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APPLIED RESEARCH
Advancements in hatchery production of red
snapper Lutjanus campechanus: Exclusive use of
small strain rotifers as initial prey for larval rearing
Yole Buchalla
1
| Charles James McGuigan
1
|
John D. Stieglitz
1
| Ronald H. Hoenig
1
| Carlos E. Tudela
1
|
Kimberly G. Darville
1
| Leonardo Ibarra-Castro
2
| Daniel Benetti
1
1
Department of Marine Biology and Ecology,
University of Miami—Rosenstiel School of
Marine, Atmospheric, and Earth Science,
Miami, Florida, USA
2
School of Forest, Fisheries & Geomatics
Sciences, University of Florida—Whitney
Laboratory for Marine Bioscience, St
Augustine, Florida, USA
Correspondence
Yole Buchalla, Department of Marine Biology
and Ecology, University of Miami- Rosenstiel
School of Marine, Atmospheric, and Earth
Science, 4600 Rickenbacker Causeway,
Miami, FL 33149, USA.
Email: ybuchalla@iattc.org
Present address
Yole Buchalla, Inter-American Tropical Tuna
Commission, 8901 La Jolla Shores Drive, La
Jolla, California 92037, USA.
Funding information
National Oceanic and Atmospheric
Administration and Florida Sea Grant,
Grant/Award Number: NOAA-OAR-SG-2017-
2005176; NOAA Sea Grant Award,
Grant/Award Number: NA17OAR4170318;
Florida Sea Grant, Grant/Award Number:
R/LR-A-58
Abstract
The red snapper (Lutjanus campechanus) has one of the
most valued fisheries in the southeastern United States and
has been largely studied for its aquaculture potential with
varied results that revealed production challenges. This
study focused on addressing challenges in larval rearing and
juvenile production of this species. Broodstock fish were
acclimated in a 60 m
3
tank equipped with recirculating
aquaculture system (RAS) and temperature control. After
1 year in captivity, fish volitionally spawned and larval rea-
ring trials were conducted in replicated flow-through sea-
water tanks ranging from 0.4 to 2.4 m
3
, at a temperature
range of 24–26C. Enriched s-strain rotifers, Brachionus
rotundiformis (lorica length of 100–210 μm), were used for
first feeding. Enriched Artemia sp. were gradually intro-
duced after 18 days post hatch (DPH). The survival rate
before metamorphosis (12 DPH) averaged 66.09 ± 0.08%.
Survival rate from yolk-sac larvae to post-metamorphic early
juvenile averaged 4.43 ± 0.01%. At 40 DPH, early red
snapper juveniles averaged 0.35 ± 0.02 g in weight and
28.83 ± 0.60 mm in standard length. A total of 31,849 fully
Received: 25 February 2022 Revised: 3 April 2023 Accepted: 10 May 2023
DOI: 10.1111/jwas.12992
This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which
permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no
modifications or adaptations are made.
© 2023 The Authors. Journal of the World Aquaculture Society published by Wiley Periodicals LLC on behalf of World Aquaculture
Society.
J World Aquac Soc. 2023;1–14. wileyonlinelibrary.com/journal/jwas 1
weaned juveniles were harvested. This study demonstrates
that small strain rotifers can be used exclusively as first feed-
ing prey organism for red snapper larvae, representing a sig-
nificant advancement for red snapper juvenile production.
KEYWORDS
aquaculture, broodstock, first feeding, larval husbandry, marine,
rotifers
1|INTRODUCTION
The red snapper Lutjanus campechanus is one of the most economically important marine fish species in the Gulf of
Mexico and South Atlantic coast of United States (Bardon-Albaret & Saillant, 2017). Overfishing led to intensive
management of the red snapper stock since 1990 (Bardon-Albaret et al., 2013); and since 2018 the species is no lon-
ger considered overfished (NOAA, 2020). Nonetheless, fishery closures remain in place throughout most of their
range. The popularity of red snapper as a gamefish, its high market value, and its importance to commercial fisheries
suggests that the development of husbandry techniques for this species could potentially lead to the development
of its aquaculture industry, as well as provide a source of fry for stock enhancement programs (Bardon-
Albaret, 2014; Bardon-Albaret & Saillant, 2017; Chávez et al., 2014; Ogle & Lotz, 2006; Watanabe et al., 2005).
Previous research conducted on red snapper focused on describing the early larval stages as well as spawning
induction techniques for wild-caught broodstock (Arnold et al., 1978; Bardon-Albaret et al., 2013; Hastey
et al., 2013; Papanikos et al., 2008; Rabalais et al., 1980; Saillant et al., 2013; Williams et al., 2004). While providing
the baseline for hatchery technology development for this species, much of this work was constrained by difficulties
in obtaining volitional spawning in captivity and high mortalities during the early developmental stages associated
with first feeding of the larvae. Similar to other Lutjanids, red snapper larvae experience rapid yolk-sac absorption
coupled with small mouth gape, creating a delicate phase during the switch from endogenous reserves to exogenous
feeding (Estrada-Godínez et al., 2015; Gutiérrez-Sigeros et al., 2018).
Several authors have reported that the success of red snapper larval rearing was dependent upon the use of cope-
pods as a first-feeding prey item due to their small sizes (50–180-μlength) and high natural nutritional value (Ogle
et al., 2005; Ogle & Lotz, 2006; Phelps et al., 2005; Rhodes & Phelps, 2008; Watanabe et al., 2005). Calanoid copepods
(Acartia,Calanus, and others), harpacticoid (Euterpina,Tisbe) and cyclopoid (Corycaeus,Oithona, and others) were success-
fully used during the first feeding of several snapper species including: golden snapper (Lutjanus johnii), mangrove red
snapper (Lutjanus argentimaculatus), spotted rose snapper (Lutjanus guttatus) (Alvarez-Lajonchère et al., 2012;Doi
et al., 1997;Schipp,2006), as well as in ornamental marine fish species with small mouth gapes (DiMaggio, 2020;Lee
et al., 2018). However, the scale of copepod culture necessary to support mass aquaculture production would likely be
cost-prohibitive and require extensive space in a commercial setting (Stottrup, 2000), suggesting that live prey alternatives
are necessary to support the development of commercial aquaculture of marine fish species such as the red snapper.
