Growth performance, haematology and serum biochemistry of
Growth performance, haematology and serum biochemistry of
female rabbits (
female rabbits (
Oryctolagus cuniculus
Oryctolagus cuniculus
) fed dietary fumonisin
) fed dietary fumonisin
Francis A. GboreFrancis A. Gbore1,21,2*, and Olatunbosun Akele*, and Olatunbosun Akele33
1
1School of Biological Sciences, Universiti Sains Malaysia, Pulau Pinang, MalaysiaSchool of Biological Sciences, Universiti Sains Malaysia, Pulau Pinang, Malaysia 2
2Department of Environmental Biology and Fisheries, Adekunle Ajasin University, Akungba-Akoko, NigeriaDepartment of Environmental Biology and Fisheries, Adekunle Ajasin University, Akungba-Akoko, Nigeria 3
3Microbiology Laboratory, Department of Microbiology, Adekunle Ajasin University, Akungba-Akoko, NigeriaMicrobiology Laboratory, Department of Microbiology, Adekunle Ajasin University, Akungba-Akoko, Nigeria
GBORE, F. A., O. AKELE:
GBORE, F. A., O. AKELE: Growth performance, haematology and serum Growth performance, haematology and serum biochemistry of female rabbits (
biochemistry of female rabbits (Oryctolagus cuniculusOryctolagus cuniculus) fed dietary fumonisin. Vet. ) fed dietary fumonisin. Vet.
arhiv 80, 431-443, 2010. arhiv 80, 431-443, 2010.
ABSTRACT ABSTRACT
To account for the potential effects of dietary mycotoxin on growth performance, haematological and To account for the potential effects of dietary mycotoxin on growth performance, haematological and serum biochemical constituents of rabbit, 24 mature female rabbits were assigned to three diets containing serum biochemical constituents of rabbit, 24 mature female rabbits were assigned to three diets containing 0, 5.0 and 10.0 mg fumonisin/kg
0, 5.0 and 10.0 mg fumonisin/kg constituting the control diet, and diets 1 and 2, respectively. The animals constituting the control diet, and diets 1 and 2, respectively. The animals were initially maintained on these diets for 2 weeks before they were mated and subsequently for 4 weeks were initially maintained on these diets for 2 weeks before they were mated and subsequently for 4 weeks after mating. Dietary fumonisin signifi cantly (P<0.05) reduced the daily dry matter intake (DMI) and fi nal after mating. Dietary fumonisin signifi cantly (P<0.05) reduced the daily dry matter intake (DMI) and fi nal live weight. The daily DMI of the rabbits fed diets 1 and 2 were 6.32 and 50.13% respectively lower than the live weight. The daily DMI of the rabbits fed diets 1 and 2 were 6.32 and 50.13% respectively lower than the daily DMI of rabbits fed the control diet. The erythrocyte counts, packed cell volume and haemoglobin values daily DMI of rabbits fed the control diet. The erythrocyte counts, packed cell volume and haemoglobin values signifi cantly (P<0.05) decreased, while the leukocyte values of the pregnant rabbits increased with the increase signifi cantly (P<0.05) decreased, while the leukocyte values of the pregnant rabbits increased with the increase in the dietary fumonisin concentrations. Pregnant does fed the control diet had signifi cantly (P<0.05) higher in the dietary fumonisin concentrations. Pregnant does fed the control diet had signifi cantly (P<0.05) higher serum total protein concentrations than those fed diets 1 and 2. The serum enzymes signifi cantly (P<0.05) serum total protein concentrations than those fed diets 1 and 2. The serum enzymes signifi cantly (P<0.05) increased in rabbits fed diets 1 and 2. Diet containing 5 mg fumonisin/kg may reduce growth performance and increased in rabbits fed diets 1 and 2. Diet containing 5 mg fumonisin/kg may reduce growth performance and induce negative responses in the haematology and serum biochemistry of pregnant does, which may affect the induce negative responses in the haematology and serum biochemistry of pregnant does, which may affect the proper development of foetuses.
proper development of foetuses.
Key words:
Key words: Fusarium verticillioidesFusarium verticillioides, fumonisin, haematology, pregnant rabbit, serum biochemistry , fumonisin, haematology, pregnant rabbit, serum biochemistry
Introduction Introduction
The contamination of feeds and foodstuffs with mycotoxins has presented a hazard The contamination of feeds and foodstuffs with mycotoxins has presented a hazard to human and animal health for decades, and this threat can only become more important to human and animal health for decades, and this threat can only become more important as the demand on the available food supply increases with the increase in population as the demand on the available food supply increases with the increase in population ((NELSON et al., 1993NELSON et al., 1993). If the food supply is limited, the mycotoxin hazards increase since ). If the food supply is limited, the mycotoxin hazards increase since more fungus-damaged, potentially mycotoxin-containing foodstuffs are consumed rather more fungus-damaged, potentially mycotoxin-containing foodstuffs are consumed rather **Corresponding authorCorresponding author::
Dr. Francis A. Gbore, School of Biological Sciences, Universiti Sains Malaysia, 11800 USM, Pulau Pinang, Malaysia, E-mail: Dr. Francis A. Gbore, School of Biological Sciences, Universiti Sains Malaysia, 11800 USM, Pulau Pinang, Malaysia, E-mail: fgbore@yahoo.co
than discarded, and malnutrition has been reported (
than discarded, and malnutrition has been reported (NELSON et al., 1993NELSON et al., 1993) to enhance the ) to enhance the susceptibility to lower concentrations of food-borne mycotoxins.
susceptibility to lower concentrations of food-borne mycotoxins.
Mycotoxins have been reported to be involved in a chain of metabolic disorders Mycotoxins have been reported to be involved in a chain of metabolic disorders in animals (
in animals (RILEY et al., 1996RILEY et al., 1996). Among such mycotoxins of signi). Among such mycotoxins of signififi cant health concern cant health concern to both man and animals is fumonisin, produced by
to both man and animals is fumonisin, produced by Fusarium verticillioides Fusarium verticillioides (Sacc.) (Sacc.)
