1 induced nephropathy in Japanese quail 1

Aim: The present study was designed to evaluate the toxic interaction of fumonisin B and ochratoxin A, alone and in 1 combination for pathology of the kidneys in day old Japanese quail. Materials and Methods: Day old Japanese quail chicks were divided into 4 groups and fed with basal quail chick mash containing fumonisin B @ 200 ppm (Group FX), Ochratoxin A @ 2 ppm (Group OX), and fumonisin B @ 200 ppm + 1 1 Ochratoxin A @ 2 ppm (Group FO) and standard toxin free feed (Group CX, control) for 28 days. Results: The quail chicks were assessed for various microscopic changes in the kidneys. Mean microscopic score values for congestion was found to be significantly higher (P< 0.05) in FO group when compared to that of other groups FX and OX at 7 and 21 DPF. Similarly, mean microscopic score values for degeneration was found to be significantly higher (P < 0.05) in FO group as compared to that of groups FX and OX at 7 and 14 DPF. Mean microscopic score values for necrosis and luminal hyaline bodies were significantly higher (P < 0.05) in FO group when compared to that of groups OX and FX at 7 DPF. Conclusion: Total score card for microscopic lesions observed in the kidneys when compared between different treatment groups showed that the lesions appeared to be more pronounced in the OTA fed group at 14 DPF as compared to other treatment groups. The overall lesion score values after the conclusion of the experiment were found to be higher in the combination group followed by the groups OX and FX, respectively.


Introduction
with respect to the changes induced by them in lymphoid and other organs of poultry [5,6] limited The fumonisins were isolated for the first time information is available on the pathologic effects of from the fungus Fusarium moniliforme (now renamed fumonisins on organs other than the liver.Fusarium verticillioides) in South Africa.A number of Ochratoxins, produced mainly by Aspergillus fumonisins have since been isolated and ochraceus (now called A. alutaceus) and Penicillum characterized, but FB remains the most toxic 1 verrucosum, are chemically characterized as 3, 4 compound [1].FB either in naturally contaminated 1 dihydromethylisocoumarin derivatives linked with an maize or maize-based feeds or in purified form, has amide bond to the amino group of L-D phenylalanine.been reported to cause equine leukoencephalomalacia Ochratoxin-A (OTA) is more toxic than other [2], porcine pulmonary edema and hydrothorax ochratoxins (B and C) and has been reported to be syndrome [3].The majority of reports on the toxic more toxic than even aflatoxin B [7].An OTA causes 1 effects of FB in avian species pertain to chickens, 1 significant loss to the poultry industry due to its effects ducklings and turkeys, in which primary pathological on performance and health.It causes a reduction in changes have been reported in the liver characterized growth rate and feed consumption, poorer feed by multifocal hepatic necrosis and hepatocellular and conversion and increased mortality [8].OTA induces biliary hyperplasia [4].Unlike aflatoxins and degenerative changes and an increase in weight of the ochratoxins, which have been studied extensively kidneys and liver, as well as decrease in the weight of NRRL-3174 (Courtesy: Dr. G. E. Rottinghaus, The effects of FB and OTA in combination have University of Missouri, Columbia, USA).Fusarium 1 culture material containing 6200 mg FB per kg and not been investigated in Japanese quail.The purpose 1 of this study was to investigate the toxicity and ochratoxin culture material containing 80 mg OTA per describe the major effects of feeding quail chicks diets kg was incorporated at the rate of 3.25 per cent and 2.5 containing FB and ochratoxin A alone and in per cent in the chick mash to supply 200 ppm FB and 2 were kept under strict hygienic conditions throughout 75 birds in each of the above mentioned four groups.the period of the experiment.The animal care and The present study was conducted using three pen experimental protocol were approved by the replicates of 25 quail per pen in each of the four groups University and by the Committee for the Purpose of for a period of 28 days.Various dietary treatments Control and Supervision of Experiments on Animals starting from day one until the end of the experiment (CPCSEA).
are presented in the tabulated form in Table -1.Feeding schedule: The quail chicks were maintained Total and mean lesion scores: Total microscopic on chick mash (Quail mash procured from the lesion scores for the sacrificed birds were determined Department of Animal Nutrition, COVAS, CSK at various time intervals by sum total of all the lesion HPKV, Palampur) from day one until the end of the scores obtained by multiplying the intensity score by experiment.Feed was autoclaved for 15 minutes at 15 the number of birds showing that particular intensity pounds pressure before feeding or mixing with of lesion.The lesion score for each of the microscopic Fusarium culture material (s).Boiled (for 15 minutes) lesions in different groups were subjected to statistical and subsequently cooled water was given to the birds test for calculation and comparison of mean lesion throughout the experiment.Feed and water were given scores among different groups at 95% confidence ad libitum, and no medication was given during the level (P < 0.05).period of the experiment.Before feeding, the representative samples of chick mash were submitted Statistical analysis of data: Data were subjected to to the Animal Feed Analytical and Quality Control statistical analysis for drawing inferences using a Laboratory, Veterinary Hospital Campus, Trichy standard procedure [10].Treatment means were Road, Namakkal, Tamil Nadu, India for the analysis of compared using Duncan's Multiple Range Test (2-way common mycotoxins, and to the Department of ANOVA) to determine the effect of treatment, age and Veterinary Preventive Medicine and Epidemiology, their interactions [11].All levels of significance were CCS HAU, Hisar for analysis of fumonisins contents.based on the 95 per cent level of probability.The feed samples were found to contain 12 ppb of Results aflatoxin-B and were free from ochratoxin-A, Microscopic pathology: When total score card for aflatoxin-G and T-2 toxin.The mycotoxins for the 2 microscopic lesions observed in the kidneys was present studies i.e. fumonisin-B and ochratoxin-A compared between different treatment groups, the 1 were supplied by Fusarium verticillioides M-1325 lesions appeared to be more pronounced in the OTA culture material (FCM) and Aspergillus ochraceous fed group at 14 DPF as compared to other groups.The overall lesion score values after the conclusion of the the end of the experiment revealed significantly higher experiment were found to be higher in the combination (P < 0.05) congestion of kidneys in group FO when group followed by groups OX and FX, respectively compared with the other treatment groups FX and OX.(Table -2).The overall treatment effect in relation to progressing When mean microscopic score values for age of birds on the occurrence of congestion in the congestion observed microscopically were compared renal interstitial tissue was found to be highly between different treatment groups, the difference was significant (P < 0.01) (Table-3).found to be significantly higher (P < 0.05) in FO group When mean microscopic score values for degeneration were compared between different when compared to that of other groups FX and OX at 7 treatment groups, the difference was found to be and 21 DPF ( significantly higher (P<0.05) in FO group as occurrence of necrosis in the kidneys was found to be compared to that of groups FX and OX at 7 and 14 DPF highly significant (P <0.01) (Table -

