Roger Williams University

DOCS@RWU

Arts & Sciences Faculty Publications Arts and Sciences

2009

Effects of temperature on reproduction and survival of the

calanoid copepod Pseudodiaptomus pelagicus
Andrew L. Rhyne
Roger Williams University, arhyne@[Link]

Cortney L. Ohs
University of Florida

Erik Stenn
AlgaGen LLC

Follow this and additional works at: [Link]

Part of the Biology Commons

Recommended Citation
Rhyne, A.L., C. Ohs, and E. Stenn. 2009. "The Effects of Temperature on Growth, Reproduction, and
Production of the Calanoid Copepod, Pseudodiaptomus Pelagicus, a Species with Potential of Mass
Production for use in Aquaculture." Aquaculture 292: 53-59.

This Article is brought to you for free and open access by the Arts and Sciences at DOCS@RWU. It has been
accepted for inclusion in Arts & Sciences Faculty Publications by an authorized administrator of DOCS@RWU. For
more information, please contact mwu@[Link].
 Aquaculture 292 (2009) 53–59

Contents lists available at ScienceDirect

Aquaculture
j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / a q u a - o n l i n e

Effects of temperature on reproduction and survival of the calanoid copepod

Pseudodiaptomus pelagicus
Andrew L. Rhyne a,⁎,2, Cortney L. Ohs a, Erik Stenn b,1
a
University of Florida, Indian River Research and Education Center, 2199 South Rock Road, Fort Pierce FL 34945, USA
b
AlgaGen LLC, 2336 20th Street, Vero Beach FL 32960, USA

a r t i c l e i n f o a b s t r a c t

Article history: Four experiments were conducted on the calanoid copepod, Pseudodiaptomus pelagicus, to determine the
Received 6 September 2008 effects of temperature (24, 26, 28, 30, 32, and 34 °C) on survival, development time, reproductive output, and
Received in revised form 18 March 2009 population growth in order to define the optimal temperature for culture. The first experiment stocked early
Accepted 21 March 2009
stage nauplii into 1 L beakers and cultured them using standard procedures until five days after the first
mature adults were observed; from this survival, sex ratio, time to maturation, and fecundity were measured.
Keywords:
Calanoid
The second and third experiments evaluated the effects of temperature on nauplii production by stocking
Copepod individual pairs and 25 pairs of adults, respectively; in both experiments nauplii production was determined
Pseudodiaptomus pelgicus daily for 10 days. The fourth experiment determined the effects of temperature on population growth and
Temperature composition of the population produced by stocking 10 adult pairs and culturing them for 10 days at six
temperatures. Results indicate survival from early nauplii to adult was significantly affected by temperature
and those cultured from 24–30 °C had the highest mean survival. Time to first maturation and maturation of
the entire population was significantly influenced by temperature and took from 6.8 to 12.8 days.
Temperature significantly affected nauplii production in both individual and groups of paired adults.
Temperature affected the mean daily nauplii production by decreasing the brood interval but did not affect
the mean brood size. The number of nauplii produced by 25 adult pairs was significantly influenced by
temperature; the optimal temperature was 27.5 °C at which 1861 nauplii were produced. The distribution of
developmental stages in the population was also affected by temperature; at lower temperatures the
population consisted of a greater proportion of nauplii while at 32 °C the population was comprised of more
advanced staged individuals. When developing production objectives, aquaculturists must consider
temperature because it has multiple effects on the culture of P. pelagicus. The optimal temperature range
to achieve high survival and the greatest nauplii production is 26–30 °C. To maintain long-term stock cultures
the best temperature may be 24 °C to slow maturation and growth while 28–32 °C may be used to maximize
nauplii production by decreasing time to maturation and decreasing brood intervals.
© 2009 Elsevier B.V. All rights reserved.

1. Introduction the addition of enrichments. Feeding copepods exclusively or in

combination with other live organisms has repeatedly demonstrated
In the wild, copepods are dominant prey items for the vast superior results in terms of growth, survival, and the overall health of
majority of marine fish larvae (Hunter, 1981; Leis, 1991; Østergaard larval fish (Kraul et al., 1992; 1993; Shields et al., 1999; Gardner, 2000;
et al., 2005; Sampey et al., 2007). Brachionus spp. (rotifers) or Artemia Payne et al., 2001; Støttrup, 2000; Støttrup, 2003; Wilcox et al., 2006).
spp. (Artemia) are commonly used in aquaculture because of Feeding copepods to larval fish species with small mouth gapes has
convenience and commercial availability (Hoff and Snell, 1999). Yet, allowed these species to be successfully cultured through the larval
Artemia nauplii and rotifers are deficient in essential highly unsatu- phase (Shields et al., 2003; Shields et al., 2005; Baensch, 2009).
rated fatty acids for many larval and juvenile marine fishes without Currently, our ability to commercially produce marine fish is generally
limited to those species that can be reared on enriched rotifers and
⁎ Corresponding author. Department of Biology and Marine Biology, Roger Williams Artemia.
University, One Old Ferry Road, Bristol, RI 02809, USA. Tel.: +1 401 254 5750; fax: +1 401 Despite the aforementioned advantages of feeding copepods
254 3310. over other live feeds currently in use, the use of copepods in
E-mail addresses: arhyne001@[Link], arhyne@[Link] (A.L. Rhyne), commercial settings is rare. This is primarily due to an inability to
cohs@ufl.edu (C.L. Ohs), eriks@[Link] (E. Stenn).
1
Tel.: +1 772 978 1395.
produce a reliable, continuous supply of copepods on a large-
2
Edgerton Research Laboratory, New England Aquarium, One Central Wharf, Boston, scale (Gapasin and Duray, 2000; Payne and Rippingale, 2001a;
MA 02110, USA. Støttrup, 2003). Typical culture densities for calanoid species are

