POLISH JOURNAL OF ECOLOGY (Pol. J. Ecol.

56

379389

2008

Regular research paper

Maciej WALCZAK
Department of Environmental Microbiology and Biotechnology Institute of Ecology and Environment Protection, Nicolaus Copernicus University Gagarina 9, 87-100 Toru, Poland, e-mail: walczak@umk.pl

DAYTONIGHT ACTIVITY OF BACTERIA IN THE SURFACE MICROLAYER OF EUTROPHIC LAKE

ABSTRACT: This study examined changes of bacteria numbers in the surface microlayer (SM) and subsurface water (SW) of a lake during a day- and night-time. The research also addresses the synthesis of DNA and cell protein as well as the activity of cellular dehydrogenases depending on time of the day. Results demonstrated that in spring and summer the numbers of bacteria (per cm3) in the SM was significantly greater during night-time than day-time (average: May, daytime 30.058 106, night-time 71.343 106; July, day-time 10.801 106, night-time 40.353 106). In October, numbers of bacteria in dayand night-time were not statistically different (respectively: 5.841 106 and 3.664 106). Results indicated also that the rate of DNA synthesis by SM bacteria was much higher in the night-time (average: May 2.049 106 pg h1 cell1; July 1.363 106 pg h1 cell1), than in the day-time (average: May 0.7263 106 pg h1 cell1; July 0.3404 106 pg h1 cell1). In contrast, in October the values of DNA synthesis by SM bacteria were higher in night-time. These changes are significantly smaller in SW at a depth of several dozen centimetres. However, no significant impact was observed of a time of the day on the activity of protein synthesis and activity of cellular dehydrogenases by bacteria inhabiting SM and SW. KEY WORDS: surface microlayer, heterotrophic bacteria, day-night impact on bacteria

1. INTRODUCTION The surface microlayer (SM) covers of the earths surface and at the same time includes an infinitely small volume of the earths total water mass. This layer constitutes a specific chemical and physical environment, which differs substantially from the subsurface water (SW). As a result of such phenomena as adsorption, diffusion, flotation or rainfall, all kinds of organic matter, mostly lipids, proteins, polysaccharides and their derivatives accumulate in this layer, creating a superficial film or biofilm (Joi n and Mor is 1982, Kor z enie wsk i 1990, Pl as quel le c et al. 1991, Kost r z e wska-Szla kowska 2003). The film separates the hydrous habitat from the air. The presence of diverse chemical compounds in SM, generally occurring in higher concentrations than in SW (Fa l kowska 2001, Hi l lbr icht-Ilkowska and Kos t r z e ws k a - S z l a kows k a 2004) enhances the accumulation of both autotrophic (Kost rz e wska- Szl a kowska 2000) and heterotrophic organisms (Hopp e 1986) The surface microlayer generally contains an elevated number of bacteria, called bacterioneuston. Bacterioneuston both con-

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sumes and produces organic substances contributing in this way to the development of the surface microlayer. The physical stability of this layer is maintained by surface tension of the water surface. On the other hand, this environment is highly unstable in comparison to the subsurface water. This instability is caused by extreme temperature or solar energy variations. Due to the fact that bacterioneuston inhabits the surface microlayer, its members are exposed to stressful ecological conditions to a greater degree than organisms inhabiting the water column. Potential harmful factors, such as intense solar radiation, temperature, or the presence of toxic substances, play an important role in the struggle for survival and growth. All of these factors are selective and affect the composition of the microbiological assemblages (Shi lo 1979, Vest a l and Hobbie 1988). Among the physical and chemical factors determining the bioactivity of the surface water, the most important one seems to be the solar radiation. Light reaching the water surface may penetrate the water column down as far as several dozen meters. However, the highest light intensity occurs within the upper dozen centimeters. The amount of absorbed and diffused solar radiation varies and depends on the concentration and type of organic matter present in the water column. At high concentrations of dissolved organic matter, which contains a considerable amount of humic substances, harmful UVB radiation penetrates only the upper several centimeters of the water (Hess en et al. 1997). According to Z ait s e v (1971), the upper 10 cm of the water column absorbs ca. 75% of the UV radiation at = 254 nm. Considering the entire range of solar radiation reaching the air-water interphase, medium wave UV radiation, i.e. UVB 290320 nm and UVA 320400 nm, is of the highest biological importance due to its harmful effects. Radiation within this range causes DNA damage (lethal effect) or limits the growth of organisms by inhibiting enzyme synthesis, reducing active transport, or by inducing mutations. The 24-hour cycle of the solar radiation entails an entire sequence of changes taking place in the surface water. As a result of the natural changes of the intensity of the solar

