Manipulating nutrient and environment to support healthy food chain

1. Ibnu Sahidhir
Senior Aquaculture Engineer
Brackishwater Aquaculture Development Center Ujung Batee, Aceh
1
Microbial food web
Maintaining Healthy Shrimp Pondforforforfor
A Novel Approach
Microbial
Manipulating
Food Web

2. Main Questions
1. What is the significances of microbes in the pond ?
2. What is the microbial food web ?
3. How to manipulate bacterial food web for supporting shrimp health ?
4. How to manipulate phytoplankton food web for supporting shrimp
health ?
5. What is the right pond condition for better microbial food web to
flourish ?

3. CONSIDERING
THE ENERGY
Bacteria and phytoplankton
as basic of microbial food chain
Photosynthesis
Protozoa
Decomposition
Metazoa
pH
pH

4. I
The Significances of Microbes in the Pond

5. Microbial ServicesMicrobial ServicesMicrobial ServicesMicrobial Services
in Ponds
Detoxifying
wastes
• Absorbing and converting
NH3, H2S, CO2, malodor
Preventing
diseases
• Maintain
biodiversity
Providing
nutrition
• Vitamins,
immunostimulant
etc

6. 0 50 100 150
Fertilizer
Soil quality
Broodstock feed
Antibiotics and disinfectant
Weather and health management
Seed
Larval feed
Growout feed
Alternative feed
Probiotic and bioremediation
My Personal Website Survey of Aquaculture Problem
10/28/2019 6
(www.sahidhir.com 2014)
Rank 1st : Probiotic
Number of People

7. Probiotics 15,200 entry
Prebiotics 4,430 entry
Synbiotics 1,600 entry
Postbiotics 33 entry
microbes
microbial food
microbes and
its food
Microbial metabolites
product

8. II
The Microbial Food Web

9. Microbial Community Succession from Organic Matter
Nannoflagellates
Organicpollution

10. Light intensity of tropical sun light 4000 kkal/m2/day

11. Microbial Community Succession from Inorganic Matter
Phytoplankton
Free ciliates
Attached ciliates
Copepod
Rotifer
Inorganicpollution

12. Attached
Dispersed
Flocculated
Settled
Distribution of Microbes
Foam and scum

13. (Wang et al., 2014)

14. METHODS
Finding
stuck in
food
chain
Preparing
healthy
basic of the
pyramides
Helping
the
grazer
Controlling
the
environment

15. Day 1st:
Heterotrophic bacteria decompose organic matter

16. Day 7th
Microalgae grows after inorganic mineral accumulated

17. Filamentous bacteria as a skeleton of floc

18. Young Floc

19. Pennales Diatom in Floc

20. Attached Ciliata in Old Floc

21. Ciliates-Rotifer-Gastrotrich as Floc Grazer

22. Bakteri 0,1-1 um
Protozoa 0,5-20 um
Rotifer 100-300 um
Organisme bersel banyak yang paling sederhana

23. Gastrotrichs

24. Crustacean and Nematodes (300-1500 um)

25. Microbial
Food Web
in ERAS
Organic matter Solid dissolved
Bacteria
Organic
Anorganic Phytoplankton
Micro
Meso
Protozoa
Nanno
Micro
Macro
Free swimming
Crawling
Attached
Contractile
Non-contractile
swimming
floc
attached
Rotifers
Nematods
Copepods
Insects
(mosq larvae,
chironomus)
Ciliata
(70%)
Melosira, Navicula
Bio-agitation and coprophagy
Predation
CO2 in
CO2 Out
Biomass decreases
Biomass increases

26. Biomass
Pool
CO2 in
CO2 Out
Biomass decreases
Biomass increases
Simplifying the Concept

27. Indoor Outdoor
Floc Volume in Outdoor is Higher than Indoor

28. 3
10
0,00
2,00
4,00
6,00
8,00
10,00
12,00
14,00
weight
(gram)
Indoor
Outdoor

29. Nitrite-N
Total Ammonia-N
Nitrate-N
0
5
10
15
20
25
30
35
0
0,05
0,1
0,15
0,2
0,25
0,3
0 8 12 15 19 26 28
Days
Nitrate-N
(mg/l)
TAN dan Nitrite-N
(mg/l)
A Complete Ammonia Oxidation Pattern (COMMAMOX by Nitrospira sp)
When microbial food web is running well, No Worries about Nutrient Cycles

30. Effect of ciliated protozoa on the activated sludge quality
Effluent analysis Without ciliates With ciliates
Bacteria (millions/ml) 106-160 1-9
BOD (mg/l) 53-70 7-24
Organic nitrogen (mg/l) 14-21 7-10
Suspended solids (mg/l) 86-118 26-34

31. MANIPULATING THE MICROBIAL FOOD WEB
Finding
stuck in
food
chain
Preparing
healthy
basic of the
pyramides
Helping
the
grazer
Controlling
the
environment

32. III
Manipulating Bacterial Food Web

33. Microbial food chain become stuck because of
filamentous microbes

34. 23 Clade of Vibrio
Deadly AHPND Vibrio
transfer the plasmid
to Orientalis clade (V. punensis)
(Clade: Sawabe et al, 2013)
(AHPND: Restrepo, 2018)

35. (Dworkin et al, 2006: The prokaryotes VI)Physiological Profile of Vibrios

36. Natural microbial community
Cell
Metabolites
Stimulate protozoa
Stimulate floc forming bacteria
Reduce total bacteria
Reduce
pathogenic bacteria
probiotic
postbiotic
Organic acid and ethanol
Heterotrophic nannoflagellates
• induce polyhydroxyalkanoates
• Polyphosphate accumulating bacteria
Postbiotic Strategy of Heterofermentative Microbes