Snapper species have mouth width of approximately 190–220 μm at first feeding, and L. campechanus has a
mandibular length of 221 μm at 60 h post fertilization (Castaños, 1997; Williams et al., 2004; Zavala-Leal
et al., 2015). Small strain rotifers (s-strain rotifer), Brachionus rotundiformis, have a lorica length of 100–210 μm,
which is slightly over 60% of the relative predator's mouth width. Studies on prey selectivity of marine fish larvae
have repeatedly reported that larvae consume prey sizes that are 20–80% of their mouth width during first feeding
(Castaños, 1997; Saillant et al., 2013), which indicate that the size of s-strain rotifers could be adequate for first feed-
ing of snappers. It has also been suggested that the lack of successful attempts to feed red snapper larvae with roti-
fers could be related to inadequate enrichment rather than to prey size (Saillant et al., 2013).
2BUCHALLA ET AL.
Moreover, the size threshold of prey ingested by the larvae may not only be a function of larvae length or mouth
width but can also be affected by the density of available food (Duray & Kohno, 1990). Castaños (1997) have found
s-strain rotifers ranging 62–166 μm in the gut of L. argentimaculatus during the first days of feeding in an experimen-
tal larval rearing, potentially indicating that the excess rotifers in the tank likely grew, reproduced, and the neonates
were ingested by the larvae. Physical processes operating on a microscale level can also affect the first feeding of
marine fish larvae (Margulies et al., 2001). Hence, there are more factors other than prey size that can determine the
first-feeding success of marine fish larvae.
Along with copepods, enriched rotifers have been used effectively as first feeding organisms for snapper species
(Duray et al., 1996; Ibarra-Castro et al., 2020), small mouth-gape marine fish larvae (Alvarez-Lajonchère et al., 1996;
Geng et al., 2019; Wullur et al., 2011), and even challenging species that also exhibit rapid yolk-sac absorption and
high nutritional demand, such as scombrid larvae (Blanco et al., 2020; Kaji et al., 1999; Sawada et al., 2005; Stein
et al., 2018). S-strain rotifers are relatively easy to culture in large scale, compared to copepod species (Saillant
et al., 2013). Therefore, demonstrating the viability of using enriched rotifers during the initial feeding can provide a
viable alternative source of live prey and facilitate the development of mass production of L. campechanus, and other
marine species known to be difficult to raise.
In 2018 the University of Miami Experimental Hatchery (UMEH) began conducting research with red snapper
aimed at developing hatchery technology of economically important reef fish species of the Gulf of Mexico and
Caribbean. The objective of the present study was to test if the use of enriched s-strain rotifers (B. rotundiformis)
could be adequate as first feeding prey and enable success in the scaling-up of hatchery production for this species.
Data and results obtained on broodstock volitional spawning, larval rearing, and juvenile production of red snapper
at 40 days post hatch (DPH) are reported.
2|MATERIALS AND METHODS
2.1 |Egg production
A group of thirteen (13) wild fish (seven females averaging 6.14 ± 0.33 kg and six males averaging 5.30 ± 0.25 kg in
body weight after acclimation) was captured using hook-and-line techniques off the Atlantic coast of Florida. The
broodstock fish were conditioned over a period of 1 year in a maturation system previously described by Benetti
et al. (2008) and Stieglitz et al. (2012,2017).
Salinity was consistent at 36-ppt, and pH was 8.1. A shade-cloth cover was placed approximately two meters
above the tank to attenuate light intensity, whereas photoperiod was natural (25440200N, 80904300 W).
Broodstock red snapper were fed with a diet of previously-frozen cut fish—sardines (Sardina sp) and mackerel
(Scomber sp)–and market squid (Loligo sp), given six times a week to 2–3% of the total biomass. As a complementary
diet, a semi-moist feed (MADMAC-MS; Aquafauna Bio-Marine Inc. Hawthorne, CA) with the addition of a vitamin
and mineral mix was given to the fish once a week. The broodstock diet was supplemented with shrimp and poly-
chaetes (ProChaete; Sea Farms Nutrition, Miami, FL) in the 2 months preceding the spawning season.
The broodstock were conditioned to spawn volitionally through temperature control. Temperature regimes were
set to follow natural temperature fluctuations in the ocean in the fall and summer in the southeastern United States.
The first natural spawns were obtained when temperatures reached 25.8C, and subsequent spawns were observed
regularly while temperature was maintained 25.9 ± 0.04C. Fish spawned during the night and eggs were collected
in the morning from a 0.2 m
3
collector tank with a 0.2-mm nylon mesh screen when embryos were at the early
development stages. To determine the number of eggs/ml, three samples of 1-ml were counted from the spawns.
The mean ± standard error of the mean (SEM) of 1932 ± 63 eggs/ml was obtained. The total number of eggs were
estimated volumetrically using a 2-L graduated beaker and the fertilization rate was estimated by the relation
between the total number of floating eggs with the total number of eggs collected (floating and sinking).
BUCHALLA ET AL.3
Egg measurements were obtained from sub-samples of 10 eggs in 10 different spawns with the use of an optical
microscope (Leica Microscopes CME; Buffalo, NY).
Hatch rates were determined by volumetric sampling and visual counts of yolk-sac larvae 30 h post hatch (HPH)
taken directly from the incubation tanks. The averages were estimated from three samples, and the values are
expressed as mean ± SEM.
2.2 |Larval rearing
Two larval rearing trials were conducted using s-strain rotifers as first-feeding prey item in an experimental system con-
sisting of eight replicated 0.4 m
3
tanks, and in a pilot-scale system of four replicated 2.4 m
3
tanks with flow-through sea-
water. The cylindrical fiberglass tanks had dark green/blue walls and light green/blue bottoms. Seawater was filtered by
two sand filters filled with broken glass filter medium (Vitroclean; Trivitro Corporation, Kent, WA), followed by a series
of cartridge filters (10, 5, 1, and 0.05 μm) and a 200 W UV sterilization filter (Aqualogic; Inc, San Diego, CA).
Egg collection and treatment were described in detail by Benetti et al. (2008) and Stieglitz et al. (2012,2017).
Approximately 30 min after collection, the eggs were stocked in one 0.4 m
3
and one 2.4 m
3
tank and treated with
100-ppm Formalin (Parasite-S; Syndel Laboratories, Ferndale, WA) for 1 h. After that, water exchange was set to
1000% of the volume of the tank per day to flush the Formalin. Medium to strong aeration was supplied by an air
pump through an air ring placed around the bottom of the center standpipe of the incubation tanks and pure oxygen
was also provided to maintain saturation levels between 7 and 9 mg/L. Eggs were incubated at a mean density of
351 ± 13.96 eggs/L at a temperature of 25.5 ± 1C.
