ArticlePDF Available

Preliminary evidence for human fecal contamination in corals of the Florida Keys, USA

Authors:

Abstract and Figures

Corals and reef environments are under increased stress from anthropogenic activities, particularly those in the vicinity of heavily populated areas such as the Florida Keys. The potential adverse impacts of wastewater can affect both the environment and human health; however, because of the high decay rate of bacterial indicators in coral reef waters it has been difficult to document the presence of microbial contaminants and to assign risks in these environments. Here we show initial evidence that microorganisms associated with human feces are concentrated along the surface of coral heads relative to the overlying water column in the Florida Keys. Bacterial indicators (fecal coliform bacteria, enterococci or Clostridium perfringens) were detected in 66.7% of the coral surface microlayer (CSM) samples at levels between five and 1000 CFU/100 ml, but were found infrequently and at low numbers in the overlying water column ( < or = 2.5 CFU/100 ml). Similarly, enterovirus nucleic acid sequences, an indicator of human-specific waste, were detected in 93.3% of the CSM samples and only once in the water column by cell culture. Results show that coral mucus may accumulate enteric microorganisms in reef environments, and may indicate a risk to public and environmental health despite low indicator levels in the surrounding water.
Content may be subject to copyright.
Preliminary evidence for human fecal contamination in corals of
the Florida Keys, USA
Erin K. Lipp
a,*
, Jennifer L. Jarrell
a,1
, Dale W. Griffin
a,2
, Jerzy Lukasik
b
,
Jennifer Jacukiewicz
a
, Joan B. Rose
a
a
College of Marine Science, University of South Florida, 140, 7th Ave. S, St. Petersburg, FL 33701, USA
b
Biosecure, 4641 W 6th St. Suite A, Gainesville, FL 32609, USA
Abstract
Corals and reef environments are under increased stress from anthropogenic activities, particularly those in the vicinity of heavily
populated areas such as the Florida Keys. The potential adverse impacts of wastewater can affect both the environment and human
health; however, because of the high decay rate of bacterial indicators in coral reef waters it has been difficult to document the
presence of microbial contaminants and to assign risks in these environments. Here we show initial evidence that microorganisms
associated with human feces are concentrated along the surface of coral heads relative to the overlying water column in the Florida
Keys. Bacterial indicators (fecal coliform bacteria, enterococci or Clostridium perfringens) were detected in 66.7% of the coral surface
microlayer (CSM) samples at levels between five and 1000 CFU/100 ml, but were found infrequently and at low numbers in the
overlying water column ( 62.5 CFU/100 ml). Similarly, enterovirus nucleic acid sequences, an indicator of human-specific waste,
were detected in 93.3% of the CSM samples and only once in the water column by cell culture. Results show that coral mucus may
accumulate enteric microorganisms in reef environments, and may indicate a risk to public and environmental health despite low
indicator levels in the surrounding water. Ó2002 Elsevier Science Ltd. All rights reserved.
Keywords: Pollution; Corals; Enteroviruses; Indicator bacteria; Florida Keys; Coral mucus; RT-PCR
1. Introduction
Corals and reef environments are under increased
stress from anthropogenic activities, particularly those
in the vicinity of heavily populated areas such as the
Florida Keys. Researchers have noted a loss of diversity
in corals and an increase in diseased or damaged corals
throughout the Florida Keys National Marine Sanc-
tuary (Dustan and Halas, 1987; Harvell et al., 1999;
Porter et al., 1999). In a recent review of coral disease by
Green and Bruckner (2000), the authors noted that 97%
of coral disease in the Caribbean region was docu-
mented in reefs moderately to highly impacted by
human activities. Risks to public health may also be an
issue in these areas. Recently, human waste has been
found to contribute to a high prevalence of enteric vi-
ruses in nearshore waters and canals of the Florida
Keys, and a risk to swimmers in Key West has been
documented (Griffin et al., 1999; Nobles et al., 2000).
Despite this, conclusive evidence that wastewater is
reaching and adversely impacting Keys’ reef environ-
ments and corals is lacking and the topic remains con-
troversial. The potential adverse impacts of wastewater
affect both the environment and human health; however,
because of the high decay rate of bacterial indicators in
coral reef waters it has been difficult to document the
presence of microbial contaminants and to assign risks.
Previous studies have shown that fecal bacteria
and viruses from human waste do not survive long in
saline, warm and highly transparent waters generally
found in reef and other marine environments (Solic and
Krstuvolic, 1992). Yet in marine sediment, enteric
*
Corresponding author. Present address: Department of Environ-
mental Health Science, University of Georgia, 206 Environmental
Health Science Building, Athens, GA 30602, USA. Tel.: +1 706 542
2454; fax: +1 706 542 7472.
E-mail address: lipp@umbi.umd.edu (E.K. Lipp).
1
Present address: North Inlet––Winyah Bay National Estuarine
Research Reserve, Georgetown, SC 29442, USA.
2
Present address: United States Geological Survey, Center for
Coastal Geology and Regional Marine Studies, St. Petersburg, FL
33701, USA.
0025-326X/02/$ - see front matter Ó2002 Elsevier Science Ltd. All rights reserved.
PII: S0 0 2 5 - 3 2 6 X ( 0 1 ) 0 0 3 3 2 - 0
www.elsevier.com/locate/marpolbul
Marine Pollution Bulletin 44 (2002) 666–670
microorganisms have been detected at significantly ele-
vated levels relative to the overlying water column (Lipp
et al., 2001a). A similar accumulation and enhanced
survival may also occur at the coral surface, where
overlying water column conditions may promote rapid
die-off of bacteria and viruses from wastewater. Indeed,
high levels of native bacteria, relative to the overlying
water column, have been noted (Paul et al., 1986), and
bacteria may be chemotactically attracted to mucus
(Ducklow and Mitchell, 1979). Thus, we hypothesized
that the surface microlayer of coral heads (mucus) might
also accumulate microbial indicators of waste and
human viruses, and could thereby provide more direct
evidence for human impacts on reef environments. To
our knowledge this is the first report on the detection of
fecal indicators and human enteric viruses concentrated
in coral surface microlayers (CSM).
