Effects of Feeding Fusarium fujikuroi Culture Material, Containing Known Levels of Moniliformin, in Young Broiler Chicks1

Effects of Feeding Fusarium fujikuroi Culture Material, Containing Known Levels of Moniliformin, in Young Broiler Chicks1 D. R. LEDOUX,2-3 A. J. BERMUDEZ,4 G. E. ROTTINGHAUS, 4 and J. BROOMHEAD 2 Fusarium\Poultry

Research Laboratory, University of Missouri, Columbia, Missouri 65211

G. A. BENNETT

ABSTRACT An experiment was conducted with 270 male broiler chicks to evaluate the effects of a Fusarium fujikuroi M-1214 culture material containing moniliformin (M) on broiler chicks. Day-old chicks were allotted randomly to dietary treatments containing 0, .24, .48, .72, .96, 1.44,1.92, 2.40, and 2.88% M culture material (MCM). These levels of MCM supplied 0, 25, 50, 75,100,150, 200, 250, or 300 mg M / k g of feed. Each dietary treatment was fed to six pen replicates of five chicks per pen for 21 d. Significant mortality (P < .05) occurred in chicks fed 200 (8 out of 30), 250 (17 out of 30), and 300 (25 out of 30) m g M / k g feed. Chicks fed > 100 mg M / k g had lower (P < .05) feed intakes and smaller BW gains (P < .05) than controls. Increased heart weights (P < .05) were observed in chicks fed > 50 mg M / k g , and increased liver weights (P < .05) in chicks fed > 100 mg M / k g . Gross lesions of M toxicity included generalized cardiomegaly with dilation of the right ventricle. Histopathology revealed a high incidence of large and variably shaped cardiomyocyte nuclei and a generalized loss of cardiomyocyte cross striations in chicks fed > 75 and 200 mg M / k g , respectively. Results indicated that F. fujikuroi culture material containing M is toxic to young broiler chicks. (Key words: moniliformin, broiler, Fusarium fujikuroi, performance, cardiomegaly) 1995 Poultry Science 74:297-305 INTRODUCTION . , , .,. .. T i c mrM Received for publication July 5, 1994. Accepted for publication October 19,1994. Agricultural Experiment Station. Theofmention of firm ^ontribution Number 12144 the Missouri S ^ J ^ S S ^ S S f c ^ r f Agriculture over otherfirmsor similar products notmentioned. 3ZDepartment of Animal Sciences To whom correspondence should be addressed: 112 Animal Sciences Center, Columbia, MO 65211. 4 Veterinary Medical Diagnostic Laboratory. D

Fusarium moniliforme, a common contaminant of corn and other commodities, ' nas been linked to a number of disthe mycotoxins for these syn ease syndromesresponsible in poultry. However, dromes have not been identified. It has been speculated for some time that many of these unexplained syndromes may be caused by synergism between several . • » ; .•,•£ • /* *\ J c mycotoxins. Moniliformin (M) and fumonisin Bj (FBj) are two mycotoxins that are

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National Center for Agricultural Utilization Research USD A, Peoria, Illinois 61604

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5

Ken-Roy Hatchery, Berger, MO 63014. Fusarium Research Center, Pennsylvania State University, University Park, PA 16802. 7 Fitzmill Model D Comminuting machine, Fitzpatrick Co., Elmhurst, IL 60126.

and were characterized by myofiber degeneration and necrosis. Gross lesions included ascites, hydropericardium, and myocardial pallor (Engelhardt et al, 1989). The objectives of the present study were to investigate the toxic effects of F. fujikuroi cultures containing M in young broilers and to determine the minimum dietary concentrations necessary to cause deleterious effects. MATERIALS AND METHODS Experimental Design and Birds Two hundred and seventy day-old male Hy-Peco 5 broiler chicks were allotted randomly to pens in a stainless steel chick battery and allowed to consume feed and water ad libitum. The experimental design consisted of nine dietary treatments with six pen replicates of five chicks allotted randomly to each treatment. The day-old chicks were fed experimental diets from hatching to 21 d of age. Chicks were monitored daily for signs of morbidity and mortality. The animal care and use protocol was reviewed and approved by the University of Missouri-Columbia Animal Care and Use Committee. Moniliformin Production Cultures were grown in .946-L canning jars. Whole shelled corn (100 g) and 100 mL distilled water were added to each jar, autoclaved for 30 min at 121 C and a pressure of 1.4 kg/cm 2 , and allowed to cool to room temperature. Fusarium fujikuroi Nirenburg M-12146 was added to sterilized distilled water and 2 mL of the suspension was added to the autoclaved jars. The jars were shaken and lids loosened to allow for additional respiration. Cultures were then allowed to grow for 21 d at 25 C. At Day 21, jars were autoclaved and culture material dried at 40 C in a forced-air oven for 48 h, then ground to a fine powder in a mill 7 and stored at 0 C until used. Moniliformin Analysis

