Environ Monit Assess (2022) 194: 71

https://doi.org/10.1007/s10661-021-09679-7

Seasonal variations in the water quality and antibiotic

resistance of microbial pollution indicators in the Mandovi
and Zuari estuaries, Goa, India
Ashwini D. Toraskar · Cathrine S. Manohar ·
Charmaine L. Fernandes · Durbar Ray ·
Abigayle D. Gomes · Ajit Antony

Received: 2 June 2021 / Accepted: 6 December 2021 / Published online: 7 January 2022
© The Author(s), under exclusive licence to Springer Nature Switzerland AG 2022

Abstract The two adjacent estuaries of the rivers during the post-monsoon season. However, the esti-
Mandovi and Zuari, along the Goa coast in the cen- mated multiple antibiotic resistance (MAR) index
tral west coast of India, are a large complex aquatic suggests that bacterial isolates in monsoonal water
system hosting diverse natural habitats. The water and sediment samples have maximum resistance
quality in these habitats is affected by various anthro- towards antibiotics. This shows that, though the basic
pogenic activities as they are extensively used for water quality improved during the monsoon, possibly
transportation, fisheries and various recreational due to substantial dilution, the increased terrestrial
activities. In the present study, changes in the water inputs brought harmful pathogens into these estuarine
quality and levels of microbial pollution during the waters, which may be of potential health risk. Under-
pre-monsoon, monsoon and post-monsoon seasons standing the ecological status of the estuarine habitats
were determined. The water quality index was esti- is important for successful environmental manage-
mated based on the parameters: temperature, salinity, ment and sustainable development.
pH, dissolved oxygen, biochemical oxygen demand
and nutrients. The seasonal changes in the micro- Keywords Mandovi · Zuari · Estuary · Water
bial pollution load were also assessed based on the quality index · Microbial pollution indicators ·
abundance of pollution indicator organisms and their Antibiotic resistance
resistivity towards multiple antibiotics. Results show
that the water quality index status was ‘poor’ in the
pre-monsoon and post-monsoon seasons and it was Introduction
‘good’ only in the monsoon period. Levels of pollu-
tion indicator organisms determined show that the Estuaries represent productive aquatic habitats and
counts were the highest in the pre-monsoon season, are characterised by dynamic biogeochemical pro-
which reduced in the monsoon and further declined cesses. They provide some of the most invaluable
ecosystem resources and services. Besides that, estu-
aries have drawn much attention as they support the
Supplementary Information The online version
contains supplementary material available at https://​doi.​
livelihood of a large majority of the human popula-
org/​10.​1007/​s10661-​021-​09679-7. tion globally and hence are constantly exposed to
anthropogenic perturbations (Liu et al., 2016). Such
A. D. Toraskar · C. S. Manohar · C. L. Fernandes · semi-enclosed environments with limited flushing
D. Ray (*) · A. D. Gomes · A. Antony
capacity are influenced by seasonal variations, espe-
Dona Paula, Goa 403004, India
e-mail: dray@nio.org cially due to freshwater influx in the monsoon season

Vol.: (0123456789)
13
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
 71 Page 2 of 17 Environ Monit Assess (2022) 194: 71

and tend to accumulate a variety of anthropogenic Streptococcus sp. (Nagvenkar & Ramaiah, 2009;
wastes (VishnuRadhan et al., 2015). The 7500-km Rodrigues et al., 2011). Adding to this, regular urban
long Indian coastline is drained out by major rivers: effluents bringing in medical wastes, especially anti-
about 14 of them, 44 medium and nearly 162 minor biotic loads, increase the threat to the ecosystem
rivers and thus characterised with many complex (Vignesh et al., 2014). This not only introduces anti-
estuarine habitats (Kumar & Sarma, 2018). Various biotic-resistant microbial strains into the environment
factors such as the geophysical locations, riverine but also accelerates the spread of resistance genes
fluxes and other man-made developments influence to indigenous bacterial flora. As a consequence, the
the environmental status of Indian estuaries. The emergence of multidrug resistance is a major threat
estuaries along the west coast of India are largely faced by today’s world (De Souza et al., 2006). Con-
influenced by intense monsoonal precipitation during sidering the current scenario, regular monitoring of
the southwest monsoon season and hence experience the estuarine habitats to determine the health of the
significant seasonal variations in the physicochemical coastal water bodies is required. It can directly impact
parameters (Shetye et al., 2007). the surrounding riverine systems, and its deteriora-
Along the central west coast of India, the Mandovi tion can have a major threat to the human population
and Zuari estuaries of Goa are two important estua- dependent on them. In this study, we have estimated
rine habitats (Kessarkar et al., 2010). They receive the impact of monsoon season on the water quality
maximum freshwater runoff (up to ~ 400 ­m3 ­s−1 dis- index (WQI) and the multiple antibiotic resistance
charge) in June–September each year during the index (MAR) of microbial pollution indicators in the
active phase of the monsoon (Manoj et al., 2009), and estuarine environments.
with the withdrawal of monsoon, the flow becomes
primarily tidal, until the following monsoon season
(Sundar & Shetye, 2005). Such seasonal variability Materials and methods
along with regular tidal influence brings about large
variations in the physicochemical and microbiologi- Study location and sampling details
cal parameters of the two estuaries. Due to incessant
rains during the monsoon season, the water column in Sampling for this study was carried out within the estu-
these estuaries is stratified; however, the waters stay arine regions of the Mandovi and Zuari rivers along the
well mixed in the summer before the monsoonal rains Goa coast (Fig. 1). These two adjacent tropical estuar-
and also throughout the post-monsoonal season. The ies experience strong seasonal changes: very warm and
intrusion of saltwater up to the far upstream region humid pre-monsoon (February–May), intensive precip-
makes these estuaries a marine-dominated estuarine itation during monsoon (June–October) and relatively
complex (Shetye et al., 2007; VishnuRadhan et al., dry, post-monsoon (November–January) conditions.
2015). These semi-enclosed coastal water bodies, To understand the possible impacts of these seasonal
which are connected to the open sea and the seawater variations on the water quality and microbial commu-
in these habitats, are measurably diluted by freshwa- nity structure, sampling was carried out in April 2019
ter due to terrestrial drainage. This freshwater influx (pre-monsoon), September 2019 (monsoon) and Janu-
from rivers helps in maintaining the ecological char- ary 2020 (post-monsoon season). The water and sedi-
acteristics of the estuary. ment samples were collected from 16 sampling loca-
Earlier studies show that due to the rapid increase tions around this estuarine system (Fig. 1) on a trawl
in anthropogenic activities, the level of terrigenous boat. At each sampling location, water samples from
material entering these estuarine environments along the surface (~ 0.5 m water depth) and close to the bot-
the river discharge has been rising at an alarming rate tom (~ 0.5 m above the sediment) using pre-cleaned
in the Mandovi and Zuari estuaries which are con- Niskin samplers (capacity: 5 L) were collected. Sedi-
sidered to be the lifeline of Goa (Alagarsamy, 2006; ment samples were collected using a steel-plated Van
Shynu et al., 2015). Studies also show deterioration Veen grab (capacity: 0.045 sq mt) after water sampling
of environmental quality due to an increase in the at each location. Water and sediment samples were
levels of various pathogenic microbes such as coli- immediately subsampled for physicochemical and
forms, Vibrio sp., Salmonella sp., Shigella sp. and microbiological analysis with necessary precautions

Vol:. (1234567890)
13
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
Environ Monit Assess (2022) 194: 71 Page 3 of 17 71

Fig. 1  Sampling locations in the Mandovi and Zuari estuaries of Goa, India

and immediately processed or preserved at 4℃ follow- Similarly, the BOD was estimated with 5 days incu-
ing the standard protocols. bation at 27℃. The essential nutrients, including dis-
solved nitrate, nitrite, phosphate and silicate, were
analysed in all the water samples following the photo-
Analysis of physicochemical parameters in water
metric methods described by Grasshoff (1983).
samples

To assess the water quality in this estuarine system,

Estimation of WQI
the parameters such as temperature, salinity, pH, dis-
solved oxygen (DO), biochemical oxygen demand
The seasonal changes in the WQI of Mandovi and
(BOD) and essential nutrients were analysed in the
Zuari estuaries were calculated based on the physico-
water samples from all the sampling stations during
chemical parameters. The water quality parameters
each season. During field sampling, the temperature,
determined included temperature, salinity, pH, DO,
salinity and depth of the water column at each sam-
BOD, nitrate and phosphate concentration by using
pling station were determined using a portable CTD
the following equation (Adamou et al., 2020):
system (Sea Bird Electronics). The pH was measured
immediately after returning to the laboratory using a ∑
qW
benchtop pH metre (Thermo Fisher Scientific) with WQI = ∑ n n
Wn
an accuracy of ± 0.1 pH unit. Winkler’s titrimetric
method was used to determine the DO levels in the where the quality rating of the nth parameter, qn, was
water samples, and the values are expressed in mg/l. calculated for each parameter as

