQ74) p robablv ( 48) The C. cf. afjims of Niemi & Hallfo rs ( 1974) ts probably Spirogyra Link (46) stems from ballast water. · an undescnbed spectes lcf. Hallfo rs 1979) ( ~Q) Moestrup I 1984) · Staurustrum Me ye n (42. 47) (41 ) S . han1:.1chu va r. pusilla has re peatedly been reported gracile Ralfs ex Ralfs from eutrophted waters. In Finnish literature. however. (50) The r~co r d from GF of Luther ( 1934) of Hym~nomonas roseola Ste1n . appears to belong here. planktomcum Tctl ing th., epit het probably covers several 5mall ce ntric Zl'gnema C. A. Agard h (46) spectes. among others Thalasstostra J?uillardii. T. ( 511 For a discussion <>f the species co ncept 1n the genus. see Z1mmermann et al. (in press). pseudonana and Cyclotella atom us ( Hallfors & Hall fors 1983). (52) In add ition. Kell ( 198 1) gi\'es S. pot amos (Weber) Hasle for the eastern and central Ba lric Sea. (42) Additional speetes of this genus occur occasionallv in (53) Kell ( 1981) the plankton. · NOTES (4)) Matn occurrence 1n shallow ha\'< •n the innermost (54) Mos t records appear to be of m t ~ lden t 1 fied archtpelago. Pyramimonas and Tn raselmts ~ pe ci es . f 55\ Norris et al. ( 1980) (44) In Edler ( 1975) and Hiillfors 1 1979) t h 1~ spedes is called (I) Requtres taxonomtc re -1 nvesttgation. ( 27) T he record ts based on the ob,ervation of s1ngle >Cale' N acunastrotdes. ( 2) Hal me & Molder ( 1958') further report the ~pectes C. onl y. 145) Accordtng to Patnck & Re1mer 1 1966) the cNrect name ACKNOWLEDGEMENTS dubtum Grunow and C. minuussimun Lemme rmann 12l!l Martem & Pa nkow 11972) and Pankow 11916) 1n would be S. fasctculata IAgardhl Kiit ztng. from the PoJO Bay. See. however. Pankow ( 1976· 261. add1t10n re.:ord Btcoeca orata Lemmermann. wh1ch. 146) Sten le. accidenticall y planktonte lilaments can ( 3) All species tn th is genus need taxono mtc re­ however. probablv does not belong here . generally not be determtned to the spec1es . We thank Tvtirminne · > t>!o gi~al Station for good working In vestigation. ( 29) E~pecta lly in older ltter:lture the spectes has been (47) Undetermined $peetes •>f •hts ge nus oc.: ur mt> re or les< factltttes and the Nattonal Swedish Environment Protection (4) See Pa nkow ( 1976). comprehended 1n a collective sense. including the other regularly in estuarine areas Board fo r covering travel expenses. D r. G. Russell ILtve rpool) k10dl y checked th e English. (5) In all areas probably ~eve ra l spec1es are present whtch spec1es of the 11enus ( = S. uvt'lla Ehrenberg) Such need to be worked out . determinations ~hould rather be gtven a~ Synura 'P· (6) Recorded as an Independent species in Hallfors ( 1979). Sensu stncto the spectes should be Cited S. avella Ste1n ( 7) Thts spectes has mostly been recorded as Anabaena flos­ em. KorstkO\'. Electron micros.:opy of the scale' 1s aquae (Lyngbye) Breb1sson. In our opinton A. flos­ usually requtred for correct d eterm10auon of most aquae and A. le mmermannit should be kept as separate spec1es of the genus. spectes. All samples With akinetes from the BaltiC Sea (30) Recorded by :-.: 1em1 et al. ( 1970). Sys tema tiC pO\Itlon REFERENCES that have been stud1ed have been A. lemmumannit. It uncert:lln. see P~rke & D1xon ( 1976: 557). The Ba lttc has also been confused with A. baluca (Niemi & Sea fi nd 1S probably a reduced iorm of Chaetoceros B ii tt n~ r . J. 1911 D1e farh1gen Flagelhten de' l<1eler Hafens & Hallfors 1974 ). wbtilis. - Wt<~ . Mee r e~un te r ; .. At-t Kiel. ':. F. 12:119-133 Ha l m ~. E. \f o ld~r. K JOS S: Plank ·. 'ogtsche (8) Given by Pankow ( 1976) for the southern Balttc Sea. ( 31) In addition. Hal me & ~! olde r ( 1958) me ntton f. sutatu Chm tensen. T. & Thomsen. H. A. 1914 .\lgd'ortegnels• L'n tersuo: hungen 10 cier Poir>-Bucht und angn:nzenden (91 Not 1n the sense of Pankow ( 1965). 0 . Muller. var. alptgena Grunow and var. lt rata -}5 pp .. Kebenhavn. .. ?cwassern. Ill PhytoplanktN>. - Ann. Bot. Soc. Va namo' 30())· 1-7 1. ( 10) Given by Pankow ( 1976) for the Balt1c Sea. 1Ehrenberg) Bethge from the tnnermost part of the Po1o Droop. M. R. 1953: On the ~c o l ogy o f nagell a te~ from some (II) The determinatton needs conftrmatton. Bay bracktsh and fresh water rockpool< of F1nla nd. - Acta Hensen. V . 188 7 !''ber d1e Bestim mung des Planktons. - ( 121 Record ed from th e sea only without ak1netes. Akinete (32) Leegaard 119201 Incorrectly u'ed the name C. debt!H fo r Bot. Fenntea 51 : 1-~2. 13er. Komm . Wt~ s . Unters. deu• ~ he r Meere in Kiel 12- 1~ : 1-108. fo rma t1 on 1s at t1mes abundant 1n ce rtain brackish­ th1s ~ pe ct e s . Edler. L. .197;. Seasonal changes of phytoplank ton 1n the water rock-pools ly heen !•·hland. bs(erreich und Sy mpo~1 um on Manne B1 ology. Gdansk 1975. ) I) pp. 1 ( 13) Jarnefe lt 11964) re ports what is probably th 1s species determ1ned as C cnnuus accord tng to Hu,tedt I IQ27- 1m1 mcogr. ). der S~ tweu; '1 I ).1 -Q~O larne1e.lt. H. (Q64: trber das K.1mmerplankton e1ner under the name Anabaenopsts ctrculan s. 19301. However. gen u1ne C. cnnuus •~ quite J1ffercnt Ekman. S. 1931 : Vorschlag zu e:ner naturwls~en t the lh•ra tn th e ( 34) In add1ttvn to >OIItJf\ cd!s the spectcs also form' .:halO> Int. Revue ges . H ~ d r o h 1 ~ l. 25. 1(>1 . 1 ·' · ·~ .f :9t '· -Acta Soc. Sct ent. Pseudunabaena. {!>~\er:.! p... per)J. - ~ ed • ' . Darma rk~ Fiskert- og Ha v under~ . N.S ~f1 y Fen n•cac 41!( 5) . 1-~4 ( 17) Acctdentally planktonic filaments are eastly confused ( 35) N1c m1 et at. 11970) report thts sp.:ctes erroneously as C. If 12\: 1-J .l. Luther. A. 1934· Uber e1ne Cc• cc.-,:.:~ ilee a u ~ dem with Oscillatona spp. oculus-trtdts Ehrenberg. Apparently the same spe.:tes 1s ~! all f ors. G. 1979: <\ preltminar: ched ·ltst of the Ftnn1schen Meerbusen. - Memoranda Soc. Fauna ( 18) The spe.:ies have not been worked ..; ut. also reported as C. asteromphalus Ehrenberg tn several phytoplankton of th e northe rn Balu.: "··~ - Pubis. ( 19) C. balttca ( BUttner) Carter and C marina ( BUttner) papers. C. grami IS well .:hara.~ te rized by the eccentrt.:al Water Res. lnst. 34·3-24 Flora F·: nn1ca 9·16~ ·I 71. Butcher recorded from the shore at K1el (BUttner 1911 ). cu rvature of the •ahe. g1rdle bands tapenng to wards Martens. B. & Panh•w H. 1972: Taxonomtsche ~ ;ill fo ~ s. G. 1984: Ftlamentous rock-p

5. Rubin codes of phytoplankton 26 L. EJ/er. G. Hiilljim & A. Niemi algae- thtrd revision. - J . ma r . btOI. Ass. U. K. 5(>:521- Ialt. Julk./Havsf•.H'I.nln!!Sinst Skr 2.U: 3-17 594. . h u d Code List P2, Phytoplankton, version 84334-GUZ, which N . A 1972: Observations o n phytoplankton Ill · k R &Retmer.C.W. t966: The dtatomsolt e nne 1 P .nne . . . e~~;ropiHed and non-~utr o phted archipelago waters ~f States. _ Vol. 1. 688 pp.. Philaddphta. was distributed as Annex IX of the Guidelines for the the southern coast "f Finland. - M ~morJnda so~ . Swift . E. 1913: Dissodintum pseudolunula n. 'P· - Fauna Flora Fenn1ca 4K·63- 74. Phy,ologta 12:90-91. .. Second Stage of the BMP (amendment to the Baltic Sea . A & Hallfor~. G . 1974: Some phytoplankton s pec l ~> W M 1952: Rockv-shore algae in the OreJ!.rund N1t:ml . · d s F na Flo ra fr,,m Balli~ water, . - Mcm\lran a oc. au a::~~hip~lago . _ Acta ·PhytogeoJ!.r. Suect~a 30: 1-29K. I- Environment Proceedings No . 12, 19 June 1985), should Fc nn~ea 49:77-93. XVI. F. ' 1 N' . !. SkuJa . H & Wdkn. T . 1970: Phyto pla nkto_n from 0 49· Entwurf etner nallirllchcn .tntct ung W atten b erg. H . I ., . . be substituted by the new list published in: lt:7;,1~ . P~JOV I k ~n - T'armlnne area. S. coast of Finland. dcr Ostscc. - Kiele r M~crcstors~h . 6: 10-15. . -Memoranda Soc Fauna Flora Fennlca 48.63-74 .. . ·k J l9'5· Ob~r c1nc Wa"crhlutc v11 n Hon T . & Chiha ra. M. 19gO: Re viSion ot the W o I oszyns a. . · · . N om s. R · E .. · B t Ma• Cyanoph y..: ~en on dcr Danztgcr Bud ll _und cone genus Tetrasc lmi' I Class Praslnoph y~eae) . - o . .,. Wucherung d~r D tatome~ Chactocerus Eobcnn . ~r un . -:- Gunilla Zetterberg (Tokyo) 93:317-339. . Bu ll. Acad. Polon . S..:. L ~tt r .. Cl. Math. Nat .. Scr. B. s,. H 1965: Bemerkungen zur dcr k System~ttk P an ow. · . . h b 1g oder Nat. (1): 101-114. pis. 6. 7. . . G . Code List P 4, Phytoplankton Anabaena-Formen mtt splraltg, sc rau • Zimmermann. B.. Moestrup. 0 . & Halltors. .. kreisformlg gewundenen T nchomen. - Llmnologlca Chrysophyte o r heliozoon: Ultra stru,tural stud tes ''n a version 86165-GUZ 1Berlin ) 3:1 63-172. cultured specoes of Pse udopcdinella (Pedtndla les ord. Pankow. H . 1976: Algennora der Ostsee. II. Plankton nov.). with comments on spcctcs taxonomy. - Published 1986, Stockholm (einsc hl. benth i~ h e r Kie~elalgen\ . - 493 PP·: Jena. . Protistologica (in press). . n p S 1''76: Check-list of Bnush manne Parke. M . & DI XO • . . ., ISSN 0282-8375

17~- PP, Received 14.V.1984

Contact address: 86165-GUZ, Kodcentralen, Riksmuseet S-104 05 ·Stockholm

6. Phytoplankton identification sheets

The phytoplankton identification sheets are under pre­ paration by the expert group convened by Dr. Ulrich Horstmann of the Federal Republic of Germany.

The phytoplankton sheets will be printed in the series Annales Botanici Fennici, Acta Botanic a Fennica, published by the Finnish Botanical Publishing Board, Helsinki. The reprint copies will be distributed to the Baltic Sea States through the Secretariat of the Helsinki Commission. -88- -89-

factors have been determined. Nano- and micro­ 7. Zooplankton zooplankton should be sampled by means of water For the Baltic Monitoring Programme (BMP), meso­ samplers or a plankton pump, macro- (2-20 em) and zooplankton ( 0. 2-20 mm) is the only obligatory megazooplankton (20-200 em) by means of horizontal or determinand. However, the undisputed significance of oblique hauls with larger nets (like the Bongo net), high speed samplers or small trawls (like the IKMWT, nano- (2-20 ~m) and microzooplankton (20-200 ~m) in the pelagic ecosystem justifies a recommendation to include Isaacs-Kidd Midwater Trawl) . them into the Monitoring Programme, but only as a tentative parameter (see also Section D.3.d) and D.3. b) Preservation Annex I). The samples shall be preserved in 4% formaldehyde solution (1 part 40% formaldehyde solution and 9 parts a) Sampling water). The formaldehyde has to be buffered to pH 8-8.2

Mesoplankton shall be sampled by means of vertical wi~h d~sodiumtetraborate (borax) (Na2B4o3 • 10 H2o). hauls with a plankton net (WP-2 net) . The WP-2 net c) Subsampling should have a mesh size of 100 ~m and should be hauled vertically with a speed of about 0. 5 m/s. The hauls shall be fractionated and the number of hauls to be For subsampling the whirling apparatus by Kott (2) or made at each station is dependent on local water the Folsom sample splitter (3) should be used. stratification. In principle, the following depth d) Species composition and abundance intervals shall be used:

When subsampling is needed, the volume of the subsample bottom - halocline (included) must be chosen so that at least 500 specimens are re­ halocline - thermocline (included) thermocline - surface tained. Copepods shall be analysed to developmental stage and sex of adult (c. I-III, c. IV-V, c. VI~ , The wire angle must be kept as small as possible during c. VI O). sampling to assure that the whole depth fractions are Since certain copepod nauplii and rotifers will pass sampled. the 100 ~m meshes, they can be excluded from the The use of sonars and pingers is recommended to get obligatory mesozooplankton analysis. exact knowledge of the depth position of the net. Macrozooplankton (medusae, mysids, chaetognaths, etc.) Flowmeters can be used, but they are non-obligatory. are not sampled adequately by the WP-2 net either and thus can be excluded from the obligatory analysis. Pumps and other sampling devices can be used if calibrated against the WP-2 net for the specific areas and for different times, and the proper conversion

