




































 Agricultural Science; Vol. 4, No. 2; 2022 
ISSN 2690-5396   E-ISSN 2690-4799 

https://doi.org/10.30560/as.v4n2p79 

79                             Published by IDEAS SPREAD 
 

Effect of Color During Transport and Anesthetic Efficacy of Alcoholic 
Drink, 2-Phenoxyethanol, Clove Oil, MS-222, and Benzocaine in 

Silver Therapon, Leiopotherapon plumbeus (Kner 1864) 
Mark June S. Consigna1 & Mark Nell C. Corpuz1 

1 Center for Research on Aquaculture and Aquatic Resources in Brackishwater Systems, Institute of Fisheries and 
Aquatic Sciences, Bataan Peninsula State University, Orani, Bataan, Philippines 
Correspondence: Mark Nell C. Corpuz, Center for Research on Aquaculture and Aquatic Resources in 
Brackishwater Systems, Institute of Fisheries and Aquatic Sciences, Bataan Peninsula State University, Orani, 
Bataan, Philippines. E-mail: mnccorpuz@bpsu.edu.ph 
 
Received: November 17, 2022   Accepted: December 9, 2022   Online Published: December 31, 2022 
 
The research is financed by Bataan Peninsula State University, Research and Development Office. 
 
Abstract 
The study evaluated the survivability of silver therapon, Leiopotherapon plumbeus (Kner, 1864) in different color 
containers during transport and the efficacy of five anesthetic agents [alcoholic drink, 2-phenoxyethanol, clove oil, 
tricaine methanesulfonate (MS-222), and benzocaine] in the induction and recovery time of L. plumbeus. Different 
colored polyethylene bags (black, red, yellow, blue, and transparent) did not influence the survival rate of fish until 
the termination of the experiment (12-h transport time). The immersion experiment used three different 
concentrations in each anesthetic agent with three replicates (ten fish specimens per replicate). Different dosages 
significantly influenced the induction time, with decreased induction efficacies in high dosages. Moreover, the 200 
ml L-1 and 300 ml L-1 alcoholic drinks anesthetized the fish specimens comparable to the induction efficacy of 
several dosages of 2-phenoxyethanol, MS-222, and benzocaine. Recovery time significantly varied among 
treatments, with a prolonged recovery period with increasing anesthetic concentrations. Regression analysis 
revealed a positive correlation between fish standard length and induction time (P < 0.05), albeit more pronounced 
in smaller dosages. Induction and recovery times were not correlated to fish size when exposed to higher dosages. 
The present finding demonstrated the anesthetic efficacy of four commercial anesthetic solutions, so as with 
alcoholic drinks with concentrations between or equal to 200 ml L-1 and 300 ml L-1. Experimental trials for fish 
euthanization and field trials are open for further investigation. 
Keywords: Ayungin, Bataan, induction time, anesthesia, recovery time 
1. Introduction 
Silver therapon (Leiopotherapon plumbeus Kner 1864), locally known as ayungin is an endemic commercial fish 
thriving in freshwaters (Paller et al. 2011) and brackishwaters of Luzon, Philippines (De Leon et al. 2017; Santos 
et al. 2020). It is regarded as an important fishery resource for subsistence fisheries which is priced from $ 4 to $ 8 
per kilogram (Corpuz and Espaldon 2021). Wild populations of freshwater fishes, however, are imperiled by 
overfishing, habitat alterations (Corpuz et al. 2015), and the presence of alien invasive species causing their natural 
populations to dwindle (Corpuz et al. 2018). To save this terapontid, conservation management initiatives and 
repopulation through habitat restoration and hormonal-induced breeding have been implemented by the Philippine 
government and other state-funded universities (Aya et al. 2015; Consigna et al. 2019).  
One of the vital factors to take into consideration for successful induced breeding is handling stress (Hseu et al. 
1998; Weber et al. 2009). It was reported that physical stress may cause breeding inefficiency and at worst, 
mortality to the fish broodstock (Coyle et al. 2004). One method to minimize stress is to anesthetize the animals, 
which results in loss of sensitivity or insensitivity and induced sleep and muscle relaxation (general anesthesia). 
Moreover, the appropriate color background is used in aquaculture particularly in the hatchery to provide the best 
fish growth performance (Imanpoor and Abdollahi 2011; Brian 2015). In the same manner as transportation, a 
specific color background of transport bags may help the fish to reduce stress and eventual mass mortality 
(Manliclic et al. 2018).  



