July | August Feature heading: Essential Oils Feature title: The benefits of oregano essential oil for aquaculture production International Aquafeed is published five times a year by Perendale Publishers Ltd of the United Kingdom. All data is published in good faith, based on information received, and while every care is taken to prevent inaccuracies, the publishers accept no liability for any errors or omissions or for the consequences of action taken on the basis of information published. ©Copyright 2009 Perendale Publishers Ltd. All rights reserved. No part of this publication may be reproduced in any form or by any means without prior permission of the copyright owner. Printed by Perendale Publishers Ltd. ISSN: 1464-0058
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Essential Oil
Essential Oil Oregano essential oil, with its renowned antimicrobial and antioxidant properties, is a good potential alternative to growth promoters in this respect.
The benefits of
oregano essential oil for aquaculture production
by Justin Tan Yu-Wen, DVM
T
he use of phytobiotic feed additives in aquaculture has gained a steady increase since restrictions were implemented on the use of in-feed antibiotic growth promoters in the livestock industry. Due to increasing demand for high quality protein sources, we have witnessed a rapid growth of the fin-fish and shrimp farming industries, where typical production involves high-density stocking rates at both hatchery and grow-out stages. Due to its highly intensive nature, fin-fish and shrimp farming today are plagued by diseases affecting productivity and survival rates. Various strategies have been devised and implemented to improve growth rates while preventing and controlling incidences of disease and pathogen levels. Besides the development of new vaccines and improved water management techniques, the use of feed additives such as phytobiotics and essential oils are now widely accepted for their ability to improve growth rates through notable beneficial effects on intestinal health.
The passing of the era of AGPs
stances that showed promise and offered potential as replacements. The intestinal microbiota of aqua species should not be ignored in relation to their performance. Although fin-fish and shrimp have only marginal nutritional benefits from intestinal microbiota compared to other terricolous animals such as ruminant and non-ruminant herbivores, the microbiota can adversely affect the resilience of the host if not properly controlled. It has now become evident that controlling the microbiota could positively influence growth rates and immune status.
Antibiotic growth promoters (AGPs) have played a fundamental role in aquaculture production throughout the last 50 years. These are thought to act on the intestinal microbiota, leading to improved animal growth and performance. However, we have seen a trend in the last decade, where an extensive search for alternatives to these feed additives has taken place. This paradigm shift is chiefly due to public concerns related to their residues in meat and the subsequent occurrence of antibiotic-resistant bacteria. Feed additive manufacturers and feed producers have invested a considerable amount of time, money and resources to investigate and Figure 1: Minimum Inhibitory Concentrations of various conduct extenbotanical essential oils against 3 types of common sive research on microorganisms (Source: Hammer et al., 1999) various ingredients and sub16 | InternatIonal AquAFeed | July-august 09
Time to go natural Meriden Animal Health Limited (UK) produces a unique feed additive called Orego-Stim® Aquatract, which contains oregano essential oil. Various studies have shown that OregoStim® Aquatract can improve growth performance and feed conversion efficiency, while greatly reducing the occurrence of disease. This ultimately leads to an increase in profitability compared to conventional aquaculture rearing systems. The matrix of active components of pure oregano essential oil found in OregoStim® Aquatract have been shown to be highly effective against both gram-positive and gram-negative pathogenic bacteria (Stiles et al., 1995; Sivropoulou et al., 1996) but has little or no effect on beneficial bacteria that make up the majority of the intestinal microbiota (Di Pasqua et al., 2005; Jintasataporn et al., 2007). Oregano essential oil was compared with many other known botanical essential oils and its antibacterial properties were tested against three common species of pathogenic bacteria. In this study, oregano essential oil was found to be the most effective in the inhibition of these pathogens (see Figure 1).
