Leukoencephalomalacia in two horses induced by oral dosing of fumonisin B1

Leukoencephalomalacia (LEM) was induced by the oral administration of fumonisin B1 (FB1) to 2 horses: a filly received 59.5 mg/kg of a 50% preparation of FB1, administered in 21 doses of 1.25-4 mg/kg over 33 days; a colt, 44.3 mg/kg of 95% pure FB1 in 20 doses of 1-4 mg/kg in 29 days. Both animals developed nervous signs such as apathy, changes in temperament, inco-ordination, walking into objects, and one showed paralysis of the lips and tongue. Characteristic lesions of LEM were present in the brains. These trials proved conclusively that FB1 can induce LEM in horses.     

Pulmonary edema and hydrothorax in swine produced by fumonisin B1, a toxic metabolite of Fusarium moniliforme

Pulmonary edema and hydrothorax were observed in mature swine that died approximately 5 days after consuming corn screenings. These postmortem observations were reproduced in younger swine (16-24 kg) that died within 1 week when fed the corn screenings under experimental conditions. Additionally, pulmonary edema and hydrothorax occurred in a pig (7.1 kg) that died after receiving 4 daily intravenous injections of fumonisin B1. A fungus was isolated from the corn screenings that is identical to Fusarium moniliforme MRC-826 in colony morphology and under microscopic examination.     

Effects of Fusarium moniliforme culture material containing known levels of fumonisin B1 on progress of Salmonella Gallinarum infection in Japanese quail: clinical signs and hematologic studies

To study the individual and combined effects of fumonisin B1 (FB1) toxicity and Salmonella serotype Gallinarum infection, Japanese quail (Coturnix coturnix japonica) were fed Fusarium moniliforme culture material (2.5%), 150 mg FB1/kg ration, and were subsequently challenged orally with Salmonella Gallinarum organisms (2 x 10(4) colony-forming units) at 21 days of age. The chicks were fed culture material containing FB1 from day 5 till the end of the experiment. After being infected with Salmonella Gallinarum, observations were made 1, 2, 3, 5, 7, 10, 14, and 21 days postinfection. The clinical signs of diarrhea with bloody discharges were more pronounced in the Salmonella-infected birds on the FB1 diet. Mortality caused by Salmonella Gallinarum increased by 12% in the presence of FB1. Mean body weights in both the Salmonella-infected and FB1-fed groups were significantly lower than those of the controls at almost all intervals. Mean values of hemoglobin, packed cell volume, and total erythrocyte count were slightly higher in birds fed FB1 but were lower in the Salmonella Gallinarum groups fed FB1 and plain chick mash. Anemia was evident, between 5 and 10 days postinfection, in quail chicks infected with Salmonella Gallinarum alone. Total leukocyte counts were higher in Salmonella-infected and FB1-fed groups because of an increase in the number of heterophils and lymphocytes. However, the increase in lymphocyte response to infection was lower by 4.27%-30.09% between 3 and 21 days postinfection in the FB1-fed chicks compared with chicks infected with Salmonella Gallinarum. Alanine transaminase and total serum protein were slightly higher in both the infected and FB1-fed groups. This study revealed that the continuous presence of fumonisins in the diets of quail chicks might increase the susceptibility to or the severity of Salmonella Gallinarum infection.     

Individual and combined effects of Fusarium moniliforme culture material, containing known levels of fumonisin B1, and Salmonella gallinarum infection on liver of Japanese quail

