|Year : 2019 | Volume
| Issue : 1 | Page : 8-15
Development of a bait carrier material for apple snail (Pomacea maculata) based on its feed preferences using snail attractant tracking device
Guruswamy Prabhakaran, Subhash Janardhan Bhore, Manikam Ravichandran
Department of Biotechnology, Faculty of Applied Sciences, AIMST University, Semeling, 08100 Bedong, Kedah, Malaysia
|Date of Web Publication||4-Feb-2019|
Department of Biotechnology, Faculty of Applied Sciences, AIMST University, Bedong-Semeling Road, Semeling, 08100 Bedong, Kedah
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Background: Synthetic molluscicides are routinely used in the control of apple snails (Pomacea spp of Ampullariidae family), the invasive rice pest. Molluscicidal bait is a strategic combination of food and toxicant to attract the target pests. However, baits are found to be feed deterrent due to snails' own feed preferences in a paddy field. There is no specific feed formulation exclusively developed as the base carrier material for the development of molluscicidal bait for the apple snail (Pomacea maculata Perry). Further, there are limitations in evaluating the efficacy of a snail feed/bait formulation based on its attractiveness, palatability, and directional movement of snails with the existing experimental devices. Therefore, this study was aimed to develop an attractive and appetent feed formulation as a bait carrier material and also to design and develop a snail attractant tracking device (SATD) for evaluating the feed formulation precisely. Materials and Methods: Snails were collected from the snail-infested rice field located in Kedah, Malaysia. A glass aquarium tank of size 60 cm × 30 cm × 30 cm was physically partitioned to form four linear channels (tracks) using 5-mm thickness transparent perflex plastic sheets to develop SATD and snail feed formulations were evaluated with the help of it. Results: This study resulted in the development and evaluation of attractant feed formulations prepared with a combination of different types of carbohydrates and protein hydrolysates. Analysis of variance showed significant differences (P < 0.05) in the attraction and consumption of various types of feed formulations by the snails. Conclusion: The rice flour supplemented with 5% sugar, 15% wheat flour, and 0.5% fish meal showed a maximum attraction and effective consumption by snails (1.48 g/3 h) than the other tested feeds.
Keywords: Bait attractiveness, feed formulations, golden apple snail, molluscicides, rice
|How to cite this article:|
Prabhakaran G, Bhore SJ, Ravichandran M. Development of a bait carrier material for apple snail (Pomacea maculata) based on its feed preferences using snail attractant tracking device. J Nat Sc Biol Med 2019;10:8-15
|How to cite this URL:|
Prabhakaran G, Bhore SJ, Ravichandran M. Development of a bait carrier material for apple snail (Pomacea maculata) based on its feed preferences using snail attractant tracking device. J Nat Sc Biol Med [serial online] 2019 [cited 2019 Jun 26];10:8-15. Available from: http://www.jnsbm.org/text.asp?2019/10/1/8/251495
| Introduction|| |
Rice is the staple food and an essential crop grown worldwide, with Asia being the largest producer and consumer. Apple snails (Pomacea spp. of Ampullariidae family) are the major invasive rice pest. The golden apple snail (GAS) Pomacea canaliculata (Lamarck, 1822) has voracious appetite for the young rice seedlings of both transplanted and direct-seeded rice. GAS infestations resulted in huge economic loss of $1.47 billion per annum in rice production in Southeast Asian countries. In Peninsular Malaysia, Pomacea maculata (Perry, 1810) is more abundant than P. canaliculata in the major paddy growing regions., Further, due to a higher fecundity rate P. maculata poses a serious threat than P. canaliculata to aquatic ecosystems., Hence, the present study was focused on P. maculata. As a part of the Integrated Pest Management, the application of niclosamide, a synthetic molluscicide is the mainstay of GAS control programs. The cost of synthetic molluscicides, their toxicity to nontarget organisms, and persistence in the environment have led to the development of molluscicidal bait formulations., Molluscicide bait is target specific with minimal adverse effect on the nontarget organisms and environment than the application of conventional molluscicides directly in the water. Plant-derived molluscicides have long been explored as effective alternatives to synthetic chemical counterparts for control of apple snails.,,,