The use of OregoStim® Aquatract in fin-fish production A commercial trial was conducted by a reputable international aquafeed producer from Denmark that produces rainbow trout feed. Rainbow trout (Oncorhynchus mykiss) were fed with either standard control feed or a similar feed containing Orego-Stim® Aquatract at 550ppm. The length of the trial was 49 days and the fish were reared in a Danish trout farm using a flow-through system, with water temperatures of 9.4°C. Results revealed that trout fed with feed containing Orego-Stim® Aquatract showed improvements in average weight gain by 9.4 percent (see Figure 2), total weight gain by 14.9 percent and feed efficiency by 3.2 percent (see Figure 3). This ultimately resulted in a Return-OnInvestment (ROI) ratio of 8.6 : 1, which meant that for every €1 they spent on OregoStim® Aquatract, they gained an additional
profit of €8.60 in return on top of usual profits, due to the additional increase in liveweight gain. Jintasataporn et al. (2007a) conducted a 12-week trial using Orego-Stim® Aquatract, demonstrating its positive effects on the growth performance of hybrid catfish (Clarias macrocephalus x Clarias gariepinus). Results showed that fish fed with Orego-Stim® Aquatract had a higher body weight compared to both positive and negative control groups, with a significant increase in body weight by 10.7% and 11.7% respectively (see Figure 4). A more efficient feed conversion ratio was also observed in the OregoStim® Aquatract group, which was 6.38% and 4.96% lower than both positive and negative control groups respectively (see Figure 5).” Orego-Stim® Aquatract also revealed contrasting antimicrobial activity against pathogenic and beneficial bacteria (Jintasataporn et al., 2007a). Contrary to a common misconception that Orego-Stim® may also inhibit the growth of beneficial bacteria, this study proved otherwise. The following results show that Orego-Stim® Aquatract was able to increase the numbers of lactic acid producing bacteria such as Lactobacillus spp. in the gastrointestinal microbiota by up to 62.5% compared to the negative control group, whilst reducing populations of pathogens such as Aeromonas spp., a bacterial species that commonly affects freshwater fish (see Figure 6).
Figure 2: Growth performance: Average weight gain (g) of rainbow trout (Source: Meriden Animal Health Ltd. Research & Technical Report, 2007)
Figure 3: Growth performance: Feed conversion ratio of rainbow trout (Source: Meriden Animal Health Ltd. Research & Technical Report, 2007)
Figure 4: Growth performance: Weight (g/fish) (Source: Jintasataporn et al., 2007a)
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Figure 5: Growth performance: Feed Conversion Ratio (Source: Jintasataporn et al., 2007a)
Essential Oil
Essential Oil The use of Orego-Stim® Aquatract in shrimp production
Figure 6: Effect of Orego-Stim® on Bacterial Count (Source: Jintasataporn et al., 2007a)
Figure 7: Shrimp production: Feed Conversion Ratio (Source: Merchan, 2005)
Figure 8: Shrimp production: Survivability (%) (Source: Merchan, 2005)
Figure 9: Shrimp immunity: Prophenoloxidase Activity (Source: Jintasataporn et al., 2007b)
Improvements in feed conversion efficiency and survival rates were also reported in shrimps. A four-month commercial trial conducted in Ecuador demonstrated improvements in feed conversion by 19.8 percent (see Figure 7) and survival rates by 6.5 percent (see Figure 8) by adding Orego-Stim® Aquatract to the diet of shrimps, compared to the control group, which were fed the same diet but without the inclusion of Orego-Stim® Aquatract. The results clearly indicated how Orego-Stim® Aquatract could improve shrimp farming production systems compared to conventional rearing systems (Merchan, 2005). Jintasataporn et al. (2007b) also conducted a three-month trial in white shrimps (Litopenaeus vannamei) and discovered that by using Orego-Stim® Aquatract at 1000ppm, the prophenoloxidase activity was increased by 170 percent (or 2.7 times) and 69 percent (or 1.7 times) higher than that of the control and enrofloxacin groups respectively (see Figure 9). The prophenoloxidaseactivating system is a major innate defense mechanism of invertebrates and prophenoloxidase activity is a good indicator of shrimp immunity. Further evaluation was carried out to determine the relationship between the increased prophenoloxidase activity and disease resistance in
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shrimp. The shrimps were subjected to virulent Vibrio harveyi challenge after the initial three-month growth promotion trial. Results showed that Orego-Stim® Aquatract had a protective effect on the shrimp, resulting in a significantly lower mortality rate of 36.67 percent in comparison with the negative control group and the enrofloxacin group, which had mortality rates of 56.67 percent and 53.33 percent respectively (see Figure 10). Similar and corresponding to the aforementioned catfish trial, higher levels of oregano essential oil, albeit being a potent antibacterial agent, did not exert detrimental effects on lactic acid producing bacteria such as Lactobacillus spp. although on the other hand, bacterial load and Vibrio spp. counts were markedly reduced in the Orego-Stim® Aquatract 2000ppm group compared to the negative control and enrofloxacin groups (see Figure 11). This finding clearly explains the increased survivability rate in shrimps with the use of the higher inclusion rates of Orego-Stim® Aquatract during critical periods and times of stress and disease challenge.
