The Fumonisins in Swine Production

Alberto Gimeno (albertogimeno@mail.telepac.pt)
Technical Consultant in Mycology and Feeding Mycotoxicology. Member of the Consulting Committee of Albéitar

The fumonisins

The fumonisins are toxic secondary metabolites, mainly produced by toxicogenic strains of Fusarium moniliforme. The greatest production of fumonisins take place in substrates with water activity, aw, greater than 0.91 and temperatures between 15° and 25° C. In general, Fusarium is a Genera of Fungi which belongs to the country flora (Phytopathogenic substrates of living plants) and the intermediate flora (humid fresh harvested cereal substrates).

There are six types of fumonisins: B1, B2, B3, B4, A1 and A2 (Marasas, 1995). The most frequent and important for their toxicity are the fumonisin B1 (FB1) and the fumonisin B2 (FB2). The latter ones could be found as natural contaminants in the cereal grains (mainly in corn and corn by-products). The fumonisins are resistant to high temperatures as high as 150° C, depending on their exposure time and pH of the substrate; they are very polar and soluble in water and acetonitrile.

The main syndromes produced (according to the specific animal specie) are: neurotoxic (leucoencephalomalacia), nephrotoxic, pulmonary and cerebral edema, hepatoxic and cardiac lesions. The affected organs are: brain, lungs, liver, kidneys and heart. These mycotoxins interfere with the sphingosine and sphinganine metabolism, causing a metabolic disturbance of the sphingolipids which are liver constituents and of lipoproteins (Prelusky et al, 1974; Marasas, 1995; Merrill et al, 1995; Merrill et al, 2001; Lino et al, 2004).

Occurrence of fumonisins in animal feeds

The FB1 is the most frequent natural contaminant, representing more than 70 % of all fumonisins found in cereal grains (mainly in corn) and their by-products. The FB2 is found in lower proportions as well as FB3.

Mallmann and Dilkin (2007), reported that in corn coming from the old Republic of Transkei (South Africa), found FB1 contaminant concentrations between 44 and 83 ppm (mg/Kg) and besides up to 117 ppm. In other studies performed in USA and South Africa the concentrations were 7.2 and 8.85 ppm, respectively and the presence of that mycotoxin concentrations in swine and horse feeds were found between 0.2 and 330 ppm.

Fifty percent of 64 feed samples in Uruguay were found contaminated with FB1 in concentrations ranging from 0.005 to 6.34 ppm. While in 50 samples of hybrid corn in Argentina, it was reported concentrations of the same mycotoxin of 100% of the samples between 0.185 and 27.05 ppm; and contaminations of FB2 between 0.04 and 9.95 ppm (Mallmann and Dilkin, 2007).

It seems that in Brazil, the greater amounts of fumonisins contamination in cereal grains and other feeds have been found in the Southern States; 50 to 90 % of the samples were positive and the contaminations ranged between 0.005 and 15 ppm, but the average was lower than 1 ppm and some contaminations of FB1 greater than 50 ppm had been found (Mallmann and Dilkin, 2007).

In the State of Rio Grande do Sul, also in Brazil, 267 samples of corn, 92 of mixed feed, 8 of oats, 5 of rice meal, 8 of soybean meal, 14 of barley and 13 of wheat were analyzed; the highest contamination of FB1 with the positive percentage for corn were 78.9 ppm and 35.2 %; for mixed feed 68.3 ppm and 30.4 %; for oats 0.175 ppm and 25 %; for rice meal 14.2 ppm and 75 %; for barley 2.4 ppm and 14.3 % and 2.4 ppm and 7.7 % for wheat. The samples for soybean meal were negative in the minimum detectable concentration according to the chromatographic analytical method used (0.05 ppm).

In Brazil, from a total of 11,814 samples of feedstuffs, the average fumonisins, adding both the FB1 and FB2, was 0.995 ppm. Corn samples represented 55.62 % of all samples and its average contamination was 1.01 ppm in 58% of this cereal. Mixed feed had an average of 1.426 ppm in 55% of the analyzed samples (Mallmann and Dilkin, 2007).

According to Mallmann and Dilkin (2007), the natural contamination in corn with FB1 and FB2, coming from several countries of Europe, were positive but low, showing contaminations between 0.055 and 5 ppm.

