Diatoms from the Troodos mountains

Troodos_snowscape_Apr19

Back in April, I wrote two posts about the algae from a stream draining a chromite mine in the Troodos mountains in Cyprus (see “Survival of the fittest (1)” and “Survival of the fittest (2)”).  I also planned to write a post about the diatoms growing in the stream but the slide I prepared has been sitting on my desk over the summer whilst I was distracted by other things.  However, I have just started looking at some samples from metal-enriched streams in the northern Pennines and, curious to see whether a Cypriot chromite mine had similar effects, I blew the dust off the slide and slipped it under my microscope.

The principal effect of toxic pollution is to reduce the number of species found and, in this respect, my sample from the outflow of the Hadjipavlou mine outflow was true to form, containing just eight species.  The most abundant of these was Meridion circulare, accounting for one in four of all the cells.  What is more, many of the cells were visibly distorted (see images a., c. and d., in particular, in the plate below).  This is quite a common phenomenon in metal-polluted streams (see “A twist in the tale”) though I have not seen it quite so obviously in Meridion circulare before. My own pet theory is that one of the enzymes involved in laying down the silica cell wall has a metal co-factor that is displaced by heavy metals.

Meridion_circulare_Hadjipavlou_Apr19

Meridion circulare from thepebbles from the stream draining Hadjipavlou chromite mine in the Troodos mountains, Cyprus, March 2019.  Scale bar: 10 micrometres ( = 1/100th of a millimetre).   The photograph at the top of the post shows snow on the Troodos mountains near the mine.

The only other diatom that was at all common in the sample was Hantzschia amphioxys, which also occurred alongside a smaller population of Hantzschia abundans.  I’ve not come across Hantzschia in metal-enriched streams before: it is a species that is most often associated with habitats that are not permanently submerged.  That may be the case at Hadjipavlou but the water that flows from mines comes from groundwater rather than rainfall so would not be subject to the strong seasonal variations that we associate with Mediterranean streams.  It is hard to draw a firm conclusion from a single visit.   Unlike Meridion circulare, however, neither population of Hantzschia showed any obvious distortion, perhaps due to the Hantzschia cells being more heavily silicified than those of Meridion circulare.

The extent to which cellular distortions are obvious does vary between species, as can be seen in “A twist in the tale …”  which compared three different representatives of the same genus in a metal-polluted stream.  I chose the word “obvious” with care as I do think that these phenomena are more easily seen in long thin cells than in shorter ones.  In the same Pennine streams where distorted Fragilaria are common, for example, I can also see distorted cells of smaller diatoms such as Achnanthidium minutissimum.  But you need a keen eye to spot these reliably.   Some other people have used fluorescent stains to look at other cellular irregularities, such as the position of the nucleus and damage to the nuclear membrane, but these require specialist approaches whereas distortions to cell outlines can be spotted from a standard analysis.

Hantzschia_spp_Hajipavlou_Apr19

Hantzschia abundans (k., l.) and Hantzschia amphioxys (m. – p.) in the from the stream draining Hadjipavlou chromite mine in the Troodos mountains, Cyprus, March 2019.  Scale bar: 10 micrometres ( = 1/100th of a millimetre). 

A few years ago I was involved in a study of diatoms from streams in Cyprus and I dug out some of these data in order to put the Hadjipavlou sample into context.  One immediate surprise was that many of the “reference” (i.e. pristine or near-pristine) samples in that survey also had relatively low diversity.   The 45 samples in this subset had, on average, nine species, and a mean Shannon diversity index of 1.7, compared to eight species and a Shannon diversity index of 1.42 for the Hadjipavlou sample.   I’ve never been a fan of diversity indices as measures of ecological quality (see “Baffled by the benthos (2) and links therein”) although I suspect that average diversity at Hadjipavlou measured over a period of time will always be low whereas average diversity at unimpacted sites is more likely to fluctuate. Equally, low diversity coupled with a second strand of evidence, such as distorted valves, is a useful sign to an ecologist that something untoward is happening.

diversity_indices

Number of taxa (left) and Shannon diversity (right) recorded in 45 samples from “reference” sites (i.e. minimal evidence of anthropogenic alteration) in Cyprus.  The arrows indicate the location of the Hadjipavlou stream within this dataset. 