Vol.: (0123456789)
13
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
 71 Page 4 of 17 Environ Monit Assess (2022) 194: 71

Table 1  List of water quality parameters and their permissible Microbiological analysis
limits
Parameters Standard values Reference The microbiological load in the Mandovi and Zuari
estuaries was estimated based on the total viable
DO (mg/L) >5 USEPA (1986)
counts (TVC) of bacteria. The microbial pollution in
BOD (mg/L) <3 USEPA (1986)
both the estuaries was determined based on the enu-
Temp (℃) 30 Dheenan et al. (2014)
meration of pollution indicator organisms (PIOs).
pH 6.5–8.5 USEPA (1986)
The multiple antibiotic resistance (MAR) index of
Salinity (PSU) 35 Dheenan et al. (2014)
the PIOs was also estimated. Estimates of bacte-
Nitrate (µM) <2 Jha et al. (2015)
rial TVC in water and sediment samples were car-
Phosphate (µM) < 1.2 USEPA (1986)
ried out using standard microbiological, spread plat-
ing method on Zobell Marine Agar (HiMedia). The
colony-forming units (CFUs) were enumerated after
Vn − Vid 48 h of incubation at 28 ± 0.5℃ and are represented
qn × 100 as CFU/ml for water and CFU/g dry weight for sedi-
Sn − Vid
ment samples. The PIOs from the same samples were
In the above equation, Vn and Vid are the esti- isolated on selective bacteriological media (HiMedia,
mated and ideal values of the nth parameter, respec- India) according to the manufacturer’s guide. The dif-
tively; Vid value of 0 was used for all the parame- ferent groups of PIOs such as total coliforms, faecal
ters; however, for pH the value 7 and for dissolved coliforms/Escherichia coli like organisms (ECLO)
oxygen the value 12 mg/l, which is the maximum and other major PIOs: Streptococcus faecalis (SFLO),
saturation limit of dissolved oxygen in seawater at Staphylococcus aureus (SALO), Pseudomonas aer-
0℃ and 25 PSU (Fondriest Environmental, 2013), uginosa (PALO), Salmonella sp. (SLO), Shigella sp.
were used. Sn is the standard permissible value of (SHLO), Klebsiella aerogenes (KALO), Proteus vul-
nth water quality parameter, which was followed for garis (PVLO) and Vibrio cholerae (VCLO); Vibrio
each parameter based on the USEPA, and national parahaemolyticus (VPLO) and Vibrio vulnificus
standards recommended for water quality (Table 1). (VVLO) represented together as Vibrio-like organ-
Wn represents the relative weight of the nth param- isms (VLO) were determined based on the colony
eter, computed as. morphology on specific media. The number of PIOs
Wn = k/Sn. where k is the proportionality con- from the samples was estimated by the spread plate
stant calculated from Sn: method on selective media, and CFUs with specific
colony characteristics were enumerated. Samples
1
∑ 1 were diluted in saline to obtain isolated colonies, and
Sn=1,2…n the CFUs were counted after 24–48 h of incubation
at 37 ± 0.5℃ for all the PIOs, except thermotolerant
The WQI values obtained were used to deter- ECLO which was grown at 44℃. Statistical analysis
mine the quality in both the estuaries across the to determine the significance in the seasonal varia-
three seasons following the standard ranking scale tions of physicochemical and microbiological param-
(Table 2). eters was carried out based on one factor ANOVA
using Microsoft Office Excel.

Table 2  Water quality index ranking criteria Evaluation of antibiotic resistivity in PIOs: estimation
WQI range Category of MAR index
< 50 Excellent
Representative bacterial morphotypes of the major
50–100 Good
PIO groups from all three seasons were studied for
100–200 Poor
their antibiotic susceptibility profiles. Altogether,
200–300 Very poor
93 morphotypes were tested against a broad spec-
> 300 Unfit for usage
trum of 16 antibiotics belonging to different classes

Vol:. (1234567890)
13
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
Environ Monit Assess (2022) 194: 71 Page 5 of 17 71

of bacteriostatic and bactericidal agents, at recom- The seasonal monitoring of the Mandovi and Zuari
mended concentrations, procured from HiMedia, estuarine system based on water column parameters
India. The antibiotics used to determine the MAR at 16 sampling stations showed that the average water
index included bacteriostatic agents such as Chlo- temperature and salinity ranged between 28–31℃
ramphenicol, Tetracycline, Azithromycin and Co- and 24–36 PSU, respectively. The average water tem-
trimoxazole and bactericidal agents like Ciprofloxa- perature and salinity show distinct seasonal changes
cin, Nalidixic acid, Ofloxacin, Clindamycin, Netillin, in both the estuaries; however, the pH was within a
Rifampicin, Vancomycin, Ampicillin, Ceftizoxime, narrow range of 8 ± 0.5 in all three sampling seasons
Ceftriaxone, Piperacillin and Ertapenem. The disk (Fig. 2). Temperature is a principal component affect-
diffusion method on Mueller Hinton Agar plates ing the solubility of oxygen and the rates of chemi-
was used for antibiotic susceptibility testing (Bauer, cal and biological processes. The seasonal changes of
1966). For this, initially, 200 µl of pure bacterial sus- temperature in the surface and bottom waters of Man-
pensions of each morphotype was grown to reach dovi and Zuari estuarine system measured showed
an optical density of 0.5 McFarland standard, corre- that the surface waters in the pre-monsoon and mon-
sponding to ­108 CFU/ml. Each bacterial suspension soon seasons were approximately 1℃ higher than
was spread plated using a sterile cotton swab onto the the bottom waters. However, in the post-monsoon
media to develop homogenous growth. Then, the anti- season, the surface and bottom waters were uniform
biotic discs were placed on the agar surface asepti- and the temperature was 28 ± 1℃ in the water column
cally. Plates were incubated at 37 ± 0.5℃ for 24 h, and (Fig. 2). Studies have recommended that the stand-
at the end of the incubation period, zones of bacterial ard temperature should not exceed 30℃ in the coastal
growth inhibition formed surrounding the antibiotic waters (Gupta et al., 2003). The higher temperature
discs were measured. Necessary controls were also values in the surface waters of pre-monsoon and
maintained as per the manufacturer’s instructions. monsoon seasons could be due to the high intensity
To evaluate the relative susceptibility, the multiple of solar radiation combined with sluggish mixing of
antibiotic resistance (MAR) index of each isolate waters (Qasim & Gupta, 1981). Similar patterns were
was estimated based on the total number of antibiot- seen in both Mandovi and Zuari estuaries. The tem-
ics tested and the number of antibiotics against which perature in the water column was within the permis-
it exhibits resistance (Chitanand et al., 2010; Maloo sible range of 30℃ in both the estuaries in all three
et al., 2017). seasons, except in Mandovi and Zuari surface waters
in the pre-monsoon season.
Salinity is the defining parameter of the estuarine
Results and discussion habitats, and it shows wide variations with lower val-
ues during monsoon season due to the river runoff and
Seasonal variations of physicochemical parameters in increased values during the pre-monsoon or summer
the estuarine waters season due to reduced inflow from rivers. Our obser-
vations in the Mandovi and Zuari estuaries during the
Estuarine environments are governed by complex pre-monsoon show that the surface water values were
physicochemical and biological processes. Hydro- 33.6 PSU and 34.2 PSU, respectively, and the bottom
graphical parameters, temperature, salinity, pH, DO, waters were close to 35 PSU (Fig. 2). In the monsoon
BOD and nutrient levels, can largely influence the season, the lowest salinity values of 25 ± 2 PSU were
water quality (Moss et al., 2006). Hence, to evalu- recorded in the surface waters and the bottom waters
ate the environmental conditions in the Mandovi and were 30 ± 2 PSU. Distinct stratification with a salt
Zuari estuaries, these physicochemical parameters wedge in the bottom layer was observed only during
were analysed in the surface and bottom water sam- the monsoon season. However, in the post-monsoon
ples at all 16 sampling locations in all three sampling season, the waters were well mixed and the water col-
seasons. The results obtained for each parameter ana- umn salinity was close to 34 PSU. Though both the
lysed are presented as the mean of eight individual estuaries have salinity values close to seawater, with
sampling locations from each of the estuaries (Figs. 2 the onset of the monsoon, a sharp decrease in salin-
and 3). ity with distinct stratification in the water column is

