Romanian Biotechnological Letters Vol. 20, No.

3, 2015
Copyright © 2015 University of Bucharest Printed in Romania. All rights reserved
ORIGINAL PAPER

Benthic microbial biomass dynamics in aquatic

ecosystems of the Danube Delta

Received for publication, March 03, 2014

Accepted, May 16, 2015

IOAN PĂCEŞILĂ
Institute of Biology, Romanian Academy
*
Corresponding author: ioan.pacesila@gmail.com

Abstract
Sediments represent distinct ecological areas with a significant importance in transfer of matter
and energy through aquatic ecosystems. Many heterogeneous groups of heterotrophic microorganisms
coexist at sediment level and breakdown the detrital organic matter (DTOM) originated from excretes
and dead bodies of aquatic organisms. Through the metabolic activity of heterotrophic
microorganisms, the mineral elements from the DTOM composition are recycled and become available
to primary consumers, being reintroduced into biogeochemical cycles. Assessment of total microbial
biomass (TMB) can provide general information about the active microorganisms implied in energy
and matter transformation. This paper presents several characteristics of TMB dynamics, evaluated
during two research projects in sediments of several aquatic ecosystems from Danube Delta: three
sections of Sf. Gheorghe branch in 2008-2010 period, and Roşu Lake, Mândra Lake and two adjacent
channels in 2011-2013 period. The results show significant temporal and spatial variations of this
parameter, with different tendencies in the studied ecosystems. Highest TMB was recorded in Mândra
Lake (general average values of 966 g C g-1). Two statistical methods, hierarchical cluster analysis
and ANOVA, were used to determine the grouping by similarity of ecosystems and areas, and, also, to
evaluate the differences between obtained data series.

Keywords: total microbial biomass, detrital organic matter, microbial decomposition, Danube Delta

1. Introduction
The decomposition of detrital organic matter (DTOM) represents a key process in the
functioning of aquatic and terrestrial ecosystems. Through this process the mineral elements
present in the DTOM composition are recycled and, thus, become available to primary
consumers, being reintroduced into biogeochemical cycles. In this way is avoided the
depletion of mineral resources and excessive accumulation of organic matter (BOTNARIUC
& VĂDINEANU [2]; ZARNEA [29]; CERNAT & LAZĂR [3]), which would result in a
significant changing of the current forms of organization of living matter.
DTOM is decomposed especially by heterotrophic microorganisms, most of them
bacteria and fungus. Due DTOM degradation these microorganisms obtains energy and
mineral elements necessary for their metabolic activity. Although other categories of
organisms are involved in DTOM decomposition, the role of microorganisms is extremely
important because of their high rate of metabolic activity, rapid growth and multiplication,
large physiological diversity and ubiquitous presence in the environments (POMEROY et al.
[19]; NICOLESCU et al. [16];CERNAT & LAZĂR [3]).