The irony of writing about a heavily-polluted stream in the Troodos mountains is that the geological conditions which created the metal-rich veins hereabouts also create conditions for many plants endemic to Cyprus.   The serpentine and other ultramafic rocks create metal-rich soils within which few plants can survive (more about these here. I suspect that few of the plant enthusiasts drawn to Cyprus will ever cast more than a cursory glance at the green flocs adorning the abandoned mines of the Troodos mountains.

References

Licursi, M., & Gómez, N. (2013). Short-term toxicity of hexavalent-chromium to epipsammic diatoms of a microtidal estuary (Río de la Plata): Responses from the individual cell to the community structure. Aquatic Toxicology 134-135: 82-91.  https://doi.org/10.1016/j.aquatox.2013.03.007

Survival of the fittest (2) …

As well as the bright green flocs of Tribonema, the stream draining the Hadjipavlou chromite mine also had bright orange-red growths on some of the pebbles on its bed.  These seemed to be composed primarily of the Cyanobacterium Chamaesiphonthough I am still not sure what species.   Using the limited literature I have, from the UK and Germany, I would opt for either Chamaesiphon polymorphusor C. polonicus.   This particular alga was very easy to remove from stones, compared to other epilithic Chamaesiphon species (see “A bigger splash …”).  This is a feature of C. polymorphus, though the colour is more typical of C. polonicus*.  On the other hand, that bright colour could be the response to high solar radiation, so maybe my north European guides are not that reliable.  It could be something else altogether.

Chamaesiphon_polonicus_Troodos

Chamaesiphon growths on pebbles in the stream draining Hadjipavlou chromite mine in the Troodos mountains, Cyprus, March 2019.

Chamaesiphon_Troodos_Mar19

Colonies of Chamaesiphon from Hadjipavlou chromite mine under the microscope.   Scale bar: 10 micrometres (= 1/100thof a millimetre). 

In addition to the Chamaesiphon, there were a few diatoms, mostly Achnanthidium minutissimumand Meridion circulare.   These are typical species of metal-rich streams, as is the general lack of diversity that was evident.   There were also a few filaments of the cyanobacterium Phormidium, along with quite a few Paramecium and Vorticella.  As these are both heterotrophs that feed on organic matter, their abundance is probably at least partly a reflection of the long time that the sample spent in my suitcase between collection and analysis.  The latter is a fascinating organism to watch: it is a goblet-shaped cell with a fringe of cilia around the lip (or “peristome”).  These beat in unison to create water currents that draw small particles towards the cell.   These particles mostly at least an order of magnitude smaller than the algae)  are then collected in food vacuoles where they are digested.   A few of these vacuoles can be seen in the image of Vorticella below.

Vorticella is attached to its substrate by a stalk which contains contractile filaments, giving it spring-like qualities.  Watching a Vorticella is a beguiling experience, with the undulating rows of cilia drawing food into the vestibule (as the opening is known).  At intervals, the whole cell lurched across the field of view as the “spring” in the stalk suddenly contracted, shortening the stalk.  After this, the stalk would gradually extend again, the cilia not having missed a beat meanwhile.   This process may simply be a device that enables the Vorticellato exploit its locality to the full, as well as creating some additional turbulence to keep a steady flow of particles towards the peristome.  To be honest, I haven’t seen a more convincing explanation but, even if we don’t know why it does what it does, Vorticella is a fascinating organism to watch, whether or not I understand what is going on.

I’ll be coming back to talk more about the diatoms in a future post, and writing these posts has also reminded me that I’ve never written about the interesting mine sites almost on my own doorstep.  I cut my ecological teeth looking at these habitats back in the 1980s and they are striking examples of natural selection in action.   So, plenty of potential for more left-field natural history …

Hadjipavlou_organisims_Mar19

Other organisms present in the Hadjipavlou chromite mine. a. – d.: Meridion circulare; e. Phormidiumsp.; f. Vorticellasp.   Scale bar: 10 micrometres (= 1/100thof a millimetre). 

* Note: after I had written this post Brian Whitton confirmed that it was, most likely, Chamaesiphon polonicus.