After the eggs hatched and the number of yolk-sac larvae was estimated, larvae (1 DPH) were evenly transferred
via beaker between the eight 0.4 m
3
tanks at a density of 35 larvae/L and to the four 2.4 m
3
tanks at a density of
73.08 ± 9.73 larvae/L. Each larval culture tank was fitted with a 200-μmesh screen in the center standpipe to allow
water exchange. During larval rearing, the mesh size was increased to 300 μafter 15 DPH, and 500 μafter 25 DPH.
Water exchange and aeration was gradually increased as larvae developed. During the incubation period, the
water flow was set to 1000% of the volume of the tank per day, and aeration was maintained strong enough to avoid
agglomeration of the eggs on the surface. Once the eggs hatched, water flow was set to 250–300% of the volume
of the tank per day and a gentle aeration was provided to avoid any mechanical shock and stress to the yolk-sac lar-
vae. Water exchange was maintained at the same rate for the first 6 days and it was gradually increased throughout
the run, as the biological load increased in the culture tanks and the larvae became more resilient. After 6 DPH,
water exchange was set to 400–500% of the volume to the tank per day until 14 DPH; then increasing to
500–600% from 14 DPH to 24 DPH and finally, 900–1000% after 25 DPH.
Water quality parameters during the larval rearing trials were maintained as follow: 25.5 ± 1C temperature;
6.0–10.5 mg/L dissolved oxygen; 36-ppt salinity. The photoperiod was maintained 16-h light: 8-h dark, with excep-
tion of four 0.4 m
3
tanks that had extended light period (McGuigan et al., 2021). Surface skimming was performed
manually with paper towels regularly throughout the day, as well as with the use of surface skimmers, which were
cleaned every 30 min after feeding to remove oil and particulate buildup. After 6 DPH all tanks were siphoned once
a day to remove detritus from the bottom.
As an effort to reduce bacterial load of the tanks, a passive transfer was done in each tank during the first days
of introduction of Artemia sp. The passive transfer consisted of connecting two tanks by a 5 cm diameter flexible
hose and siphoning water from one tank (with larvae) to the other tank. The larvae were passively transferred over a
period of 12 h to a clean and disinfected tank. According to Stuart et al. (2016) passive transfer can considerably
reduce the bacterial load in Seriola lalandi larval rearing tanks, contributing directly to improved survival. A positive
effect was also observed when using this technique in high-performance species as cobia and mahi-mahi (Benetti
et al., 2008; Stieglitz et al., 2017).
The larval rearing protocol established for both production trials is presented in Figure 1.
4BUCHALLA ET AL.
2.3 |Live feed production and feeding regime
The s-strain rotifer (B. rotundiformis) used in this study were cultured in a semi-continuous indoor system consisting
of two 1 m
3
cylindrical tanks and three 0.4 m
3
conical tanks. The initial density of the culture was 1000 rotifer/ml
and after a 3-day cycle when the rotifers were harvested, the density was 1800–2000 rotifer/ml. Rotifers were fed
with OriOne (Skretting, Tooele, UT), and 24 h prior to larval feeding, rotifers were harvested, rinsed with filtered sea-
water, placed in the 0.4 m
3
conical tanks, and fed with another enrichment diet developed at UMEH (Table 1) con-
taining Rotigreen, N-Rich (Reed Mariculture Inc., Campbell, CA), Protein Plus, AlgaMac 3050, Astaxanthin
(Aquafauna Bio-Marine Inc., Hawthorne, CA), and yeast (Lesaffre Corporation, Milwaukee, WI). This enrichment mix
(OriOne and UMEH enrichment diet) allows the rotifers to assimilate significant amounts of fatty acids in the tissue
and guts, which is essential for marine fish larvae (Tocher, 2010; Watanabe, 1993). Taurine, a component that has
been proven to impart positive effects on the growth and survival rate of marine fish larvae (Chen et al., 2004,2005;
Matsunari et al., 2013; Salze et al., 2012), was also utilized in the rotifer enrichment for the last 24 h prior to larval
FIGURE 1 Rearing protocol of red snapper Lutjanus campechanus larvae.
TABLE 1 UMEH culture and enrichment protocol for s-strain rotifers (Brachionus rotundiformis).
Culture day Culture density (rotifers/ml) Media diet Feeding rate (g mill rot
1
)
Day 1 1000 OriOne 0.45–0.55
Day 2 1200–1500 OriOne 0.35–0.45
Day 3 1500–2000 OriOne 0.3–0.25
24 h UMEH enrichment 1000 Rotigreen (20%) N-Rich (5%) Protein
Plus (2.5%) AlgaMac (30%)
Astaxantin (2.5%) Yeast (40%)
0.3
BUCHALLA ET AL.5
feeding at a rate of 4 g/L. The addition of dissolved taurine in the rotifers culture can result in internal taurine con-
centrations identical to that of wild copepods (Hawkyard et al., 2016).
Rotifers were harvested before each feeding period, at 06:00 a.m., 10:30 a.m., 01:30 p.m., 04:30 p.m., and
07:30 p.m., after counting the residual quantities of rotifers in each larval tank. The number of rotifers fed to each
tank was calculated to maintain a concentration of 15 rotifers/ml in the first day of feeding, 20 rotifers/ml between
the 4–5 DPH, and 25 rotifers/ml from 6 to 20 DPH. After 20 DPH, the number of rotifers was gradually decreased
until 25 DPH, when the fish were fed exclusively Artemia sp. metanauplii. Rotigreen algae in paste was used as green
water in the larval tanks, added prior to each rotifer feeding in concentrations of 6–20 ml of algae/m
3
, depending on
the intensity of water exchange, to maintain a concentration of 500,000–700,000 cells/ml in the rearing tanks.
Green water has been widely used in larval fish aquaculture as a form of mitigating light-intensity and enabling effi-
cient establishment of exogenous feeding, as well as a nutritional supplement for live feed (Schwarz et al., 2008; Van
der Meeren et al., 2007; Wang et al., 2019).