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 Benthic microbial biomass dynamics in aquatic
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In aquatic ecosystems DTOM decomposition starts in the water column and continues in
sediments due to accumulation and sedimentation of organic particles (ZARNEA [29];
WETZEL [28]). The aquatic sediments represent distinct ecological areas (IONICĂ [10],
[11]) where many heterogeneous communities of heterotrophic microorganisms coexist
(ZARNEA [29]; WETZEL [28]). The sediment medium is the result of very different factors
like mineral composition, particle size, quantity and quality of organic molecules and
geographical localization (CURTICĂPEAN & DRĂGAN-BULARDA [4]) Its essential
contribution in ecosystem cycling, by physical, chemical and biological processes, it is
generally recognized. (REKHA & al. [22]). The sediment is also the main place where
degradation processes of refractory organic macromolecules, including pollutants, occurs
(CURTICĂPEAN & DRĂGAN-BULARDA [4]).
The most active part of sediment is the upper layer, at the water-sediment interface. At
this level organic molecules are breakdown through aerobic processes while in the deeper
layers of sediment the decomposition is preponderantly anaerobic, especially through
fermentation processes (ZARNEA [29]; WETZEL [28]). As benthic microbial communities
quickly respond to environmental perturbations, nowadays, many researchers consider them
as an early indicator of aquatic ecosystem stress (NWEKE et al. [18]).
Biomass of living microorganisms is an important component of total organic matter
present in an ecosystem (IGLESIAS [9]). Assessment of this structural parameter, with a
particular biological importance, provide us information about the amount of energy stored by
microbial communities existing in an ecosystem at a given time and, also, offer us a "picture"
of the active microorganisms implied in energy and matter transformation (VESTAL &
WHITE [27]; SIMON-GRUIŢĂ [24]).
In an aquatic ecosystem the dynamic of TMB is influenced by a number of factors like
physico-chemical parameters dynamics, the quality and the quantity of TDOM, predators and,
also, by competition for nutrients with algae and macrophytes (FINDLAY & SINSABAUGH,
[6]; BARTON & NORTHUP [1]; WETZEL [28]).
The Danube Delta is a dynamic heterogeneous macro-regional complex of aquatic, wetland
and terrestrial ecosystems. It is unique on the European continent, with a high biodiversity, and
can be considered an interface between the hydrographic network of Danube River and the Black
Sea. Throughout, along with Black Sea, Danube Delta is the main natural system with a very
important biogeochemical and hydrological role at the local and regional level (POSTOLACHE
[21]), and, also, have an obvious impact on economical and social development of local and
regional communities. During the last decades, like many other large wetland systems, Danube
Delta was under increasing anthropogenic pressure. The economical programs implemented by
authorities, whose purpose was the development of agriculture, industry and navigation have been
applied without taking into account the main functions of natural systems and, therefore, had a
negative cumulative effect over the environment, leading to hypertrophy of aquatic ecosystems
(FLORESCU & al. [7]; JUGARU & al. [13]).
In the present paper are shown some characteristics of spatial and temporal dynamics of
total microbial biomass (TMB) from sediments of several different aquatic ecosystems of the
Danube Delta.

2. Materials and Methods

The study was conducted in two distinct areas: Sfântu (Sf.) Gheorghe branch, during the
2008-2010 period, and two shallow lakes with different characteristics, Roşu Lake and
Mândra Lake, and their adjacent channels, during the 2011-2013 period.

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Sf. Gheorghe is the southernmost of the three main branches through which the Danube
flows into the Black Sea and carry about 21% of the total flow of the main river. In order to
improve the navigation, in the 1985-1990 interval this branch was shortened from 108 km to 70
km by cutting seven meanders along its length (FLORESCU et al. [7]). The hydrogeo-
morphological changes caused by this anthropogenic intervention led to apparition of three
distinct sections in the river branch: the free-flowing sector (FS), the meanders section (MS)
and the newly built channel (NBC). Samples were collected in April, July and October, from
seven sampling stations (Fig. 1a) located in the three mentioned sections, as it follows:
- the free-flowing sector (FS) - three stations: S1, S4 and S7;
- the meanders section (MS) - two stations: S2 and S5;
- the newly built channel (NBC) - two stations: S3 and S6.

Figure 1. Location of sampling stations: a) Roşu Lake (R), Mândra Lake (M),
Roşu - Împuţita Channel (C1) and Roşu – Puiu Channel (C2) and b) Sfântu Gheorghe branch