Nirenberg, which grows on any nourishing medium (
Nirenberg, which grows on any nourishing medium (PETZINGER and WEIDENBACH, PETZINGER and WEIDENBACH, 2002
2002). Mycotoxins may cause gastrointestinal problems (). Mycotoxins may cause gastrointestinal problems (GBORE and EGBUNIKE, 2007GBORE and EGBUNIKE, 2007), ), immune suppression (
immune suppression (RICHARD et al., 1991RICHARD et al., 1991), reproductive organ problems (), reproductive organ problems (SZILÁGYI et SZILÁGYI et al., 1994; OGUNLADE et al., 2006
al., 1994; OGUNLADE et al., 2006), blood abnormalities (), blood abnormalities (EWUOLA and EGBUNIKE, 2008; EWUOLA and EGBUNIKE, 2008; EWUOLA et al., 2008; GBORE and EGBUNIKE, 2008a, 2009
EWUOLA et al., 2008; GBORE and EGBUNIKE, 2008a, 2009), disturbances in the immune ), disturbances in the immune system (
system (VOSS et al., 2007VOSS et al., 2007), suppression in sperm production and reproductive performance ), suppression in sperm production and reproductive performance ((GBORE and EGBUNIKE, 2008b; GBORE, 2009aGBORE and EGBUNIKE, 2008b; GBORE, 2009a), and delayed sexual maturity (), and delayed sexual maturity (GBORE, GBORE, 2009b
2009b). ).
Blood contains a myriad of metabolites and other constituents, which provide a Blood contains a myriad of metabolites and other constituents, which provide a valuable medium for clinical investigation and assessment of nutritional status of human valuable medium for clinical investigation and assessment of nutritional status of human beings and animals. Dietary components have measurable effects on blood components; beings and animals. Dietary components have measurable effects on blood components; hence, blood constituents are widely used in nutritional evaluation and survey of animals hence, blood constituents are widely used in nutritional evaluation and survey of animals ((OLORODE et al., 1995OLORODE et al., 1995). ).
Based on the reviewed physiological effects of dietary fumonisin on animals, coupled Based on the reviewed physiological effects of dietary fumonisin on animals, coupled with a survey of contemporary literature revealing an increasing wave of fumonisin with a survey of contemporary literature revealing an increasing wave of fumonisin contamination of feeds and feedstuff (
contamination of feeds and feedstuff (FAZEKAS et al., 1997FAZEKAS et al., 1997)),, this study was designed this study was designed
to assess the growth performance, haematological and serum biochemical profi les of to assess the growth performance, haematological and serum biochemical profi les of pregnant rabbits fed diets containing
pregnant rabbits fed diets containing FF.. verticillioides verticillioides-contaminated maize. -contaminated maize.
Materials and methods Materials and methods
Fumonisin production and experimental diets
Fumonisin production and experimental diets.. Autoclaved maize grains were Autoclaved maize grains were
cultured with a toxigenic strain of
cultured with a toxigenic strain of F. verticillioidesF. verticillioides (MRC 286) inoculum obtained from (MRC 286) inoculum obtained from
the Plant Pathology Laboratory of the International Institute of Tropical Agriculture the Plant Pathology Laboratory of the International Institute of Tropical Agriculture (IITA), Ibadan, Nigeria to produce fumonisin as described previously (
(IITA), Ibadan, Nigeria to produce fumonisin as described previously (NELSON et al., NELSON et al., 1994
1994). Brie). Brieflfl y, the method involved the addition of 500 g of yellow corn kernels and y, the method involved the addition of 500 g of yellow corn kernels and 500 mL of distilled water in a 30.5×61cm autoclavable polyethylene bag. The corn was 500 mL of distilled water in a 30.5×61cm autoclavable polyethylene bag. The corn was inoculated by drawing an aqueous suspension from a lyophilized culture into a sterile 5 inoculated by drawing an aqueous suspension from a lyophilized culture into a sterile 5 mL syringe fi tted with a 19-gauge needle and injecting 1 mL through the side of each mL syringe fi tted with a 19-gauge needle and injecting 1 mL through the side of each bag. Bags of inoculated corn were incubated in the dark at 20 to 22
bag. Bags of inoculated corn were incubated in the dark at 20 to 22 ooC for four weeks. C for four weeks.
Seven to eight days after inoculation, holes were punched near the tops of the bags to Seven to eight days after inoculation, holes were punched near the tops of the bags to promote aeration. After a four-week incubation period, the culture material was air dried promote aeration. After a four-week incubation period, the culture material was air dried for 48 hours.
for 48 hours. Uncontaminated maize grains and the Uncontaminated maize grains and the FusariumFusarium-contaminated maize grains -contaminated maize grains
were used to formulate three diets. Dietary contents of common
(afl atoxin),
(afl atoxin), FusariumFusarium mycotoxins including deoxynivalenol (DON, vomitoxin), T-2 mycotoxins including deoxynivalenol (DON, vomitoxin), T-2
toxin, zearalenone, and fumonisin were analysed using mycotoxin quantitative toxin, zearalenone, and fumonisin were analysed using mycotoxin quantitative CD-ELISA test kits (Neogen, Lansing, MI, USA). The concentrations of fumonisin in the ELISA test kits (Neogen, Lansing, MI, USA). The concentrations of fumonisin in the diets were approximately 0, 5 and 10 mg/kg constituting the control diet (which had no diets were approximately 0, 5 and 10 mg/kg constituting the control diet (which had no
Fusarium
Fusarium-contaminated maize grains), diets 1 (medium -contaminated maize grains), diets 1 (medium FusariumFusarium-infected diet) and 2 -infected diet) and 2
(high
(high FusariumFusarium-infected diet), respectively. All other mycotoxins screened were below -infected diet), respectively. All other mycotoxins screened were below
the 0.2 mg/kg detection limit. the 0.2 mg/kg detection limit.
Experimental animals and treatment.
Experimental animals and treatment. Twenty-four clinically normal matured Twenty-four clinically normal matured
crossbred female rabbits weighing between 1.65 and 2.0 kg sourced from the Animal crossbred female rabbits weighing between 1.65 and 2.0 kg sourced from the Animal Science Unit of the Federal College of Agriculture Farm, Akure, Ondo State, Nigeria, Science Unit of the Federal College of Agriculture Farm, Akure, Ondo State, Nigeria, were randomly assigned into one of the three diets (eight per treatment) in a six-week were randomly assigned into one of the three diets (eight per treatment) in a six-week feeding trial. The animals were maintained on the diets for 2 weeks before they were feeding trial. The animals were maintained on the diets for 2 weeks before they were mated and subsequently for 4 weeks after mating. The composition of the experimental mated and subsequently for 4 weeks after mating. The composition of the experimental diets is shown in Table 1.
diets is shown in Table 1.