Three hundred day-old Japanese quail (Coturnix coturnix japonica) were divided into two groups with 150 quail in each group. One group was maintained on quail mash alone, while Fusarium moniliforme culture material was added to quail mash in the second group from day 5 of age and was supplied at a rate of 150 ppm fumonisin B1 (FB1)/kg mash. At day 21, each group was further subdivided into two groups, yielding four groups with 75 birds apiece, which served as the control (group CX), the Salmonella Gallinarum alone group (group CS), the FB1 alone group (group FX), and the group fed FB1 and infected with Salmonella Gallinarum (group FS). An oral challenge with Salmonella Gallinarum organisms (2 x 10(4) colony-forming units/ml) was given to groups CS and FS at 21 days of age. Three quail each were necropsied on day 21 (0 day interval) from groups CX and FX only. At subsequent intervals (i.e., 1, 2, 3, 5, 7, 10, 14, and 21 days postinfection [DPI]), three quail were euthanatized from all four groups (CX, CS, FX, and FS). The gross and microscopic lesions were recorded in both mortality and euthanatized birds at the above intervals. The ultrastructural studies were done at 5 DPI. Mild to moderate hepatomegaly and pale discoloration of liver were observed in group FX, while congestion, hemorrhages, necrosis, and mild to severe hepatomegaly were the predominant gross lesions in both infected groups (CS and FS). The gross lesions in quail inoculated with Salmonella Gallinarum alone (group CS) generally developed slowly, appeared more widely scattered, and involved comparatively less surface area in contrast to the rapidly progressive and frequently confluent lesions in the combination group (FS), especially in the first 5 days of infection. Mild to marked hepatocellular swelling, multifocal hepatic necrosis, and hepatocellular and bile duct hyperplasia were the characteristic microscopic changes in the FX group. Microscopic lesions in quail of group CS comprised congestion, vacuolar changes, and focal necrosis in early stages, followed by granulomatous lesions at later intervals. Similar but more severe lesions were observed in the combination group (FS). Based on transmission electron microscopy, the maximum effect of FB1 toxicity was observed on mitochondria and endoplasmic reticulum. In general, the mitochondriae showed diverse form and structure, some of which appeared to lose their intact outer membrane, and the mitochondrial cristae were disoriented. The deformity in the cisternae structure of rough endoplasmic reticulum, with their rearrangement into round or tubular forms either bearing granular surface or leading to accumulation of smooth endoplasmic reticulum, was evident only in groups FX and FS. We conclude that the continuous presence of fumonisins in the diets of young quail might increase their susceptibility to or the severity of Salmonella Gallinarum infection.     

Individual and combined effects of the Fusarium mycotoxins fumonisin B1 and moniliformin in broiler chicks

The individual and combined effects of feeding fumonisin B1 (FB1; 0, 100, 200 mg FB1/kg) and moniliformin (M; 0, 100, 200 mg M/kg) were evaluated using a 3 x 3 factorial arrangement of treatments. Significant mortality (P < 0.05) occurred in chicks fed all diets containing 200 mg M/kg (50%-65%). Compared with controls and chicks fed FB1, both feed intake and body weight gain were decreased (P < 0.05) in chicks fed diets containing 100 mg M/kg. Chicks fed M had heavier heart weights (P < 0.05) than control chicks or chicks fed FB1. Compared with controls, chicks fed diets containing 200 mg M/kg or a combination of 200 mg FB1/kg and 100 mg M/kg had increased kidney and liver weights (P < 0.05). Significant FB1 by M interactions (P < 0.05) were observed for serum total protein and aspartate aminotransferase. Mild to moderate periportal extramedullary hematopoiesis and mild focal hepatic necrosis were observed in chicks fed FB1 alone. An increased incidence of large pleomorphic cardiomyocyte nuclei, loss of cardiomyocytes, and mild focal renal tubular mineralization were observed in chicks fed M alone. Both cardiac and renal lesions were observed in chicks fed combinations of FB1 and M. Data indicate FB1 and M, alone or in combination, can adversely affect chick performance and health at these dietary concentrations. The interactive effects of FB1 and M were not synergistic and were less than additive in nature. At the dietary concentrations studied, M is much more toxic to broilers than FB1.     

Microsporum gypseum isolated from ringworm lesions in a horse

Abstract

Extract

Microsporum gypseum has a world-wide distribution in soil, and it is considered to be essentially a soil inhabitant that only occasionally parasitizes animals and man (Georg, 1959 Georg, L. K. 1959. Animal Ringworm in Public Health, 3–17. U.S. Dept. of Health, Education and Welfare, Public Health Service Publ..  [Google Scholar]).     

