Environ Monit Assess

DOI 10.1007/s10661-009-1180-6

Antimicrobial resistance of heterotrophic marine bacteria

isolated from seawater and sands of recreational beaches
with different organic pollution levels in southeastern
Brazil: evidences of resistance dissemination
Ana Julia Fernandes Cardoso de Oliveira ·
Paula Thais Ranzani de França · Aline Bartelochi Pinto

Received: 18 May 2009 / Accepted: 9 October 2009

© Springer Science + Business Media B.V. 2009

Abstract Antimicrobial resistance of marine het-
erotrophic bacteria to different antimicrobials
agents were evaluated in seawater, dry and wet
sands from three marine recreational beaches with
different pollution levels. In all studied beaches,
the greatest frequencies of resistance were found
in relation to penicillin. On Gonzaguinha, the
most polluted beach, 72.3% of all isolated strains
showed simple resistance, whilst 8.33% had mul-
tiple resistance. The values found on Ilha Porchat
beach, were 70.8% and 6.9% for simple and mul-
tiple resistances, respectively. On Guaraú, the less
polluted beach, only 35.3% of isolated strains
had simple resistance. Multiple resistance was not
observed. While samples from Gonzaguinha and
Ilha Porchat beach showed isolated strains re-
sistant to seven and six different antimicrobial
agents, respectively, samples from Guaraú beach
were resistant only to penicillin and erytromicin.
A. J. Fernandes Cardoso de Oliveira (B) ·
P. T. Ranzani de França · A. B. Pinto
Laboratory of Marine Microbiology,
Campus do Litoral Paulista,
Universidade Estadual Paulista,
Praça Infante Dom Henrique, s/n, 11330–900,
São Vicente, SP, Brazil
e-mail: ajuliaf@clp.unesp.br
The positive correlations obtained between the
degree of seawater contamination and frequency
and variability of bacterial resistance indicate that
polluted marine recreational waters and sands are
sources of resistant bacteria contributing thus, to
the dissemination of bacterial resistance.
Keywords Marine heterotrophic bacteria ·
Resistance · Seawater · Sand beach ·
Contamination
Introduction
Coastal areas are complex formations including
bays, estuaries, beaches and large semi-closed
areas, where there is a high concentration of popu-
lation development. The relation between popula-
tion increases in coastal regions and changes in the
environment has been known since many years.
However, attention has been given only recently
to environmental impacts on marine ecosystem
and on population health caused by the develop-
ment of these areas.
Besides the marine microbiota, seawater and
sediments can contain a significant non-
indigenous microbiota composed by bacteria,
virus and protozoan (some of them are patho-
gens), that are discharged to the environment
in domestic sewage and urban drainage water