Survival of the fittest (1) …

Hadjipavlou_mine_March19

When I signed up to a trip to Cyprus in late March I was anticipating feeling some warm Mediterranean sun on my skin after the ravages of the British winter.  I did not expect snow and sleet.   However, as one of our destinations was the Troodos mountains, the location of Cyprus’ only ski resort, maybe it was a case of unrealistic expectations.   Fortunately, we realised our mistake just in time to pack some warm clothes, and the unseasonable weather did, at least, mean that the spring flowers at lower altitudes were, even by Cypriot standards, particularly impressive.

I was in Cyprus primarily as a camp follower on a reconnaissance trip for a geology and botany excursion next year.   Cyprus is, to put it in layman’s terms, the outcome of a collision between the African and European continental plates.   The Troodos mountains are a geologist’s paradise, having a wide range of features arising from this and from associated volcanic activity.   As the molten rocks cooled, minerals precipitate out and the resulting geological strata reflect differences in the melting points of these minerals.   Some of these minerals, such as chromite, are commercially valuable and have been mined for centuries.   Indeed, the name Cyprus itself is derived from cuprous, the Greek word for copper, which was mined here since 4000 BC.

The Hadjipavlou mine is set amidst pine forests close to the highest point of the Troodos.  It was an active chromite mine from 1950 to 1954 and from 1979 to 1982 but was abandoned when cheaper sources of chromite became available in South Africa.   Over a million tonnes of ore were extracted in the period when the chromite mines in the area were active, but now there are few obvious signs apart from this adit driven into the hillside.   A small stream bearing water that has percolated through the rocks and collected in the mine’s galleries emerges from the mine entrance and tumbles down the hillside to join the stream below.   This, on closer inspection, has some quite interesting microbial growths.

First of all, having been told that this is a chromite mine, you might expect the water to carry toxic concentrations of heavy metals.   So you might also be surprised to see abundant growths of bright green algae thriving in the stream immediately downstream of the mine entrance.   This is, in fact, a common phenomenon in mine waters and happens, we think, because the fast-growing algae evolve metal tolerance whilst the animals that feed on them are slower to adapt.   This is, literally, survival of the fittest and, with nothing to eat them, the algae grow prolifically.

These filaments belong to the genus Tribonemawhich, despite being bright green in colour, actually belongs to the yellow-green algae, the Xanthophyta, rather than to the green algae.  This group is actually more closely related to the diatoms than to the green algae, though it can be hard to understand why simply by peering through a microscope.  One useful test is to add a little iodine  solutionto the slide: this binds to the starch inside green algae cells, staining them a dark brown colour.   The Xanthophyta, by contrast, do not have starch as their storage product so the cells are not stained by iodine.   The only other member of this group that I have discussed in this blog is Vaucheria, a very different alga (see “Who do you think you are?”).

Tribonema_affine_Troodos

Tribonema cf affinein the channel draining the Hadjipavlou chromite mine in the Troodos mountains, Cyprus, March 2019.   a. close-up of the alga in situ; b.  microscopic view of filaments; c. fragments of disintegrated filaments showing the H-shaped cell endings.  Scale bar: 10 micrometres (= 100thof a millimetre).   

Tribonemahas simple, unbranched filaments with two or more plate-like chloroplasts arranged around the cell periphery.   One other feature is the arrangement of the cell wall, which tends to consist of two overlapping halves.  When filaments disintegrate (as they often do) the fragments have an H-shape, with each end forming half the cell wall of a different cell.   The cells are, in fact, cylindrical but this is not apparent with the flattened perspective of a high magnification objective.   This feature is not universal in the Xanthophyta, nor is it unique to this group (a few green filamentous algae show the same characteristic) but it is a useful hint that you may be looking at Tribonema.

Whilst lush growths of algae is a common feature of streams draining mines, the species that form these growths can vary.   In the northern Pennines, I am used to seeing green algae in these habitats, but there are at least three different genera that I find.  Typically there is just one filamentous alga in this location, and they tend to be  constant over time: they are reliable sources for material for undergraduate practical classes as a result.  There is more to this story but I will have to come back to it at some point in the future.  .

There is also more to the algal flora of the Hadjipavlou chromite mine but, again, that will have to wait for another post.  I should also confess that, although I visited the mine briefly last year, these samples were collected by Heather, whilst I was sitting snugly below the snow line.