2. Materials and methods
One-liter grab samples of surface water were collected
from four sites, between Long Key and Marathon, in
the Florida Keys National Marine Sanctuary in March
2000 (Fig. 1). At each site, a large volume of surface
water (100 l) was also concentrated by adsorption/elu-
tion for cell culture analysis of enteroviruses (Lipp et al.,
2001b; USEPA, 1994). In addition, surface microlayers
(mucus) from 15 corals were sampled by gentle aspira-
tion with a sterile 60 ml syringe. Corals included Mon-
tastraea annularis complex, Siderastraea radians,S.
sideraea and Solenastraea bournouni. Physical and
chemical conditions observed at each site are listed in
Table 2.
Indicator bacteria were enumerated by membrane
filtration and growth on selective media. Duplicate
aliquots of water and mucus were vortexed and filtered
onto 47 mm, 0.45 lm pore size, filters and placed on
appropriate selective media. For Clostridium perfringens,
filters were incubated on mCP medium (AcuMedia,
Baltimore, MD) at 45 °C for 24 h. Yellow colonies that
turned pink upon exposure to ammonium hydroxide
fumes (30 s) were counted (Bisson and Cabelli, 1979).
Filters were incubated on mEI medium at 41 °C for 24 h
and colonies with a blue halo were counted as entero-
cocci (USEPA, 1997). Fecal coliform bacteria were
enumerated on mFC medium; blue colonies were coun-
ted after incubating at 44.5 °C for 24 h (APHA, 1992).
Coliphage were detected and enumerated using the agar
overlay method of Adams (1959), with an Escherichia
coli host (ATCC strain 15597). Five replicate plates of 2
ml aliquots were assayed for each sample and plaques
were counted after 18–24 h incubation at 37 °C.
To assay for infectious enteroviruses in the water
column, concentrated samples were inoculated onto
BGM (Buffalo Green Monkey kidney) cells and exam-
ined for cytopathogenic effects for six weeks (repre-
senting three passages). CSM samples were tested for
enteroviruses using RT-PCR followed by internal probe
hybridization. Viral RNA was extracted using the
RNeasy Kit from Qiagen (Valencia, CA) and 10 llof
purified RNA was used in RT-PCR according to the
conditions described by De Leon et al. (1990). The tar-
get 197 bp amplicon was confirmed by dot-blot hy-
bridization with a biotin-labeled probe internal to this
region, and detected by chemiluminescence (Griffin
et al., 1999, 2000; Table 1). Poliovirus and echovirus
were used as positive controls.
Fig. 1. Map of sampling stations in the Florida Keys. The dotted line box in the inset map of Florida shows the boundary of the Florida Keys
National Marine Sanctuary. The arrow shows the direction of the Florida Current as it passes to the south and east of the Florida Keys. Legend: Key
22––Channel 2; Key 23––100 m off Long Key, Florida Keys Marine Lab; Key 24––Marathon Government Center Boat Basin; Key 25––Long
Key, beach at Florida Keys Marine Lab.
E.K. Lipp et al. / Marine Pollution Bulletin 44 (2002) 666–670 667
3. Results
Fecal coliform bacteria and enterococci were detected
only once in the water column (at Key 24) at 0.5 and 2.5
CFU/100 ml, respectively, while C. perfringens were
never found (Table 2). In contrast, all of the bacterial
indicators were detected at least once from CSM sam-
ples collected from three of the four sites (Key 22, 24,
25) (Table 3). Furthermore, nine of the 15 coral heads
sampled were positive for one or more of the fecal in-
dicators (Table 3). The highest indicator levels were
found in CSM samples from Key 24 with an average of
74 CFU fecal coliform bacteria, 252 CFU enterococci
and 33 CFU C. perfringens per 100 ml. Coliphage were
never detected in water or CSM samples.
Infectious enteroviruses were detected in the water
column only at Key 24 (Table 2), but enterovirus se-
quences were found in CSM samples from all stations
and 93.3% of the mucus samples (14/15) (Table 3, Fig.
2). In 40% of the cases, a strong signal was observed
(Fig. 2). Most positive samples were collected from M.
annularis complex, at Key 22, where the average depth
Table 1
Primer and probe sequences for the specific detection of enteroviruses by RT-PCR (De Leon et al., 1990; Griffin et al., 1999, 2000)
Target Sequence Product size (bp)
50untranslated region (highly conserved) Primer 50-CCTCCGGCCCCTGAATG-30197
Primer 50-ACCGGATGGCCAATC-30
Probe 50-TACTTTGGGTGTCCGTGTTTC-30
Table 2
Physical and chemical conditions, and enteric microorganism in the water column overlying selected patch reefs
Site °C Salinity pH Tidal
stagea
Fecal coliform
bacteria
(CFU/100 ml)
Enterococci
(CFU/100
ml)
Clostridium
perfringens
(CFU/100 ml)
Coliphage
(PFU/100 ml)
Enteroviruses
(MPN/100 l)
Key 22 (Channel 2) 23.9 32 8.1 Flood <0.5 <0.5 <0.5 <10 <1
Key 23 (100 m off Long
Key Marine Lab)
24.7 35 8.0 Flood <0.5 <0.5 <0.5 <10 <1
Key 24 (Marathon Gov’t
Boat Basin)
28.0 35 7.7 Flood 0.5 2.5 <0.5 <10 1.13
Key 25 (beach at Long
Key Marine Lab)
26.0 35 7.9 Flood <0.5 <0.5 <0.5 <10 <1
Samples were collected between 11:00 a.m. and 3:15 p.m., over a period of two days.
a
Tide data were obtained from Tides and Currents Pro software v2.5b (Nautical Software, Beaverton, OR).