Roşu and Mândra are two representative shallowlake from the Danube Delta complex.
Lake Roşu is the youngest and the largest lake in the fluvial-maritime delta, with algal
blooming which can be considered signs of eutrophication (MOLDOVEANU & FLORESCU
[15]), while Lake Mândra is a low depth meso-europhic aquatic ecosystem (PARPALA &
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ZINEVICI [19]). Samples were collected in May, July and September from five stations in
Roşu (R1 – R5) and one station in Mândra (M). For a more accurate assessment, another two
sampling stations were located in two adjacent channels: Roşu – Împuţita Channel (C1) and
Roşu – Puiu Channel (C2) (Fig. 1b).
All samples were collected from the upper layer of the sediment, at the water-sediment
interface, with a Corer device (IONICĂ [11]). TMB concentration was determined by the
dosage of lipid phosphate present in the cellular membrane of microorganisms (IONICĂ et al.
[12]). The results are expressed in μg C per gram of dry matter.
The Paleontological Statistics (PAST) software (HAMMER et al. [8]) was used for
statistical analyses. All variables were ln (x+1) transformed.

3. Results and Discussions

The results show significant temporal and spatial variations of TMB, with different
tendencies in the studied ecosystems.
In Sf. Gheorghe branch, seasonal dynamics reveals the highest TMB values in the summer
for all sections (Fig. 2a), and the lowest TMB values in the spring for FS and NBC, and in the
autumn for MS. The increasing of temperature stimulates the microbial metabolism in the
superficial layer of the sediment and, in the same time, lead to the decreasing the oxygen
concentration at this level; a low oxygen concentration negatively affects the distribution of
benthic protozoa and invertebrates, the main consumers of microorganisms in sediment
(SUTTON & FINDLAY [25]). The highest annual average values of TMB were recorded in 2010
in all sections, while the lowest were recorded in 2009 for FS and MS and in 2008 for NBC.
Spatially, MS it seems was the most suitable environment for development of benthic
microbial communities (TMB general average value of 149 μg C g-1) in Sf. Gheorghe Branch.
Instead, in sediments of NBC the presence of heterotrophic microorganisms were much lower
(TMB general average value of 114 μg C g-1) than the other two sections (Fig. 2a). This
situation could be due to hydrogeomorphological changes caused by the anthropogenic
intervention, which, ultimately, affect the accumulation and erosion processes along this
branch (JUGARU et al. [13]). At present, in different regions of the cut off meanders there are
accumulation processes of silt, TMOD respectively, while the newly created channels are
characterized by intense erosion. These issues influence certainly silt transport system along
the river arm and also its geomorphology. Over time, the accumulation processes will lead to
decreasing the depth in the natural meanders, and the erosion processes will lead the
deepening of the newly created channels.
In the lentic ecosystems, Roşu and Mândra shallowlakes and, also, the adjacent Roşu –
Împuţita and Roşu – Puiu Channels, TMB spatial and temporal dynamics was characterized
by different tendencies. Seasonal average values were maximum in summer for Roşu Lake
and Roşu – Împuţita Channel, and in spring for Mândra Lake and Roşu – Puiu Channel. The
annual dynamics of TMB show the highest values in 2013 for Roşu Lake, in 2011 for Mândra
Lake and in 2012 for Roşu – Împuţita and Roşu – Puiu Channels (Fig. 2b). The highest
biomass of benthic heterotrophic microorganisms was recorded in Mândra Lake (TMB
general average value of 966 μg C g-1). This situation can be related to presence of aquatic
macrophytes which were relatively abundant in this type of aquatic ecosystems (KUHAR &
al. [14]; NURMINEN [17]; TOCCO & al. [26]). The DTOM result from the death of aquatic
macrophytes, especially in autumn, represent, after sedimentation, an important source of
carbon and energy for numerous categories of benthic heterotrophic microorganisms. In
contrast, in Roşu Lake submerged vegetation appears only sporadically (SÂRBU [23]).