In USA, during 1995, the percentage and the FB1 contaminations in corn were 7 % with values above 5 ppm, 40 % between 0.5 and 5 ppm and 54 % with values lower than 5 ppm; the highest contamination was 24 ppm and the average of 1.5 ppm. In a survey carried out from 1996 to 1999 in south Georgia, up to 91% of the samples contained fumonisins; 30% of samples had concentrations above 1 ppm, and the highest contamination was 33 ppm. The World Health Organization working group found that globally 59% of corn and corn product samples were contaminated with FB1 (Hascheck et al, 2001).

In the approximately 106 mixed feed samples, with corn as a basic feedstuff, associated with confirmed cases of swine pulmonary edema in Brazil and the USA, had between 2 and 333 ppm of FB1 whereas FB2 levels in the five samples analyzed ranged from 1.9 to 28 ppm (Marasas, 1995).

In Hungary, almost 70 % of mouldy corn samples, since 1993, presented FB1 average contamination between 2.6 and 8.65 ppm and maximum concentrations between 9.8 and 75.1 ppm. It seems that the degree of contamination was increasing year by year (Zomborszky et al, 2000).

We should be alert, taking appropriate measures, after observing the information already given as well as the one that is going to be presented later in this article about the toxicity problems that these mycotoxins provoke in swine. Special attention should be taken mainly to control the mycotoxins which contaminate corn and corn by-products and consequently the mixed feed which include them, as the predominant cereal.

Fumonisins toxicity in swine

The fumonisins toxicity in swine is associated with pulmonary edema, hepatotoxicosis, nephrotoxicosis, cardiovascular and immunosupressors problems as well as alterations in the biosynthesis of sphingolipids and negative effects in daily feed consumption, body weight gain, conversion rate and carcass quality. Usually, the studies of the toxicity of fumonisins, most of the time, only the FB1 concentrations are given but we have also to take under consideration the frequent presence of FB2 along with FB1; the FB2 concentration is about from 15 to 35 % of the AFB1 (Hascheck et al, 2001). The presence of FB1, FB2 and FB3 has to be taken under consideration when we have to evaluate the problems associated with them.

Pulmonary edema syndrome

The contamination concentrations of mixed feed with FB1 could cause pulmonary edema problems in swine; in several occasions they are high but others that are lower with longer exposure time could also provokes those problems.

Mixed feed contaminations 92 ppm or greater cause pulmonary edema, after 4 to 7 days of feeding; with a calculated consumption of at least 6 mg of FB1 per Kg of body weight per day. About twelve hours prior to the development of the pulmonary edema and death of the animal, it was observed a state of inactivity, increment of the respiration rate and lowering of the cardiac rhythm (Haschek et al, 2001).

Some cases of pulmonary edema were observed in weaned pigs at lower contamination concentrations, between 10 and 40 ppm, but with longer feeding periods of 28 days (Hascheck et al, 2001).

Mallmann and Dilkin (2007), reported that levels of fumonisins were not detected in milk from sows infested with FB1 contaminated (greater than 100 ppm) feeds; the lactating pigs did not suffer from signs of intoxication. Other authors (Hascheck et al, 2001) point out that some pulmonary edema problems were observed in young pigs coming from two sows which were fed during a period of 14 days, from the 107th day of gestation to seven days after parturition, with contaminated feeds with 300 ppm of FB1.

Several symptoms of pulmonary edema were detected in a group of 20 weaned pigs which received contaminated feeds with FB1 in concentrations of 10, 20 and 40 ppm during a four week period; three animals had minor lesions at 10 ppm; two pigs had minor lesions and two pigs presented serious lesions in the group receiving 20 ppm; but five pigs showed serious lesions in the group which received the higher contamination of 40 ppm (Zomborszky et al, 2000).

In other experiments with weaned pigs, which received lower contaminations of 1, 5 and 10 ppm during 8 weeks resulted: at 1 ppm, one pig showed pathological alterations of the lungs, at 5 ppm, two pigs presented the pulmonary edema lesions and in the 10 ppm group, three pigs had the lung alterations; there were not significant clinical signs of importance and the pigs were not affected in their growth; however, the extended consumption of low FB1 contaminations produced lung alterations (Zomborszky et al, 2002).

