Aquatic Microbiology H.A.Foster revised November 2013.

Aquatic Microbiology

H.A.Foster

revised November 2013

2

Water

• 71% of the Earth’s surface is covered with water.

• 97% of this is marine and much of the freshwater is frozen.

• ¾ of the oceanic water is at a depth of >1000m

3

Physiological conditions

• Temperature below 300m relatively constant at approximately 3ºC. (remember ocean currents e.g Gulf stream)

• Pressure increases with depth (1 atm per 10m)

• Deepest parts are 11,000m (1100atm).

• Organism can be barotolerant or barophilic.

4

5

6

LENTIC

(Lakes, ponds, reservoirs)

LOTIC

Rivers and streams

Freshwater habitats

7

Biological Zones in a Lake

8

Microbiological habitats

• Planktonic - the water column, floating free in the water.

• Neustonic – the surface film.

• Benthic – in and on permanently submerged sediments.

• Epibiotic – On living surfaces.

• Sestonic – On floating organic matter e.g.faeces.

Food web in pelagic

zone

9

10

Colonisable surfaces within water

Epiphyton Surface of plants

Episammon Surface of sand or silt

Hyposammon Interstitial water

Epipelon Surface of sediment (mud)

Endopelon Inside the sediment

Epilithon Surface of stones

Endolithon Cavities in rocks/stones

Epizoon Surface of animals

Fouling Surface of artificial objects

11

12

13

• TemperatureTemperature is an important variable in

aquatic ecosystems:- changes the reaction rates (kinetics) of chemical and biochemical reactions - influences species distributions- controls dissolved O2 and CO2

- changes density of water

Greatest density of water occurs at 4°C below this temperature, water adopts a crystalline structure on its way to form ice. Ice has a much more open structure.

14

The changes in density with temperature cause stratification in lakes and physical turnovers.

Stratified lakes separate into 3 zones known as the:

EPILIMNION

HYPOLIMNION

THERMOCLINE (METALIMNION, or CHEMOCLINE)

15

Lake Stratification in Summer and Winter

Leon

ard

W. C

asso

n, P

h.D

., P.

E., D

EELe

onar

d W

. Cas

son,

Ph.

D.,

P.E.

, DEE

16

Holomictic lakes

• Show two cycles of mixing in spring and autumn (fall).– In winter a thermocline develops with water at

<4°C on top of water at 4°C.– In spring, the surface layer warms and the

thermocline disappears. This leads to mixing of the upper and lower layers and can lead to eutrophication and an algal bloom.

17

Holomictic lakes

– The upper layers then warm and during the summer the thermocline is re-established with warm surface water on top of colder waters.

– During autumn, the combination of cooling surface waters and mixing due to autumnal gales removes the thermocline and mixing and algal blooms can occur again.

– The thermocline leads to spatial distribution of organisms.

18

19

20

a) Holomictic (dimictic), two periods of circulation as described.

b) Amictic - sealed permanently by ice.

c) Cold monomictic - lake temp < 4°C (one period of circulation in summer).

d) Warm monomictic - lake temp > 4°C - circulates in winter; stratifies in summer; sub tropical regions, e.g. Florida.

Lake types

21

Lake types

e) Oligomictic - rare circulation (tropics).

f) Polymictic - frequent circulation.

g) Meromictic - does not undergo complete circulation due to stratification by

something other than temperature, e.g. salinity; can be caused by humans connecting sea and freshwater systems.

22

23

24

Food web for fast flowing waters

25

Bdellovibrio bacteriovorans

26

Hyphomicrobium sp.

27

Lakes and Rivers

• Water Pollution has different effects on lakes and rivers.

• Pollution of lakes and rivers can cause eutrophication.

• Because lake water is not quickly replaced the effects can accumulate gradually, in rivers pollution is eventually washed away to the sea.

• Waste, especially wastewater, from human or animal origin can contain pathogens.

28

Eutrophication is a natural process that occurs to all lakes over time as the weathering of rocks and soils from the surrounding catchment area leads to an accumulation of nutrients in the water and associated sediments.Young lakes (and man made reservoirs) usually have low levels of nutrients and correspondingly low levels of biological activity. Such lakes are referred to a being oligotropic from the Greek work oligos meaning little or few. Literally oligotrophic means little-nourished.Old lakes usually have high levels of nutrients and correspondingly high levels of biological activity. Such lakes are referred to as being eutrophic from the Greek word eu meaning well. Literally eutrophic means well-nourished.

Eutrophication

29

The main causes of eutrophication are:• natural run-off of nutrients from the soil and the weathering

of rocks.• run-off of inorganic fertiliser (containing nitrates and

phosphates).• run-off of manure from farms (containing nitrates,

phosphates and ammonia).• run-off from erosion (following mining, construction work

or poor land use).• discharge of detergents (containing phosphates).• discharge of partially treated or untreated sewage

(containing nitrates and phosphates).• In most freshwater lakes the limiting nutrient is phosphorus,

so an input of phosphorus in the form of phosphate ions (PO4

3-) results in an increase in biological activity.

