Livestock Research for Rural Development 24 (6) 2012 Guide for preparation of papers LRRD Newsletter

Citation of this paper

Detoxifying effects of a commercial additive and Phyllanthus amarus extract in pigs fed fumonisins-contaminated feed

Nguyen Hieu Phuong, Brian Ogle*, Petterson H and Nguyen Quang Thieu

Faculty of Animal Science and Veterinary Medicine,
Nong Lam University, Ho Chi Minh City, Vietnam
phuongnguyen180984@yahoo.com
Department of Animal Nutrition and Management, Swedish University of Agricultural Sciences,
PO Box 7024, S-750 07, Uppsala, Sweden

Abstract

Reduction of the negative effects fumonisims in pigs of a Phyllanthus amarus extract and a commercial detoxifying additive product was evaluated with respect to growth performance, pathology and blood biochemistry. Forty eight crossbred (Landrace × Yorkshire × Duroc) weanling pigs were randomly assigned in a completely randomized design (CRD) to six diets containing: 1) low fumonisin B1 and no feed additive (LFNA); 2) low fumonisin B1 and a commercial detoxicant additive at 1g/kg of feed (LFCA); 3) low fumonisin B1 and Phyllanthus amarus extract at 10g/kg of feed (LFPE); 4) high fumonisin B1 and no feed additive (HFNA); 5) high fumonisin B1 and commercial detoxicant additive at 1g/kg of feed (HFCA); 6) high fumonisin B1 and Phyllanthus amarus extract at 10g/kg of feed (HFPE).
Fumonisin levels, detoxicants and their combination did not have any effect on final weight, average daily weight gain, average daily feed intake, and feed conversion ratio in pigs. Including 3980 µg fumonisins/kg diet decreased the total cholesterol significantly compared with the low fumonisin groups (2.19 mmol/L < 2.42 mmol/L) (P<0.05). The aspartate aminotransferase (AST) blood levels of pigs given the commercial additive were higher than in the no feed additive group (110 U/L > 83.1 U/L, P=0.067) and the Phyllanthus amarus group also had a high AST blood level (99.6 U/L). Moreover, fumonisins thickened the alveolar walls of the lungs, while the commercial and Phyllanthus amarus additives partly reduce the thickened alveolar wall lesions. Liver cells also had more severe fatty degeneration and necrosis in the fumonisin and no additive group than in the commercial and Phyllanthus amarus groups. However P. amarus extract made the liver tender.

Key words: Fumonisin B1, blood biochemistry, aspartate aminotransferase, AST, lung, liver