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In aquaculture, various anesthetic agents are being used to reduce the stress on the fish and injuries during 
transportation, breeding, and fish capture (Coyle et al. 2004). There are several types of anesthesia used to lessen 
the stress of fish in performing induced breeding including tricaine methanesulphonate (hereafter denoted as MS-
222). The MS-222 is a water-soluble powdered substance that is typically buffered with sodium bicarbonate to 
lessen its acidity and is commonly delivered in a water bath (Popovic et al. 2012). Benzocaine (ethyl 
paraaminobenzoate) is known for its rapid induction and recovery times and good safety margin for several groups 
of fish (Ross and Ross 2008). Clove oil is derived from Eugenia caryophyllate tree, which contains methyleugenol, 
eugenol, and isoeugenol (Soto and Burhanuddin 1995), whereas clove oil has been widely used due to their efficacy 
and being inexpensive (Uçar and Atamanalap 2010). The 2–phenoxyethanol is a colorless oily liquid with a faint 
aromatic odor but moderately soluble in water. It is widely used for transporting live fish because it is cheap, 
reliable, and efficient and its active ingredients are ethylyn glycol monophenyl ether (Weber et al. 2009; Ucar and 
Atamanalap 2010). The alcoholic drink contains the recreational drug, ethanol which is produced through the 
fermentation of grains, fruits, or other sources of sugar. The practicality of using alcoholic drinks as anesthetic 
agents lies in the efficacy in reducing the mobility and reaction of fish to pressure, and the availability and ease of 
access in local markets. Attempts of using this substance for fish anesthesia previously showed a positive sedative 
effect in zebrafish (Hullinger 2014). However, the appropriate amounts or dosages to realize its anesthetic effect 
are yet determined for L. plumbeus. 
The present study evaluated the anesthetic efficacy of alcohol drinks, 2-phenoxyethanol, clove oil, MS-222, and 
benzocaine as anesthetic agents in L. plumbeus. Specifically, this study determined the appropriate dosages for 
anesthesia, compared the induction and recovery times across different concentrations and anesthetic solutions, 
and analyzed the correlation of induction and recovery times with fish size. Moreover, we evaluated the survival 
of L. plumbeus stocked in various colored transport containers for 12-h transport time.  
2. Method 
2.1 Fish Specimens and Acclimation 
The L. plumbeus (n = 312) were obtained from the fishponds in Orani, Bataan (14˚48.50’’ N, 120˚32.60’’ E) using 
cast nets and net traps set at the outlet of each pond. The specimens were transported at the hatchery facility of 
Bataan Peninsula State University and acclimated in three 1,200 L fiberglass tanks with a continuous supply of 
running freshwater and artificial aeration. Fish specimens were maintained for two weeks under ambient 
conditions (1:1 light-dark). During acclimation, daily monitoring of dissolved oxygen (5.30 ± 0.05 mg L-1), 
temperature (26.63 ± 0.24 °C), and pH (7.81 ± 0.01) was done. Total ammonia-nitrogen concentration was recorded 
every other day and was constantly below 0.01 mg L-1. Fish were fed ad libitum twice per day with commercial 
extruded feed (crude protein = 32%). Feeding was terminated 24 h before the experiment. No mortality was 
observed during the acclimation period. Fish specimens were size-sorted five days before the initiation of the 
experiment. Two hundred (200) unsexed fish individuals were used in the study. 
2.2 Experimental Trials 
Five anesthetics were used in this study — alcoholic drink (40% alcohol by volume, 80-proof, Ginebra San Miguel 
Inc., Philippines), 2-phenoxyethanol (ethylene glycol monophenyl ether, Sigma Aldrich Co., USA), MS-222 
(Sigma Aldrich Co, USA), clove oil (90–95% eugenol, Sigma Aldrich Co., USA) and benzocaine (ethyl 4-
aminobenzoate 99%, Sigma Aldrich Co, USA).  
The different dosages of each anesthetic agent were prepared 30 min before the actual induction experiment. The 
experiment used 3-L plastic containers filled with 1 L of water with a specific dosage of anesthetic solution. 
Artificial aeration was provided throughout the experiment. The alcoholic drink, 2-phenoxyethanol, and MS-222 
(see Table 1 for each concentration) were diluted directly into the anesthetic bath, whereas benzocaine and clove 
oil were initially dissolved in ethanol (92.8%) in a ratio of 1:9 (anesthetic to ethanol) since the two anesthetics are 
slightly soluble in water. The aliquot of the stock solutions was then used to attain the specific dosages in each 
treatment. Treatments and control groups (no anesthetic) were buffered with sodium bicarbonate to attain a pH 
level of 7.5.  
The fish individual (ten fish per treatment of each anesthetic agent) was randomly placed in an experimental 
container with a specific concentration of the anesthetic solution. Treatments for all the anesthetic agents were 
investigated. The concentrations of alcoholic drinks were 100, 200, and 300 ml L-1, while the four commercial 
anesthetic agents had 0.25, 0.50, and 1.00 ml L-1 (Coyle et al. 2004). The induction time was recorded for each the 
fish when fish displayed loss of balance, cessation of swimming, decrease in opercular rate, and no reaction to 
external stimuli (Pawar et al. 2011). Anesthetized individuals were weighed (g) and the standard length (SL) was 
measured. The SL (cm) of fish specimens was statistically homogeneous (9.07 ± 0.65 cm), ranging from 8.61 to 



