Multienzymatic capacity of cultivable intestinal bacteria from captive Litopenaeus vannamei (Boone, 1931) shrimp reared in green water
- 52 Downloads
Equilibrium between the gastrointestinal bacterial population and the environment is a critical factor for the health of captive aquatic animals. The bacterial enzymes are fundamental for proper nutrition and pathogen resistance in shrimp. Therefore, enzymatic profiles reveal essential characteristics for the selection of probiotic strains that can improve animal development. In this work, we analysed shrimp from a green water system where infectious myonecrosis virus (IMNV) was present. We isolated transient and intestine resident bacterial populations, characterising eight functional groups through different culture media. To identify each isolated bacteria, we used sequences from regions V6–V8 of the 16S rRNA. To determine viral load of shrimp samples, we used real-time PCR. The number of colony forming units (CFU) was similar between IMNV-infected and IMNV-uninfected shrimps. The growth of transient bacteria was higher than the growth of resident. In general, lipolytic bacteria presented higher frequency and genus diversity than the other functional groups. All groups showed higher frequency among transitory bacteria, except the amylolytic functional group, which was more frequent among the resident. We found two major orders of cultivable bacteria, Vibrionales and Bacillales. The genus Vibrio was predominant among the Vibrionales, while Staphylococcus and Bacillus were the most frequent among the Bacillales. Recorded Vibrionales and Bacillales included pathogenic and beneficial species of high importance for aquaculture. The results presented here will serve as a basis for improving the nutritional and health conditions of Litopenaeus vannamei in green water farming systems.
KeywordsLitopenaeus vannamei Gut bacteria Green water IMNV Enzymatic group Probiotics
Infectious myonecrosis virus
White spot syndrome virus
Colony forming unit
Cultivable heterotrophic bacteria
We would like to thank the team at Laboratory of Environmental Microbiology and Fish (LAMAP, UFC) for the help in carrying out all the microbiological analysis processes. The authors kindly thank Ms. Kellie Johns, from James Cook University, for English revisions.
This work was funded by CAPES (CIMAR 2000/2014). J. Forte was supported by a CAPES PhD grant from CIMAR 2000/2014.
Compliance with ethical standards
All shrimp were collected and maintained in accordance with ethical standards in animal research.
Conflict of interest
The authors declare that they have no conflict of interest.
- Adel M, El-Sayed AFM, Yeganeh S et al (2016) Effect of potential probiotic Lactococcus lactis Subsp. lactis on growth performance, intestinal microbiota, digestive enzyme activities, and disease resistance of Litopenaeus vannamei. Probiotics Antimicrob Proteins 9:1–7. https://doi.org/10.1007/s12602-016-9235-9 CrossRefGoogle Scholar
- Andrade TPD, Srisuvan T, Tang KFJ, Lightner DV (2007) Real-time reverse transcription polymerase chain reaction assay using TaqMan probe for detection and quantification of infectious myonecrosis virus (IMNV). Aquaculture 264:9–15. https://doi.org/10.1016/j.aquaculture.2006.11.030 CrossRefGoogle Scholar
- APHA – American Public Health Association (2000) Standard Methods for the Examination of Water and Wasterwater. Greenberg AE, Clesceri LS, Eaton AD (20th eds) APHA/AWWA/WEF, p 1–10. https://nla.gov.au/anbd.bib-an22987801. Accessed 19 Oct 2017
- Cardona E, Gueguen Y, Magré K, Lorgeoux B, Piquemal D, Pierrat F, Noguier F, Saulnier D (2016) Bacterial community characterization of water and intestine of the shrimp Litopenaeus stylirostris in a biofloc system. BMC Microbiol 16:1–9. https://doi.org/10.1186/s12866-016-0770-z CrossRefGoogle Scholar
