International Biodeterioration & Biodegradation, Vol. 40. No.

24 (1997) 247-253
0 1997 Elsevier Science Limited
All rights reserved. Printed in Great Britain
PII:SO964-8305(97)00046-Z 0964~8305/97 $17.00+o.oo
ELSEVIER

Trevi Fountain: An Evaluation of Inhibition Effect

of Water-repellents on Cyanobacteria and Algae

M. P. Nugari & A. M. Pietrini

Istituto Centrale per il Restauro, Rome, Italy

This work presents the biological aspect of a more extensive research to choose a
water-repellent to apply to stone surfaces of the Trevi Fountain in Rome
(Italy).The effect of water-repellent treatments on cyanobacterial and algal
colonizations was investigated and the alteration induced by microflora on
efficacy of the polymers used, was also considered. Carrara marble and travertine
from Tivoli samples were treated with an acrylic polymer, two oligomeric
polysiloxanes and an ethylsilicate polysiloxane. The stone samples were partly
immersed in slow-flowing water inside polyethylene vessels placed in the Trevi
Fountain basin. The qualitative and quantitative characteristics of biocoenosis
were determined. The characteristics of polymers were evaluated by contact
angle, measures of surface roughness, SEM observations and EDAX
microanalysis. The presence of the tested products on stone samples had no
inhibition effect on the microflora growth.
En este trabajo se presentan 10s aspectos biologicos de una investigation mas
amplia cuyo objetivo es la selection de un product0 hidrofugante para aplicar a
las superticies rocosas de la ‘Fontana di Trevi’ (Roma, Italia). Se analizo el efecto
de 10s tratamientos sobre cianobacterias y algas, considerando tambien la
variacibn inducida por la microflora en la eticacia de 10s polimeros empleados.
Se trataron muestras de marrnol de Carrara y travertino de Tivoli con un
polimero acrilico, dos polisiloxanos oligomtricos y un etilsilicato polisiloxano.
Las muestras, colocadas en vasos de polietileno, se dejaron dentro de la taza de la
fuente, en una zona donde el agua discurre lentamente, de modo que quedaron
parcialmente sumergidas. Se determinaron las caracteristicas cuantitativas y
cualitativas de biocenosis. La eficacia de 10s polimeros fue evaluada mediante
medidas del angulo de contact0 y rugosidad superficial, observation con SEM y
microanalisis con EDAX. Asi, se determine que 10s productos ensayados no
presentan un efecto inhibidor de1 crecimiento de la microflora. 0 1997 Elsevier
Science Ltd. All rights reserved

INTRODUCTION importance since the zones of the monument in

contact with water constituted a particularly
Trevi Fountain is a grandiose fountain of the 18th favourable environment for the growth of
century, formed by an architectural facade and a photoautotrophic microorganisms (Ricci &
large basin where a winged chariot of Neptune is Pietrini, 1994). The surfaces of the monument
drawn by two tritons struggling out of the rough- before restoration were in fact colonized by
hewn stone. The fountain is built mainly of Cyanobacteria and algae, which formed thick
travertine from Tivoli (cliff) and Carrara marble patinas of different colours (Pietrini, 1991).
(sculptures). It is very difficult to prevent the formation of
The state of conservation of the fountain these biological alterations (Tiano et al., 1995).
appeared fairly good but a conducted series of There is little research that verities the possible
diagnostic surveys showed the need for restoration inhibition of microbial growth by water repellents.
work. The degradation factors were examined On the basis of this consideration, and owing to
from the chemical, physico-chemical, and the unfavourable environmental conditions, such
biological point of view. as the high level of air pollution (the fountain is
The biological aspect was of particular located in the centre of Rome), the high relative