0044-8486/$ – see front matter © 2009 Elsevier B.V. All rights reserved.
doi:10.1016/[Link].2009.03.041
54 A.L. Rhyne et al. / Aquaculture 292 (2009) 53–59

0.5–1.0/mL (Morehead, 2004), whereas rotifers have been reported to only adults. Early stage nauplii (N1–N3) were obtained by sieving
achieve a density of 16,000/mL in intensive recirculating systems stock cultures through a 125 µm nylon screen, and collecting the
(Suantika et al., 2001). nauplii on a 50 µm nylon screen.
Several species of copepods have shown commercial production
potential. Payne and Rippingale (2001a,b) demonstrated successful 2.2. Survival, sex ratio, maturation and fecundity
culture methods for Gladioferens imparipes, and designed a system to
produce 1–2 nauplii/mL on a continuous basis. G. imparipes is a small To determine the effects of temperature on survival, sex ratio,
egg bearing estuarine calanoid copepod with a semi-benthic adult life maturation, and fecundity, a series of experiments were conducted
stage (Payne and Rippingale, 2001a). Production of Acartia tonsa using early stage nauplii obtained from stock cultures at six
continues to be refined (Peck and Holste, 2006). Recent success with temperatures (24, 26, 28, 30, 32, and 34 °C). All other experimental
the long-term cold storage of A. tonsa eggs could prove to be a conditions were maintained per stock culture protocols. Each
valuable egg banking method (Drillet et al., 2006; Holmstrup et al., temperature treatment was replicated six times and cultures were
2006; Drillet et al., 2007). Several small paracalanid species have also maintained in a climate controlled room within constant temperature
recently shown promise for mass scale culture (McKinnon et al., 2003; water baths. Each replicate was a covered 1 L beaker which contained
Shields et al., 2005; Vanderlugt and Lenz, 2008). 650 mL of seawater. Saline water was obtained from the Atlantic
In 2003, we isolated the copepod, P. pelagicus, from the waters of Ocean (35–35.5 g/L) and filtered before use. A total of 200 nauplii
south Florida and have kept it in continuous culture for five years. were volumetrically stocked into each replicate beaker. Volumetric
P. pelagicus has exhibited culture characteristics very similar to G. stocking was conducted with a 10 mL pipette (Eppendorf Model
imparipes, and appears well suited for mass production. Pseudodiap- 022472208). To determine the accuracy we conducted counts on 12
tomus spp. are semi-benthic calanoid copepods; the adults are volumetric samples taken from a homogenized beaker of nauplii. In
substrate oriented and nauplii and early copepodites are pelagic these 12 volumetric samples we were able to collect 100 ± 6.5% of the
(Jacobs, 1961). The genus is globally distributed from tropical to desired number of nauplii.
temperate waters (Walter, 1989). Predominantly an estuarine genus, Each day, replicate beakers of copepods were observed to
they generally tolerate a wide range of environmental parameters determine the time to first maturity and time to total population
(Chen et al., 2006). Unlike many other calanoids, Pseudodiaptomus maturity. When P. pelagicus becomes sexually mature, adults
spp. appear well suited to culture systems because they can tolerate aggregate on the walls of the culture vessel and pair making it
heavy aeration, tolerate the presence of sediment and suspended relatively easy to determine the level of maturation. Females were
solids, grow and reproduce well on a single readily produced considered to be mature when they were carrying their first egg sac
microalgae species (Isochrysis galbana), and can achieve densities of (Payne and Rippingale, 2001b). First maturity was defined as when
over 5/mL (unpublished data). the first female was observed to be carrying eggs. The population was
Temperature is a key abiotic factor regulating the growth and determined to be totally mature when no free swimming copepodites
reproductive potential of copepods in marine systems (Santos et al., were observed and all females were gravid. Five days after first
1999; Peterson, 2001; Isla and Perissinotto, 2004; Sullivan et al., 2007; maturity, the cultures were sieved onto a 50 µm screen and then
Sun et al., 2008). Additionally, temperature is a key variable in the placed in 30 mL vials and preserved in a 5% solution of neutral
development of production regimes (Santos et al., 1999; Holste and buffered formalin in seawater and stored in a refrigerator (4 °C) until
Peck, 2006; Milione and Zeng, 2008). It is important to identify the enumeration. The entire population was counted, sexed and the
impacts of key abiotic factors prior to evaluating diet and other biotic number of ovigerous females was recorded. Survival was calculated by
culture conditions. For example, the microalga Rhodomonas lens, the total number of copepods harvested divided by the number of
commonly fed to copepods, is temperature sensitive and cultures copepods volumetrically stocked (200). Copepods were sexed by
decline or crash at temperatures above 26 °C. Therefore, this nutrient observing the morphologically distinct antennae, females exhibit
rich species of algae may not be feasible for feeding species of straight antennaes and males while the right antennule bent; they
copepods which require higher culture temperatures. also exhibit sexual size dimorphism, with the female being about 40%
A series of experiments were conducted to measure the effect of larger (Grice, 1969). Fecundity was determined by excising both egg
temperature on the survival, development time, reproductive output, sacs with fine forceps and needles from five females per replicate. Egg
and population growth of P. pelagicus with the overall objective to sac membranes were dissolved by placing each egg sac in a 5% solution
define the most suitable temperature or range of temperatures for of sodium hyperchlorite and gently agitating. Then the total number of
commercial production and future experimentation. eggs was quantified for each egg sac and each female with a stereo-
microscope (Olympus SZ30).
2. Materials and methods
2.3. Nauplii production
2.1. Stock cultures
To determine the effects of temperature on the timing and rate of
P. pelagicus stock cultures for this experiment were obtained from nauplii production, two separate experiments were conducted to
AlgaGen LLC located in Vero Beach, Florida and were of the strain evaluate both group and individual daily nauplii production at the six
PP1103. Established standard culture protocols were followed which treatment temperatures. The adult pairs of copepods were obtained
consisted of culture in 100 L static tanks at 26 °C and a salinity of 35 g/L. from stock cultures at each treatment temperature.
Gentle aeration was provided from the bottom of the tank. Water For group production, six replicate enclosures for each treatment
quality was maintained by exchanging 100% of the culture water were stocked with 25 reproductive adult pairs (male and female
every Monday and Thursday. Photoperiod was maintained at 24 h of attached) and cultured for 10 days. Enclosures were 1 L beakers
light. All copepods were provided a daily ration of Tahitian strain containing a 350 mL screen enclosure (165 µm nylon mesh on the
Isochrysis galbana (T-iso) from a stock culture to obtain a feeding bottom of the enclosure) nested inside of the beaker. Daily, the
density between 2 and 3 × 105 cells/mL. Unless otherwise stated, enclosures were removed from the beakers, retaining the adults on
experimental culture conditions were maintained during experi- the screen, and were immediately placed into a fresh beaker
ments in accordance with the standard protocols developed by containing temperature acclimated seawater and T-iso. Daily produc-
AlgaGen LLC described above. Adult copepods were obtained by tion was determined by counting the nauplii produced during each
sieving stock cultures through a 200 µm nylon screen which retained 24 h period.
 A.L. Rhyne et al. / Aquaculture 292 (2009) 53–59 55