6

A ground sample (25 g) was extracted with 100 mL acetonitrile:water (80:20) for 60 min on a wrist action shaker. The solution

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known to be produced by F. tnoniliforme (Nelson, 1992). A recent report suggests that the combination of M and FBj may interact synergistically to produce what has been described as "spiking mortality" syndrome in broilers (Javed et al, 1993). Spiking mortality syndrome is characterized by a sudden onset of greater than 1% daily mortality for 3 to 5 d. Affected chicks develop opisthotonos, recumbency, depression, and hypoglycemia, and die within 2 to 6 h (Brown et al, 1991). Increased incidence of rickets and air sac disease is usually observed in affected flocks after mortality abates (Brown et al, 1991). Before the effects of multiple toxins can be evaluated, it is essential that each toxin be individually characterized and ruled out as being responsible for a syndrome. Research on the individual effects of FBj in chicks has shown that FBj was not by itself responsible for spiking mortality syndrome (Brown et al, 1992; Ledoux et al, 1992; Weibking et al, 1993a,c). In broilers, FBj is associated with poor performance, increased organ weights, multifocal hepatic necrosis, and biliary hyperplasia (Brown et al, 1992; Ledoux et al, 1992; Weibking et al, 1993b). With the exception of two reports (Allen et al, 1981; Engelhardt et al, 1989), initial studies of moniliformin toxicity in chicks involved determination of median lethal dose (LD50) values (Kriek et al, 1977; Burmeister et al, 1979). Allen et al. (1981) fed chicks pure M or M supplied by culture material at levels ranging from 8 to 64 m g / k g and observed reduced performance and mortality (15%) only in chicks fed 64 mg M / k g for 3 wk. Engelhardt et al (1989) fed chicks 0, 144, 288, or 576 mg M / k g supplied by culture material and reported 80% mortality by Day 10 in chicks fed 144 m g / k g , and 100% mortality by Day 10 in chicks fed 288 or 575 mg/kg. Lesions observed in chicks were most prominent in the ventricular myocardium

EFFECTS OF MONILIFORMIN IN BROILERS

Diet Preparation Dietary treatments were prepared by substituting F. fujikuroi M-1214 culture material for ground com in a typical cornsoybean basal diet. Moniliformin culture material (MCM) contained 10,500 mg M/kg by analysis and made up 0, .24, .48, .72, .96, 1.44,1.92, 2.40, and 2.88% of the respective diets and supplied 0,25,50,75,100,150,200, 250, and 300 mg M/kg diet, respectively. Diets were formulated to be isocaloric, isonitrogenous, and either met or exceeded the nutrient requirements of broiler chicks as recommended by the National Research Council (1984). Diets were screened and found to be free of aflatoxin, citrinin,

vomitoxin, fusarin C, sterigmatocystin, diacetoxyscirpenol, zearalenone, ochratoxin A, T-2 toxin, and the fumonisins. Sample Collection At the end of Weeks 1, 2, and 3 of the experiment, chicks were weighed individually and feed consumption was determined for each pen. On Day 21, blood samples were collected via cardiac puncture for serum biochemistry and hematologic determinations. Serum biochemical values were determined using an autoanalyzer.8 Hemoglobin was measured as cyanmethemoglobin.9 Red blood cell counts, mean corpuscular volume, and hematocrits were determined with a counter^ using instrument settings described by Steel et al. (1977). Mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) were calculated. Following blood sampling, 18 surviving chicks from each treatment were euthanatized by asphyxiation with C0 2 , and selected organs were excised and weighed. Histopathology Post-mortem examinations were performed on six surviving chicks from each treatment group at 3 wk of age. Chicks were anesthetized with CO2, euthanatized by cervical dislocation, and samples of lung, spleen, heart, kidney, and liver were fixed in 10% neutral-buffered formalin. Fixed tissues were trimmed, embedded in paraffin, sectioned at 4 jtm, and stained with hematoxylin and eosin stain. All tissues from the controls and from groups receiving MCM that supplied 200 and 250 mg M/ kg were examined microscopically. Liver, heart, and kidney sections from all experimental groups were examined microscopically, and heart and kidney lesions were graded in a blind fashion. Statistical Analysis

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Kodak Ektachem Analyzer, Eastman Kodak Co., Rochester, NY 14650. 'Sigma Diagnostics, Sigma Chemical Co., St. Louis, MO 63178-9916. 10 Coulter Counter Model ZB1, Coulter Electronics, Hialeah, FL 33010.