Characterisation of Chlamydia psittaci isolated from a horse

This paper describes the isolation and characterisation of a strain of Chlamydia psittaci obtained from a nasal swab taken from a horse with serous nasal discharge. Initial isolation was achieved in cycloheximide-treated McCoy cell monolayers. Chlamydial inclusions stained by immunofluorescence either with a rabbit antiserum raised against C. psittaci or with a monoclonal antibody directed against the genus-specific lipopolysaccharide antigen were single and compact. They did not stain with iodine or with a monoclonal antibody reactive against Chlamydia trachomatis. The agent was re-isolated in the yolk sacs of embryonated hens eggs and designated N16. Identification of the agent was confirmed by electron microscopy. Unique plasmid DNA was prepared from a purified suspension of chlamydial elementary bodies (EBs), and analysed by electrophoresis through 1.0% agarose gels stained by ethidium bromide. This strain of C. psittaci grew relatively slowly in cycloheximide-treated McCoy cells, and the yield of elementary bodies during the course of one growth cycle was relatively low.     

Effects of prolonged exposure to low-dose fumonisin B1 in pigs

From the point of view of human exposure, fumonisins (FB1, FB2, FB1, FB4), a relatively recently (1988) discovered and identified group of mycotoxins, represent one of the five most important mycotoxin groups causing human disease. In an earlier experiment studying the effects of relatively low doses (10, 20 and 40 p.p.m.) of FB1 in weaned piglets, it was established that the 4-week feeding of 10 p.p.m. (mg/kg feed) FB1 produced mild pulmonary oedema. This suggested the importance of studies with even lower doses of the toxin to determine the tolerable limits. The objective of this experiment was therefore to study the effects of prolonged (8-week) exposure to still lower concentrations (0, 1, 5 and 10 mg/kg feed) of FB1. The 8-week feeding of FB1 in low concentrations (1-10 p.p.m.) did not cause clinical signs and significant performance impairment in pigs, but rendered irreversible the chronic changes that had already developed in the animals in a dose-dependent manner. Dissection revealed pathological alterations of the lungs in one of the animals given 1 p.p.m. (n = 4), in two animals exposed to 5 p.p.m. (n = 5), and in three animals given 10 p.p.m. (n = 4). In all three treatment groups, proliferation of the connective tissue fibres, primarily of those around the lymphatic vessels, in the subpleural and interlobular connective tissue of the lungs, extending to the peribronchial and peribronchiolar areas, was seen. The results of this experiment call attention to the risk of prolonged low-dose toxin exposure, which has very important public health implications.     

Intestinal barrier function is modulated by short-term exposure to fumonisin B1 in Ussing chambers

Veterinary Research Communications - Fumonisin B1 (FB1) alters intestinal epithelial cell cycle, absorptive, secretory and barrier properties of intestinal epithelial cells in vitro but data with...     

Effect of dietary fumonisin B1 on laying Japanese quail

1. A 28-d experiment was conducted to evaluate the effects of fumonisin B1 (FB1) on egg production and egg quality of young laying Japanese quail fed on fumonisin-contaminated rations. 2. To this end, 128 7-week-old birds were randomly distributed into 4 experimental groups (32 birds per group) and given rations containing 0 (control), 10, 50 and 250mg FB1/kg feed. Each treatment consisted of 4 replicates of 8 quail. Egg production and egg weight were checked daily. Feed consumption and feed conversion were determined weekly. Eggs laid on the last day of each 7-d period were collected and subjected to individual analysis for specific gravity, Haugh units and percentage eggshell. 3. Compared with controls, quail given > or = 50 mg FB1/kg had reduced feed intake and lower body weight gain. Feed conversion was reduced only in birds given 250 mg FB1/kg. 4. Mean egg production and egg weight were lower in birds given 250mg FB1/kg. Eggshell weight was reduced in birds given > or =50mg FB1/kg. However, mean specific gravity, Haugh units and percentage eggshell were not affected by FB1. 5. No histopathological changes were observed in liver, kidney or heart samples from any treatment group. 6. The results indicated that exposure to FB1 at concentrations > or = 50 mg/kg could adversely affect quail performance, emphasising the importance of controlling fumonisin contamination of quail rations.     