Table 1
Mean (n = 6) survival, sex ratio, first maturity, population maturity, percent ovigerous, and fecundity of Pseudodiaptomus pelagicus cultured at the six treatment temperatures from
an initial population of 200 nauplii and cultured until five days after observation of the first ovigerous female.

Temperature Survival (%) Sex ratio First Population Ovigerous Fecundity

(°C) M:F maturity maturity (%) Left egg sac Right egg sac Total
(days) (days)
ab a a a a a a
24 81.3 ± 22.7 1.1 ± 0.2 10.7 ± 1.0 12.8 ± 0.4 50.5 ± 15.7 12.9 ± 1.2 3.2 ± 0.9 16.2 ± 1.5a
26 86.3 ± 24.6ab 1.0 ± 0.3a 9.3 ± 1.6ab 12.0 ± 0.0b 11.9 ± 6.4b 8.8 ± 1.0b 2.0 ± 0.9a 10.9 ± 6.4b
28 101.8 ± 9.0a 0.9 ± 0.2 a 7.8 ± 0.4bc 10.0 ± 0.0c 91.2 ± 6.3c 20.0 ± 1.2c 5.2 ± 0.8b 25.2 ± 1.0c
30 74.3 ± 9.8ab 1.0 ± 0.2 a 6.8 ± 0.4c 9.0 ± 0.0d 78.8 ± 9.0c 17.7 ± 1.9cd 6.0 ± 0.9b 23.7 ± 2.3cd
32 66.7 ± 8.9b 1.2 ± 0.1 a 6.7 ± 0.5c 8.0 ± 0.0e 66.7 ± 10.1c 15.3 ± 2.2ad 5.3 ± 1.0b 20.6 ± 2.6de
34 62.5 ± 10.0b 1.2 ± 0.2 a 6.7 ± 0.5c 8.0 ± 0.0e 53.0 ± 13.5a 14.0 ± 1.8a 5.0 ± 1.1b 19.1 ± 2.1ae

Different superscript letters indicate statistical differences (p ≤ 0.05) among treatments.