Data were analyzed by the General Linear Model procedure of SAS® software (SAS Institute, 1985) as a completely randomized design. Mean differences were determined using Fisher's least significant difference test. Absolute organ weights

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was filtered and a 10-mL aliquot transferred to a round bottom flask and rotoevaporated to near dryness. The residue was dissolved in 20 mL methanol and transferred to a separatory funnel. Hexane (25 mL) was added and the funnel was shaken for 2 to 3 min; phases were allowed to separate and the lower phase was returned to the round bottom flask and evaporated to near dryness. The residue was dissolved in 10 mL ion pair mobile phase (900 mL watenlOO mL acetonitrile:10 mL ion pair solution prepared by adding 25 mL tetrabutyl ammonium hydroxide to 50 mL 1.1 M dihydrogen phosphate buffer). A 1-mL aliquot of solution was filtered through a .2-fx syringe filter (13 mm) prior to analysis. Moniliformin analysis was performed on a Spectra Physics Model 8100 high-pressure liquid chromatograph equipped with a reversed phase C18 (5 /tm) column (4.6 x 250 mm), with detection by a diode array detector (230 nm). Flow rate of the mobile phase was 1 mL/min. A standard curve was prepared between 20 to 100 ng moniliformin. Sample extracts were diluted, as needed, so that absorbance values of moniliformin peaks (at 230 nm) fell within this range.

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were adjusted for final body weight by covariance analysis (Shirley, 1977). Statistical significance was accepted at P < .05. RESULTS

Compared with controls, significant mortality was observed in chicks fed MCM that supplied 200 (8 out of 30), 250 (17 out of 30), and 300 (25 out of 30) mg M/kg. As a result of the excessive mortality that occurred in chicks fed MCM that

supplied 250 and 300 mg M/kg, data from these treatment groups were not included in the statistical evaluation of response variables. The effects of dietary MCM on chick performance are presented in Table 1. Compared with controls, chicks fed MCM that supplied > 100 mg M/kg had reduced intakes and BW gains at Weeks 1, 2, and 3, and over the combined 3-wk period. Compared with controls, there was an increase in feed conversion in chicks fed MCM that supplied 100 and

Days

(""iiltiirp

Variable

\_UllUlC

1 to 7

8 to 14

15 to 21

1 to 21

0 (0)2 .24 (25) .48 (50) .72 (75) .96 (100) 1.44 (150) 1.92 (200) 2.40 (250) 2.88 (300) SE

116a 113a 110* 109a 99b 90 b 76c

270a 263" 282* 268a 220b 217b 186=

453 a 446a 455 a 438 a 391 b 376b 310=

839a 822a 847a 815a 712b 683 b 572=

3

9

8

16

0 (0) .24 (25) .48 (50) .72 (75) .96 (100) 1.44 (150) 1.92 (200) 2.40 (250) 2.88 (300) SE

106= 104* 98 bc 94c 83 d 75 e 64f

217a 213 a 227a 216a 174" 175b 141=

329a 330a 325 a 315a 287b 286 b 229c

651 a 647a 651 a 625 a 545 b 536" 436=

material (%)

Feed intake, g

3

Body weight gain- g

3

9

8

15

1.10= 1.09c l.llbc 1.17*c 1.18ab 1.20" 1.16*c

1.25 1.24 1.24 1.24 1.27 1.24 1.33

1.38 1.35 1.41 1.39 1.36 1.32 1.36

1.29 1.27 1.30 1.30 1.31 1.28 1.32

.03

.02

.03

.02

Feed conversion (feed:gain), g:g 0 (0) .24 (25) .48 (50) .72 (75) .96 (100) 1.44 (150) 1.92 (200) 2.40 (250) 2.38 (300) SE a_f

Values within columns with no common superscript differ significantly (P < .05). Data are means of six replicate pens of five chicks each less mortality. 2 Dietary levels (miligrams per kilogram) of moniliformin. 3 Due to excessive mortality, data were not used in statistical analysis. 1

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TABLE 1. Effects of Fusarium fujikuroi culture material, containing moniliformin, on performance of broiler chicks1

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EFFECTS OF MONILIFORMIN IN BROILERS TABLE 2. Effects of Fusarium fujikuroi culture material, containing moniliformin, on absolute organ weights of broiler chicks1-2 Culture material

Kidney

Spleen

5.17' 5.41de 5.80«i 6.07bc 6.54b 7.53* 7.92*

5.99 5.59 5.72 5.82 5.85 5.99 6.39

85* 78*bc 66c 80*b 70bc 84*

.17

.1!