Cardiovascular changes associated with intravenous administration of fumonisin B1 in horses

OBJECTIVE: To determine whether cardiovascular dysfunction is evident in horses with leukoencephalomalacia experimentally induced by administration of fumonisin B1. ANIMALS: 11 healthy horses of various breeds (body weight, 252 to 367 kg). PROCEDURE: Horses were randomly assigned to 3 groups and administered fumonisin B1 daily. Horses received IV injections of 0 (control horses; n = 4), 0.01 (3), or 0.20 mg (4) of fumonisin B1/kg for 7 to 28 days. Horses were examined daily for evidence of neurologic disease. When neurologic signs consistent with leukoencephalomalacia were evident, horses were anesthetized, and catheters were inserted for evaluation of the cardiovascular system. After recovery from anesthesia, hemodynamic measurements were obtained. RESULTS: Fumonisin-treated horses with clinical signs of neurologic disease had evidence of cardiovascular dysfunction manifested as decreases in heart rate, cardiac output, right ventricular contractility (assessed by measuring the maximal rate of change of right ventricular pressure), coccygeal artery pulse pressure, and pH and base excess in venous blood as well as increases in systemic vascular resistance, compared with values for control horses. Fumonisin-treated horses with and without clinical signs of neurologic disease also had higher serum and right ventricular sphinganine and sphingosine concentrations than control horses. CONCLUSIONS AND CLINICAL RELEVANCE: An association was detected among fumonisin-induced neurologic disease, increased serum and myocardial sphinganine and sphingosine concentrations, and decreased cardiovascular function in horses. Fumonisin-induced decreases in cardiovascular function may contribute to the pathophysiologic development of leukoencephalomalacia in horses.     

Protective effects of silymarin on fumonisin B1-induced hepatotoxicity in mice

The present study was conducted to investigate the effect of silymarin on experimental liver toxication induced by Fumonisin B₁ (FB₁) in BALB/c mice. The mice were divided into six groups (n = 15). Group 1 served as the control. Group 2 was the silymarin control (100 mg/kg by gavage). Groups 3 and 4 were treated with FB₁ (Group 3, 1.5 mg/kg FB₁, intraperitoneally; and Group 4, 4.5 mg/kg FB₁). Group 5 received FB₁ (1.5 mg/kg) and silymarin (100 mg/kg), and Group 6 was given a higher dose of FB₁ (4.5 mg/kg FB₁) with silymarin (100 mg/kg). Silymarin treatment significantly decreased (p < 0.0001) the apoptotic rate. FB₁ administration significantly increased (p < 0.0001) proliferating cell nuclear antigen and Ki-67 expression. Furthermore, FB₁ elevated the levels of caspase-8 and tumor necrosis factor-alpha mediators while silymarin significantly reduced (p < 0.0001) the expression of these factors. Vascular endothelial growth factor (VEGF) and fibroblast growth factor-2 (FGF-2) expressions were significantly elevated in Group 4 (p < 0.0001). Silymarin administration alleviated increased VEGF and FGF-2 expression levels (p < 0.0001). In conclusion, silymarin ameliorated toxic liver damage caused by FB₁ in BALB/c mice.     

Hypotension in the horse induced by acepromazine maleate

The hypotensive effect of acepromazine maleate was related to both the dose and route of administration. The degree and duration of hypotension varied between individual horses. In general, intravenous administration produced a more rapid maximum effect than intramuscular injection and the larger the dose the longer blood pressure remained at low levels. Average systolic pressure (of 6 horses) remained significantly below control values for more than 6 h after an intramuscular injection of 0.05 mg acepromazine maleate/kg body weight. Respiratory rate was markedly decreased, with the duration, but not the degree of bradypnoea related to the dose administered. A slight, transient decrease in body temperature was observed. Heart rate was not significantly influenced by administration of acepromazine maleate.     

Neurologic abnormalities and cerebrospinal fluid changes in horses administered fumonisin B1 intravenously