- Façanha FN, Oliveira-Neto AR, Figueiredo-Silva C, Nunes AJP (2016) Effect of shrimp stocking density and graded levels of dietary methionine over the growth performance of Litopenaeus vannamei reared in a green-water system. Aquaculture 463:16–21. https://doi.org/10.1016/J.AQUACULTURE.2016.05.024 CrossRefGoogle Scholar
- Hostins B, Lara G, Decamp O, Cesar DE, Wasielesky W (2017) Efficacy and variations in bacterial density in the gut of Litopenaeus vannameireared in a BFT system and in clear water supplemented with a commercial probiotic mixture. Aquaculture 480:58–64. https://doi.org/10.1016/j.aquaculture.2017.07.036 CrossRefGoogle Scholar
- Kearse M, Moir R, Wilson A, Stones-Havas S, Cheung M, Sturrock S, Buxton S, Cooper A, Markowitz S, Duran C, Thierer T, Ashton B, Meintjes P, Drummond A (2012) Geneious basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics 28:1647–1649. https://doi.org/10.1093/bioinformatics/bts199 CrossRefPubMedPubMedCentralGoogle Scholar
- Luis-Villaseñor IE, Voltolina D, Audelo-Naranjo JM, Pacheco-Marges MR, Herrera-Espericueta VE, Romero-Beltrán E (2015) Effects of biofloc promotion on water quality, growth, biomass yield and heterotrophic community in Litopenaeus vannamei (Boone, 1931) experimental intensive culture. Ital J Anim Sci 14:332–337. https://doi.org/10.4081/ijas.2015.3726 CrossRefGoogle Scholar
- Maturin L, Peeler JT (2001) Aerobic Plate Count. In: Food and Drug Administration (FDA), Bacteriological Analytical Manual Online, 8th Chapter 3. Edition, Silver Spring, Berlin. http://www.fda.gov/food/foodscienceresearch/laboratorymethods/ucm063346.htm. Accessed 16 Oct 2017
- Ngo HT, Nguyen TTN, Nguyen QM, Tran AV, Do HTV, Nguyen AH, Phan TN, Nguyen ATV (2016) Screening of pigmented Bacillus aquimaris SH6 from the intestinal tracts of shrimp to develop a novel feed supplement for shrimp. J Appl Environ Microbiol 121:1357–1372. https://doi.org/10.1111/jam.13274 CrossRefGoogle Scholar
- Nunes AJP, Sá MVC, Sabry-Neto H (2011) Growth performance of the white shrimp, Litopenaeus vannamei, fed on practical diets with increasing levels of the Antarctic krill meal, Euphausia superba, reared in clear- versus green-water culture tanks. Aquac Nutr 17:e511–e520. https://doi.org/10.1111/j.1365-2095.2010.00791.x CrossRefGoogle Scholar
- Phuoc LH, Corteel M, Thanh NC, Nauwynck H, Pensaert M, Alday-Sanz V, den Broeck WV, Sorgeloos P, Bossier P (2009) Effect of dose and challenge routes of Vibrio spp. on co-infection with white spot syndrome virus in Penaeus vannamei. Aquaculture 290:61–68. https://doi.org/10.1016/j.aquaculture.2009.02.004 CrossRefGoogle Scholar
- Raghul SS, Bhat SG (2011) Seasonal variation in the hydrolytic exo-enzyme profile of Vibrio sp. associated with the marine benthic environment of South India. Indian J Geomarine Sci 40:826–833Google Scholar
- Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, New YorkGoogle Scholar
- Sha Y, Wang L, Liu M, Jiang K, Xin F, Wang B (2016) Effects of lactic acid bacteria and the corresponding supernatant on the survival, growth performance, immune response and disease resistance of Litopenaeus vannamei. Aquaculture 452:28–36. https://doi.org/10.1016/j.aquaculture.2015.10.014 CrossRefGoogle Scholar
- Sierra G (1957) A simple method for the detection of lipolytic activity of micro-organisms and some observations on the influence of the contact between cells and fatty substrates. Antonie Van Leeuwenhoek 23:15–22. https://link.springer.com/article/10.1007/BF02545855 CrossRefGoogle Scholar
- Vargas-Albores F, Porchas-Cornejo MA, Martínez-Porchas M, Villalpando-Canchola E, Gollas-Galván T, Martínez-Córdova LR (2017) Bacterial biota of shrimp intestine is significantly modified by the use of a probiotic mixture: a high throughput sequencing approach. Helgol Mar Res 71:5. https://doi.org/10.1186/s10152-017-0485-z CrossRefGoogle Scholar
- West PA, Colwell RR (1984) Identification and classification of Vibrionaceae - an overview. In Vibrios in the Environment, RR Colwell (Editor). John Wiley, New York, p 285–363Google Scholar