radiation within twenty-four hours, the concentrations of many organic and inorganic compounds of the SM water as well as the numbers and the activity of both the phytoand the zooplankton (Fa l kowska 2001) vary adequately. This is why the solar insolation is also one of the main factors working indirectly and directly on the numbers and the activity of bakterioneuston. Despite numerous studies reporting potentially unfavorable impacts of light on bacterioneuston, many empirical studies exist which fail to demonstrate differences in neuston activity with and without solar exposure (Her manss on and D a h lb ck 1983) and also report an insignificant impact of UV and visible light on total bacterioneuston activity (G ar ab e t i an 1991, Wi l l i ams et al. 1986). On the other hand, there are numerous studies that demonstrate that solar radiation, especially UVB radiation, is detrimental to the production of bacterial biomass and exoenzyme activity (Herndl et al. 1993, B o av id a and We t z el 1998). It is also noteworthy that photooxidation of dissolved organic matter (DOM) and particulate organic matter (POM), which results in the release of considerable quantities of easily assimilable organic matter and may increase the activity of bacterioplankton (Her nd l et al. 1997), occurs under the influence of UV. This study examined the dynamics of numbers of bacteria in SM and SW as well as their metabolic activity in twenty-four hour cycle, depending on light conditions. 2. MATERIAL AND METHODS The studies were carried out in the eutrophic, small lake, (local name Jeziorak May), area 26 ha, maximal depth 6.4 m (Table 1). The lake has no inlets or outlets, but in its northern section is connected to another lake by a narrow and shallow (1.5 m) strait. The lake waters are considered as strongly eutrophic with hypertrophic symptoms due to water blooming and oxygen relations (Z b ek 2005) Water samples used for analyses were collected in month: May, July and October of 2005 from two stands localized in pelagic zone. Water of the surface microlayer was collected using plexiglas plate, which collects a

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Table 1. Morphometric and trophic characteristics of eutrophic lake (lake Jeziorak May) under study.
Characteristic Area (ha) Maximal depth (m) Mean depth (m) pH
(1)

Value 26 6.4 3.4 7.39.0 0.31 2.70 134380

3 (2)

Total phosphorus (mg dm3) (2) Total nitrogen (mg dm ) Electrolytic conductivity (S cm1) (1)

(1) data supplied by Department of Environmental Microbiology and Biotechnology, Nicolaus Copernicus University (mean for surface water, spring, summer and autumn 2005) (2) data supplied by Department of Hydrobiology, Nicolaus Copernicus University (mean for surface water, spring, summer and autumn 2005)