Snails use olfaction in foraging and the food materials serve as chemical cues responsible for the principal of attraction. Besides, the chemical cues play predominant roles in snails' prey detection, alarm responses, species recognition, and reproduction.,,, In aquatic system, the various feeds containing carbohydrates and amino acids were attracted by gastropods. Notably, starch was found to be an effective attractant for the Lymnaea acuminata snails., Molluscicidal baits are formulated with snail preferred food materials such as wheat bran or pasta as the carrier for the active ingredient and the principal attractant., The apple snails are primarily macrophytophagous and extremely polyphagous on a broad range of diets and the rice field provides a variety of feed choices It was further evidenced that rice was least preferred by GAS compared to rice weeds., With regard to feed preferences, both the P. canaliculata and P. maculata were established as generalist compared to other Pomacea species in their predominant habitats. Studies also revealed the complexity in feeding habits and feed preferences at different stages of these snails. In paddy field, commercial molluscicidal baits are mostly found to be feed deterrent due to the high concentration of active ingredients and snails' own feed preferences. Consequently, there exists an uncertainty in terms of attractiveness and its consumption of commercial molluscicide baits in a given ecosystem. Hence, it demands the development of appetent bait which can distinctly attract the rice pest snails. According to Ogbu et al., the concept of snails' feed preference should encompass two major features of the feed; such as its attractiveness (measured by choice tests) and palatability (measured by comparing the quantity of the different feedstuffs ingested) for preparing effective bait formulations with molluscicides. Furthermore, Howlett et al. emphasized that the efficacy of bait should be determined by the directional movement of the slugs toward the target and the number of slugs affected by ingestion of bait. Paradoxically, most commercial molluscicidal baits contain 2%–6% active ingredients, and hence, instead of bait, it serves as a feed deterrent to snails.,
Various laboratory experimental devices have been developed to evaluate the attractiveness of snails to sex pheromones/feed/bait formulations. Singh and Singh designed a bioassay chamber with a round glass aquarium tank and evaluated a bait formulation for its attractiveness to a freshwater snail (Lymnaea acuminata). Whereas, Baker et al. designed a Y-tube olfactometer and assessed the attraction of grain pest snails (Cernuella virgata, Theba pisana, and Cochlicella acuta) toward odors elicited from 22 potential foodstuffs. Alternatively, Ogbu et al. used a rectangle wooden cage to ascertain the feeding preference of two African giant land snails (Achatina achatina and Archachatina marginata). In another study, Takeichi et al. used a T-maze device made of acryl board and reported the attraction of adult male P. canaliculata to female sex pheromones and the juveniles' preference to the food-conditioned water. However, in all these devices, there are certain limitations in evaluating the attractiveness of a feed/bait material. Essentially, in these experimental devices without any physical barriers (to serve as guiding channels), the movement of snails would be distance oriented. Distance oriented in the sense that the snails can travel in a zigzag manner to reach the target food. Hence, it is difficult to monitor the directional movement and measurement of actual distance travelled by a snail accurately in an experimental condition. Therefore, it necessitates the design and development of snail attractant tracking device (SATD) for the development and evaluation of feed formulations accurately for Pomacea spp. So far, no feed formulation was exclusively developed and evaluated as the base carrier material for the development of molluscicidal bait for P. maculata. Hence, the present study was initiated with the aim to develop a bait carrier material for P. maculata based on its feed preferences. Further, to design and develop a SATD for evaluation of the feed formulations precisely based on its attractiveness and palatability.
| Materials and Methods|| |
To collect the snails, no specific permissions were required. The snail species used in this study is not an endangered or a protected species in Malaysia. In Malaysia, the collection of snails for study purpose is not regulated under any act. Therefore, to initiate this study, the review and approval of the study protocol by the suitable ethics committee and or institutional biosafety committee was not a precondition.