Introduction

Fumonisin presents one of the greatest potential mycotoxin risks to human and animal health, as a food and feed contaminant, along with aflatoxins, trichothecenes, zearalenone, ochratoxin A and ergot alkaloids (CAST  2003). There are four main types of fumonisin, B1, B2, A1 and A2, and among these fumonisin B1 is produced in the largest amount and has highest toxicity. These toxins are produced by Fusarium verticillioides (previously known as F. moniliforme) (Gelderblom  et al  1988). Fumonisins cause equine leukoencephalomalacia and pulmonary edema syndrome in pigs and horses (CAST  2003). Raviprakash  et al (1997) revealed that fumonisin B1 affected kidney weight and induced apoptosis in mouse liver. It is also reported that fumonisins could cause hepatocarcinoma in rats (Gelderblom  et al  2001). Fumonisins elevated some serum enzymes, such as AST, ALT, ALP and CREA (Creatinine) and caused an abnormality of liver histophathology (Zomborszky  et al  2002; Piva  et al  2005). Galvano  et al (2001) indicated that the prolonged exposure of FB1 at high concentration caused DNA damage of apoptotic type in human fibroblasts. In Vietnam, in earlier studie of Nguyen Quang Thieu at al. (2008) the occurrence of aflatoxins and zearalenone was widespread and in high concentrations. Especially, fumonisin incidence was about 70% in tested samples, and the highest fumonisin B1 level was around 10.8 ppm in the Southeastern and Highlands provinces (Phuong  et al  2010).
Since the occurrence of fumonisins is harmful to animal and human health, there are many methods to reduce their production or their negative impacts, such as using resistant corn varieties, controlling insects, heating, chemical detoxificants like Ca (OH)2 and ammonia, or degradation of fumonisins by ozone. Nevertheless, some of these methods are not very effective; some are successful but need high temperatures to hydrolyze fumonisins and other toxins (Maja Šegvić and Stjepan Pepeljnjak  2001). Using adsorbents to adsorb mycotoxins, especially aflatoxin, is also one of the methods that can effectively reduce the harmful effects. However, they do not really show any effects on fumonisins, and adsorbants such as activated carbon seem to worsen the toxic effect of fumonisin B1 (Piva  et al  2005), while cholestyramin also did not have a specific effect in adsorption of fumonisins in vivo (Solfrizzo  et al  2001). Some commercial products have adsorptive effects on fumonisins, but their costs are too high, particularly for small farmers. Besides, fumonisins after being absorbed through the intestine, will go to the liver first, and exert effects on it, and then on other organs.
According to Taylor (2003), indigenous peoples use plants from Phyllanthus sp., a small annual herb that is distributed throughout the tropical regions of the world, to cure many diseases including hepatitis and other liver diseases. In clinical research, this plant also demonstrated its anti-hepatotoxic function. A study by Huang  et al (2004) showed that the water extract of Phyllanthus urinaria, the same family and the same use as Phyllanthus amarus, could reduce the cell viability of HL-60 cells (human myeloid leukemia), which is additional evidence for its anticancer effect. This plant had the ability to reverse the negative impacts of aflatoxicosis of broiler chickens (Sundaresan  et al  2007). Phyllanthin, hypophyllanthin and niranthin are the main substances in Phyllanthus niruri that have a hepatoprotective effect (Kodakandla  et al  1985). Phyllanthus urinaria or Phyllanthus amarus is a weed that is easy to cultivate and is widespread. In Vietnam, Phyllanthus amarus grows widely in nature and has been used as a medicinal plant for a long time to treat hepatic disease, eye infection, snakebites and acnes (Do Tat Loi  2004). Today, many farmers in the poor Central provinces of Vietnam are earning money from growing this plant for use as human medicine (Hung Phien  2009). For these reasons, we can hope that adding Phyllanthus amarus or P. urinaria extract to the feed of pigs can reduce the effects of fumonisins on liver, and thus liver, kidney and lung functions will not be affected and the pigs will retain good production performance.


Materials and methods

Site description
The experiment was conducted in the experimental farm of the Animal Husbandry and Veterinary Medicine Faculty of Nong Lam University, Ho Chi Minh City, Vietnam. This area is in Southeastern Vietnam, and has a tropical monsoonal climate, with a rainy season from May to October and a dry season from November to April. The average temperature is 27.5°C with high humidity. The duration of this study was 37 days, from 22 December 2009 to 31 January 2010.
Pigs, treatments and experimental design
The experiment had a completely randomized design (CRD), with a 2 × 3 factorial arrangement and 4 replicates. Each replication had 2 pigs (one male castrate and one female) in one pen. In total 48 crossbred (Landrace × Yorkshire × Duroc) piglets (10.9 ± 1.21 kg initial body weight) were used in the experiment. The pigs had been vaccinated against mycoplasma and swine fever at one week of age and five weeks of age. After 4 days of acclimatization, the pigs were randomly assigned to the treatments and pens according to the design shown in Table 1 and Figure 1. The treatments were: 

Fumonisins factor:

Additive factor:

 

Table 1. Individual treatments

 

NA

CA

PE

HF

HFNA

HFCA

HFPE

LF

LFNA

LFCA

LFPE

                              

Figure 1. Experimental layout

Table 2. Composition of Phyllanthus amarus extract
Composition
Niranthin
Hypophyllanthin
Phyllanthin
Concentration (mg/g dry matter)
2.71
1.71
7.98