Artemia sp. nauplii and metanauplii were supplied to the larvae from 18 DPH until 35 DPH. Recently hatched
nauplii were provided for the first 3 days, and enriched metanauplii for the rest of the period. Artemia sp. metanauplii
were enriched with a diet developed at UMEH containing 85% Algamac 3050 (Aquafauna Bio-Marine Inc., Haw-
thorne, CA), 10% N-rich, 5% Astaxanthin, 1 g/L of taurine, and 0.5 g/L of sodium bicarbonate. A commercial water-
conditioner Sanocare Ace (INVE Aquaculture Inc., Salt Lake City, UT) was used in both the hatching and enrichment
process. Artemia sp. used for the daily feedings were maintained in a cold storage container with temperatures ranging
from 8–12C. The Artemia sp. were fed to the larvae at 07:00 a.m., 09:00 a.m., 12:00 p.m., 02:00 p.m., 04:00 p.m.,
07:00 p.m., starting with a concentration of 0.05 Artemia sp/ml per feeding at 18 DPH, reaching 7 Artemia sp/ml per
feeding at 30 DPH. At this point, co-feeding with dry feed began and the amount of Artemia sp. that were supplied
for each feeding gradually decreased until juveniles were completely weaned onto dry feeds.
An artificial diet of 350-μsized crumble with 56.3% of crude protein (Otohime; Reed Mariculture Inc., Campbell,
CA) was first introduced ad libitum at 30 DPH, and the particle size of inert diet was gradually increased until the fish
were completely weaned at 35 DPH. During the weaning, the inert diet was supplied 30 min before each feeding of
Artemia sp., and after the co-feeding concluded, it was supplied each hour.
The feeding protocol for red snapper larval rearing developed in this study is presented in Figure 2.
2.4 |Sample and data analysis
Larvae were sampled daily during the first 6 days of feeding. In the first larval rearing trial, an estimation of survival
rate was determined at 12 DPH by collecting three subsamples of tank water to estimate the total number of larvae
in the tank. A 5 cm diameter straight pipe with a valve was submerged in each tank to collect the samples from the
entire water column (vertical sample). The contents of the samples were poured over a 200-μfilter, and the total
number of larvae were counted. The result was expressed as a mean of three samples. The survival rate at this period
(pre-flexion stage) was used to determine the adequacy of rotifers as first prey for red snapper larvae. In the second
larval rearing trial, the survival rate was determined at 20 DPH by similar methodology. The estimation of the sur-
vival rate at this point could be useful for understanding the most important periods of mortality during larval rearing
that are not related to first feeding.
At 40 DPH, juveniles were collected from all tanks and transferred to different nursery systems. In the first larval
rearing trial, all larvae were manually counted during transfer, and in the second larval rearing trial, as the tanks had
higher density, fish were sedated using clove oil at a concentration of 2 ppm to reduce handling stress during trans-
fer; and batched weighed with the use of fish nets and 15 L buckets to obtain the total biomass in each tank.
Three samples of 50 juveniles per sample were collected and weighed to obtain the average weight per fish. The
total number of harvested fish from each tank was obtained by dividing the total biomass harvested by the average
juvenile weight. Measurements of standard length (mm) were made at 3 DPH and at time of transfer with an optical
microscope (Leica Microscopes CME; Buffalo, NY).
6BUCHALLA ET AL.
Productivity (juveniles/L) and final survival were calculated for each tank and trial. Final survival rates from each
trial were arcsine-transformed and a student's t-test (Microsoft Excel, 2016) was used to compare mean survival
rates among the two trials (Zar, 1996). The coefficient of variation (CV) was calculated for the mean percentage final
survival of each trial.
3|RESULTS
3.1 |Egg production
Over the course of the spawning season reported in this study, a total of 6.44 million eggs was collected from 23 via-
ble natural spawns from the broodstock cohort, with fertility rate averaging 68.0 ± 6.35%. For the larval rearing trials
reported in present study, 1.01 million fertilized eggs were used: a total of 135,000 eggs for the first larval rearing
trial and 877,000 eggs for the second larval rearing trial.
The average diameter of the eggs was 847.75 ± 5.78 μm, and the mean oil droplet diameter was 151.11 ± 1.36 μm.
For the two spawns used in the larval rearing trials, the mean fertilization rate was 95.50%, and the mean hatch
rate was 82.22%. At 25.5C, the incubation period was 24 h. The standard length of newly hatched larvae aver-
aged2.11±0.03mm.
3.2 |Larval and juvenile production
The larvae exhibited fully pigmented eyes and open mouths by the end of 2 DPH. At the start of the first feeding
(at 3 DPH), the yolk-sac reserves and oil globules were almost fully consumed. At the end of the first day of feeding,
60% of larvae exhibited enriched rotifer content in their guts. At 4 DPH, all larvae had rotifers in their guts.
FIGURE 2 Feeding protocol of red snapper Lutjanus campechanus larvae in culture conditions.
BUCHALLA ET AL.7
The formation of the swim bladder started at 6 DPH, and its inflation occurred until 12 DPH, when swim bladder
inflation was >95% in all larval rearing tanks. The survival rate of the tanks calculated at 12 DPH averaged 66.09
± 0.08% (Table 2), and at 20 DPH averaged 23.32 ± 0.02% (Table 3).
A delicate period was observed with the introduction of the Artemia sp. at 18 DPH, and persisted
until 30 DPH, when the larvae are going through metamorphosis. Settlement behavior at the bottom of the
tank became apparent, and it was possible to alleviate this behavior throughout metamorphosis by manipulat-
ing light intensity and aeration. Overall, the larvae exhibited considerable high mortality during this critical
phase.
After 30 DPH, cannibalistic behavior was very evident. Size heterogeneity in the larvae and juveniles was
observed as early as 12 DPH, but it was also noted that the cannibalism was not solely a size related issue, as a gen-
eral aggressive behavior was characteristically of larvae of red snapper at this stage. This behavior was more evident
in tanks with higher densities. In the present study, higher mortalities were related to cannibalism after the period of
metamorphosis, persisting until 40 DPH.
A total of 5253 juveniles were harvested at 40 DPH from the eight 0.4 m
3
tanks, with survival rates ranging
from 1.54% to 11.27%. The average survival rate was 4.61 ± 0.01% at the same age (Figure 3), and calculated
CV was 0.64. The mean final density of this larval rearing trial was 1.64 ± 0.40 juveniles/L. A total of 26,596
juveniles were produced in the four 2.4 m
3
tanks with survival rates at 40 ± 2.8 DPH ranging from 3.10% to
5.56%. The average survival rate was 3.95 ± 0.01%, and calculated CV was 0.28. Fish averaged 0.35 ± 0.02 g in
body weight and 28.83 ± 0.06 mm in standard length, and the mean final density was 3.69 ± 0.44 juveniles/L.
Mean final survival rates were not statistically different among the two larval rearing trials (student's t-test,
p> 0.05). In total, 31,849 early-juveniles were harvested at 40 ± 2.8 DPH across all tanks, with a mean final sur-
vival of 4.43% ± 0.01, and mean final density of 2.20 ± 0.42 juveniles/L.