Data collection and analysis.
Data collection and analysis. Feed consumption for each animal was measured daily Feed consumption for each animal was measured daily
by the difference between the daily feed supplied and refusal, and live-weight changes by the difference between the daily feed supplied and refusal, and live-weight changes of the animals were taken weekly throughout the experimental period. At the end of the of the animals were taken weekly throughout the experimental period. At the end of the feeding experiment, a blood sample was collected from the jugular vein of each animal into feeding experiment, a blood sample was collected from the jugular vein of each animal into two sets of Monoject
two sets of Monoject®® vacutainers. One set containing Ethylene diaminetetraacetic acid vacutainers. One set containing Ethylene diaminetetraacetic acid
(EDTA) for haematology, while the other set without EDTA was covered and centrifuged, (EDTA) for haematology, while the other set without EDTA was covered and centrifuged, the serum decanted and deep-frozen for serum biochemical and enzymological analyses. the serum decanted and deep-frozen for serum biochemical and enzymological analyses. The erythrocyte, the total leukocyte and the differential leukocyte counts, the packed The erythrocyte, the total leukocyte and the differential leukocyte counts, the packed cell volume (PCV) and haemoglobin (Hb) concentrations were determined as described cell volume (PCV) and haemoglobin (Hb) concentrations were determined as described by
by EWUOLA and EGBUNIKE (2008) EWUOLA and EGBUNIKE (2008), the blood corpuscular constants: mean cell volume , the blood corpuscular constants: mean cell volume (MCV), mean cell haemoglobin (MCH) and mean cell haemoglobin concentration (MCV), mean cell haemoglobin (MCH) and mean cell haemoglobin concentration (MCHC) were determined using appropriate formulae as described by
(MCHC) were determined using appropriate formulae as described by JAIN (1986)JAIN (1986). . The serum chloride (Cl
The serum chloride (Cl--), creatinine, and urea nitrogen were estimated as described ), creatinine, and urea nitrogen were estimated as described
previously (
previously (GBORE et al., 2006GBORE et al., 2006). The routine ). The routine flfl ame photometric technique of ame photometric technique of VARLEY et VARLEY et al. (1980)
al. (1980) was used to determine the serum levels of sodium (Na was used to determine the serum levels of sodium (Na++) and potassium (K) and potassium (K++). ).
The serum phosphate (PO
The serum phosphate (PO443-3-) was determined spectrometrically as described by) was determined spectrometrically as described by GARBER GARBER
and MILLER (1983)
and MILLER (1983), the serum bicarbonate (HCO, the serum bicarbonate (HCO33--) was determined by titration; while ) was determined by titration; while
the serum calcium (C
the serum calcium (Ca2+) was determined by atomic absorption photometry as outlined ) was determined by atomic absorption photometry as outlined
by
by TRUDEAN and FREIER (1967)TRUDEAN and FREIER (1967). The serum enzymes: alanine aminotransferase (ALT), . The serum enzymes: alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP), were evaluated using aspartate aminotransferase (AST) and alkaline phosphatase (ALP), were evaluated using a commercial test kit (Randox Laboratories Ltd., Crumlin, Co. Antrim, UK).
a commercial test kit (Randox Laboratories Ltd., Crumlin, Co. Antrim, UK).
All the data obtained were subjected to statistical analysis using ANOVA procedure All the data obtained were subjected to statistical analysis using ANOVA procedure of
of SAS (1999)SAS (1999). The Duncan’s multiple range test of the same software was used to separate . The Duncan’s multiple range test of the same software was used to separate all means at 5% probability level.
Results Results
The daily dry matter intake (DMI) and weight gain of female rabbits fed varying The daily dry matter intake (DMI) and weight gain of female rabbits fed varying levels of dietary fumonisin are shown in Table 2. The DMI and fi nal live weights of the levels of dietary fumonisin are shown in Table 2. The DMI and fi nal live weights of the animals were signifi cantly (P<0.05) infl uenced. The DMI of the rabbits fed diets 1 and animals were signifi cantly (P<0.05) infl uenced. The DMI of the rabbits fed diets 1 and 2 were 6.32 and 50.13% respectively lower than the daily feed intake of rabbits fed the 2 were 6.32 and 50.13% respectively lower than the daily feed intake of rabbits fed the control diet. Similarly, the weight gains of the rabbits fed diets 1 and 2 were 79.67 and control diet. Similarly, the weight gains of the rabbits fed diets 1 and 2 were 79.67 and 76.92% of the weight gained by rabbits fed the control diet.
76.92% of the weight gained by rabbits fed the control diet.
Table 1. Gross composition (%) of the test diets Table 1. Gross composition (%) of the test diets Dietary fumonisin level
Ingredient Control (0 mg/kg) Diet 1 (5 mg/kg) Diet 2 (10 mg/kg)
Non-inoculated maize 25.00 20.00 15.00
Inoculated maize* - 5.00 10.00
Rice husk 30.00 30.00 30.00
Wheat offal 25.00 25.00 25.00
Palm kernel cake 10.00 10.00 10.00
Soybeans meal 5.00 5.00 5.00 Fishmeal 2.00 2.00 2.00 Dicalcium phosphate 2.00 2.00 2.00 Salt 0.50 0.50 0.50 Minerals/vitamin premix** 0.45 0.45 0.45 Methionine 0.03 0.03 0.03 Lysine 0.02 0.02 0.02 Calculated nutrients Crude protein (%) 13.15 13.15 13.15 Crude fi bre (%) 13.97 13.97 13.97
Digestible energy (kcal/kg) 2225.21 2225.21 2225.21
*Inoculated with
*Inoculated with F. verticillioidesF. verticillioides. ** To provide per kg of diet: vitamin A (10,000 i.u.), vitamin D (20,000 i.u.), . ** To provide per kg of diet: vitamin A (10,000 i.u.), vitamin D (20,000 i.u.), vitamin E (5 i.u.), vitamin K (2.5 mg), choline (350 mg), folic acid (1 mg), manganese (56 mg), iodine (1 mg), vitamin E (5 i.u.), vitamin K (2.5 mg), choline (350 mg), folic acid (1 mg), manganese (56 mg), iodine (1 mg), iron (20 mg), copper (10 mg), zinc (50 mg), cobalt (1.25 mg).
iron (20 mg), copper (10 mg), zinc (50 mg), cobalt (1.25 mg).