Fumonisin B1 production
Fumonisin B1 (FB1) was from two sources: fumonisin B1 produced by Fusarium proliferatum cultured on maize grains was purchased from the National I-Lan University Laboratory, Taiwan, and production by Fusarium proliferatum isolates that was assessed in cultures grown on autoclaved rice. In brief, 500 g of ground maize was placed in a jar, was moistened for 1 h in distilled water and then autoclaved at 1210C for 60 minutes.
The maize medium was inoculated with mycelia from seven-day-old cultures of Fusarium proliferatum grown on PDA (Potato Dextrose Agar) and incubated at 26oC for 15 days. Samples (three replicates) of each isolate were dried in a forced-air draft oven at 55oC for 48 h, crushed with a mortar and pestle, and extracted with 1 ml of CH3CN:H2O (1: 1, v ⁄ v) for 1h. The extracts were centrifuged at 1500g for 10 min, and the supernatants were cleaned on a Sep-Pak C18 cartridge column (Waters, Milford, MA, USA) with CH3CN:H2O (1 : 1, v ⁄ v) as the solvent. The level of FB1 was determined from the extracts using HPLC. The concentration of fumonisin B1 in the substrate was 882 ppm. A commercial source of pure fumonsin B1 was used with 5514 mg fumonisin B1 in 280.9g powder.
Detoxicant
A commercial detoxicant additive was used.  Phyllanthus amarus extract was purchased from Hong Dai Viet Company, Vietnam, and its components (phyllanthin, hypophyllanthin and niranthin) were analyzed at the Institute of Chemical Technology, Vietnam.

Diets

The diet was formulated to meet or exceed the nutrient requirement for pigs (NRC  1998) (Table 3) and was then mixed with fumonisin B1 to produce the diet that contained 10 mg FB1 per kg feed. Finally, detoxicant additives were mixed with feed according to treatments. Feed was analyzed for gross composition, including crude protein, ether extract, crude fibre, ash, Ca and P according to AOAC (1990).

Table 3. Ingredient composition of the basal diet, as fed

Ingredients

Amount (%)

Extruded maize

48.4

Extruded soybean

20.6

Rice bran

8.01

Soybean meal

7.01

Fish meal 60%

7.01

Lactose

5.01

Dicalciumphosphate

1.03

Fat powder

0.54

Whey powder

0.42

Lactic acid

0.40

Threonine

0.39

Vitamin and mineral premix

0.30

Lysine HCl

0.25

Choline 60%

0.20

Methionine

0.16

Salt

0.15

Colistin 10%

0.10

Total

100

Feed and drinking water were offered ad libitum and hygienic conditions maintained throughout the experiment. Fresh feed was provided at 08:00 h and 15:30 h, when remaining feed was removed and weighed.

Data collection
The animals were weighed at the beginning and the end of the trial to calculate average daily weight gain (ADG) and then feed conversion ratio (FCR) was calculated based on feed consumption. Blood samples of all pigs were collected at the end of the experiment and sent to MEDIC Medical Center, Ho Chi Minh City for analysis of bilirubin T, bilirubin D, bilirubin I, albumin, total cholesterol, glucose, AST (Aspartate Aminotransferase or GOT: Glutamic Oxaloacetic Transaminase), ALT (GPT: Glutamine Pyruvic Transaminase), GGT (Gamma Glutamyl Transpeptidase) and ALP (Alkaline Phosphatase).
At the end of the 37-day experiment, two pigs per treatment were slaughtered and necropsies performed. The lungs, liver and kidneys were sectioned to observe lesions and biopsies carried out to examine the histopathology.
Chemical analysis
The formulated feed was sampled and the composition of the basal diet was determined. The feed also was checked for fumonisins and aflatoxins concentrations. The concentrations of total aflatoxins in the feed were 6.3 and 13.5 µg/kg for LF and HF groups, respectively. Although the expected fumonisin B1 level in the diet was 10000 µg/kg, the analyzed fumonisins level was 3980 µg/kg (fumonisin B1: 2839 µg/kg; fumonisin B2: 1141 µg/kg) in the HF group. Moreover, the feed of LF group contained 1132 µg fumonisins/kg of feed (fumonisin B1: 693 µg/kg; fumonisin B2: 439 µg/kg).
Serum analysis was done by MEDIC Medical Center, Ho Chi Minh City. Histopathology examination was carried out by the Veterinary Hospital, Nong Lam University, Ho Chi Minh City.
Statistical analysis
The data were analyzed by analysis of variance (ANOVA) using the CRD procedure of Minitab software (Minitab 2000). Sources of variance were treatments and error.