(Austin 1988; Sato et al. 2005; Oliveira et al.
2007; Oliveira and Pinhata 2008). The presence
of such microorganisms in recreational seawater,
sediments and beach sands affect the quality of
these habitats, leading to high risk of beachgoer’s
health due to waterborne and other illness, as
well as to the high resistance that can be showed
by these microorganisms to several antimicrobial
agents such as antibiotics (Erdem-Kimiran et al.
2007; Goni-Urriza et al. 2000; Irvensen et al. 2004;
Meirelles-Pereira et al. 2002; Oliveira and Pinhata
2008; Schwartz et al. 2003; Tejedor et al. 2001).
The bacterial resistance to antimicrobial agents
can be intrinsic, associated to the incapacity of
these agents to reach the sites of action in the
cell. In addition, it can be extrinsic, related to
dynamic processes such as changes in antibiotics
transport within the cell, molecular modifications
or enzymatic production that modify and inac-
tivate the antibiotic (Hermansson et al. 1987;
Davies 1994). This resistance can be determined
by both genes from chromosomes as well as by
extra-chromosome genetic elements (R-plasmid;
Gomes-Lus 1998; Hirsch et al. 1999).
Besides their resistance to several antimicro-
bials, bacteria can also, through conjugation with
bacteria from the same or different species,
acquire resistance factors present in plasmid
(Huycke et al. 1998; Kühn et al. 2005). There-
fore, faecal bacteria in domestic sewage with a
high level of antimicrobial resistance can dissem-
inate this resistance to indigenous bacteria. The
frequency and variability of resistant strains of
bacteria to different antimicrobial has increased
in the last time, becoming a great challenge for
the treatment of infectious diseases, which should
be carefully considered (Arvanitidou et al. 2001;
Halling-Sorensen et al. 1998). Therefore, this
study aims to evaluate the resistance of het-
erotrophic bacteria to antimicrobial agents, which
were isolated from seawater and sand samples in
three recreational beaches under different levels
of pollution. In addition, environment potentials
as sources of dissemination of bacterial resistance
were evaluated based on the relationship between
quality of marine recreational waters, bacterial
habitat, frequency and type of resistance.
Environ Monit Assess
Materials and methods
Study areas
São Vicente (23◦ 57 S and 46◦ 23 W) and Peruíbe
(24◦ 19 S and 46◦ 59 W) are located on the Baixada
Santista (“Santos Lowland”), south coast of São
Paulo State (Fig. 1). The Baixada Santista is so
named due to its geography, since it borders the
Serra do Mar (“Sea Highlands”). It occupies an
area of 2,373 km2 , with 161 km2 of beaches. Due to
the heterogeneity of its beaches and the proximity
to urban centres, such as São Paulo city, one of
the most important economical activities in São
Vicente and Peruíbe is tourism.
Peruíbe is located within four conservation
units: Juréia Itatins (considered by UNESCO as
Natural Site on World Heritage), Tupiniquins,
Cananéia-Iguape-Peruíbe and Serra do Mar. It
contains 18 beaches, from which six have their
water quality monitored by São Paulo State Com-
pany for Environmental Sanitation Technology
(CETESB). Since it has a great part of its area
under legal protection and also due to its longer
distance to large cities (e.g. São Paulo), Peruíbe is
still well preserved. Guaraú beach is considered to
have the best sanitary quality among all evaluated
beaches and its water quality is considered to be
excellent over 90% of the time, which is attributed
to its geographical configuration, distance from
city downtown and low rates of urban occupation.
Domestic sewage is collected and directed to a
sewage treatment plain, where it is preconditioned
and submitted to the reduction of organic load
in stabilisation lagoons. After organic load reduc-
tion, effluents are discharged into Preto River,
which runs directly to the sea, in the southern part
of Peruíbe (CETESB 2006).
Five beaches belong to São Vicente:
Itararé, Milionários, Ilha Porchat, Divisa and
Gonzaguinha beach. The last ones are the most
affected by organic contamination, whilst Ilha
Porchat shows the lowest levels of pollution
(CETESB 2008).
The worse bathing water quality at
Gonzaguinha beach is associated with its
geographical configuration and the high inflow of
Environ Monit Assess
BRAZIL
SÃO
PAULO
Fig. 1 Detail of studied areas on Gonzaguinha and Ilha Porchat beaches, located in São Vicente, Baixada Santista (São
Paulo State, Brazil)
organic load brought by the effluents discharged
at Mar Pequeno, São Vicente Channel and
the adjacent hydrographical system, which ulti-
mately reach this beach (CETESB 2006). Since
Gonzaguinha Beach is located in a sheltered
area, dispersal of the microorganisms is limited,
allowing bacteria and other microorganisms to
be accumulated in areas used by beachgoers. On
the other hand, Ilha Porchat Beach is located in
a less sheltered area, where circulation processes
and the constant renewal of the water facilitate
dispersal of the pollutants. These processes
favour the dispersal of microorganism seaward
and provide better bathing water quality.
In São Vicente, there is a sewage treatment
plant in operation, which is responsible for treat-
ing the total collected sewage (56.6%). Part of the
collected sewage is sent to the submarine outfall
of Santos city (CETESB 2006). After passing by
a bar screen to remove solid residuals and after
chlorination, it is released 4 km away from the
coast, in the central part of Santos Bay. During
summer, the treatment of domestic sewage is not
sufficient, since population in these areas can dra-
matically increase, reaching twice the number of
local residents (CETESB 2006). Therefore, there
is an increase of domestic sewage load, affecting
the quality of marine recreational water in the
region (Sato et al. 2005; CETESB 2006).
Besides the increase of domestic sewage dis-
charge in seawaters, the urban water drainage
reaching beaches, litter brought by beachgoers
and tides as well as the presence of animals on
the beach can also be pointed out as noteworthy
problems in the region that are intensified during
the holiday season.