37. Classical RABAL (Yeast and Lactic Acid Bacteria) of Heterofermentative Natural Strain

38. SelfSelfSelfSelf----purification of waterpurification of waterpurification of waterpurification of water
4 days water maturation
Lavilla-Pitogo CR, Albright LJ, Paner MG (1998)

39. Effect of ciliated protozoa on the activated sludge quality
Effluent analysis Without ciliates With ciliates
Bacteria (millions/ml) 106-160 1-9
BOD (mg/l) 53-70 7-24
Organic nitrogen (mg/l) 14-21 7-10
Suspended solids (mg/l) 86-118 26-34

40. Homofermentative (hexose) Heterofermentative (hexose & pentose)
Lactobacillus acidophilus, L casei, L. plantarum,
L. kefiranofaciens, L. kefirgranum
Lactobacillus fermentum, L. brevis, L. kimchii, L.
Sakei, L.
Lactococcus lactis, Lactococcus plantarum Leuconostoc kimchii, L. Lactis, L. mesenteroides
Paralactobacillus selangorensis Carnobacterium divergens
Pediococcus acidilactici Oenococcus oeni
Vagococcus fluvialis Weisella kimchii, W. koreensis
Lactic Acid Bacteria
(Jay, 2005; Dworkin et al, 2006: The prokaryotes IV)

42. Polyphosphate Accumulating Microbes after PHA Formation
(Yuan, 2012)

43. (Janssen, 2012)

44. Microbial Floc:
a Rain Forest in the Dessert of Shrimp Faming
• Site of biodiversity
• Nitrifiers shelter
• P accumulators shelter
• Benthic diatom shelter
• Maintain safe decomposition

45. IV
Manipulating Microalgal Food Web

46. Silicate in seawater 8 ppm
Si (20% silicate) 1,6 ppm
N (5x Si) 8 ppm
Feed protein 36%
N protein 16%
N secretion 70%
Total N waste 4%
Feed 198 ppm
Shrimp size (150 ind/m3) 1,3 gram
Shrimp size (300 ind/m3) 0,7 gram
Predicting when Si used up
Postbiotics prevents microalgal collapse by slowing population growth

47. Big size of microalgae makes microbial food chain become stuck

48. • Silicon is limiting nutrient
• Diatom tolerance to defficiency is 20%, thin frustule
• After 10 days 40% of organic Si is mineralized
• Size of Diatom influence mineralization of organic Si

50. Asexual Sexual
Sexual mode on when resources is depleted
Binary fission Oogonium Spermatogonium Engagement
ZigoteFertilization
Cyclotella menegheniana

51. Protozoa grazing bacteria in scum

52. • Exopolymeric substances (green)
(carbohydrates)
• Abundant when nutrient (NP) limited
EPS of Spirulina

53. Lack of protozoa in anaerobic sediment

54. Filamentous Bacteria in Upper Layer Anaerobic Sediment

56. Sulfate 2700 ppm
Sulfide potential (33% of sulfate) 900 ppm
Sulfide toxicity 0,2 ppm
Killing area 4500 m3
Sulfate = 2,7 ppt
Macro Anion Disaster in Anaerobic Environment

57. Before and After
Periphyton Feeder
Comes
Tilapia grazing
prevent mirobial food
chain become stuck

58. V
Providing Excellent Environment
for Microbial Food Web to Flourish

59. A Simplified Process in Traditional Pond:
Nothing can be Controlled

60. Thinking Like
Bioreactor
• Control DO (>4 ppm) and pH (7,8-8,6)
• Harvest the microbes regularly by
siphoning

61. Ca/Mg/K = 1/3,5/1 = 2 ppt
Macro Cation

62. Current Water Management: Horisontal vs Vertical

63. Current and aeration make different storyCurrent and aeration make different storyCurrent and aeration make different storyCurrent and aeration make different story
No water exchange
for 2 months
TAN 0,08 ppm
Nitrite 0,004 ppm
Nitrate 11,9 ppm
Phosphate 0,13 ppm
TVC 230 cfu/ml

64. 82%
86%
92%
96%
80%
82%
84%
86%
88%
90%
92%
94%
96%
98%
100%
0 50 100 150
Dose of Postbiotics (ppm)
Survival Rate in Nursery
10/28/2019 Indonesian Aquaculture 2018, Jakarta, 28-30 November 2018 64

65. 0,084
0,095
0,098
0,138
0,070
0,080
0,090
0,100
0,110
0,120
0,130
0,140
0,150
0 50 100 150
Weight in Nursery
(gram)
Dose of Postbiotics (ppm)
10/28/2019 Indonesian Aquaculture 2018, Jakarta, 28-30 November 2018 65

66. 2,5
2,1
1,9
1,3
1,10
1,30
1,50
1,70
1,90
2,10
2,30
2,50
0 50 100 150
FCR in Nursery
Dose of Postbiotics (ppm)
10/28/2019 Indonesian Aquaculture 2018, Jakarta, 28-30 November 2018 66

67. 10/28/2019 Indonesian Aquaculture 2018, Jakarta, 28-30 November 2018 67
Partial Harvest 0.5-1 kg/m3 in Trial Tanks
1,1
5,1
4,4
7,0
1 2 3 4 5 6 7
800
600

68. Area 800 m2
Cycles 3 siklus/tahun
Tanks 10 @50 m3
DOC 78 days
Density 240 ind/m2
Biomassa 4 kg/m3
ADG 0,23 gram/day
SR >90%
FCR 1,1-1,2
Size 50-70 ind/kg
Shrimp Profile Production in Tanks

69. REMINDING THE LINE
Finding
stuck in
food
chain
Preparing
healthy
basic of the
pyramides
Helping
the
grazer
Controlling
the
environment

70. SHRIMPING
Let’s