7 483184A ~,r l -90- -91-

Macrozoobenthos e) Biomass 8. a) Soft bottom macrozoobenthos The biomass is calculated as the sum of individual The main responsibility for zoobenthos sampling and volumes. The list of individual volumes elaborated by analysis is as follows: BMB WG 14 (4) shall be used. Denmark: BMP K7, Pl, Ql, R3 Finland: BMP A2, A3, Cl, C4, Dl, F2, F5 f) Data reporting German Dem. Rep.: BMP K4, K7, K8, M2 Fed.Rep.Germany: BMP Nl, N2, N3 Poland: BMP Kl, K2, Ll Data shall be reported on standardized formats Sweden: BMP Il, K2, K4, R6, R7 Soviet Union: BMP F2, F5, J2, Kl (according to Sections D.lO and D .11) . The abundance and the biomass should be expressed both per m' and per b) Soft bottom macrozoobenthos analysis

m3 • If flowmeters have been used, both the original and the corrected values shall be listed. For the purpose of the Baltic Monitoring Programme, the macrofauna is defined as that part of the soft bottom g) References in Chapter D.7. fauna which is retained on a sieve with a mesh size of 1.0 x 1.0 rom (see 8). 1. sieburth, J. MeN., Smetacek, V. & Lenz, J. 1978. Pelagic ecosystem structure: heterotrophic compartments of the plankton and their Sampling relationship to plankton size fractions • Limnol. Oceanogr., 23: 1256-1263. Sampling on shallow stations (70 m or less) should pre- 2. Kott, P., 1953. Modified whirling apparatus for the subsampling of plankton. Aust. J. Mar· ferably be conducted during the daytime, since some Freshw. Res., 4 (8): 387-393. benthic species have semipelagic activity during the night. It is essential that sampling of macrozoobenthos 3. McEven, E.F., Johnson, M.W. & Folsom, T.R. 1954. A statistical analysis of the performance of the is accompanied by some hydrographic sampling to provide Folsom plankton sample splitter based upon test observations. Arch. Met. Geophys. information about the hydrographic situation. There­ Bioklimatol. Ser. A, 7: 502-527. fore, as a minimum requirement, water should be sampled as close as possible to the sea bottom, or a profile 4. Hernroth, L., (ed.), 1985. Recommen~atio~s on methods for marine biological stud1es 1n the with a calibrated CTD and oxygen probe should be taken. Baltic Sea. - Mesozooplankton biomas~ assess­ ment by the individual volume techn1que. BMB Publ. No. 10. 32 PP· The widely applied 0.1 m' Van Veen grab (modified version after 2) has to be used as the standard gear for benthic macrofauna sampling in the Baltic Sea, because of its comparative reliability and simplicity of handling at, sea. The grab should weigh about 25-35 kg when emptied. In order to reduce the shock wave caused by lowering tne grab, the windows on the upper side shall cover an area as large as possible, in practice around 60% of its upper surface. The windows -93- -92-

The choice of sample size and number of samples is shall be covered with metal gauze of 0.5 x 0.5 mm mesh always a compromise between the need for statistical size. accuracy and the effort which can be put into the study. One way to do this is to calculate an index of There may be cases where the use of other gear with precision. The ratio of standard error to arithmetic smaller sampling area is advisable, e.g. if the fauna mean may be used ( 3) , i.e. ( x = arithmetic mean, s = is very dense and uniform. When other gears than the standard deviation, n = number of samples). A standard grab are employed, intercalibrations have to reasonable error would probably be ::_ 0. 2 -"- 2 0%. be done on a regional basis and on specific sediments on which these samplers will be used. When a change of gear is intended, it is recommended to sample parallel / D = (D L 0.2) with both gears for a period of 3-5 years.

Means are to be provided for attaching 20 kg of lead to On the representative statl' ons, at 1 eas t f l· ve samples the upper edges of the jaws or inside the grab. should be taken to enable the investigator to reach a certain level of precision by sorting as many samples Precautions that must be taken when using the grab: as necessary. The same procedure is strongly rec- ommended for all other benthos stations unless another The settling down and the closing of the grab must sampling strategy (area sampling) is employed in be done as gently as possible. This will reduce the national/coastal monitoring programmes (see also shock wave and the risk of sediment loss as a result Sections A.l and A.2). of lifting the grab before completed closure, The depth to bottom must be recor d e d and documented as possible to The wire angle must be kept as small separately for every sample at the time of sampling. and lifted up ensure that the grab is set down Nevertheless, the depth range shall be kept as small as vertically. possible.

If, as often happens on sandy bottom or erosion Each laboratory shall carefully check the exact sediments, less than 5 1 of sediment is collected, the sampling area of its grab l. n order t o rna k e possible a sample should be regarded as not quantitative, and a correct calculation of the number of individuals per new sample should be taken after loading the grab with square metre. an extra 20 kg of lead as described above. This may as much as double the effective sampling depth of the Sediment description shall be given by visual grab. If less than 5 1 of sediment is still collected, observation stating sediment type, colour, and the the sample may be used, but the low sample volume approx. depth .of the oxygenated surface layer (Section should be stressed when results are given (cf. 2, p. D.lO, last page). 64). The evidence of this problem may be different in different parts of the Baltic Sea, depending on, e.g., how deep in the sediment the species live. -94- -95-

All residues retained on the sieves should be Sieving carefully flushed off the sieves with water from The standard sieve for the Baltic Monitoring Programme below and fixed. Spoons and other tools for sample shall be of metal gauze (stainless steel, brass or transfer should be avoided; bronze) and have a mesh size of 1.0 x 1.0 mm. In order When the 0.5 x 0.5 mm sieve is used, the 0.5 and 1.0 to collect quantitatively developmental stages of the mm sieve fractions must be kept separate throughout macrofauna and abundant smaller species it is, however, all further processing. recommended to use an additional sieve with mesh size of 0.5 x 0.5 mm. This sieve must have the same Other sampling methods qualities as the 1 mm sieve. The mesh size of th~~­ sieves has to be checked from time to time for damage Dredge hauls may be valuable as a complement to grab samples, since mobile as well as large but compara­ and wear. tively rare species are more easily caught by dredging. Attention must be paid to the following points: This is · especially true in areas almost devoid of benthic fauna, where grab samples may be too small to - Each sample must be sieved, stored and documented collect the remaining specimens. Dredge samples may separately; also be used for describing the relative dominance. - The volume of each sample must be measured. This can Descriptions of suitable dredges can be found in (4). be done by grading the container or by using a ruler; - The grab has to be emptied into a container and should be brought portion by portion onto the sieve In areas where the burrowing depth of the fauna are as a sediment water suspension. The use of beyond the penetration depth of the grabs (or that type sprinklers and hand-operated douches to suspend the of gear cannot be used), core samplers may be advisable sample is recommended. Very stiff clay can be gently to use, provided that their efficiency has been satis­ fragmented by hand. Between the pourings the sieve factorily proven by intercalibrations. (see Section must be cleaned to avoid clogging and thus to ensure D.8.b)). an equal mesh size during the whole sieving procedure; In order to survey a large area and interconnect point­ - The sieving of the sample has to be done carefully in like (station) information on epibenthos, large-scale order to avoid damage of fragile animals. Therefore, survey methods such as side-scan sonar and under­ a direct jet of water against the sieve should be water-TV may be used. avoided; - Visible fragile animals, e.g. some polychaetes, shall Fixation be hand-picked during the sieving; stones and big shells should be picked out to avoid the grinding The hand-picked animals and the sieving residue shall be fixed in buffered 4% formaldehyde solution (1 part effect; 40% formaldehyde solution and 9 parts water). It should be noted that formaldehyde is regarded as toxic and -96- -97- probably carcinogenic. Therefore, it should be handled Analyses with great care and means for waste air exhaustion should be provided for all laboratory procedures. For Species composition, abundance and biomass should be buffering, 100 g of hexamethylenetetramine (Hexamine = determined. Urotropin) shall be used per 1 dm 3 of 40% formaldehyde. Sorting Sodiumtetraborate (= Borax) in excess may also be used. Sorting should always be done using magnification aid Preservation and storage (magnification lamp, Stereo-Microscope). Small portions of the unsorted material shall be put on a 0.5 mm mesh The samples shall be stored in dark jars or in the size sieve and washed with tap water. The portions must dark. The pH of the samples is to be checked every be flushed into a transparent glass dish of about 20 em third month and adjusted to about pH 7 by the addition diameter covering the bottom of the dish as only a of buffer if necessary. lo?se and semi-transparent layer. The material shall be looked through twice by regular scanning. The dish must Staining be shaken before the repeated search.

In special cases, i.e. samples from sandy bottoms, it During the first examination, a piece of dark foil may be advisable to stain the 1 mm sieve samples to shall be placed under the dish while a piece of light­ facilitate the sorting process. coloured foil shall be used for the second examination. This ensures equal visibility of light- and dark­ When the 0.5 x 0.5 mm sieve is used, this size fraction coloured animals. shall always be stained with Rose Bengal for more effective sorting. The staining shall be done before After having sorted each sample (except the first!) the sorting by: index of precision (3, p. 129; see also the above paragraph on sampling) has to be calculated. When - washing the sample free from the preservation fluid reaching the proposed level of precision ( D::::, 0. 2 ~ 2 0%) by using a sieve with a mesh size smaller than 0.5 x no further sample from this station needs to be sorted. 0.5 mm; This applies only for the dominant species, each of allowing the sieve to stand in Rose Bengal stain which takes up more than 10% of the total abundance. (1 g/dm3 of tap water + 5 g of phenol for adjustments to pH 4-5) for 20 minutes with the sample well When the 0.5 x 0.5 mm sieve is used, this size fraction covered. shall be sorted by scanning the sample twice under a stereomicroscqpe. Before sorting, the size fraction However, Rose Bengal (4 g/dm3 of 40% formaldehyde) may could be subsampled using a subsampler described by be added already to the fixation fluid. Dybern et al. (2, page 61). -99- -98-

determined after incineration at 500-520°C in an oven Broken animals shall only be counted as individuals by until weight constancy (about 12 hours, depending on their heads (e.g. polychaetes) or hinges of bivalves sample and object size). The temperature in the oven with adhering pieces of tissue. should be checked with a calibrated thermometer, because there may be considerable temperature gradients Biomass determination (up to 50°C) in a muffel furnace. Caution is advised not to pass a certain temperature ( 550 o) since then a The biomass shall be determined as formalin wet weight, sudden loss of weight may happen due to the formation formalin dry weight and ash-free dry weight. of CaO out of the skeletal material of many invert­ ebrates ( caco ). This can reduce the weight of the The biomass determination shall be carried out for each 3 mineral fraction by 44%, This decomposition occurs very taxon separately. abruptly and within a small temperature interval ( 6 ) • During the initial period after fixation the biomass of organisms changes, and therefore formalin wet weight Before weighing, the samples must be kept in a desic­ shall be measured not earlier than three months after cator, while cooling down to room temperature after preservation (1). drying as well as burning. Ash-free dry weight may also be calculated using the conversion factors published by The formalin wet weight is obtained by weighing after BMB ( 5) • external preservation fluid is removed on filter paper. While removing the water, the larger animals shall be c) Data reporting carefully and delicately turned over on the filter paper. The animals are left on the filter paper until All data should be reported according to details given no more distinct, wet traces can be seen. In practice, in Sections D.lO. and D.ll. tiny animals dry within a few seconds, while larger ones take 20 seconds or longer to dry. Shelled animals Whenever some taxon found on the sieves is excluded are weighed together with their shells. In the case of from the reported results, this should be explicitly large bivalves (length over 15 mm), the two shells are stated and the reason explained (e.g. Piscicola opened with a scalpel and laid on filter paper. The geometra not included because it is a parasite) on the opened shells must be squeezed so that all the water Plain Language Record. Also results from dredge hauls flows out of the mantle cavity. As soon as the non­ and any other survey methods should be reported in the tissue water has been removed, the organisms are Plain Language Record. weighed with an accuracy of 0.1 mg.

A sediment description can be given in the Plain The formalin dry weight shall be estimated after drying Language Recor'd or, ~~f a more comprehensive sediment the formalin material at 60-80°C to constant weight analysis has been made, in the reporting format for (for 12-24 hours, or an even longer time, as may be contaminants in sediments (see Section c.III). necessary for this process). Ash-free dry weight should be estimated after measuring dry weight. It is -100- -101- d) References in Chapter D.8. 9. Micro-organisms 1. Brey, T. 1986. Estimation of annual P /B-ratio and production of marine benthic invertebrates from Methods to be used for microbiological monitoring length-frequency data. OPHELIA, Suppl. 4: 45-54. Total number and biomass of bacteria H., Elmgren, R. 1976. 2. Dybern, B.I., Ackefors, Production of bacteria Recommendations on methods for marine biological studies in the Baltic Sea. BMB. Colony-forming bacteria (colony count) Publ. No. 1, pp. 63-76.

3. Elliott, J.M. 1977. Some Methods for the Sampling sites and frequency Statistical Analysis of samples of Benthic Invertebrates. Freshwater Biological Association Scientific Publication No. 25, On monitoring cruises, including standard stations, 158 pp. microbiological measurements should be performed on the 14 4. Holme, N.A. & Mcintyre, A. 1984. Methods for the same samples as c-primary productivity measurements Study of Marine Benthos. Oxford, 387 pp. are made~

5. Rumohr, H., Brey, T. & Ankar, S. 1987. A for compilation of biometric conversion factors For baseline studies near-shore stations (at least one, benthic invertebrates of the Baltic Sea: BMB Publ. No. 9: 69 pp. but preferably three) should be monitored with higher frequency, at least once a month. 6. Winberg, G.G. 1971. Methods for the Estimation of Production of Aquatic Animals. Academic Press. London, 175 pp. Sampling 7. Zetterberg, G. 1988. Code list 01 Baltic invertebrates, version 88163-GUZ. Kodcentralen, Water for microbiological analyses should be taken from Riksmuseet, Stockholm, S-10405. the same samples as for primary production and revised 2nd 8. BMB Publication No. 12, in prep. = ch lorophy 11-!:;. edition of the BMB.

The standard sampling depths for microbiological measurements are: 1 m (2 m), ( 3 m), 5 m, 10 m, (15 m) and 20 m, non-obligatory depths being given in brackets.

Sampling should preferably be performed in the morning.