Vol.: (0123456789)
13
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
 71 Page 6 of 17 Environ Monit Assess (2022) 194: 71

(a) Mandovi (b) Zuari

Temperature (℃)

33 33
31 31
29 29
27 27
25 25
Pre-monsoon Monsoon Post-Monsoon Pre-monsoon Monsoon Post-Monsoon
Salinity (PSU)

40 40

20 20

0 0
Pre-monsoon Monsoon Post-Monsoon Pre-monsoon Monsoon Post-Monsoon
9 9
8 8
pH

7 7
6 6
Pre-monsoon Monsoon Post-Monsoon Pre-monsoon Monsoon Post-Monsoon
10 10
8
DO (mg/L)

8
6 6
4 4
2 2
0 0
Pre-monsoon Monsoon Post-Monsoon Pre-monsoon Monsoon Post-Monsoon
8 8
BOD (mg/L)

6 6
4 4
2 2
0 0
Pre-monsoon Monsoon Post-Monsoon Pre-monsoon Monsoon Post-Monsoon

Fig. 2  Seasonal variations of temperature, salinity, pH, dis- ples of (a) Mandovi and (b) Zuari estuaries; dotted lines indi-
solved oxygen (DO) and biochemical oxygen demand (BOD) cate permissible limits, and error bars represent standard devi-
in the surface (white bars) and bottom (grey bars) water sam- ation (n = 8)

seen due to the heavy freshwater runoff during the 6.5 and 8.5. The pH values measured were within a
southwest monsoon (Vijith et al., 2009). The warm narrow range of 8 ± 0.5 in all the three sampling sea-
and lower saline, relatively less dense surface waters sons in the Mandovi and Zuari estuaries (Fig. 2).
float over the dense bottom waters, thus restricting the Dissolved oxygen is the prime parameter that
vertical mixing within the water column (Fig. 2). Due determines the water quality in any aquatic habitat.
to the strong buffering capacity of seawater, the pH A DO level of > 5 mg/L is considered the optimum
range in the estuarine habitats is generally between value for a healthy water body (Alam et al., 2007;

Vol:. (1234567890)
13
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
Environ Monit Assess (2022) 194: 71 Page 7 of 17 71

(a) Mandovi (b) Zuari

8 8
6 6
Nitrate (µM)

4 4
2 2
0 0
Pre-monsoon Monsoon Post-Monsoon Pre-monsoon Monsoon Post-Monsoon

3 3
Nitrite (µM)

2 2

1 1

0 0
Pre-monsoon Monsoon Post-Monsoon Pre-monsoon Monsoon Post-Monsoon
2 2
Phosphate (µM)

1 1

0 0
Pre-monsoon Monsoon Post-Monsoon Pre-monsoon Monsoon Post-Monsoon
30 30
Silicate (µM)

20 20

10 10

0 0
Pre-monsoon Monsoon Post-Monsoon Pre-monsoon Monsoon Post-Monsoon

Fig. 3  Seasonal variation of nitrate, nitrite, phosphate and silicate in the surface (white bars) and bottom (grey bars) water samples
of (a) Mandovi and (b) Zuari estuaries; dotted lines indicate permissible limits, and error bars represent standard deviation (n = 8)

Avannavar & Shrihari, 2008; Şener et al., 2017). Val- attributed to water turbulence during increased run-
ues lower than this are indicative of waste discharges off. According to Lin et al. (2006), temperature also
high in organic load and nutrients in the sampling has a direct effect on DO levels, and accordingly, in
region with increased microbial degradation. The DO the pre-monsoon season, when the temperature was
levels in both Mandovi and Zuari estuaries during all high, DO levels were also low in the estuaries.
three seasons were above the recommended limits, The amount of oxygen required to biochemically
except in the pre-monsoon season in Zuari estuary, oxidise organic material present in water is quanti-
where the average DO value was 4.8 mg/L (Fig. 2). fied as biochemical oxygen demand (BOD), and it is
In both estuaries, relatively higher DO values were directly dependent on the existing DO levels (Hocking,
found at the surface than in the bottom waters. Such 2005). According to USEPA (1986) recommendations,
differences were more prominent during the monsoon the BOD levels should be < 3 mg/L (Table 1). In this
and pre-monsoon (Fig. 2). The DO levels greatly study, BOD values were > 3 mg/L in both the estuaries
improved in the monsoon season, and this could be before and after the rains in the pre- and post-monsoon

Vol.: (0123456789)
13
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
 71 Page 8 of 17 Environ Monit Assess (2022) 194: 71

seasons (Fig. 2). However, during the rains, when and anoxia from eutrophication and an increase in algal
the DO levels were good, BOD values reduced growth (Chakraborty, 2017; Şener et al., 2017). Total
to < 3 mg/L. Values less than 3 mg/L show that during nitrogen comprising nitrate, nitrite and ammonia and
the monsoon season, there was increased river runoff total phosphates are the key elements controlling pri-
which ventilated the water. The seasonal variations in mary production in the coastal habitats. Studies report
the different physical parameters were supported with that if the total nitrogen levels are ~ 300–500 µg/L and
very good statistical significance for temperature, DO the total phosphate level is around 20 µg/L, then the
and BOD (Table 3). water body can be considered eutrophic (Yang et al.,
Coastal ecosystems, especially the estuaries, receive 2008).
inputs through riverine runoff from the terrestrial Eutrophication in estuaries can occur due to
regions and are rich in organic matter and nutrients increased nutrient concentration and lead to harm-
(Ram et al., 2003). These runoffs from the terrestrial ful consequences which can destabilise the delicate
regions influence the concentration of major nutrients ecosystem processes. The water quality guidelines
like nitrates, phosphates and silica in the estuarine sys- for coastal habitats recommend that the total nitrate
tem. These help in enhancing the numerous biologi- and nitrite levels should be < 2 µM, respectively (Jha
cal activities in the ecosystem, like the re-suspension et al., 2015). The water samples analysed during this
of benthic diatom propagules in response to nutrient study show that the average nitrate levels exceeded
availability, as reported by Khandeparker et al. (2015). the limit in both the estuaries in all three seasons.
However, an excessive supply of nutrients can also Values were ~ 4 µM during the summer, before the
have a negative impact in many instances. Increased rains, ~ 3 µM during the monsoonal rains, which
nutrients can lead to deleterious effects such as hypoxia was the lowest, and ~ 5.5 µM in the post-monsoon

Table 3  ANOVA: Variables MANDOVI ZUARI

one factor to show the
significance of seasonal Df F value F critical p value Df F value F critical p value
changes between different
parameters in the Mandovi Temperature 2 23.03 3.20 < 0.001*** 2 1.12 3.20 0.33
and Zuari estuaries Salinity 2 2.67 3.20 0.08 2 0.20 3.20 0.82
pH 2 2.46 3.20 0.10 2 0.31 3.20 0.74
DO 2 0.50 3.20 0.61 2 9.97 3.20 < 0.001***
BOD 2 0.96 3.20 0.39 2 5.84 3.20 < 0.005**
Nitrite 2 5.25 3.20 < 0.008** 2 10.00 3.20 < 0.001***
Nitrate 2 1.69 3.20 0.20 2 13.34 3.20 < 0.001***
Phosphate 2 13.34 3.20 < 0.001*** 2 23.72 3.20 < 0.001***
DF degree of freedom, Silicate 2 5.62 3.20 0.006** 2 6.48 3.20 0.003**
TVC total viable counts, TVC-water 2 3.91 3.20 0.02* 2 5.87 3.20 0.005**
TC total coliforms,
TVC-sediment 2 0.96 3.47 0.40 2 2.64 3.47 0.09
SFLO Streptococcus
faecalis-like organisms, TC-water 2 3.71 3.20 0.03* 2 2.84 3.20 0.06
SALO Staphylococcus TC-sediment 2 1.35 3.47 0.28 2 0.98 3.47 0.39
aureus-like organisms, SFLO-water 2 2.95 3.20 0.06 2 0.77 3.20 0.47
PALO Pseudomonas
SFLO-sediment 2 2.94 3.47 0.07 2 1.44 3.47 0.26
aeruginosa-like organisms,
SHLO Shigella-like SALO-water 2 1.97 3.20 0.15 2 5.61 3.20 0.006**
organisms, and VLO Vibrio- SALO-sediment 2 2.44 3.47 0.11 2 1.04 3.47 0.37
like organisms PALO- water 2 1.58 3.20 0.22 2 4.11 3.20 0.02*
F value greater than F PALO-sediment 2 1.01 3.47 0.38 2 7.25 3.47 0.004**
critical value indicates
SHLO-water 2 2.59 3.20 0.08 2 4.90 3.20 0.01*
statistical significance
* SHLO-sediment 2 18.36 3.47 < 0.001*** 2 1.45 3.47 0.26
Significant at 5%
** VLO-water 2 6.06 3.20 0.004** 2 15.82 3.20 < 0.001***
Significant at 1%
*** VLO-sediment 2 3.23 3.47 0.05 2 5.60 3.47 0.01*
Significant at 0.1%

