Bull Environ Contam Toxicol (2017) 99:23–26
DOI 10.1007/s00128-017-2110-x
Effects of Water pH on Life History Parameters of a New
Bosminid Cladocera: Bosmina (Bosmina) Tripurae (Korinek, Saha
and Bhattachaya, 1999) in Laboratory Condition
Himadri Saha1   · K. S. Wisdom1 · Asem Lembika Devi1 · Da‑u‑ruhi Pde1 ·
Sukham Tuni Devi1 · Makamguang Kamei1 · Anamika Biswas1 · Ratan K. Saha1 ·
Prasenjit Pal1 
Received: 29 March 2016 / Accepted: 9 May 2017 / Published online: 19 May 2017
© Springer Science+Business Media New York 2017
Abstract  The effects of water pH on life history param-
eters of Bosmina tripurae have been studied to determine
the most suitable water pH desired for the maximum
growth and development of this newly discovered clad-
oceran species. The study was carried out under the labo-
ratory condition at 20 ± 2°C. Five pH ranges 5.0 ± 0.2,
6.0 ± 0.2, 7.0 ± 0.2, 8.0 ± 0.2 and 9.0 ± 0.2 with six rep-
licates for each pH consisting of one animal in each Petri
dish (80 × 15  mm) were used for the study. 20  mL of
respective test medium was maintained with Chlorella sp.
(2 × 104 ± 0.03  cells  mL−1) in each Petri dish throughout
the experiment. Thirty (30) animals were observed daily
to investigate different life history parameters like total life
span, age at maturity, number of eggs, neonates and egg
batches etc. at different condition. From the study it was
found that acidic water (pH 5 ± 0.2) is more suitable for the
culture of Bosmina tripurae in laboratory condition.
Keywords Cladocera · Bosmina tripurae · Water pH ·
Eggs · Neonates · Hatching · Embryonic development
Cladocerans are most abundant and major components of
zooplankton in many aquatic ecosystems. They have spe-
cial role in transfer of energy to the higher trophic levels
such as juvenile fishes and aquatic invertebrates (Santos
et al. 2010) and thus they act as a bridge between primary
producer and other consumers in the natural food-web.
* Himadri Saha
sahacofcau@gmail.com
1 ism in ecotoxicological bioassay, as it is abundant in fresh-
Department of Aquatic Health and Environment, College
of Fisheries, Central Agricultural University, Lembucherra,
West Tripura, Tripura 799 210, India
Genus Bosmina is one of the most important cladocera
that is widely distributed in freshwater bodies of tropical
and temperate regions (Threlkeld 1981; Siraj et  al. 2006;
Genung 2012). In ponds and lakes of Tripura, India Bos-
mina tripurae is commonly present-in zooplankton and is
one of the most dominant cladocerans (Biswas 2014).
Water pH is an important variable that influenced cla-
doceran communities in water bodies (Yan et  al. 1996;
Arnott et  al. 2001) and pH of 6.5–8.0 is optimal for most
of the aquatic organisms (Lawson 1997). Generally, most
of the species favour alkaline water (Duigan and Kovach
1991). However, some cladocerans, including Alona inter-
media, Chydorus ovalis, Bosmina longispina, Bosmina spp.
and Rhynchotalona falcate (Fryer 1993) are able to toler-
ate acidic water, and even demonstrated a preference for
this. The abundance of B. longirortris increased with acid
treatment in comparison to control treatment observed
by Havens and DeCosta (1985). However, little is known
about the effects of environmental parameters on life his-
tory parameters of Bosminids. Many experimental studies
have shown that water temperature and food conditions are
the most important factors affecting the life history param-
eters of Bosmina sp. (Urabe 2006; Begum et al. 2015). Bis-
was et al. (2014) studied the life history traits of B. tripurae
at laboratory culture, regarding growth and reproductive
parameters. However, effect of pH on life cycle parameters
of Bosminids particularly B. tripurae on labobaratory con-
dition have not been studied so far. The related studies have
mainly focused on the effect of acid stress on growth and
development, reproduction and survival of Daphnia sp.
(Walton et  al. 1982; Moustafa 2007; Ghazy et  al. 2011).
This is the first attempt to study B. tripurae as model organ-
water bodies of NE region of India (Korinek et  al. 1999;
Sinha 2002) and easy to culture under laboratory conditions
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24
(Biswas et  al. 2014). Therefore, the main purpose of this
study was to investigate the effects of pH on the reproduc-
tive parameters and life span of B. tripurae.
Materials and Methods
Stock culture of B. tripurae was initiated by collecting
mixed zooplankton and water sample from a pond named
S-4 (latitude 23°54.327′N and longitude 91°18.493′E),