247
248 hf. P. Nugari, A, M. Pietrini

humidity and continuous contact of some parts of
the monument with water, it was decided to
consider the use of a protecting agent to be
applied on the stone surface at the end of the
restoration work.
The choice of products to be used as water-
repellents was based on an experimental
evaluation in accordance with chemists from the
Istituto Centrale Restauro, Rome, who carried
out a parallel study analysing the chemical and
physico-chemical properties of these substances
and the variations induced by their application on
the substrata (Laurenzi Tabasso et al., 1991).
In particular, the present paper reports some
experimental tests carried out on stone samples to
evaluate the inhibition effect of selected water-
repellents on cyanobacterial and algal
colonization. The alteration induced by microbial
growth on the efficacy of the polymers used was
also considered (Salvadori & Nugari, 1988).
MATERIALS AND METHODS
Four products with water-repellent properties were
applied on test stone slabs (lOxlOx2cm). Two
stone types representative of those used for the
fountain were tested: white Carrara marble and
travertine from Tivoli.
Trade-marks and producers, the chemical
composition, the solutions and the application
methods of the protectives (water-repellents)
tested are reported in Table 1.
Experimental procedure
For the purpose of highlighting the performance of
the tested products from the biological point of
view, the treated samples were exposed in the
environment of the fountain at natural weathering
conditions; the time available was in fact sufficient
(1 year) to verify the behaviour of the products in
the actual environmental conditions of the
monument. The samples were partly immersed in
slow-flowing water in polyethylene vessels and
placed in the basin of the fountain.
The colonization patterns were inspected every 3
weeks by direct observation in situ for the entire
period of testing. The autotrophic microflora
grown both. on the surface of the samples and in
the immersion water, was isolated. Cultural
analyses were carried out according to the Italian
methods Normal (NORMAL 9/88, 1990). A study
of the morphoanatomical characteristics was
performed by optical and scanning electron
microscope (SEM) (Philips 500) to identify the
isolated genera. The samples were prepared
according to the technique described by document
NORMAL 19jS5 (1985).
SEM observations were carried out only on
travertine samples before immersion in water and
after 3 and 12 months of immersion. For technical
reasons, the samples were examined only after
measuring some physico-chemical properties by
analyses (dry weight) that had produced partial
dehydration of microbial cells. Microanalyses
were also made by means of an energy-dispersive
X-ray analyser (EDAX).
The effect of biological growth on the products
was evaluated by measurements of the surface
roughness and of water repellency after 3, 8, and
12 months of ageing.
The roughness profile was recorded using a
Surtronic 3, Taylor and Hobson Prolilograph.
The average roughness ‘Ra’ was evaluated. ‘Ra’
was defined as the average height of the
irregularities measured with respect to a central
line within a prescribed length. In particular, the
following parameters were adopted: exploration
speed = 0.25 mm s-l; pick-up (feeler pins) = 2 mm;
range 99.9mm; in addition, for marble, cut-off
=0.8mm and length of travel =4.5 mm, while for
travertine, cut-off = 2.5 mm and length of travel
= 13mm.

Table 1. Tested Products

Product Chemical composition Concentration And Solvent application method

used
Imlar CPC 1175T (Du Pont) acrylic polymer 33-34% in ready to use brush, two crossed coats
water
Rhodorsil RC80 (Rhone- ethylsilicate polysiloxane 70% ready to use brush until repelled
Poulenc) in white spirit
Wacker 290L (Wacker- oligomeric polysiloxane 80% 1: 11 by weight in Solvesso 100 brush, one coat
Chemie) in Naphta/DAA
Wacker 280 (Wacker-Chemie) oligomeric polysiloxane 100% 1: 11 by weight in Solvesso 100 brush, one coat
 Evaluation qf inhibition effect of water-repellents on Cyanobacteria and algae 249

The water repellency of the treated surfaces was qualitative characteristics of the biocoenosis; the
measured by a contact angle method in dominant types were in fact the same, the only
accordance with NORMAL 33/89 (1991) with a differences regarding taxa were not numerically
Lorentz and Wettre apparatus. significant. As regards quantity, there were signifi-
cantly more photoautotrophic microorganisms on
the stone samples than on the monument; the
RESULTS slow flow of the water in the polyethylene vessels
with respect to the normal conditions of the
The biological analyses showed very similar results fountain functioning was, in fact, a condition
on all treated samples and untreated ones under favouring the biological growth.
test conditions: a considerable growth of SEM observations and EDAX microanalysis
photoautotrophic microflora was detected on the showed that the protecting agents were clearly
immersed surface of samples, while no types of visible only for a short while after the treatment.
biological growth were encountered on non- After 1 year of ageing, on non-immersed areas of
immersed area. stone slabs it was impossible to distinguish the
A total of 13 taxa were identified, four of which siloxanic products, while in the case of acrylic-
were Cyanobacteria, six were Bacillariophyta, and based product, irregularly distributed traces of
three were Chlorophyta. The presence of the resin could be seen (Fig. 1).
different protective agents on test stone slabs did In the areas immersed in water, the observation
not inhibit the Cyanobacteria and/or algal growth. of the products was considerably hampered by the
The taxa and the distribution were very similar on presence of microflora that had grown on the
ail test samples, independently of the presence or surfaces (Fig. 2). The mucilaginous debris of the
of the type of protective product. Only a slightly cellular sheaths of Cyanobacteria concealed the
greater growth was noted on the untreated treated surfaces: their aspect in many cases was
samples. similar to that of the resins and only organic
An analysis of the water in which the samples microanalysis allowed the glycoproteic
were immersed revealed the same types of composition to be confirmed (Fig. 3). Moreover,
microflora as found on the surfaces of stone many siliceous walls of diatoms (Bacillariophyceae)
samples. In addition, the microflora that had were present because the mechanical cleaning to
grown on the test samples and the one that was reduce the biomass did not succeed in removing
identified on the monument prior to the most of them (Fig. 4); on the other hand, more
restoration works were compared (Table 2). This drastic methods would have damaged the water-
comparison revealed a basic homogeneity of the repellents.