.05

Liver

Heart

17.01 d 17.35d 17.65«< 18.34b«i 19.57b 18.85^ 21.12*

(g)-

(%) 0 (0)3 .24 (25) .48 (50) .72 (75) .96 (100) 1.44 (150) 1.92 (200) 2.40 (250) 2.88 (300) SE

6 9 bc

4

.52

a_e

150 mg M/kg at Week 1. Feed conversion was not affected by dietary treatment at Weeks 2 or 3, or over the combined 3-wk period. The effects of dietary MCM on-absolute organ weights are presented in Table 2. Compared with controls, increased liver weights were observed in chicks fed MCM

that supplied 100 to 200 mg M/kg, and increased heart weights were observed in chicks fed 50 to 200 mg M/kg. In contrast to liver and heart, effects of M on spleen weights were inconsistent, with M having no effect on spleen weights of chicks fed 50, 100, or 200 mg M/kg, whereas decreased spleen weights were observed

TABLE 3. Effects of fusarium fujikuroi culture material, containing moniliformin, on hematology of broiler chicks1-2 Culture material

RBC

HCT

HB

MCV

MCH

MCHC

(%)

(xl0«/mm3)

(%)

2.33 2.31 2.41 2.32 2.50 2.49 2.49 . .A

40.0 40.4 42.3 40.1 42.8 42.7 41.0

(lira?) 176* 175* 175* 176* 172* 172* 165b

(Pg)

0 (0)3 .24 (25) .48 (50) .72 (75) .96 (100) 1.44 (150) 1.92 (200) 2.40 (250) 2.88 (300) SE

(g/dL) 9.83* 9.69* 9.67* 9.10* 9.41* 8.73*b 7.91b

43.3* 42.3* 39.8*b 39.6*b 38.7*b 36.5bc 32.0'

(g/dL) 25.2* 24.3* 22.8*b 22.8*b 22.4*bc 21.2bc 19.4c

.08

1.3

.39

2

2.0

a_c

1.0

Values within columns with no common superscript differ significantly (P < .05). Data are means of six replicate pens of two chicks each. 2 RBC = red blood cells; HCT = hematocrit; HB = hemoglobin; MCV = mean cell volume; MCH = mean cell hemoglobin; MCHC = mean cell hemoglobin concentration. 3 Dietary levels (milligrams per kilogram) of moniliformin. 4 Due to excessive mortality, data were not used in statistical anlaysis. J

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Values within columns with no common superscript differ significantly (P < .05). Absolute organ weights adjusted for final body weight by analysis of covariance. 2 Data are means of six replicate pens of three chicks each. 3 Dietary levels (milligrams per kilogram) of moniliformin. 4 Due to excessive mortality, data were not used in statistical analysis. x

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Extramedullary hematopoiesis (EMH) was evident in the myocardium of chicks fed 50 to 300 mg M / k g , with the severity of this change ranging from mild focal