near the College of Fisheries, Lembucherra, West Tripura
India. B. tripurae was isolated from the collected sam-
ple and stock culture was maintained in filtered water
obtained from S-4 pond in the laboratory of Department
of Aquatic Health and Environment, College of Fisheries
following Biswas et  al. (2014). Briefly, the species was
acclimatized for ten generation in laboratory conditions,
under a 12:12  h light/dark cycle, 600  lux light intensity
and at 20 ± 2°C. The pH of the pond water and the stock
culture were same i.e 7.0 ± 0.2. Algal food (Chlorella sp.
at 2 ± 0.03 × 104 cells mL−1) was provided to the stock cul-
tures as food source of B. tripurae.
Based on the pilot acute test (pH 4.0 has lethal effect),
B. tripurae was exposed to five different pH treatments
5.0 ± 0.2; 6.0 ± 0.2; 7.0 ± 0.2 (control); 8.0 ± 0.2 and
9.0 ± 0.2 with ten replicates for each treatment. Prior to
the experiment, 50 females with one egg (primiparous
female) were isolated from the stock culture and kept indi-
vidually in each Petri dish (80 × 15 mm). Neonates hatched
out from these primiparous females were used for experi-
mental study. Neonates were fed with Chlorella sp. at a
concentration of 2 ± 0.03 × 104  cells  mL−1. Chlorella sp
culture was done with Chu-10 medium (Chu 1942; FAO
1996). Observations were recorded daily to compare the
life table such as age of maturity, clutch size, number of
clutches, embryonic duration, number of neonates, and
hatching rate (Production of neonates × 100/Production of
eggs per female), life span of B. tripurae in different treat-
ments following Biswas et al. (2014). Animals which died
from each treatment within a week due to handling stress
were not included for the study. From the surviving animal,
Table 1  Comparative water quality (Mean ± SE) at different pH treatments
Treatments Water pH Temperature (°C) Dissolved oxy-
­ −1)
gen (mg L
T1 5.0 ± 0.2 20 ± 2 4.8–5.5
T2 6.0 ± 0.2 20 ± 2 5.2–5.6
T3 (Control) 7.0 ± 0.2 20 ± 2 4.3–5.6
T4 8.0 ± 0.2 20 ± 2 5.2–5.5
T5 9.0 ± 0.2 20 ± 2 5.2–5.5
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Bull Environ Contam Toxicol (2017) 99:23–26
randomly six animals were selected from each treatment
for the life history study. Experimental medium was pre-
pared as stock culture and water pH of the culture medium
for different treatments were adjusted with 1 N HCl (Hime-
dia) or 1 N NaOH (Himedia). Daily culture mediums were
renewed up to 50%. Water qualities were maintained in the
test medium for different treatments are given in Table  1.
Data were examined with one-way ANOVA and significant
difference (p < 0.05) in life table parameters in different
treatments were analyzed by Duncan’s multiple range test
using the statistical software SPSS 16.0 version.
Results and Discussion
Several studies have demonstrated that food concentration
and abiotic factors such as water temperature significantly
influenced the life histories of zooplankton (Gillooly 2000).
Hydrogen ion concentrations have a great impact on the
survival and reproduction rates of cladocera (Walton et al.
1982; Moustafa 2007; Ghazy et  al. 2011). In freshwater,
Bosmina often increases in abundance in acidified lakes,
showing the positive response to low pH. Laboratory stud-
ies about the effect of pH on survival, growth and reproduc-
tion rates of Bosmina are scanty. In the present investiga-
tion, the days of maturity was found significantly (p < 0.05)
earlier at pH 8 ± 0.2 (3.17 ± 0.60 days) as compared to pH
6 ± 0.2 and 7 ± 0.02. However, the days of maturity at pH
5 ± 0.2 and pH 9 ± 0.2 were similar to all other experimen-
tal groups. Thus in B. tripurae, days of maturity increases
with the decrease in pH which is line with Daphnia
magna (Walton et  al. 1982; Ghazy et  al. 2011). Develop-
ment time to maturity was almost same in both pH 5 ± 0.2
(4.66 ± 0.40) and at pH 9 ± 0.2 (4.25 ± 0.59) (Table 2).
Average fecundity of B. tripurae at pH 5 ± 0.2 was com-
paratively (p < 0.05) higher than the other experimental
groups. In contrary to B. tripurae, Daphnia sp. showed
decreased fecundity in lower pH (Walton et  al. 1982;
Andrea and Sprules 2000). However, many other stud-
ies have shown the significant correlation between the
mean brood size of Cladocera with food concentration and
Total alkalinity Ammonia (mg ­L−1) Light inten- Photoperiod
(mg ­L−1) sity (lux) (light and dark
hours)
103–110 0.01–0.03 600 12:12
97–103 0.01–0.02 600 12:12
100–108 0.01–0.03 600 12:12
99–108 0.01–0.03 600 12:12
92–105 0.01–0.03 600 12:12
Bull Environ Contam Toxicol (2017) 99:23–26 25