Vol:. (1234567890)
13
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
Environ Monit Assess (2022) 194: 71 Page 9 of 17 71

season (Fig. 3). The nutrient levels dropped signifi- Nutrients like silica are known to significantly
cantly in the monsoon season which is evident that influence the diversity and distribution of phyto-
it was due to the dilution resulting from the incom- plankton, and some dominant species are also known
ing water flow. The increased nitrate levels could be to alter the community structure. The highest level
attributed to improper agricultural practices such as of silica was recorded in the pre-monsoon season
increased usage of inorganic fertilisers and manure which was ~ 13 ± 4 µM and the level declined in the
which leaches into the natural habitats (Bhavya et al., monsoon season to ~ 10 ± 6 µM. The values in the
2016). The nitrite levels, on the other hand, showed post-monsoon season were ~ 8 ± 4 µM in both the
relatively lower concentrations than nitrates (Fig. 3). estuaries. There are no standard limits for silicate
All the samples analysed were within the recom- concentrations; however, the levels of nitrates and
mended standard limits. The values across the sea- phosphates, together with increased silicate levels, are
sons were almost negligible to 1.2 µM concentration. very crucial for determining the water quality of the
The absence of nitrite levels shows that the ecosys- estuarine habitat (Carey & Fulweiler, 2014). Levels of
tem was not anaerobic as nitrite is one of the major the chemical parameters determined for the Mandovi
indicators of denitrification activity (Jayakumar et al., and Zuari estuaries show that the quality of the waters
2004). The levels of total nitrate and nitrite in the is close to eutrophicated waters in the pre-monsoon
Mandovi and Zuari region was close to 4 µM, espe- season with increased nitrate and phosphate levels.
cially in the pre-monsoon and post-monsoon sea- Seasonal variations of the different chemical param-
sons, which shows that the ecosystem is moderately eters between the three seasons were supported with
eutrophic. The monsoonal flushing seems to be play- very good significance in both the Mandovi and Zuari
ing an important role in reducing the levels of nitrates estuaries based on ANOVA analysis (Table 3). The
and preventing the system from becoming eutrophic. runoff and increased flushing cause a reduction in
Phosphates are found in extremely low concentra- the nutrient load during the monsoon season, thereby
tions in waters, as it is actively taken up by the pri- impacting the water quality of this monsoonal estu-
mary producers. The maximum permissible limit for ary. Earlier studies have also indicated that the mixing
total phosphates in coastal environments is deter- of wastewaters draining out from local urban areas is
mined as 1.2 µM (USEPA, 1986). However, studies the major source of excess nutrients in these estuarine
report that the ecosystem can become eutrophic even waters (Maya et al., 2011).
at ~ 0.2 µM concentration (Yang et al., 2008). The
phosphate levels in the Mandovi and Zuari waters Seasonal variation of estuarine water quality:
were within the permissible limit values (Fig. 3). The evaluation of WQI
highest concentrations of 0.78 ± 0.25 µM phosphate
were observed during the pre-monsoon season in To evaluate the composite impact of the physico-
both estuaries. Levels declined in monsoon season to chemical parameters on the estuarine waters, the
0.29 ± 0.14 µmol/L and again increased in the post- water quality index (WQI) was estimated for the
monsoon season up to 0.56 ± 0.25 µM. Residual deter- Mandovi and Zuari estuaries during the three sam-
gents in the domestic wastewaters, industrial effluents pling seasons. The WQI is used to evaluate the
and fertiliser runoff are the major contributors of changes in water quality of the habitats based on the
phosphates. Furthermore, all estuaries are known to levels of these parameters and their combined effect
have phosphate load due to constant adsorption and (Kannel et al., 2007). A total of seven parameters,
dissolution at the sediment–water interface, and even temperature, salinity, pH, DO, BOD, nitrate and phos-
a small increase in the concentration can impair the phate, were considered to evaluate the WQI. Each of
water quality level of the habitat, making the system these parameters plays a crucial role in determining
eutrophic Also, due to continuous adsorption and dis- ecosystem health, and permissible limits are provided
solution at the sediment–water interface, almost all by international and national environmental agencies
estuaries are known to have phosphate load and even (Table 1).
a slight increase in the concentration can reduce the The WQI was evaluated by comparing the average
water quality level of the habitat, making the system values of each of the parameters with the standard
eutrophic (Manoj et al., 2009; Sharpley et al., 2001). limits. A cumulative index was estimated based on

Vol.: (0123456789)
13
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
 71 Page 10 of 17 Environ Monit Assess (2022) 194: 71

the deviation of these parameters from the set limits The WQI improved during the monsoon period;
which was used to derive the WQI for the study area however, in the post-monsoon season that followed,
across the three seasons (Adamou et al., 2020). WQI water quality status deteriorated in both the estuar-
can range from 0 to 300, and the habitat studied is ies. The WQI during the post-monsoon season was
assessed into different categories such as excellent, categorised as ‘poor’ as the values were 130 and 120
good, poor, very poor and unfit for usage (Table 2). in the Mandovi and Zuari estuaries, respectively.
The quality of the water in aquatic ecosystems is This decline can be linked to the leaching of domes-
considered to be excellent if the WQI was less than tic waste discharge and agricultural fertilisers into
50, and both the estuaries were not found to be in the area which is evident due to the high nitrate and
this category in any of the seasons (Fig. 4). During BOD values during this season and the further drop
the pre-monsoon season, the WQI ranged from 100 in the WQI in the pre-monsoon season in both Man-
to 130 (Fig. 4), indicating poor water quality as the dovi and Zuari estuaries. This can be associated with
standard limit of WQI from 100 to 200 is catego- the dumping of agricultural wastes as reflected in the
rised as ‘poor’. Though most of the physicochemical elevated levels of essential nutrients like nitrate and
parameters in the pre-monsoon season were within phosphate. The abundant nutrient supply encour-
limits, a wide difference in major parameters like ages further growth of microbial flora accompanied
BOD and nitrate was observed. The values of tem- by increased oxygen demands. Another possible fac-
perature, salinity and phosphate were also very close tor contributing to this is extensive tourism activities
to the permissible limits and clearly contributed to along the coastal area, especially during November to
the reduction in WQI in the estuaries during the pre- February. WQI serves to improve the understanding
monsoon season. The WQI was greatly improved in of water quality issues by generating a score describ-
the monsoon season as all the physical parameters ing water quality status and aids in evaluating the
analysed were within the permissible range and all water quality trends (Cude, 2001; Hoorman et al.,
the values showed improved water quality. The WQI 2008).
values between 50 and 100 which are considered
to be ‘good’ waters were observed only during the Assessment of microbial pollution: seasonal
monsoon season (Fig. 4). This could be due to the variations in TVC and PIOs
increased river runoff and flushing. Earlier studies
also showed that increased freshwater runoff dur- Estuarine waters have wide variations in the physi-
ing monsoon season dilutes the concentration of the cal and chemical parameters; especially the nutrient
contaminants and removes them out of the estuarine concentration and the organic matter composition
system (VishnuRadhan et al., 2015). can influence the microbial load and its community

Fig. 4  Water quality index 350

in the Mandovi (white) and Unfit for usage
Zuari (grey) estuaries 300
Water Quality Index (WQI)

250
Very Poor
200

150
Poor
100
Good
50
Excellent
0
Pre-monsoon Monsoon Post-monsoon

Vol:. (1234567890)
13
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
Environ Monit Assess (2022) 194: 71 Page 11 of 17 71

composition. Apart from it, the inflow of waters 1.0E+08

(a)
from different terrestrial habitats brings in various
microbes of allochthonous origin. They also have an 1.0E+06

