IN VIVO TRAILS ON PIGS

IN VIVO TRAILS ON PIGS

HAEMATOLOGICAL AND BIOCHEMICAL PARAMETERS OF WEANED PIGLETS FED ON FODDER MIXTURE CONTAMINATED BY ZEARALENONE WITH ADDITION OF MINAZEL PLUS Acta Veterinaria (Belgrade), Vol. 56, No. 2-3, 121-136, 2006. ŠPERANDA MARCELA*, LIKER B**, ŠPERANDA T*, ŠERIĆ V***, ANTUNOVIĆ Z*, GRABAREVI]Ć****, SENČIĆ Đ*, GRGURIĆ D***** and STEINER Z* *Faculty of Agriculture, Osijek, Croatia; **Faculty of Agriculture, Zagreb, Croatia; ***Clinical Hospital Osijek, Croatia; ****Faculty of Veterinary Medicine, Zagreb, Croatia; *****Centar for Livestock Improvement, Osijek

Influence of Zearalenone on general condition of organs and the organism Influence of Zearalenone on metabolic status of piglets Reduction of harmful effects of Zearalenone through application of Minazel Plus, based on biochemical and haematological values and histological results

The research was conducted on 4 groups of 5 female piglets aged from 40 to 54 days. First control group (C1) received a concentrate without added Zearalenone or Minazel Plus. Second control group (C2) received a concentrate without added Zearalenone and with 0.2% of Minazel Plus. First experimental group (E1) received concentrate with 3 mg/kg of Zearalenone and 0.2% of Minazel Plus. Second experimental group (E2) received concentrate with 3 mg/kg of Zearalenone and without Minazel Plus. C1 C2 E1 E2 3 mg/kg of Zearalenone - - + + 0.2% of Minazel Plus - + + -

During the study following parameters were analyzed: Body weight of piglets Haematological values and biochemical indicators (blood samples were taken on the 8th and 14th day) in order to determine: The levels of metabolites (glucose, urea, creatinine, cholesterol, bilirubin, total proteins, albumins, triglycerides) Enzyme activities (aspartate amino transferase-ast, alanine aminotransferase-alt, alkaline phosphatase- ALP, gamaglutamyltransferase-ggt, cholinesterase-che, creatine kinase-ck, lactate dehydrogenase-ldh and amylase) Mineral levels (iron-fe) Blood cell count (number of erythrocytes, leukocytes, trombocytes) Levels of haemoglobin and haematocrit Histological changes

GROUP START FINAL Average Value SD Average Value SD C1 13.11 1.63 16.49 1.46 C2 13.68 1.55 16.93 1.76 E1 12.86 2.18 16.64 1.52 E2 12.71 2.05 15.70 2.60 (C1) without Zearalenone & without Minazel Plus (C2) without Zearalenone & with Minazel Plus (E1) with Zearalenone & with Minazel Plus (E2) with Zearalenone & without Minazel Plus

Parameter Glukose, mmol.l-1 Urea,mmol.L-1 Creatinin, mmol. L-1 Bilirubine-total, mmol L-1 Total proteins, g L-1 Albumine, g.l-1 Holesterole, mmol.l-1 Triglicerides, mmol.l-1 Fe,mmol.L-1 Days GROUPS C 1 (a x ± SEM) C2 (b x ± SEM) E1 (c x ± SEM) E2 (d x ± SEM) 8 5.33±0.37*E1,E2 4.30±0.40 3.92±0.45*C1 3.72±0.40*C1 14 5.05±0.37*C2,E2 3.58±0.40*C1 4.17±0.45 3.66±0.40*C1 8 4.53±0.40*E1 5.38±0.43 6.17±0.49*C1 5.22±0.43 14 5.20±0.40*C2 3.60±0.43*C1 4.80±0.56 4.35±0.49 8 62.33±7.77 69.80±8.52 63.75±9.52 70.40±8.52 14 61.16±7.77 45.80±8.52 50.50±9.52 62.50±9.52 8 5.66±17.59 4.40±19.27 4.75±21.54 4.80±19.27 14 5.33±17.59 3.60±19.27 3.75±21.54 5.50±21.54 8 48.25±2.52**C2,E1,E2 62.20±2.76**C1 58.25±3.09*E2,**C1 61.50±2.76*E1, **C1 14 48.51±2.52*C2,**E2 58.38±2.76*C1 54.92±3.09**E2 64.42±2.76**C1,E1 8 34.15±2.77 28.84±3.03 30.00±3.98 27.80±3.03 14 33.45±2.77 27.88±3.03 29.27±3.98 29.86±3.03 8 1.81±0.59 1.93±0.65 1.53±0.72 1.99±0.65 14 1.77±0.59 2.55±0.65 2.71±0.89 2.96±0.65 8 1.18±0.07**C2,E1,E2 0.46±0.08**C1 0.52±0.09**C1 0.33±0.08**C1 14 0.95±0.07**C2,E1,E2 0.47±0.10**C1 0.57±0.10**C1 0.33±0.09**C1 8 23.80±1.36**C2,E1,E2 12.08±1.49**C1,E2 13.10±1.66**C1,E2 4.80±1.49**C1,C2E1 ANEMIA 14 21.98±1.36**C2,E1,E2 8.33±1.92**C1 7.66±1.92**C1 4.67±1.66**C1