Fumonisin B1 concentrations of 175 ppm (Becker et al, 1995) and 330 ppm (Fazekas et al, 1998) produced serious pulmonary edema cases even deaths. However is very difficult to find mixed feed with those high levels of contamination, at least in Spain and Portugal.

The inhibition of sphingolipid biosynthesis and the hepatic and cardiovascular problems

As we have above mentioned, the mechanism of toxic action of the fumonisins is the result of the interference with the sphingosine and sphinganine metabolism which result in a disturbance in the metabolism of sphingolipids. The sphingolipids have a great importance in the cell regulations and in the protein control at the cellular membrane level because they are present in them, they are the mediators of cellular growth and in their differentiation and death of cells. In mammals the concentration of sphingosine is, in general, about 3 to 5 times higher than sphinganine. The sphinganine N-aciltransferase and the sphingosine N-aciltransferase (ceramide syntetase) are fundamental enzymes in the biosynthesis metabolism of the sphingolipids (Lino et al, 2004).

The fumonisins could alter the concentration and the proportion between the sphinganine and the sphingosine; they can reduce the biosynthesis of sphingosines with the resulting accumulation of sphinganine. Those mycotoxins could block the biosynthesis of the sphingolipid complexes at the eucariotic cells. Those sphingolipid complexes are the base of the secondary messengers formation which control the different processes between cells, among those, the activation and deactivation of specific proteins and their genetic expression (Lino et al, 2004).

In swine, the partial or total inhibition of the sphingosine and the N-aciltransferase enzyme is responsible for the hepatoxicosis problems, so that the sphinganine/sphingosine ratio was indicated as a biomarker of the intoxication by fumonisins. Nowadays it is known that others mycotoxins can alter the ratio between these two sphingolipids (Mallmann and Dilkin, 2007).

The previous problems of pulmonary edema and provoked by high concentrations of FB1 in turn produces hepatic hydrothorax and icterus and the low contamination concentrations, but given at longer periods of time, produces liver necrosis and progressive degenerations which interfere in the proteic synthesis, body weight gain and the performance of the affected animals (Mallmann and Dilkin, 2007).

Swine fed with FB1 concentrations from 100 to 193 ppm during 93 days, developed a nodular hepatic hyperplasia, gastric ulcerations and cardiac and lung arteries hypertrophies. There were some cardiovascular variations when FB1 contaminations between 150 and 170 ppm were fed during a 217 day period, but they did not show hystopathological alterations in the heart. Swine fed with a mixed feed contaminated with fumonisin concentrations equal or greater to 23 ppm, suffered morphological alterations in the liver. (Haschek et al, 2001).

Other investigations by Mallmann and Dilkin (2007), reported that hepatic alterations like, aleatory hepatocellular necrosis, nuclear pleumorphism, mitosis increment, deformed cells, hepatomegalocitosis and focal hepatocitic necrosis are observed in higher frequencies in swine which suffer a chronic intoxication, with low fumonisin concentrations which are insufficient for the development of clinical pulmonary edema problems. The swine intoxicated with fumonisins have high concentrations of biochemical-clinic parameters such as, bilirrubines and the enzymes: aspartate aminotransferase, gamma-glutamil transferase, alkaline transferase, lactic and arginase dehydrogenases; all these reflect hepatoxicosis.

Some hepatic biochemical-clinic alterations were observed when FB1 and FB2 concentrations as low as 12 ppm were fed to pigs. Also, a hypercholesterolemia was reported when FB1 concentrations equal or greater to 1 ppm were given to other group of pigs. In other situations the hematologic parameters, like leucograms, hematocrit values and hemoglobin concentration were not significantly affected when pigs were intoxicated with fumonisins (Haschek et al, 2001).

The immunotoxicity of fumonisins

The presence of FB1 contaminations with concentrations of more than 20 ppm were associated with the occurrence of the PRRS (Porcine Reproductive and Respiratory Syndrome) disease in swine; there were 21 groups studied, twelve of them presented clinical symptoms of that disease and eight of them were consuming rations with more than 20 ppm of FB1; only one out of the nine that did not show the disease was consuming a diet with more than 20 ppm of FB1 (Mallmann and Dilkin, 2007).