Collection of apple snails
Snails with a shell length of 10 mm to 30 mm were collected from the snail-infested rice field located in Semeling, Kedah, Malaysia. The snails were identified based on the conchological characters as reported., The identification of species was confirmed by the authority at District Agriculture Office, Sungai Petani, Kedah, Malaysia. The snail's shell length was measured with a vernier caliper for ascertaining the shell length of a snail. The snails were acclimated in 20 l laboratory aquarium tank (length [l] 60 cm × width [w] 30 cm × height [h] 30 cm) containing dechlorinated tap water at room temperature (25°C ± 2°C). The snails were fed with fresh lettuce (Lactuca sativa) ad libitum., The juvenile snails (20–30 mm of shell length) actively moving and feeding on lettuce were considered healthy and used in experiments.
Design of snail attractant tracking device
A clean rectangular glass aquarium tank (l × w × h – 60 cm × 30 cm × 30 cm) was physically partitioned with the transparent perflex plastic sheets to form four longitudinal lanes/tracks in the bottom of the tank for the movement of snails from one end to another end of the tank [Figure 1]. Three partition sheets of dimension (l × w × h – 60 cm × 7.5 cm × 7.5 cm) were used for the formation of tracks. The width of each track was 7.5 cm. In each partitioning sheet, at both ends, a rectangular slit of 15 cm × 4 cm was made to facilitate the movement of snails. Besides, two support frames were fixed horizontally to hold the partition sheets in the tank. The height of the partitioning sheet not only restricted the snails crossing the lanes but also channelized their movement. The aquarium tank was filled with 3 l of dechlorinated tap water to a height of 6 cm. A feed formulation prepared either as granules or a cake was kept in a Petri plate and placed at one end of the tank. At another end, 10 snails of 20–30 mm size were placed in a row. The number of snails that were attracted and came in contact with the feed and the time taken by each snail to travel the distance (60 cm) in a track was monitored at every 5 min until 1 h or more at room temperature (25°C ± 2°C). The time taken by a snail to reach the feed formulation and the quantity of feed consumed by the snail were used as a measure of a feed's attractiveness and preference.
|Figure 1: Design of snail attractant tracking device. (a) Design of tracks with partition frames and support frames made from perflex sheets for placement in the aquarium tank and (b) Aquarium tank with tracks for snails' movement for the study of attraction of snails by feed|
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Development of a bait carrier material for apple snail
The feed formulations prepared with different types of food grade starches such as rice, wheat, corn, and tapioca flours individually or in combination with crude sources of amino acids (yeast extract powder [Himedia], commercial grade fish meal, and chicken feather hydrolysate) were evaluated for its attractiveness and effective consumption by snails. The flow chart of experiments for the development of a bait carrier material for P. maculata is depicted in [Figure 2].
|Figure 2: Flowchart of experiments for the development of a bait carrier material for apple snail (Pomacea maculata). C: Corn starch; R: Rice flour; S: Sugar; T: Tapioca starch; W: Wheat flour|
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Identification of a suitable carbohydrate as a feed material for snail
To identify a suitable attractive base feed material for apple snails, four different carbohydrates were evaluated. Food grade rice flour, wheat flour, cornstarch, and tapioca starch were individually used for the batter preparation. The batter was prepared by adding 10 g of rice flour with 10 ml of distilled water and mixed thoroughly to obtain the dough consistency. The dough was then rolled manually into granules of diameter 2–5 cm and allowed to dry at 60°C for 12 h. Similarly, the wheat granules were prepared using wheat flour (10 g in 10 ml water). The cornstarch and tapioca starch (10 g of each) were used as such. The snails were not fed with lettuce for 24 h before experiments. The snails were transferred to another holding tank containing dechlorinated tap water and allowed to starve for additional 24 h to remove any residual chemical cues present at the onset of the experiment. At the start of the experiment, 10 snails were placed in a row at one end of the SATD filled with dechlorinated tap water and allowed to acclimate for 5 min. Subsequently, at another end, the test feed in ungranulated or granulated or in cake form was placed in a Petri plate. The feed was evaluated for its attractiveness and preference by recording the time taken by snails to reach the feed and the quantity of feed consumed at room temperature (25°C ± 2°C). The experiments were conducted during morning hours from 8 am to 12 noon. Experiments were conducted in triplicates to evaluate the different feed preparations for its attractiveness and palatability.