150 m water layer, and a Garrett mesh (G arrett 1965) with a hole diameter of 200 m, which collects a 200 m water layer. The SM water samples, collected in this way were examined separately, but the results were averaged and subjected to analysis as an average measurements for SM. The subsurface water (SW) was collected from a depth of 20 cm using a sterile, glass pipette and an automatic pump Pippet-boy (De Ville). Water was collected every 3 hours over a 24-hour period. Samples were poured into sterile, glass containers, from which 10 ml subsamples were obtained for analysis. The samples were analyzed for total numbers of bacteria (TNB), numbers of metabolically active bacteria TNAB (with an active electron transport system), activity of cellular dehydrogenases, and rates of protein and DNA synthesis. In all of the above analyses, the sample incubation was carried out for 2 hours in lake water with appropriate reagents (in situ). The intensity of visible light and UVB were measured simultaneously (Photometer PMA 2200, Solar Light Co) with collection of the samples used for microbiological analyses (Table 2). Total numbers of bacteria (TNB) was determined by a direct enumeration method on membrane filters (Millipore) with a pore diameter of 0.22 m. Samples were dyed with acridine orange (Z immer mann 1977), and visualized under an epifluorescent microscope (Carl Zeiss Jena).

Numbers of metabolically active bacteria (TNAB) was determined following the method of Z i mmer mann et al. (1978). Activity of cellular dehydrogenases (ACdH) was examined with the INT (2-p-iodophenyl-3-p-nitrophenyl-5-phenyl tetrazolium chloride) method, in which the amount of triphenyl formazan (TF) produced from the colorless substrate INT as a result of the activity of cellular dehydrogenases was measured. The color intensity is proportional to the activity of cellular dehydrogenases. Analysis of TF concentration was carried out on a spectrophotometer (Marcel Pro) in relation to a standard curve. Rate of cell protein synthesis (CPS) was analyzed by measuring the rate of incorporation of tritium labelled leucine, following Ki rchman , K Ness and Huds on (1985). The amount of generated protein was determined using a conversion factor based on an assumption that leucine constitutes on average 0.073% of bacterial protein mass (Ki rch man et al. 1989). Rate of DNA synthesis was determined by measuring the rate of incorporation of tritium labelled thymidyne (C hrst et al. 1998]). Statistical analyses were done using program STATISTICA 6.0. Analysis of Variance (ANOVA) was the primary statistical method used in calculations. This method facilitated comparison of the following independent factors: TNB, TNAB, activity of cellular dehydrogenases, rate of protein and DNA synthesis in day and night periods.

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Table 2. Daily changes of the intensity of solar light, UVB radiation and temperature in surface microlayer (SM) and subsurface water (SW) for three periods. Day-time: May and July 6.0018.00; October 9.0015.00; Night-time: May and July 21.003.00; October 18.006.00.
July Temp. (C) SM SW SM SW SM SW SM SW SM SW SM SW SM Light (klx) UVB (W cm2) Temp. (C) Light (klx) UVB (W cm2) Temp. (C) SW October

May

Sampling time SW

Light (klx)

UVB (W cm2)