4|DISCUSSION
Enriched s-strain rotifers were successfully used as the sole first-feeding prey item for L. campechanus larvae, leading
to pilot-scale production quantities of fully weaned juveniles that meet or exceed those reported in other studies
using copepod nauplii for first feeding (Bootes, 1998; Ogle & Lotz, 2006). In addition, when tracking larval growth
and development from first-feeding larvae to early-juveniles, the results achieved here mirrored the study conducted
TABLE 2 Summary of stocking density, survival rates, number of larvae and juveniles produced of red snapper
Lutjanus campechanus during Trial 1.
Tank Volume (m
3
)
Larvae
stocked 1 DPH
Number of
larvae at 12 DPH
Survival at
12 DPH
Number of
juveniles
at 40 DPH
Survival
at 40 DPH Juveniles/L
S1
a
0.4 14,250 2420 16.98% 689 4.84% 1.72
S2
a
0.4 14,250 9696 68.04% 950 6.67% 2.37
S3
a
0.4 14,250 10,181 71.45% 418 2.93% 1.04
S5
a
0.4 14,250 10,666 74.85% 337 2.36% 0.84
S6 0.4 14,250 12,120 85.05% 629 4.41% 1.57
S7 0.4 14,250 13,090 91.86% 1606 11.27% 4.01
S8 0.4 14,250 9212 64.65% 220 1.54% 0.55
S10 0.4 14,250 7957 55.84% 404 2.84% 1.01
Mean ± SEM 66.09 ± 0.08 4.61% ± 0.01 1.64 ± 0.40
a
Tanks had photoperiod of 24 h light until 12 DPH.
8BUCHALLA ET AL.
TABLE 3 Summary of stocking density, survival rates, number of larvae and juveniles produced, and size parameters of red snapper Lutjanus campechanus during Trial 2.
Tank Volume (m
3
)
Larvae stocked
1 DPH
Number of larvae
at 20 DPH
Survival
at 20 DPH
Number of
juveniles at 40 DPH
Survival
at 40 DPH
Mean weight
at 40 DPH (g)
Standard length
at 40 DPH (mm) Juveniles
A1 2.4 108,000 30,000 27.78% 9211
a
3.10% 0.32 28 3.83
A2 2.4 188,800 40,000 21.19%
A3 2.4 188,800 40,000 21.19% 10,503 5.56% 0.4 30 4.37
A4 2.4 216,000 50,000 23.15% 6882 3.19% 0.34 28.5 2.86
Mean ± SEM 23.32 ± 0.02 3.95 ± 0.01 0.35 ± 0.02 28.83 ± 0.60 3.69 ± 0.44
a
Tank A2 was combined with A1 before the final count at 40 DPH.
BUCHALLA ET AL.9
by Drass et al. (2000) which fed copepods as first prey item, suggesting that the larvae in the current study did not
suffer from any nutritional deficiencies as a result of diet. Such results suggest that providing high-quality enriched
s-strain rotifers can result in successful first feeding of this species (60% feeding success during the first day of feed-
ing, and 100% during the second day of feeding) and establish a sufficient diet for downstream success throughout
the entire larval period.
In many studies involving L. campechanus, major bottlenecks were reported during the first days of larval rearing
related with the first feeding, and high mortalities were observed as a consequence after 8 DPH (Bardon-
Albaret, 2014; Bardon-Albaret et al., 2013; Bardon-Albaret & Saillant, 2017; Ogle & Lotz, 2006; Saillant et al., 2013;
Williams et al., 2004). Such issues were not observed in the present study. The estimated survival rate (Figure 3)at
12 DPH (66.09 ± 0.08%) was high when compared to survival rates at this stage of development in other challenging
marine fish species (Alvarez-Lajonchère et al., 2012; Gutiérrez-Sigeros et al., 2018; Ogle & Lotz, 2006) demonstrating
the success in the first feeding using exclusively rotifers. When compared with the mean final survival rate (4.43
± 0.01%) at 40 DPH, results indicate that the high mortalities during larval rearing were observed in three further
phases: (a) metamorphosis, after 20 DPH, (b) introduction of Artemia sp. and (c) weaning and cannibalism after
30 DPH. These critical periods were also reported in other snapper species (Gutiérrez-Sigeros et al., 2018; Leu
et al., 2003; Watanabe et al., 1998).
These limitations can be avoided and further work with red snapper larval rearing should consider: (a) alleviating
the stress of the larvae through manipulation of light, water current, aeration, and improved larval nutrition
(Ibarra-Castro et al., 2020); (b) providing a high-quality, clean and disinfected Artemia sp., since it can be the source
of introduction of bacteria in the culture environment (Høj et al., 2009); and (c) improving feeding and size-grading
protocol after weaning to avoid the cannibalism associated with size heterogeneity of the cohort (Folkvord, 1991).
Mean final survival rates were not statistically different among the two larval rearing trials, suggesting that final
survival was not density-dependent, and initial stock densities ranging 35–75 larvae/L can be suitable for red snap-
per larval rearing (Figure 3).
FIGURE 3 Survival rates of Lutjanus campechanus for Trial 1 (initial stock density of 35 larvae L
1
), and for Trial
2 (initial stock density of 75 larvae L
1
). Bars indicate mean percentage survival rate from each trial at different
stages of development, and circles indicate survival of individual tanks. Error bars are 2 SEM.
10 BUCHALLA ET AL.
Although copepods have been successfully used for a variety of snapper species that also exhibit small mouth
gapes, their culture is reportedly more complex, expensive, and require more production area than rotifer culture
(Saillant et al., 2013; Stottrup, 2000), especially when considering the production of commercial amounts of live prey.
Consequentially, the exclusive use of rotifers as first-feeding prey items for this species is a very encouraging result
for the development of commercial-scale red snapper aquaculture industry.
While no direct comparison was made between the use of rotifers and copepods during the first feeding of red
snapper in this study, future research should consider investigating differences that may occur with the use of each
prey type during the larval rearing of this species. Furthermore, integrating the use of rotifers and copepods during
the first feeding could be an alternative for small-mouth marine fish species, and likely optimize results. However, as
private production companies raising red snapper seek immediate methods to improve larval production, the proto-
col presented in this study represents an important alternative that does not necessitate copepod culture.
The development of a reliable and intensive protocol for larval rearing production of a species is one of the first
steps in realizing commercial-scale culture feasibility. This study builds upon results of previous trials conducted on
this species. As reported, the exclusive use of rotifers as first-feeding prey for red snappers overcomes a significant
bottleneck for the reliable mass production of this species and makes an important contribution towards a collective
effort to support the development of commercial-scale aquaculture of this iconic species.