The rate of consumption of different carbohydrates by snails
To investigate the consumption of different feeds by apple snails, the batter material was prepared as cakes instead of granules. Rice flour (10 g) was added to 100 ml of boiling water and heated in a micro oven until the thick glutinous slurry was formed. The slurry was spread in Petri plates to form a layer of cake and allowed to cool at room temperature and used. Likewise, the wheat, corn, and tapioca slurries were prepared by boiling 15 g wheat, 8 g corn starch, and 12 g tapioca starch individually in 100 ml water and spread in Petri plates. The different feed cakes kept in Petri plates were individually fed to 30 snails in SATD for a period of 3 h to determine the rate of consumption of feed. Feed preference was assessed based on the consumption of a feed formulation (3 h/g) by the snails. The feed consumption was determined by the change in Petri plate weight from initial to final weight using a Mettler Toledo AL 104 balance. Experiments were triplicated with 30 snails per se t.
The attractiveness of carbohydrate feed supplemented with sugar
Sugar at different concentrations (5%, 10%, and 15%) were supplemented to the test carbohydrates and evaluated. The batter was prepared and used for the preparation of granules as described earlier. The rice flour supplemented with three different concentrations of sugar (9.5 g rice flour + 0.5 g sugar, 9.0 g rice flour + 1.0 g sugar, and 8.5 g rice flour + 1.5 g sugar) was tested individually for its attractiveness to snails in the SATD. Similarly, the wheat flour, cornstarch, and tapioca starch supplemented with sugar were prepared individually and evaluated for its attractiveness. Experiments were performed in triplicate with 10 snails per se t. The difference in attractiveness for each combination of feed was tested using one-way ANOVA followed by Tukey post hoc test in IBM-SPSS (version 22) statistical program (IBM Corp., Armonk, NY, USA) at the 0.05% significant level.
Evaluation of the combination of carbohydrates as a feed
The rice flour supplemented with 5% sugar was found to be effective in terms of its attractiveness and preference by the snails from the earlier experiments. This feed formulation was supplemented with wheat flour at three different concentrations (10%, 15%, and 20%) and the combined carbohydrates feed was evaluated for its attractiveness. The batter was prepared by adding 8.5 g rice flour + 0.5 g sugar + 1 g wheat flour, 8.0 g rice flour + 0.5 g sugar + 1.5 g wheat flour, and 7.5 g rice flour + 0.5 g sugar + 2 g wheat flour, respectively, in 10 ml of distilled water and mixed thoroughly to obtain dough consistency. The dough was then granulated as described earlier. The attractiveness of the combined carbohydrates feeds was individually evaluated in the SATD.
Evaluation of the combined carbohydrates feed supplemented with different protein hydrolysates
A combined carbohydrates feed (rice flour supplemented with 5% sugar and 15% wheat) was selected based on its attractiveness and preference by the snails from the earlier experiments. To this combined carbohydrates feed, the yeast extract, chicken feather hydrolysate, and fish meal were incorporated individually at the rate of 0.5% and evaluated for its attractiveness. Incorporation of fish meal enhanced the feed attractiveness. Therefore, to optimize its concentration in the combined feed experiments were conducted with three different concentrations (0.5%, 1%, and 2%) of fish meal. The rate of attraction of the snails to the different feeds with respect to time was recorded individually.