SM

SW

SM

3.00

0.32

0.01

0.05

0.00

18.90

18.90

0.00

0.00

0.00

0.00

22.40

22.70

0.00

0.00

0.00

0.00

13.70

14.00

6.00

40.00

9.00

5.88

0.29

18.80

19.20

14.00

6.00

1.50

0.33

22.60

22.60

4.00

1.00

0.34

0.07

14.00

13.70

Maciej Walczak

9.00

82.00

46.00

11.92

1.48

18.60

18.50

30.00

22.00

3.37

1.22

22.60

22.50

14.00

5.00

1.19

0.38

14.30

13.80

12.00

125.00

60.00

18.11

1.93

18.60

18.70

40.00

28.00

6.00

1.55

23.00

22.70

20.00

8.00

1.71

0.61

14.20

13.90

15.00

76.00

6.00

11.00

0.19

19.70

19.70

38.00

26.00

4.56

1.32

23.50

22.90

22.00

10.00

1.88

0.76

14.30

14.10

18.00

10.00

8.00

1.42

0.25

19.80

20.20

15.00

8.00

1.68

0.44

23.20

23.20

0.10

0.00

0.00

0.00

14.20

14.30

21.00

0.04

0.00

0.00

0.00

19.40

20.00

0.80

0.10

0.09

0.00

23.10

23.10

0.00

0.00

0.00

0.00

14.00

14.30

24.00

0.00

0.00

0.00

0.00

19.00

19.20

0.00

0.00

0.00

0.00

22.80

23.00

0.00

0.00

0.00

0.00

13.70

14.20

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3. RESULTS Comparison of TNB values estimated in SM during the day-time and night-time demonstrates that in May and July, TNB is significantly greater in night (results of statistical analyses are presented in Table 3). In May, in the day-time, the average total numbers of bacteria (TNB) equaled 30.058 106, while at night-time, this number was much higher and equaled 71.343 106 cells cm3 (Fig. 1). An analogous relationship was observed in July; the average TNB in day-time equaled

10.801 106, while this number increased to 40.353 106 cells cm3 at nigh-time. In contrast, in October the mean values of the total numbers of bacteria at night-time and in daytime were not statistically different (5.841 106 and 3.664 106 cells cm3, respectively). However, this daily cycle was not observed in SW samples collected at the same time as SM samples (Fig. 1). The changes observed in TNAB in twenty-four hour cycle are largely analogous to changes in TNB (Fig. 2). In the SM water in May the average value of TNAB in day-

Table 3. Statistical differences (P-values) in investigated parameters in surface microlayer (SM) and subsurface water (SW) between day and night periods. See Fig. 15 for the values. Significant differences are given in bold.
Month May July October Total numbers of bacteria SM 0.019 0.036 0.052 SW 0.078 0.394 0.001 Total numbers of active bacteria SM 0.037 0.048 0.699 SW 0.359 0.743 0.949 Dehydrogenases activity SM 0.186 0.681 0.422 SW 0.490 0.808 0.071 Cell protein synthesis SM 0.543 0.383 0.347 SW 0.688 0.991 0.818 DNA synthesis SM 0.004 0.004 0.018 SW 0.017 0.218 0.013

Fig. 1. Total numbers of bacteria (TNB) in surface microlayers 150200 m (SM) and in subsurface water 20 cm (SW) in day and night periods. Vertical bars represent standard deviation.

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cells 10 6 cm-3

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cells 10 6 cm-3

Day

Night

Fig. 2. Total numbers of metabolically active bacteria (TNAB) in surface microlayer 150200 m (SM) and in subsurface water 20 cm (SW) in day and night periods. Vertical bars represent standard deviation.

mol TFh-1 cell-1

Day

Night

Fig. 3. Activity of cellular dehydrogenases (ACdH) in surface microlayers 150200 m (SM) and in subsurface water 20 cm (SW) in day and night periods. Vertical bars represent standard deviation.

time equaled 8.35 106 cells cm3, while at night-time this value increased to 17.14 106 cells cm3. Similarly, elevated numbers of metabolically active bacteria in the nighttime were also observed in July. During that

month, in the day-time, the average TNAB value equaled 7.17 106 cells cm3, while the average value at night-time was much higher and equaled 21.96 106 cells cm3. In contrast, such changes in the number of meta-

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ng 106 h-1 cell-1

Day

Night

Fig. 4. The rate of protein synthesis (CPS) by bacteria inhabiting surface microlayer 150200 m (SM) and subsurface water 20 cm (SW) in day and night periods. Vertical bars represent standard deviation.

bolically active bacteria related to twentyfour hour cycle were not observed during October. The mean TNAB values in the SM in October were similar and equaled 1.38 106 cells cm3 during day-time and 1.07 106 cells cm3 at night-time. In subsurface water, the number of metabolically active bacteria was not correlated with the time of the day (Fig. 2). The activity of cellular dehydrogenases (ACdH) in both SM and SW samples did not vary significantly in twenty-four hour cycle (Fig. 3). In May and October, the ACdH in the SM layer was higher at night-time than in day-time; however, these differences were not statistically significant. In contrast, during July the activity of the SM bacteria was higher in day-time than during night. In the SW, the activity of bacterial dehydrogenases (ACdH) did not vary in twenty-four hour cycle, and only in October the value in SM was significantly higher in day-time. In May and July, protein synthesis (CPS) for the SM bacteria was higher at nighttime (in May: day-time 0.8002, night-time 1.3224; in July: day-time 0.0758, night-

