Minazel Plus Aquaculture

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+ PATENT-CO.COM

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shrimps

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PATENT CO. invests in state-of-the-art equipment for the production of animal feed additives and premixtures, allowing us to achieve final products of a consistently high quality.

Mycotoxins... ... are recognized as a global threat in agricultural production, especially in livestock and aquaculture segment. There is an increasing trend to use more and more plant ingredients in aquaculture feeds. And with it the risk of mycotoxin contamination also increases.

growing crops

humans

harvesting

human food

storage

processing

fish and shrimps

distribution

feed

What is common to all mycotoxins is their negative effect and the direct impact on the economic losses in the production of aquaculture species. With the current negative trend of mycotoxin effects on the FCR of 5%, by 2020, an increase in feeding costs of $710 million is expected, in Marine Fish and salmonids feeding.

Did you know?

More than 25% of world crops are contaminated with mycotoxins!

25%

75%

Mycotoxin effects on aquatic species Biological effects of mycotoxins in aquatic species are thought to be directly linked to their concentration in the feed and to the animal's age and species. In aquaculture production, mycotoxin can cause serious negative effects on fish and shrimps. They can be caused by one mycotoxin, or by their synergistic effects which can cause more damage, even when present in low levels.

Aflatoxin B1

Ochratoxin A

Small cell carcinoma Oxidative damage 3 Pale gills 4 Reduction of growth, feed conversion rate, weight gain 5 Increasing mortality 6 Abnormalities in shrimp hepatopancreas.

1

2

2

1 Mutagenic and toxic effect Degeneration of kidneys and liver, followed by increased mortality 3 Poor feed conversion ratio

All mycotoxins Immunosuppression Increased mortality Poor productive performances 1

2

3

Reduced spawning frequency

Liver necrosis

Fumonisins

Pale kidneys and renal necrosis

Lesions in exocrine and endocrine pancreas Lesions in inter-renal tissue 1

2

Zearalenone 1

4

Affects reproductive parameters in different aquatic species 2 Reducing spawning frequency 3 Change of relative fecundity Acceleration of sexual maturation 5 Reduced growth in shrimp 6 Leaves residues in meat

Reduced growth, consumption and poor feed conversion

Trichothecenes 2

1 Decreased average weight gain Decreased production of bacterial cell wall breaking enzymes 3 Immunosuppression at low dosage 4 Decreased resistance to oxidative damage 5 Inhomogeneous growth and physiological disorders in shrimps

we have a solution!

Patented technology MINAZEL PLUS is a NEW COMPOUND created by patented technology, consisting of a: Mineral component

polar mycotoxin

polar mycotoxin Na1+ Ca2+

K1+

Mg2+

Na1+

Ca2+

K1+

Na1+ Ca2+

polar mycotoxin

polar mycotoxin

polar mycotoxin K1+

mineral surface

Easily replaced Cations (K, Ca, Na, Mg)

Mg2+ Ca2+ Na1+

Na1+ Ca2+

K1+

natural clinoptilolite surface (negative surface is caused by substitution of Si4+ with Al3+ or Fe3+)

The addition of organic cations serves to change the mineral surface. The result of this addition is not a simple mixture of mineral and organic phase, but a completely new compound, organo-mineral complex.

MINAZEL PLUS is the result of an ion-exchange reaction between inorganic cations on the mineral surface and organic cations.

Organic component

+ + +

+

+

less polar mycotoxins

less polar mycotoxins

Na1+ Ca2+

K1+

Mg2+

Na1+

less polar mycotoxins

less polar mycotoxins

polar mycotoxin

polar mycotoxin

less polar mycotoxins

Ca2+

K1+

polar mycotoxin Na1+ Ca2+

K1+

polar mycotoxin Mg2+ Ca2+ Na1+

polar mycotoxin Na1+ Ca2+

K1+

MINAZEL PLUS

Less Polar Mycotoxins Binding (Zea, Ochra, T-2...)

organic component

Polar Mycotoxin Binding (Afla, Ergot...)

mineral surface

Chains of Organic Cation (Activation)

New active centres, which are formed on the mineral surface, ensure efficient binding (over 90%) of not only POLAR MYCOTOXINS (Aflatoxins, Ergot Alkaloids, etc.) but also of LESS POLAR MYCOTOXINS (Zearalenone, Ochratoxin A, T-2 toxin, etc.).