CFUs ml-1 / g-1

influence on the composition of the natural bacterial
community, and changes in the native bacterial load 1.0E+04
can be seen (Khandeparker et al., 2015). If the natural
balance is not established, there are instances when
1.0E+02
the terrestrial population can replace the estuarine
community and alter the delicate balance which can
lead to an outbreak of pathogenic bacterial communi- 1.0E+00
Pre-monsoon Monsoon Post-monsoon
ties. The occurrence of PIOs in the samples reflects a
higher incidence of sewage indicator bacteria in any 1.0E+08 (b)
environment (Fujioka & Yoneyama, 2002). Assess- Surface
ing the levels of pollution indicators are essential to Bottom
1.0E+06 Sediment
understand the microbial pollutants present in the

CFUs ml-1 / g-1

Mandovi and Zuari estuaries.
The total bacterial counts were determined based 1.0E+04
on the TVC, and the highest counts were seen dur-
ing the pre-monsoon and monsoon seasons with ~ ­104 1.0E+02
to ­106 CFU/ml in the water and sediment sam-
ples. However, the microbial load was only ~ ­102 to
­103 CFU/ml during the post-monsoon season in both
1.0E+00
Pre-monsoon Monsoon Post-monsoon
the estuaries (Fig. 5). The levels of PIOs, TC, ECLO,
SFLO, SALO, PALO, SLO, SHLO, KALO, PVLO Fig. 5  Total viable counts in (a) Mandovi and (b) Zuari estu-
and VLO, were estimated in the water and sediment aries; error bars represent standard deviation (n = 8)
samples (Table 4). TC, SFLO and SALO with maxi-
mum numbers of ~ ­102 CFUs were present only dur-
ing monsoon, and the numbers were below detectable
limits in the pre- and post-monsoon seasons. The PIO The occurrence of PIOs in the samples reflects
counts in the water and sediment samples relate to an the presence of microbial pollution in the ecosys-
increase in faecal contamination around the sampling tem, which directly reflects the presence of patho-
site due to river runoff. Maximum PALO numbers of genic bacteria that can affect the people who are
­104–106 CFU/ml were recorded in the pre-monsoon exposed to these habitats (Fujioka & Yoneyama,
period, and the numbers during the rains and in the fol- 2002). Earlier studies on the counts of pollution
lowing post-monsoon season numbers were less than indicator bacteria have also concluded that urban
100 CFU/ml. SHLO was also found to be maximum sewage outfalls impact the water quality in these
with ­102–104 CFU/ml in the pre-monsoon season, and regions due to the mixing of waters from various
the numbers were 5 to 300 CFU/ml in the monsoon point and non-point sources into the Mandovi and
and after the rains in post-monsoon. The highest VLO Zuari estuarine waters (Rodrigues et al., 2011).
counts were also recorded in the pre-monsoon period, Periodic investigations on the levels of PIOs and
with ­103–105 CFUs, and in the other two seasons, it their distribution are required for efficient manage-
was ~ 1 to 120 CFUs. This could be due to anthropo- ment of the effluent outfalls into these delicate estu-
genic contamination which is increased during the arine systems. Protecting the ecological health of
pre-monsoon season due to increased evaporation and these habitats can be achieved only based on these
reduced runoff (Table 4). PIOs such as ECLO, SLO, scientific studies for developing stringent guidelines
PVLO and KALO were below detectable levels in all and appropriate policies. These can provide appro-
three seasons. Seasonal changes in the levels of TC, priate methods for reducing the detrimental impact
SALO, PALO, SHLO and VLO were supported with of microbial pollution on human health (Nagvenkar
statistical significance (Table 3). & Ramaiah, 2009).

Vol.: (0123456789)
13
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
 71 Page 12 of 17 Environ Monit Assess (2022) 194: 71

Table 4  Average ± standard deviation (n = 8) abundance of pollution indicator organisms (CFU ­ml−1 /g−1) in the Mandovi and Zuari
estuaries
TC SFLO SALO PALO SHLO VLO
MANDOVI

Surface water ND 0±0 0±0 215 ± ­17a 344 ± 122 116 ± ­13a
Pre-monsoon season Bottom water ND 3±1 0±0 39 ± ­9a 873 ± 84 18 ± ­4a
Sediment ND 2±3 0±0 4565 ± ­18a 1179 ± 120 3044 ± ­184a
Surface water 15 ± 12 128 ± 167 1±0 1±2 316 ± 439 3±2
Monsoon season Bottom water 11 ± 2 230 ± 186 1±2 4±5 393 ± 551 4±4
Sediment 5±4 141 ± 64 367 ± 75 1±1 2±0 4±4
Surface water 2±2 0±0 1±1 2±3 23 ± 11 6±5
Post-monsoon season Bottom water 1±1 12 ± 1 1±0 84 ± 113 15 ± 20 3±2
Sediment 1±1 5±2 8±5 4±6 18 ± 17 114 ± 50
ZUARI
Surface water ND 0±0 0±0 23 ± ­4a 128 ± 42 226 ± ­12a
Pre-monsoon season Bottom water ND 7±3 0±0 45 ± ­15a 59 ± 3 303 ± ­21a
Sediment ND 446 ± 316 0±0 18,214 ± ­1092a 220 ± 311 4723 ± ­217a
Surface water 1±2 6±3 4±3 1±1 17 ± 4 5±6
Monsoon season Bottom water 1±2 7±1 4±4 24 ± 34 3±2 2±1
Sediment 177 ± 251 63 ± 44 146 ± 17 1±2 6±8 16 ± 15
Surface water 0±0 2±1 1±1 1±0 18 ± 22 2±2
Post-monsoon season Bottom water 0±0 1±1 1±0 36 ± 25 6±6 6±8
Sediment 3±1 17 ± 6 24 ± 4 3±1 17 ± 16 74 ± 98

ND not determined, TC total coliforms, SFLO Streptococcus faecalis-like organisms, SALO Staphylococcus aureus-like organisms,
PALO Pseudomonas aeruginosa-like organisms, SHLO Shigella-like organisms, VLO Vibrio-like organisms
a
Values represented are higher by order of ­102

Characterisation of antibiotic resistivity in PIOs: PIOs. Isolates with an MAR index of > 0.4 accounted
estimation of MAR index for about 5.6% which included isolate PR13 and PR27
belonging to the Vibrio and Shigella group of PIOs
Approximately 90 representative isolates of major pol- (Table 6). Enumerating the MAR index proves to be a
lution indicator bacterial groups from all three seasons useful method for health risk assessment, and isolates
were tested for their susceptibility towards 16 differ- having values > 0.2 suggest it as possible means of con-
ent antibiotics belonging to bacteriostatic and bacte- tamination which could be categorised as ‘high risk’
ricidal classes. Among the 35 isolates tested from the (Davis & Brown, 2016). Similar evidence of partial or
pre-monsoon season, 30 isolates showed resistance complete susceptibility for multiple drugs in pathogens
to Clindamycin, while all of them were susceptible to has also been reported earlier in other coastal waters,
Ofloxacin and Tetracycline (Supplementary Table S1). and in most cases, contamination was caused by urban
About 86% of the isolates tested showed an MAR index wastes (Austin et al., 2009; Dang et al., 2008; Henriques
of < 0.2 (Table 5) which can be categorised as low-risk et al., 2008).

Table 5  Percentage Category MAR index Pre-monsoon Monsoon Post-monsoon

frequency of isolates with
varying multiple antibiotic Low risk < 0.2 86.1 75.8 68
resistance (MAR) indices
High risk 0.2–0.4 8.3 15.2 28
Very high risk > 0.4 5.6 9.1 4

Vol:. (1234567890)
13
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
Environ Monit Assess (2022) 194: 71 Page 13 of 17 71