For individual production, reproductive adult pairs were individu- cultured between 24–30 °C had the higher mean survival than other
ally held in one of six 30 mL beakers per treatment, each containing temperatures and the highest survival of 101.8 ± 9.0% was recorded at
20 mL of temperature acclimated seawater and T-iso. Each day the 28 °C. Survival significantly declined in the 32 °C and 34 °C treatments
individual pair was captured with a transfer pipette and moved to a to 66.7 ± 8.9% and 62.5 ± 10.0%, respectively. Sex ratios of the final
new culture vessel containing fresh temperature adjusted seawater populations did not significantly (p N 0.05) differ from the expected
and T-iso. Daily nauplii production was determined by counting the 1:1 male:female ratio and the six treatments were not significantly
number of nauplii produced during each 24 h period. different from each other (p = 0.0869). Time to the first reproductive
For individual production, brood interval was the number of days female and to the total population maturity was strongly influenced
between broods, measured from the first day nauplii were present in by temperature. Development time from an early nauplii stage to the
two successive broods. A brood was defined as the production of first observed reproductive female was highly significant (p b 0.0001)
greater than one nauplii in the 24 h period between water exchanges. and ranged from 10.7 ± 1.0 days to 6.7 ± 0.5 days and followed a
Mean brood size was calculated from all broods during the 10 day decreasing trend with increasing temperature. Likewise, time to total
experimental culture period. population maturity was significantly influenced (p b 0.0001) by
temperature, taking 12.8 ± 0.4 days at 24 °C and 8.0 ± 0.0 days at
2.4. Population dynamics 34 °C. Fecundity was significantly affected (p b 0.0001) by rearing
temperature, ranging from 25.2 ± 1.0 eggs to 16.2 ± 1.5 eggs per
To determine the effects of temperature on the development of female, with the highest fecundity measured at 28 and 30 °C. In all
population composition, 10 reproductive adult pairs were obtained treatments, females always had a greater number of eggs in their left
from stock cultures and were stocked into six 1 L beakers containing egg sac compared to their right (p b 0.0001). At the time of sampling
650 mL of filtered seawater and T-iso at each of the six treatment the 28 and 30 °C treatments displayed a significantly higher
temperatures. After 10 days following standard culture protocols, the (p b 0.0001) percentage of ovigerous females (91.2 ± 6.3% and 78.8 ±
entire population was sieved onto a 50 µm screen then placed in 9.0%, respectively) than other treatments.
30 mL vials and preserved in a 5% neutral buffered formalin and
seawater solution and placed in a refrigerator (4 °C) until enumera- 3.2. Nauplii production
tion. The number of early nauplii, late nauplii (N4–N6), copepodites
(C1–C5), and adults (C6) (Grice, 1969) was quantified for each Temperature significantly affected nauplii production in both
replicate by placing the population on a zooplankton counting wheel individual pairs (p = 0.0012) and in groups of pairs (p b 0.0001),
and observing each individual with a stereo-microscope. however, the mean brood size was not affected by temperature
(p = 0.8991) (Table 2). Temperature affected the mean daily nauplii
2.5. Statistics production by decreasing the amount of time required between
broods (brood interval) as temperature increased. The brood interval
An analysis of variance using the general linear model (PROC GLM) decreased from 1.9 ± 0.5 days at 24 °C to 1.3 ± 0.1 days at 32 °C. Water
of SAS (SAS, 1999) was used to determine if there were statistically temperature of 34 °C impeded reproductive function and an analysis
significant differences between treatments for survival, time to first was not possible due to the low number of broods. In groups, a
maturity, time to population maturity, percent ovigerous, fecundity, significant decrease (p = 0.0014) occurred in the total number of
brood interval, brood size, and total nauplii produced. The means were nauplii produced, once temperatures reached 32 °C (Fig. 1). For
separated by the Tukey's procedure of SAS (SAS, 1999). Statistical individual pairs, a significant decrease (p b 0.0001) in total nauplii
significance occurred in all analyses when the calculated p-value was produced only occurred at 34 °C (Table 2).
≤0.05. All mean values are reported as mean ±S.D.
A chi square analysis was conducted on the male:female sex ratio,
and when comparing the number of eggs in the left and right egg sacs,
to determine if the results were significantly different from the Table 2
Mean (n = 6) brood interval, size, and total nauplii production for Pseudodiaptomus
expected 1:1 ratio (SAS, 1999). The regression curves and formulas pelagicus cultured in individual pairs at the six treatment temperatures.
were generated with Sigma Plot Version 8.0 software (Sigma Plot,
2002). Temperature Brood interval Brood size Total nauplii production
(°C) (Days)
24 1.9 ± 0.5a 16.0 ± 4.0a 87.8 ± 10.7a
3. Results
26 1.6 ± 0.2ab 14.5 ± 4.0a 80.8 ± 17.3a
28 1.4 ± 0.1ab 15.1 ± 3.9a 98.7 ± 25.8a
3.1. Survival, sex ratio, maturation and fecundity 30 1.4 ± 0.2b 16.2 ± 3.7a 90.3 ± 36.5a
32 1.3 ± 0.1b 15.1 ± 3.0a 86.7 ± 37.2a
Survival of P. pelagicus from early nauplii to adult was significantly 34 – 14.8 ± 2.9a 21.8 ± 21.7b