Table 3
Enteric microorganisms in CSM samples (CFU or PFU/100 ml)
Site ID Species Fecal coliform
bacteria
Entero-
cocci
Clostridium
perfringens
Coliphage Amplified enterovi-
rus sequence
Key 22 M1 Montastraea annularis complex <5<5<5<10 þþþ
M2 Montastraea annularis complex <5<530 <10 þþ
M3 Montastraea annularis complex <5<55 <10 þþþ
X<5<5 11.7 <10
Key 23 M4 Siderastraea sideria <5<5<5<10 þ
M5 Solenastraea bournouni <5<5<5<10 þþ
X<5<5<5<10
Key 24 M6 Siderastraea radians 60 80 10 <10 þ
M7 Siderastraea radians 70 55 15 <10 þ
M8 Siderastraea radians 115 1000 20 <10 þ
M9 Siderastraea radians 50 50 90 <10 þ
M10 Siderastraea radians 75 75 30 <10 þþ
X 74 252 33 <10
Key 25 M11 Siderastraea radians <5<5<5<10 þ
M12 Siderastraea radians <5<5<5<10 þ
M13 Siderastraea radians <5<5<5<10
M14 Siderastraea radians <5<525 <10 þ
M15 Siderastraea radians <565<5<10 þþ
X<5135<10
X¼mean concentration in CSM per site).
668 E.K. Lipp et al. / Marine Pollution Bulletin 44 (2002) 666–670
of sampled coral heads was 1.5 m. Strong positive
samples were also found at Key 23 from S. bournouni
(collected at 1 m). Only one strong positive was noted
from Siderea radians samples collected at Key 24, where
the average depth was 0.25 m.
4. Discussion
Communities in the Florida Keys rely almost exclu-
sively on on-site wastewater disposal (Paul et al., 2000).
There are at least 24,000 septic systems and 5000–10,000
illegal cesspools, which are underlain by a porous
limestone substrate (Paul et al., 2000; Shinn et al., 1994).
Previous work has traced the migration of wastewater in
septic tanks and shallow injection wells to nearshore
environments throughout the Florida Keys (Griffin
et al., 1999; La Pointe et al., 1990; Paul et al., 1995, 1997,
2000). In particular, areas sampled at Long Key (Key 23
and 25) and Marathon (Key 24) have been shown to
receive wastewater from septic systems on those res-
pective islands, based on viral tracers studies (Paul et al.,
1997, 2000). Despite tracer evidence for wastewater
migration, it has been difficult to routinely document
contamination because indicators rapidly die off in these
warm, saline and highly transparent waters (Solic and
Krstuvolic, 1992). Here we have shown that relative to
the overlying water column, coral mucus can accumu-
late enteric bacteria and viruses in near shore patch reefs
of the Florida Keys that are influenced by poor waste-
water treatment and disposal practices (e.g., septic sys-
tems and shallow injection wells).
While bacterial indicators, in general, were found at
higher levels in the CSM, we observed differences in the
frequency of detection of certain indicators. Clostridium
perfringens was the most prevalent fecal indicator in
CSM samples but was never recovered from the water
column. We speculate that this disparity could have
been due to low oxygen conditions on some of the coral
heads, which might select for this anaerobic bacterium;
however D.O. was not measured. The ability of the
species to form spores might also have provided for
better survival. The absence of coliphage in all samples
was most likely related to a low tolerance for warm
saline waters; MS2 coliphage survives <36 h at 32and
30 °C (McLaughlin, 2000). Coliphage were also isolated
infrequently in previous studies in the Florida Keys
despite a high prevalence of enteric viruses (Griffin et al.,
1999).
Enteroviruses, which specifically indicate contami-
nation with human waste, were detected frequently
throughout the CSM samples, while only one water
column sample was positive. There was no correlation
between the presence of enteroviruses and indicator
bacteria; however, indicator bacteria are known to be
inadequate proxies for enteric viruses (e.g., Bitton
et al., 1983; Griffin et al., 2001; Havelaar et al., 1993;
Lipp et al., 2001a). Patterns of enterovirus detection in
coral mucus noted here suggest that there might be an
association between viral accumulation and specific
coral species and/or sample location and depth. Strong
virus signal was most common in samples from M.
annularis complex and/or those collected at greater
depths. Rohwer et al. (2001) suggest that specific bac-
teria-coral associations exist and, thus, it is possible
that the native microbial community between species
could differentially influence the stability of enteric vi-
ruses that might be accumulated in coral mucus. Fur-
thermore, differences in mucus composition could have
also affected RT-PCR inhibition between sites and/or
species. Alternatively, samples collected at greater
depths tended to produce greater hybridization signal
intensity. Therefore, light attenuation could have
played a role in maintaining enteric viruses, and viral
nucleic acids, in CSM where they could be further
protected from photodamage (Johnson et al., 1997;
Lyons et al., 1998).
5. Conclusions
In this exploratory study we have shown that micro-
bial contaminants associated with human wastewater
can be isolated more frequently and at higher concen-
trations in coral mucus than in the overlying water
column in nearshore areas of the Florida Keys. The
CSM might offer a better record of fecal contamination
in reef areas where fecal indicators and pathogens are
otherwise difficult to detect. Evidence of human entero-
viruses also suggests a public health risk in these envi-
ronments, in the absence of detectable levels of fecal
indicators in the water column. Ongoing and future
work will explore changes in enteric bacterial and viral
levels over time and space, and will assess potential
wastewater impacts in offshore reef areas in both rec-
reational and protected areas. Should similar observa-
tions be made along offshore reefs, sampling of coral
mucus for enteric bacteria and viruses may offer a means
of early and rapid detection of wastewater pollution
and, eventually, might be used to determine risk to coral
and reef health, as well as human health.
Fig. 2. Dot-blot hybridization for RT-PCR amplified enterovirus se-
quences detected by chemiluminescence. Row A, columns 1–12: M1–
M12. Row B, columns 1–3: M13–M15; columns 4, 5 and 6: poliovirus
positive control, echovirus positive control, negative control, respec-
tively.
E.K. Lipp et al. / Marine Pollution Bulletin 44 (2002) 666–670 669
Acknowledgements
Corals were sampled in the Florida Keys National
Marine Sanctuary under permit # FKNMS-2000-010.
We would like to thank Walter Jaap and Jennifer
Wheaton of the Florida Marine Research Institute,
Florida Wildlife Conservation Commission (FWCC) for
logistical support. We are also grateful to Dave Eaken
and John Dotten (FWCC) for sampling support and to
Eugene Shinn (USGS) for careful review of this manu-
script.
References
Adams, M.H., 1959. Bacteriophages. Interscience Publications, New
York.
American Public Health Association, 1992. Standard Methods for the
Evaluation of Water and Wastewater, 18th ed. Washington, DC.