The sampling beaches (Gonzaguinha, Ilha
Porchat and Guaraú) have their water qual-
ity evaluated by Beach Quality Monitoring Pro-
gramme, which is performed by CETESB. The
programme conforms with the Brazilian legisla-
tion on bathing water quality (CONAMA Reso-
lution no. 274/2000). According to the Brazilian
legislation, enterococci density levels higher than
100 UFC 100 mL−1 in more than 20% of five
consecutive samples in sea water or values higher
than 400 UFC 100 mL−1 in one sample are used to
classify beaches as unsuitable for swimming and
bathing.
Sample collection
Seawater and sand samples used for the micro-
biological analyses were collected during autumn
(in the first week of April 2008) on Ilha Porchat
and Gonzaguinha beaches, in São Vicente, and
on Guaraú beach, Peruíbe (in the second week of
April 2008).
Water samples were collected in sterile flasks,
maintained under refrigeration and analysed
within the next 6 h. Sand samples (0–2 cm) were
collected in two distinct zones of the beach: dry
zone—usually not flooded by seawater, corre-
sponding to the most visited area, and wet zone—
corresponding to the area influenced by tides.
Samples comprised 200 g of sand, collected with
sterile spatula, maintained and transported in ster-
ile bags under refrigeration to the laboratory.
Bacterial isolation
Sand
In the laboratory, sand samples were weighted,
diluted in sterile distilled water (1:10) and agitated
twice in a Kline agitator for about 10 min in
order to wash and extract bacteria from samples
(Oliveira and Pinhata 2008). Volumes of 0.1 ml
from sand supernatant were inoculated in Petri
dishes with Marine Agar 2216E culture media
(Difco).
Environ Monit Assess
Water
To isolate heterotrophic bacteria from seawater
samples, 0.1 ml from each sample were inoculated
in Marine Agar 2216E plates and incubated at
37◦ C during 48 h. Afterwards, some pigmented
and non-pigmented colonies (n = 318), from each
agar plate, were selected and pure cultures of het-
erotrophic bacteria were obtained by the streak
plate method using the same culture media.
A previous screening of all the isolated strains,
based on the colony pigmentation, Gram staining
and bacterial morphology was done, before sus-
ceptibility testing, to ensure that strains belong-
ing to different genera and/or species of bacteria.
Among the colonies isolated, 53.4% were non-
pigmented (gram positive rods and cocci and gram
negative cocci) and 46.6% were pigmented (yel-
low, orange and white gram negative rods).
The 318 isolated colonies, both from sand and
water, were transferred and maintained in Marine
Agar 2216E slope until the evaluation.
Statistical analysis
Correlation between data of contamination and
frequency and variability of bacterial resistance
from study sites were calculated using Pearson’s
correlation coefficient (at 5% significance level).
Antimicrobial susceptibility testing
Sensibility tests to antimicrobials were performed
using the agar disc diffusion methods from
Kirby-Bauer, applied to Marine Agar 2216E, ac-
cording to the National Committee for Clinical
Laboratory Standards (NCCLS 2003). Colonies
(318) were tested to the following antimicro-
bials: amoxicillin (AMO) 30 μg, ampicillin (AMP)
10 μg, ciprofloxacin (CIP) 5 μg, chloramphenicol
(CLO) 30 μg, erythromycin (ERI) 15 μg, strep-
tomycin (EST) 10 μg, gentamicin (GEN) 10 μg,
penicillin (PEN) 10 UI, rifampicin (RIF) 30 μg
and vancomycin (VAN) 30 μg.
Environ Monit Assess