Data reporting

All data repo'rting should follow the biological data reporting format and should be given along with the phytoplankton determinands' (primary productivity, chlorophyll-!:; and phytoplankton): -103- -102-

Procedure for bacteria counts: total number, mean cell volume and the factor for converting cell volume to carbon Shake the sample well and draw a 20 ml subsample from should be given, the sample bottle. for production of bacteria: thymidine incorporation and factors for calculating net production should be Add 0. 5 ml of formalin into the 20 ml sample through given, the sterile filtration unit. Let a few drops out of the for colony-forming bacteria: the number of colony unit before starting to add it into the sample vials. forming bacteria per ml of water sample should be

given. Store the samples in the refrigerator for further processing. Good preparations are obtained if staining a) Method for measuring total number and biomass of is done within 24 hours after formalin fixation. bacteria However, good preparations have also been obtained after storage times of several months. The method for measuring total number and biomass of bacteria with Acridine Orange (AO) staining follows AO-solution essentially the method presented by Hobbie et al. ( 2) • However, one should note that other stains, DAPI (3), 1. Acridine orange (AO) solution: 0.5 mmol ACRIFLAVIN (1) etc. may work better than AO in different types of waters. 15.1 mg of AO + 98 ml of deionized or distilled water + 2 ml of unbuffered formalin (35-39%) Acridine orange direct count (AODC)

- make the solution particle-free by filtration with Materials a disposable filtration unit (pore size 0.2 ~m) just prior to staining as described below, or Scintillation or other glass vials (particle free, heated in 500 °C for 4 hours) for collecting and make the solution particle-free by filtration, storing the samples cover it with aluminium foil (solution is light Formalin (35-39%), unbuffered sensitive) and store it in the refrigerator (may be Sterile filtration units for making particle-free stored up to several weeks). liquids Syringes 2. Irgalan black solution Membrane filters Nuclepore filters (pore size 0.2 ~m) 2 g of Irgalan black/one liter of deionized water. Add 20 ml of acetic acid to make the solution 2% acid. ..,.-"'~-'*~-- ' -105- -104-

Take off the Nuclepore filter when vacuum is still on Irgalan black staining of Nuclepore filters and write with a pen at the edge of it: sample label and volume of filtration. Stain the Nuclepore filters (25 rom, pore size 0.2 ~m) 30 min. in 0.2 ~m filtered Irgalan black. Rinse care­ Allow the filter to dry for a few minutes (5-10 min) fully 3 times on 0.2 ~m filtered deionized water. Dry and place it back into the box. the filters (5-10 minutes) and place them back in their particle-free petri dishes for case. Use clean, Store the filters in a dry, dark place (may be stored staining and rinsing. for several months).

Sample preparation Microscopy

Rinse the filtration system thoroughly with sterile Place a drop of immersion o'l~ ( n d .. 1 515) on the water to start the procedure. objective glass.

Place the Sartorius, Millipore etc. membrane filter Place the filter (or a part of it if you want to look (pore size 0.2 ~m) on the sinther as the supporter. at it again later) on the oil so that air bubbles are not caught under it. Place the Irgalan black stained filter on the wet

Sartorius: avoid air bubbles. Place a drop of immersion oil on the filter.

Staining procedure Place a cover slip on the filter without creating air bubbles. Add 5 ml of particle-free deionized or distilled water

into the funnel. Place a drop of immersion o'l~ on t h e cover slip. The sample is ready for counting. Pipette l-2 ml of sample water into the funnel (mark the date and filtered volume on the sheet) and filter Count at least 20 fields (10-20 cells/field). For to dry. Add 2-5 ml of particle-free deionized or calculations, you need the filtration surface area and distilled water into the funnel and continue filtering the area of the counting (equation 1). to dry. ( 1) N (cell/ml) = X * A * d * 106 * a-1 * n-1 * v-1 Stain the sample with 300-500 ~l of particle-free AO solution (0.5 mmol). Use the sterile filtration unit. A = area filtered (e.g. 193.59 mm2) Keep the filtration unit covered with aluminium foil. d = dilution due to formalin (20.5/20) = 1. 025 a = area counted (e.g. 640 ~m' ) Allow two to five minutes for staining with AO. n = fields c'ounted (e.g. 20) v = volume filtered (1-2 ml) Allow the filter to dry. X = total count of cells -107- -106-

References: Cell volume determination 1. Bergstrom,c . I., Heinanen ' A · & s a 1 onen, K. 1986. Bacterial cell size distribution is determined by cali­ <;>mpar~s<;>n. of Ac;ridine Orange, Acriflavin and ~~sbentz~m~n sta~ns for enumeration of bacteria brated ocular graticules e.g. Patterson-Cawood or New ~~ cl:ar and humic waters. Appl. Environ. Forton Grids (Graticules, Ltd., England). At least 20 M~crob~ol. 51: 664-667. fields and the total of at least 50 cells are measured 2. Hobbie, J.E., Daley, R.J. & Jasper S 1977. Use of (classified). The cells measured must be selected Nuclepore filters for counting. bacteria fluorescence microscopy A 1 by randomly, e.g. count the cell closest to a pre­ Microbiol. 33: 1225_1228 • · PP • Environ. determined spot of the graticule from each field. Do 3 • Port:r, K:G •. & Feig, Y.S. 1980. The use of DAPI for not hesitate to count 50 fields altogether. ~dent~fy~ng and counting aquatic microflora. L~mnol. Oceanogr. 25: 943-948. The actual measurement is done with the aid of different size circles on the graticule. For cocci . b) Production of bacteria select the circle closest to the size of the organism: With Patterson-Cawood grid, and with about 1200 times Method for measuring bacterial net production using magnification the smallest circle is close to 0.2 ~m. Tritiated Thymidine Incorporation For rods pick the circle that is closest to the width (W) of the rod and another circle that is closest to Bacterial production is measured by a modified the length (L) of the rod. TTI-method (Tritiated Thymidine Incorporation), Fuhrman

and Azam ( 1) ( see F ~gure· D • 2 •) • This procedure was Cell volumes are counted according to equation 2. adopted for routine use by Larsen --et al. (2).

Biovolume ~m3 = ( n/4)W2 (L-W/3), where ( 2 ) Materials

L = cell length 3 H-methyl-thymidine (20-100 Ci/mmol) W = cell width Formalin (35-39%) For cocci L = W. Trichloroacetic acid 5% (ice-cold) Filtration unit (e.g. Sartorius SM 16 547 or Millipore Results 1225 Sampling manifold) Membrane filter (0.2 ~m) Results must be expressed as: Liquid scintillation equipment

6 A. Total number of bacteria, cells ml-l *10 (l.OOE6) " -1 B. Mean cell volume, ~m 3 cell , and coefficient of variation (l.OOOE-3), CV% c. Factor for converting cell volumes to carbon = -3 0.35 pgC ~m

Biomass of bacteria = A * B * C -109- -108-

Procedure FIGURE 0.2. Flow chart of TTI-measurement technique, sample volumes, incubation times and liquid scintillation procedure may vary Twenty to twenty-five ml water samples are incubated 3 with 10 nM tritiated thymidine ( H-methyl-thymidine, between sites. 20-100 Ci mmol-1 ) for 30 minutes at in situ tempera­ ture. When temperature is below 5°C incubation up to 2 hours may be used. 25 ml seawater 10 nM 3H·thymrdine Incubations should be started immediately after sampling. If storage cannot be avoided, larger amounts t of water (e.g. 1 1 sample bottles) must be kept at in 30 min incubation situ temperature, in a water bath, in the darkness and incubations should be started within an hour.

t Three formalin-killed (1% final cone.) blanks are 0.8 ml 37% formalin incubated along with three sub samples. Allow 10 min. 500J.ll ethyl· before adding label to blanks. Incubations are stopped acetate 30 min 10 ml by adding formalin to a final concentration of 1%. 10 ml Aqualyte icecold ""'- J Formalin-killed samples are stored in the refrigerator 5% TCA ~ until filtration. A known volume of the samples is ··················0.2}-Jm filtered through 0.2 ~m membrane 25 mm diameter filters (Sartorius SM 111, 113, Millipore, Asypore). The filters are rinsed five times with 1 ml ice-cold TCA (trichloroacetic acid) and again five times with 1 ml TCA with the chimneys of the filtration device removed.

The filters are stored in glass (preferably) or plastic vials and, prior to liquid scintillation counting, solubilized by 30 minutes' treatment with 0.5 ml ethyl acetate. Certain scintillants (e.g. PCS, New England) may make use of a solubilizer unnecessary (with Millipore BS, Sartorius 113 and Asypore). This alterna­ tive is to be preferred since ethyl acetate is a strong quenching agent and therefore inaccuracies in pipetting will introduce large errors in counting efficiency. ''-<;;:;r:yiJ/i(% ,, .,.Ji~w,. ,._j;l. ~'·

-llO- -lll-

After addition of the scintillation cocktail, the Net production is thus calculated according to: 3H-activity is assayed by liquid scintillation counting and corrected for quench by external standard channels Net production, mgC m-3h-l = TI * CF * cv * cc ratios. Formalin pre-fixed samples are used as blanks. 1 Thymidine incorporation (TI, mol 1-l h- ) is calculated -1 where TI = thymidine incorporation, mol 1 h-1 as: CF = 1.1 * 1018 cell mol -1 3 cv = !lffi cell-l 7 -3 dpms - dpmB cc = 3.5 10- !lgC !lffi TI = * SV X T X SA

3 where dpms and dpmB are the H-activities of the sample References and blank, respectively, SV is sample volume {1), T is 1. Fuhrman, J.A. and Azam, F. 1982. Thymidine incubation time (h), and SA is specific activity of the incorporation as a measure of heterotrophic 3 1 bacterioplankton production in marine surface H-thymidine (dpm mol- ). waters. Mar. Biol., 66: 109-120.

2. Larsen, J.B., Bjornsen, P.K., Gaertz-Hansen, o. and Batch cultures to evaluate the conversion of thymidine Olesen, M: 1985. Provisional investigations on into bacterial cell production should be performed for the pelag1c carbon flow in open Danish waters. Rep. Mar. Pollut. Lab., 8: 1-49. each new sampling site during each of the annual suc­ cession stages (e.g. spring, summer and winter). For a 3. Riemann, B., Bjornsen, P.K., Newell, s. and Fallon, R. 1987. Calculation of cell production of discussion on this subject, see (3). coastal bacteria based on measured incorporation of H-thymidine. Limnol. Oceanogr 32: 471-476. • For batch cultures, seawater (250 ml) is filtered aseptically through a 0. 22 11m Millipore filter. As an inocculum 25 ml seawater, filtered through 0. 6 11m to c) Colony-forming bacteria remove predators, is added. The batches are incubated at in situ temperature for 48 h. Every 12 h subsamples Determination of the number of colony-forming bacteria are taken and preserved with formalin for bacterial enumeration by AODC. TTl-measurements are carried out Introduction at 4-8 h intervals following the procedure described above. Finally, for each 12 h period the integrated The determination of the number of colony-forming thymidine incorporation is compared with the observed bacteria is routinely performed by most of the increase in cell numbers to obtain an empirical con­ microbiological laboratories. These measurements do not' version factor from thymidine incorporation to cell require special apparatus other than basic equipment, production. As a guidance, a conversion factor of 1.1 x The number of colony-forming bacteria is an easily 18 10 cells mol-l thymidine incorporated may be applied obtainable parameter which gives some information on the degree to coastal marine environments ( 3 ) • The measured of pollution of the water bodies undo 1 3 study. It average cell volume and a factor of 0. 35 pgC 11m- has been found that the colony-forming should be used to convert production data into carbon terms. -112- -113-

Thermostatically controlled water bath bacteria often react rapidly to environmental stress the - The flasks containing the nutrient medium should be ( 2, 3 ) • Thus, in eutrophied and polluted areas, also its maintained at 42°C. number of colony-forming bacteria and percentage of the total bacterial number are subs- tantially higher than in clean waters. Nutrient medium ZoBell's medium 2216 E is suggested. This medium since the number of colony-forming bacteria is affected contains 5 g of peptone, 1 g of yeast extract and 15 by environmental changes, this implies that also the g of agar per liter. The pH of the medium should be type of nutrient media, the incubation temperature, 7.6. etc. influence to a great extent the results. The - Since the composition of peptone, yeast extract and rigid agar may vary according to the supplier, the products method, therefore, necessarily requires a from DIFCO (Detroit) should be used. standardization. The water used for preparing the medium should be

For the determination of the colony-forming bacteria ~ged sea water with a salinity close to that of the the ''pour plate" and the "spread plate'' methods exist. water samples to be studied. In most cases the results obtained by the two methods - It is convenient to put 10 ml portions in test tubes, are quite similar (4). Since the ''pour plate" method is since the test tubes allow rapid liquefying of the more frequently employed in routine measurements, it is medium and afterwards fast maintenance at 42°C. The suggested that it be used for microbiological medium should be sterilized for 20 minutes at 12l°C in an autoclave. monitoring (1).

Incubator Equipment - The incubator should maintain a temperature of 20°C.

Microbiological water sampler - ZoBell-samplers are very convenient. Colony counting device

Sampling depths Dilution vials - Screw cap test tubes with 9 ml of sterile water having a salinity close to that of the sample may be The water samples should be taken with the microbial water sampler under sterile conditions from 3 m below used. the water surface, 3 m above the sediment, and at the depth of the halocline. The depths should coincide with Sterilized 1.0 ml pipettes those for phytoplankton and chemical analyses.

Petri plates of 9-10 em ¢ Storage

Storage of water samples should be avoided. A storaqo time of 3 hours, however, in a refrigerator may bo tolerable. -114- -115-

5) When the agar medium has cooled down to room Procedure temperature, the dishes are placed upside down and The whole procedure has to be done under aseptic condi­ put together into stacks. These are covered with plastic bags and then incubated at 20°C in the tions. incubator. 1) samples from the chosen depths are taken and 6) The colonies are counted after 7 and 14 days of thoroughly shaken. incubation. The first counting is considered only in 2) The number of colonies growing on the plates should the case of fungal growth, which usually does not be between 30 and 300. It may, therefore, be appear before the second week of incubation. necessary to dilute the water samples with sterile water, with the salinity close to that of the sample 7) The results are reported as number of colony-forming bacteria per ml of water sample. water.