ACKNOWLEDGMENTS
This research was supported by a grant from the National Oceanic and Atmospheric Administration and Florida Sea
Grant [NOAA-OAR-SG-2017-2005176]. The authors acknowledge the contribution and dedication of the University
of Miami Experimental Hatchery and Aquaculture Program students, staff, and volunteers. Handling and animal use
and welfare of in the present study were in compliance with the Institutional Animal Care and Use Committee of the
University of Miami protocol #18-052LF.
DATA AVAILABILITY STATEMENT
The data that support the findings of this study are available from the corresponding author upon reasonable
request.
ORCID
Yole Buchalla https://orcid.org/0000-0003-0600-6131
John D. Stieglitz https://orcid.org/0000-0003-3926-4364
REFERENCES
Alvarez-Lajonchère, L., Abdo De La Parra, M. I., Rodríguez Ibarra, L. E., Velasco Blanco, G., Puello-Cruz, A. C., González
Rodríguez, B., & Ibarra Castro, L. (2012). The scale-up of spotted rose snapper, Lutjanus guttatus, larval rearing at
Mazatlán, Mexico. Journal of the World Aquaculture Society,43(3), 411–422. https://doi.org/10.1111/j.1749-7345.2012.
00573.x
Alvarez-Lajonchère, L., Sánchez, L. P., Molej
on, O. H., & G
omez, E. T. (1996). Mass production of striped patao Eugerres
brasilianus juveniles in Cuba. Journal of the World Aquaculture Society,27(3), 347–352. https://doi.org/10.1111/j.1749-
7345.1996.tb00618.x
Arnold, C. R., Wakeman, J. M., Williams, T. D., & Treece, G. D. (1978). Spawning of red snapper (Lutjanus campechanus)in
captivity. Aquaculture,15(3), 301–302. https://doi.org/10.1016/0044-8486(78)90040-6
Bardon-Albaret, A. B. (2014). Characterization of the early development and quality of red snapper (Lutjanus campechanus) eggs
and larvae in aquaculture conditions. Dissertations. University of Southern Mississippi. 749. The University of Southern
Mississippi. Retrieved from https://aquila.usm.edu/dissertations/749
Bardon-Albaret, A., Brown-Peterson, N. J., Lemus, J. T., Apeitos, A., & Saillant, E. A. (2013). A histological study of gameto-
genesis in captive red snapper Lutjanus campechanus.Aquaculture Research,1–8, 901–908. https://doi.org/10.1111/are.
12244
Bardon-Albaret, A., & Saillant, E. (2017). Egg quality traits and predictors of embryo and fry viability in red snapper Lutjanus
campechanus.Aquaculture Reports,7,48–56. https://doi.org/10.1016/j.aqrep.2017.05.004
BUCHALLA ET AL.11
Benetti, D., Orhun, M., Sardenberg, B., O'Hanlon, B., Welch, A., Hoenig, R., Zink, I., Rivera, J., Denlinger, B., Bacoat, D.,
Palmer, K., & Cavalin, F. (2008). Advances in hatchery and grow-out technology of cobia Rachycentron canadum
(Linnaeus). Aquaculture Research,39(7), 701–711. https://doi.org/10.1111/j.1365-2109.2008.01922.x
Blanco, E., Reglero, P., Ortega, A., Folkvord, A., de la Gándara, F., Hernández de Rojas, A., & Moyano, M. (2020). First esti-
mates of metabolic rate in Atlantic bluefin tuna larvae. Journal of Fish Biology,97(5), 1296–1305. https://doi.org/10.
1111/jfb.14473
Bootes, K. L. (1998). Culture and description of larval red snapper, Lutjanus campechanus. Master's thesis. Alburn University.
Castaños, M. T. (1997). A hatchery rearing method for the mangrove red snapper. SEAFDEC Asian Aquaculture,19(1), 1–5.
Chávez, E. A., Santamaria Miranda, A., & Ponce Palafox, J. T. (2014). Bioeconomic analysis of three west American snappers
(Lutjanus spp) with aquaculture potential. ARAMARA Repositorio Institucional, Universidad Aut
onoma de Nayarit.
Chen, J. N., Takeuchi, T., Takahashi, T., Tomoda, T., Koiso, M., & Kuwada, H. (2004). Effect of rotifers enriched with taurine
on growth and survival activity of red sea bream Pagrus major larvae. Bulletin of the Japanese Society of Scientific Fisheries,
70(4), 542–547.
Chen, J. N., Takeuchi, T., Takahashi, T., Tomoda, T., Koiso, M., & Kuwada, H. (2005). Effect of rotifers enriched with taurine
on growth in larvae of Japanese flounder Paralichthys olivaceus.Bulletin of the Japanese Society of Scientific Fisheries,
71(3), 342–347.
DiMaggio, M. (2020). Overview of commonly cultured marine ornamental fish. EDIS,2020(3), 7. https://doi.org/10.32473/
edis-fa226-2020
Doi, M., Ohno, A., Taki, Y., Singhagraiwan, T., & Kohno, H. (1997). Nauplii of the calanoid copepod, Acartia sinjiensis as an ini-
tial food organism for larval red snapper, Lutjanus argentimaculatus.Aquaculture Science,45(1), 31–40. https://doi.org/
10.11233/aquaculturesci1953.45.31
Drass, D. M., Bootes, K. L., Lyczkowski-Shultz, J., Comyns, B. H., Holt, J. G., Riley, C. M., & Phelps, R. P. (2000). Larval devel-
opment of red snapper, Lutjanus campechanus, and comparisons with co-occurring snapper species. Fishery Bulletin,98,
507–527.
Duray, M. N., Alpasan, L. G., & Estudillo, C. B. (1996). Improved hatchery rearing of mangrove red snapper, Lutjanus
argentimaculatus, in large tanks with small rotifer (Brachionus plicatilis) and Artemia. Israeli Journal of Aquaculture,48(3),
123–132.
Duray, M. N., & Kohno, H. (1990). Development of mouth width and larval growth in three marine fish species. Philippine
Journal of Science,119(3), 237–245.