The great susceptibility of swine to the infection by Pseudomona aeruginosa and the reduction of the concentration of macrophages in the lungs of young pigs fed for a week with mixed feed containing 20 ppm of FB1, were associated also with the immunotoxicity of this mycotoxin (Mallmann and Dilkin, 2007).

The fumonisins could provoke an increase in the susceptibility of the Escherichia coli; 35 Yorkshire cross young pigs, 3 weeks old and recently weaned which were fed during 6 days with a dose of 0.5 mg FB1/Kg body weight/day, which corresponds to a concentration of that mycotoxin of 5 to 8 ppm in the mixed feed; they presented a significant increase of the proliferation of the Escherichia coli, when it was administered orally. After 24 hours, that bacterial inoculation presented the greater colonization growth in the lungs, spleen, liver and kidneys and even greater in digestive organs like the jejunum, ileum, caecum and mesenteric lymphatic nodules; those animals did not present significant clinical symptoms, deaths nor important variations in body weight gains, when compared with the control group. The animals necropsies did not presented any macroscopic or microscopic lesions in the liver or in other tissues that could be related with the FB1 intoxications (Oswald et al, 2003).

Similar concentration contaminations of those mentioned above, but with longer contaminated periods, of about 56 days caused a decreased of 11% in daily body weight gains (Rotter et al, 1996).

Other undesirable effects of fumonisins

Barrows and gilts consumed FB1 contaminated mixed feed with 0.0; 0.1; 1.0 and 10 ppm, during 8 weeks. In general, the FB1 toxicity was more deleterious in barrows than in gilts. The barrows which consumed the rations with 1 and 10 ppm, presented lower body weight gains, 8 % and 11 %, respectively. The lowest FB1 contamination, 0.1 ppm, provoked in the barrows an abnormal growth during the first 5 weeks of the experiment; the feed consumption was somewhat higher than the control group during the first 4 weeks, but it was lower (6-7%) in each consecutive week.

The contaminations with 1 and 10 ppm, after 2 weeks, produced in the barrows an increase in the cholesterol; gilts receiving 1 ppm FB1 contaminated feeds, presented high cholesterol levels at the end of the 8 weeks. The barrows had heavier pancreas and suprarenal glands at the end of the 8 week period. An increment of the sphinganine and of the sphinganine/sphingosine ratio was observed. There were several problem cases of pulmonary edema in the higher FB1 contamination treatments (Rotter et al, 1996).

Several authors (Hascheck et al, 2001) point out that the variations in the carcass quality in swine could be negatively increased when they were consuming 1 ppm of FB1 contaminated feeds.

The effectiveness against fumonisins of a purified and modified phyllosilicate clay

Three trials were carry out in the Swine Area of the Universidade Federal de Santa Maria, Santa Maria, Brasil; Departamento de Medicina Veterinaria Preventiva, Laboratorio de Análises Micotoxicológicas-LAMIC, in order to test de efficacy of a purified and modified phyllosilicate clay against fumonisin in pigs.

In all trials, mixed feed and raw materials were initially analyzed for different mycotoxins: aflatoxins, ochratoxin A, zearalenone, deoxynivalenol, T-2 toxin and diacetoxyscirpenol. The results were negative in the minimum detectable concentrations according to the chromatographic analytical method used. Subsequently, mixed feeds not belonging to the control groups were experimentally contaminated with fumonisins. In one of the contaminated rations, a purified and modified phyllosilicate clay was added.

In the first trial, a total of 20 female piglets divided in several groups with 11.07 Kg initial body weight of 11.07 Kg/piglet, were fed 30 ppm of fumonisin FB1 contaminated feeds, during a 28 day period. There were not significant alterations in feed consumption, body weight gain and feed conversion or in the relative weights of liver or heart in the contaminated treatments when compared with the control group. The only significant alteration was in the relative weight of the lungs, going from 0.79g/100g (CV=6.89%) in the control group (non contaminated feed) to 1.22g/100g (CV=30.26%) in the contaminated group; in the group which received 30 ppm of contaminated feeds, containing also a 0.5% of a purified modified phyllosilicate, there was a relative lung weight of 0.87g/100g (CV=6.38%); this result suggests that the product significantly (P< 0.10) reduced the weight and the possibility of a pulmonary edema problem. No differences were observed between the control group and a group not contaminated but containing 0.5% of the phyllosilicate (LAMIC, 2006). As previously mentioned the FB1 toxicity is more deleterious in male pigs than females (Rotter et al, 1996)