Table 2  Life cycle parameters Parameters Treatments (pH)

of B. tripurae at five different
treatments (n=6) T1 T2 T3 T4 T5
(5.0 ± 0.2) (6.0 ± 0.2) (7.0 ± 0.2) (8.0 ± 0.2) (9.0 ± 0.2)

Age at maturity (days) 5.00 ± 0.52ab 5.33 ± 0.61a 5.17 ± 0.79a 3.17 ± 0.60b 4.00 ± 0.58ab

a b b b
Mean fecundity/female 10 ± 1.18 5.33 ± 0.88 6.17 ± 1.78 3.00 ± 0.63 5.33 ± 1.20b
a b b b
No of neonates/female 9.17 ± 1.17 4.83 ± 0.70 5.17 ± 1.78 2.50 ± 0.56 3.00 ± 0.93b
a b b b
No of clutches 4.33 ± 0.33 2.33 ± 0.33 3.00 ± 0.58 2.00 ± 0.37 2.33 ± 0.21b
a a a a
Overall hatching % 92.21 ± 4.37 93.15 ± 4.80 77.99 ± 11.94 87.50 ± 8.54 51.75 ± 4.87b
a a a ab
Embryonic duration (days) 2.00 ± 0.26 1.83 ± 0.31 2.00 ± 0.68 3.17 ± 0.60 4.00 ± 0.58b
Average life span (days) 19.00 ± 0.73a 17.67 ± 0.71ab 14.17 ± 2.79bc 14.50 ± 1.36abc 12.33 ± 1.02c

Data are presented as mean  ± SE. Different superscript indicate statistically significant differences
(p < 0.05) among the experimental groups