(p = 0.004) affected by culture temperature (Table 1). Copepods Different superscript letters indicate statistical differences (p ≤ 0.05) among treatments.
56 A.L. Rhyne et al. / Aquaculture 292 (2009) 53–59

(p b 0.0001) (Figs. 3 and 4). The total population was similar from
24–30 °C and peaked at 30 °C with 386.8 ± 186.5 individuals, followed
by a very large decline at 34 °C to 13.0 ± 14.1 individuals (Fig. 4).
The distribution of developmental stages within the population
was also affected by temperature (Fig. 3). At lower temperatures, the
population had a larger number of nauplii and copepodites than at
higher temperatures up to 32 °C; the 34 °C treatment performed
poorly. The number of adults in the population reached a maximum at
30 °C before declining sharply at 32 and 34 °C. Despite the overall
decline in numbers, the 32 °C treatment was comprised of more
advanced staged individuals than nauplii. The presence of gravid
females increased with increasing temperatures and peaked at 32 °C.
At 34 °C, the population declined greatly in number, and the
distribution of life stages was no longer relevant.

4. Discussion

Temperature is often the most important environmental factor

Fig. 1. Total daily nauplii (Pseudodiaptomus pelagicus) produced at the six treatment affecting the productivity of copepods in natural systems (Christou
temperatures during 10 day group experiment (n = 6).
and Moraitou-Apostolopoulou, 1995; Siokou-Frangou, 1996). In our
study of P. pelagicus, temperature affected survival, maturation time,
the number of ovigerous females, and fecundity, but had no effect on
In the group experiment, daily nauplii production was much sex ratio. Sex ratio did not deviate from the expected 1:1 in all
higher for day one when compared to all other days regardless of treatments, which is consistent with the calanoid copepod, G.
temperature (Fig. 2). This sharp decrease in production was followed imparipes (Rippingale and Hodgkin, 1974). Survival and fecundity
by an increase in production for days three to six. Total nauplii was highest at 28 °C, and survival was lowest at 32 and 34 °C.
production in the group experiment increased to 1861 nauplii at Fecundity was lowest at 24 and 26 °C. The percent ovigerous females
27.5 °C, then declined gradually until 30 °C before sharply decreasing followed the same trend as fecundity and was lowest at 26 °C. This
(Fig. 1). may be a result of a longer interbrood duration; additionally, the time
Regardless of scale, individual pairs, or groups of pairs, the trend of when the population was sampled may have been a time when the
temperature on mean daily nauplii production was similar. Mean daily majority of females were between broods. Mean development time,
nauplii production in the individual pairs and group experiments from early nauplii to reproductive adults, decreased exponentially
were both highest at 28 and 30 °C. Daily production peaked at 28 °C in with increasing temperature and reached the shortest duration at
the individual pair experiments, and had 9.8 ± 2.6 nauplii per female 32 °C. An exponential increase in development time with a
per day while the group experiment also peaked at 28 °C with corresponding decrease in temperature is well supported in the
production of 9.5 ± 1.9 nauplii per female per day. literature. The calanoid copepod, Pseudocalanus newmani, was
reported to experience a doubling of development time from 20.9 to
3.3. Population dynamics 42.3 days when temperatures decreased from 15 to 6 °C (Lee et al.,
2003). This trend was also observed in A. clausi, where development
The effects of temperature on the population composition and the time increased from 35.4 to 74.8 days when temperatures decrease
total number in the population were both highly significant from 10 to 5 °C. This effect is also well documented in marine

Fig. 2. Mean (n = 6) daily production of Pseudodiaptomus pelagicus nauplii cultured at the six treatment temperatures in groups of pairs following 10 days of culture.
 A.L. Rhyne et al. / Aquaculture 292 (2009) 53–59 57

Fig. 3. Mean (n = 6) number of five life stages, early nauplii (N1–N3), late nauplii (N4–N6), copepodites (C1–C5), adults (C6), and gravid females within the population of
Pseudodiaptomus pelagicus cultured for 10 days at the six treatment temperatures from an initial population of 10 adult pairs.