In the second trial, a total of 12 male pigs divided in several groups with an average initial body weight of 58.33 Kg/pig, were fed 25 ppm contaminated feeds with fumonisins, during 28 day experimental period; at the end of the trial, when the contaminated treatment was compared with the control group, a reduction of the average daily feed consumption, average daily weight gain and the average body weight, 7.5, 26.7 and 9.1%, respectively, were observed; the conversion rate was also poorer in 20.3% and the relative average weights of the lungs, liver, spleen and heart increased 57.0, 13.7, 34.9 and 29.8%, respectively.

Other group male pigs which received also 25 ppm with fumonisins contaminated mixed feed was compared with a contaminated group which also received 0.4% of purified modified phyllosilicate; this latter group presented a significant (P< 0.05) improvement in their average feed consumption and feed conversion of 12.9 and 12.3%, respectively. That group, also presented average lower relative weights in lungs, liver and heart with 31.06, 20.7 and 22.9%, respectively; when compared with the control group, the phyllosilicate supplement group had only average relative weights in lungs and liver higher 8.26 and 1.57%, respectively and did show any difference with the average weight of the heart (LAMIC, 2008).

Another trial, using similar experimental details was performed, but extending the experimental period to 56 days; when the group receiving contaminated mixed feed was compared with the control group, a reduction was observed in average feed consumption, average body weight gain and average body weight of 5.7, 7.4 and 4.0%, respectively. The feed conversion was 7.1% poorer and the average relative weights of the lungs and heart increased 8.6 and 19.8%, respectively. When the blood samples were analyzed, the average plasmatic levels showed a reduction in Total Plasmatic Proteins (TPP) of 13%; the level of LDL cholesterol, increased 72.3% and the level of VLDL cholesterol decreased 47.2%; the sphinganine/sphingosine ratio increased 105.3%. The comparison between the contaminated groups, one with 25 ppm of fumonisins and the other with the addition of 0.4% of the purified modified phyllosilicate, the latter had significant (P< 0.05) increase in the average feed consumption, average daily weight gain and average body weight of 4.9, 8.8 and 5.3%, respectively. The feed conversion improved 7.2% and the average relative weight of lungs decreased 6.3%. The average plasmatic levels of the TPP increased 12.5% and the LDL cholesterol decreased 56.2%. The sphinganine/sphingosine ratio decreased 37.2%. When the phyllosilicate supplemented group was compared with control group, resulted in 1.04% reduction in the feed consumption and an increase in the average daily body weight gain and the body weight of 0.7 and 1.1%, respectively; the feed conversion improved a 0.6% and the average relative weight of the lungs increased only a 1.7%. The plasmatic levels, TPP and the LDL cholesterol, decreased 2.15 and 24.47%, respectively. The sphinganine/sphingosine ratio increased only 2.89% (LAMIC, 2008a).

Comments

Even though some high fumonisin concentrations (difficult to be find as natural contaminants) in short feeding periods of time provoke dangerous problems in swine, as pointed out previously, we have to take under consideration that substantial lower contaminations (easy and commonly found as natural contaminants) which are present over a longer feeding periods of time could cause the same problems. These situations make fumonisins, mainly the FB1, of great concern, as undesirable agents, in swine production.

Numerous studies are being carried with mycotoxin detoxifiers and should be further improved; the fact is that the majority of the efficacy studies have been evaluated "in vitro and not in vivo", mainly with respect of fumonisin mycotoxins. The results showed in this article regarding the purified and modified phyllosilicate clay, are "in vivo" and they demonstrated the effectiveness of the product against these mycotoxins in high concentrations of contamination.

In August 2006, a publication of the Official Journal of the European Union, recommended a guidance values for fumonisins B1+B2 in swine mixed feed (12% humidity) of 5 ppm, maximum.

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This article was published in ALBÉITAR (spanish veterinary journal), No. 116, June 2008, pp.54-57 and is going to be published in ALBÉITAR (portuguese veterinary journal), Vol.IV, No.4, July-August 2008