30

The natural time scale for the aging of a lake from being oligotrophic to eutrophic is of the order of thousands of years. However, a high rate of input of nutrients (from human activities) can increase the rate of aging significantly resulting in eutrophic conditions developing after only a few decades. This artificial eutrophication has already happened in many parts of the world including the Norfolk Broads and parts of Holland, Denmark and Norway.

To renew all the water in a lake may take up to a hundred years compared to a few days for the renewal of the water in a river. Consequently, lakes are particularly susceptible to pollution such as artificial eutrophication.

Development of eutrophy in lakes

31

Oligotrophic Lake

Artificial input of nutrients

32

Effects of Eutrophication

• An increase in plant and animal biomass• An increase in growth of rooted plants, e.g. reeds• An Increase in turbidity (cloudiness) of water• An increase in rate of sedimentation • The development of anoxic (anaerobic) conditions (low

oxygen levels)• A decrease in species diversity• A change in dominant biota (e.g. carp replace trout and

blue-green algae replace normal algae) and an increase in the frequency of algal blooms.

33

Eutrophic lake with high concentrations of plant nutrients, especially PO4

Rapid growth of algae/cyanobacteria

34

Heavy algal growth, increased turbidity and increased sedimentation

Increased growth of plants e.g. reeds

35

Consequences of Eutrophication

Some of the main consequences of eutrophication are:

• increased vegetation may impede water flow and the movement of boats

• the water may become unsuitable for drinking even after treatment

• decrease in the amenity value of the water (e.g. it may become unsuitable for water sports such as sailing)

• disappearance of commercially important species (such as trout)

36

Development of anaerobic conditions

Algal blooms followed by lysis

37

Sources of Phosphorus in Lakes

• Weathering of Rocks

• Wastewaters – Industrial – Municipal

• Seepage from Septic Tanks

• Agricultural Runoff (Fertilizers)

Leonard W. Casson, Ph.D., P.E., DEELeonard W. Casson, Ph.D., P.E., DEE

38

Reducing Eutrophication• Treating effluent before it reaches the lake.• reducing the use of phosphates as builders in detergents.• reducing the use of nitrate containing fertilisers.• using tertiary sewage treatment methods to remove

phosphate and nitrate before discharge of the effluent into rivers and lakes.

• directing treated waste water away from lakes to rivers and the sea.

• aerating lakes and reservoirs to prevent oxygen depletion particularly during algal blooms.

• removing phosphate-rich plant material from affected lakes.• removing phosphate-rich sediments by dredging..

39

Effects of Oxygen

• Concentration of oxygen affects microbial growth.

• Redox potential decreases from +600mv (aerated) to –350mv (anaerobic).

• Redox potential depends on pH (above results at neutral pH.

• Effects more severe at acid pH e.g acid mine drainage.

40

Effects of reduction of [O2]

• +600mv Normal redox potential for well aerated water. Large number of aerobic species e.g. Pseudomonas. Alkaligenes, Achromobacter, appendaged bacteria, Aquatic phycomycetes, diatoms (Pinnularia, Navicula), algae e.g. Ankistrodesmus (Chlorophycae).

• +600 to +200mv Facultatively anaerobic Gram-negative rods predominate. NO3

-, Mn, Fe reduced. Pseudomonas, diatoms (Nitzschia, Synedra), filamentous chlorophycae e.g. Stigeoclonium, cyanobacteria e.g.Calothrix.

41

Effects of reduction of [O2]

• +200mv to 0mv Dominance of cyanobacteria, microaerophiles and flagellates.

– Pseudomonas, Oscillatoria, Phormidium, Spirillum, Chlamydomonas, Euglena.

– “Sewage fungus” complex(see later).

– Nitrification begins to be inhibited.

42

Effects of reduction of [O2]

• 0mv to –150mv Nitrification inhibited, build-up of H2 and fermentation products (acids, alcohols), SO4

2- reduced to H2S, Blooms of green and purple sulphur bacteria, population switches to anaerobes.– Desulphovibrio, Clostridium, Chromatium,– Beggiatoa (microaerophilic sulphur oxidisers )

in upper layers.

43

Effects of reduction of [O2]

• -150mv to –350mv terminal stage with CH4, H2, H2S production.

– Devoid of eukaryotes except anaerobic protozoa, Tubifex etc.

– Anaerobes only.– Very few species.

44

Consequences of anaerobiosis

• When plants and algae die their remains gradually sink and are consumed by aerobic bacteria. Microbial predators such as Lysobacter spp. can bloom and kill the algae rapidly. This results in a reduction of the level of dissolved oxygen. Eventually, often near the bottom of a lake, virtually no oxygen remains and the water is said to be anoxic. Under these conditions anaerobic bacteria flourish. Anaerobic bacteria often produce foul-smelling compounds such as:– hydrogen sulphide (H2S) thioalcohols (RSH) and methyl mercaptan

(CH3SH)

– ammonia (NH3) and polyamines such as cadaverine.

resulting in the water becoming extremely unpleasant.