Enzyme U/L-1 AST ALT ALP GGT CHE AMY CK LDH Days GROUPS C 1 (a x ± SEM) C2 (b x ± SEM) E1 (c x ± SEM) E2 (d x ± SEM) 8 24.66±5.99*E2,**E1 37.20±6.57**E1 77.50±7.34**C1,C2,E2 52.20±6.57*E1,C1 14 38.00±5.99**E2 37.00±8.48**E2 47.00±8.48**E2 83.25±7.34**C1,C2,E1 LIVER FAILURE 8 30.66±2.68 36.80±2.94 40.75±3.29 32.00±2.94 14 33.83±2.68 36.75±3.29 30.33±3.79 36.75±3.29 8 182.33±16.86*E1 153.20±18.47 109.75±20.65*C1 134.80±18.47 14 157.33±16.86*C2,E2 78.50±20.65*C1 103.33±23.84 71.75±20.65*C1 8 20.66±5.05*E2 45.20±5.53 39.75±6.18 48.60±5.53*C1 14 36.50±5.05 47.33±7.14 38.3±7.14 51.25±6.18 8 682.66±39.93 638.20±43.74 568.00±48.90 619.00±43.74 14 706.33±39.93*E1,E2 586.00±56.47 526.66±56.47*C1 520.75±48.90*C1 8 1640.33±199.09 1916.40±218.09 1392.75±243.83 1261.60±218.09 14 1572.16±199.09 1484.50±243.83 1158.00±281.55 870.75±243.83 8 175.83±293.83*E1 233.60±321.88*E1 1440.00±359.87*C2,C1 615.20±321.88 14 357.83±293.83**E2 MUSCULE PROBLEMS 298.60±321.88**E2 488.75±359.87**E2 2162.40±321.88**C2, C1,E1 8 378.00±194.64 785.00±212.22 752.25±238.39 823.80±213.22 14 353.50±194.64*E2 758.20±213.22 891.50±238.39*C1 1078.20±213.22*C1 AST-aspartate amino transferase; ALT- alanine aminotransferase; ALP-alkaline phosphatase; GGT- gamaglutamyltransferase; CHE- cholinesterase; CK- creatine kinase; LDH- lactate dehydrogenase; AMY- amilase

Histopatological findings Groups C1 C2 E1 E2 Hepatitis interstitialis 0/0 0/0 1/5 4/5 LIVER INFLAMATION Depletio lymphocitaria lienis et lymphonodulli 0/0 0/0 2/5 5/5 Many secondary oocytes 0/0 0/0 0/0 5/5 Hyperplasia glandularis uteri 0/0 0/0 1/5 5/5 REPRODUCTION TRACT PROBLEMS Primary follicles on ovaries 2/5 2/5 3/5 5/5 (C1) without Zearalenone & without Minazel Plus (C2) without Zearalenone & with Minazel Plus (E1) with Zearalenone & with Minazel Plus (E2) with Zearalenone & without Minazel Plus

Piglets fed with Zearalenone contaminated feed (E2) have shown: Sex organ pathological changes (ovaries and uterus) Lymphocyte depletion in lymph glands Interstitial inflammation of liver Muscle inflammation Significantly lower Iron level in blood serum Piglets fed with Zearalenone contaminated feed with addition of Minazel Plus (E1) have shown: Negligible changes to sex organs No pathological changes of liver and muscles No influence on Iron level in blood serum Minazel Plus has successfully prevented negative effects of Zearalenone!

RESULTS OF MINAZEL PLUS APPLICATION IN SOWS, PIGLETS AND FATTENING PIGS FARM IN RUMENKA, SERBIA AVAKUMOVIĆ ĐORĐE PhD, member of Veterinary Academy of Science Prof. Dr VIDOVIĆ VITOMIR PhD, Faculty of Agriculture, University of Novi Sad

To investigate efficiency of Minazel Plus in prevention of mycotoxicosis caused by: Aflatoxin B1 Ochratoxin A Zearalenone

Trial was done with 2 groups of 11 gestating sows After farrowing trial was continued with born piglets, and fattening pigs respectively. Feed was contaminated with: 3,84 mg/kg of Zearalenone 0,032 mg/kg of Aflatoxin B1 0,38 mg/kg of Ochratoxine A In feed for trial groups 0.2% of Minazel Plus was added.