The appropriate dilutions are prepared by pipetting References 1 ml of the sample to the first dilution vial and 1 ml from there to the second vial and, if 1. Gunkel, W. and G. Rheinheimer (1968): Bakterien, PP • 142-157. In: Methoden der meeres­ necessary, continuing to the third vial. Mix well bGiologischen Forschung (C. Schlieper 1 Ed.) between the dilution steps. Thus, dilutions up to ustav Fischer Verlag, Jena. 10-3 are obtained. 2. Rheinheimer, G. (1977): Regional and seasonal distribution of saprophytic and coliform bacteria •. In: Microbial ecology of a brackish In near-shore regions, dilutions of the samples of water envlronment. Ecol. Studies, 25: 121-137. 10-2 and 10-3 may be necessary, whereas in the open Ed: by G. Rheinheimer, Springer Verlag, Berlin, Heldelberg, New York. -1 dilution Baltic Sea the original sample or the 10 3. Rheinheimer, G. (1984): The role of small· may be used. h eterotrophs (bacteria and protozoa) in a shelf ecosystem. In: The Biological Productivity of 3) 1.0 ml portions of the water sample or of a suitable North :Atlantic Shelf Areas. Rapp. P.-v. rieun. Cons. lnt. Explor. Mar. 183: 144-151. dilution is pipetted into 3 sterile Petri dishes. 4. Vii.\iti'inen, P. (1977): Effects of composition ,,r· substrate and inoculation technique on pltll.

Section 3.3 Chlorophyll a 10. Biological data reporting format 3. 3. 1 TzEe Master Record (pp. 9 and 25 1n July 1981 version and In this-version)~-

Column 53 bas been assigned to "Calculation method". International Council for the Exploration of the Sea Section 3.4 ~oulankton Revised August 1983 3. 4.2

REVISIONS TO THE BIOLOGICAL DATA REPORTING FORMAT Column 24: a neY code number has been added for the RUBIN Code List PL, Phytoplankton, limnic.

In relation to the July 1981 version of the Biological Data Reporting Format, Section 3.5 Mesozooplankton the folloYing changes have been made: Q~!~_Qz£!~-~~£~~~ (pp. 14 and 30 1n July 1981 version Section 3.2 Phytoplankton Primary Production and in this version). (pp. 6 and 23 1n July 1981 version 3. 2. 1 TiE~-~~~!~~-~~£~~~ Column 24: a neY code number has been added for the and in this version). RUBIN Code List 01 Baltic invertebrates. Column 53 bas been a.ssigned to "Attenuation determination Column 37 should be changed to unassigned from identifier. method". Column 38 should be changed to "D eve 1 opmental stage and Columns 54 to 72 are unassigned. sex". The assignment of columns 70-73 to "Total irradiance (in situ)" and columns 74-77 to "Total irradiance (expe­ Section 3.6 Zoobenthos rimental)" bas been deleted, and 3.6.2 Column 73 bas been assigned to "Irradiance unit". 2~!~_Qz£!~-~~£~~~ (p. 18 in both versions).

Columns 74-77 have been assigned to "Incubator irradiance". Column 24: a ne•• co d e num b er bas been added for the RUBIN Code List 01 Baltic invertebrates.

3. 2. 2 Data Cycle RecordWP· 7 and 24 1n July 1981 version and In-tti5-¥e;si0n1~ Columns 24 to 27 have been assigned to "Relative ura­ diance". Columns 45-49 are noY assigned to "Potential prl!:Jary pro­ duction" (addition underlined).

Columns 50-54 are noY assigned to "Daily production ~ m3" (addition underlined). Columns 60-62 are now assiqned to "Carbon d1oxide".

Columns 70-74 have been assigned to "Daily production per m2". -119- -118-

INTERNATIONAL COUNCIL FOR THE EXPLORATION OF THE SEA CONTE1'TS BIOLOGICAL DATA REPORTING FORMAT

1. mRODUCTION •.... · . · · · · • · · · • · • · · · · · • • · • · · · · · • · · · • · · · · · · • · · · · · · · · 1. INTRODUCTION 2. OVERVIEvl OF THE SYSTEM • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • 6 This format has been developed in response to a request to the International ). DErAILED DESCRJ:Pl'ION · · · · • • • • • • • · • • · • · • • · • • • • • • · • • • • • • • • • • · • • • • • • Council for the Exploration of the Sea by the Interim Baltic Marine 6 Environment Protection Commission for a data reporting format which could Bio:t'Bster Record . ·. · · · · · · · • • · · · · · · · • · · · · · · · · · · • • • • · · · · • · · · ).1. 9 be used for biological measurements obtained in the Baltic Moni taring Phytopl~~ton Primary Production ··•••••·•·•···••••••••·••• Programme. The format has been designed to interface with the ICES ).2. 12 Chlorophyll .§:. •••••••••••••• • • • •• • ••• • ••••• • •••••• • •••••• •. Oceanographic Data Reporting System (see Manual on ICES Oceanographic ).). 14 Punch Cards, 4th Ed. 1979), which is used to report hydrographic and hydrochemical data relevant to an interpretation of the biological results, ).4. Phytoplankton • • · • • · · • • • • • • • · • · • • • • • • • • • • • • • • • • • • • · • • · • • • • • 16 •••••••••••••••••••••••••••••••••• 0 ••••• and has been based on the IOC GF-3 format for the international exchange of Mesozooplankton ).5. 20 oceanographic data.

•••••••••• 0 ••••••••••••••••••••••••••• 0 0 •• 0 0 •••• 3. 6. Zoobenthos 23 3. 7. Plain Lang'\lage Record ...... • • • · • · · · · • · · · · · · · · · · · · · · · The Biological Data Reporting Format is intended to be flexible and is 24 amenable for recording and exchange of data via standard forms, punch GENERAL REMARKS FOR ALL RECORDS ••.•••••••••••••••••••••••••••••• cards or magnetic tapes. 4· 2. OVERVIE\1 OF THE SYST&'l APPENDIX 1 -!DC COUNTRY CODES FOR ICES MEMBER COUNTRIES .. ·•·••••• 25 Four types of records are included in this format: a File Header Record (the BioMaster), a Series Header Record (the Type Master), a Data Cycle Record, and a Plain Lang~age Record (see Figure RECORD FORMS : 1). 26 J3i oMas ter Record ...... • . · · · · · · • · · • · · · • • · · · · · · · • · • • · · · · · · Phytoplankton Primary Production 27 FIIE HEADER RECORD Type Master Record •• 0 •••••• 0 • 0 •• 0 •••••••• 0 ••• 0 •• 0 • 0 0 •• 0 ••••• 28 •• 0 0 ••••••••••••••••••••••••• 0 ••••••••••••• BIOMASTER Data Cycle Record Chlorophyll §!; 29 Type Master Record ...... •. • • · · · · • · · • · • · · • · · • · · · · • · · · · · · · • • 30 Data Cycle Record .... • .. • · • · · · · • • · · · · · • · • · · · · · • · · · · • · • · · • · • • Phytoplankton 31 Ty-pe Master Record ... • ... · • · · • • · • • · • • · • · • • · · • · • · • • · · • · • · · • • • SERIES READER SERIES READER SERIES READER 32 Data Cycle Record ..... • .. • • • • · • • · • • • • • · • · • · · • • • · · • • • • • • • · · · • RECORD RECORD RECORD Mesozooplankton 33 TYPE MASTER TYPE MASTER Type Ma.ster Record ...... •... • • • · · • • • • · • • · • • · • • • • · · • · • · • • • • · 34 Data Cycle Record ...... • .. • • • · • • • · • · • • • · · · • · • · · · • • • · • · · • · • Zoo benthos 35 Type Master Record ...... •. • · • • • · · • • · • • · · • • · · · · • · • · · • · · · · · • · • 36 Data Cycle Record ...... • · · • • · • · · • • • · · • • • · · · • • • · · · · • • · • · • · • • • 37 Plain Language Record ...... • . • · · · · • · · • · • • · · • · • · · · · • • • · · • · · • • • • 38 I \ i \ Sample Description Form ...... · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · DATA CYCLE DCR DCR DATA CYCLE DCR DCR DATA CYCLE DCR D8R DCH

- 0 - 0 - RECORD RECORD RECORD

FIGURE 1: OVERVIEW OF FORMAT -120- -121-

Note:

For the reporting of data on mesozooplankton, the form labelled "zooplankton" should be used. When data on protozooplankton

~ ~ >-'· ~ (tentative parameter) are to be reported, the "phytoplankton" Country ...' f "' Country '' I Country v<"' 00 v<"' reporting form should be used and the Phytoplankton Type ~ " Ship ' Shi _,. Ship "' I "" p ~ Master Record should be marked with the code Z in column 77. ~"' f..n 0''"" '"" a-, 0 __, Station No. ' ~Station No. .L __, Station No. 0 } " (J) CD CJ) " ~ -D -.[) l 0 ~ rl- >-' ,__, <+ "'.... 0 Year b Year .... 0 -- ~ ------rl- >-' } >-' ~ ~ >-' >-' ~ "0 ,.., >-' >-' ~ "" .... Parameter Parameter p. "' "' "' ~ ::;; i ::;; ....<+" ',;:' Method ( ',;:' Jo'iethod ',;:' 0 t-< ...,.... ' 0 Specification ' Specification 0 ~ ( ~ I•'•" -~ ----- "'d <+ ~ r:: ...."' 0' p, 0 ~ "' 0" 0 i:j ~ ~ CJ) NSEW "'<+ ~ § tl> !:;:; "0 "' '< ~ 0 Ol 0 Year "' ~ <+ "' <+ .: ~ (D "' (D (D '"' "'0 0 "'0 " >; 0 tl> '"'0 0 0"" " >; >; p. ""

,_, >-' .... tl> "" "-' < .... Q (J) "' I~ ~ [J)" Ol " ,., g ,,no

:~J' ((> ~ n "'(D 0,, p, (D "'0 0 >; p.

9 483184A -122- -123-

The BioMaster Record serves as the file header for the full series of BioMaster Record biological data obtained at one station. The first 27 columns of the Column BioMaster Record are identical to the first 27 columns of the cards used Item Code Description in the ICES Oceanographic Data System so that the relevant hydrographic, hydrochemical and meteorological data may be easily correlated with the 1-2 Country IOC Country Code (see Appendix 1). biological data obtained at the same station. The remainder of the Ship BioMaster Record is used to record the types of observations made at that 3-4 Number of the vessel, according to national station and the number of Type Master Header Records which have been filled number code. in for each observation type. 5-8 Station No. Consecutive numbers given for one year and one The Type Master Record serves as the header for the information relevant to ship (starting anew each year). the individual types of observations made. At present, five parameters are 9-12 included in this format, with code numbers as follows: Latitude Given to nearest minute. 13-17 Longitude Code No. Parameter Given to nearest minute. 18 NESW 01 Phytoplankton Primary Production Given according to the following code: 02 Chlorophyll ~igments 03 Phytoplankton 0-NandE 04 Zooplankton 1 N and W 05 Zoo benthos 2 S and E 3 S and W For each parameter measured at a station, one or more Type Master Records In this connection, the following rules are will be filled in, according to, e.g., whether one or more than one method observed: of measurement has been used. The first eleven columns are identical to Columns 1-8 and 19-21 of the BioMaster Record to provide for proper 0° Latitude is taken as N identification of the observation series (see Figure 2). 0° Longitude is taken as E 180° Longitude is taken as w. The Data Cycle Record provides the record for the individual data obtained 19-21 Year with respect to the relevant Type Vaster Record. For example, the Data Insert last three digits of the year. Cycle Record may contain information on the number of indi~idual organisms 22-23 Month and their biomass according to species for phytoplankton, zooplankton, and Number of the month. J anuary = 01, etc. zoobenthos. The first 11 columns are identical to Columns 1-8 and 19-21 24-25 Day of the BioMaster Record, while the next 4 columns serve to correlate the Day of the month. Data Cycle Record with the appropriate Type Master Record (see Figure 2). 26--27 Station time Starting time (to the nearest hour) of the The Plain Language Record may be used at any level in the format to insert hydrographic station in GMT. comments which are, e.g., relevant to the interpretation of the data. 28-29 Parameter ) Information on which parameters have been The first 11 columns of this record are identical to Columns 1-8 and 19-21 30-31 No. of Type Masters) on the BioMaster Record. The next 4 columns are used to identify the type measured at the station is inserted according of observation to which a particular Plain Language Record is relevant. 32-33 Parameter ) to the following code: 34-35 No. of Type Masters) 01 = Phytoplankton Primary Production 3· DErAILED DESCRIPI'ION 36--37 Parameter ) 02 = Chlorophyll ~!Pigments 38-39 No. of Type Masters) 03 = Phytoplankton 3.1. BioMaster Record 04 = Zooplankton 40-41 Parameter ) 05 = Zoobenthos The BioMaster Record is the file header for the full series of 42-43 No. of Type Masters) biological data obtained at one station. The BioMaster identifies 44-45 Parameter ) The number of Type Master Records in the file what type of information is contained in the file, i.e., which·parameters associated with each parameter is given in the 46--47 No. of Type Masters) subsequent two columns. have been measured and how m&~y Type Master Records have been prepared for each parameter. 48-72 Unassigned Reserved for definition later, as needed • One BioMaster Record should be filled out for each sampling station according . Insert blanks. to the scheme given below. The first 27 columns should be filled in with 73-77 ICODS Station No. information identical to that given in the corresponding Hydro Master Card Insert station number according to the for that station. For convenience, part of the scheme concerning these International Catalogue of Ocean Data Systems. first 27 columns is given here (see page 4); full descriptions can be 78 found in the Manual on ICES Oceanographic Punch Cards (Fourth Edition, Unassigned Reserved for possible future use for coding 1979). purposes. Insert blanks.

79-80 Bi·oMaster Record Insert the figure 31. Code -124- -125-

24-30 Latitude 3.2. Phytoplankton Primary Production ------24 N or S insert letter or 1 - North The number of Type Master Records to be filled out will depend 2 - South on the number of different methods or conditions used to measure 25-26 Degrees primary production at the station. If only one experimental method (incubator or in situ) is used ~~d if the same 14c counting method and incubation times 27-30 Minutes to the second decimal place are used for all samples, then only one Type Master Record should be completed for this parameter. However, if different experimental or counting methods ~2:~~-:0.:'~g~ tu~~ are used or different incubation times or durations are employed, separate 31 E or W insert letter or Type Master Records should be filled out for each different condition. The means of distinguishing among these variations are contained in Columns - East 14 and 15, with further specification in Columns 41-48, 49-51 and 52, as 2 - West applicable. 32-34 Degrees This Record should be filled out as follows: 35-38 Minutes to the second decimal place.