Table 2. Performance of pregnant does fed dietary fumonisin (Mean Table 2. Performance of pregnant does fed dietary fumonisin (Mean ±± SE) SE)
Dietary fumonisin level
Parameter Control (0 mg/kg) Diet 1 (5 mg/kg) Diet 2 (10 mg/kg)
Initial live weight (kg) 1.79 ± 0.04 1.88 ± 0.55 1.73 ± 0.03
Final live weight (kg) 1.82 ± 0.05a 1.45 ± 0.38b 1.40 ± 0.04c
Dry matter intake (g) 41.15 ± 2.59a 38.55 ± 10.23b 20.57 ± 9.49b
abc:
Table 3. Haematological parameters of pregnant does fed dietary fumonisin (Mean Table 3. Haematological parameters of pregnant does fed dietary fumonisin (Mean ±± SE) SE)
Dietary fumonisin level
Parameter Control (0 mg/kg) Diet 1 (5 mg/kg) Diet 2 (10 mg/kg)
Packed cell volume (%) 36.67 ± 1.33a 33.00 ± 4.00a 20.33 ± 1.45b
Haemoglobin (g/L) 139.67 ± 2.19a 119.00 ± 7.00b 87.67 ± 5.21c
Erythrocyte (×1012/L) 5.60 ± 1.07a 3.80 ± 0.70ab 2.30 ± 18.95b
Mean cell volume (fL) 69.74 ± 11.21 87.89 ± 5.66 89.29 ± 5.46
Mean cell Haemoglobin (ug) 269.21 ± 52.23 320.65 ± 40.65 385.24 ± 20.19
MCHC* (%) 38.18 ± 1.32 36.34 ± 2.28 43.34 ± 2.62 Leucocytes (×109/L) 7.80 ± 1.77a 12.05 ± 1.75b 12.63 ± 1.69b Neutrophils (%) 38.67 ± 4.10b 61.50 ± 13.50a 47.67 ± 1.20ab Lymphocytes (%) 60.00 ± 4.04a 36.50 ± 13.50b 50.33 ± 1.76ab Monocytes (%) 0.33 ± 0.33 0.50 ± 0.50 0.67 ± 0.33 Eosinophils (%) 1.00 ± 0.58 1.50 ± 0.50 1.33 ± 0.33
*MCHC: - Mean cell haemoglobin concentration.
*MCHC: - Mean cell haemoglobin concentration. abcabc: Means within the same row with different superscripts : Means within the same row with different superscripts differ signifi cantly (P<0.05).
differ signifi cantly (P<0.05).
Table 4. Serum proteins and enzymes of pregnant does fed dietary fumonisin (Mean Table 4. Serum proteins and enzymes of pregnant does fed dietary fumonisin (Mean ±± SE) SE)
Dietary fumonisin level
Parameter Control (0 mg/kg) Diet 1 (5 mg/kg) Diet 2 (10 mg/kg)
Total protein (g/L) 80.00 ± 4.08a 70.00 ± 0.09b 65.00 ± 2.04b Albumin (g/L) 51.50 ± 5.10a 47.50 ± 2.65ab 38.50 ± 1.43b Globulin (g/L) 28.50 ± 1.02 22.50 ± 2.65 26.50 ± 3.47 Albumin/globulin 1.81 ± 0.24 2.11 ± 0.40 1.45 ± 0.27 ALT* (U/L) 55.50 ± 1.38b 100.00 ± 1.23a 122.50 ± 1.02 a AST** (U/L) 46.25 ± 0.62c 108.25 ± 0.72b 125.00 ± 2.04a ALP*** (U/L) 11.05 ± 0.14b 12.40 ± 0.14b 18.52 ± 0.16a *Alanine aminot
*Alanine aminotransferase, **Aspartate aminotransferase, *** Alkaline phosphatase. ransferase, **Aspartate aminotransferase, *** Alkaline phosphatase. abcabc: Means on the same : Means on the same row with different
Table 5. Serum electrolytes of pregnant does fed dietary fumonisin (Mean Table 5. Serum electrolytes of pregnant does fed dietary fumonisin (Mean ±± SE) SE)
Dietary fumonisin level
Parameter Control (0 mg/kg) Diet 1 (5 mg/kg) Diet 2 (10 mg/kg)
Calcium (mmol/L) 12.60 ± 0.33 12.10 ± 0.86 11.65 ± 0.31 Phosphate (mmol/L) 4.30 ± 0.20b 7.50 ± 0.45a 7.60 ± 0.08a Sodium (mmol/L) 132.0 ± 01.63 129.00 ± 0.41 131.00 ± 2.04 Potassium (mmol/L) 4.05 ± 0.06a 3.85 ± 0.06a 4.00 ± 0.04ab Chloride (mmol/L) 108.75 ± 0.51ab 117.50 ± 3.06a 100.00 ± 4.08b Bicarbonate (mmol/L) 24.50 ± 0.61c 27.50 ± 0.20a 25.75 ± 0.10b
Urea nitrogen (mmol/L) 16.50 ± 0.60 19.50 ± 0.43 19.50 ± 0.44
Creatinine (μmol/L) 1.60 ± 0.02b 1.90 ± 0.05a 1.70 ± 0.04ab
abc
abc: Means on the same row with different superscripts differ signi: Means on the same row with different superscripts differ signififi cantly (P<0.05) cantly (P<0.05)
The haematological values of pregnant rabbits fed varied levels of dietary fumonisin The haematological values of pregnant rabbits fed varied levels of dietary fumonisin are shown in Table 3. The mean values of the erythrocyte counts, PCV and Hb signifi cantly are shown in Table 3. The mean values of the erythrocyte counts, PCV and Hb signifi cantly (P<0.05) decreased with the increase in the dietary fumonisin concentrations, while the (P<0.05) decreased with the increase in the dietary fumonisin concentrations, while the leukocyte values of the pregnant rabbits increased. The PCV of the pregnant rabbits fed leukocyte values of the pregnant rabbits increased. The PCV of the pregnant rabbits fed diet 2 was signifi cantly (P<0.05) lower than those fed diet 1 and the control diet, while the diet 2 was signifi cantly (P<0.05) lower than those fed diet 1 and the control diet, while the erythrocyte count of the rabbits fed diet 2, which was signifi cantly (P<0.05) lower than the erythrocyte count of the rabbits fed diet 2, which was signifi cantly (P<0.05) lower than the erythrocyte counts of those fed the control diet, was not statistically different from those erythrocyte counts of those fed the control diet, was not statistically different from those fed diet 1. The mean leukocyte count of the rabbits fed diet 2 was signifi cantly (P<0.05) fed diet 1. The mean leukocyte count of the rabbits fed diet 2 was signifi cantly (P<0.05) higher than those fed the control diet but not different statistically from those fed diet higher than those fed the control diet but not different statistically from those fed diet 1. The neutrophils increased signifi cantly (P<0.05) while the lymphocytes signifi cantly 1. The neutrophils increased signifi cantly (P<0.05) while the lymphocytes signifi cantly (P<0.05) decreased with the increase in the dietary fumonisin concentrations.