Results

The chemical composition of the basal diet is shown in Table 4.
Table 4. Chemical composition of the basal diet, analysed values (% of DM, except for DM which is on air-dry basis))
 
Dry matter
Crude protein
Ether extract
Crude fibre
Ash
Ca
P
 

89.7

21.4

6.37

3.56

5.94

1.07

0.73

In order to ensure that the data gave exact results, the performance data of two pens from two treatments (LFCA and LFNA) in the experiment were not used in data analysis because these pigs were infected by Actinobacillus pleuropneumonia (APP) and depressed for a long period. Furthermore, the blood profiles of pigs from five pens from four treatments (two from LFPE, three from LFCA, HFNA, HFCA) were also not used in data analysis owing to identification mistakes in the laboratory. 

High fumonisins and adding detoxicants in the diets did not affect the growth performance (Table 5). Final weight, average daily weight gain, average daily feed intake, and feed conversion ratio were not significantly different among treatments (P>0.05) (Table 6).


Table 5. Effects of fumonisin level and commercial additive or Phyllanthus amarus extract on growth performance of weaned pigs
 
 Fumonisin
 Detoxicant 
 
Low
High
SEM
Prob.
NA*
CA*
PE*
SEM
Prob.
LW**, kg
 
 
 
 
 
 
 
 
 
Initial
11.1
10.8
0.223
0.374
10.9
10.7
11.3
0.288
0.456
Final
25.3
25.7
0.775
0.737
26.0
24.9
25.6
0.950
0.697
ADG**, kg
0.384
0.402
0.018
0.482
0.409
0.382
0.388
0.022
0.656
ADFI**, kg
0.735
0.741
0.028
0.883
0.744
0.735
0.735
0.035
0.979
FCR**
1.93
1.85
0.038
0.193
1.83
1.93
1.91
0.047
0.276
*NA: No detoxicant additive; CA: Commercial detoxicant additive; PE: Phyllanthus amarus extract
**LW: live weight; ADG: average daily weight gain; ADFI: average daily feed intake; FCR: feed conversion ratio

  

Table 6. Combination effects on piglet performance of commercial detoxifying additive or Phylanthus amarus extract with two levels of fumonisin in the diets

 

 

Low fumonisins

 

High fumonisins

 

 

 

NA*

CA*

PE*

NA*

CA*

PE*

SEM

Prob.

LW**, kg

 

 

 

 

 

 

 

 

Initial

11.1

11.2

11.0

10.6

10.3

11.5

0.407

0.282

Final

26.4

25.4

24.1

25.6

24.4

27.1

1.34

0.426

ADG**, kg

0.413

0.384

0.354

0.405

0.380

0.421

0.031

0.290

ADFI**, kg

0.788

0.733

0.683

0.699

0.736

0.788

0.049

0.164

FCR**

1.92

1.92

1.94

1.74

1.94

1.88

0.066

0.308

*NA: No detoxicant additive; CA: Commercial detoxicant additive; PE: Phyllanthus amarus extract

 Table 7 shows the differences in blood bilirubin T, D, I, albumin and total cholesterol among pigs given diets with different toxin levels and detoxicants. High fumonisins in the diet decreased the total cholesterol significantly (2.19 mmol/L) compared with the low fumonisin groups (2.42 mmol/L) (P<0.05). The differences in these parameters among the no feed additive, commercial additive and Phyllanthus amarus extract treatments were not significant (P>0.05). Moreover, the combination of detoxicant type and fumonisin level did not significantly affect bilirubin T, D, I, albumin and total cholesterol levels in pig blood (P>0.05) (Table 8). However, blood albumin in the high fumonisin - no feed additive group tended to be lower than in the other treatments (3.00 g/100mL).  