temperature (Rose et  al. 2000). They also revealed high
pH significantly reduces the fecundity in Ceriodaphnia cf.
dubia. The present study showed that fecundity of female
decreased significantly with increase in pH from 5 ± 0.2
(Table  2). Similar observation was demonstrated in C. cf.
dubia by Belanger and Cherry (1990) that pH values above
8.99 resulted in significantly impairs reproduction. Simi-
larly, neonates production per female at pH 5 ± 0.2 were
significantly (p < 0.05) higher than the other experimental
groups and there were no significant differences (p > 0.05)
among T2, T3, T4, and T5. Thus, the neonate production
increases with the decrease in water pH.
In the present study, overall average hatching percent-
age of B. tripurae was the lowest in T5 as compared to
the other experimental groups. There was no significant
(p > 0.05) differences in the hatching rate among T1, T2,
T3 and T4 groups (Table  2). In the case of D. longspina,
high offspring number was produced at pH 8 than at pH 7
and 9 (Moustafa 2007). Accumulation of neonates depends
on hatching rate and lower production (p > 0.05) of neonate
was found at pH 8 in B. tripurae (Table 2) as compared to
other pH groups.
Average embryonic duration at pH 9 ± 0.2 (T5) was sig-
nificantly (p < 0.05) higher than the other experimental pH
groups. However, there was no significant (p > 0.05) dif-
ferences in the embryonic duration among the T1, T2, T3
and T4 groups. Thus it can be concluded that at high alka-
line pH the B. tripurae requires more time to develop. The
lifespan of T5 was significantly (p < 0.05) lower than the
T1 and T2. Thus, the lifespan of B. tripurae increases with
lowering of pH and decreases with increase in pH.
Survivability percentage of B. tripurae in different pH
treatments is shown in Fig. 1. Maximum survivability of B.
tripurae was found to be 24 days in the present experimen-
tal conditions. Biswas et al. (2014) have also reported the
total life span of B. tripurae as 24.60 ± 0.23 days. Similarly,
other species like Daphnia carinata and D. barbata has
a life span of 24 days (Navanneethakrishnan and Michael
1971) and 27.22 days (Scholtz et  al. 1988), respectively.
However, species like D. pulex had a much longer survival
time of 66.56 days (Scholtz et al. 1988). Early mortalities
were observed at pH 9 where all the animals died within
16 days of exposure. At pH 8 and 6 all the animals were
died by 20 days of exposure. However, first mortality was
observed on 15 days of exposure to pH 6 whereas at pH
8 it was on day 9. At control condition, the first mortality
was observed on the 8th day of the experiment and all the
animals died within 24 days of the experiment. However, at
pH 5, first mortality was observed on 17th day of the expo-
sure and the entire animal died by 23rd day. Similar kind of
study was performed by Ghazy et al. (2011) on D. magna
T1 T2 T3 (control) T4 T5
120
100

delay in the embryonic duration may be due to the stress 80

Survival (%)

effect imparted on the animal at high pH. 60

A number of clutches were significantly (p < 0.05) 40
higher at T1 as compared to the other experimental groups. 20
However, there was no significant (p > 0.05) differences
0
among the T2, T3, T4, and T5. Thus, it can be concluded 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
that at pH 5 ± 0.2, B. tripurae can grow very well with Days
maximum productivity.
Average life span of B. tripurae was significantly Fig. 1  Survival (%) of B. tripurae per female in five different pH
(p < 0.05) higher at T1 than the control group. However, the treatments

13

26
where they have found 100% survival rate at pH 8.33 up to
21 days of exposure. Thus, from the present study, it can
be concluded that the B. tripurae perform better at pH 5
and their survivability data is also in compliance with the
lifespan data.
It was observed that in all the cases the variable pH
have the significant effect on life history parameters of B.
tripurae. It was noted that the highest number of fecundity,
number of neonates, clutch no, hatching rate and total life
span were recorded at acidic pH. Thus, it can be concluded
that the pH of 5 ± 0.2 is most suitable for the culture of B.
tripurae.
Acknowledgements  We are grateful to Prof. M. Premjit Singh,
Hon’ble Vice-Chancellor, CAU; Dr. J. R. Dhanze, Former Dean, Col-
lege of Fisheries, CAU, Lembucherra, Tripura and Dr. P.K. Pandey,
Dean, College of Fisheries, CAU, Lembucherra, Tripura for their sup-
port and guidance to conduct the work and writing the paper.
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