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9.39 mm (Table 1), whereas weight was 12.27 ± 2.62 g, varying from 7.38 to 22.76 g. Consequently, the fish was 
placed into freshwater of similar temperatures to the experimental container. Water in the recovery container was 
constantly renewed. Recovery time was noted when the fish exhibited normal swimming and reaction to stimuli 
(Silva et al. 2012). The different stages of induction and recovery times followed the criteria proposed by Gullian 
and Villanueva (2009). Recovered specimens were brought back to the hatchery; whilst dead specimens (no vital 
signs after 30 min) were preserved in a 10% formaldehyde solution for fish collection. 
 
Table 1. Mean ± standard error in standard length (mm) of experimental Leiopotherapon plumbeus (n = 200) 

Anesthetic 
Solutions 

Standard Length (mm) 
F value p 

Control T1 T2 T3 

Alcoholic drink 9.16 ± 0.21 8.61 ± 0.24 9.04 ± 0.19 9.25 ± 0.25 2.00 0.154 
2-Phenoxyethanol 9.05 ± 0.15 8.89 ± 0.30 9.11 ± 0.13 9.12 ± 0.16 0.38 0.687 
Clove oil 8.85 ± 0.26 8.82 ± 0.28 9.39 ± 0.22 9.24 ± 0.19 1.56 0.229 
MS-222 9.24 ± 0.22 9.35 ± 0.27 9.16 ± 0.14 9.01 ± 0.21 0.61 0.548 
Benzocaine 8.98 ± 0.18 9.02 ± 0.20 9.04 ± 0.12 9.07 ± 0.29 0.01 0.986 

Treatments for alcoholic drink: T1 = 100 ml L-1, T2 = 200 ml L-1, T3 = 300 ml L-1 
Treatments for commercial anesthetic agents: T1 = 0.25 ml L-1, T2 = 0.5 ml L-1, T3 = 1.0 ml L-1 
 