Column Item Code Description Column Item Code Descriution

1-2 Country Fill in same information as in Columns l-8 39-40 SB.mpling time Insert the time of sampling ln GMT to the 3-4 Ship on BioVBster Record. nearest hour. 5-8 Station No.~ 41-48 Incubation time Insert incubation time to the nearest minute 9-11 Year Insert last three digits of the year. in GMT:

12-13 Parameter InsertOl (code number for phytoplankton primary Cols. 41-44 beginning of incubation production). Cols. 45-48 end of incubation 49-51 Incubation tempe­ Insert temperature of incubation 1n °c to the 14 Method Insert code number of experimental method used, rature as follows: first decimal place. 52 Counting method 1 l - incubator Insert the method used to cournt~ 4c ac t"1 v1· ty: 2 - ---in situ 1 Liquid scintillation 2 - Geiger-MUller 15 Specification If measurements have been carried out using two 53 Attentuation or more incubation times or temperatures, or Insert the method used to determine the relative if different l4c counting methods have been used, determination irradiance at the sampling depths: assign a number for each variation, beginning method with 1. If no different conditions have been 1 - quanta meter used, insert a blank. 2 - photometer with green filter 3 - Secchi disc No. of depths Right justified. Insert the number of depths 16-17 54-72 Unassigned at which samples have been taken. Fill ·•i th blanks. 73 Irradiance unit 18-23 Depth to bottom Right justified. Insert depth to bottom in Insert the unit in which the irradiance 1n the meters, to the first decimal if possible or incubator is given: appropriate. If not, insert a blank in Column 23. 1 - 1018 quanta m-2 (400 1 700 nm) 2 - 10 8 quanta m-2 (350 - 3000 nm) 24-38 Detailed Coordinates These columns are available to specify the 3 - Joules m-2 s-1 (4oo 700 nm) (optional) coordinates of the sampling site in greater 4 - Joules m-2 s-1 (350 - 3000 nm) detail, if desired. Otherwise, the columns 74-77 Incubator ' should be filled with blanks. Right justified. Insert the mean irradiance in irradiance ~he ~~cubator during the experiment (or the mean If used, the columns should be filled as follows: (see next page) 1rraa1ance during the in situ experiment). 78 Unassigned Reserved for possible future use for coding purposes. Insert blank.

79-80 Type Master Insert the figure 32. Record Code -127- -126-

3.3. Chlorophyll a 3.2.2 ~~~~-Qz£~~-~~£~~~ uld be filled in for each sub-sample studied, One Data Cycle Record Sho as follows: The number of Type Y~ster Records to be filled out will depend Code Descrintion on the number of different methods or conditions used to Column determine chlorophyll ~ at the station. If only one method is used for all the Fill in same information as in Columns 1-15 of samples, then only one Type YillEter Record should be completed for this 1-15 the associated Primary Production Type Master parameter. However, if different solvents, filter types, or measurement Record. techniques (spectrophotometric or fluorometric) are used, then separate Type Master Records should be filled out for each different condition. The 16-17 Unassigned Fill with blanks. means of distinguishing among these variations are contained in Columns 14 and 15, with further specification in Columns 44-52, as applicable. Observation depth Right justified. Insert depth at which sample 18-23 was obtained in meters, to one decimal place if applicable. Insert a blank if no decimal place This record should be f-illed out as follows: used. Column Code Description Right justified. Insert the relative irradiance 24-27 Relative irradiance in percentage of the irradiance immediately below 1-8 Fill in same informab.on as in Columns l-8 on the water surface, to one decimal place if applic­ BioMaster Record. able. Insert a blank if no decimal place is used. 9-11 Year Insert last three digits of the year. 28-38 Unassigned Fill with blanks. No. of parallel Right justified. Insert total n~ber of parallel 12-13 Parameter Insert 02 (code for chlorophyll a ). 39-40 samples taken at the depth g1ven 1n Columns 18-23. samples Insert the number of the particular sample being 14 Method Insert oode number of measurement method, an 41-42 Sample No. reported on this record, beginning with 01. follows: Insert the total number of sub-samples obtained 1 - spectrophotometric 43 No. of sub-samples 2 - fluorometric from the sample. Insert the number of the particular sub-sample 44 Sub-sample No. 15 Specification If measurements have been carried out using biir being reported on this record, beginning with 1. or more types of filters, extraction solvent:', or other condition reported on the Type M::wtor Right justified. Insert the temper~tur~ - 45-49 Potential primary Record, assign a number for each variation production corrected maximum potential product1on 1n mgC m-3 hr-1 to the second decimal place. beginning with 1. If no different condi t:i.onn have been used, insert a blank. Right justified. Insert the day-integrated actual 50-54 Daily production per m3 primary production in mgC m-3 day-1 to the second 16-17 No. of depths Follow the instructions given for the idenL)N; i 18-23 Depth to bottom Icolumn numbers in the Phytoplankton Primal'.Y decimal place. 3 1 24-38 Detailed coordinates Production Type Master Record (Sec. 3. 2.1), Right justified. Insert •'n mgC m- hr- to the 55-59 Dark fixation (optional) second decimal place. . . mM dm-3 Sampling time 60-62 Carbon dioxide Insert concentrat1on 1n 39-40 Insert time of sampling in GMT to the m\M.'r'r'''' t; iv,HY, Reserved for future definition as necessary. 63-69 Unassigned· 41-43 Sample volume Right justified. Insert volume of sarnpJ o U1 r!Hr Fill with blanks. to one decimal place, Daily production Right justified. Insert the depth-integrated 70-74 actual primary production in mgC m-2 day- 1 to the Filter type Insert the code for the type of filtH per m2 44 first decimal place. as follows: l - Whatman GF/F or GF/C 75-77 Part counted Right justified. Insert as per cent, with no (optional) decimal place. 45-46 Filter diameter Insert the active filter diameter in r1d 11 78 Unassigned Reserved for possible future use for coding purposes. Insert blanks. 47 Solvent Insert the code number for the extrrl.GLi.i.Al used, as follows: 79-80 Data Cycle Insert the figure 33. Record Code 1 - 9o% acetone. -129- -128-

Code Description 3.4. Phytoplankton Column 3 48-50 Extraction volume Insert in em • Normally only one Type Master Record for phytoplankton wi.ll ],,, 51-52 Cuvette size Insert ~n mm. needed for each station. If, however, different preservati.Vhfl are used, a separate Type YJaster Record should be completed for each bnd cd Calculation method Insert the method used for calculation of the preservative. A description of the record is as follows: 53 concentration of chlorophyll-a according to the following code: Column Item Code Description 1 Jeffrey & Humphrey equation 2 - Lorenzen equation l-8 Fill in same information as in Columns l-8 •>~< BioYJaster Record. Reserved for future definition. Fill with 54-17 Unassigned blanks. 9-ll Year Insert last three digits of the year. 12-13 Parameter Insert 03 (code number for phytoplankton). Unassigned Reserved for possible future use for coding purposes. Insert blank. 14 Method If agreed method is used, leave blank. If il!d, Insert the figure 32. insert 9 and describe differences on Plain Lc"'l' '''.!'' 79-80 Type Master Record. Code 15 Specification Insert blank, unless two or more preservatl v•·•tt kF' been used.

16-17 No. of depths ~ Follow the instructions given for the irl••>liiu•i 3.3.2. ~ata_Qyc~~-~~£E~ 18-23 Depth to bottom column numbers in the Phytoplankton Prin~•rv 24-38 Detailed coordina- Production Type YJaster Record (Sec. 3.2.1).. One Data Cycle Record should be filled in for each sub-sample, giving tes (optional) ) the information as follows: 39-40 Sampling time ) Code Description Column Item 41 Preservative Insert according to the following code: Fill in same information as in Columns l-15 of l-15 l - Lugol + acetic acid the associated Chlorophyll ~ Type Master Record. 2 - Lugol 3 - 2% formaldehyde 16-17 Unassigned Fill with blanks. 4 - "Keefe" Follow the instructions given for the identical 18-23 Observation depth Unassigned Reserved for future definition, as needed, !Vdi column numbers in the Phytoplankton Primary 42-77 24-38 Unassigned with blanks. 39-40 No. of parallel Production Data Cycle Record (Sec. 3.2.2). samples 78 Unassigned Reserv'ed for possible future use for codil•lK 41-42 Sample No. purposes. Insert blank. 43 No. of sub-samples 44 Sub-sample No. 79-80 Type Master Record Insert the figure 32. Right justified. Insert in mg m-3 to one Code 45-47 Chlorophyll ~ decimal place. 3 Right justified. Insert in mg m- to one decimal 48-50 Phaeopigment place. ' One Data Cycle Record should be filled out for each fl)HAid v•t• Reserved for future definition as needed. Fill found in each counting aliquot, which is a known fracU.on 51-77 Unassigned with blanks. sub-sample divided for purpose13 of counting the organisms. The reoo:Ni be completed as follows: 78 Unassigned Reserved for possible future use for coding purposes. Insert blanks.

79-80 Data Cycle Record Insert the figure 33· Code '""ll!l,,------130- -131-

Column Column Code Description Code Description 59-63 C0ll volume Insert appropriate cell volume for the units of 1-15 Fill in same information as in Columns l-15 of the associated Phytoplankton Type Master Record. organisms counted in ~rn3 using scientific notation according to the following scheme: Fill with blanks. 16-17 Unassigned Col. 59 integer Cols. 60-62 decimal places (insert blanks in columns 18-2) Observation depth Right justified. Insert sampling depth in meters, with a blank in Column 23 (reserved for decimal not otherwise filled) Col. 63 exponent place). If an L~tegrated sample is taken over a Example: range of depths, give minimum and maximum depths cell volume is 435 according to the instructions given for the identic:Ji ~3 change to scientific notation: 4.35 x 102 column numbers in the Zooplankton Data Cycle insert in columns: Record (sec. 3.5.2). 4 3 5 b 2

24 Species code Insert identification number for species code used, 59 60 61 62 63 identification as follows: l - USNODC Taxonomic Code a blank is inserted in Col. 62 as the decimal place is not otherwise filled. 2 - Plain language abbreviations of Latin name (6 cols. for genus + 6 cols. for species). 64-68 Plasma volume Insert plasma volume in ~3 using scientific notation and the same scheme as for cell vo1urno, 3 - RUBIN Code List PL Phytoplankton, limnic. above. 25-36 Species Left justified. Insert code number of species or Col. 64 integer plain language abbreviation. Blanks should be Cols. 65-67 decimal places (insert blanko Jn '"' 1UiiHl!> inserted in columns not used. not otherwise filled) Col. 68 exponent 37-38 Identifiers Insert code for appropriate identifiersr e.g., See example for cell volume. developmental stage, sex, size, etc. L Code to be developed later._7 69-73 Carbon content Right justified. Insert in picograms.

39-40 No. of parallel ) Follow the instructions given for the identical 74-77 Unassigned Reserved for future definition as necesoal',Y, ll column numbers in the Phytoplankton Primary with blanks • samples 41-42 Sample No. l Production Data Cycle Record (Sec. 3.2.2). 43 No. of sub-samples 78 Unassigned Reserved for possible future use for codJ rltt !H 44 Sub-sample No. Insert blank.

45 No. of counting Insert the number of aliquots into which the 79-80 Data Cycle Record Insert the figure 33. aliquots sample was divided for counting purposes. Code

46 Aliquot No. Insert the number of the particular counting aliquot for which results are being reported on 3.5. Mesozooplankton this record, beginning with 1.

47-49 Magnification Insert amount of total magnification used in counting the aliquot. The number of Type Master Records to be filled in wiJl number of different methods used to obtain and analyo;, 50-54 Coefficient Right justified. Insert the appropriate coef­ plankton samples at the station. If only one method is used for ttll ficient for the species reported according to the only one Type Master Record should be completed for this parameter, magnification used and other relevant factors, if more than one, mesh size sampler is used to obtain the samples, as is needed to express the number of units of the then separate Type Master Records should be filled in for each organisms per dm3. condition. NOTE: At present,' this record is intended for data oll vertical hauls only. Revisions must be made before data obtained uxd 55-58 No. of counting Right justified. Insert the number of counting oblique hauls can be recorded for this parameter. units units of organisms (as defined for each species - cells, clusters, etc.) observed for the species being reported. -132- -133-

This record should be completed as follows: Code Descrintion One Data Cycle Record should be filled out for each species found in Column each sub-sample counted. The record should be completed as follows: Fill in same information as in Cols. 1-8 on the 1-8 BioY~ster Record. Column Code Description

Insert last three digits of the year. 9-ll Year 1-15 Fill in same information as in Cols. l-15 of the associated Zooplankton Type Master Record. 12-13 Parameter Insert 04 (code for zooplankton). 1&--17 Haul No. Right justified. Insert the number of the haul Insert code number of type of sample obtained, as 14 Method from which the sample being recorded was taken. follows: 18-20 Maximum depth of haul Right justified. Insert maximum depth in meters. 1 - net sample 2 - water volume sample 21-23 Minimum depth of haul Right justified. Insert minimum depth in meters. Specification If samples have been obtained using more than one. 15 type of sampler or if they have been preserved us1ng 24 Species code identi­ Insert identification number for species code used, fication as follows: more than one type of preservative, for exampl:, assign a number for each variation begi~n~ w1th 1. 1 - USNODC Taxonomic Code. If no different conditions have been usee, 1nsert 2 - Plain language abbreviation of Latin name (6 a blank. colwms for genus + 6 columns for species). 3 - RUBIN Code List 01 Baltic Invertebrates. 1&--17 No. of hauls Right justified. Insert number of hauls taken at 25-36 Species Left justified. Insert code number of species or station. plain language abbreviation. Blanks should be inserted in columns not used. lB-23 Depth to bottom ~ Follow. the instructions given for the identical column numbers in the Phytoplankton 24-38 Detailed coordinates 37 Unassigned Insert a blank. (optional) Primary Production Type Master Record (Sec. 3.2.1). Insert the time of sampling in· GMT to the nearest 38 Developmental Insert code for developmental stage and s·ex: 39-40 Sampling time stage and sex hour. 1 - copepodite stage I-III 2 - copepodite stage IV-V Insert code number for sampler used, as follows: 41-42 Sampler 3 - adult female 4 - adult male Ol - WP-2 net 2 4)-47 A:rea of sampler Right justified. Insert in em • 39-40 No. of parallel~ Follow the instructions given for the identical opening samples column numbers in the Phytoplankton Primary 41-42 Sample No. Production Data Cycle Record (Sec. 3.2.2). 48-50 Sampler volume Right justified. Insert in dm3. 43 No. of counted sub­ Insert the total number of sub-samples in which 51-53 Mesh size Insert mesh size of net used to obtain sample in pro, samples zooplankton have been counted.