Table 2. Presence of Microflora on the Test Samples and on the Monument Before Restoration

Microorganisms Test samples Monument

Cyanobacteria
Chroococcus sp. _ +
Lyngbya sp. + +
Ljvzgbya tenue + _
Microcystis sp. _ +
Oscitfatoria sp. + +
Phormidium sp. + +
Bacillariophyceae
Amphora sp. + _
Eunotia sp. + _
Frustulia sp. + _
Nitzschia sp. + +
Pinnularia sp. + +
Synedra sp. + _
Chlorophyceae
Chlorella sp. + +
Haematococcus sp. + _
Scenedesmus sp. + _
Stichococcus sp. _ -
250 M. P. Nugari, A. M. Pietrini

Fig. 1. Non-immersed area of the travertine treated with Zmlur CPC 1175T, after 1 year of aging; visible traces of resin. SEM
micrograph. (Zanella D.-Sopr. BB.AA.SS. Venice)

Fig. 2. Area immersed in water of the travertine treated with Rhodorsil RC’80; evident signs of autotrophic microflora. SEM
micrograph. (Zanella D.-Sopr. BB.AA.SS. Venice)

In connection with water repellency, it was
found that after 3 months in all polymers, the
contact angle values measured on non-immersed
surfaces of samples were high and similar to that
at time 0, while the values recorded on immersed
areas were significantly decreased. After 8 and 12
months, all the products in immersion lost their
water-repellent properties. These results can be
explained by the rapid degradation of the
products through continuous contact with water
and by the presence on test samples of
mucilaginous sheaths which, once being
dehydrated by the treatment at 60°C as required
by the contact angle method, are very
hydrophilous.
A comparison of the results obtained in the two
different lithotypes indicates that on marble
samples the lost of water repellency was very
similar in the four products, while for travertine
the Wacker 280 showed slightly higher water-
repellent properties than the other products.
Microscopic examinations of stone samples
showed that marble and travertine after biological
colonization presented a grain-like surface (Figs 5
 Evaluation qf inhibition effect of water-repellents on Cvanobacteria and algae 251

Fig. 3. Immersed area of non-treated travertine, after 3 months of ageing: clearly visible are the mucilaginous debris of
extracellular sheats. SEM micrograph. (Zanella D.-Sopr. BB.AA.SS. Venice)

Fig. 4. Siliceous frustules of nennate diatoms on the surface of a travertine test samnle with lmlur CPC /I 75T after mechanical
‘cleaning. SEM micrograph. (Zanella D. - Sopr. BB.AA.SS. Venice)

and 6). This surface appearance was verified from samples had the greater dispersion of data.
roughness measurements. Neverthless the differences encountered were not
The roughness profile, both of marble and significant.
travertine samples, was higher in the immersed
parts than in the non-immersed ones. This could
be due to the residuals of microflora and CONCLUSIONS
calcareous incrustation on the substratum.
A similar roughness was detected on the stone The results of this study prove that the protective
surfaces treated with the three siloxanic products polymers tested, either the three polysiloxanes or
at the different times of ageing, while the the acrylic product, have no inhibition effect on
samples with the acrylic based product had the autotrophic microflora growth. The different
highest roughness after 12 months; the travertine chemical nature of the water-repellents
252 M. P. Nugari, A. M. Pieirini

Fig. 5. Non-treated marble sample after 1 year of ageing: (a) non-immersed area; (b) immersed area. Light micrograph, original
magnification x 25.

conditioned neither the type of biocoenosis nor the
intensity of development.
The degradation of the products immersed in
water was very rapid and the analyses carried out
have not allowed us to understand the different
role of water and of microorganisms in modifying
the physico-chemical characteristics of the
protective agents.
In conclusion, the experiment indicates that all
the protective agents studied had a limited
durability in the environmental conditions of the
Trevi Fountain.
The obtained results are related to a situation of
immersion in water and/or closeness to water, and
they are insufficient to demonstrate the general
ineffectiveness of the products tested in preserving
stone materials from microbial growth; further
tests should be carried out under less stressed
conditions.
The short time of efficacy of the water repellents
tested in this experience leads us to conclude that
the products used for restoration must be
periodically checked in order to plan periodic
maintenance works, where necessary.
ACKNOWLEDGEMENTS
The authors wish to thank Dr 0. Salvadori and D.
Zanella of Soprintendenza ai Beni Artistici e
 Evaluation of inhibition effect of water-repellents on Cyanobacteria and algae 253

Fig. 6. Non-treated travertine sample after 1 year of ageing: (a) non-immersed area; (b) immersed area. Light microgi
original magnification x25.

Storici of Venice for kind collaboration in SEM Pietrini, A. M. (1991) Un’indagine conoscitiva sulla micro-
flora. In Fontana di Trevi-La Storia, il Restaura, pp. 187-
observations and EDAX analyses. 189. Carte Segrete, Rome.
Ricci, S. & Pietrini, A. M. (1994) Caratterizzazione della
microflora algale presente sulla Fontana dei Quattro
Fiumi, Roma. In III International Symposium on the
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