EMH to moderate multifocal EMH. Although isolated foci of EMH is a normal finding in the avian myocardium, the treatment-associated nature and severity of this change suggest that in this study EMH is a M-induced lesion. The right ventricular free wall of the heart was most severely affected by M toxicity, with cardiomyocytes arranged in harmonic wave motion pattern and a loss of cardiomyocyte cross striations (Figure 1). This harmonic wave motion of cardiomyocytes was most consistently evident in chicks fed 150 to 300 mg M / k g . A generalized loss of cardiomyocyte cross striations was also evident in chicks fed 200 to 300 mg M/kg. DISCUSSION In the present study, mortality was 13 and 83%, respectively, in chicks fed 150 and 300 mg M / k g . Significantly greater mortality was observed in previous studies with chicks fed M. Engelhardt et al. (1989) reported 80, 100, and 100% mortality by Day 9, in chicks fed MCM that supplied 144, 288, and 575 mg M / k g feed, respectively. Javed et al. (1993) reported 40 and 70% mortality, respectively, in chicks fed 27 and 154 mg pure M / k g . No mortality was observed in the present study in chicks fed 25 mg M / k g . Differences between the present study and that of Engelhardt et al. (1989) may be partially due to source of M or level of MCM used. Engelhardt et al. (1989) used MCM produced by F. moniliforme var. subglutinans at levels of 12.5 (144 mg M/kg) and 25% (288 mg M/kg) of the diet, whereas the source of M in the present study was F. fujikuroi at levels of 1.44 and 1.92% of the diet. The greater the level of culture material that is added to the diet the greater the chance that some other unknown mycotoxin or toxic factor is also being added to the diets. Higher mortality was observed by Javed et al. (1993) when they fed chicks levels of pure M comparable to those in the present study. It is possible that pure M has a greater bioavailability than the M present in culture material. In culture material a certain amount of M will be physically trapped in the center of parti-

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in chicks fed 25, 75, or 150 mg M / k g . Kidney weights were not affected by dietary MCM. Compared with controls, hemoglobin and mean corpuscular volume were decreased in chicks fed MCM that supplied 200 mg M / k g (Table 3). Chicks fed MCM that supplied 150 or 200 mg M / k g had lower MCH and MCHC than controls (Table 3). Red blood cell counts and hematocrits were not influenced by dietary MCM (Table 3). Serum concentrations of albumin, total protein, creatinine kinase, aspartate aminotransferase, uric acid, and sodium were not affected by dietary MCM (data not shown). Chicks fed rations containing 50 to 300 mg M / k g had a high incidence of generalized cardiomegaly. This lesion varied in severity from mild to moderate in each of these treatment groups, with isolated chicks having severe cardiomegaly. Dilation of the right ventricle was a prominent feature of chicks with moderate to severe cardiomegaly. One chick fed the ration containing 200 mg M / k g had ascites. The kidneys of three of five chicks necropsied in the group fed 300 mg M / k g in the ration were enlarged. Only five chicks were necropsied in this group because all the others in this treatment died prior to 21 d of age. Moniliformin-associated microscopic lesions were evident in the heart and kidney. Mild focal renal mineralization was evident in the kidneys of chicks fed 200 to 300 mg M/kg. Renal mineralization varied from mineralized casts in distal convoluted tubules to completely mineralized renal tubules. This lesion was generally mild and not interpreted to be of a sufficient severity to compromise renal function. Myocardial lesions were evident in chickens fed M at 50 to 300 mg/kg. The myocardial change that was the most sensitive indicator of M cardiac toxicity was cardiomyocyte nuclear morphology. Chicks fed 75 to 300 mg M / k g had a high incidence of variable-sized cardiomyocyte nuclei, with numerous large round and large oval nuclei (Figure 1).

EFFECTS OF MONILIFORMIN IN BROILERS

In contrast to the present study, in which BW gains were depressed at > 100 mg M/kg, Allen et al. (1981) reported a 14% decrease in BW gains of chicks fed MCM that supplied 64 mg M / k g and Javed et al. (1993) reported a 62% reduction in BW gains in chicks fed 27 m g / k g pure M. Lower M intake, as a result of lower feed intake, would be expected to have less of an effect on BW gains in the present study. The increase in heart weight observed in this study was observed in previous studies with chicks fed Fusarium proliferatum culture material that supplied 11 to 30 mg M / k g (Nagaraj et al, 1994) and turkeys fed culture material that supplied 25 to 330 mg M / k g (Ledoux et al., 1993; Reams et al, 1993). Gross pathology, increased heart weights, and histopathology are suggestive of generalized myo-

1 0

V

FIGURE 1. Photomicrographs (hematoxylin and eosin x 1,000) of heart sections of a control chick and a chick fed 300 mg moniliformin/kg feed, supplied by Fusarium fujikuroi culture material. Normal cardiomyocyte cross striations are evident in the control (A) and a loss of cardiomyocyte cross striations, harmonic wave motion of cardiomyocytes, and large nuclei (arrow) are evident in the chick fed moniliformin culture material (B). Bar equals 60 nm.