Results
Gonzaguinha beach
From 30th March to 27th April 2008,
Gonzaguinha beach was classified as not
recommended for swimming and bathing,
since it showed enterococci density levels higher
than 100 CFU 100 mL−1 in all samples. Based
on annual data of densities of Enterococcos,
obtained by CETESB during 2008, Gonzaguinha
beach was classified as having bad water quality
and Ilha Porchat as regular, according to the
criteria used by the classification of World Health
Organisation.
Analysing all isolated colonies from
Gonzaguinha beach (n = 144), both from water
and sand samples, 72.3% showed resistance to one
or two drugs, 8.3% had multiple resistance and
19.4% were sensitive to all tested antimicrobials
(Table 1).
Bacteria isolated from water samples (n = 48)
showed 93.8% of resistant strains whilst those
isolated from wet (n = 48) and dry sands (n = 48)
showed 83.3% and 64.6% of resistance, respec-
tively.
In water samples, the highest frequency of re-
sistance was related to penicillin (93.8%) and van-
comycin (31.2%), whilst in wet sand, the highest
frequencies found were to penicillin (70.8%), ery-
thromycin (18,8.0%), amoxicillin and vancomycin
(10.4%), rifampicin (8.3%) and ampicillin (4.2%;
Table 2). Among isolated strains from dry sand,
the greatest frequencies of resistance were ob-
served to penicillin (58.3%), vancomycin (12.5%)
and ciprofloxacin (6.2%; Table 2)
Table 1 Percentage of isolated strains resistant to one
or two antimicrobials (simple resistance), and to at least
three antimicrobials (multiple resistance) and their sus-
Strains Gonzaguinha (n = 144)
Number % Number
One or two antimicrobials 104 72.3
At least three antimicrobials 12 8.3
Total of resistant 116 80.6
Susceptibilities 28 19.4
All isolated colonies from samples from
Gonzaguinha beach showed susceptibility to
chloramphenicol, streptomycin and gentamicin
(Table 2).
Ilha Porchat beach
From 30th March to 27th April 2008, Ilha
Porchat beach had densities lower than 100 CFU
100 mL−1 in 60% of analysed samples and
thus, was classified as suitable for swimming and
bathing (CETESB 2008). Based on annual data of
densities of Enterococcos, obtained by CETESB
during 2008, Ilha Porchat was classified as regular,
according to the criteria used by the classification
of World Health Organisation.
Considering all 72 analysed samples (24 for
each substrate), 70.8% showed resistance to at
least one drug and 6.9% had multiple resistance
(resistance to two or more antimicrobials). Sus-
ceptibility to all tested antimicrobials was found in
22.3% of all samples (Table 1). Bacteria isolated
from water samples showed 75.0% of resistant
strains (n = 18) whilst those isolated from wet and
dry sand showed 91.7% (n = 22) and 66.7% (n =
16) of resistant, respectively.
Among isolated resistant strains in seawater,
70.8% were resistant to penicillin, whilst 20.8%
showed resistance to amoxicillin and vancomycin,
and 16.7% to erythromycin (Table 3). In samples
from wet sand, the highest frequency of resis-
tance was also found to be to penicillin (86.4%).
In addition 25% of isolated strains showed re-
sistance to ciprofloxacin, 20.8% to amoxicillin,
and 8.3% to ampicillin. In samples isolated from
ceptibilities, on seawater, wet and dry sand samples from
Gonzaguinha, Ilha Porchat (São Vicente) and Guaraú
(Peruíbe) beaches, Brazil
Ilha Porchat (n = 72) Guaraú (n = 102)
% Number %
51 70.8 36 35.3
5 6.9 0 0
56 77.7 36 35.3
16 22.3 66 64.2
 Environ Monit Assess