2.3 Influence of Color Containers 
In separate experiment, juvenile unsexed L. plumbeus from other conditioning tanks were randomly placed in 
various colored polyethylene bags (black, red, yellow, blue, and transparent) as colored containers (25.4 x 50.8 x 
0.008 cm). The procedure was adopted from the study of Manliclic et al. (2018). Each bag contained 5 L of 
freshwater with a stocking density of one fish per liter (five fish x three replicates x five color containers = 75 fish 
individuals). The water used was obtained from the conditioning tanks. Bags were sealed after the addition of 
medical oxygen. The amount of oxygen was standardized for all treatments by ensuring that all plastic bags were 
air-filled of the same height. An improvised carrier with wheels was utilized to facilitate the simulated transport. 
The carrier was manually agitated by periodic push-and-pull actions and was operated under ambient conditions 
for 12 hours. Fish mortalities were monitored at 0-h, 6-h, and 12-h transport times.  
2.4 Data Analyses      
The homoscedasticity (Levene’s test) and normality assumptions (Shapiro-Wilk test) for parametric terse were 
met. With that, analysis of variance (ANOVA) was employed for the mean comparison of induction and recovery 
time, followed by a posthoc test using Tukey’s test (p < 0.05). Regression analyses were used to determine the 
relationship between SL and induction time, and between SL and recovery time (p < 0.05). In the color container 
experiment, the data on survival rates, expressed as percentages were arc sine-transformed prior to analyses. 
Treatment means (mean ± SD) in every transport time were compared using ANOVA (P < 0.05). Data were 
presented as mean ± standard deviation (SD). All statistical analyses were performed with the SPSS v 17 and 
Paleontological Statistic v 3.0. 
3. Results 
3.1 Influence of Color Containers 
A summary of the survival rate of experimental fish is detailed in Table 2. Fish mortality occurred in the yellow 
and blue containers. The first mortality was observed in the yellow container, commenced at 6-h transport period, 
whereas the second mortality was recorded in the blue container at 12-h transport period. Nevertheless, no 
mortality happened in transparent, red, and black containers. Throughout, only two mortalities were recorded in 
the present study. Statistically, no significant difference among treatments was observed (P > 0.05). 
 
 
 



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Table 2. Data on the survival (%) of juvenile Leiopotherapon plumbeus packed in containers of different colors 
during the 12-h transport period 

Color of Containers 
Survival (%) 

2 h 4 h 6 h 8 h 10 h 12 h 
Transparent (Control) 100.00 100.00 100.00 100.00 100.00 100.00 

Red 100.00 100.00 100.00 100.00 100.00 100.00 
Yellow 100.00 100.00 93.33 93.33 93.33 93.33 
Black 100.00 100.00 100.00 100.00 100.00 100.00 
Blue 100.00 100.00 100.00 100.00 100.00 93.33 

 
3.2 Induction Time 
Different concentrations significantly affected the induction time, with a significant decrease in the induction 
period recorded (no more than 2 min) in T3 in each anesthetic solution (Table 3). The T1 of alcoholic drink had 
the longest mean induction time (9.73 ± 1.16 min). Nevertheless, the T3 of alcoholic drinks was able to anesthetize 
the fish individuals at 0.77 ± 0.14 min, considerably comparable to T3 of 2-phenoxyethanol (0.63 ± 0.12 min; Q 
= 1.11; p = 0.44), and T1 of MS-222 (0.74 ± 0.13 min; Q = 0.19; p = 0.89). Moreover, the mean induction time in 
T3 of benzocaine (1.96 ± 0.14 min) was not significantly different from T2 of the alcoholic drink (1.98 ± 0.30 min) 
(Q = 0.12; p = 0.93). Among all the treatments, the T3 of MS-222 solution had the most reduced mean induction 
time (0.21 ± 0.02 min).  
3.3 Recovery Time 
The recovery period was significantly varied among treatments in each anesthetic solution, with induced recovery 
time with increasing dosages (Table 4). The clove oil was found to cause longer mean recovery time relative to 
other anesthetic solutions (particularly for T1 and T3), whilst specimens exposed to 2-phenoxyethanol exhibited 
the fastest mean recovery time (1.02 ± 0.09 min). Fish specimens in all treatments of 2-phenoxyethanol and 
benzocaine had the fastest recovery time (no more than 3 min). However, two specimens died in T3 of the latter. 
Additionally, the mean recovery time for T1 of alcoholic drink (2.51 ± 0.50 min) was statistically homogeneous 
to T1 of MS-222 (2.68 ± 0.10 min), T2 (2.14 ± 0.28 min) and T3 (2.42 ± 0.32 min) of 2-phenoxyethanol, and T3 
of benzocaine (2.85 ± 0.27 min). Mean recovery time in T2 of alcoholic drinks (6.85 ± 0.97 min) showed 
considerable similar response to T2 of clove oil (6.03 ± 0.33 min; Q = 1.13; p = 0.43), and T3 of MS-222 (7.14 ± 
1.36 min; Q = 0.24; p = 0.86). 
 
Table 3. Induction rate (min) of different anesthetic solutions of varying concentrations for Leiopotherapon 
plumbeus (n = 200). 