Table 7. Effects of fumonisin level and commercial additive or Phyllanthus amarus extract on serum blood parameters of piglets

 

Fumonisins

Detoxicants

 

Low

High

SEM

Prob

NA*

CA*

PE*

SEM

Prob

Bilirubin T, mg/100mL

0.381

0.430

0.032

0.298

0.466

0.380

0.370

0.039

0.179

Bilirubin D, mg/100mL

0.198

0.219

0.015

0.349

0.230

0.177

0.219

0.018

0.124

Bilirubin I, mg/100mL

0.183

0.197

0.025

0.685

0.216

0.203

0.150

0.032

0.309

Albumin, g/100mL

3.29

3.23

0.081

0.631

3.17

3.27

3.33

0.100

0.518

Tot Chol**, mmol/L

2.42

2.19

0.049

0.007

2.24

2.34

2.33

0.060

0.407

*NA: No detoxicant additive; CA: Commercial detoxicant additive; PE: Phyllanthus amarus extract

**Tot Chol: Total cholesterol

Table 8. Combination effects of commercial additive and Phyllanthus amarus extract on serum blood parameters of piglets fed fumonisin contaminated diets

 

Low fumonisins

 

 

High fumonisins

 

NA*

CA*

PE*

 

NA*

CA*

PE*

SEM

Prob.

Bilirubin T, mg/100mL

0.418

0.335

0.390

 

0.515

0.425

0.349

0.055

0.412

Bilirubin D, mg/100mL

0.218

0.160

0.218

 

0.242

0.193

0.221

0.026

0.852

Bilirubin I, mg/100mL

0.200

0.175

0.173

 

0.232

0.232

0.128

0.042

0.506

Albumin, g/100mL

3.34

3.14

3.38

 

3.00

3.40

3.29

0.140

0.129

Tot Chol**, mmol/L

2.40

2.45

2.41

 

2.08

2.23

2.26

0.084

0.602

*NA: No detoxicant additive; CA: Commercial detoxicant additive; PE: Phyllanthus amarus extract

**Tot Chol: Total cholesterol

 

Table 9. Effects of fumonisin level and commercial additive or Phyllanthus amarus extract on serum enzyme activities of piglets

 

 

Fumonisins

 

 

Detoxicants

 

 

 

 

Low

High

SEM

Prob

NA*

CA*

PE*

SEM

Prob

GGT**,U/L

66.1

60.1

7.23

0.569

56.5

67.8

65.0

8.85

0.631

ALP**,U/L

322

291

26.5

0.118

326

318

275

32.5

0.888

AST**,U/L

99.2

94.1

6.56

0.593

81.9

110

97.7

8.03

0.067

ALT**,U/L

65.6

63.6

3.96

0.733

58.3

68.7

66.7

4.85

0.285

*NA: No detoxicant additive; CA: Commercial detoxicant additive; PE: Phyllanthus amarus extract

**GGT: Gamma Glutamyl Transpeptidase; ALP: Alkaline Phosphatase; AST: Aspartate Aminotransferase (GOT: Glutamic Oxaloacetic Transaminase); ALT: Alanine Aminotransferase (GPT: Glutamine Pyruvic Transaminase)

Serum enzymes levels are presented in Table 9 and Table 10. Although there was no significant difference in blood GGT, ALP, AST and ALT levels between toxin levels and detoxicants and their combination, some trends were noted among treatment groups. For example, AST blood levels of pigs given the commercial additive were higher than in the no feed additive group (110 U/L > 83.1 U/L, P=0.067) and the Phyllanthus amarus group also had a high AST blood level (99.6 U/L). Furthermore, pigs given the high fumonisins only diet (HFNA) tended to have lower blood levels of GGT, AST and ALT.