Table 2 Percentage of resistant heterotrophic bacteria iso-

lated from seawater and sand samples from Gonzaguinha bathing (CETESB 2008) in all analysed samples.
beach, in relation to antimicrobials (amoxicillin, ampicillin, In fact, this beach had excellent quality during
chloramphenicol, ciprofloxacin, erythromicin, gentamicin, all the year and was classified as good according
penicillin, rifampicin, streptomycin and vancomycin) to the criteria used by the classification of World
Antimicrobial agent % of resistant strains (n = 116) Health Organisation.
Gonzaguinha beach The frequency of resistant strains isolated from
Water Wet sand Dry sand Guaraú was less than the half of those obtained
(45) (40) (31) for bacteria isolated from Gonzaguinha and Ilha
Amoxicillin 0 10.4 0 Porchat beaches. Regarding all isolated colonies
Ampicillin 0 4.20 0 from Guaraú beach (n = 102), both from water
Chloramphenicol 0 0 0
and sand samples, only 35.8% were resistant to
Ciprofloxacin 0 0 6.2
Erytromicin 0 18.8 0
one or two drugs, and 64.2% were sensitive to all
Gentamicin 0 0 0 tested antimicrobials. There were no strains with
Penicillin 93.8 70.8 58.3 multiple resistance (Table 1).
Rifampicin 0 8.3 0 Bacteria isolated from water samples (n = 34)
Streptomycin 0 0 0 showed 55.9% of resistant strains, whilst those
Vancomycin 31.2 10.4 12.5 isolated from wet and dry sands (n = 34 for each
substrate) showed 32.4% and 17.6% of resistance,
respectively.
dry sand, the frequency of resistance found to In water samples, resistance was found to peni-
penicillin and ciproflaxin was 58.3% and 12.5%, cillin (100.0%) and erythromycin (33.3%), whilst
respectively. Moreover, all colonies were sensitive in wet and dry sands the isolated bacteria were
to chloramphenicol, gentamicin, streptomycin and resistant only to penicillin (100.0%; Table 4). All
rifampicin. colonies isolated from water and sands of Guaraú
beach showed susceptibility to other antimicrobial
agents tested.
Guaraú beach