54 Preservative Insert code, as follows: 44 Sub-sample No. Insert the number of the particular sub-sample being reported on this record, beginning with 1. 1 - 4% formaldehyde buffered with borax 2 - l:l ~ ethanol : 4% formaldehyde 45-47 Portion counted Insert the denominator of the fraction into which the sample has been divided, e.g., 1/64 is recorded 55 Wet weight deter­ Insert code, as follows: as 064; 1/8 is recorded as 008. mination method 1 - individual volume (fresh weight) 2 - weighing (formalin weight) 48-51 No. of individuals Right justified. Insert the total number of 3 - displacement (formalin weight) counted individual organisms of the species which were counted in the sub-sample. Reserved for definition later, as needed. Fill 5&--77 Unassigned Biomass - wet weight Insert the wet weight of the individuals counted with blanks. 52-56 in ~g. Scientific notation should be used and the number recorded as follows: 78 Unassigned Reserved for possible future use for coding purposurr Insert blank. Col. 52 integer Cola. 53-54 decimal places Insert 32. 79-80 Type Master Record Cols. 55-56 exponent r.nclP. -135- -134-

Column Code Description Code Description Column 67-71 Carbon content Insert carbon content for the individuals counted Example 1: in ~· Scientific notation should be used and the the wet weight is 56.8 mg number recorded as follows: convert to ~g = 56800 ~g 4 Col. 67 integer express in scientific notation: 5.68 x 10 ~g Cols. 68-69 decimal places record as follows: Cols. 70-71 exponent, 5 6 8 0 4 72 Unassigned Insert blank , 52 53 54 55 56 73-75 Filtration For use only when flow meters are employed on insert zero in Col. 55 to fill space. efficiency sampling nets. Right justified. Insert filtration efficiency in per cent as the ratio between the Example 2: amount of water passing through the net and the the wet weight is 0.093 ~ -2 amount of water flowing outside the net. express in scientific notation: 9.3 x 10 ~g record as follows: 76-77 Wire angle If wire during sampling was vertical, insert 00. If wire was not completely vertical, insert 9 3 b 2 approximate angle by which it deviated from the vertical, e.g., the figure 10 would indicate a 52 53 54 55 56 10° deviation from the vertical. insert a blank in Col. 54 and a minus sign in insert the 78 Unassigned Reserved for possible future use for coding purposes, Col. 55 (if minus sign cannot be used Insert blank. digit 8). 79-80 Data Cycle Record Insert the figure 33. Example ): Code the wet weight is 3.42 ~g record as 3.6. Zoobenthos 3 4 2 0 0 52 53 54 55 56 insert zeros in the exponent columns. The number of Type Master Records to be filled in will depend on the number of different methods used to obtain and analyse the lnBert the dry weight of the individuals counted in zoobenthos samples. If only one method is used, only one Type Master Record should 57-61 Biomass - dry weight ~g. Scientific notation should be used and the be completed, However, if different methods are used to obtain the samples number recorded as follows: different mesh sizes or preservatives are used to treat the sample, separate Type Master Records should be filled in for each different condition. Col. 57 integer Cols. 58-59 decimal places This record should be completed as follows: Cols. 60-61 exponent See examples in wet weight biomass, above. Column Code Description

Insert ash weight for the individuals counted in ~g, l-8 Fill in same information as in Cole. 1-8 on the 62-66 Ash weight Scientific notation should be used snd the number EioMaster Record. recorded as follows: 9-11 Year Insert last three digits of the year, Col. 62 integer Cols. 63-64 decimal places 12-13 Parameter Insert 05 (code for Zoobenthos). Cols. 65-66 exponent See examples in wet weight biomass, above. 14 Method Insert the means of obtaining the sample according to the following code:

1 - quantitative ~ab sampling 2 - diving (scuba) 3 - dredge sampling -137- -136-

Code Description Column Item Code Description Column 16-17 Unassigned Insert blanks. Specification If samples have been obtained using more than one 15 type of sampler or if more than one size of mesh. or type of preservative has been used on the var1ous 18-23 Observation depth Right justified. Insert sampling depth in meters tr samples, assign a number for each variation beginniDf, one decimal place if possible. If not possible, with 1. If no different conditions have been used, insert a blank in Col. 23. insert a blank. 24 Species identifi­ Insert identification number for species code used, Fill with blanks. cation code as follows: 16-23 Unassigned 1 - USNODC Taxonomic Co

*) If bottom sediments are sampled the Sample Description Form given on page 38 of this Annex may be used. 10 483184A -138- -139-

Column ~ Code Description Column Item Code Description

62-66 Ash weight Insert the ash weight of the individuals counted in 16-77 Comments Fil~ in with comments in the English language, If mg. Scientific notation should be used and the l.t ls necessary to identify the record with a number recorded as follows: particular sample or sub-sample, the numbers of Col. 62 integer these should be inserted in Cola. 41-42 and 44, Cols. 63-64 decimal places respectively. If further identification to a Cols. 65-66 exponent particular species is necessary, the species code should be inserted in Cols. 25-36. See examples mentioned under formalin wet weight above. 78 Unassigned Reserved for possible future use for coding purposes. 67-71 Carbon content Insert the carbon content of the individuals counted in mg. Scientific notation should be used 79-80 Plain Language Record Insert the figure 99. and the number recorded as follows: Code Col. 67 integer Cols. 68-69 decimal places 4. GENERAL REMARKS FOR ALL RECORDS Cols. 70-71 exponent · Except where stated otherwise in the above sections, whenever an element is Unassigned Reserved for definition later, as needed. Fill with 72-77 missing this should be indicated by filling the whole field for that element blanks. with blanks.

78 Unassigned Reserved for possible future use for coding purposes. Insert blank. All columns must be filled for every observation according to the scheme that columns for missing digits before a decimal point should be filled with zeros while columns for missing digits after a decimal point should be filled with ' 79-80 Data Cycle Record Insert the figure 33· blanks. For example: Code a depth to bottom of 85m is recorded as 00085 blank 3, 7 • Plain Language Record a zooplankton wet weight biomass of 3.9 x 102 ~g is recorded as 39 blank 02.

This record can be used to insert explanatory notes and other information Many of the code listings for methods, samplers and preservatives are incomplete. relevant to a proper interpretation of the data. The record may be used Suggestions for additional items for these codes should be mailed to the at any level in the format and as many record~ may.be used as are needed to Environment Officer, International Council for the Exploration of the Sea, report the desired information. In order to ~dent~fy the re?ord. properlJ:' and. , Palffigade 2-4, 1261 Copenhagen K, Denmark (after 1 January 1980). place it in its appropriate position in the file, the follow~ng l.nformabon snouh be provided on each Plain Language Record:

Column Code Description

1-8 Insert the information contained in Cols. 1-8 of the associated BioMaster Record.

9-11 Year Insert the last three digits of the year.

12-13 Parameter Insert the code number of the parameter to which the record is relevant. If the record is associated with the BioMaster level, insert 00.

14 Method ) Insert the appropriate figures from the relevant 15 Specification) Type Master Records. -141- -140- Appendl'X · 1 ,_, IOC COUNTRY CODES FOR ICES MEMBER COUNTRIES Country ot:J N ""00 "" f.,< Ship " [,r ~"': '-""'1-'<+ Country 'E) ;::;; "'1 Parameter _, Station No. l3elgium 11 e "' ""'() 0 It; "' '1 Canada 18 No. of Type Masters "" 0 ~ 1!=: 1----1--- -}--1-'4------Denmark 26 t Parameter ~ Finland 34 ~ No. of Type Masters ~ t" France 35 !"': 0 0 ~ Jl 96 ~ 1-' .... German Democratic Republic --- - -f--~~~------55 11' Germany, Federal Republic of 06 cg - """' 46 ~ NSEW Iceland t;(j Ireland 45 ~ Year Netherlands 64 ~· N 58 ~ N Norway i

~ Record Code 0 ' ' "' -142- -143-

,.., CountrJ .... Country N N "' Ship tv' Ship ""- ""'" ~ V> ~ "' Station No. ~ _, Station No. ft I-' Sample No. "' Ol f; 0> e '-D No. of sub-samples '-D .... e .... 0 Year 0 Year f-+------~ Incubation time rt Sub-sample No. "'" ...... fs; N .... ~ Parameter ~ Potential pri~ary rv Parameter .... product:i.o:1. K:;; ~ ...... ~ (mge m-3 hr-1) It: Method Method ~ ~ !t: i:;; Specification Specification ~ ~ ~ I. (°C ...... ~ Incubation temperature No • of depths fcg ...... "fi "'.... "'(Unassigned).... f;::; Daily oroduction .... per m3 .. 1-' ~ Counting method Ol 0> ...... ~ (mge m-3 day-1) ~ Attenuation determ. meth. ~ ~ N ~ N 0 Depth to bottom t;;; 0 Observation depth N N .... 1-' N ~ N ...... N . '(j; ...... ~ Dark fixation "'N ...... ';\ (mge m-3 hr-1) '"' - - --"- ~!en.~- \£ ~ ~ 0 ~ Relative ~ ~ 8 N ....<+ N irradiance 1-- ~-- - ~~-- ...... (Unassigned) - N <+ 8 . ."' a--,.., ~ Carbon dioxide f--l m.... ~ N "' mM dm-3 N (;) .. " t:J m co ...... N "<+ p; .... ~ fc; ".... ~ ic'; ~"0 p. .)12 (Unassigned) 'd M-D ;:: -~-- -+-- .::::~~."' -- ..-.o 0 0 ~ ~ ~ ~ p. m 0 !:""' ...... " "' (Unassigned) 0 " ~ ~" ~ ~ Y' (Jq".... "<+ ~--- Ol ~ 1------<+ " "'0> ~ ~ ~ ""' ~ ...... ~ - f(3 ic;', Daily oroduc~ion ~ f;:; per ,2· ~ lc; 2 1;:; (mgC o- C.ay-1 ) lc; ~ Irradiance unit No. of parallel ~ Sa:opling time f--' ~ ...... sarople• ~ It -~-t;;: Po . ~ I~cubator i:::-:-adiance p !(;; Part counted (%) !(;; ' r: f-.< 1;3 (Unassigned) ~ (Unassigned) Record code ~ fZ Record code 8 0> 0 -144-

···r '""""··v··""""~• ~-"" >-' Country ~· Country ~"" N M>--~ "' -- ~--"~ r-~ ~·-·· ~ Ship lv- Ship fl>.. "'(l) r-- --- f;: tv' "'"tv' It m 3 "' Station No. " Sample No. Station No. Sample volume (ctm ) r fl;; r ...... ()) .. ~ \:;; No. of sub-samples OJ ~ t"' Filter type >-' . Sub-sample No. It: 0 Year It: b Year >-' >-' >-' ~ Filter diameter ~ Chlorophyll >-' >-' ~ >-' ~ Parameter . . f0, "' ...... (;ng m-3) N Parameter ~ Solvent >-' ~ 0 ~ 0: N !g t:' Method ~ Method 3 Phaeopig:nent fl;: Extraction vol=e (cm ) Specification f:i; ~ ...... (mg m-3) ~ Specification ~ >-' ~· 1-' "'I-' No. of depths "' (Unassigned) ~ ~ Cuvette size (mm) t:::' ~ >-' ()) >-' ~ ~ OJ Calculation method ~ ~ ~ ~ ~ "'0 Depth to bottom ~ "'0 Observation depth N ~ "'>-' f:;i >-' ~ ...... "!"' ~ ...... "'ry ~ '-""' ~ ~ tcG "' UJZ ~ - t-- -- r-- - f'= -- ~ li ~ 0 1:" ~ ~ 8 N - - - "'t---"' "'"",... "'0 - r- N "' >-' c"" >-' "' r P< "' (-'"' N ()) t1 "' ...... - "' "' c ~ "" ::;;' "' "' ,_.. (\JP.assigned) ~ " ~>-'"' p "' :co 0 Z' rc; "',_.. (Unassigned) '§ !;.::: "'"' "'"'"" 3;' i;:; ~ - r---- 0 ()0 -- r-- -- r- -- ·- g; P< ID" H0 I'Ki f-'P< "' ~ 0 ~,... "' "' ~ t" ~ 0 " ~ ()) !:':;~ ____ "'"" ()) ""' ----r-- -- C'·" m "' fi: -- "'2" ~ P< ~ ~ ~ lc3 ...... rc;_ 1-3 to:, r;::; - ~ 1;::) ~ r;:;; r;:; ~ ~ ... ~ f::D Sa.mn ling ti;:ce f:;! ~ No. of parall·•l ~;::· 13 - samples fi> f;: ' lt ~ ~ to; 1;:;:' ' b t-!"' a; Unassigned a; (\Jnassigned) :;; ::;3 :fRecord Code Record code C) 0 0 Phytoplankton (Parameter No. 03) Data Cycle Tiecord, Code 33

0 -.-<" +' ~ "'0 ~ ....·.-< rl ·.-< "' .a +' ~ +' Q) " .--<"' Q) -.-< Q) "' "" .--< 0 ~ Q) .-1 ·.-I" "" 0 ,_, +' Q) 0 0 "' z "" ·.-1" ""0 ·~ :J"' "'0 e, +' ·.-1"' +'"' ·.-1 ..... '·-~ '"'' i7 0 .... __"'Q) , __"'Q) , ·.-I 4< +' ·.-1" " 0 ·.-1 "' "'t; <' 0 p, +' ~ .a"" 0 "' 0 0 § ~ +' ~ "' U<" :2 p p"' "' "'p, "' "' ~ 0 iii ~ ~ ~ ~ "' 0 U)"' "'H ~ l 2 _3 4 15 6 7 8 9 10 ll 12 l) 14 1'i"' 16 17 18 19 20 21 22· 21 24 25 26 27 28 29 30"' 31 22 33 34 35 JG 17 ';8 39 40

. I - - - - I

I ...,...