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a)

b)

Figure 2. Seasonal (A) and annual (B) dynamics of TMB in: a) Roşu Lake (R), Mândra Lake (M),
Roşu - Împuţita Channel (C1) and Roşu – Puiu Channel (C2) and b) Sfântu Gheorghe branch’s sections,
the free flowing sector (FS), the meander section (MS) and the newly built channel (NBC)

Figure 3. The similarity (Ward's method) of the studied areas based on TMB dynamics;
Roşu Lake (R), Mândra Lake (M), Roşu - Împuţita Channel (C1) and Roşu – Puiu Channel (C2) and Sfântu
Gheorghe branch’s sections, the free flowing sector (FS), the meander section (MS) and the newly built channel
(NBC)

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Roşu – Puiu Channel was the ecosystem with less development of benthic microbial
communities (TMB general average value of 524 μg C g-1).
In order to evaluate the similarity between research areas about TMB variation in
studying periods, hierarchical cluster analysis, Ward’s method, was applied (Figure 3). From
the obtained dendrogram it can be observed that Sf. Gheorghe branch’s sections were grouped
separately from the other research areas. The different hydrogeomorphologic characteristics
of ecosystems and the particularity of physical and chemical parameters dynamics in studying
period can be an explanation for this arrangement. High similarity existed between MS and
FS areas of Sf. Gheorghe branch, and, also, between Roşu Lake and Roşu-Puiu Channel. The
dynamic of TBM in sediments of the Roşu – Împuţita Channel had a low similarity with the
dynamics of TMB from sediments of the other ecosystems studied in the same period.
The differences between TMB dynamics of research areas were evaluated using one way
ANOVA test. Data series were represented by all seasonal average values of each evaluated
ecosystem. The result (n=9, F=5,744, p<0,0001) confirmed the existence of significant
differences among the variables. Post – hoc test TUKEY is an adequate tool to determine
where these differences lie (DYTHAM [5]). The significant differences were obtained
between Mândra Lake and the Sf. Gheorghe branch’s sections, FS (Q=5,497, p<0,01), MS
(Q=5,674, p<0,01) and NBC (Q=6,15, p<0,01), and , also, between Roşu Lake and Sf.
Gheorghe branch’s sections, FS (Q=4,171, p<0,01), MS (Q=4,348, p<0,05) and NBC
(Q=4,824, p<0,05)

Conclusions
According to the obtained results it can be concluded that the TMB recorded high spatial
and temporal variations in the studied ecosystems.
In Sf. Gheorghe branch, the seasonal dynamic of TMB shows the highest values in
summer for all sectors. Also, the maximum developments of benthic microbial communities
were recorded in 2010. Spatially, TMB values were highest in MS, where accumulation of silt
occurs, and lowest in NBC, a sector characterized by the presence of intense erosion
processes at sediment level.
The lentic and lotic ecosystems studied in 2011-2013 period shows different patterns of
spatial and temporal dynamics. The summer season was the most favorable for the
development of heterotrophic microorganisms in the sediments of Roşu Lake an Roşu –
Împuţita Channel, while in the sediments of Mândra shallow lake and Roşu – Puiu Channel
heterotrophic microorganisms show maximum development in spring season. The highest
annual average values were recorded in 2013 for Roşu Lake, in 2011 for Mândra shallow lake
and in 2012 for Roşu – Împuţita and Roşu – Puiu Channels. Also, in the studying period, the
spatial dynamics of TMB show the highest values in Mândra shallow lake and the lowest
values in Roşu - Puiu Channel.
After applying of hierarchical cluster analysis, the obtained dendrogram revealed a
different grouping between Sf. Gheorghe branch’s sectors and Roşu and Mândra shallow
lakes and adjacent channels. The highest similarity was observed between FS and MS, and,
also, between Roşu – Lake and Roşu – Puiu Channel. The dynamic of TBM in sediments of
the Roşu – Împuţita Channel had a low similarity with the dynamics of TMB from sediments
of the others ecosystems studied in the same period.
The result of the ANOVA test (n=9, F=5,744, p<0,0001) indicates the existence of
differences between TMB seasonal dynamics evaluated from different Danube Delta
ecosystems in 2008 - 2013 period.

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Acknowledgment
This study was funded by the project no. RO1567-IBB02/2014 from the Institute of
Biology of Romanian Academy, Bucharest.

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