harpacticoid copepods (Williams and Jones, 1999). As temperatures Population growth and composition has recently been used to
increase to the upper thermal limit of the species, the effect of examine the effects of temperature and salinity on the aquaculture
temperature on development time decreases in magnitude (see production of A. singiensis (Milione and Zeng, 2008). Milione and Zeng
Peterson, 2001 for a review). (2008) observed the highest production between 25–30 °C with a
In our study of P. pelagicus development, the regression curve peak at 30 °C followed by a sharp decline at 34 °C. Furthermore, they
became asymptotic at approximately 30 °C. Temperatures greater than related this result to the optimal temperature where mean develop-
30 °C were detrimental to survival, percent ovigerous females, and ment time was shortest and survival and egg production was highest.
fecundity. Culture temperatures below 28 °C had high survival and This is evident in our results in which the optimal observed range for
experienced lower fecundity and extended maturation time from production of P. pelagicus was 26–30 °C, with the highest at the 28 °C
early nauplii to adult. The optimal temperature for aquaculture treatment and the optimal temperature of 27.5 °C predicted by the
purposes appears to be 30 °C, which results in the shortest duration quadratic function. Temperatures above 30 °C elicit the sharp decline
time to adult and relatively high survivorship and fecundity. observed in Fig. 4. In addition to an increased total production, the
Nauplii production from both individual pairs and groups of pairs composition of the population is directly affected by temperature. A
followed similar trends with peak production at approximately 28 °C. larger proportion of the population reached maturity at 28 °C which
Individual pair data revealed that increased production was not due to an resulted in peak performance. Above 30 °C, culture performance
increase in brood size but rather a decrease in brood interval. Brood declines and stage composition reflects the thermal stress effects.
interval followed the same trend as mean development time, and Population growth and composition is a good indicator of potential
exponentially increased with decreasing temperature. Brood interval at aquaculture performance because it shows the effects of temperature
28 °C was 1.4 days and at 34 °C thermal stress likely impeded on reproduction, growth, and survival. However, this method does not
reproduction. At 34 °C, production was lower than all other temperatures provide necessary data to determine the effects of temperature on
and a brood interval could not be determined, although mean brood size
was similar to other temperatures. At 34 °C, thermal stress resulted in
more erratic production of nauplii between replicates with a range of 0–
55 nauplii produced in a 10 day period. Also, a greatly reduced lifespan
was observed at 34 °C and the maximum number of observed broods was
three in one replicate but production ceased after the fourth day. This may
indicate thermal stress caused energy to be allocated toward survival
processes and away from reproduction. This trend was reported in Tisbe
battagliai, where at 25 °C nauplii production ceased after 20 days while
lower temperature treatments continued to produce nauplii for 36 days
(Williams and Jones, 1999). Group data confirmed this with a similar
pattern of production. Daily mean nauplii production was similar in the
grouped and individual pairs, and the trend remained the same. The
elevated production in the group experiment on the first day suggests a
possible container effect and/or possible stress of the copepods having to
acclimate to experimental conditions. Despite the apparent confounding
effect of the enclosure, the overall pattern remains constant in all
experiments, increasing temperature increases production up to
30 °C, after which as temperature increases production declines. The
overall trend in the data corroborates that of the individual pairs Fig. 4. Total population of Pseudodiaptomus pelagicus produced at the six treatment
where optimal production occurred at 26–30 °C. temperatures during the population dynamics experiment (n = 6).
58 A.L. Rhyne et al. / Aquaculture 292 (2009) 53–59