Bisson, J.W., Cabelli, V.J., 1979. Membrane filtration enumeration
method for Clostridium perfringens. Appl. Env. Microbiol. 37,
55–66.
Bitton, G., Farrah, S.R., Ruskin, R.H., Butner, J., Chou, Y.J., 1983.
Survival of pathogenic and indicator microorganisms in ground
water. Ground Water 21, 405–410.
De Leon, R., Sheih, Y.-S.C., Baric, R.S., Sobsey, M.D., 1990.
Detection of enteroviruses and hepatitis A virus in environmental
samples by gene probes and polymerase chain reaction. In: Proceed-
ings of the Water Quality Conference San Diego, CA. American
Water Works Association, vol. 18, pp. 833–853.
Ducklow, H.W., Mitchell, R., 1979. Bacterial populations and
adaptations in the mucus layer on living corals. Limn. Ocean. 24,
715–725.
Dustan, P., Halas, J.C., 1987. Changes in the reef-coral community of
Carysfort Reef, Key Largo, Florida: 1974–1982. Coral Reefs 6,
91–106.
Green, E.P., Bruckner, A.W., 2000. The significance of coral disease
epizootiology for coral reef conservation. Biol. Conserv. 96,
347–361.
Griffin, D.W., Gibson, C.J., Lipp, E.K., Riley, K., Paul, J.H.,
Rose, J.B., 1999. Detection of viral pathogens by reverse transcrip-
tase PCR and of microbial indicators by standard methods in
the canals of the Florida Keys. Appl. Env. Microbiol. 65, 4118–
4125.
Griffin, D.W., Gibson, C.J., Lipp, E.K., Riley, K., Paul, J.H., Rose,
J.B., 2000. Detection of viral pathogens by reverse transcrip-
tase PCR and of microbial indicators by standard methods in the
canals of the Florida Keys (Erratum). Appl. Env. Microbiol. 66,
876.
Griffin, D.W., Lipp, E.K., McLaughlin, M.R., Rose, J.B., 2001.
Marine recreation and public health microbiology: quest for the
ideal indicator. Bioscience 51, 817–825.
Harvell, C.D., Kim, K., Burkholder, J.M., Colwell, R.R., Epstein,
P.R., Grimes, J., Hofman, E.E., Lipp, E.K., Osterhaus, A.D.M.E.,
Overstreet, R., Porter, J.W., Smith, G.W., Vasta, G., 1999.
Diseases in the ocean: Emerging pathogens, climate links, and
anthropogenic factors. Science 285, 1505–1510.
Havelaar, A.H., Olphen, M.V., Drost, Y.C., 1993. F-specific RNA
bacteriophages are adequate model organisms for enteric viruses in
fresh water. Appl. Env. Microbiol. 59, 2956–2962.
Johnson, D.C., Enriquez, C.E., Pepper, I.L., Davis, T.L., Gerba, C.P.,
Rose, J.B., 1997. Survival of Giardia,Cryptosporidium,poliovirus
and Salmonella in marine waters. Wat. Sci. Tech. 35, 261–268.
La Pointe, B.F., O’Connell, J.D., Garrett, G.S., 1990. Nutrient
couplings between on-site waste disposal systems, groundwaters,
and nearshore surface waters of the Florida Keys. Biogeochemistry
10, 289–307.
Lipp, E.K., Kurz, R., Vincent, R., Rodriguez-Palacios, C., Farrah,
S.R., Rose, J.B., 2001a. The effects of seasonal variability and
weather on microbial fecal pollution and enteric pathogens in a
subtropical estuary. Estuaries 24, 266–276.
Lipp, E.K., Lukasik, J., Rose, J.B., 2001b. Human enteric viruses and
parasites in the marine environment. In: Paul, J.H. (Ed.), Methods
in Microbiol., vol. 30. Academic Press, London, pp. 559–588.
Lyons, M.M., Aas, P., Pakulski, J.D., Van Waasbergen, L., Miller,
R.V., Mitchell, D.L., Jeffrey, W.H., 1998. DNA damage induced
by ultraviolet radiation in coral reef microbial communities. Mar.
Biol. 130, 537–543.
McLaughlin, M.R., 2000. Evaluation of the Bacteriodes fragilis phage
assay as an alternative indicator of sewage pollution. M.S. Thesis,
Department of Marine Science, University of South Florida, p. 72.
Nobles, R.E., Brown, P., Rose, J.B., Lipp, E.K., 2000. The investi-
gation and analysis of swimming-associated illness using the fecal
indicator enterococcus in southern Florida’s marine waters. Flor-
ida J. Env. Health 169, 15–19.
Paul, J.H., DeFlaun, M., Jeffery, W.H., 1986. Elevated levels of
microbial activity in the coral surface microlayer. Mar. Ecol. Prog.
Ser. 33, 29–40.
Paul, J.H., Rose, J.B., Brown, J., Shinn, E., Miller, S., Farrah, S.,
1995. Viral tracer studies indicate contamination of marine waters
by sewage disposal practices in Key Largo, Florida. Appl. Env.
Microbiol. 61, 2230–2234.
Paul, J.H., Rose, J.B., Jiang, S., Zhou, X., Cochran, P., Kellogg, C.,
Kang, J.B., Griffin, D., Farrah, S., Lukasik, J., 1997. Evidence for
groundwater and surface marine water contamination by waste
disposal wells in the Florida Keys. Wat. Res. 31, 1448–1454.
Paul, J.H., McLaughlin, M.R., Griffin, D.W., Lipp, E.K., Stokes, R.,
Rose, J.B., 2000. Rapid movement of wastewater from onsite
disposal systems into surface waters in the Lower Florida Keys.
Estuaries 23, 662–668.
Porter, J.W., Lewis, S.K., Porter, K.G., 1999. The effects of multiple
stressors on the Florida Keys coral reef ecosystem: a landscape
hypothesis and a physiological test. Limn. Ocean 44, 941–949.
Rohwer, F., Breitbart, M., Jara, J., Azam, F., Knowlton, N., 2001.
Diversity of bacteria associated with the Caribbean coral Mon-
tastraea franksi. Coral Reefs 20, 85–91.
Shinn, E.A., Rees, R.S., Reich C.D., 1994. Fate and pathways of
injection-well effluent in the Florida Keys. USGS Report 94–276,
p. 105.