time 0.1944 106 ng protein h1 cell1). However, the differences were not considerable and this relationship was not statistically significant. In contrast, in October the CPS by bacteria from the surface microlayer was higher in the day-time, but the observed differences were small and statistically nonsignificant (day-time 1.1762; night-time 0.7194 106 ng protein h1 cell1). When analyzing the CPS in the subsurface water (Fig. 4) in subsequent seasons, no correlation was observed in twenty-four hour cycle. Average values obtained in a given month, in daytime and at nigh-time, were very similar. The rate of DNA synthesis by SM bacteria was much higher at night-time during May and July. In May, the average quantity of synthesized DNA per active bacterial cell in the SM in day-time and at night-time equaled 0.7263 and 2.049 106 pg h1 cell1, respectively. In July, this dependence also was clear and statistically significant. During this month the average amount of synthesized DNA per metabolically active bacterial cell equaled 0.3404 106 pg h1 cell1 in day-time. At night-time, the average value of

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pg 106 h-1 cell-1

Day

Night

Fig. 5. The rate of DNA synthesis by bacteria inhabiting surface microlayer 150200 m (SM) and subsurface water 20 cm (SW) in day and night periods. Vertical bars represent standard deviation.

DNA produced increased to 1.363 106 pg h1 cell1. In contrast, in October, the rate of DNA synthesis by SM bacteria was opposite, i.e. higher activity of DNA synthesis was observed in day-time. In the SW, where the quantity of incoming radiation is less, particularly in the case of UV, a reduced rate of DNA synthesis was noted only in May in day-time (day 0.6492; night 4.068 106 pg h1 cell1). In October, DNA synthesis by the SW bacteria was greater in day-time. In July, differences in the rate of DNA synthesis by SW bacteria in twenty-four hour cycle were so minimal that they were not statistically significant. 4. DISCUSSION The research demonstrated that day/night cycle affects both the total numbers of bacteria (TNB) and the numbers of bacteria with an active electron transport system (TNAB) in the SM water. The abundances of these bacteria decreased in the day-time. C hrst and Faust (1999) obtained similar results.

Despite the fact that these changes were clear and significant, their explanation is still unsatisfactory. The question could be raised as to whether reduced numbers of bacteria in the SM water during day-time is an effect of the migration of bacteria into deeper water layers or of lower reproductive activity or perhaps of more complex relationships connected with influence of light on primary production and bacterivores activity. Information provided in the literature is also inconsistent. Some researchers suggest that solar radiation has no significant impact on the numbers of bacteria in the SM, or that such impact is very limited (D a lb ck 1983, D enward et al. 1999, Skrc z e wsk i and Mud r y k 2003). The activity of DNA synthesis by bacteria cells was examined in this study in twentyfour hour cycle. Obtained results were supposed to make up a basis for interpretation of TNB and TNAB changes undergoing within twenty-four hours. If bacteria migrations or bacterivores activity caused observed fluctuation, the degree of DNA synthesis by SM bacteria should be roughly the same during the day.

