Investigation of the aquaculture potential of an Australian freshwater prawn, Macrobrachium spinipes

171 Pages·2015·1.56 MB·English

by  Malwine Lober

ResearchOnline@JCU

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Lober, Malwine (2015) Investigation of the aquaculture
potential of an Australian freshwater prawn,
Macrobrachium spinipes (Schenkel, 1902), with emphasis
on spawning induction, larval and nursery culture. MSc
thesis, James Cook University.

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i

Investigation of the aquaculture potential of an Australian
freshwater prawn, Macrobrachium spinipes (Schenkel, 1902),
with emphasis on spawning induction, larval and nursery
culture

Thesis submitted by

Malwine Lober, BSc, Akld, NZ, GradDip Res Meth, JCU, Qld

in March, 2015

for the degree of Master of Science
in the College of Marine and Environmental Sciences
James Cook University
ii

STATEMENT ON THE CONTRIBUTION OF OTHERS

I wish to thank the following for the assistance towards my studies at James Cook
University. Firstly, the opportunity provided by the John Allwright Scholarship of the
Australian Centre for International Agricultural Research (ACIAR) to undertake
postgraduate studies, for that I am greatly thankful. I thank my supervisor A/Prof. Chaoshu
Zeng and Prof. Paul Southgate for guidance throughout my research. Particular thanks to
Dr. Chaoshu Zeng for his patience and guidance in editing of this thesis and publication.
The staff of the Department of Primary Industries and Fisheries (DPI&F), Walkamin,
Cairns for collection of broodstock and the use of the facility to hold and maintain
broodstock, without the continued availability of brooders this research would have been
very challenging. Financial support for my research was provided by JCU through the
annual IRA and Grant Research Scheme (GRS). Finally, thanks to Professor Peter Mather
of the Queensland University of Technology, for genetic identification of prawn samples.

Declaration on Ethics

This research presented and reported in this thesis was conducted in compliance with the
National Health and Medical Research Council (NHMRC) Australian Code of Practice for
the Care and Use of Animals for Scientific Purposes, 7th Edition, 2004 and the Qld Animal
Care and Protection Act, 2001. The proposed research study received animal ethics
approval from the JCU Animal ethics Committee Approval Number # A1046
iii

ACKNOWLEDGEMENTS

I would like to thank Evizel Seymour, Peter Graham and staff of the Freshwater Fisheries
and Aquaculture Centre, Queensland Department of Primary Industries and Fisheries
(QDPI&F), Walkamin for maintenance and supply of broodstock. Also, I thank Mr. John
Morrison and staff at the Marine and Research Facility Unit (MARFU), JCU for setting up
the laboratories for experimentation and maintenance of the systems throughout my
research. Mr. Brendan Fry for collection of broodstock prawns. Dr. Chan Lee, ACIAR, for
his guidance and encouragement to do postgraduate studies. I would also like to thank Mr.
Haruo Tsuji and Ms. Noordiyana Mat Norrdin, for donating your valuable time to help out
in the laboratory and in the field. Lastly, I thank the invaluable moral and financial support
of my family during my studies at JCU.
iv

Abstract
Modern aquaculture of freshwater prawns began in 1960’s after the life cycle of
Macrobrachium rosenbergii was first closed. The industry has since expanded rapidly with
global annual production reaching 443,959 tonnes in 2009. The Australian native
freshwater prawn species formerly known as M. rosenbergii was recently renamed as
Macrobrachium spinipes. The past attempts to commercially culture the native Australian
freshwater prawn has never taken off citing various problems. In view that new molecular
evidence showed native Australian freshwater prawn encompasses four lineages, this study
investigated broodstock management, larval and nursery culture techniques for the lineage
II M. spinipes sourced from tropical north Queensland.

Spawn induction of M. spinipes outside its natural breeding season (November to March)
was investigated by temperature and photoperiod manipulation. Mature females (15) were
kept individually under controlled water temperatures of 24, 27 and 30 °C with photoperiod
adjusted to 14.5 h light: 9.5 h dark while the control was under ambient conditions. The
experiment ran from May to August for 84 days and throughout the experimental duration,
no spawning was recorded in any treatments. Despite ambient temperature dropping to as
low as 12 °C for the control, no mortality was recorded in all treatments except a single
death found for 30 °C treatment and common moults occurred in all treatments even during
the coldest period in the control. Hence M. spinipes showed high tolerance to low water
temperature and potential for grow-out culture under cooler climates.
v