Guaraú beach, from 30th March to 27th April

2008, was classified as suitable for swimming and

Table 3 Percentage of resistant heterotrophic bacteria iso- Table 4 Percentage of resistant heterotrophic bacteria
lated from seawater and sand samples from Ilha Porchat isolated from seawater and sand samples from Guaraú
beach, in relation to antimicrobials (amoxicillin, ampicillin, beach, in relation to antimicrobials (amoxicillin, ampicillin,
chloramphenicol, ciprofloxacin, erythromicin, gentamicin, chloramphenicol, ciprofloxacin, erythromicin, gentamicin,
penicillin, rifampicin, streptomycin and vancomycin) penicillin, rifampicin, streptomycin and vancomycin)
Antimicrobial agent % of resistant strains (n = 56) Antimicrobial agent % of resistant strains (n = 36)
Ilha Porchat beach Guaraú beach
Water Wet sand Dry sand Water Wet sand Dry sand
(18) (22) (16) (19) (11) (6)
Amoxicillin 20.8 20.8 0 Amoxicillin 0 0 0
Ampicillin 0 8.3 0 Ampicillin 0 0 0
Chloramphenicol 0 0 0 Chloramphenicol 0 0 0
Ciprofloxacin 0 25.0 12.5 Ciprofloxacin 0 0 0
Erytromicin 16.7 0 0 Erytromicin 21.0 18.2 33.3
Gentamicin 0 0 0 Gentamicin 0 0 0
Penicillin 70.8 86.4 58.3 Penicillin 79.0 81.8 66.7
Rifampicin 0 0 0 Rifampicin 0 0 0
Streptomycin 0 0 0 Streptomycin 0 0 0
Vancomycin 20.8 0 0 Vancomycin 0 0 0
Environ Monit Assess
Variability of resistance to antimicrobials
Considering the types of antimicrobial agents to
which the isolated strains were resistant, varia-
tions of resistance were observed among bacte-
ria from Ilha Porchat in comparison to the ones
isolated from Gonzaguinha beach and mainly
from Guaraú beach (Table 4). On Gonzaguinha
beach, resistant isolated strains of bacteria showed
resistance to seven antimicrobials: amoxicillin,
ampicillin, ciprofloxacin, erythromycin, penicillin,
rifampicin and vancomycin. On Ilha Porchat
beach, resistance to six antimicrobials was ob-
served among isolated colonies of resistant
bacteria: amoxicillin, ampicillin, ciprofloxacin,
erythromycin, penicillin and vancomycin. Resis-
tance to only two antimicrobial agents, penicillin
and erythromycin was observed for strains iso-
lated from Guaraú beach samples.
Statistical analysis
A positive correlation between the degree of sea-
water contamination and frequency of bacterial
resistance (r = 0.70) and between the degree of
contamination and variability of resistance (r =
0.84) was found for the studied sites.
Discussion
Beaches constitute a considerable part of coastal
ecosystems and they are habitats of extreme
dynamics under great influence of water, land
and wind (Brown and McLanchlan 1990). Recre-
ational beaches located in large urban centres, as
Gonzaguinha and Ilha Porchat beaches, can be
under different anthropogenic pressures (Lobova
et al. 2002), such as discharge of domestic sewage.
The anthropogenic impacts on coastal areas
through inflow of domestic effluents can be eval-
uated by measuring faecal bacteria levels present
in water and sediments, and more recently, by the
occurrence of bacteria resistance to antimicrobial
agents.
The incidence of high percentages of marine
heterotrophic bacteria resistant to antimicro-
bials used for the treatment of human infec-
tious diseases (mainly to penicillin, amoxicillin,
ciprofloxacin and vancomycin), as we found in our
study (except for Guaraú Beach), for both bac-
teria sampled in water and in dry and wet sands,
indicates the contamination by domestic effluents
in two beaches of São Vicente. In addition, the
observed bacterial resistance may be a result of
the entrance of non indigenous bacteria in marine
ecosystems (Goni-Urriza et al. 2000).
Several studies recently showed that beach
sands may act as reservoirs and/or vectors for
a variety of bacteria (Alm et al. 2003; Bonilla
et al. 2007; Mendes et al. 1993; Papadakis et al.
1997; Whitman and Nevers 2003). Sediments gen-
erally contain higher concentrations of bacteria
than water column (Ghinsberg et al. 1994; Oshiro
and Fujioka 1995) due to favourable conditions of
nutrients (Davies et al. 1995; Villar et al. 1999),
shelter against sunlight (Davies-Colley et al. 1999;
Sinton et al. 1994) and protection against proto-
zoan predation (Davies and Bavor 2000; Pianetti
et al. 2004).
The results of this study indicate that, in addi-
tion to their function as a reservoir and vector of
several pathogenic microorganisms, beach sands
may also accumulate indigenous bacteria resis-
tant to several antimicrobial agents. According to
Chandrasekarn et al. (1998) and Tendencia and
De La Pena (2001), high levels of antibiotic resis-
tance in marine bacteria might be a result from
non-indigenous bacteria with antibiotic-resistant
plasmids entering the seawater.
Oliveira and Pinhata (2008) also observed the
presence of resistant bacteria from human and
animal origin in the same beaches. They showed
that Gonzaguinha and Ilha Porchat beaches had
high percentages of resistant enterococci in sand
and water, showing thus a possible transfer of
resistance factors among bacteria in these ecosys-
tems. Such transfer would thus, contribute to
the dissemination of resistance to antimicrobials
agents (Meirelles-Pereira et al. 2002; Irvensen
et al. 2004; Mudryk 2005; Oliveira and Pinhata
2008). In fact, the transfer of resistance plasmid
carrier (Marcinek et al. 1998; Blom et al. 2000)
in marine habitats was documented for several
bacterial species (see Bya et al. 1986; Sandaa 1993;
Smith 1971).
Sands of Gonzaguinha and Ilha Porchat bea-
ches showed the highest frequencies of resistance