Table 6  Pollution indicator organisms from Mandovi and Zuari estuaries with very high multiple antibiotic resistance (MAR) indi-
ces
Pre-monsoon Monsoon Post-monsoon
a
Isolates PR13 PR27 M2 M7 M20 PT13
Vibrio Shigella Vibrio Coliform Coliform Vibrio
Class of antibiotic Antibiotic tested (µg) Diameter of growth inhibition zone (mm)
Bacteriostatic agents
Chloramphenicol Chloramphenicol (30) 0 0 19 0 15 14
Tetracyclines Tetracycline (30) 21 16 15 0 9 7
Macrolides Azithromycin (15) 0 29 0 0 0 0
Sulfonamides Co-trimoxazole (25) 21 28 10 0 10 0
Bactericidal agents
Ciprofloxacin (5) 0 0 14 0 16 14
Quinolones Nalidixic acid (30) 10 0 0 0 12 0
Ofloxacin (5) 14 16 11 0 11 0
Lincosamides Clindamycin (2) 0 0 0 8 7 0
Aminoglycoside Netillin (30) 17 20 10 0 8 18
Ansamycins Rifampicin (5) 0 0 0 0 0 0
Glycopeptides Vancomycin (30) 0 0 0 0 0 0
Ampicillin (10) 0 16 0 0 0 0
Ceftizoxime (30) 0 22 0 0 0 17
β Lactam Ceftriaxone (30) 0 27 0 0 0 21
Piperacillin (100) 18 0 0 0 0 0
Ertapenem (10) 15 35 19 0 12 20
% Resistance 56.3 43.8 56.25 93.75 43.75 56.25
MAR index 0.56 0.44 0.56 0.94 0.44 0.56
A complete list of pollution indicator organisms tested are provided in Supplementary Table S1
PR pre-monsoon, M monsoon, PT post-monsoon
a
Representative isolates with MAR index of > 0.4

During the monsoon sampling, a total of 33 iso- population depends on the estuaries for food and rec-
lates were screened and a coliform isolate from Man- reational activities.
dovi estuary (M7) showed resistance to all the anti- Such studies on antibiotic resistance in indige-
biotics tested except Clindamycin, which appeared nous waters are therefore important, as they indicate
with a very small inhibition zone of 8 mm only the extent of human alteration of water ecosystems
(Table 6). Approximately 75.8% of the isolates exhib- (Baquero et al., 2008). An increase in a load of path-
ited an MAR index value < 0.2, and 9.1% had MAR ogenic bacteria leads to an alteration of the natural
values of > 0.4 (Table 5). This includes isolates M2, microbiota, and the strains resistant to commonly used
M7 and M20 exhibiting resistance to 9, 15 and 7 antibiotics can be a major threat to those involved in
antibiotics and MAR index of 0.56, 0.94 and 0.44, fishing and recreational activities in these regions
respectively. The intensity of resistance, shown by (Ramaiah et al., 2004). Most of the isolates showed
these isolates during this season, is alarmingly high. resistance to the ß lactam, Lincosamide, Glycopep-
Antibiotic resistance in the post-monsoon was fairly tide and Quinolone classes of antibiotics. Therefore,
low, with only one isolate (PT13) exhibiting maxi- resistance strategies that prevent the inhibition of cell
mum resistance to 9 antibiotics with an MAR index wall biosynthesis, preventing DNA replication and
of 0.56 (Tables 5 and 6). Isolates with MAR indi- inhibition of transcription, are likely the major pro-
ces > 0.4 pose a great threat as a majority of the local cesses by which these isolates developed resistance

Vol.: (0123456789)
13
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
 71 Page 14 of 17 Environ Monit Assess (2022) 194: 71

to these antibiotics. Understanding the mechanism Mandovi and Zuari estuaries are well ventilated
by which the bacteria acquire resistance can provide by the annual monsoonal rains, and the water qual-
better treatment options. Such studies might also help ity status is maintained by the increased river runoff
detect radical changes in terms of antibiotic resistance each year. However, this study emphasises the need
of emergent pathogens of the marine system in order to study not only the physicochemical parameters and
to alert the government, public and social institutions the microbial numbers but also the pathogenicity,
of their implications. Calculation of the MAR index virulence and antibiotic sensitivity of the PIOs pre-
provides a better understanding of the virulence of sent in the habitat. Any reduction in the monsoonal
PIOs and serves as an excellent tool for health risk rains and an increase in the number of virulent PIOs
assessment (Chitanand et al., 2010). Though in this entering the estuarine habitat can alter the microbial
study, the presence of antibiotic residues was not car- community structure. Hence, adequate treatment of
ried out, its influence is clearly evident based on the urban runoff and terrestrial inputs into the estuarine
natural resistance observed in bacterial communities regions has to be undertaken. Studies on the expres-
from these regions. Studies have also shown that the sion of virulence genes by pathogens in the estuary
increased density of resistant bacteria and the pres- need to be taken up to ensure the long-term safety
ence of favourable growth conditions for their prolif- of those dependent on the estuarine habitat for their
eration can lead to higher health risks (Diwan et al., livelihood. Such studies can play an important role
2010). to frame policies for sustainable development and
conservation of these ecologically important coastal
zones.
Conclusion

In this study, the seasonal changes of the physico-

chemical parameters and microbial load were deter- Supplementary information
mined in the Mandovi and Zuari estuaries. The water
quality index was computed to understand the current Supplementary Table S1 Multiple antibiotic resist-
environmental status of the estuaries. Results show ance (MAR) index of pollution indicator organisms
that the overall water quality conditions remain con- isolated from pre-monsoon, monsoon and post-mon-
siderably good due to the annual southwest monsoon. soon seasons, associated with this article is provided
It has a positive impact on the major physicochemical in the file ‘Supplementary Material.xlsx’.
parameters such as temperature, salinity and dissolved
Acknowledgements The authors are thankful to the direc-
oxygen levels. Studies on the microbial load and the
tor of CSIR-National Institute of Oceanography, Goa, for
levels of PIOs especially based on CFUs also sug- extending all the support and required facilities to accomplish
gest that a load of bacteria is increased in the pre- and this work. Research funding to study the seawater quality in
post-monsoon periods and the numbers are reduced the Mandovi and Zuari estuarine system from the Ministry of
Earth Sciences, Govt. of India, is gratefully acknowledged.
during the monsoon period. However, the MAR indi-
Thanks to all other project assistants who participated in sea-
ces determined for the different PIOs suggest that the sonal fieldwork and helped in sample collection. This is CSIR-
water quality is not completely safe during the mon- NIO contribution No. 6845.
soon season due to increased terrestrial load, which
could bring in PIOs with a very high MAR index. Funding Ministry of Earth Sciences, Government of India,
under the research programme, MoES/ICM AM-PD/Suppl.
Bacteria with > 0.2 MAR index are considered to
Order/81/2017.
be highly detrimental to the health of the individu-
als exposed to these microbes. In this study, we have Availability of data and material Not applicable.
observed that during the monsoon season, an isolate
with the highest MAR index of 0.94 was obtained. Code availability Not applicable.
This shows that the seasonal changes along with the
Declarations
increased terrestrial load from different sources can
alter the water quality in the estuarine region and the Conflict of interest The authors declare no competing inter-
virulence of the microbes in these habitats. ests.

Vol:. (1234567890)
13
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
Environ Monit Assess (2022) 194: 71 Page 15 of 17 71