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However, results presented in this study regarding the rate of DNA synthesis demonstrate clearly that this synthesis is limited in the SM water in day-time. Numerous researchers (Davidson 1998, Her nd l et al. 1997, Kais er and Her nd l 1997, C hrst and Faust 1999) have obtained similar results, reporting that DNA synthesis in bacterial cells was inhibited during day-time. According to C hrst and Faust (1999) and D av ids on (1999), solar radiation with UV hindered incorporation of 3H thymidyne from 15 to 50% in comparison to the control (night-time). In the present study, the reduction of DNA synthesis was observed to equal 65% in May and 75% in July in relation to the value obtained in SM at night-time. Comparison of DNA synthesis by bacteria in SM and SW in the day-time (July) reveals lower activity of DNA synthesis in SM of as much as 94%. Above differences of DNA synthesis might partly be explained by the fact that the SM bacteria numbers changes between dayand night-time. Perhaps observed decrease of DNA synthesis by SM bacteria is not only a result of UV unfavourable impact, but this radiation is either the most important or one of the most crucial factors that hamper DNA synthesis. The greater reduction of DNA synthesis, in comparison to investigations of Kais er and Her nd l (1997) as well as of C hrst and Faust (1999), observed here results from the fact that the thickness of the sampled SM water equaled 100300 m. Whereas, the cited studies sampled surface water over a thickness of several dozen centimeters. The difference in sampling depth is of fundamental importance, due to the quantity of radiation, and in particular the UV radiation, penetrating the water. It is notable that in subsurface water samples, collected from a depth of ca. 20 cm, a significant impact of solar radiation on the rate of DNA synthesis was not observed. This is a result of the fact that the analyzed water originated from a eutrophic lake characterized by high concentrations of dissolved and particulate organic matter (DOM and POM), which absorbs UV radiation intensively (C o ckel l 2000). Therefore, the quantity of UV radiation reaching the SW was significantly reduced in comparison to SM (Table 1). The studies cited above were

conducted in marine ecosystems where the concentration of organic matter was much lower and the penetration depth of UV radiation greater. A significant impact of the day-time or night-time on the rate of bacterial cell protein synthesis (CPS) was not observed in the present study. However, it should be noted that the rate of CPS by the SM bacteria was greater at night-time, though these differences were not large and were not statistically significant. Results presented by Her nd l et al. (1997), Kais er and Herd l (1997) as well as C hrst and Faust (1999), demonstrate that absorption of 3H leucine by bacteria was also inhibited during day-time. However, it should be noted that the investigations of other authors were carried out at different latitudes. It pertains in particular to C hrst and Faust (1999) who conducted their research in the subequatorial zone where the solar radiation intensity is much greater than in temperate zone. Furthermore, the results presented by Kais er and Her nd l (1997) as well as C hrst and Faust (1999) indicated that the effect of solar radiation on CPS is less significant than on DNA synthesis. Based on these results, it can be inferred that the synthesis of DNA containing thymine, which easily creates dimmers under the influence of UV, is more sensitive to radiation than the synthesis of protein. Protein synthesis occurs on the RNA matrix, where uracil is present instead of thymine, and this feature potentially reduces the negative effect of UV on protein synthesis. This study also includes the results on changes in the activity of cellular dehydrogenases (ACdH) in twenty-four hour cycle. The present analysis demonstrated that the time of the day has no significant impact on activity of cellular dehydrogenases of bacteria from either the SM or SW. Unfortunately, no prior results of studies examining the changes of activity of these enzymes in twenty-four hour periods have been found in the available literature. However, according to research conducted by B o av id a and We t z el (1998), extracellular phosphatase activity was highly inhibited under the influence of solar radiation, during day-time. The activity of -glucosidase and urease also was inhibited by several dozen percent