+' "'0

g. ~' - ·.-I "' "' rl § ~ rl"' "' ~ "'s -.3- "' . il' il' "'El +' ~"' 0 ·.-1 .... "" 3 ~ z +' 0 +' "' Q) ~ "'I . ·.-1" +' @ +'" ''d Q) . p § 0 +' § "' rl 0 rl 0 z 0 @ 0 0 "" 0 "' 0 " "' "' "" z 0 "' -.-1" 0 .--< ,. 0 f;, §, 0 ""' +' ..... "'0 0 •.-I ...... "'~ .... 0 ...... ,. .--<"' 0 0 .'H 0 ~ "0 "' "' f-l '"I & .... .--< p " "' "'0 "' p . ·.-1 . .-1 "'ol ~ a 0 ~ 0 0 .-1 "'0 0 U> ~ p leo U) z a z ... 1 0 z 0 ~·P-< 0 ~ ~ "' 141 42 I 4' 144 145 146 ! 47 48 45 I 50 51 52 53 54 I 55 56 57 58 I 59: 60 61 621 Gl 6~: 65 66 67:68 69~0~1 ~2 _I 7 4 75 76 7 78 79"' 80 I . I . I I . ! i l I ! j _l ----- . -- -···

Phytoplankton (Parameter No. 03) Type Master Record, Code 32

El 0 Detailed coordinates ., +' 0 .a +' (optional) -~"' ·.-1 +' 0 +' " p 0 ,_, +' z () "' 0 ~ +' ....."' "' +' ..... "' "" Latitude Longitude .--< i7 0 ...... p, +' ·.-I 0 .... 0 .a " ~"' "".a () +' § +' ~ +' . Po "' ~ p, 0 Q) I . E 0 ! ' ~ 0 :2 +'"' ;.:: Ul"' A W I Ul 0 >-<"' "' z "' "' "' 28:29_30 "\1 32 33 34! 32_38:37 38 39 40 1 2fl:41s 6 1 8\9 10 n\12 l3jl4jl5jl6 i7jl8 12 20 21 22 :z -,

' 1I I _]_' I I I I I _L I I _]_' ..\0 rl I

'CJ "' > 0 ....."' "' ""0 +' fi, ·.-1 ,_, Unassigned I "'t; ""0 "' 0 ~"' "' p "' &-:"' "' ]8 79 80. i4l .12 ,j_~ .1.1 45 46 47 .18 .19 50 51 52 53 'J4 55 'i6 'i7 'i8 59 60 61 62 6) 64 6'i 66 67 68 69 70 71 72 7"1 74 7'i 76 "71 -148-

>-' >-' N Country Country tv< N p. Ship ~ Ship (,. ~ f-1' It: No. If': Sampler 1-<"' Station No. "' Station No. N CD 1-' counted sub-samples CD )-!: ~ >-' Sub-sample No. f-a 0 Year >-' 0 Year ~ 2 >-' ~Area of sampler opening ( cm ) >-' ,...,>-' >-' counted ~-·~•W'>>' N >-' ~ Parameter N >-' Parameter \:"': K;: Method >-' Method ~ t: .. ;:;; Specification f:i; Sampler volume (dm3) o. of individuals ~ Specification >-' ~-·"'" res No. of hauls >-' "'>-' Haul No. t;:: "'>-' >-' ~Mesh size (mm) CD >-' ' CD I I ~ p Maxi= depth of haul (m) N ~ ~ Preservative 0 N ' Depth to bottom (~) 0 N -·" f-c l-et weight detemination method >-' N I >-' ' N f0 N N Mi niillUlJl depth of hau) ( .·,J J ...... N "' N I -" I...... ~ UlZ Species code ident.i.ft'ii'i~ H I cCD ~· I--f--f---~ f---- ~ r~ I fJi I 0 (J ~ N t-< N I 0 I "' --1---f---I--~ ~---- <+ cr-. \'3 ">-'• l "'>-' <+c \'3 N p, (J) t:J N f N .. . . - . . "'r-.: CD "' . . .. " "<1' I ::;; ~ >-'· "' (25 i fcj ~>-'" I '00 p," sh weight(eg) fcj Species 122 <+ >:t':l >-'· () 0 0 i f3i q - ~-'--- --~ -- ~ cr-. " 0 ~ ">-'P. 'i )r:; I "Ol ~>-'· "' Ol" 0 t-< ,... ~ 0 ~ l ~ <+ ~ cr-. '§ CD ,.... "Ol p, 1------" ~ I " - ~-~ "'2' p, Carbon content (eg) I ~ ~ I f--c ~ " 0 I 0 . . fa:, ...... ~ - •.. { I ~ ~ \Jnassign.ed I' f;l Ia; "' (Unassigned) l' CD Devel01ocent ,,,,., .. ·, i I p ~ Sampling time ~ No. of parall<~l (ii,jt.!(j I ~ Filtrati~n efficiency (%) I I ~ ~ I I b ' fa; Wire angle [:::3 (Unassigned) fa; \U~.assigned

~ Record code D fRecord code :;g 0 -150-

•'-' '- ~""•'»-'•••w~~~··- !-··· ...... oun t:-y t:JN Country ('.) ""W'- ---- ·····<'~' ••w"" "-~ . ~---~-~--- "'<+0 0 "' Jl ''"<) ' ~ Ship 'JUp " cr' ..__ ~:: O:o> ~ __ .. '< "c+ .1• t--n f.:; >-'00 "" It Station No. Sample No. [F., Sampler ",;:; ;:station No. "' "' f-l"" "'~ (D "' ., "' CD No. ~ :"" of sub-samples g ~ t--o lt Sub-sample No. "' 8.~ 2 ..... :=)!rear - "'M- ~fi,. Sampler axea (em ) 0 Yeax om ...... 0 ~ p; ..... ~ ..... p, Paxt counted " 0 ~ ..... ~ ..... V' N Paxameter "'!parameter "'~ :::; ..... ~ ::;; "' "' ~ ~ Method It;: Method ~ Mesh size ( fllil) ~ (!; l--' Specification ~ Specification No. of individuals ~ ..... ~ ..... ~Preservative "" ~ ; (Unassigned) ~ q ~ ..... ~ ...... (D "' ~ ~- ~ ~ ...."' ~ ~ "' --- ~- For:nalin wet weight (mg) 0 '§ - -- "' "' ~ ~ bbserva tion depth (m) f::; N "' ..... "' ..... ~ ~ N N"' ...... ~ f::i (;; ...... ~ (;; ~ q (D ~ ~ cnz ~ ~pecies identification code " - dry 00 1---1--- Formalin weight (mg) f:s "'00 - ~- pi .... ~ 0 3' 3' '§ t:-< "' f--· ~ -- ..... "" ~ "' "'....c+ "" ~ "' ~ c+ ~ t:J N N ~ p, ...... "" (D (D . . . "' c+ ...... - " " ~ ~ "'>"· ~ ~ ~ ..... ~ Ash weight (mg) ~pecies ~ }() 0 " 1---1-- -- r= 15 <+ ""'"'>"· 0 0i ~ kt>J 0 0 ~ --f---f-- f---1-- ~ ,-- " 0 "" "" li; "'f-'P, ~ "" ~ "" ~>"· 0 t:-< f2i 0 ...... ~ ~ 0'> ~ ~ c+ (D ~ ~" " ~ f---:-· tt <+ "' G;' --- ~ Caxbon content (mg) p, f------~ t:;; ~ ~ " fc; ...... ~ - f;j ~ f,:: ~ ~ ~ dentifiers ~ ~ a: ~ ~ ~ Bottom type fil Sampling time lo. of parallel samples :;;: :;;: It It (Unassigned) mo-'"" ::;; ::;; ' ;;; ;;; .._, :::3 ' fa; Unassigned fa; (Unassigned)

f:z Record code Pi Record C) C) code 0 0 08 6L 8L iLL 9L SL VL U i'.L 1L OL 69 89 L9 99 '9 1>9 >9 Z9 19 09 6S 9<; Ls 9S SS VS loS <;<, 1S OS bV 9V LV 9V £1> VV >V ZV TV q 0"' ~ "'0 " >'·"' "'0 'tl 0 "' "' "'

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Coun p.,,.- No Unaaa;gnood - Stolp St•llon No lal.tuda longUud• NS Y.. r Station Par1- No Para. No F'au- No No P•••- ·~ EW """" "'' malar of Typo<~ malar ot Type ..,., .. of Typo matar or Typo mala< or Typ. c- "' o : I """' Mlt..,le<"S M~,l~r , ~o,~,,,~, -~ "'"~ STATION 1 2 IJ • 5 6 7 6 19 10i1! 12 13 14• 15; i: 16 '17 18 19 20 21 22 2J 24 25 26 27 28 29 30 31 32 3 435 b6 37 3839 0 41 2 43 44 4 464 7 48 49 50 51 52 53 54 55 56 57 58 59 60 61 ' L J I I I I l I I I i I I I I I I I I I I I I _I I I I I I I I I I I I I I I I I ' ' . I I ! ! I I I ; I I I t I I I I I I I I I I I I I I I I I I o I t I t t I I I I ' ' I I I I I I i I I I i I I I I I I I I I I I I I I I I I I I I I I I I I ' ' ' ' ' I I I I I I j I I I l I I I I I I I I I i I I I I I I I o I I I I I I I I I I I 1L! ' I I I I I I j I I I : I I I I I I I I I I I I I I I I I I I I I I I I I I I I I ~ ' 3 I I I I I I I I i I I I I I I I I I I I I I I I I I I I I I I I I I I I I I ,I - l l ' ' '· 9 L . .J I I I i L I I i I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 3.1 i 1-L I ' ' 3.1 _L I I I I I ; I I I t I I I I I I I I I I I I I I I I I I I I I I I I I I I I I L_J_L_ ' ' 1 -J I I I I l I I I l I I I I I I I I I I I I I I I I I I I I I I I I I I I I I ' ' I I I I I I I I I t I I I I I I I I I I I I I I I I I l I I I I I I I I I I I I I I f-' II~ lJ1 ' ' 31 I _[_ I I I I l I. I I l I I I I I I I I I _[_ I I I I I I I I I ! I I I I I I I ! I ~ lJ1 I ' ' I. I I I I I I l I I I f I I I I I I I I I I I I I I I I I I I I I I I I I I I I ~! ' ' 1 I 1 I I I i I I I j I I I I _L I I I I I I I I I I I I I I I I I o I o < < I I ~ ' ' I I J j J _Lj_L _!_L_LL I I _L I I I _[_ _L _L I I I I I I I < I o I I I I I I I I I ~ ' _j __LL _L I I I _[_ _L I I I I I I I I I -1 I I I I I I I I 311' I I I I L 1-L! I II II .. (_L _L ' 31 I I I I I ILL _Ll l I I I I I I I I I I I I I I I I I I I < t < I I I I I I I ':X ' ' I I I I I I f I I I l i - I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I ~ ' ' 1 1 I I I I l I I I l I I I ! I I I I I I I I I I I I I I I I I I I I I I I I I 3Ll_ ' I I I I I I i_t_ I I ; I I I I I I I I I t I I I I I I I I I I I I I I I I I t I ~ ' ' I I I I I f I I I l I I I I I I I I I I I I I I I I I I I I I I I I < ~ ' ' ' ' I I I I J I i I I I j I I I I I I _[_ I I I I I I I I I I I I I I I I I I I I I I ~ ' ' I I I I I I i I I I t I I I I I I I I I I I I I I I I I I I I I I I I I I I I I ~ ' ' I I I I I I l I I I l I I I I I ' I I I I I I I I I I < I I I I < I I I I I I I ~ ' ' 3,1 I I I I I I I' I I I I' I I I I I I I t I I I I I I I I ! f I I I l I I I I I I I Coun. Shop St~>t••n Nc. Year P11ra- j gl STATION: PHYTOPLANKTON PRIMARY PRODUCTION met•• ~ ~ "' ; I COUNTRY 1 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 INSTITUTION· I I I I I I 1 011 l TYPE MASTER RECORD

No. ol! Oeptl> to bottom Deta•led Coordonates \Optoonat) Samo! lnc>.Jbat10n ,,..,.,\GMT) Incubation., • lJnns•gnod '2 Incubator\ I'~""' dept~ Lahlude l.>fl9•h.>do <"9 tempr!la· ~~ ~ ~ lry Dally Dark llxatoon Port Relative Carbon! Ut>l'''''gnod Daily '~""' "9"'" depth parallel sample -g "' production production 3 conted 'rradianc No. dioxici.4 produ2tior """" 0 -g C/mg m·3h;1 C/mg nB~~Y~ Cf mg ,n3 hr-1 3 ,,, z ~ ":" dm - 16 17 1819 2021 22 23 24 25 26 27 28 29 30 31 32 33 34 3536 37 38 394·- 41 42 4344 4546 47 48 49 50 51 52 53 54 55 5657 58 59 6061 62 636465 66 67 6869 1JH 1~ 7374 ~5 76 7 7 798C Remarks : I I I I ; I I I I _LLLL _L_LLl I I ; I _II I I I I I I I I I I I I 313 I II II i I I I : :-: ~~: :~:~ I I I j I I I ; I I I ; I . I I I I I I I I I I I 313 I , I I II I j I I I I I I I I I I I I j _j I IIU I I ; I LLL ~L _LLL I I I I I I I I 3,3 ,_.I I I I I I ; I I I I I I I I I I I I .. - I I I l I I I ; I I I l L L l l_l Ll_l I I L 1 J I I 3,3 .Lt.L ()1 I I I II l I I I I I I I I I I I LLr I I I I l_L_ _j_j__l __ L __l_Li I I I I I I I I I I I I I I I 3,3 "'I I I I I I j I I I I I I I I LLLL I I "u_ _L_Ll_L J I l I I L J__L I I I I I I I I I l 3,3 ' I I I I I l I I I _I I I I I I I I I I I j I I l I I I l I L I l I . I I l l I I I I I I I I I 313 ' I I I I I l I I I I I I LLLLl_LL .. J I !IlL _LLLL I I l I -I I I I I I I I I I I I I I 3,3

I I I I I I j I I I IIIJ.!III_j_j I _L _ .U j I l I _j_L _LL_LL r_l I I I I I I I I I I I 1 j 3,3 I I I _I I I l I I j I 313 ~ I I I I I I I I I I I I I I I I I l I I I l I I I I I I I I I I I I I I I I I I _I I I l I I I I I I I I I I I I I I I Ll j I I I l I I I l I I I I I II I I I I I I I I 313 ~ I I II I l I I I I I I I I I I I I I L I I I I I I I l I. I I i I I I I I I I I I I I I I I I 3,3 iII i I I .l I I I I j I I, I l I I l I I I I I I I I I I I .1 ..1 I I I l I I I I I I I I I I I I I I I ~3 I I I I I I l I I I I I I I I I I I I I I I I I l I I I l I I I l I I I I I I I I I I I I I I 313 ! I I .I I I i I I I II I I I I I I I I I I I I j I I I i I I I j I I I I I I I I I I I I I I I 313 ' I I I I II j I I I I I I I I I I I I I I I I I i I I I j I I l__i_t I I I I I I I I I I I I I I 313 I II II j I I I ! I I I I I I I I I I I I I I I I --· j LLLI I I i I I I I I I I I I I I I I I I 313 I I ' I I I I I I ; I I I I I I I I I I I I I I I I I l I ..LLL I I i I . I I I I I I I I I I I I I I 313 I I J Co..on- Shop S1a1oon NO y.,,., P;,r3- STATION: CHLOROPHYLL ~ I meier ~" ~ "' ; • COUNTRY: 1 2 3 4 5 6 7 8 9"10 11 12 13 14 15 INSTITUTION· I I I I I I I 012 I~ TYPE MASTER RECORD I No orj Depth to bottom I Dolam- 2: volume S>Ze ~ Code N 1 1 E , , time (dnTl) Oi eter $ (cm3) (mm) ro 1 1 S' Q' W o Q' (GMT) ~ U I i1s 11h8 19 20 21 22 2312~!25 26l272B!293031i32 333413536137 38394041 424 444546 4748495051 5253154 55 56575859606162 636465665768 697011 n13 74 75 76 n[78[79BOj ~ I I I I I I I : I i I i I : I : I I i I i I L_LL_~j I I I I I I I I I .LL I I I I I I I I I I I I I I l I I~ DATA CYCLE RECORD~ ! - --~-· -- --·---- Un~s D No or Parall"l • 0 Chloro- I Qb-,ervatoon deplh -~ Unassogned Ph3-e0- Unenigned ~ w paral'el ~mp« ", z. phy11 _! pogment c- ··~ D No. . ;, 0 0 , (mg m-3) (mg..{.l) -- "''"'~ !,, u . z ~ ~ 16 17 REMARKS ~ 18 19 20 21 22123 24 25 26 27 2829 30 31 32 33 343536 37 38 3940 142 4344 4546147 4849150 st 525354555657 58596061626364656667686970 n n 737475 15n 78 7900