MINAZEL PLUS is highly effective (IN VITRO trial results): 100%

99%

97%

96%

94%

86%

75%

83%

80%

50%

25%

in cit rin

s er go

ta

lka

lo

id

n to xi T2

n fu

oc

m

on

isi

hr at ox

in

B1

A

ne en o ra l ze a

afl at ox

in

B1

0%

adsorption %

AB1

AB1

90.2%

99.6%

100%

79.7%

86.6%

83.7%

5 min

30 min

60 min

5 min

30 min

60 min

contact time

ZON

OTA

78.5%

83%

93.2%

79.5%

86.3%

87.4%

5 min

30 min

60 min

5 min

30 min

60 min

MINAZEL PLUS - speed of adsorption (LC-MS/MS)

MINAZEL PLUS trial reports in aquatic species PRODUCTION PARAMETERS IN COMMON CARP Cyprinus carpio DURING DIETARY APPLICATION OF MINAZEL PLUS MYCOTOXIN ABSORBER Dušan Palić 1, Ksenija Aksentijević 2, Marko Vasiljević 3

Munich, Germany, 2016

1 Chair for Fish Diseases and Fisheries Biology, Faculty of Veterinary Medicine, Ludwig-Maximilians-University Munich, Germany 2 Department of Microbiology, Faculty of Veterinary Medicine, University of Belgrade, Serbia 3 PatentCo, Mišićevo, Serbia

The fish were reared in recirculation systems (400 L tanks) with automated 30% water exchange per day. Water quality parameters were monitored daily or weekly.

Two hundred fish were randomly assigned to treatment with MINAZEL PLUS and experimental (no additive) groups. The fish were sampled on regular basis to collect biometric parameters (Fig. 1.), perform clinical health and histopathological exam, and determine total and differential leukocyte counts.

Average body weight (g)

45

Effect of MINAZEL PLUS on Common Carp Body Weight

40 35

MINAZEL PLUS 2kg/MT

30

Control

25 20

Start

7

14

21

28

35

42

50

Trial duration (days)

Figure 1. Effects of MINAZEL PLUS additive (2kg/MT of MINAZEL PLUS) on body weight in common carp. *denotes significant differences at p<0.05.

Conclusion Fish in experimental group showed significant increase in body weight (Fig 1.) and decrease in feed conversion ratio. Clinical and pathological findings were not significantly different among treatment and control groups. We conclude that addition of 2kg/MT of MINAZEL PLUS to commercial fish diet significantly increased production parameters in common carp without observable side effects.

EFFECTS OF MINAZEL PLUS USED AS FEED ADDITIVE ON PRODUCTION PARAMETERS IN WHITELEG SHRIMP Litopenaeus vannamei Munich, Germany, 2016

Dušan Palić 1 , Eman Zahran El Naggar 2, Marko Vasiljević 3

1 Chair for Fish Diseases and Fisheries Biology, Faculty of Veterinary Medicine, Ludwig-Maximilians-University Munich, Germany 2 Department of Internal Medicine, Infectious and Fish Diseases, Faculty of Veterinary Medicine, Mansoura University, Egypt 3 PATENT CO, Misicevo, Serbia

Shrimp (200/group) were housed in two 1,000 liter tanks connected to a large commercial shrimp production recirculation system (Crustanova®). The shrimp were sampled on regular basis to collect biometric parameters (Fig. 2.), perfrom clinical health exam, histopathology exam and determine total and differential leukocyte counts.

Effect od MINAZEL PLUS on Shrimp Body Weight

average body weight (g)

24

19

MINAZEL PLUS 2kg/MT

14

Control

9

4

Start

20

45

55

trial duration (days)

Figure 2. Effects of MINAZEL PLUS additive (2 kg/MT of MINAZEL PLUS) on body weight in Pacific white legged shrimp;

Initial weight [g]

MINAZEL PLUS 4.14

22.16

19.66

18.10

0.40

0.37

3931.97

3619.53

1.57

1.70

MINAZEL PLUS CV% weight CV% length

Control CV% weight CV% length 13.78 36.87 14.71 44.79 8.33 20.64 10.75 30.63

26.27

9.81

23.54

9.27

07.12.2016

18.36

12.17

19.12.2016

20.85

7.73

25.10.2016 15.11.2016

4.06

23.80

Final weight [g] Weight gain [g] ADG (Average daily gain) [g] Mass weight gain [g] Food Conversion Ratio

Coefficient of Variation (%)

Control

Table 1. Pacific white-legged shrimp biometric parameters are presented.

Conclusion Group fed with addition of 2 kg/MT of MINAZEL PLUS has significantly better parameters as seen in the table.

ORGANICALLY MODIFIED CLINOPTILOLITE (MINAZEL PLUS®) FEED ADDITIVE IMPROVES HEALTH AND PERFORMANCE IN CULTURED NILE TILAPIA Nemanja Todorović 1, Marko Vasiljević 1, Eman Zahran El-Naggar 2 and Dušan Palić 3 Mansoura, Egypt, 2017 1 PATENT CO. Mišićevo, Serbia 2 Department of Internal Medicine, Infectious and Fish Diseases, Faculty of Veterinary Medicine, Mansoura University, Egypt 3 Chair for Fish Diseases and Fisheries Biology, Faculty of Veterinary Medicine, Ludwig-Maximilians-University Munich, Germany

Testing efficacy of MINAZEL PLUS protection against low level (16 µg/g) Aflatoxins (AFT) contamination in feed: 14 tilapia (45g average weight) per group were randomly assigned to control, MİNAZEL PLUS, AFT, AND AFT W/ MİNAZEL PLUS groups. Samples were collected for evaluation of hematological, immunological, pathological and enzymatic parameters.