Conclusion The use of Orego-Stim® Aquatract has been proven to be effective in enhancing growth performance, improving the immune status and preventing the occurrence of diseases in both fin-fish and shrimps. However, it must be noted that not all phytobiotics offer the same benefits or efficacy in terms of results and productivity. Due to the differences that can be found in the levels and ratios of active chemical constituents of the many, diverse plant extracts, the variance in bioavailability and bioactivity of these active components may result in differing performances from those reported (Jin et al., 2008). As there are more than 30 different active components that are naturally found in pure oregano essential oil, it is highly likely that these other minor constituents play an important role in the efficacy of Orego-Stim® Aquatract, as there is evidence that the antimicrobial
essential oils from Mediterranean Apiaceae, Verbenaceae and Lamiaceae against foodborne pathogens and spoilage bacteria,’ Annals of Microbiol., Vol. 55(2), pp. 139-143.
Figure 10: Shrimp immunity: Mortality rate upon disease challenge with Vibrio harveyi (Source: Jintasataporn et al., 2007b)
Figure 11: Shrimp immunity: Bacterial Count (cfu log10/g) (Source: Jintasataporn et. al., 2007b)
activity of whole, unadulterated oregano essential oil is greater than the additive effect of its major antimicrobial components (Lattaoui and Tantaoui-Elaraki, 1994). It is also crucial to note that commercially available ‘nature identical’ versions of oregano essential oil require a degree of purity of not less than 99.5 percent in order to comply with food legislation (Ariza-Nieto, 2005). Any degree of purity that is less than this would not be deemed suitable for entry into the human food chain. Therefore, it is crucial for nutritionists and feed producers to consider these facts when choosing from a range of phytobiotic or essential oil feed additives for improving aquaculture production, to ensure compliance with standard food safety regulations on a global scale.
References: Ariza-Nieto, C.J., 2005, ‘Evaluation of Oregano (Origanum vulgare) Essential Oil in Swine Production System’, Doctoral Thesis, University of Minnesota, United States of America. Di Pasqua, R., De Feo, V., Villani, F. & Mauriello, G., 2005. ‘In vitro antimicrobial activity of
Hammer, K.A., Carson, C.F. & Riley, T.V., 1999, ‘Antimicrobial activity of essential oils and other plant extracts,’ J. Appl. Microb., Vol. 86, pp. 985-990. Jin, L.Z. & Lan, G.Q., 2008, ‘The research application of plant extract feed additives in swine production’, Chinese Swine Production Magazine, Vol. 3.
1996, J. Agrc. Food Chem., Vol. 44, pp. 12021205. Stiles, J.C., Sparks, W. & Ronzio, R. A., 1995, J. Appl. Nutr., Vol. 47, pp. 96-102. Strand, M.R., 2008, ‘The insect cellular innate response’. Insect Science, Vol. 15, pp. 1-14.
Fin Fish and Shrimp Improves FCR Increases bodyweight gain Boosts immunity Increases survival rates Reduces feed costs
= Higher returns
Jintasataporn, O., Areechon, N., Ritchai, N. & Tonedial, C., 2007a, ‘The effect of Orego-Stim® in Catfish’, Unpublished Data, Kasetsart University, Thailand. Jintasataporn, O., Areechon, N., Ritchai & N., Tonedial, C., 2007b, ‘The effect of OregoStim® in White Shrimp Litopenaeus vannamei’, Unpublished Data, Kasetsart University, Thailand. Lattaoui, N., & A. TantaouiElaraki, 1994. ‘Individual and combined antibacterial activity of the main constituents of three essential oils.’ EPPOS, Vol. 3, pp. 13-19. Merchan, R., 2005, ‘Effects of Orego-Stim in Shrimp Production: Ecuador Study’, Unpublished Data, Merchan & Fontana Company, Ecuador. Meriden Animal Health Limited Research & Technical Report, 2007, Unpublished data, United Kingdom Sivropoulou, A., Papanikolaou, E., Nikolaou, C., Kokkini, S., Lanara, T. & Arsenakis, M.,
100% Natural Additive for inclusion in the feed
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