(P<0.05) decreased with the increase in the dietary fumonisin concentrations.
Table 4 shows the serum proteins and enzyme activities of pregnant rabbits fed varied Table 4 shows the serum proteins and enzyme activities of pregnant rabbits fed varied dietary fumonisin levels. Pregnant does fed the control diet had signifi cantly (P<0.05) dietary fumonisin levels. Pregnant does fed the control diet had signifi cantly (P<0.05) higher mean serum total protein concentrations than those on diets 1 and 2. The serum higher mean serum total protein concentrations than those on diets 1 and 2. The serum total protein concentrations of the rabbits on diets 1 and 2, respectively, were about 12.5 total protein concentrations of the rabbits on diets 1 and 2, respectively, were about 12.5 and 19% lower than those on the control diet. The mean serum albumin concentration and 19% lower than those on the control diet. The mean serum albumin concentration of the pregnant rabbits fed diet 2 was signifi cantly (P<0.05) lower than the mean serum of the pregnant rabbits fed diet 2 was signifi cantly (P<0.05) lower than the mean serum albumin of those fed the control diet, which was not signifi cantly (P>0.05) higher than the albumin of those fed the control diet, which was not signifi cantly (P>0.05) higher than the serum albumin of those on diet 1. The serum ALT and AST values of the rabbits fed the serum albumin of those on diet 1. The serum ALT and AST values of the rabbits fed the control diet were signifi cantly (P<0.05) lower than the serum ALT and AST of those fed control diet were signifi cantly (P<0.05) lower than the serum ALT and AST of those fed diets 1 and 2, while the serum ALP activities of the rabbits fed the control diet and diet 1 diets 1 and 2, while the serum ALP activities of the rabbits fed the control diet and diet 1 were signifi cantly (P<0.05) lower than those on diet 2.
The serum phosphate, bicarbonate and creatinine values signifi cantly (P<0.05) The serum phosphate, bicarbonate and creatinine values signifi cantly (P<0.05) increased, while the serum potassium and chloride values decreased with increase in the increased, while the serum potassium and chloride values decreased with increase in the dietary fumonisin (Table 5).
dietary fumonisin (Table 5). Discussion
Discussion
The United States Federal Department of Agriculture (
The United States Federal Department of Agriculture (ANONYM., 2001aANONYM., 2001a) considered ) considered rabbits, grouped with horses, as the most sensitive species to fumonisin in animal feeds. rabbits, grouped with horses, as the most sensitive species to fumonisin in animal feeds. In this study, since all the animals were fed
In this study, since all the animals were fed ad libitumad libitum, the drastic decline in the DMI and , the drastic decline in the DMI and
the relative change in the body weight of rabbits fed diets 1 and 2 may be attributed to the relative change in the body weight of rabbits fed diets 1 and 2 may be attributed to the adverse effects of the mycotoxin on feed intake and nutrient utilization as observed the adverse effects of the mycotoxin on feed intake and nutrient utilization as observed in pigs fed dietary fumonisin by
in pigs fed dietary fumonisin by GBORE and EGBUNIKE (2007)GBORE and EGBUNIKE (2007). As all the animals were . As all the animals were fed
fed ad libitumad libitum and all the diets were isocaloric and isonitrogenous, the concentration- and all the diets were isocaloric and isonitrogenous, the
concentration-dependent decline in fi nal live weight and feed intake indicates the role that fumonisin dependent decline in fi nal live weight and feed intake indicates the role that fumonisin can play in animal nutrition and subsequent growth performance. These results are in can play in animal nutrition and subsequent growth performance. These results are in agreement with results from similar studies that dietary fumonisin depressed live weight agreement with results from similar studies that dietary fumonisin depressed live weight gain in animals.