The effects of microalgae Nannochloropsis sp. addition and concentration (‘green water’)
on larval survival, development and growth of newly hatched larvae of M. spinipes were
investigated under the condition of no algae addition (‘clear water’) and four
Nannochloropsis concentrations of 2.5, 6.25, 12.5 and 25 x 105 cells ml-1. Survival to PL at
the two higher algae concentrations of 12.5 and 25 x 105 cells ml-1 (70.8 % and 63.3 %,
respectively) were significantly higher (P<0.05) than the two lower concentrations and the
‘clear water’ treatments (26.7 %, 35.0 % and 30.0 %), respectively. Meanwhile, the fastest
mean development to PL (30.6 days) was obtained at the highest algal density, which was
14 days shorter than that of the ‘clear water’ treatment (44.3 days). Mean dry weights of
newly settled PL’s in the two high algal density treatments were also significantly heavier
(P<0.05) than in the lowest algal density and the ‘clear water’ treatments. The results
clearly showed that in comparison with the ‘clear water’ method, ‘green water’ culture by
adding Nannochloropsis sp. potentially could substantially improve larval performance of
M. spinipes, however such beneficial effects only became significant when the
concentration of algae added reached a threshold level.

A further experiment was conducted to evaluate the effects of addition of higher microalgae
concentrations as compared to Artemia enrichment on larval growth, survival and fatty acid
composition of M. spinipes larvae and their interactions. The newly hatched larvae were
cultured with no addition of algae (‘clear water’) and Nannochloropsis sp. added (‘green
water’) at two concentrations of 2.5 (low) and 10 x 106 cells ml-1(high) and fed with either
enriched or non-enriched Artemia: i.e.1) ‘clearwater’ + enriched Artemia; 2) ‘clearwater’ +
non-enriched Artemia; 3) 2.5 x 106 cells ml-1 + enriched Artemia; 4) 2.5 x 106 cells ml-1 +
non-enriched Artemia; 5) 10 x106 cells ml-1 + enriched Artemia and 6) 10 x106 cells ml-1 +
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non-enriched Artemia. The results showed that under ‘clear water’ culture, larvae fed non-
enriched Artemia resulted in total mortality while Artemia enrichment improved survival to
18 %; at the low algal density (2.5x106 cells ml-1), larval survival also improved with
Artemia enrichment (55.8 vs. 80 %). However, survival was very similar with and without
Artemia enrichment at high algal density of 10x106 cells ml-1. ‘Clear water’ culture resulted
in poorer larval survival, while on the other hand it showed that Artemia enrichment
improved survival mainly under ‘clear water’ or low algal density condition. Meanwhile,
increased algal density and Artemia enrichment were shown to significantly improve larval
development and growth, respectively. The shortest mean development times to PL (20.5
and 21.5 days) were from both 10 x106 cells ml-1 treatments with enriched treatment having
the shortest 20.5 days while non-enriched treatment at slightly longer 21.5 days.
Meanwhile, all ‘greenwater’ treatments showed advanced ‘Larval Stage Index’ (LSI)
compared with the ‘clear water’ treatments. In contrast, dry weights and total lengths of PL
from enrichment treatments were heavier or larger. It is concluded that Artemia enrichment
was most beneficial for larval culture when ‘clearwater’ method was used or microalgae
was added at low concentrations in ‘green water’ culture.

Newly settled postlarvae (PL) of M. spinipes were stocked under three densities of 500
(low), 1000 (intermediate), 2000 PL m-3 (high) and with or without shelter, respectively to
investigate the effects of stocking density and shelter as well as their potential interactions
on the nursery culture of M. spinipes. Statistical analysis detected no significant interaction
(P>0.05) between the two factors of stocking density and shelter within the range tested,
the main effects were therefore analysed separately. With the survival results of the two
treatments under a same density (i.e. with and without shelter) pooled, it showed that the
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low density (500 PL m-3) produced significantly higher mean survival (84.8 %) than the
high density treatment (2000 PLm-3) (57.4 %) (P<0.05). The intermediate density (1000
PLm-3) generated an intermediate result (74.4 %), which was significantly different
(P<0.05) from both the high and low density treatments. At the end of culture, the high
stocking density also produced significantly heavier juveniles (wet weight: 144.24 ± 0.01
mg ind-1), significantly higher final biomass (13.45 ± 1.39 g) and increase in biomass (8.87
± 1.28 g) than that of the low density treatment (wet weight: 117.99 ± 0.003 mg ind-1; final
biomass: 4.69 ± 0.16 g and increase in biomass: 2.95 ± 0.15 g). Meanwhile, the addition of
shelter produced significantly heavier juvenile wet weight (134.3 ± 0.01 vs. 123.61 ± 0.003
mg ind-1) and higher final biomass (11.76 ± 1.84 vs. 8.79 ± 1.12 g). The results suggest that
stocking density of 2000 PL m-3 improved final production while 500 PL m-3 improved
survival. On the other hand, under the same stocking densities, shelter addition enhanced
juvenile wet weight and final biomass.