45

Low oxygen concentrations result in fish kills

46

“Sewage fungus”

• Bacteria:– Sphaerotilus natans, zoogleal bacteria, Beggiatoa,

Flavobacterium.

• Filamentous fungi:– Geotrichium, Leptomitus lacteus (Apodya lactea),

Fusarium.

• Algae:– Stigoclonium tenue, Navicula sp., Fragilaria, Synedra,

Cladophora.

47

“Sewage fungus”

• Protozoa:– Colpidium– Childonella– Cinetochilum– Trachellophylum– Paramecium– Uronema– Hemiophrys– Glaucoma– Carchesium

48

Synedra

49

Sewage fungus organisms

Marine Microbiology

51

Oceans

• Primary producers are algae and cyanobacteria = phytoplankton.

• Responsible for 40% total photosynthesis.

• In open ocean cyanobacteria, particularly Prochlorococcus and Synechococcus are dominant (>105 ml-1).

• Large numbers of viruses (107 ml-1) 1030 in total.

52

Viruses may control phytoplankton levels

OCEANS

• Cyanobacteria are the most common type of bacteria in the ocean, and SAR11 (Pelagibacter clade [group of closely related organisms] of the Synechoccus types of bacteria) is the most common organism on earth, accounting for roughly 25% of all the bacteria in the ocean, 50% in the euphotic zone . (Morris et al., 2002, Nature Dec 420:806-810). Globally 2.4 x1028 cells.

• Prochlorococcus and Synechoccus species dominate the microbial ecology of the ocean. They account for 25% of global photosynthesis.

53

54

Oceans

• Phytoplankton fed upon by zooplankton etc.• Productivity in oceans limited (?by

micronutrients e.g. Fe2+).• Open seas have 90% of surface area but

only produce 0.7% of fish cf. coastal zones with 54% (remaining 46% in so-called “upwelling” regions where currents bring nutrient rich sediments to the surface.

55

56

57

58

Marine “snow”

59

60Mid ocean ridges (spreading tectonic plates)

61

62

63

64

65

66

67

68

69

70

Black Smokers

• Organisms

Korarchaeota – Archaea: – Extremophiles– Pompeii worms:

A. pompejana– Gastropod:

Cypraea chinensis

• Crustacea feed on these organisms

71

Symbiosis

• Mussels– Microbes live in the gills, using the chemicals from

the hydrothermal vent fluids to produce sugars– Mussels utilize those sugars

• Clams– Also have bacteria that live in their gills that perform

the same process as the microbes that live symbiotically with the mussels.

72

Tubeworms e.g. Giant tubewormRiftia pachyptila

• Do not have a mouth or a stomach, so they use billions of symbiotic bacteria to produce sugars from carbon dioxide, hydrogen sulfide and oxygen in an organ termed a symbiosome.

• Use some of the sugars as food and provide the bacteria with the hydrogen sulfide and oxygen that the worms take up from the water

73

74

75

Vent crab

• The hydrothermal vent crab, Bythograea thermydron, is a top predator at vent sites in the Pacific Ocean. This crab is present in such high densities that scientists actually use it as an indicator that they are approaching an active vent field.

• The vent crab is typically found among dense clusters of tubeworms at an average depth of 1.7 miles and can tolerate a temperature gradient that ranges from 77°F in the tubeworm clumps, to 36°F, which is the temperature of the water surrounding the vent sites.

76

Vent crab

77

Pompei worm

• The Pompeii worm (Alvinella pompejana) can survive an environment as hot as 80° C

• Covering the Pompeii worm’s back is a “fleece” of thermophilic bacteria.

78

Pompei worm

79

Common copepod

Common limpet

80

gastropod

Glob snail Gyre snail

Maia snail

81

Vent fish

Rattail fish Fathead sculpin

82

Anemone

Ciliate

Nematodes

Palm worm

83

OTHER ANIMALS

84

Some web sites

• www.ocean.udel.edu/expeditions/index.html

• www.pmel.noaa.gov/vents/geology/video.html

• http://www.shef.ac.uk/aps/level2modules/aps201/aps201.html

85

Books

• Biology of freshwater pollution , C.F. Mason. - 3rd ed.. - Harlow : Longman, 1996. - 0582247322

• Microbial ecology : fundamentals and applications, Ronald M. Atlas, Richard. - 4th ed. - Menlo Park, Calif.; Harlow : Benjamin/Cummings, 1998. - 0805306552

• Manual of environmental microbiology / editor-in-chief Christon J. Hurst. - Washington, D.C. : ASM Press, 1997. - 155581087x

• VARNUM, A.H.. - Environmental microbiology. - Manson, 1997. - q5356919

• Freshwater microbiology David Sigee, John Wiley, 2002