gain in animals. GELDERBLOM GELDERBLOM et al. (1988)et al. (1988) reported that the mean body weights of BD reported that the mean body weights of BD IX rats, consuming a diet containing 1 g fumonisin B
IX rats, consuming a diet containing 1 g fumonisin B11/kg during a 4-week promotion /kg during a 4-week promotion treatment were 50% lower than those of non-treated rats. Also, US NTP (
treatment were 50% lower than those of non-treated rats. Also, US NTP (ANONYM., ANONYM., 2001b)
2001b) reported less gain in body weight in rats exposed to fumonisin-containing diet at reported less gain in body weight in rats exposed to fumonisin-containing diet at 56 mg fumonisin B
56 mg fumonisin B11/kg body weight after 28 days of feeding in a range-/kg body weight after 28 days of feeding in a range-fifi nding study. nding study. Haematological indices are an index and a refl ection of the effects of dietary treatments Haematological indices are an index and a refl ection of the effects of dietary treatments on animals in terms of the quality of feed ingested and nutrients available to an animal on animals in terms of the quality of feed ingested and nutrients available to an animal to meet its physiological requirements. In this present study, the signifi cantly reduced to meet its physiological requirements. In this present study, the signifi cantly reduced erythrocyte and PCV values were inversely related to the fumonisin concentrations in the erythrocyte and PCV values were inversely related to the fumonisin concentrations in the diets, with the animals that consumed the diet containing the higher dietary mycotoxin diets, with the animals that consumed the diet containing the higher dietary mycotoxin concentrations recording the lowest values for both parameters. The mean values of the concentrations recording the lowest values for both parameters. The mean values of the erythrocytes and PCV of pregnant does on diets 1 and 2 fed varied levels of dietary erythrocytes and PCV of pregnant does on diets 1 and 2 fed varied levels of dietary fumonisin were not within the normal physiological ranges of 5.11 - 6.51
fumonisin were not within the normal physiological ranges of 5.11 - 6.51×10101212/L and 31.0 /L and 31.0
- 48.6% reported by
- 48.6% reported by MITRUKA and RAWNSLEY (1977)MITRUKA and RAWNSLEY (1977) for young adult female rabbits. for young adult female rabbits. The values of haemoglobin, an iron-containing conjugated protein that performs the The values of haemoglobin, an iron-containing conjugated protein that performs the physiological function of transporting oxygen and carbon dioxide, which signifi cantly physiological function of transporting oxygen and carbon dioxide, which signifi cantly decreased in rabbits fed diets 1 and 2 compared with those on the control diet, suggest decreased in rabbits fed diets 1 and 2 compared with those on the control diet, suggest that the animals on diets 1 and 2 suffered depressed respiratory capability. The results also that the animals on diets 1 and 2 suffered depressed respiratory capability. The results also revealed that the rabbits exposed to diets containing
revealed that the rabbits exposed to diets containing FusariumFusarium-inoculated maize (diets -inoculated maize (diets
1 and 2) suffered signifi cantly from the synthesis (erythropoiesis) and concentration of 1 and 2) suffered signifi cantly from the synthesis (erythropoiesis) and concentration of erythrocytes. The corresponding statistical decrease in the PCV of the animals exposed to erythrocytes. The corresponding statistical decrease in the PCV of the animals exposed to diets 1 and 2 revealed that the animals were anaemic.
The circulating leukocyte counts of the animals fed diets 1 and 2 were above the normal The circulating leukocyte counts of the animals fed diets 1 and 2 were above the normal physiological range of 5.20 - 10.6
physiological range of 5.20 - 10.6×10109/L for leukocyte counts of young adult female rabbits /L for leukocyte counts of young adult female rabbits
reported by
reported by MITRUKA and RAWNSLEY (1977)MITRUKA and RAWNSLEY (1977). The results indicate that the animals fed . The results indicate that the animals fed diets 1 and 2 might have suffered leukocytosis. According to
diets 1 and 2 might have suffered leukocytosis. According to COLES (1986)COLES (1986), leukocytosis , leukocytosis may result from intoxications including those produced by metabolic disturbance. The may result from intoxications including those produced by metabolic disturbance. The signifi cant effects of dietary fumonisin on haematological parameters observed in this signifi cant effects of dietary fumonisin on haematological parameters observed in this study were in contrast to the report of
study were in contrast to the report of OGUNLADE et alOGUNLADE et al.. (2004) (2004). Also, . Also, PARENT-MASSIN PARENT-MASSIN and PARCHMENT (1998)
and PARCHMENT (1998) and and ZOMBORSZKY-KOVÁCS et alZOMBORSZKY-KOVÁCS et al.. (2002) (2002) considered fumonisin considered fumonisin
as non-haematotoxic. The fi ndings in this present study however agreed with the report as non-haematotoxic. The fi ndings in this present study however agreed with the report of
of ESPADA et alESPADA et al.. (1994) (1994) that revealed altered haematological parameters in broiler chicks that revealed altered haematological parameters in broiler chicks
fed diets formulated from
fed diets formulated from F. verticillioidesF. verticillioides (MRC 826) culture containing 30 to 300mg (MRC 826) culture containing 30 to 300mg
fumonisin B
fumonisin B11/kg diet. Also, altered haematological parameters have been reported in /kg diet. Also, altered haematological parameters have been reported in minks (
minks (POWELL et al., 1996POWELL et al., 1996), rabbit bucks (), rabbit bucks (EWUOLA and EGBUNIKE, 2008EWUOLA and EGBUNIKE, 2008) and pigs ) and pigs ((ROTTER et al., 1996; GBORE and EGBUNIKE, 2008aROTTER et al., 1996; GBORE and EGBUNIKE, 2008a) fed dietary fumonisin. In one-year ) fed dietary fumonisin. In one-year old carp, consumption of nutritionally balanced pellets contaminated with 0.5 and 5.0 mg old carp, consumption of nutritionally balanced pellets contaminated with 0.5 and 5.0 mg fumonisin B
fumonisin B11 per kg body weight for 42 days resulted in alterations of haematological per kg body weight for 42 days resulted in alterations of haematological and serum biochemical parameters compared with the control (
and serum biochemical parameters compared with the control (PEPELJNJAK et al. 2003PEPELJNJAK et al. 2003). ). The authors suggested that fumonisin B
The authors suggested that fumonisin B11 probably interferes with the carp’s respiratory probably interferes with the carp’s respiratory process.
process.
Reduced tissue protein synthesis has been reported (
Reduced tissue protein synthesis has been reported (DÄNICKE et al., 2006DÄNICKE et al., 2006) to be ) to be one of the major symptoms seen in pigs consuming
one of the major symptoms seen in pigs consuming FusariumFusarium-contaminated feed. The -contaminated feed. The
signifi cantly lower levels of serum proteins in rabbits fed diets 1 and 2 in this study signifi cantly lower levels of serum proteins in rabbits fed diets 1 and 2 in this study indicated that the dietary fumonisin might have induced some alterations in protein indicated that the dietary fumonisin might have induced some alterations in protein metabolism in the animals, since serum protein synthesis is related to the amount of metabolism in the animals, since serum protein synthesis is related to the amount of available protein in the diet (
available protein in the diet (IYAYI and TEWE, 1998IYAYI and TEWE, 1998). The mycotoxin might have inhibited ). The mycotoxin might have inhibited protein metabolism in the animals as reported for sphingolipid synthesis (
protein metabolism in the animals as reported for sphingolipid synthesis (RILEY et al., RILEY et al., 1996
1996) or protein digestibility and absorption as observed by ) or protein digestibility and absorption as observed by GBORE and EGBUNIKE (2007)GBORE and EGBUNIKE (2007). . Also, the disruption of membrane cellular structure by cytotoxic mycotoxins, which Also, the disruption of membrane cellular structure by cytotoxic mycotoxins, which interferes with vital cellular processes, including protein synthesis, has been reported interferes with vital cellular processes, including protein synthesis, has been reported ((GUERRE et al., 2000GUERRE et al., 2000). ).