Anesthetic 
Solutions 

Induction Time (min) F 
value p 

Control T1 T2 T3 

alcoholic drinks na 9.73 ± 1.16a 1.98 ± 0.30b 0.77 ± 0.14b 32.74 < 0.001

2-phenoxyethanol na 5.65 ± 1.21a 2.96 ± 0.64ab 0.63 ± 0.12b 13.97 < 0.001

clove oil na 1.26 ± 0.13a 1.11 ± 0.21a 0.37 ± 0.03b 28.19 < 0.001

MS-222 na 0.74 ± 0.13a 0.57 ± 0.12ab 0.21 ± 0.02b 12.08 < 0.001

benzocaine na 4.50 ± 0.29a 2.27 ± 0.27b 1.96 ± 0.14b 32.94 < 0.001

Treatments for alcoholic drink: T1 = 100 ml L-1, T2 = 200 ml L-1, T3 = 300 ml L-1 
Treatments for commercial anesthetic agents: T1 = 0.25 ml L-1, T2 = 0.5 ml L-1, T3 = 1.0 ml L-1 
For each anesthetic solution, means with the same superscript letter are not significantly different (Tukey post-
hoc tests); na = not applicable 
 
 



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Table 4. Recovery time (min) of Leiopotherapon plumbeus exposed to different anesthetic solutions of varying 
concentrations (n = 200) 

Anesthetic  
Solutions 

Recovery Time (min) F 
value p 

Control T1 T2 T3 

alcoholic drinks na 2.51 ± 0.50a 6.85 ± 0.97b 8.98 ± 0.84b 24.08 < 0.001 

2-phenoxyethanol na 1.02 ± 0.09a 2.14 ± 0.28b 2.43 ± 0.32b 14.18 < 0.001 

clove oil na 5.34 ± 0.27a 6.03 ± 0.33a 13.43 ± 0.91b 34.73 < 0.001 

MS-222 na 2.68 ± 0.10a 5.87 ± 0.91ab 7.14 ± 1.36b 10.72 < 0.001 

benzocaine na 1.97 ± 0.07a 1.92 ± 0.09a 2.85 ± 0.27b 5.27 < 0.001 

Treatments for alcoholic drink: T1 = 100 ml L-1, T2 = 200 ml L-1, T3 = 300 ml L-1 
Treatments for commercial anesthetics: T1 = 0.25 ml L-1, T2 = 0.5 ml L-1, T3 = 1.0 ml L-1 
For each anesthetic solution, means with the same superscript letter are not significantly different (Tukey post-
hoc tests); na = not applicable 
 
3.4 Correlation of Fish Length with Induction and Recovery Time 
Correlation analysis indicated that the induction and recovery time is size-influenced, with smaller specimens 
anesthetized faster than the larger ones (Table 5). This observation was significantly evident in T1 of all anesthetic 
solutions, as well as in T2 of alcoholic drinks. The size of the fish did not affect the induction time when the 
concentration of commercial anesthetics was 0.5 ml L-1 and above. No significant correlation between size and 
recovery time was observed in T1 of alcoholic drinks and MS-222, including T2 and T3 of all anesthetic solutions 
(Table 5).  
 
Table 5. Relationship of standard length (mm) with induction and recovery times in each treatment. Significant 
relationships are set in bold. 

Anesthetic Solutions  
T1 T2  T3 

R2 p R2 p  R2 p 

Alcoholic Drink 
SL vs Induction 0.78 0.001 0.85 0.002  0.34 0.329
SL vs Recovery 0.48 0.157 0.28 0.583  0.46 0.181

         

2-Phenoxyethanol 
SL vs Induction 0.71 0.019 0.59 0.073  0.52 0.126
SL vs Recovery 0.76 0.010 0.61 0.060  0.54 0.110

         

Clove Oil 
SL vs Induction 0.69 0.026 0.59 0.072  0.55 0.102
SL vs Recovery 0.66 0.038 0.18 0.622  0.19 0.597

         

MS-222 
SL vs Induction 0.71 0.020 0.55 0.098  0.61 0.062
SL vs Recovery 0.62 0.057 0.20 0.573  0.41 0.267

         