mean brood size or brood interval. Individual pair data showed the Grice, G.D., 1969. The developmental stages of Pseudodiaptomus coronatus Williams
(Copepoda, Calanoida). Crustaceana 17, 291–301.
mean brood size was constant and the decrease in production at lower Hoff, F.H., Snell, T.W., 1999. Plankton Culture Manual, fifth edition. Florida Aqua Farms
temperatures was due to the longer brood interval while at higher Inc., Dade City, FL. 160 pp.
temperatures it was from decreases in survival and number of broods. Holste, L., Peck, M.A., 2006. The effects of temperature and salinity on egg production
and hatching success of Baltic Acartia tonsa (Copepoda: Calanoida): a laboratory
Experimental scale is an issue in copepod production studies. Few investigation. Marine Biology 148, 1061–1070.
studies have examined production at a large commercial scale. Payne and Holmstrup, M., Overgaard, J., Sørensen, T.F., Drillet, G., Hansen, B.W., Ramløv, H., Engell-
Rippingale (2001a) examined three different production systems ranging Sørensen, K., 2006. Influence of storage conditions on viability of quiescent copepod
eggs (Acartia tonsa Dana): effects of temperature, salinity and anoxia. Aquaculture
from 60 to 1000 L with the peak nauplii production of 1117/L/day in 60 L
Research 37, 625–631.
batch systems compared to 520/L/day in 1000 L semi-continuous Hunter, J., 1981. Feeding ecology and predation of marine fish larvae. In: Lasker,, R. (Ed.),
systems. It is possible that nauplii production may change when scale is Marine Fish Larvae: Morphology, Ecology, and Relation to Fisheries. University of
Washington Press, Seattle, WA, pp. 33–77.
increased. However, conducting replicated experiments at commercial
Isla, J.A., Perissinotto, R., 2004. Effects of temperature, salinity and sex on the basal
scale can be impractical given the cost and time involved. Small scale metabolic rate of the estuarine copepod Pseudodiaptomus hessei. Journal of
studies, such as the present, provide valuable data concerning abiotic Plankton Research 26, 579–583.
parameters which remain constant regardless of scale. Jacobs, J., 1961. Laboratory cultivation of the marine copepod Pseudodiaptomus
coronatus Williams. Limnology and Oceanography 6, 443–446.
Kraul, S., Brittain, K., Cantrell, R., Nagao, T., Ako, H., Ogasawara, A., Kitagawa, H., 1993.
5. Conclusion Nutritional factors affecting stress resistance in the larval mahimahi, Coryphaena
hippurus. Journal of the World Aquaculture Society 24, 186–193.
Kraul, S., Nelson, A., Brittain, K., Ako, H., Ogasawara, A., 1992. Evaluation of live feeds for larval
Temperature significantly affected the growth, survival, and and postlarval mahimahi Coryphaena hippurus. Journal of the World Aquaculture Society
reproductive output of the semi-benthic calanoid copepod P. pelagicus 23, 299–307.
as reported for other species of calanoid (Peterson, 2001) and Lee, H., Ban, S., Ikeda, T., Matsuishi, T., 2003. Effect of temperature on development,
growth and reproduction in the marine copepod Pseudocalanus newmani at
harpacticoid copepods (Williams and Jones, 1999). The optimal satiating food condition. Journal of Plankton Research 25, 261–271.
range for production was observed to be 28–30 °C; this range provides Leis, J.M., 1991. The pelagic stage of reef fishes. In: Sale, P.F. (Ed.), The Ecology of Fishes
for high survival, short brood interval, and decreased mean develop- on Coral Reefs. Academic Press, San Diego, CA, pp. 183–230.
Morehead, D., 2004. Copepod culture workshop in Hawaii. Austasia Aquaculture 18,
ment time. Temperatures below the optimal range resulted in slower 46–49.
growth and longer brood intervals, which contributed to overall lower McKinnon, A.D., Duggan, S., Nichols, P.D., Rimmer, M.A., Semmens, G., Robino, B., 2003.
production. Additionally, temperatures above the optimal range The potential of tropical paracalanid copepods as live feeds in aquaculture.
Aquaculture 223, 89–106.
resulted in a marked decline in production as a result of reduced
Milione, M., Zeng, C., 2008. The effects of temperature and salinity on population
survival and a decreased number of broods. Mean sex ratio and brood growth and egg hatching success of the tropical calanoid copepod, Acartia sinjiensis.
size were not affected by temperature. Aquaculture 275, 116–123.
Based upon production goals, these results provide options for Østergaard, P., Munk, P., Janekarn, V., 2005. Contrasting feeding patterns among species
of fish larvae from the tropical Andaman Sea. Marine Biology 146, 595–606.
commercial production. Culture water temperatures of 28–30 °C will Payne, M.F., Rippingale, R.J., 2001a. Intensive cultivation of the calanoid copepod
maximize population growth and nauplii production. Temperatures from Gladioferens imparipes. Aquaculture 201, 329–342.
20–24 °C will slow development and nauplii production but can be used Payne, M.F., Rippingale, R.J., 2001b. Effects of salinity, cold storage and enrichment on
the calanoid copepod Gladioferens imparipes. Aquaculture 201, 251–262.
to maintain cultures long-term with high survival while decreasing algae Payne, M.F., Rippingale, R.J., Cleary, J.J., 2001. Cultured copepods as food for west
requirements. Manipulating temperature within a production scenario Australian dhufish (Glaucosoma hebraicum) and pink snapper (Pagrus auratus)
with multiple culture systems facilitates the timing of maturation and larvae. Aquaculture 194, 137–150.
Peck, M.A., Holste, L., 2006. Effects of salinity, photoperiod and adult stocking density on
nauplii production by controlling developmental time and provides the egg production and egg hatching success in Acartia tonsa (Calanoida: Copepoda):