In summary, the results of the present study suggest that the Australian native M. spinipes
is a freshwater Macrobrachium species that shares many biological and aquaculture
similarities with M. rosenbergii but also has some major differences. As it is native to
Australia, M. spinipes has high potential for aquaculture in the country, therefore should be
investigated further for its potential commercial farming in Australia and also possibly
overseas.
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Table of Contents
Statement on the Contribution of Others........................................................................... ii
Acknowledgements........................................................................................................... iii
Abstract............................................................................................................................. iv
Table of Contents.............................................................................................................. viii
List of Tables..................................................................................................................... xi
List of Figures................................................................................................................... xiii
List of Plates...................................................................................................................... xv
Chapter 1: Introduction.................................................................................................. 1
1.1 Freshwater Prawn Aquaculture in Australia and Objectives of This Study............ 1
1.2 Aquaculture History and Global Production of Macrobrachium rosenbergii…… 2
1.3 Nomenclature and Natural Distribution................................................................... 4
1.4 General Biology of Macrobrachium rosenbergii.................................................... 11
1.4.1 Life cycle and habitat......................................................................................... 11
1.4.2 Diets and appendages for feeding...................................................................... 12
1.4.3 Reproductive biology......................................................................................... 13
1.4.3.1 Sexual dimorphism and morphotypes of males................................................. 13
1.4.3.2 Reproduction seasonality................................................................................... 15
1.4.3.3 Mating, spawning, egg hatching and larval development.................................. 15
1.5 M. rosenbergii Aquaculture Practices.................................................................... 20
1.5.1 Broodstock management.................................................................................... 20
1.5.2 Larval culture..................................................................................................... 23
1.5.2.1 Larval feeds........................................................................................................ 23
1.5.2.2 ‘Greenwater’ vs ‘Clearwater’ larval culture method........................................ 24
1.5.2.3 Stocking density, duration of larval culture and other culture conditions....... 25
1.5.3 Nursery............................................................................................................... 28
1.5.4 Growout.............................................................................................................. 32

Chapter 2: Attempt to induce out-of-season spawning of Macrobrachium spinipes
(Schenkel, 1902) by temperature and photoperiod manipulation.............................. 34
2.1 Abstract.................................................................................................................. 34
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2.2 Introduction............................................................................................................ 35
2.3 Materials and Methods............................................................................................ 38
2.3.1 Source of female prawns.................................................................................... 38
2.3.2 Experimental design and general procedure...................................................... 38
2.3.3 Data Analysis..................................................................................................... 40
2.4 Results.................................................................................................................... 42
2.4.1 Water quality parameters................................................................................... 42
2.4.2 Survival.............................................................................................................. 42
2.4.3 Moulting and intermoult duration...................................................................... 42
2.4.4 Growth................................................................................................................ 43
2.5 Discussion............................................................................................................... 53

Chapter 3: Effect of microalgae concentration on larval survival, development
and growth of an Australian giant freshwater prawn Macrobrachium
spinipes (Schenkel, 1902)................................................................................................ 56
3.1 Abstract................................................................................................................... 56
3.2 Introduction............................................................................................................. 58
3.3 Materials and Methods ........................................................................................... 60
3.3.1 Source of broodstock and larvae ........................................................................60
3.3.2 Experimental design and setup........................................................................... 61
3.3.3 Data Analysis...................................................................................................... 64
3.4 Results ..................................................................................................................... 64
3.4.1 Larval survival.................................................................................................... 64
3.4.2 Larval development............................................................................................ 65
3.4.3 Postlarval dry weight and carapace length....................................................... 66
3.5 Discussion ............................................................................................................... 73

Chapter 4: The effect of Artemia enrichment and green water culture on larval
survival, development and postlarval fatty acid composition of a native Australian
giant freshwater prawn Macrobrachium spinipes (Schenkel, 1902) .......................... 80
4.1 Abstract .................................................................................................................. 80
4.2 Introduction ............................................................................................................ 82
4.3 Materials and Methods............................................................................................ 85