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in the presence of strong solar radiation (Jorgens en et al. 1998). However phosphatase and glucosidase are categorized as exoenzymes, and are secreted to the external environment, where there is no protection against the harmful effect of UV. In contrast, dehydrogenases are intracellular enzymes, and are protected by cellular shields or caroteinoids inside the cell. Furthermore, it is difficult to unambiguously confirm that UV did not limit the activity of these enzymes. Measured ACdH can result from two types of UV effects. As in the case of extracellular enzymes or DNA, UV may inhibit dehydrogenase activity. On the other hand, photooxidation of organic matter in water occurs under the influence of UV radiation (Hder et al. 1998). This effect considerably increases the quantity of organic matter easily accessible to bacteria, and this must cause an increase in the activity of dehydrogenases participating in oxidation of organic compounds. Thus, it seems that the observed lack of differences in enzyme activity in day/night cycle constitutes a result of many physical and chemical factors, including UV radiation. The results testifying the indirect effect of insolation on the SM bacteria were presented by Fa l kowska (2001). In this work, a sequence of results was given, testifying the changes of concentrations of several kinds of nutrient compounds in the SM water and the changes concerning phytoplankton within twenty-four hours. The conducted research confirmed and expanded earlier reports regarding the important impact of day-time on overall activities of bacteria in aquatic environments. However, it should be noted that the effect of solar radiation, including UV, is not limited to simple and direct impacts on bacterial cells. Radiation also influences the environment through a wide range of indirect effects, e.g. organic matter photooxidation or the impact on phyto- and zooplankton and bacteriophages, which in turn affect the activity of aquatic bacteria. ACKNOWLEDGEMENTS: This work was supported by the Ministry of Science and Information Society Technologies under grant no. 0541/P04/2005/28

5. REFERENCES
B o av i d a M . J. , Wet z e l R . G . 1998 Inhibition of phosphatase activity by dissolved humic substances and hydrolytic reactivation by natural ultraviolet light Freshwater Biology, 40: 285293. Chrst R . J., Faust M.A. 1999 Consequences of solar radiation on bacterial secondary production and growth rates in subtropical coastal water (Atlantic Coral Reef off Belize, Central America) Aquatic Microbiol. Ecol. 20: 3948. Chrst R .J., O verback J., Wc iso R . 1998 Evaluation of the [3H]Thymidine method for estimatingbacterial growth rates and production in lake water: Re-examination and methodological comments Acta Microbiol. Pol. 1: 95112. C o cke l l C . S . 2000 Ultraviolet radiation and the photobiology of earths early oceans Origins of Life and Evol. of the Biosph. 30: 467499. D a h lb ck B. 1983 Microbial accumulation of interfaces. Marine micro organisms and interfaces Univ. of Gteborg. 3: 1215. D av ids on A.T. 1998 The impact of UVB radiation on marine plankton Mutation Res. 422: 119129. D enwar t C . M . T. , E d l i ng H . Tr anv i k L . J. 1999 Effects of solar radiation on bacterial and fungal density on aquatic plant detritus Freshwater Biology, 41: 575582. Fa l kowska L. 2001 12-hour cycle of matter transformation in the sea surface microlayer in the offshore waters of Gdask Basin (Baltic Sea) during spring Oceanologia, 43:201 222. G arab et i an F. 1991 14C-glucose ptake and 14 C-CO2 production in surface microlayer and surface-water samples: influence of UV and visible radiation Mar. Ecol. Prog. Ser. 77: 2126. G ar rett W. P. 1965 Collection of slick-forming materials from the sea surface Limnol. Oceanogr. 10: 602605. Hder D.P., Kumar H.D., Smit h R .C., Wor rest R .C. 1998 Effects on aquatic ecosystems J. Photochem. Photobiol. 46: 5368. Hess en D.O., Gj essing E.T., Knulst J., Fj eld E. 1997 TOC fluctuations in humic lake as related to catchment acidification, season and climate Biogeochemistry 36: 139151. Her manss on M., D ahlbck B. 1983 Bacterial activity at the air/water interface Microbiol. Ecol. 9: 317328.