Estrada-Godínez, J. A., Moreno-Figueroa, L. D., Maldonado-García, M., Pérez-Urbiola, J. C., Romero-Rodríguez, J., & Audelo-
Naranjo, J. M. (2015). Influence of the temperature on the early larval development of the Pacific red snapper, Lutjanus
peru (Nichols & Murphy, 1922). Latin American Journal of Aquatic Research,43(1), 137–145. https://doi.org/10.3856/
vol43-issue1-fulltext-12
Folkvord, A. (1991). Growth, survival and cannibalism of cod juveniles (Gadus morhua): Effects of feed type, starvation and
fish size. Aquaculture,97(1), 41–59. https://doi.org/10.1016/0044-8486(91)90278-F
Geng, J., Belfranin, C., Zander, I. A., Goldstein, E., Mathur, S., Lederer, B. I., Benvenuti, R., & Benetti, D. D. (2019). Effect of
stocking density and feeding regime on larval growth, survival, and larval development of Japanese flounder, Paralichthys
olivaceus, using live feeds. Journal of the World Aquaculture Society,50(2), 336–345. https://doi.org/10.1111/jwas.
12563
Gutiérrez-Sigeros, I., Ibarra-Castro, L., Alvarez-Lajonchère, L., & Sanchez-Zamora, A. (2018). Natural spawning and scaling-up
of yellowtail snapper (Ocyurus chrysurus): Larval rearing for the mass production of juveniles. Aquaculture,491, 252–
257. https://doi.org/10.1016/j.aquaculture.2018.03.048
Hastey, R., Phelps, R., Davis, A., & Cummins, K. (2013). Augmentation of free amino acids in eggs of red snapper, Lutjanus
campechanus as part of the induced spawning protocol. Aquaculture Research,1–8, 283–290. https://doi.org/10.1111/
are.12184
Hawkyard, M., Stuart, K., Langdon, C., & Drawbridge, M. (2016). The enrichment of rotifers (Brachionus plicatilis) and Artemia
franciscana with taurine liposomes and their subsequent effects on the larval development of California yellowtail
(Seriola lalandi). Aquaculture Nutrition,22(4), 911–922. https://doi.org/10.1111/anu.12317
Høj, L., Bourne, D. G., & Hall, M. R. (2009). Localization, abundance and community structure of bacteria associated with
artemia: Effects of nauplii enrichment and antimicrobial treatment. Aquaculture,293(3–4), 278–285. https://doi.org/10.
1016/j.aquaculture.2009.04.024
Ibarra-Castro, L., Ochoa-Boj
orquez, M. O., Sánchez-Téllez, J. L., Rojo-Cebreros, A. H., & Alvarez-Lajonchère, L. (2020). A
new efficient method for the mass production of juvenile spotted rose snapper Lutjanus guttatus.Aquaculture Reports,
18, 100550. https://doi.org/10.1016/j.aqrep.2020.100550
Kaji, T., Tanaka, M., Oka, M., Takeuchi, H., Ohsumi, S., Teruya, K., & Hirokawa, J. (1999). Growth and morphological develop-
ment of laboratory-reared yellowfin tuna Thunnus albacares larvae and early juveniles, with special emphasis on the
digestive system. Fisheries Science,65(5), 700–707. https://doi.org/10.2331/fishsci.65.700
12 BUCHALLA ET AL.
Lee, I. S., Ohs, C. L., Broach, J. S., DiMaggio, M. A., & Watson, C. A. (2018). Determining live prey preferences of larval orna-
mental marine fish utilizing fluorescent microspheres. Aquaculture,490, 125–135. https://doi.org/10.1016/j.
aquaculture.2018.01.035
Leu, M.-Y., Chen, I., & Fang, L. S. (2003). Natural spawning and rearing of mangrove red snapper, Lutjanus argentimaculatus,
larvae in captivity. The Israeli Journal of Aquaculture,55,22–30.
Margulies, D., Wexler, J. B., Bentler, K. T., Suter, J. M., Masuma, S., Tezuka, N., Teruya, K., Oka, M., Kanematsu, M., &
Nikaido, H. (2001). Food selection of yellowfin tuna, Thunnus albacares, larvae reared in the laboratory. Inter-American
Tropical Tuna Commission Bulletin,22,9–51.
Matsunari, H., Hashimoto, H., Iwasaki, T., Oda, K., Masuda, Y., Imaizumi, H., Teruya, K., Furuita, H., Yamamoto, T.,
Hamada, K., & Mushiake, K. (2013). Effect of feeding rotifers enriched with taurine on the growth and survival of larval
amberjack Seriola dumerili.Fisheries Science,79(5), 815–821. https://doi.org/10.1007/s12562-013-0657-y
McGuigan, C. J., Buchalla, Y., Darville, K. G., Tudela, C. E., Stieglitz, J. D., Hoenig, R. H., & Benetti, D. D. (2021). Effect of
replacing darkness with dim light in the larviculture of red snapper, Lutjanus campechanus.Aquaculture Reports,20, 100762.
https://doi.org/10.1016/j.aqrep.2021.100762
NOAA. (2020). Red snapper. Retrieved from https://www.fisheries.noaa.gov/species/red-snapper
Ogle, J. T., Lemus, J. T., Nicholson, L. C., Barnes, D. N., & Lotz, J. M. (2005). Characterization of an extensive zooplankton
culture system coupled with intensive larval rearing of red snapper Lutjanus campechanus.InCopepods in aquaculture
(pp. 225–244). Blackwell Publishing Ltd. IA. https://doi.org/10.1002/9780470277522.ch17
Ogle, J. T., & Lotz, J. M. (2006). Characterization of an experimental indoor larval production system for red snapper. North
American Journal of Aquaculture,68(1), 86–91. https://doi.org/10.1577/A05-029.1
Papanikos, N., Phelps, R. P., Davis, D. A., Ferry, A., & Maus, D. (2008). Spontaneous spawning of captive red snapper,
Lutjanus campechanus, and dietary lipid effect on reproductive performance. Journal of the World Aquaculture Society,
39(3), 324–338. https://doi.org/10.1111/j.1749-7345.2008.00171.x
Phelps, R. P., Sumiarsa, G. S., Lipman, E. E., Lan, H. P., Moss, K. K., & Davis, A. D. (2005). Intensive and extensive production
techniques to provide copepod nauplii for feeding larval red snapper Lutjanus campechanus.InCopepods in aquaculture
(pp. 151–168). Blackwell Publishing Ltd. IA.