References coastal waters of Jiaozhou Bay. Microbial Ecology., 55(2),

237–246. https://​doi.​org/​10.​1007/​s00248-​007-​9271-9
Adamou, H., Ibrahim, B., Salack, S., Adamou, R., Sanfo, S., Davis, R., & Brown, P. D. (2016). Multiple antibiotic resistance
Liersch, S. (2020). Physico-chemical and bacteriological index, fitness and virulence potential in respiratory Pseu-
quality of groundwater in a rural area of Western Niger: domonas aeruginosa from Jamaica. Journal of Medical
A case study of Bonkoukou. Journal of Water and Health, Microbiology, 65(4), 261–271. https://​doi.​org/​10.​1099/​
18(1), 77–90. jmm.0.​000229
Alagarsamy, R. (2006). Distribution and seasonal variation of De Souza, M. J., Nair, S., Bharathi, P. L., & Chandramohan,
trace metals in surface sediments of the Mandovi estuary, D. (2006). Metal and antibiotic-resistance in psychro-
west coast of India. Estuarine, Coastal and Shelf Science, trophic bacteria from Antarctic Marine waters. Eco-
67(1–2), 333–339. https://​doi.​org/​10.​1016/j.​ecss.​2005.​11.​ toxicology, 15(4), 379–384. https://​doi.​org/​10.​1007/​
023 s10646-​006-​0068-2
Alam, M. J., Islam, M. R., Muyen, Z., Mamun, M., & Islam, Dheenan, P. S., Jha, D. K., Vinithkumar, N. V., Ponmalar, A.
S. (2007). Water quality parameters along rivers. Interna- A., Venkateshwaran, P., & Kirubagaran, R. (2014). Spa-
tional Journal of Environmental Science and Technology, tial variation of physicochemical and bacteriological
4(1), 159–167. parameters elucidation with GIS in Rangat Bay, Middle
Austin, C. B., McArthur, J. V., Lindell, A. H., Wright, M. Andaman. India. Journal of Sea Research., 85, 534–541.
S., Tuckfield, R. C., Gooch, J., Warner, L., Oliver, J., & https://​doi.​org/​10.​1016/j.​seares.​2013.​09.​001
Stepanauskas, R. (2009). Multi-site analysis reveals wide- Diwan, V., Tamhankar, A. J., Khandal, R. K., Sen, S., Aggarwal,
spread antibiotic resistance in the marine pathogen Vibrio M., Marothi, Y., Iyer, R. V., Sundblad-Tonderski, K., &
vulnificus. Microbial Ecology, 57, 151–159. https://​doi.​ Stålsby-Lundborg, C. (2010). Antibiotics and antibiotic-
org/​10.​1007/​s00248-​008-​9413-8 resistant bacteria in waters associated with a hospital in
Avvannavar, S. M., & Shrihari, S. (2008). Evaluation of water Ujjain. India. BMC Public Health, 10(1), 414. https://​doi.​
quality index for drinking purposes for river Netravathi, org/​10.​1186/​1471-​2458-​10-​414
Mangalore. South India. Environmental Monitoring and Fondriest Environmental (2013, November 19). Dissolved
Assessment, 143(1–3), 279–290. https://​doi.​org/​10.​1007/​ oxygen: Fundamentals of environmental measurements.
s10661-​007-​9977-7 Retrieved December 10, 2020, from https://​www.​fondr​iest.​
Baquero, F., Martínez, J. L., Cantón, R. (2008). Antibiotics and com/​envir​onmen​tal-​measu​remen​ts/​param​eters/​water-​quali​ty/​
antibiotic resistance in water environments. Current Opin- disso​lved-​oxygen/
ion in Biotechnology, 19(3), 260–265. Fujioka, R. S., Yoneyama, B. S. (2002). Sunlight inactivation
Bauer, A. W. (1966). Antibiotic susceptibility testing by a of human enteric viruses and fecal bacteria. Water Science
standardized single disc method. American Journal of and Technology, 46(11–12), 291–295. https://​doi.​org/​10.​
Clinical Pathology, 45, 149–158. NII Article ID (NAID) 2166/​wst.​2002.​0752
10024190092 Grasshoff, K. (1983). Determination of nutrients. Methods of
Bhavya, P. S., Kumar, S., Gupta, G. V. M., Sudheesh, V., seawater analysis, 125–187.
Sudharma, K. V., Varrier, D. S., Dhanya, K. R., & Gupta, A. K., Gupta, S. K., & Patil, R. S. (2003). A compari-
Saravanane, N. (2016). Nitrogen uptake dynamics in a son of water quality indices for coastal water. Journal of
tropical eutrophic estuary (Chochin, India) and adjacent Environmental Science and Health. Part A, 38(11), 2711–
coastal waters. Estuaries and Coasts., 39, 54–67. https://​ 2725. https://​doi.​org/​10.​1081/​ESE-​12002​4458
doi.​org/​10.​1007/​s12237-​015-​9982-y Henriques, I. S., Fonseca, F., Alves, A., Saavedra, M. J., &
Carey, J. C., & Fulweiler, R. W. (2014). Salt marsh tidal Correia, A. (2008). Tetracycline-resistance genes in
exchange increases residence time of silica in estuaries. Gram-negative isolates from estuarine waters. Letters in
Limnology and Oceanography., 59(4), 1203–1212. https://​ Applied Microbiology, 47(6), 526–533. https://​doi.​org/​10.​
doi.​org/​10.​4319/​lo.​2014.​59.4.​1203 1111/j.​1472-​765x.​2008.​02452.x
Chakraborty, P. (2017). Oyster reef restoration in controlling Hocking, M. B. (2005). Water quality measurement. In M. B.
coastal pollution around India: A viewpoint. Marine Pol- Hocking (Ed.), Handbook of Chemical Technology and
lution Bulletin. 115(1–2), 190–193. Pollution Control (3rd ed., pp. 105–135). Elsevier Inc.
Chitanand, M. P., Kadam, T. A., Gyananath, G., Totewad, N. Hoorman, J., Hone, T., Sudman, T., Dirksen, T., Iles, J.,
D., Balhal, D. K. (2010). Multiple antibiotic resistance Islam, K. R. (2008). Agricultural impacts on lake and
indexing of coliforms to identify high risk contamina- stream water quality in Grand Lake St. Marys, Western
tion sites in aquatic environment. Indian Journal of Ohio. Water, Air, and Soil Pollution, 193(1–4), 309–322.
Microbiology, 50(2), 216–220. https://​doi.​org/​10.​1007/​ https://​doi.​org/​10.​1007/​s11270-​008-​9692-1
s12088-​010-​0042-9 Jayakumar, D. A., Francis, C. A., Naqvi, S. W. A., Ward, B.
Cude, C. G. (2001). Oregon water quality index a tool for eval- B. (2004). Diversity of nitrite reductase genes (nirS) in
uating water quality management effectiveness. JAWRA the denitrifying water column of the coastal Arabian
Journal of the American Water Resources Association, Sea. Aquatic Microbial Ecology, 34(1), 69–78. https://​doi.​
37(1), 125–137. https://​doi.​org/​10.​1111/j.​1752-​1688.​ org/​10.​3354/​ame03​4069
2001.​tb054​80.x Jha, D. K., Devi, M. P., Vidyalakshmi, R., Brindha, B., Vinithkumar,
Dang, H., Ren, J., Song, L., Sun, S., & An, L. (2008). Diverse N. V., & Kirubagaran, R. (2015). Water quality assessment
tetracycline resistant bacteria and resistance genes from using water quality index and geographical information

Vol.: (0123456789)
13
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
 71 Page 16 of 17 Environ Monit Assess (2022) 194: 71