The decrease in total protein, globulin and albumin concentrations in the sera of The decrease in total protein, globulin and albumin concentrations in the sera of pregnant rabbits fed diets 1 and 2 suggested defi cient protein synthesis in the animals pregnant rabbits fed diets 1 and 2 suggested defi cient protein synthesis in the animals compared with the rabbits fed the control diet. The dietary mycotoxin might have elicited compared with the rabbits fed the control diet. The dietary mycotoxin might have elicited some pathological and physiological changes in the animals, leading to poor digestion, some pathological and physiological changes in the animals, leading to poor digestion, poor absorption or poor utilization of protein in the diets.
poor absorption or poor utilization of protein in the diets.
CHAMPE et al. (2007)
CHAMPE et al. (2007) stated that aminotransferases are normally intracellular enzymes, stated that aminotransferases are normally intracellular enzymes, with low levels found in the plasma representing the release of cellular contents during with low levels found in the plasma representing the release of cellular contents during normal cell turnover. The serum ALT and AST levels, according to
are elevated in nearly all liver diseases and are particularly high in conditions that cause are elevated in nearly all liver diseases and are particularly high in conditions that cause extensive cell necrosis, including severe viral hepatitis or toxic liver injury. Mean values extensive cell necrosis, including severe viral hepatitis or toxic liver injury. Mean values of ALT and AST obtained for rabbits fed diets 1 and 2 were above the reported ranges of of ALT and AST obtained for rabbits fed diets 1 and 2 were above the reported ranges of 48.5-78.9 U/L and 42.5-98.0 U/L for normal young adult female rabbits (
48.5-78.9 U/L and 42.5-98.0 U/L for normal young adult female rabbits (MITRUKA and MITRUKA and RAWNSLEY, 1977
RAWNSLEY, 1977) for the respective serum enzymes. However, only the ALP value of ) for the respective serum enzymes. However, only the ALP value of 18.52 i.u/l obtained for the female rabbits on diet 2 was above the physiological reference 18.52 i.u/l obtained for the female rabbits on diet 2 was above the physiological reference range reported by
range reported by MITRUKA and RAWNSLEY (1977) MITRUKA and RAWNSLEY (1977). The serum enzyme activities above . The serum enzyme activities above the normal ranges are abnormal and are an indication that the animals might have suffered the normal ranges are abnormal and are an indication that the animals might have suffered liver and/or kidney damage. Signifi cant increases in the serum activities of these enzymes, liver and/or kidney damage. Signifi cant increases in the serum activities of these enzymes, indicating damage to the liver and kidney, were reported by
indicating damage to the liver and kidney, were reported by VOSS et al. (1993)VOSS et al. (1993) in male and in male and female Sprague-Dawley rats fed diets containing 0-150 mg of fumonisin B
female Sprague-Dawley rats fed diets containing 0-150 mg of fumonisin B11 for 4 weeks. for 4 weeks.
RESTUM et al. (1995)
RESTUM et al. (1995) reported higher serum ALT, AST and ALP activities in minks fed reported higher serum ALT, AST and ALP activities in minks fed 119 ppm fumonisin B
119 ppm fumonisin B11 diet as compared to the control. Similar evidence of abnormal diet as compared to the control. Similar evidence of abnormal increases in serum enzyme activities, probably due to cellular destruction induced by increases in serum enzyme activities, probably due to cellular destruction induced by fumonisin, has been reported (
fumonisin, has been reported (GELDERBLOM et al., 1994; MEHTA et al., 1998; EWUOLA and GELDERBLOM et al., 1994; MEHTA et al., 1998; EWUOLA and EGBUNIKE, 2008
EGBUNIKE, 2008). Recently, signi). Recently, signififi cant increases in serum enzymes, urea and creatinine cant increases in serum enzymes, urea and creatinine were reported in rabbits administered a single oral dose of 31.5 mg fumonisin B
were reported in rabbits administered a single oral dose of 31.5 mg fumonisin B11/kg body /kg body weight by
weight by ORSI et al. (2009)ORSI et al. (2009), a , a fifi nding the authors described as characterizing hepatic and nding the authors described as characterizing hepatic and renal injury.
renal injury.