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cles of undigested culture material, and this M has no opportunity to be absorbed by the chick. Reductions in feed intake (15 to 31%) in this study were not observed in previous studies with chicks fed M supplied by MCM (Allen et al., 1981; Engelhardt et al., 1989). Reductions in feed intake observed in the present study may also partially explain the lower mortality because chicks with reduced intakes would have ingested less M. Reduced BW gains (16 to 36%) observed in the present study was less than that observed previously in chicks fed 144 to 575 mg M / k g (50%) supplied as MCM (Engelhardt et al, 1989) or chicks fed 27 mg (62%) or 154 mg (72%) pure M / k g (Javed et al, 1993). Greater reductions in BW gains were also observed in turkeys (50%) and ducks (75%) fed 144 to 575 mg M / k g (Engelhardt et al, 1989).

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cardial hypertrophy. Increases in liver toxicity. A similar loss of cross striations weights in response to MCM have also and cardiomyocyte wave pattern has been observed previously in turkeys fed previously been reported in turkeys with MCM that supplied 150 to 300 mg M / k g spontaneous cardiomyopathy (Gough et (Ledoux et al, 1993). al, 1981). Mild to moderate changes in hematoResults of this study are consistent with logic parameters were evident in chicks previous reports (Engelhardt et al, 1989; fed 150 to 200 mg M / k g (Table 3). The Cole et al, 1973; Nagaraj et al, 1994) most significant finding was a 19.5% indicating that M is cardiotoxic to poultry. decrease in blood hemoglobin concentra- The mechanism by which M causes cartion between controls and chicks fed diotoxicity has been investigated. Kriek et MCM containing 200 mg M/kg. This al (1977) noting clinical signs and the decrease in blood hemoglobin concentra- acute degenerative lesions in the myotion would significantly decrease the oxy- cardium of rats fed M suggested that M gen carrying capacity of the blood. A may suppress adenosine 5'-triphosphatedecreased blood oxygen carrying capacity d e p e n d e n t trans-membrane transport may result in the need for increased mechanisms, resulting in disturbances of cardiovascular output, which could ex- intracellular osmotic regulation and the acerbate the myocardial hypertrophy that consequent severe intracellular edema. In appears to be an intrinsic feature of MCM studies aimed at investigating this possitoxicity even at lower M levels. bility, Thiel (1978) found that exceedingly Rabie et al. (1982) suggested that in rats low concentrations of M (< 5 jtM) selecthere may be a threshold level of dietary tively inhibited rat liver mitochondrial M intake above which death is extremely pyruvate and a-ketoglutarate oxidations rapid. Extremely rapid death in chicks by 50% and suggested that these inhibiabove a threshold level of M (> 200 mg tory effects could constitute the major M/kg) suggests that such an acute toxicity molecular mechanism of toxic action. also occurs in chicks. The failure of MCM Based on the results of this study, it is to influence serum chemistry values may concluded that > 50 mg M / k g supplied by be associated with this phenomenon. The MCM is toxic to broiler chicks. It is also chronic effects of M toxicity were primar- concluded that M by itself is not responsiily manifested in chicks fed MCM that ble for spiking mortality syndrome supplied < 200 mg M / k g . This chronic (Brown et al, 1991), as the pathologic disease state may be less likely to produce changes of these two disease processes are the severe tissue damage of an acute markedly different. Specifically, the mytoxicity, and thus serum chemistry values ocardial changes observed in the present may be unchanged. study were not reported in spiking morThe pathology associated with M toxic- tality syndrome, and multifocal hepatic ity in other species has primarily involved necrosis, a prominent feature of spiking myocardial changes. In rats, the primary mortality syndrome, was not evident in lesion noted was acute myocardial de- the present study. generation and necrosis (Kriek et al, 1977). The current study indicates that levels Likewise, in young turkeys, chicks, and of M in excess of 50 m g / k g are toxic to the ducks, the primary lesion reported was young broiler chick, and that M by itself is acute myocardial degeneration and necro- not responsible for spiking mortality synsis (Engelhardt et al., 1989). This is in drome. The significance of this mycotoxin contrast to the cardiac lesions observed in to the poultry industry is difficult to assess this study, which were a loss of cardio- at this time because of the dearth of data myocyte cross striations, changes in car- on naturally occurring levels of M. In the diomyocyte nuclear morphology, ex- only two reports of naturally occurring t r a m e d u l l a r hematopoiesis, and cardio- levels to date, one researcher reported myocytes arranged in harmonic wave levels of 16 to 25 mg M / k g (Thiel et al, motion pattern in the right ventricular free 1982), whereas the second reported levels wall. The differences observed may be due of 17 to 425 mg M / k g (Logrieco et al, to the differences in chronic vs acute 1993).

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