Table 10. Combination effects of commercial additive and Phyllanthus amarus extract on serum enzyme activities in piglets fed fumonisin contaminated diets

 

 

Low fumonisin

 

High fumonisin

 

 

 

NA*

CA*

PE*

NA*

CA*

PE*

SEM

Prob.

GGT**,U/L

64.3

65.3

68.7

48.7

70.3

61.4

12.4

0.703

ALP**,U/L

373

288

304

279

349

246

45.7

0.238

AST**,U/L

90.1

115.4

92.1

73.7

105.1

103.4

11.3

0.476

ALT**,U/L

62.8

65.4

68.5

53.8

72.0

65.0

6.82

0.514

*NA: No detoxicant additive; CA: Commercial detoxicant additive; PE: Phyllanthus amarus extract

**GGT: Gamma Glutamyl Transpeptidase; ALP: Alkaline Phosphatase; AST: Aspartate Aminotransferase (GOT: Glutamic Oxaloacetic Transaminase); ALT: Alanine Aminotransferase (GPT: Glutamine Pyruvic Transaminase)

The results of the microscopic and macroscopic pathology of the experimental pigs are summarized in Table 11. The lesions found indicated mycoplasma infection in pigs, and there were some distinct points in the lungs and liver of pigs consuming fumonisin. Microscopic examination showed that fumonisin thickened the alveolar walls of the lungs. However, the groups that were treated with commercial and Phyllanthus amarus additives had less thickened alveolar walls than the no additive group. Moreover, in the pigs given fumonisins fatty degeneration and necrosis of the liver cells was observed and more severe degeneration of the hepatocytes was noted, especially in the groups given diets without additives. In addition, macroscopic observation seemed to show tenderness and swelling of the liver in the Phyllanthus amarus groups. 

Table 11. Pathology in organs of pigs fed high and low fumonisin contaminated feed with or without detoxicants

Treatments

Organs

Lung

Liver

Kidney

LFNA 

Macroscopically: interstitial pneumonia

Microscopically:  mild Mycoplasma

Macroscopically: signs of cirrhosis

Microscopically: mild hyperemia, but normal

Macroscopically: pale

Microscopically: mild hyperemia, mild hemorrhage at the renal cortex

 

LFCA

Macroscopically: signs of Actinobacillus pleuropneumoniae  (APP)

Microscopically: mild  mycoplasma

 

Macroscopically: signs of cirrhosis

Microscopically: normal

 

Microscopically: normal

 

LFPE

Macroscopically:  Pleuritis, pale, pulmonary edema

Microscopically: Mycoplasma (+)

Macroscopically: tenderness of liver, pale

Microscopically: normal

 

Microscopically: normal

 

HFNA

Macroscopically: focal inflammation on the right lung

Microscopically: thickening of alveolar walls, hemorrhage, Mycoplasma (+) , red hepatization of many lobules, pneumonitis

Microscopically: fatty degeneration of hepatocytes in many lobules

Macroscopically: degeneration

Microscopically: inflammation, hemorrhage in renal cortex and renal medulla

HFCA

Macroscopically: pulmonary edema, moderate interstitial pneumonia, hydrothorax, inflammation of apical lobe

Microscopically: thickening of alveolar walls but mild and no pneumonitis, Mycoplasma (+)

Macroscopically: mild swollen, pale

Microscopically: normal, hyperemia in some lobules, mild fatty degeneration

Macroscopically: swollen, pale

Microscopically: normal

HFPE

Macroscopically: Pulmonary necrosis, atelectasis - carnification, Mycoplasma

Microscopically: thickening of alveolar walls but mild- serofibrinous pneumonia, Mycoplasma (+), many leukocytes