Benzocaine 
SL vs Induction 0.73 0.018 0.34 0.337  0.62 0.056
SL vs Recovery 0.72 0.020 0.30 0.392  0.45 0.268

Treatments for alcoholic drink: T1 = 100 ml L-1, T2 = 200 ml L-1, T3 = 300 ml L-1 
Treatments for commercial anesthetic agents: T1 = 0.25 ml L-1, T2 = 0.5 ml L-1, T3 = 1.0 ml L-1 
 
 



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4. Discussion 
The present study demonstrated that the color background did not affect the survival of L. plumbeus juveniles 
during the 6-h and 12-h transport times. Previous studies demonstrated that background color affects the 
performance of various aquatic species (Luchiari and Freire 2004; Ninwichian et al. 2018; Manliclic et al. 2019). 
In the works of Manliclic and his colleagues (2018), survival of Nile tilapia juveniles improved when exposed to 
the blue color container in 24-h conditioning time before transport. In the present study, no significant difference 
was observed in the mortality rate of L. plumbeus juveniles. It is comparable to the result of Manliclic et al. (2018), 
where there are no significant differences in four different color backgrounds during 6-h and 12-h transport time.  
The induction times decreased significantly as concentrations increased in 2-phenoxyethanol, clove oil, MS-222, 
and benzocaine. The results are similar to the previous studies suggesting an inversed relationship between 
induction time and concentration of anesthetic agents in teleost fishes (Pawar et al. 2011; Yildiz et al. 2013; Varkey 
and Sajeevan 2014; Kucuk and Coba 2016; Ogretmen et al. 2016; Bolasina et al. 2017; Park 2019). It is also 
apparent that the alcoholic drinks have comparable effect in several dosages of commercial anesthetic agents 
The study provides evidence that alcoholic beverages containing 40% ethanol can be used as an effective 
anesthetizing agent for L. plumbeus. The presence of an active component, ethanol is known to inhibit or depress 
the central nervous system activity of many mammalian species (Banerjee 2014). In most fish farms in the 
Philippines, alcoholic drinks are usually used as an anesthetic agent for aquaculture fish (e.g., Clarias gariepinus). 
In the absence of commercial anesthetics, alcoholic drinks can be practical options to facilitate the smooth handling 
of fish during induced spawning. The present finding thus opens further studies as to the anesthetic effect of 
alcoholic drinks in L. plumbeus during actual breeding and transport.  
Recovery times increased gradually with increasing concentrations of all the anesthetic agents. Prolonged recovery 
with increased anesthetic dosage had been reported in seahorse (Pawar et al. 2011), rainbow trout (Yildiz et al. 
2013), redline torpedo fish (Varkey and Sajeevan 2014), goldfish (Kucuk and Coba 2016), shabbout fish 
(Ogretmen et al. 2016), guppy (Bolasina et al. 2017), catfish (Park 2019). However, several studies have 
documented a decreasing recovery time with an increase in the concentration of clove oil and 2-phenoxyethanol 
for European sea bass (Dicentrachus labrax) and gilthead seabream (Sparus aurata) (Mylonas et al. 2005). 
The mechanisms of fish in recovery times during anesthetizing seem to be complex. The observed delayed 
recovery may be attributed to its persistence on the gill surface (Sladky et al. 2001; King et al. 2005). Moreover, 
the difference in the respective recovery times is highly affected by species, size, physiological status, and 
environmental conditions (Ross and Ross 1999). Different factors, i.e., biological and/or environmental factors 
affect the efficacy of anesthetics in fish. According to Coyle et al. (2004), the efficacy of anesthetic drugs depends 
on the gill area to body weight ratio, metabolic rate, and temperature related. Moreso, larger fish has a slower 
absorption rate of anesthetic drugs compared to smaller fish because of the smaller gill area surface relative to 
body mass for drug diffusion (Popovic et al. 2012). Our observation is in agreement with the work of Park (2019), 
i.e., small-sized fish were more easily anesthetized and recovered more rapidly from anesthesia than large-sized 
fish. Despite that, there are other reports that found that larger individuals had quicker recovery times than smaller 
ones (Woody et al. 2002; Fernandes et al. 2017). It is noteworthy to mention that all recovered and survived fish 
individuals in the two experiments were returned to hatchery facility for future research initiatives.   
5. Conclusion and Recommendations 
This preliminary study demonstrated that the color of containers had no profound effect on the survival of juvenile 
L. plumbeus, although this may be attributed to low stocking density and short transport period. For further 
investigation, it is suggested to increase the number of experimental fish per container and increase the transport 
time under the ambient setting. 
The present study demonstrated the anesthetic efficacy of alcoholic drinks and four commercial anesthetic agents 
in L. plumbeus. Higher dosages, viz 200 ml L-1 to 300 ml L-1 of alcoholic drinks were found to be effective, and 
are comparable to the induction response of other commercial anesthetics. The results imply cost-effective use of 
anesthetic during induced spawning and transportation of this terapontid. The results of the study provide a 
practical method for minimizing stress in fish handling, which is vital in aquaculture and conservation endeavors 
for this terapontid, and possibly in other diminutive native fish species. 
The study can be replicated in induced spawning and transportation under ambient set-up. Fish euthanization using 
experimental anesthetic agents is also open for further investigation. Likewise, investigation of the dose-response 
in biological status (reproductive state, sizes, and sexes), hematological profile, and cortisol levels will contribute 
more to the total efficacy of anesthetics in experimental L. plumbeus. 