The objective of this experiment was to characterize a dose-dependent toxic effect of fumonisin B1 (FB1) and to document initial neurologic signs, clinical progression, and terminal cerebrospinal fluid (CSF) changes in horses administered FB1 IV. Seventeen healthy horses were administered 0.00 (n = 4), 0.01 (n = 3), 0.05 (n = 3), 0.10 (n = 3), or 0.20 mg (n = 4) of purified FB1 IV q24h. When neurologic abnormalities observed by a masked observer became severe, atlanto-occipital CSF taps were performed and CSF pressure, cell count, cytology, protein, albumin and glucose concentrations, and creatine kinase activity were measured. Changes in CSF and number of days to 1st observation of neurologic abnormalities were compared between doses by ANOVA, with the level of significance set at P < .05. Control horses and low-dose horses (0.01 mg/kg) remained neurologically normal. In higher dose FB1-treated horses (n = 10), initial clinical signs (days 4-10) included hindlimb ataxia, delayed forelimb placing, and decreased tongue tone and movement. Hindlimb and trunkal ataxia, depression, hyperesthesia, and intermittent dementia gradually became apparent. When data from all horses with neurologic abnormalities were pooled (0.05-0.20 mg/kg FB1), mild clinical signs (mean day 6.3) occurred significantly earlier than did more severe (mean day 8.9) clinical signs (P = .009). Neurologic horses had high CSF protein, albumin, and IgG concentrations and increased albumin quotients (P < .05). It was concluded that FB1-induced neurologic and CSF changes in a dose-dependent manner, with a no-observable-limit of 0.01 mg FB1/kg IV q24h for 28 days. The neurologic and CSF changes were consistent with vasogenic cerebral edema.     

Growth performance and puberty attainment in growing pigs fed dietary fumonisin B1

Twenty‐four male weaned piglets assigned to four diets containing 5.0, 10.0, 15.0 and 0.2 mg fumonisin B₁ (FB₁) /kg as diets 1, 2, 3 and control diet, respectively, were used to study the effect of dietary FB₁ on growth and puberty attainment in pigs in a 6‐month feeding trial. Lower feed intake during 0–4 months and a non‐significant (p > 0.05) but FB₁ concentration‐dependent decrease in live and DWGs in animals fed FB₁‐contaminated diets was observed at the end of the pubertal phase. The daily and the final live weight gains of animals fed diet 3 were 75.8% and 90.6%, respectively, of the control values. The mean ages at puberty by boars on diets 2 and 3 were significantly (p < 0.05) higher than those for animals on the control and diet 1. The animals on diet 3 attained puberty when mean live weight was 60.1 kg, some 30.3 days after the controls attained puberty, at 156.3 days, when the mean live weight was 46.9 kg. This study revealed that dietary FB₁ delays sexual maturity in growing pigs. Male weanling pigs for breeding should not be exposed to dietary FB₁ higher than 5 mg/kg for optimum growth and reproductive performance.     

Experiment to determine limits of tolerance for fumonisin B1 in weaned piglets

In Hungary almost 70% of mould-affected maize inspected since 1993 was found to be contaminated with fumonisin B1 (FB1) (mean 2.6-8.65 mg/kg; maximum 9.8-75.1 mg/kg), the degree of this contamination was found to increase from year to year (Fazekas et al., 1997b). In this experiment, in order to define tolerance limit values, the effect of exposing weaned piglets to FB1 in low doses over a 4-week period was examined. The experiment was performed with 20 weaned barrows of Danish Landrace breed. After a 5-day adaptation period cultures of the fungus Fusarium moniliforme were mixed into the animals' feed in concentrations that resulted in a daily intake of fumonisin B1 of 0, 10, 20 and 40 mg/kg feed. Feeding with the toxin was observed to exert no significant effect on body weight gain or feed consumption in the animals, no clinical signs were observed and no mortality traceable to toxic effects occurred. In computer tomography examinations performed in the second and fourth weeks mild and more severe pulmonary oedema was diagnosed in the experimental animals. The processes developing in the pulmonary parenchyma were corroborated by the mathematical and statistical evaluation procedures applied. The haematological parameters examined revealed no change attributable to toxic effects, while with respect to the biochemical parameters, an increase in aspartate aminotransferase (AST) activity dependent on dosage, indicating a pathological change in the liver, was ascertained in all three experimental groups. The free sphinganine to sphingosine ratio (SA/SO), which is regarded as the most sensitive bioindicator of fumonisin toxicosis, showed an increase proportionate to toxin concentration for all three dosages. Dissection revealed mild cases of pulmonary oedema in three of the animals given doses of 10 p.p.m. (n = 4), two mild and two severe cases in those exposed to 20 p.p.m. (n = 5), and severe cases in all five animals given 40 p.p.m. The oedema of non-inflammatory origin was confirmed by histopathological examinations. The findings of this experiment which indicate that in this study FB1 administered in substantially lower concentrations than those reported in the literature resulted in severe pathological changes, point to the importance of studies involving even lower doses.