Degranulation of Primary Granules

15

70

13

60

% MPO release

average body weight (g)

Effect of MINAZEL PLUS on Tilapia Body Weight

11 9 7

30 20

5

10

3

0

1

Start 7

14

21

28

33

40

47

61

75

90

9

Activity index

Effect of MINAZEL PLUS on Tilapia Body Length

Start

8 7

1.8 1.6 1.4 1.2 1.0 0.8 0.6 0.4 0.2 0.0

6

Start

5

90th day

35th day

90th day

Oxidative Burst Start 7

14

21

28

33

40

47

61

75

trial duration (days) MINAZEL PLUS 2kg/MT

Control

Tilapia growth is significantly increased in group fed with addition of 2kg/MT of MINAZEL PLUS. A) Weight gain B) Length in tilapia juveniles of 2 g starting weight.

90

Activity index

4

35th day

Neutrophil release

trial duration (days)

average body length (g)

50 40

1.8 1.6 1.4 1.2 1.0 0.8 0.6 0.4 0.2 0.0

Start

35th day

90th day

MINAZEL PLUS 2kg/MT Control

Neutrophil function in tilapia from MINAZEL PLUS group was significantly increased

Aflatoxin treated group of tilapia was diagnosed with normocytic and normochromic anemia, while such pathology was not observed in group with the addition of MINAZEL PLUS, indicating additive ability to rescue fish from negative effects of aflatoxin.

Conclusion We conclude that addition of 2kg/MT of MINAZEL PLUS to commercial fish diet significantly increased production parameters in Nile tilapia without observable side effects, and protected fish health during exposure to low level aflatoxin contamination.

PROTECTIVE EFFECT OF ORGANICALLY MODIFIED CLINOPTILOLITE (MINAZEL PLUS) IN FISH EXPOSED TO MYCOTOXINS AND CHALLENGED WITH INFECTIOUS DISEASE Nemanja Todorović 1, Marko Vasiljević 1, Dušan Palić 2

Mansoura, Egypt, 2017

1 PATENT CO. Mišićevo, Serbia 2 Chair for Fish Diseases and Fisheries Biology, Faculty of Veterinary Medicine, Ludwig-Maximilians-University Munich, Germany

TRIAL SETUP Treated group

Investigated groups

Fathead minnow (Pimephales promelas)

Duration

1. No treatment

14 days

2. MINAZEL PLUS only

Infectious disease challenge*** 14-28 days

3. AFTB1/OTA* 4. AFTB1/OTA/*MINAZEL PLUS**

* 50 µg/g Aflatoxin B1 and 8 µg/g Ochratoxin A ** MINAZEL PLUS used in dosage 5kg/MT *** Challenge with Aeromonas hydrophila (6 x 107 CFU/mL, 10 µL g-1 of fish body weight)

RESULTS ON MORTALITY, MORBIDITY AND RPS (RELATIVE PERCENT SURVIVAL) GROUPS T1

DAY

T2

T3

Healthy Fish

Live Fish

Healthy Fish

14

60

60

58

21

38

50

22

28

24

45

12

24

Morbidity

36

Live Fish

48

T4

Healthy Fish

Live Fish

58

59

59

30

45

55

33

50

25

27

Healthy Fish

T5 Live Fish

Healthy Fish

Live Fish

59

59

60

60

36

48

59

59

43

59

59

35

0

Mortality

15

36

10

17

1

RPS

75%

40%

83.3%

71.7%

98.3%

T1 - Control Diet with A. hydrophila Challenge T2 - Aflatoxin B1 50 µg/g + Ochratoxin A 8 µg/g T3 - MINAZEL PLUS 5kg/MT

T4 - Aflatoxin B1 50 µg/g + Ochratoxin A 8 µg/g with MINAZEL PLUS 5kg/MT

T5 - Control Diet w/o A.hydrophila Challenge

Conclusion Addition of MINAZEL PLUS significantly reduced morbidity and mortality of minnows fed with mixture of mycotoxins and challenged with A. Hydrophila!!!

ABOUT US PATENT CO. is a multinational company established in 1990 in the heart of Europe, in Serbia. Today we operate on 5 continents.

PATENT CO. strives to be an innovative and internationally-oriented company that is always one step ahead in introducing new technologies in animal nutrition.

RESEARCH AND DEVELOPMENT Every year, PATENT CO. invests significant resources in research and development programmes in laboratories, research centres and farms.

This programme facilitates the development of new products, with a view to finding optimal animal feed production solutions.

QUALITY AND SAFETY PATENT CO. invests in state-of-the-art equipment for the production of animal feed additives and premixtures, allowing us to achieve final products of a consistently high quality. This process ensures full traceability, from the reception of raw materials to delivery of the product to the customer.

Vlade Ćetkovića 1a 24211 Mišićevo, Serbia www.patent-co.com export@patent-co.com