During the sow trial following parameters were measured: Number of litters with Vulvovaginitis Number of weak piglets Number of litter with diarrhea occurrence Mortality rate Average daily dain Number of weaned piglets per litter During the piglet and pig trial following parameters were measured: Mortality rate Average daily gain Feed conversion ratio

Parameter Trial Group Control Group Difference % Liveborn piglets per litter 11,54 10,81 +6,75% Average live weight of piglets, kg 1,35 1,2 +12,5 Litters with vulvovaginitis 0 4 Weak piglets 3 4-25% Mortality rate 3,15 3,36-6,25% Average weaning weight, kg 8,2 7,9 +3,8 Weaned piglets per litter 11,18 10,45 +6,98

Parameter Trial Group Control Group Difference % Average starting weight of piglets, kg 8,2 7,9 +3,8 Average ending weight of piglets, kg 26,8 24,8 +8,0 Average daily gain, kg 0,388 0,352 +10,22 Feed conversion ratio 2,05 2,39-14,23

Parameter Trial Group Control Group Difference % Average starting weight of pigs, kg 26,8 24,8 +8,0 Average ending weight of pigs, kg 106,3 102,0 +4,2 Average daily gain, kg 0,800 0,770 +3,9 Feed conversion ratio 2,78 2,86-6,3

From the trials it can be concluded that the application Minazel Plus at a concentration of 0.2% in feed: Effectively prevented occurrence of mycotoxicosis Had positive effect on the health of pigs in all categories Was beneficial in reducing the feed conversion ratio Had better growth Had better economic results

RESULTS OF MINAZEL PLUS APPLICATION IN SOWS FARM IN PANČEVO, SERBIA AVAKUMOVIĆ ĐORĐE PhD, member of Veterinary Academy of Science Prof. Dr VIDOVIĆ VITOMIR PhD, Faculty of Agriculture, University of Novi Sad

To investigate efficiency of Minazel Plus in prevention of vulvovaginitis and diarrhea occurrence in piglets, through treatment of sows.

Trial was performed on the farm with vulvovaginitis and diarrhea occurrence. Trial was performed with 98 sows divided in 6 groups, 3 trial and 3 control group. Sow feed was contaminated with 1.7 mg/kg of Zearalenone on average. In feed for trial groups 0.2% of Minazel Plus was added.

During the trial following parameters were measured: Number of litters with Vulvovaginitis Number of litter with diarrhea occurrence Mortality rate Average daily dain Number of weaned piglets per litter

Description Tral I Trial II Trial III Total Difference % T-I C-I T-II C-II T-III C-III T C Number of farrowing 18 17 15 18 15 15 48 50 Litters with vulvovaginitis 3 17 0 18 0 13 3 48-93,75% Litters with diarrhoea 1 12 2 15 1 8 4 35-88,6% Number of dead piglets 19 55 18 42 22 36 59 133 Mortality % 10.6 30.7 11.2 2.8 14.0 22.5 11.8 25.4-53,6% T Trial group C Control group

From the trials it can be concluded that the application Minazel Plus at a concentration of 0.2% in lactating sow feed considerably reduced: Number of litters with vulvovaginitis Occurrence diarrhea in suckling piglets Mortality rate

Effect of Minazel Plus in diet on scatole levels in back fat of pigs Dr Smiljana Raičević* PhD Thesis * University of Belgrade, Faculty of Veterinary Medicine Belgrade 2000.

Although fattening of young boars is more economical than fattening of castrates, problem arrives when it comes to sexual odor of boars which is well know defect of pork. Organoleptic characteristics, especially smell and taste, the consumers are the most important and most transparent indicators of quality meat. Skatole is one of the main carrier of sexual odor of pigs, is product of bacteria from digestive tract of pigs( ceacum and colon). After adoption from intestines, via the blood comes to all the tissues and due to its lipophilic nature shall be deposited in the adipose tissue.

to investigate the effect of Minazel Plus in diet on the skatole level in back fat of pigs

1. to examine the content of skatole in adipose tissue of pigs slaughtered in industrial slaughterhouses 2. examine the influence of Minazel Plus added to feed (0.5%) on skatole in adipose tissue of young boars and gilts 3. To determine the deposition of skatole in the content of the digestive tract (colon) of young boars fed with and without the addition of 0.5% of Minazel Plus

Number of pigs: 213 Age: 5-12 months Body weight: 53-110kg Control group feed: standard pelleted diet with 16% protein Experimental group feed: standard pelleted diet with 16% protein + 0,5% Minazel Plus

Adipose tissue was sampled from the lumbar part of refrigerated carcasses of all animals in the experiment The determination of skatole: 1. in adipose tissue of pigs- colorimetric method by protocol of Mortensen & Sorensen, 1984. 2. in intestinal content- High-performance liquid chromatography (HPLC)

Skatole conc. μ/g Skatole conc. μ/g Average concentration of skatole in adipose tissue of young boars Average concentration of skatole in intestinal content of young boars

Addition of 0,5% Minazel Plus in feed, decreased average concentration of skatole in adipose tissue of young boars by 41% Average concentration of skatole in intestinal content increases for 52,6% during fed with feed with added 0,5% of Minazel Plus. Reduction of skatole in adipose tissue of young boars during use of 0,5% Minazel Plus in feed is due the ability of Minazel Plus to binds a polar compounds such as skatole.