Solic, M., Krstuvolic, N., 1992. Separate and combined effects of solar
radiation, temperature, salinity and pH on the survival of fecal
coliforms in seawater. Mar. Poll. Bull. 24, 411–416.
US Environmental Protection Agency, 1994. Monitoring requirements
for public drinking water supplies: proposed rule. Federal Register
59 (28).
US Environmental Protection Agency, 1997. Method 1600: Membrane
Filter Test Method for Enterococci in Water. EPA-821-R-97-004.
670 E.K. Lipp et al. / Marine Pollution Bulletin 44 (2002) 666–670
... In contrast to Enterococci, Vibrio found abundantly in coral mucus as it is symbiont of corals and other marine organisms, some of them contribute nutrients to the host while other become the disease pathogens such as Porites Ulcerative White Spots (Arboleda & Reichardt 2010), and Porites White Patch Syndrome (Séré et al 2013). However our result was not in line with the studies of Lipp et al (2002) and Lipp & Griffin (2004) that found the concentration of Enterococci was higher in seawater relative to the coral mucus. Presumably, our results are related to the hydro-oceanography features of this area that reduced the resident time to Enterococci in the water column. ...
... Presumably, our results are related to the hydro-oceanography features of this area that reduced the resident time to Enterococci in the water column. However, both this studies Griffin 2004 andLipp et al 2002) did not provided any hydro-oceanographic data. It is suggested that the extremely abundant Vibrio in the coral mucus at all sites might be caused by organic carbon that produced in coral mucus and exacerbated with the input from anthropogenic activities. ...
Article
Full-text available
The increase of anthropogenic land-based activities in the Pemuteran village, a tourist spot in the northern part of Bali Island, threatened the health of its coral reefs. Therefore, this study aimed to assess the impact of anthropogenic activities in the coral reef by using two indicator bacteria (Enterococci and Vibrio). Study sites were chosen based on human activities whereas coral genus was selected following the three most abundant genera in each site. The mucus of coral fragments and the water column overlying the reefs were sampled. These samples were both tested for the presence of Vibrio in TCBS nutrient at 37°C for 24 hours and Enterococci in Slanetz and Bartley nutrient at 41°C for 24 hours. Results showed that the abundance of Enterococci in the mucus of all coral genera were relatively similar, whereas its abundances in seawater were significantly higher than those in mucus. In contrast to Vibrio, the abundances in mucus in all coral genera were significantly higher than those in seawater. Despite the likely relativeness to the natural characteristics of both indicator bacteria, the extremely high concentration of total organic carbon and nitrate in the water column, particularly nitrate, significantly enhance the abundance of Enterococci. The high level of nutrients was detected in the site which has the highest human activities, milkfish culture and marine recreational spot. The abundant of Vibrio may reveal the cause of the high prevalence of coral diseases in this area, whereas Enterococci indicate that anthropogenic pollution has reached its coral reef ecosystem.
... Some bacterial and viral pathogens transported in ballast water occur on the outer surfaces of invertebrates (Saccà 2015). For example, bacteria have been observed attached to the chitinous surfaces of copepods and other crustaceans (Huq et al. 2001;Lipp et al. 2002), to larval stages of planktonic invertebrates (Martinelli Filho et al. 2010), and to the mucilaginous envelopes of some phytoplankton (Huq et al. 2001;Lipp et al. 2002). Exposure to pathogens can result in fitness impacts to marine invertebrates with some life stages more susceptible than others. ...
... Some bacterial and viral pathogens transported in ballast water occur on the outer surfaces of invertebrates (Saccà 2015). For example, bacteria have been observed attached to the chitinous surfaces of copepods and other crustaceans (Huq et al. 2001;Lipp et al. 2002), to larval stages of planktonic invertebrates (Martinelli Filho et al. 2010), and to the mucilaginous envelopes of some phytoplankton (Huq et al. 2001;Lipp et al. 2002). Exposure to pathogens can result in fitness impacts to marine invertebrates with some life stages more susceptible than others. ...
Technical Report
Full-text available
Vessels involved in commercial marine shipping in Canada engage in the movement of goods or people by sea on the Arctic, Atlantic, and Pacific oceans. To explore the ways that the activities associated with commercial shipping can impact the marine environment, a suite of activity-based Pathways of Effects (PoE) conceptual models were developed. PoE conceptual models describe the pathways (linkages) between human activities, associated stressors, and their effects on endpoints, based on current knowledge. A visual representation of each PoE model is supported by text describing each pathway linkage based on scientific literature or expert opinion. Indigenous and local knowledge were not used in the current work. PoE models are useful tools for the scoping phase of a variety of environmental assessment, such as ecological risk assessment, environmental impact assessment, and cumulative effect assessments as they clearly outline activities and stressors and clarify connections between human activities and potential effects on ecological endpoints, and provide a science-based foundation for decision-making. The objective of these models and their supporting evidence is to provide a systematic review of the effects of shipping-associated activities on marine ecosystems. PoE models have been developed for five activities associated with commercial marine shipping in Canada: 1) anchoring and mooring, 2) vessel at rest, 3) grounding and sinking, 4) movement underway, and 5) discharge (divided into two PoE models: ‘debris’ and ‘other’). The PoEs were developed to be broad enough to be adapted for application in a range of environments and locations and detail the potential stressors and effects that could be considered in an assessment. The activity-based PoE models contain fourteen stressors (e.g., substrate disturbance, vessel strikes) and are related to three effects (change in fitness, mortality, and change in habitat) on ten generic endpoints (e.g., marine mammals, physical habitat). The models only include activities related to the commercial movement of goods and people by vessels, not included in this document are other vessel activities such as fishing, seismic surveying, dredging, port operations (e.g., when at-berth and while berthing). Non-commercial vessels (e.g., recreational vessels) are also not specifically included in these models. Though endpoints have been identified for illustrative purposes here, ultimately the assessor is responsible for comprehensively scoping the specific endpoints (e.g., valued components) and stressors to be considered in any assessment. PoE models do not include any evaluation of the relative or absolute impact from these activities on specific endpoints; this would occur in a subsequent assessment step, such as risk assessment.