in relation to penicillin. However, heterotrophic
bacteria resistant to amoxicillin and ciprofloxacin
could also be observed. According to Gangle
(2005), since the discovery of penicillin in 1928,
there was a great increase on the resistance pat-
terns to this antibiotic. In 1930, only 1% of iso-
lated bacteria strains of hospital samples were
penicillin resistant, whilst in 2005 it increased to
90%. The high levels of resistance to penicillin,
ampicillin and amoxicillin are of immense concern
due to the high variety of pathogenic microorgan-
isms that show this resistance, such as Enterococ-
cus spp., Staphylococcus spp., Streptococcus spp.,
Salmonella spp., Vibrio spp. and Escherichia coli
(Gangle 2005).
The presence of vancomycin-resistant strains
detected in seawater and sand samples on
Gonzaguinha beach, and in seawater samples on
Ilha Porchat beach are of great apprehension,
since this antimicrobial is used on the treatment
of infections caused by Enterococcus spp. and
Staphylococcus spp., known to show multiple re-
sistance to several others antimicrobials (Huycke
et al. 1998). The occurrence of vancomycin-
resistant bacteria is not an isolated case in the
literature. Kühn et al. (2000) already isolated
vancomycin-resistant Enterococcus spp. in super-
ficial water in Sweden and Spain. Therefore, fur-
ther studies are of crucial importance due to the
possible transfer of bacterial resistance factors to
vancomycin.
Gonzaguinha and Ilha Porchat beaches
showed, during the studied period, higher
frequency and variability of resistant strains that
was related to the degree of faecal contamination
in these environments. Similar results were
obtained by Erdem-Kimiran et al. (2007), who
found a direct correlation between pollution
and bacterial resistance to antibiotics in Mamara
Sea and Black Sea waters. Therefore, the high
incidence of resistant strains in these habitats
may be mainly related to the quantity of inflow
organic effluents and might thus, reflect the
high anthropogenic pressure in these marine
ecosystems.
The presence of animals and litter on the
beaches, such as on the Gonzaguinha and Ilha
Porchat, as well as the urban drainage, can also
be considered as important factors collaborating
Environ Monit Assess
to the presence of bacterial resistance, principally
on sands (Sato et al. 2005). These problems are
intensified during holiday season, mostly because
of the increase of population during these days
and the consequent increase in produced litter and
sewage.
Non-point sources of pollution are difficult to
identify, such as the case of urban drainage and
contamination of beaches by animal faeces, sup-
plying surface waters and sands with faecal bacte-
ria (Hagedorn et al. 1999). Nevertheless, in order
to effectively manage the risks from contamina-
tion, it is crucial to identify the different sources
of faecal pollution. This is particularly important,
as bacteria from human sources establish a high
risk to public health due to the possible presence
of pathogenic organisms (Carroll et al. 2005). In
addition, if the faecal source is known, suitable
management actions can be implemented to pre-
vent further contamination to mitigate health risks
(Harwood et al. 2000).
Considering the multiple bacterial resistance to
antimicrobials, the results of this study disagrees
with Mudryk (2005), who found 20% to 30% of
heterotrophic bacteria with multiple resistance on
marine sandy beach. The frequencies found for
both studied areas in São Vicente were lower
and ranged between 5.56% and 8.33%. Although
these frequencies cannot be considered high if
compared to other studies (Arvanitidou et al.
2001), the presence of such strains should not be
underestimated. The continuous contamination of
beaches by domestic sewage and the consequent
addition of resistant strains to several antimicro-
bials to the ecosystem, together with the capacity
of bacteria in transferring resistance to each other
might lead to an increase of the proportion of
bacteria with multiple resistance to antimicrobials
in a short period of time (Miranda and Zemelman
2001).
Since beach sands can contain great densities of
pathogens with high level of resistance to antimi-
crobials and beachgoers tend to stay most of their
time on the sand (Mendes et al. 1993; Papadakis
et al. 1997), the long contact with contaminated
sands, like from Gonzaguinha and Ilha Porchat,
might be a higher risk to the health of people than
the direct contact with seawater (Ghinsberg et al.
1994; Papadakis et al. 1997).
Environ Monit Assess
From both the quantitative and qualitative
point of view, results obtained in this study
demonstrated that marine recreational waters and
sands may be sources of bacterial resistance. It
may thus, contribute to further dissemination of
resistance to antimicrobials, possibly in high lev-
els, which ultimately may render inefficiently the
use of some antimicrobials against infections.
Further studies about the concentration of an-
timicrobial agents and patterns of bacterial resis-
tance in marine habitats are of crucial importance
for the future, as well as those about the transfer
of bacterial resistance in these ecosystems. New
studies should offer factual data to implement
management control to prevent further spread of
resistance and mitigate the current health risks.
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