~ _l_j II l LLLLLLI._LLL.J.LL L _l_,_ ... _L! I l __ __ L_L_LLLl__LLl_l_L_L..l_l I I I I I I I I I I I I 313 _,_L_ ' 313 1-~.Ll~-- -- _LLLLLLLLJ_LLLJ_ _L,_ __Ll _ __t_l- .... L.L.. LL.I I I I Ll_l_LL.L I I I I I I I I I I . . I I I __L_I_l_Lj_l_L.LLLLJ .... L .... L _LI-. - _d LL ___ L_J. LLl_l_ _L_LLL.L.LLL_Ll I I I I I I I I I I 33 ,_.l I I j I I _I 1 I I I I I .L_L.J. .L _Lj__ .LJ I l _LLLLLLL I I I I I I L I I I I I I I L i I I I 313 ~ l ()1 ' ._, I I I I j I I I I I I I I I I I... LL ..1 I I I j I j I LLI I IIIL_LLLl.l II I I I I II I I I I 313 l I II I I i I I I I I I I I LJ_I I I I l I l LJ I I I I I I I I I I I I I I I I I I I I I I I I 313 ' I I I I j 33 I I I I lj_l I I I I I ~- -~~-.j--LL__l_j__j_L_L LLLl I I I I l I I I I I I I I I I I I l Ll I I I _L_j_l I I I I I :I _: L 1 33 .LJ '~Leu~''''' '' ''''' '' ''' I II II I __ l_l_l_l_LLLLLL L l l_ _l_l_ _LL l l J l l_i_L_LLLLLLLl I I I I I I I I I I I 313 -~-'-11- ' I I I I __ ....LLLL L l __ LLLJ.. I I 1.. LL _j_L.LLL.. Ll Ll LLLLLLLL.JI I I I I U I 313 _L~·- L l /I .Li. I I I I I ; __[ I - _j_J__ I illllllll_LJIIII.JIIIllilll 313 IIIIIIILLU_lJ _ .

I I I l l __ L _L __L_j_ 313 l I I I I I .L.LLLL__l.. LL ~- --· _LL _LLLLLLLLLI I I I I I L.LLI I I I I I I I ' I I I I I l I I I I I I I I I I I I I j I l I I I I I I I I I I I I I I I I I I I I I I I I l 313 ' I I I I I I l 'IIIIIIIIII.U I I I I l I j _l_ll I II I I I I I I I I lj_J__LLLll I I I I 313 I ' ' I I I I j _[_ __ LL ._L j_L_u_l_LLJ_LLLl_j_l__ j I LL I I I I I I 33 I I I I I I 1 I I I LL.L ..L_ . ~ ~-- j___ 1 I ...L

I I I I I J _l I I I I I .LLL.LLLI I ) . __ ,_ __ LL f-1-j T L LL I I I I Ll I 1 I LJ I I I I I I I I I I l 1 3~3 I I I II I IIIIIUIIIIII _L .L. - ·-· _L_j__ ..LLj __ LLl_l I I I I I I i I I I I I I I I I I I I I I I ~3

I .l I I I l I I ''I I I I I I I I I 11 L I I I I I I I 313 -160- 161

, __ ., ~~~-~ '".C'!I STATION · Z006ENTHOS 12. Microbiological data reporting format COVN"T~Y; l.I507301 .~7:L .. ' ' , I , , I lo.si INSTITUTION: ' "''YPE' I.V.SHR RECORD The microbiological data reporting format will be prepared by ICES, as requested by the Helsinki Commission (HELCOM 9/16, Paragraph 6.9). Ooi.T ' cYCLE RECDfiO ~-- ] "~ OF SAM"-ES• "•·~:--4":/1 '"' .. I BOTTOM TYPE: I::;:; -1•1 - .. ·~!i ...... ! 1-I•IJ! 1010:1021221:. ~.fOS!>Q !><16556'676860 "107tin n 1• 75101 '-' e.oe1~2 " " '"' : : I I ' I I I ; ' . ' I I I ' ' ' i I I ; I : I ' ' ' I !_c ._I_ ; ' ; : I ' ' ' I .. l i I ! ; ' : I I ; I I ' ' I ! I I : I ' ' , ! : : ' I I I ! ' I , I I : ' : : ' ' I 1 ; ; ' ' i , I Li; I ; I ' ' ' I __ , t I i j I j I I ' I ; ' ' I I ' ' I ' ! ; ' : ' ; : ' ' ' ' i ! ! . ! : I ' ' I I ' I I I 1 I I I I I I I . ' I ' ' ' ' ' ' ' ' I ' ' ' I : I I I I ; ' ; i 1 i : ' I I ; I I ' ' ' : ' ' ! ; I : ' I I I I ' I : ' ! ; ; I I ' : ' I ' I I I I ' I ' : ' ' ' ' ' ' I ' I I I ; I I . ' i I ' ' ' ' ' ' ' ' : 1 i .l I I __j_ ' I I , ; , I : ' ; I I ' , ' i ! ,_, i ! I .1 ' ' I ! ' ' I ' ' I ' , I ' I _, _l_ \ ; I : I I ' I ' I I ' ' I ' ' ' ' I 1_1 l J._j __j_ i ; I I ' _j ' L ' ~ ; ' ; ' ' j ' ' ' ' ' ' .: i I I ' I ! Li_ i ' ' l I I I ! ; I I I ' ' ' ' . ' ' ' i ' I I I l .. J l I ; ' ' ' ~ ' I ' i ' i ' i I I I : : ,i _! I J _I I J I i ; 1 i ' I ! ! ' ' ' ' ' I : ' : ' ' ' ' I I ! : I i ' ' I ' i ' ' ' ' ' I I I j ; I : ' I' I ' . : ' ' ' ' I i , I I ! I i i ' ' ' ' I I I I • I i ' I I i I I ' ' ' ' ' I ! I_!_ , . I i I ! I I ' II i I I : ; ' ' ' I ' ' ' . I I . 1_, 1 ..l ; ' . I : I ; I I : ! I ' ' ' ' I I ! [ ; I : i I ' ' ' J I I I I I I ' ' ; ' ! ' ' I I ' I u i i I 1 i I ; ' : ! i . ' . I I,_, I ' ' ' ' ' I ! I I I I I : I ; I : I I ' ' ' ' ' ' ' ' : ' i : : I " ! , I J .. LL i : , I : ' ' I ' ' ' ' ! ' I : I ; ! I i I i ; 1 I I I I~J ' l ' ' ' ' ' ' ' , __ L ; J • 1 ; I I I ' ' ' I I , i ' ' ' ' ' ' , " " I ' I I i i I : , ; ; ' : I j : I! u L1 ' ' ' ! ' ' ' ' ' I I t I ; ' ; I : I . ' I ' I I.~ ' i ' ' I i i I ; I ' ' I ' ' i I i ' I ; ' : I ' ' ' ' : ! : I I I I ' , ' I ' I I I I ll I ; ' i ' I ' ' ' ' : ' I I I ; , I I I I I I . ' ' I ' • i i ' ' ' ' ' BALTIC SEA ENVIRONMENT PROCEEDINGS No. 9 SECOND BIOLOGICAL INTERCALIBRATION WORKSHOP Marine Pollution Laboratory and Marine Division of the National Agency of Environmental Protection, Denmark, No. 1 JOINT ACTIVITIES OF THE BALTIC SEA STATES WITHIN THE August 17-20, 1982, R0nne, Denmark FRAMEWORK OF THE CONVENTION ON THE PROTECTION OF THE (1983) MARINE ENVIRONMENT OF THE BALTIC SEA AREA 1974-1978 (1979)* No. 10 TEN YEARS AFTER THE SIGNING OF THE HELSINKI CONVENTION National Statements by the Contracting Parties on the No. 2 REPORT OF THE INTERIM COMMISSION ( IC) TO THE BALTIC Achievements in Implementing the Goals of the MARINE ENVIRONMENT PROTECTION COMMISSION Convention on the Protection of the Marine Environment (1981) of the Baltic Sea Area (1984) No. 3 ACTIVITIES OF THE COMMISSION 1980 - Report on the activities of the Baltic Marine Envi­ No. 11 STUDIES ON SHIP CASUALTIES IN THE BALTIC SEA 1979-1981 ronment Protection Commission during 1980 Helsinki University of Technology, Ship Hydrodynamics - HELCOM Recommendations passed during 1980 Laboratory, Otaniemi, Finland (1981) P. Tuovinen, V. Kostilainen and A. Hlmlllinen (1984) No. 4 BALTIC MARINE ENVIRONMENT BIBLIOGRAPHY 1970-1979 (1981) No. 12 GUIDELINES FOR THE BALTIC MONITORING PROGRAMME FOR THE SECOND STAGE No. SA ASSESSMENT OF THE EFFECTS OF POLLUTION ON THE NATURAL (1984) RESOURCES OF THE BALTIC SEA, 1980 PART A-1: OVERALL CONCLUSIONS No. 13 ACTIVITIES OF THE COMMISSION 1983 (1981)* - Report of the activities of the Baltic Marine Envi­ ronment Protection Commission during 1983 including No. SB ASSESSMENT OF THE EFFECTS OF POLLUTION ON THE NATURAL the Fifth Meeting of the Commission held in Helsinki RESOURCES OF THE BALTIC SEA, 1980 13-16 March 1984 PART A-1: OVERALL CONCLUSIONS - HELCOM Recommendations passed during 1983 and 1984 PART A-2: SUMMARY OF RESULTS (1984) PART B: SCIENTIFIC MATERIAL (1981) No. 14 SEMINAR ON REVIEW OF PROGRESS MADE IN WATER PROTECTION MEASURES No. 6 WORKSHOP ON THE ANALYSIS OF HYDROCARBONS IN SEAWATER 17-21 October 1983, Espoo, Finland Institut fUr Meereskunde an der Universitlt Kiel, (1985) Department of Marine Chemistry, March 23 - April 3, 1981 No. 15 ACTIVITIES OF THE COMMISSION 1984 (1982) - Report on the activities of the Baltic Marine Envi­ ronment Protection Commission during 1984 including No. 7 ACTIVITIES OF THE COMMISSION 1981 the Sixth Meeting of the Commission held in Helsinki - Report of the activities of the Baltic Marine Envi­ 12-15 March 1985 ronment Protection Commission during 1981 including - HELCOM Recommendations passed during 1984 and 1985 the Third Meeting of the Commission held in Helsinki (1985) 16-19 February 1982 - HELCOM Recommendations passed during 1981 and 1982 No. 16 WATER BALANCE OF THE BALTIC SEA (1982) A Regional Cooperation Project of the Baltic Sea States; International Summary Report No. 8 ACTIVITIES OF THE COMMISSION 1982 (1986) - Report of the activities of the Baltic Marine Envi­ ronment Protection Commission during 1982 including No. 17A FIRST PERIODIC ASSESSMENT OF THE STATE OF THE MARINE the Fourth Meeting of the Commission held in ENVIRONMENT OF THE BALTIC SEA AREA, 1980-1985; GENERAL Helsinki 1-3 February 1983 CONCLUSIONS - HELCOM Recommendations passed during 1982 and 1983 (1986) (1983) No. 17B FIRST PERIODIC ASSESSMENT' OF THE STATE OF THE MARINE ENVIRONMENT OF THE BALTIC SEA AREA, 1980-1985; BACKGROUND DOCUMENT (1987) * out of print No. 18 ACTIVITIES OF THE COMMISSION 1985 - Report on the activities of the Baltic Marine Envi­ ronment Protection Commission during 1985 including the Seventh Meeting of the Commission held in Helsinki 11-14 February 1986 - HELCOM Recommendations passed during 1986 (1986)*

No. 19 BALTIC SEA MONITORING SYMPOSIUM Tallinn, USSR, 10-15 March 1986 (1986)

No. 20 FIRST BALTIC SEA POLLUTION LOAD COMPILATION (1987)*

No. 21 SEMINAR ON REGULATIONS CONTAINED IN ANNEX II OF MARPOL 73/78 AND REGULATION 5 OF ANNEX IV OF THE HELSINKI CONVENTION National Swedish Administration of Shipping and Navigation; 17-18 November 1986, Norrkoping, Sweden (1987)

No. 22 SEMINAR ON OIL POLLUTION QUESTIONS 19-20 November 1986, Norrkoping, Sweden (1987)

No. 23 ACTIVITIES OF THE COMMISSION 1986 - Report on the activities of the Baltic Marine Envi­ ronment Protection Commission during 1986 including the Eighth Meeting of the Commission held in Helsinki 24-27 February 1987 - HELCOM Recommendations passed during 1987 (1987)*

No. 24 PROGRESS REPORTS ON CADMIUM, MERCURY, COPPER AND ZINC (1987)

No. 25 SEMINAR ON WASTEWATER TREATMENT IN URBAN AREAS 7-9 September 1986, Visby, Sweden (1987)

No. 26 ACTIVITIES OF THE COMMISSION 1987 - Report on the activities of the Baltic Marine Envi­ ronment Protection Commission during 1987 including the Ninth Meeting of the Commission held in Helsinki 15-19 February 1988 - HELCOM Recommendations passed during 1988 (1988)

* out of print Baltic Marine Environment Protection Commission - Helsinki Commission - Mannerheimintie 12 A SF-00100 Helsinki

ISSN 0357-2994

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