... Finally, the Florida coast has had a long history of watershed alteration and pollution, with significant effects on corals (Rogers, 1990;Marubini and Davies, 1996;Lipp et al., 2002;Downs et al., 2005;Wagner et al., 2010). Associated factors, such as contaminants, pesticides, toxicants, nutrients, and sedimentation, may selectively kill or stress zooxanthellae (Owen et al., 2003;Reichelt-Brushett and McOrist, 2003;Brodie et al., 2012), but their role, if any, in coral disease has been less well studied. ...
Article
Full-text available
Samples from eight species of corals (Colpophyllia natans, Dendrogyra cylindrus, Diploria labyrinthiformis, Meandrina meandrites, Montastraea cavernosa, Orbicella faveolata, Pseudodiploria strigosa, and Siderastrea siderea) that exhibited gross clinical signs of acute, subacute, or chronic tissue loss attributed to stony coral tissue loss disease (SCTLD) were collected from the Florida Reef Tract during 2016-2018 and examined histopathologically. The hallmark microscopic lesion seen in all eight species was focal to multifocal lytic necrosis (LN) originating in the gastrodermis of the basal body wall (BBW) and extending to the calicodermis, with more advanced lesions involving the surface body wall. This was accompanied by other degenerative changes in host cells such as mucocyte hypertrophy, degradation and fragmentation of gastrodermal architecture, and disintegration of the mesoglea. Zooxanthellae manifested various changes including necrosis (cytoplasmic hypereosinophilia, pyknosis); peripheral nuclear chromatin condensation; cytoplasmic vacuolation accompanied by deformation, swelling, or atrophy; swollen accumulation bodies; prominent pyrenoids; and degraded chloroplasts. Polyhedral intracytoplasmic eosinophilic periodic acid-Schiff-positive crystalline inclusion bodies (∼1-10 µm in length) were seen only in M. cavernosa and P. strigosa BBW gastrodermis in or adjacent to active lesions and some unaffected areas (without surface lesions) of diseased colonies. Coccoidlike or coccobacilloidlike structures (Gram-neutral) reminiscent of microorganisms were occasionally associated with LN lesions or seen in apparently healthy tissue of diseased colonies along with various parasites and other bacteria all considered likely secondary colonizers. Of the 82 samples showing gross lesions of SCTLD, 71 (87%) were confirmed histologically to have LN. Collectively, pathology indicates that SCTLD is the result of a disruption of host-symbiont physiology with lesions originating in the BBW leading to detachment and sloughing of tissues from the skeleton. Future investigations could focus on identifying the cause and pathogenesis of this process.
... ng L −1 ), caffeine (5.5-68 ng L −1 ), Diethyl-m-toluamide (DEET) (4.8-49 ng L −1 ), and especially estrone (E1) (<0.1-5.2 ng L −1 ) due to its high detection frequency (81%) as compounds of concern, since concentrations found in surface waters were particularly high for common coastal environments (Singh et al., 2010). Likewise, other studies in the Florida Keys measuring EDCs through indirect techniques also suggested coral exposure to hormones released by human activities could be significant and possibly impact the health of native corals (Atkinson et al., 2003;Lipp et al., 2002). Moreover, there is a growing public concern on the effects of chronic water quality degradation of this sensitive coastal environment that needs to be addressed. ...
Article
Environmental exposure risk to different xenobiotics, which can potentially alter the function of the endocrine system, remains a great health and safety concern for aquatic species and humans. Steroid hormones, pharmaceuticals and personal care products (PPCPs) have been identified as important aquatic contaminants due to their widespread occurrence in surface waters and their endocrine disrupting properties. Heavily populated areas in South Florida not served by municipal wastewater collection present an unexpected high risk of anthropogenic contaminants to nearby coastal systems through surface runoff and groundwater flow. Previous studies in South Florida have been largely concentrated on assessing the relevance of the fate and transport of inorganic nutrients, heavy metals and pesticides with regulatory criteria. Therefore, a significant gap exists in assessing occurrence, distribution and biological significance of the presence of human related organic contaminants in natural surface waters. In this study, we have developed a fast and sensitive online solid-phase extraction followed by liquid chromatography- high resolution mass spectrometry (SPE-LC-HRMS) method using a Q-Exactive system for the determination of the occurrence and distribution of selected wastewater tracers/indicators, recalcitrant PPCPs and steroid hormones in South Florida surface waters. Seasonal and spatial variations of these contaminants were monitored from 2017 to 2019. The presence of total coliforms and E. coli were also evaluated in order to further assess water quality. Correlations between hormones and anthropogenic tracers were explored to better elucidate the sources, pathways and exposure risks to these contaminants. Caffeine, sucralose, Diethyl-m-toluamide (DEET) and carbamazepine were frequently detected in the water samples, which is indicative of extensive wastewater intrusion impacting the surface water. Estrone (E1), 17-β- estradiol (E2), and 17-α-ethynylestradiol (EE2) levels found in surface water raises concern of potential endocrine disruption effects in the aquatic ecosystem. Hazard quotient have been calculated to identify areas with high ecological risks to aquatic organisms.
... Though these standards are directed at human health safety, there is growing evidence that contaminated waters from sewage can carry pathogens that affect marine organisms (U.S. EPA 2003;2012a). For example in Florida, fecal contamination has been traced to coral reefs (Lipp et al. 2002;Lipp and Griffin 2004); Serratia marcescens found in coral mucus has been identified as a causative agent acropora serratiosis (originally White Pox) for disease in Acropora palmata (Patterson et al. 2002;Sutherland 2003;Sutherland et al. 2010Sutherland et al. , 2011. Though the risk-potential for sewage associated pathogens affecting coral health is unknown, the linkage found between sewage and coral disease in the Caribbean warrants closer inspection in other coral reef habitats. ...
Technical Report
Full-text available
West Maui, HI has been plagued with reports of poor water quality in the nearshore coastal zone, fecal indicators exceeding EPA standards, and algal blooms for over 20 years with a corresponding steady decline in coral covery from 70% (1990s) to 27% (2006). This final report provides baseline data related to bacterial water quality in wet and dry seasons and toxicity bioassay data at multiple locations along the Maui coastline from sediment and water sampled in 2012 and 2013. These data will help clarify the role of wastewater injection wells may play in coral decline and assist in best management practices for monitoring efforts. This information can help strategically focus costly management efforts on the greatest risk factors for mitigation and restoration of these vulnerable marine resources.