ability to coincide or stagger the timing of nauplii production. optimizing intensive cultures. Aquaculture 255, 341–350.
Peterson, W.T., 2001. Patterns in stage duration and development among marine and
freshwater calanoid and cyclopoid copepods: a review of rules, physiological
Acknowledgements constraints and evolutionary significance. Hydrobiologia 453/454, 91–105.
Rippingale, R.J., Hodgkin, E.P., 1974. Population growth of a copepod Gladioferens imparipes
We thank the Florida Department of Agriculture and Consumer Thomson. Australian Journal of Marine and Freshwater Research 25, 351–360.
Sampey, A., McKinnon, A.D., Meekan, M.G., McCormick, M.I., 2007. Glimpse into guts:
Services Division of Aquaculture and the University of Florida Institute
overview of the feeding of larvae of tropical shorefishes. Marine Ecology Progress
of Food and Agricultural Sciences (IFAS) Research Innovation Fund Series 339, 243–257.
Grants Program for funding this research. We thank the three Santos, P.J.P., Castel, J., Souza-Santos, L.P., 1999. Development time of harpacticoid
anonymous reviewers for valuable comments. We also thank Scott copepods: some empirical models and implications. Journal of the Marine
Biological Association of the U.K. 79, 1123–1124.
Grabe, Matthew DiMaggio, and Shawn DeSantis for assisting with SAS, 1999. SAS Procedures Guide, Version 8. SAS Publishing. 1624 pp.
various aspects of these experiments. Shields, R.J., Bell, J.G., Luizi, F.S., Gara, B., Bromage, N.R., Sargent, J.R., 1999. Natural
copepods are superior to enriched Artemia nauplii for halibut larvae (Hippoglossus
hippoglossus) in terms of survival, pigmentation and retinal morphology: relation
References to dietary essential fatty acids. Journal of Nutrition 129, 1186–1194.
Shields, R.J., Kotani, T., Augustin, M., Marion, K., Kobashigawa, J., 2003. Intensive culture
Baensch, F., Tamaru, C., 2009. Spawning and development of larvae and juveniles of the of a calanoid copepod, Parvocalanus sp., and application for rearing small sub-
rare blue Mauritius angelfish, Centropyge debelius (1988), in the hatchery. Journal tropical marine fish larvae. World Aquaculture Annual Conference, Salvador Brazil.
of the World Aquaculture Society 40 (4). Book of Abstracts, vol. 2. 720 pp.
Chen, Q., Sheng, J., Lin, Q., Gao, Y., Lv, J., 2006. Effect of salinity on reproduction and survival Shields, R.J., Kotani, T., Molnar, A., Marion, K., Kobashigawa, J., Tang, L., 2005. Intensive
of the copepod Pseudodiaptomus annandalei Sewell, 1919. Aquaculture 258, 575–582. cultivation of a subtropical paracalanid copepod, Parvocalanus sp., as a prey item for
Christou, E.D., Moraitou-Apostolopoulou, M., 1995. Metabolism and feeding of small marine fish larvae. In: Lee, C.S., O'Bryan, P.J., Marcus, N. (Eds.), Copepods in
mesozooplankton of the eastern Mediterranean coast (Hellenic waters). Marine Aquaculture. Blackwell Publishing, pp. 209–223.
Ecology Progress Series 126, 39–48. Sigma Plot, 2002. Sigma Plot Users Guide, Version 8.0. SPSS Inc. 526 pp.
Drillet, G., Iversen, M.H., Sørensen, T.F., Ramløv, H., Lund, T., Hansen, B.W., 2006. Effect of Siokou-Frangou, I., 1996. Zooplankton annual cycle in a Mediterranean coastal area.
cold storage upon eggs of a calanoid copepod Acartia tonsa Dana and their Journal of Plankton Research 18, 203–223.
offspring. Aquaculture 254, 714–729. Støttrup, J.G., 2000. The elusive copepods: their production and suitability in marine
Drillet, G., Lindley, L.C., Michels, A., Wilcox, J., Marcus, N.H., 2007. Improving cold aquaculture. Aquaculture Research 31, 703–711.
storage of subitaneous eggs of the copepod Acartia tonsa Dana from the Gulf of Støttrup, J.G., 2003. Production and nutritional value of copepods. In: Støttrup, J.G., McEvoy,
Mexico (Florida — USA). Aquaculture Research 38, 457–466. L.A. (Eds.), Live Feeds in Marine Aquaculture. Blackwell Science, Oxford, UK. 318 pp.
Gapasin, R.S.J., Duray, M.N., 2000. Effects of DHA enriched live food on growth and Suantika, G., Dhert, P., Rombaut, G., Vanderberghe, J., DeWolf, T., Sorgeloos, P., 2001. The use
survival and incidence of opercular deformities in milkfish (Chanos chanos). of ozone in a high density recirculation system for rotifers. Aquaculture 201, 35–49.
Aquaculture 193, 49–63. Sullivan, B.K., Costello, J.H., Van Keuren, D., 2007. Seasonality of the copepods Acartia
Gardner, T., 2000. Spawning and rearing the yellow dottyback, Pseudochromis fuscus. hudsonica and Acartia tonsa in Narragansett Bay, RI, USA during a period of climate
Freshwater and Marine Aquarium 23 (4), 126–132. change. Estuarine, Coastal and Shelf Science 73, 259–267.
 A.L. Rhyne et al. / Aquaculture 292 (2009) 53–59 59

Sun, X.H., Sun, S., Li, C.L., Zhang, G.T., 2008. The seasonal and spatial variation in Wilcox, J.A., Tracy, P.L., Marcus, N.M., 2006. Improving live feeds: effect of a mixed diet of
abundance and egg production of Paracalanus parvus (Copepoda: Calanoida) in/ copepod nauplii (Acartia tonsa) and rotifers on the survival and growth of first-
out Jiaozhou Bay, China. Estuarine, Coastal and Shelf Science 79, 637–643. feeding larvae of the southern flounder, Paralicththys lethostigma. Journal of the
Vanderlugt, K., Lenz, P.H., 2008. Management of nauplius production in the paracalanid, World Aquaculture Society 37, 113–120.
Bestiolina similes (Crustacea: Copepoda): effects of stocking densities and culture Williams, T.D., Jones, M.B., 1999. Effects of temperature and food quality of the
dilution. Aquaculture 276, 69–77. reproduction of Tisbe battagliai (Copepoda: Harpacticoida). Journal of Experimental
Walter, T.C., 1989. Review of the New World species of Pseudodiaptomus (Copepoda: Marine Biology and Ecology 236, 273–290.
Calanoida), with a key to the species. Bulletin of Marine Science 45, 590–628.