journal 15.indb 388

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Day-to-night activity of bacteria in the surface microlayer of eutrophic lake Her ndl G.J., Br ug ger A., Hager S., Kais er E., Ob er nosterer I., Reit ner B. Slezak D. 1997 Role of utraviolet-B radiation on bacterioplankton and availability of dissolved organic matter Vegetatio, 128: 4351. Her nd l G . J. , Mu l l e r- Ni k l a s G . , Fr i ck J. 1993 Major role of ultraviolet-B in controlling bacterioplankton growth in the surface layer of the ocean Nature, 361: 717719. Hillbr icht-Ilkowska A., Kostrze wskaSzl a kowska I . 2004 Surface microlayer in lakes of different trophic status: nutrients concentration and accumulation Pol. J. Ecol. 52: 461478. Hopp e H. 1986 Degradation in sea water Biotechnol. 8: 453474. Joint R ., Mor r is R . 1982 The role of bacteria in the turnover of organic matter in the sea Oceanogr. Mar. Biol. Ann. Rev. 20: 65118. Jorgens e n N . O. G . , Tranv i k L . , E d l i ng H., Graneli W. , L idel l M. 1998 Effects of sunlight on occurrence and bacterial turnover of specific carbon and nitrogen compounds in lake water FEMS Microbiology Ecol. 25: 217227. Kais er E., Her ndl G.J. 1997 Rapid recovery of marine bacterioplankton activity after inhibition by UV radiation in coastal waters Appl. Environ. Microbiol. 10: 40264031. Kirchman D.L., Knees E., Ho ds on R .E. 1985 Leucine incorporation and its potential as a mesure of protein synthesis by bacteria in natural aquatic systems Appl. Environ. Microbiol. 3: 599607. Ki rchman D.L., Ne well S.Y., Ho ds on R .E. 1989 Incorporation versus biosynthesis of leucine: implications for measuring rates of protein synthesis and biomass production by bacteria in marine systems Mar. Ecol. Prog. Ser. 32: 4757. Korz enie wsk i K . 1995 Background of chemical oceanography Wydawnictwo Uniwersytetu Gdaskiego. (In Polish). Kos t r z e ws k a - S z l a kows k a I . 2000 Air-water interface in lakes: relations to in-lake trophism Verh. Internat. Verein. Limnol. 27: 18711874.

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Kostrze wska-Szla kowska I. 2003 A surface microlayer of waters: characteristics and research methods Wiad. ekol. 49: 183202 (In Polish). Plas quellec A., B eucher H., L e L ay C., C leret J. 1991 Quantitative and qualitative bacteriology of the marine water surface microlayer in a sewage polluted area Mar. Environ. Res. 31: 227239. Shi lo M. [E d.] 1979 Introduction. Strategies of Microbial Life in Extreme Environments Shilo M. Verlag Chemie, Berlin, pp. 1113. Skrcz e wsk i P., Mudr y k Z. 2003 Dynamics of daily changes to the number and production of estuarine bacterioneuston and bacterioplankton AUMC, Limnological Papers, 13: 5563. Vest al J. R . , Hobbie J.E. 1988 Microbial adaptations to extreme environmets. (in:) Microorganisms in Action:Concepts and Applications in Microbial Ecology Blackwell Scientific Publications, Oxford, 193206. Wi l l i ams P. M . , C ar lu c c i A . F. , Hen r i chs S.M., Van Vleet E.S., Hor r i gan S.G., Reid F.M.H., Rob er ts on K.J. 1986 Chemical and microbiological studies of sea-surface films in the southern Gulf of California and off the west coast of Baja California Mar.Chem. 19: 1798. Z aits e v Y.P. 1971 Marine Neustonology Israel Prog. Scientific Tranl. Jerusalem Z b ek E. 2005 Qualitative and quantitative changes of Blue-Green Algae in response to physicochemical water parameters in the urban lake Jeziorak May Oceanol. Hydrobiol. Studies, 39: 3956. Z immer mann R . 1977 Estimation of bacterial number and biomass by epifluorescence microscopy and scanning electron microscopy (In: Microbiol. Ecol. of Brackish Water Environ. Ed. G. Rheinheimer) SpringerVerlag, New York. pp. 103. Z immer mann R ., Itur r i aga R ., B e ckerBirck J. 1978 Simultaneous determination of the total number of aquatic bacteria and the number thereof involved in respiration Appl. Environ. Microbiol. 12: 926935.

Received after revising December 2007

journal 15.indb 389

2008-09-23 10:54:34