... Coral mucus also plays an important part in coral disease which has been responsible for significant coral mortality 30,31 . Coral mucus may function both as a protective physicochemical barrier 30,32,33 and as a growth medium for bacteria, including potential pathogens 34,35,36 . Even the most fundamental measure, the rate of mucus production, is extremely difficult to assess and is poorly defined in the literature. ...
Article
Full-text available
Surface water quality deterioration around the Kavaratti island was studied through the determination of the traditional indicators of faecal pollution, total coliforms (TC), faecal coliforms (FC) and Faecal streptococci (FS) along with supplemental indicator bacteria, Vibrio cholerae (VC), Vibrio parahaemolyticus (VP), Escherichia coli (EC) and Shigella species (SH). Cluster analyses were performed to differentiate the bacterial loading of different area. Helipad region indicates a separate cluster with the abundance of all bacteria (TC of 740CFU/mL, FC of 290CFU/mL, EC of 120CFU/mL, SH of 160CFU/mL, VC of 650CFU/mL, VP of 190CFU/mL and FS of 120CFU/mL). Significant positive relationship was observed between total nitrogen and indicator bacteria (R² > 0.844).
... Mucus are responsible for significant of coral mortality, mainly in the Caribbean [45]. Mucus layer may both act as a protective physicochemical barrier [8,46,47] and growth media for bacteria which include pathogens [23,48,49]. Particularly, the SML provides hundred times of growth media for bacteria than other layers and here able to act as metabolically in nearby seawater [50]. ...
Article
Full-text available
Coral reefs being highly diverse to marine ecology, yet most of the mystery remain unsolved. Coral reefs are of ecological as well as economic importance. Corals support commercial and subsistence fisheries, tourism, and are of medicinal value. It also creates a barrier against storms, flooding, and erosion by reducing wave action. Further, Corals support the existence of nearly 4,000 species of fish, 800 species of hard corals reef and other oceanic species. They also act as carbon sinkers. Coral polyps take up dissolved CO 2 in the water to make their calcareous exoskeleton allowing atmospheric CO 2 levels to be reduced. People around the world depend on the coral reefs as it's used as food, protection, provides employment and attracts tourism. They are attractive sites for recreational activities such as snorkeling and scuba diving. Furthermore, the coral reef ecosystem could represent increasingly significant sources for medical treatments, nutritional supplements, pesticides, cosmetics and other commercial products. Therefore, needs best strategies to improve to protect coral reef.
Article
Objective: It was aimed to evaluate the sea water and beach sand of beaches in Antalya in terms of microbiological. Methods: Sea water samples and sand samples from Konyaaltı and Lara beach were collected from the designated sampling points. Sand samples were taken from three different layers which were knee length bottom sand (DBK), wave zone (DZ) and dry sand (KK). These samples were examined by membrane filtration in terms of Esherichia coli, intestinal enterekok, Salmonella spp., yeast-Candida spp., Pseudomonas aeruginosa. Results: When the general average distribution of sand, seawater and sand layer samples taken from Konyaaltı and Lara beaches was examined, it was determined that the microbial pollution in sea water was high. Conclusion: In resulting of our work, E. coli, in the samples, were 14% in Konyaalti beach and 6% in Lara beach. Intestinal enterococcus were also 22% in Konyaaltı beach and 30% in Lara beach. ÖZET Amaç: Antalya kent plajlarının deniz suyu ve plaj kumunun mikrobiyolojik yönden değerlendirilmesi amaçlanmıştır. Yöntem: Konyaaltı ve Lara plajından deniz suyu örnekleri ile kum örnekleri, belirlenen örnekleme noktalarından toplandı. Kum örnekleri, diz boyu dip kumu (DBK), dalga zonu (DZ) ve kuru kum (KK) olmak üzere üç farklı tabakadan alındı. Alınan örnekler, membran filtrasyon yöntemiyle Esherichia coli, intestinal enterekok, Salmonella spp., maya-Candida spp. ve Pseudomonas aeruginosa açısından incelendi. Bulgular: Konyaaltı ve Lara plajlarından alınan kum, deniz suyu ve kum katman örneklerin genel ortalama dağılımı incelendiğinde, deniz suyunda mikrobiyal kirliliğin yüksek olduğu belirlendi. Sonuç: Çalışmamız sonucunda; alınan örneklerinde E. coli için sırasıyla Konyaaltında %14'ü, Lara'da %6'sında, intestinal enterokok ise Konyaaltı'nda %22, Lara'da %30 oranında bir kirlilik görüldü.
Article
Full-text available
Mass mortalities due to disease outbreaks have recently affected major taxa in the oceans. For closely monitored groups like corals and marine mammals, reports of the frequency of epidemics and the number of new diseases have increased recently. A dramatic global increase in the severity of coral bleaching in 1997–98 is coincident with high El Niño temperatures. Such climate-mediated, physiological stresses may compromise host resistance and increase frequency of opportunistic diseases. Where documented, new diseases typically have emerged through host or range shifts of known pathogens. Both climate and human activities may have also accelerated global transport of species, bringing together pathogens and previously unexposed host populations.
Article
Discharge of sewage into the ocean is still a common method of disposal worldwide. Both treated and untreated sewage may contain significant concentrations of waterborne pathogens, such as Giardia, Cryptosporidium, poliovirus and Salmonella. Limited studies exist on the survival of poliovirus and Salmonella in marine waters; however, almost no information exists on the survival of protozoan parasites in marine waters. This study examined the survival of Giardia muris cysts, Cryptosporidium parvum oocysts, poliovirus-1 and Salmonella typhimurium in marine waters. The survival of the microorganisms varied according to the presence of light, salinity and water quality (as determined by quantity of enterococci). All microorganisms survived longer in the dark than in sunlight, the order of survival in sunlight being: Cryptosporidium > poliovirus > Giardia > Salmonella.