system methods in the coastal waters of Andaman Sea. India. Monitoring and Assessment, 155(1–4), 245. https://​doi.​
Marine Pollution Bulletin, 100(1), 555–561. https://​doi.​org/​ org/​10.​1007/​s10661-​008-​0432-1
10.​1016/j.​marpo​lbul.​2015.​08.​032 Qasim, S. Z., & Gupta, R. S. (1981). Environmental charac-
Kannel, P. R., Lee, S., Lee, Y. S., Kanel, S. R., Khan, S. P. teristics of the Mandovi-Zuari estuarine system in Goa.
(2007). Application of water quality indices and dis- Estuarine, Coastal and Shelf Science, 13(5), 557–578.
solved oxygen as indicators for river water classification https://​doi.​org/​10.​1016/​S0302-​3524(81)​80058-8
and urban impact assessment. Environmental Monitor- Ram, A. P., Nair, S., Chandramohan, D. (2003). Seasonal
ing and Assessment. 132(1–3), 93–110. https://​doi.​org/​ shift in net ecosystem production in a tropical estu-
10.​1007/​s10661-​006-​9505-1 ary. Limnology and Oceanography, 48(4), 1601–1607.
Kessarkar, P. M., Rao, V. P., Shynu, R., Mehra, P., & Viegas, https://​doi.​org/​10.​4319/​lo.​2003.​48.4.​1601
B. E. (2010). The nature and distribution of particulate Ramaiah, N., Kolhe, V., Sadhasivan, A. (2004). Abundance
matter in the Mandovi estuary, central west coast of of pollution indicator and pathogenic bacteria in Mum-
India. Estuaries and Coasts., 33(1), 30–44. https://​doi.​ bai waters. Current Science, 87(4), 435–439.
org/​10.​1007/​s12237-​009-​9226-0 Rodrigues, V., Ramaiah, N., Kakti, S., & Samant, D. (2011).
Khandeparker, L., Anil, A. C., Naik, S. D., & Gaonkar, C. C. Long-term variations in abundance and distribution of
(2015). Daily variations in pathogenic bacterial popu- sewage pollution indicator and human pathogenic bac-
lations in a monsoon influenced tropical environment. teria along the central west coast of India. Ecological
Marine Pollution Bulletin., 96(1–2), 337–343. https://​ Indicators, 11(2), 318–327. https://​doi.​org/​10.​1016/j.​
doi.​org/​10.​1016/j.​marpo​lbul.​2015.​04.​051 ecoli​nd.​2010.​05.​010
Kumar, B. S. K., & Sarma, V. V. S. S. (2018). Variations in Şener, Ş, Şener, E., & Davraz, A. (2017). Evaluation of water
concentrations and sources of bioavailable organic com- quality using water quality index (WQI) method and
pounds in the Indian estuaries and their fluxes to coastal GIS in Aksu River (SW-Turkey). Science of the Total
waters. Continental Shelf Research, 166, 22–33. https://​ Environment, 584, 131–144. https://​doi.​org/​10.​1016/j.​
doi.​org/​10.​1016/j.​csr.​2018.​07.​001 scito​tenv.​2017.​01.​102
Lin, J., Xie, L., Pietrafesa, L. J., Shen, J., Mallin, M. A., & Sharpley, A. N., McDowell, R. W., & Kleinman, P. J. (2001).
Durako, M. J. (2006). Dissolved oxygen stratification Phosphorus loss from land to water: Integrating agricultural
in two micro-tidal partially-mixed estuaries. Estuarine, and environmental management. Plant and Soil, 237(2),
Coastal and Shelf Science, 70(3), 423–437. https://​doi.​ 287–307. https://​doi.​org/​10.​1023/A:​10133​35814​593
org/​10.​1016/j.​ecss.​2006.​06.​032 Shetye, S. R., Shankar, D., Neetu, S., Suprit, K., Michael, G.
Liu, B., Wang, W. L., Han, R. M., Sheng, M., Ye, L. L., Du, S., & Chandramohan, P. (2007). The environment that
X., Wu, X. T., & Wang, G. X. (2016). Dynamics of dis- conditions the Mandovi and Zuari estuaries. In S. R.
solved oxygen and the affecting factors in sediment of Shetye, D. Kumar, & D. Shankar (Eds.), The Mandovi
polluted urban rivers under aeration treatment. Water, and Zuari Estuaries (pp. 3–26). National Institute of
Air and Soil Pollution, 227(6), 172. https://​doi.​org/​10.​ Oceanography.
1007/​s11270-​016-​2869-0 Shynu, R., Rao, V. P., Sarma, V. V. S. S., Kessarkar, P. M.,
Maloo, A., Fulke, A. B., Mulani, N., Sukumaran, S., & Ram, Mani Murali, R. (2015). Sources and fate of organic
A. (2017). Pathogenic multiple antimicrobial resist- matter in suspended and bottom sediments of the Man-
ant Escherichia coli serotypes in recreational waters of dovi and Zuari estuaries, western India. Current Sci-
Mumbai, India: A potential public health risk. Environ- ence, 108(2); 226–238. https://​www.​jstor.​org/​stable
mental Science and Pollution Research, 24(12), 11504– /24218162
11517. https://​doi.​org/​10.​1007/​s11356-​017-​8760-8 Sundar, D., Shetye, S. R. (2005). Tides in the Mandovi and
Manoj, N. T., Unnikrishnan, A.S., Sundar, D. (2009) Tidal Zuari estuaries, Goa, west coast of India. Journal of
asymmetry in the Mandovi and Zuari estuaries, the west Earth System Science, 114(5), 493–503.
coast of India Journal of Coastal Research, 1187–1197. USEPA (1986). Ambient water quality criteria for bacteria
https://​doi.​org/​10.​2112/​08-​1056.1 (Washington: U.S. Environmental Protection Agency,
Maya, M. V., Soares, M. A., Agnihotri, R., Pratihary, A. K., EPA440/5–84–002).
Karapurkar, S., Naik, H., & Naqvi, S. W. A. (2011). Vignesh, S., Dahms, H. U., Emmanuel, K. V., Gokul, M. S.,
Variations in some environmental characteristics includ- Muthukumar, K., Kim, B. R., & James, R. A. (2014).
ing C and N stable isotopic composition of suspended Physicochemical parameters aid microbial commu-
organic matter in the Mandovi estuary. Environmental nity? A case study from marine recreational beaches.
Monitoring and Assessment, 175, 501–517. https://​doi.​ Southern India. Environmental Monitoring and Assess-
org/​10.​1007/​s10661-​010-​1547-8 ment, 186(3), 1875–1887. https://​doi.​org/​10.​1007/​
Moss, J. A., Nocker, A., Lepo, J. E., & Snyder, R. A. (2006). s10661-​013-​3501-z
Stability and change in estuarine biofilm bacterial com- Vijith, V., Sundar, D., Shetye, S. R. (2009). Time-dependence
munity diversity. Applied and Environmental Microbiol- of salinity in monsoonal estuaries. Estuarine, Coastal
ogy, 72(9), 5679–5688. https://​doi.​org/​10.​1128/​AEM.​ and Shelf Science, 85(4), 601–608.
02773-​05 VishnuRadhan, R., Sagayadoss, J., Seelan, E., Vethamony,
Nagvenkar, G. S., & Ramaiah, N. (2009). Abundance of P., Shirodkar, P., Zainudin, Z., & Shirodkar, S. (2015).
sewage-pollution indicator and human pathogenic bac- Southwest monsoon influences the water quality and
teria in a tropical estuarine complex. Environmental waste assimilative capacity in the Mandovi estuary (Goa

Vol:. (1234567890)
13
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
Environ Monit Assess (2022) 194: 71 Page 17 of 17 71

state, India). Chemistry and Ecology, 31(3), 217–234. Publisher’s Note Springer Nature remains neutral with regard
https://​doi.​org/​10.​1080/​02757​540.​2014.​961435 to jurisdictional claims in published maps and institutional
Yang, X., Wu, X., Hao, H., & He, Z. (2008). Mechanisms affiliations.
and assessment of water eutrophication. Journal of Zhe-
jiang University Science B, 9(3), 197–209. https://​doi.​
org/​10.​1631/​jzus.​B0710​626

Vol.: (0123456789)
13
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
Terms and Conditions
Springer Nature journal content, brought to you courtesy of Springer Nature Customer Service Center GmbH (“Springer Nature”).
Springer Nature supports a reasonable amount of sharing of research papers by authors, subscribers and authorised users (“Users”),
for small-scale personal, non-commercial use provided that all copyright, trade and service marks and other proprietary notices are
maintained. By accessing, sharing, receiving or otherwise using the Springer Nature journal content you agree to these terms of use
(“Terms”). For these purposes, Springer Nature considers academic use (by researchers and students) to be non-commercial.
These Terms are supplementary and will apply in addition to any applicable website terms and conditions, a relevant site licence or
a personal subscription. These Terms will prevail over any conflict or ambiguity with regards to the relevant terms, a site licence or
a personal subscription (to the extent of the conflict or ambiguity only). For Creative Commons-licensed articles, the terms of the
Creative Commons license used will apply.
We collect and use personal data to provide access to the Springer Nature journal content. We may also use these personal data
internally within ResearchGate and Springer Nature and as agreed share it, in an anonymised way, for purposes of tracking,
analysis and reporting. We will not otherwise disclose your personal data outside the ResearchGate or the Springer Nature group of
companies unless we have your permission as detailed in the Privacy Policy.
While Users may use the Springer Nature journal content for small scale, personal non-commercial use, it is important to note that
Users may not:

1. use such content for the purpose of providing other users with access on a regular or large scale basis or as a means to
circumvent access control;
2. use such content where to do so would be considered a criminal or statutory offence in any jurisdiction, or gives rise to civil
liability, or is otherwise unlawful;
3. falsely or misleadingly imply or suggest endorsement, approval , sponsorship, or association unless explicitly agreed to by
Springer Nature in writing;
4. use bots or other automated methods to access the content or redirect messages
5. override any security feature or exclusionary protocol; or
6. share the content in order to create substitute for Springer Nature products or services or a systematic database of Springer
Nature journal content.
In line with the restriction against commercial use, Springer Nature does not permit the creation of a product or service that creates
revenue, royalties, rent or income from our content or its inclusion as part of a paid for service or for other commercial gain.
Springer Nature journal content cannot be used for inter-library loans and librarians may not upload Springer Nature journal
content on a large scale into their, or any other, institutional repository.
These terms of use are reviewed regularly and may be amended at any time. Springer Nature is not obligated to publish any
information or content on this website and may remove it or features or functionality at our sole discretion, at any time with or
without notice. Springer Nature may revoke this licence to you at any time and remove access to any copies of the Springer Nature
journal content which have been saved.
To the fullest extent permitted by law, Springer Nature makes no warranties, representations or guarantees to Users, either express
or implied with respect to the Springer nature journal content and all parties disclaim and waive any implied warranties or
warranties imposed by law, including merchantability or fitness for any particular purpose.
Please note that these rights do not automatically extend to content, data or other material published by Springer Nature that may be
licensed from third parties.
If you would like to use or distribute our Springer Nature journal content to a wider audience or on a regular basis or in any other
manner not expressly permitted by these Terms, please contact Springer Nature at

onlineservice@springernature.com