Consumption of different feed formulations by snails
Granular feed formulations made of rice flour, wheat flour, and sugar and fish meal are shown [Figure 3]a. The consumption of various feed formulations by snails was comparatively evaluated. The feed granules such as (a) rice flour; (b) rice flour with 5% sugar; (c) rice flour, 5% sugar, and 15% wheat; (d) rice flour, 5% sugar, 15% wheat, and 0.5% fishmeal; and (e) rice flour, 5% sugar, 15% wheat, and 1.0% fish meal were individually feed to 30 snails in the SATD for a period of 3 h to determine the rate of consumption of a feed.
|Figure 3: (a) Granular feed formulations made of rice flour, wheat flour, and sugar and fish meal. (b) Evaluation of different carbohydrates as the snail attractant feed. All the values are mean ± standard error of the mean (n = 10). **P < 0.01 compare with wheat; #P < 0.05 compare with corn. (c) Consumption of different carbohydrates as feed by the snails. ***P < 0.001 compared with rice. 3.2. The attractiveness of carbohydrate feed supplemented with sugar, and (d) Evaluation of rice flour supplemented with sugar as snail attractant feed. *P < 0.05 compared with rice|
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The mean ± standard error of the mean values was calculated for each group. The difference in attractiveness for each combination of feed was tested using one-way ANOVA followed by Tukey post hoc test in SPSS (version 22) at the 0.05% significant level.
| Results|| |
Identification of suitable carbohydrate as the snail attractant feed
The results presented in [Figure 3]b indicate the trend of the attractiveness of different carbohydrates to the snails. Among the flours, wheat, corn, and rice were found to be effective attractants to snails than the tapioca. Compared to rice (16 min), wheat flour attracted the snails in short duration (12 min). Although the wheat flour attracted the snails, the consumption rate was the highest for rice (700 mg/3 h) followed by wheat (230 mg/3 h) as recorded [Figure 3]c. The consumption of the corn was less (108 mg/3 h) and the tapioca was not at all consumed by the snails. The attractiveness of a feed is mostly depending on its chemical composition, while the palatability is influenced by texture and softness of the feed., Therefore, rice and wheat flour were identified as the base carbohydrate materials for the development of snail attractant feed.
Addition of 5% sugar to the rice flour had attracted the snails more than the other concentrations (10% and 15%) of sugar [Figure 3]d. The addition of 5% sugar had reduced the attraction time by 5 min compared to the rice flour alone (control). On the contrary, the addition of sugar to wheat, corn, and tapioca did not bring a notable reduction in time [Figure 4]a. Among the flours tested, rice flour supplemented with 5% sugar was found effective in attracting the snails. Rice flour supplemented with 5% sugar was found more attractive to snails than the rice flour with higher sugar concentration (10% and 15%). The snails took shorter time (9-14 min) to reach the rice flour supplemented with 5% sugar. Whereas, snails took longer time (22-25 min) to reach the rice supplemented with 15% sugar.
|Figure 4: (a) Evaluation of different carbohydrates supplemented with sugar as snail attractant feed. Values are mean ± standard error of the mean (n = 10). *P < 0.05 compared with rice. (b) Evaluation of the combination of carbohydrates as snail attractant feed. (c) Evaluation of combined carbohydrates feed supplemented with different protein hydrolysates. RSW: Rice + 5% sugar and 15% wheat. **P < 0.01 compare with RSW + Yeast extract 0.5%; ##P < 0.001 compare with RSW + Chicken feather hydrolysate 0.5%, and (d) Consumption of different carbohydrates feed supplemented with fishmeal by snails. **P < 0.01; ***P < 0.001 compare with rice|
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Evaluation of the combination of carbohydrates as a feed
The rice flour supplemented with 5% sugar was identified as the base material for the feed formulation. Next to rice, the wheat flour was effectively attracted by snails and hence studies were conducted to find out whether a combination of rice and wheat flour would further enhance its attractiveness to snails. Addition of 15% wheat flour to the base material (rice + 5% sugar) had enhanced the attractiveness of the feed than the control [Figure 4]b. In that combination, snails took 11 min to reach the feed settings, whereas in control, it took 13 min. Hence, the combined carbohydrates feed (rice + 5% sugar and 15% wheat) was selected for the further development.