Article
In order to assess the microbial water quality in canal waters throughout the Florida Keys, a survey was conducted to determine the concentration of microbial fecal indicators and the presence of human pathogenic microorganisms. A total of 19 sites, including 17 canal sites and 2 nearshore water sites, were assayed for total coliforms, fecal coliforms, Escherichia coli , Clostridium perfringens , enterococci, coliphages, F-specific (F ⁺ ) RNA coliphages, Giardia lamblia , Cryptosporidium parvum , and human enteric viruses (polioviruses, coxsackie A and B viruses, echoviruses, hepatitis A viruses, Norwalk viruses, and small round-structured viruses). Numbers of coliforms ranged from <1 to 1,410, E. coli organisms from <1 to 130, Clostridium spp. from <1 to 520, and enterococci from <1 to 800 CFU/100 ml of sample. Two sites were positive for coliphages, but no F ⁺ phages were identified. The sites were ranked according to microbial water quality and compared to various water quality standards and guidelines. Seventy-nine percent of the sites were positive for the presence of enteroviruses by reverse transcriptase PCR (polioviruses, coxsackie A and B viruses, and echoviruses). Sixty-three percent of the sites were positive for the presence of hepatitis A viruses. Ten percent of the sites were positive for the presence of Norwalk viruses. Ninety-five percent of the sites were positive for at least one of the virus groups. These results indicate that the canals and nearshore waters throughout the Florida Keys are being impacted by human fecal material carrying human enteric viruses through current wastewater treatment strategies such as septic tanks. Exposure to canal waters through recreation and work may be contributing to human health risks.
Article
Changes in land use and water management practices in south Florida have altered the quality and quantity of freshwater flowing into Florida Bay. By the 1980s, reduced inflow and drought led to an extensive hypersaline phase in the bay. This phase had a drastic effect on benthic communities within the bay and possibly also on coral communities within the bay and the Florida Keys National Marine Sanctuary. Physical oceanographic measurements demonstrate the presence of warm, hypersaline, and turbid water on coral reefs offshore from the Florida Keys, especially near passes which conduct water from Florida Bay to the Atlantic Ocean. To examine the effect of Florida Bay water intrusions on coral reefs, we tested for significant effects of two stressors, elevated temperature and salinity, on coral production, respiration, and survival. Elevated temperatures produce significant reductions in photosynthesis, respiration, and net P:R ratios afte r6ho fexposure, and elevated salinities produce similar results after 30 h. Exposure to both elevated temperature and salinity produces a highly significant (P. 0.01), but short-term, mitigative interactive effect. The combination of the two stressors was less stressful (for the response variables measured) than the sum of the stressors acting independently. After 36 h of exposure, however, the mitigating effect disappeared and corals exposed to the combined stresses did not survive. A three-dimensional response surface, which predicts P:R ratios as a function of varying salinity and temperature, is used to construct a testable hypothesis to explain recent declines in coral cover on some reefs within Florida Bay and the Florida Keys. We chose salinity and temperature to test a multiple stressor model because they are relatively easy to manipulate. However, any enviromentally realistic model must include other potential stressors, such as turbidity, elevated nutrients, and environmental contaminants.
Article
Water is a common vehicle for the transmission of many enteric viruses. Enteric viruses survive longer in fresh and marine water than coliform bacteria, which are used to monitor water quality. In marine and other surface waters, numbers of enteric viruses are often too low to be detected in unconcentrated samples. Therefore, large volumes of water must be concentrated by adsorption/filtration and elution (desorption) before analysis. More than 100 types of human pathogenic viruses are present in fecally contaminated water, and are detected by the current available methods. The detection of adenoviruses, hepatitis A virus, astroviruses, and rotaviruses by cell culture is possible; however, the methods are tedious and less often used. Nucleic acid hybridization and the polymerase chain reaction (PCR) are used for detecting human viruses and other fastidious microbial pathogens in the environment. Molecular detection methods are important in studying the occurrence of Hepatitis A and Norwalk-like viruses, which are epidemiologically important pathogens but do not produce cytopathogenic effects (CPE) readily in cell culture. Molecular techniques have also enhanced the speed and sensitivity of detection for the more routinely cultured enteroviruses.
Article
Traditional fecal coliform bacterial indicators have been found to be severely limited in determining the significance and sources of fecal contamination in ambient waters of tropical and subtropical regions. The bacteriophages that infectBacteroides fragilis have been suggested as better fecal indicators and at least one type may be human specific. In this study, the phages that infectB. fragilis host RYC2056 (RYC), including phage B56-3, and host ATCC 51477-HSP40 (HSP), including the human specific phage B40-8, were evaluated in the drainage basins of Tampa Bay, 7 samples (n=62), or 11%, tested positive for the presence of phages infecting the host HSP, whereas 28 samples, or 45%, tested positive using the host RYC. A survival study was also done to compare the persistence of phages B56-3 and B40-8 to MS2 coliphage in seawater at various temperatures. The decay rates for MS2 were 0.239 log10 d−1 at 10°C, but increased to 0.896 at 20°C and 2.62 log10 d−1 at 30°C. The twoB. fragilis phages persisted much longer in the seawater compared to the coliphage and showed little variation between the temperatures. All sewage influents sampled from area wastewater treatment plants contained phages that infected the twoB. fragilis hosts at levels from 1.2×104 to 1.11×105 pfu 100 ml−1 for host RYC and 67 to 350 pfu 100 ml−1 for host HSP. Of the 7 chlorinated effluent samples tested, 3 were positive for the presence of the phage using the host RYC and the phage enrichment method, with levels estimated to be −1. No phages were detected using the host HSP in the treated sewage effluent. Coliphages were found in 3 of the 7 effluent samples at a range of 30 to 1.2×103 pfu 100 ml−1.
Article
The external mucus layers of the stony coral Porites astreoides and the soft corals Palythoa sp. and Heteroxenia fuscesens are inhabited by communities of marine heterotrophic bacteria. Population levels of bacteria in coral mucus may be regulated by the self-cleaning behavior of the host. Bacterial populations in coral mucus respond to stresses applied to the host coral by growing to higher population levels in the mucus, indicating that these are populations of viable organisms closely attuned to host metabolism. Members of these microbial populations utilize the mucus compounds and may play a role in processing coral mucus for reef detritus feeders. One such species, Vibrio alginolyticus, grows rapidly on Heteroxenia mucus, is attracted to dissolved mucus, and possesses a mechanism to maintain itself on the coral surface.