Macroscopically: tenderness of liver, necrosis

Microscopically: normal

 

Microscopically: normal

 


Discussion

Growth performance
The results in this study are in contrast with the results from Piva  et al (2005), which showed a significant decrease of ADG and a significant increase FCR of piglets fed diets that contained 30 ppm fumonisins given for 42 days. However, there was no difference in the ADFI of these pigs among treatments. Rotter  et al (1996) found out that average daily weight gain of male pigs fed a 10 ppm pure fumonisin B1 diet through 8 weeks decreased by 11% compared to a 0 ppm diet. Furthermore in the first 4 weeks of the experiment, a general increase of feed consumption was observed. Nevertheless, in agreement with growth performance results in the present study, Osweiler  et al (1992), and Zomborszky  et al (2002) did not find any difference in the feed intake, body weight gain, and feed conversion ratio of pigs fed a 10 ppm fumonisins diet and a 17 ppm fumonisin B1 diet. Possibly, the concentration of fumonisin B1 was not high enough and and time on experiment sufficiently long to cause any changes in the growth performance of pigs. In addition, the fumonisin B1 in this study was a mixture of pure and cultured FB1, and therefore the results were not as clear as those found by Rotter  et al (1996).
Blood parameters
While high fumonisins in the diet lowered total serum cholesterol in the present experiment, Rotter  et al (1996) demonstrated an increase in cholesterol serum after 2 weeks. When fumonisins were fed at 30 ppm, cholesterol concentration was significantly higher than in control pigs (Piva  et al  2005). On the other hand, the cholesterol and serum enzyme levels were within the normal ranges in the study of Zomborszky  et al (2002), and only some pigs had histopathological changes (fumonisins dose 1 and 5 ppm) and showed an elevation outside the normal ranges of AST, ALT and ALP in serum (ALP >500 U/ l, AST >100 U/ l and/or ALT >70 U/ l). Furthermore, a dose of 17 ppm fumonisin B1 in the diet did not show clearly any increase in AST, GGT and total bilirubin of pigs (Osweiler  et al  1992). High fumonisins concentration in the diet in the present study decreased the total cholesterol concentration (2.19 mmol/L) to values under the normal range of total cholesterol of from 2.36 to 3.72 mmol/L (Tumbleson and Kalish  1971). It was reported that the apparent digestibility of the ether extract (EE) of 10 mg FB1/kg feed was significantly reduced during the weanling phase, and then increased to the level in the control group after this phase (Gbore and Egbunike  2007). Because fumonisins are mentioned in the disruption of lipid metabolism action in animals, this suggests that 3980 µg fumonisins/kg feed decreased the cholesterol concentration in pig serum, and that this may be related to the lower digestibility of the ether extract over the 37 days of experiment. This mixture level and the length of experiment may not have been enough to increase the total cholesterol serum, as reported in some studies. However, the commercial additive and Phyllanthus amarus extract could have increased the cholesterol level (Table 7 and 8).
In terms of enzyme activity, the commercial additive and Phyllanthus amarus extract elevated the concentration of AST to higher levels than the safe limit (AST >100 U/L) (Zomborszky  et al  2002), especially, the commercial additive.
Histopathology
The observed microscopic pathology of lung is in the present study consistent with Osweiler  et al (1992) and Zomborszky  et al (2002), who found thickening of alveolar walls. Moreover, the liver histopathology is also in agreement with Gelderblom  et al (1988) concerning the degeneration of hepatocytes. It can be concluded that the commercial additive and Phyllanthus amarus extract had a mild effect in reducing the negative impacts of fumonisins. However, possibly because of the high dose, P. amarus extract caused tenderness of liver and AST elevation. Furthemore, the commercial additive also showed an AST level increase over the limit which can cause liver damage.


Conclusions


Acknowledgements

We wish to thank SIDA-SAREC for funding this research through the regional MEKARN project.


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Received 4 April 2012; Accepted 26 May 2012; Published 1 June 2012

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