The addition of 0.5% fish meal to the combined carbohydrates feed had attracted the snails within 9 min compared to the control base feed (13 min) as shown in [Figure 4]c. The addition of chicken feather meal and yeast extract to the combined carbohydrates feed did not enhance its attractiveness to the snails.
Comparative analysis of different feed formulations for its attractiveness and consumption by snails
The comparative analysis of the attractiveness and consumption of various feed formulations by snails is presented in [Figure 4]b,[Figure 4]c,[Figure 4]d. The results indicate that the feed formulation consists of rice flour supplemented with 5% sugar, 15% wheat flour, and 0.5% fish meal was effectively consumed (1.48 g/3 h) by the snails compared to other feed formulations tested. The formulation attracted the snails faster by 3–4 min compared to other combinations of feed and hence, it is identified as an effective bait carrier material for P. maculata.
| Discussion|| |
Snails use olfactory and gustatory cues in identifying and ingesting their food. Different feed formulations were evaluated in a specially designed SATD which facilitated the development of appetent feed formulation for apple snail (P. maculata). The comparative advantages of designed SATD over the existing experimental devices for the study of the attraction of snails by food/sex pheromones are depicted in [Table 1].
|Table 1: Comparative analysis of existing experimental devices for the study of the attraction of snails by food/sex pheromones|
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In the designed SATD, the guiding channels regulated the movement of snails in a unidirectional way to reach the target food. The design can be extrapolated at a higher scale to 4–8 ft rectangular aquarium tanks to study the attractiveness of a feed/bait formulation. Moreover, the design considerations are feasible to find out the optimum distance for placement/dispersal of bait formulations in the field conditions. The device helped to find out the velocity of snails while they are attracted toward food. Higher the velocity of snails, higher is the attractiveness of the food/bait.
In the present study, the wheat flour attracted the snails in short duration (12 min) than rice flour (16 min). This observation supports the wide usage of wheat bran as an attractive carrier material for molluscicidal bait formulations.,, Among the various carbohydrates tested, the rice flour was consumed at a higher rate (700 mg/3 h) than the wheat flour (230 mg/3 h) by snails. The higher consumption rate of rice flour may attribute to its physical and chemical properties besides starch being the major form of storage polysaccharide in most plants. Similarly, the binary combination of sucrose and starch (10 mM of each) was found to be an effective attractant for the L. acuminata snails as reported., Notably, our results indicated that the addition of higher concentration of sugar (5%) to the rice flour has significantly enhanced its attractiveness to P. maculata. Further, the results also revealed that tapioca starch and corn starch were found to be inappetent feed, both individually or in combination with sugar for P. maculata and it may be due to species-specific nutrient requirement., In addition to carbohydrates, amino acids (proline and serine) were also found to be effective attractants for the aquatic snail L. acuminata and Indoplanorbis exustus., Our results clearly indicate that incorporation of fish meal (0.5%) to the base carbohydrates feed has enhanced its attractiveness to P. maculata and it might be due to the amino acids content in the fish meal. The results are in conformity with the findings of Kumar et al. who reported that the combination of attractant carbohydrates and amino acids in bait formulation showed the highest attraction of L. acuminata snails. The authors have developed a synergistic polyherbal molluscicidal formulation with the ethanolic extracts of Nerium indicum, Nicotiana tabacum, Piper nigrum, and Azadirachta indica against P. maculate. The polyherbal formulation will be incorporated in the feed formulation for development of a bait formulation toward the effective management of P. maculata in a rice field.
| Conclusion|| |
In the present study, using the SATD, the specific feed formulation was exclusively developed based on its attractiveness and palatability to P. maculata. Eventually, this base feed formulation would serve as a bait carrier material for incorporating molluscicidal compounds. In the SATD, the guiding channels facilitated in tracking the movement at different experimental times to determine the velocity of snails when they were attracted toward food, precisely. The design concept of SATD is simple and can be also used to investigate study the attractiveness and effectiveness of feed or bait formulation with other pest snails and slugs.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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[Figure 1], [Figure 2], [Figure 3], [Figure 4]