The control of intracellular calcium (seemingly via sphingosine 1-phosphate) and The control of intracellular calcium (seemingly via sphingosine 1-phosphate) and control of plasma membrane potassium permeability in myocytes are examples of cellular control of plasma membrane potassium permeability in myocytes are examples of cellular regulatory processes that have been reported by
regulatory processes that have been reported by ANONYM. (2000)ANONYM. (2000) to be modulated by to be modulated by de de novo
novo ceramide biosynthesis as shown by inhibition of the process by fumonisin. The ceramide biosynthesis as shown by inhibition of the process by fumonisin. The
signifi cantly higher values of the serum phosphate obtained for the pregnant rabbits on signifi cantly higher values of the serum phosphate obtained for the pregnant rabbits on diets 1 and 2 were above the serum phosphate reference range of 2.30-6.90 mmol/L for diets 1 and 2 were above the serum phosphate reference range of 2.30-6.90 mmol/L for normal young adult female rabbits reported by
normal young adult female rabbits reported by MITRUKA and RAWNSLEY (1977)MITRUKA and RAWNSLEY (1977). This . This
fi nding is signifi cant as this may alter the normal calcium-phosphate ratio for proper foetal
fi nding is signifi cant as this may alter the normal calcium-phosphate ratio for proper foetal growth and development in pregnant rabbits fed diets 1 and 2. Incidences of increased growth and development in pregnant rabbits fed diets 1 and 2. Incidences of increased foetal deaths, hydrocephalic foetuses and skeletal anomalies, in addition to decreased foetal deaths, hydrocephalic foetuses and skeletal anomalies, in addition to decreased foetal length and foetal weight were observed by
foetal length and foetal weight were observed by COLLINS et al. (1998)COLLINS et al. (1998) in pregnant rats in pregnant rats given 25 and 50 mg/kg bw purifi ed fumonisin B
given 25 and 50 mg/kg bw purifi ed fumonisin B11. The serum phosphate, bicarbonate . The serum phosphate, bicarbonate and creatinine values, which increased with increase in the dietary fumonisin levels, and creatinine values, which increased with increase in the dietary fumonisin levels, agreed with the results of
agreed with the results of MOTELIN et al. (1994)MOTELIN et al. (1994) who reported dose-dependent increases who reported dose-dependent increases in biochemical parameters measured in pigs fed diets containing total fumonisins of over in biochemical parameters measured in pigs fed diets containing total fumonisins of over 5 mg/kg for 14 days. However, the signifi cant infl uence of dietary fumonisin levels on 5 mg/kg for 14 days. However, the signifi cant infl uence of dietary fumonisin levels on the serum electrolytes in this study was at variance with the report of
the serum electrolytes in this study was at variance with the report of GBORE et al. (2006) GBORE et al. (2006)
that observed no signifi cant difference in serum electrolytes of male rabbits fed diets that observed no signifi cant difference in serum electrolytes of male rabbits fed diets containing 12.30-24.60 mg fumonisin/kg for fi ve weeks. The variability in the effects of containing 12.30-24.60 mg fumonisin/kg for fi ve weeks. The variability in the effects of dietary fumonisin levels on the serum electrolytes observed by various authors might be dietary fumonisin levels on the serum electrolytes observed by various authors might be
the result of the physiological state of the animals as the developing embryos in pregnant the result of the physiological state of the animals as the developing embryos in pregnant rabbits would place more demand for these electrolytes, the doses of the toxin used or the rabbits would place more demand for these electrolytes, the doses of the toxin used or the length of exposure of the animals to the mycotoxin.
length of exposure of the animals to the mycotoxin.
This study has demonstrated that the exposure of pregnant does to diets containing This study has demonstrated that the exposure of pregnant does to diets containing 5 mg fumonisin/kg may result in signifi cantly reduced growth performance and altered 5 mg fumonisin/kg may result in signifi cantly reduced growth performance and altered haematological and serum biochemical parameters, which may negatively infl uence the haematological and serum biochemical parameters, which may negatively infl uence the proper growth and development of foetuses in pregnant rabbits.
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SAŽETAK SAŽETAK
Radi istraživanja mogućih učinaka mikotoksina u hrani na rast, hematološke i biokemijske pokazatelje Radi istraživanja mogućih učinaka mikotoksina u hrani na rast, hematološke i biokemijske pokazatelje u serumu, 24 odrasle kunice dobivale su hranu koja nije sadržavala fumonizin i bila je kontrolna, te hranu s u serumu, 24 odrasle kunice dobivale su hranu koja nije sadržavala fumonizin i bila je kontrolna, te hranu s oznakom 1 koja je sadržavala 5,0 i hranu s oznakom 2 koja je sadržavala 10,0 mg fumonizina/kg. Kunice su oznakom 1 koja je sadržavala 5,0 i hranu s oznakom 2 koja je sadržavala 10,0 mg fumonizina/kg. Kunice su takvu hranu dobivale tijekom dva tjedna prije parenja i četiri tjedna poslije parenja. Fumonizin u hrani značajno takvu hranu dobivale tijekom dva tjedna prije parenja i četiri tjedna poslije parenja. Fumonizin u hrani značajno je smanjio (P<0,05) dnevni unos suhe tvari i konačnu tjelesnu masu. Dnevni unos suhe tvari kunica hranjenih je smanjio (P<0,05) dnevni unos suhe tvari i konačnu tjelesnu masu. Dnevni unos suhe tvari kunica hranjenih hranom 1 bio je 6,32%, a onih hranjenih hranom 2 bio je 50,13% manji nego u onih kontrolne skupine. Broj hranom 1 bio je 6,32%, a onih hranjenih hranom 2 bio je 50,13% manji nego u onih kontrolne skupine. Broj eritrocita te vrijednosti hematokrita i hemoglobina bile su značajno smanjene (P<0,05), dok se broj leukocita eritrocita te vrijednosti hematokrita i hemoglobina bile su značajno smanjene (P<0,05), dok se broj leukocita u gravidnih kunica povećao poslije povećanja koncentracije fumonizina u hrani. Gravidne ženke hranjene u gravidnih kunica povećao poslije povećanja koncentracije fumonizina u hrani. Gravidne ženke hranjene kontrolnom hranom imale su značajno veću koncentraciju (P<0,05) ukupnih serumskih proteina nego one koje kontrolnom hranom imale su značajno veću koncentraciju (P<0,05) ukupnih serumskih proteina nego one koje su dobivale hranu 1 i 2. Aktivnost serumskih enzima značajno se (P<0,05) pojačala u kunica hranjenih hranom su dobivale hranu 1 i 2. Aktivnost serumskih enzima značajno se (P<0,05) pojačala u kunica hranjenih hranom 1 i 2. Hrana koja je sadržavala 5 mg fumonizina/kg smanjila je sposobnost rasta i uzrokovala negativne učinke 1 i 2. Hrana koja je sadržavala 5 mg fumonizina/kg smanjila je sposobnost rasta i uzrokovala negativne učinke na hematološke i biokemijske pokazatelje kunica što može poremetiti pravilan razvoj plodova.
na hematološke i biokemijske pokazatelje kunica što može poremetiti pravilan razvoj plodova.
Ključne riječi:
Ključne riječi: Fusarium verticillioidesFusarium verticillioides, fumonizin, haematologija, gravidne kunice, serum, biokemijski , fumonizin, haematologija, gravidne kunice, serum, biokemijski pokazatelji
pokazatelji
Received: 9 July 2009 Accepted: 15 March 2010