Rabalais, N. N., Rabalais, S. C., & Arnold, C. R. (1980). Description of eggs and larvae of laboratory reared red snapper,
Lutjanus campechanus.American Society of Ichthyologists and Herpethologists,4, 704–708 Retrieved from http://www.
jstor.org/stable/1444447
Rhodes, M. A., & Phelps, R. P. (2008). Evaluation of the ciliated protozoa, Fabrea salina as a first food for larval red snapper,
Lutjanus campechanus in a large-scale rearing experiment. Journal of Applied Aquaculture,20(2), 120–133. https://doi.
org/10.1080/10454430802197383
Saillant, E. A., Leclercq, E., Bardon-Albaret, A., Sarkisian, B., Apeitos, A., Brown-Peterson, N. J., Lemus, J. T., Buentello, A.,
Pohlenz, C., & Gatlin, D. M. (2013). Development of aquaculture of the red snapper Lutjanus campechanus: Research on lar-
val nutrition (pp. 352–356). 65th Gulf and Caribbean Fisheries Institute.
Salze, G., McLean, E., & Craig, S. R. (2012). Dietary taurine enhances growth and digestive enzyme activities in larval cobia.
Aquaculture,362,44–49.
Sawada, Y., Okada, T., Miyashita, S., Murata, O., & Kumai, H. (2005). Completion of the Pacific bluefin tuna Thunnus orientalis
(Temminck et Schlegel) life cycle. Aquaculture Research,36(5), 413–421. https://doi.org/10.1111/j.1365-2109.2005.
01222.x
Schipp, G. (2006). The use of calanoid copepods in semi-intensive, tropical marine fish larviculture. Avances en Nutrici
on
Acuícola,160,84–94.
Schwarz, M. H., Craig, S. R., Delbos, B. C., & McLean, E. (2008). Efficacy of concentrated algal paste during greenwater phase
of cobia larviculture. Journal of Applied Aquaculture,20(4), 285–294. https://doi.org/10.1080/10454430802498583
Stein, M. S., Margulies, D., Wexler, J. B., Scholey, V. P., Ryo, K., Honryo, T., Sasaki, T., Guillen, A., Agawa, Y., & Sawada, Y.
(2018). A comparison of the effects of two prey enrichment media on growth and survival of Pacific bluefin tuna,
Thunnus orientalis, larvae. Journal of the World Aquaculture Society,49(1), 240–255. https://doi.org/10.1111/jwas.12478
Stieglitz, J. D., Benetti, D. D., Hoenig, R. H., Sardenberg, B., Welch, A. W., & Miralao, S. (2012). Environmentally conditioned,
year-round volitional spawning of cobia (Rachycentron canadum) in broodstock maturation systems. Aquaculture
Research,43(10), 1557–1566. https://doi.org/10.1111/j.1365-2109.2011.02961.x
Stieglitz, J. D., Hoenig, R. H., Kloeblen, S., Tudela, C. E., Grosell, M., & Benetti, D. D. (2017). Capture, transport, prophylaxis,
acclimation, and continuous spawning of Mahi-mahi (Coryphaena hippurus) in captivity. Aquaculture,479,1–6. https://
doi.org/10.1016/j.aquaculture.2017.05.006
Stottrup, J. G. (2000). The elusive copepods: Their production and suitability in marine aquaculture. Aquaculture Research,
31(8–9), 703–711.
Stuart, K., Rotman, F., & Drawbridge, M. (2016). Methods of microbial control in marine fish larval rearing: Clay-based tur-
bidity and passive larval transfer. Aquaculture Research,47(8), 2470–2480. https://doi.org/10.1111/are.12696
BUCHALLA ET AL.13
Tocher, D. R. (2010). Fatty acid requirements in ontogeny of marine and freshwater fish. Aquaculture Research,41(5), 717–
732. https://doi.org/10.1111/j.1365-2109.2008.02150.x
Van der Meeren, T., Mangor-Jensen, A., & Pickova, J. (2007). The effect of green water and light intensity on survival,
growth, and lipid composition in Atlantic cod (Gadus morhua) during intensive larval rearing. Aquaculture,265(1–4),
206–217.
Wang, J., Shu, X., & Wang, W. X. (2019). Micro-elemental retention in rotifers and their trophic transfer to marine fish lar-
vae: Influences of green algae enrichment. Aquaculture,499, 374–380. https://doi.org/10.1016/j.aquaculture.2018.
09.066
Watanabe, T. (1993). Importance of docosahexaenoic acid in marine larval fish. Journal of the World Aquaculture Society,
24(2), 152–161. https://doi.org/10.1111/j.1749-7345.1993.tb00004.x
Watanabe, W. O., Benetti, D. D., Feeley, M. W., Davis, A. D., & Phelps, R. P. (2005). Status of artificial propagation of mut-
ton, yellowtail, and red snapper (family Lutjanidae). In American Fisheries Society Symposium (vol. 46) (pp. 517–540).
American Fisheries Society.
Watanabe, W. O., Ellis, E. P., Ellis, S. C., Chaves, J., Manfredi, C., Hagood, R. W., Sparsis, M., & Arneson, S. (1998). Artificial
propagation of mutton snapper Lutjanus analis, a new candidate marine fish species for aquaculture. Journal of the World
Aquaculture Society,29(2), 176–187. https://doi.org/10.1111/j.1749-7345.1998.tb00977.x
Williams, K., Papanikos, N., Phelps, R. P., & Shardo, J. D. (2004). Development, growth, and yolk utilization of hatchery-
reared red snapper Lutjanus campechanus larvae. Marine Ecology Progress Series,275, 231–239. https://doi.org/10.
3354/meps275231
Wullur, S., Sakakura, Y., & Hagiwara, A. (2011). Application of the minute monogonont rotifer Proales similis de Beauchamp
in larval rearing of seven-band grouper Epinephelus septemfasciatus.Aquaculture,315(3–4), 355–360.
Zar, J. H. (1996). Biostatistical analysis. Printice-Hall, Inc.
Zavala-Leal, I., Dumas, S., L
opez-Villegas, E. O., Peña, R., Contreras-Olguín, M., Flores-Montijo, L., & De La Cruz-Agüero, J.
(2015). Structural development of Pacific red snapper Lutjanus peru from hatching to the onset of first feeding. Aquacul-
ture Research,46(5), 1162–1174. https://doi.org/10.1111/are.12272
How to cite this article: Buchalla, Y., McGuigan, C. J., Stieglitz, J. D., Hoenig, R. H., Tudela, C. E., Darville,
K. G., Ibarra-Castro, L., & Benetti, D. (2023). Advancements in hatchery production of red snapper Lutjanus
campechanus: Exclusive use of small strain rotifers as initial prey for larval rearing. Journal of the World
Aquaculture Society,1–14. https://doi.org/10.1111/jwas.12992
14 BUCHALLA ET AL.