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Acknowledgment 
The study is funded by the Research and Development Office of Bataan Peninsula State University. Special 
appreciation is accorded to the student-researchers, viz Ma. Angelica Swin, Christian Jarrel Perez, Richard San 
Diego, Giselle Refuerzo, Erica Shane Enriquez, and Lea Raya for lending their helping hand and hard works, and 
to the anonymous reviewers for the comments and suggestions. 
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    /HRV (Za stvaranje Adobe PDF dokumenata najpogodnijih za visokokvalitetni ispis prije tiskanja koristite ove postavke.  Stvoreni PDF dokumenti mogu se otvoriti Acrobat i Adobe Reader 5.0 i kasnijim verzijama.)
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    /NLD (Gebruik deze instellingen om Adobe PDF-documenten te maken die zijn geoptimaliseerd voor prepress-afdrukken van hoge kwaliteit. De gemaakte PDF-documenten kunnen worden geopend met Acrobat en Adobe Reader 5.0 en hoger.)
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    /ENU (Use these settings to create Adobe PDF documents best suited for high-quality prepress printing.  Created PDF documents can be opened with Acrobat and Adobe Reader 5.0 and later.)
  >>
  /Namespace [
    (Adobe)
    (Common)
    (1.0)
  ]
  /OtherNamespaces [
    <<
      /AsReaderSpreads false
      /CropImagesToFrames true
      /ErrorControl /WarnAndContinue
      /FlattenerIgnoreSpreadOverrides false
      /IncludeGuidesGrids false
      /IncludeNonPrinting false
      /IncludeSlug false
      /Namespace [
        (Adobe)
        (InDesign)
        (4.0)
      ]
      /OmitPlacedBitmaps false
      /OmitPlacedEPS false
      /OmitPlacedPDF false
      /SimulateOverprint /Legacy
    >>
    <<
      /AddBleedMarks false
      /AddColorBars false
      /AddCropMarks false
      /AddPageInfo false
      /AddRegMarks false
      /ConvertColors /ConvertToCMYK
      /DestinationProfileName ()
      /DestinationProfileSelector /DocumentCMYK
      /Downsample16BitImages true
      /FlattenerPreset <<
        /PresetSelector /MediumResolution
      >>
      /FormElements false
      /GenerateStructure false
      /IncludeBookmarks false
      /IncludeHyperlinks false
      /IncludeInteractive false
      /IncludeLayers false
      /IncludeProfiles false
      /MultimediaHandling /UseObjectSettings
      /Namespace [
        (Adobe)
        (CreativeSuite)
        (2.0)
      ]
      /PDFXOutputIntentProfileSelector /DocumentCMYK
      /PreserveEditing true
      /UntaggedCMYKHandling /LeaveUntagged
      /UntaggedRGBHandling /UseDocumentProfile
      /UseDocumentBleed false
    >>
  ]
>> setdistillerparams
<<
  /HWResolution [2400 2400]
  /PageSize [612.000 792.000]
>> setpagedevice