5). (Table-4). At 21 DPF, the comparative values in the
Mean score values for luminal hyaline bodies group FO were significantly higher than any of the when compared between different treatment groups, groups fed either FB or OTA alone.The mean the difference was found to be significantly higher (P < 1 0.05) in FO group when compared to those of groups treatment effect at the end of the experiment revealed OX and FX at 7 DPF (Table 6).The mean treatment significantly higher (P<0.05)degeneration in the effect at the end of the experiment revealed kidneys in group FO when compared to those of significantly higher (P<0.05)occurrence of groups FX and OX.
eosinophilic bodies in the renal tubules in group FO When the mean microscopic score values for when compared to those of groups FX and OX.necrosis were compared between different treatment groups, the difference was found to be significantly Discussion higher (P <0.05) in FO group when compared to that of The microscopic lesions noticed in the present groups OX and FX at 7 DPF (Table-5).The mean treatment effect at the end of the experiment revealed study in the FB fed group were more or less in 1 significantly higher (P<0.05)values for the conformity with the earlier reports [12] who reported occurrence of necrosis in group FO when compared to mild focal renal tubular mineralization in broiler those of groups OX and FX.The overall treatment chicks fed 100 and 200 ppm FB for 21 days.Similarly 1 effect in relation to progressing age of birds on the [13]  changes in tubular epithelial cells and occasional additive in nature for most of the microscopic infiltration with heterophils in the interstitial tissue variables except for interstitial congestion and near degenerating tubules in quail fed FCM supplying degenerative changes which yielded an additive 150 mg FB /kg mash for 21 days and demonstrated interaction at 21 DPF.
1 that FB inhibits enzyme ceramide synthase in the 1 Author's contribution kidneys [14] with a consequent increase in SA MAK and RKA implemented the study design.MAK concentration and SA/SO ratio.The mechanism of and AI done the Statistical analysis of data and renal toxicity induced by FB has been reported to be 1 collection of research material.RKA had given the quite different from that of OTA.Altered sphingolipid guidance during the study.MAK, AI and RKA drafted metabolism affecting the cell membrane integrity and the manuscript and revised the manuscript.All author functions of the epithelial cells was reported to be the read and approved the final version of manuscript.changes preceding even before the development of the lesions evidenced histopathologically.
1 citrinin, zearalenone, aflatoxin-B , aflatoxin-G , 2 1 Three hundred, one-day-old conducted on three hundred one-day-old Japanese Japanese quail chicks were randomly divided into four quail chicks procured from the Central Poultry groups i.e.FX (fumonisin B ), OX (ochratoxin-A), FO Materials and Methodsincorporated in the diet of control group (CX).Experimental birds: The present studies wereExperimental design:

Table - 1
. Table representing dietary treatments starting from day one until the end of the experiment.

Table - 2
. The mean treatment effect at .Total microscopic lesion scores in the kidneys of Japanese quail at different intervals FX = fumonisin B only, OX = ochratoxin A only, FO = fumonisin B + ochratoxin A, DPF = days post-feeding

Table - 3
. Interstitial congestion indicated by mean microscopic lesion score in different groups of Japanese quail Values within a column with different superscripts showing mean treatment effect are significantly different by ANOVA (P < 0.05) HSF-value indicating interaction between different treatments and age of quail chicks (HS = highly significant) by ANOVA (P <0.01).A-BValues within a row with different superscripts showing mean age effect are significantly different by ANOVA (P < 0.05).1.Data are means ± SE of three replicate pens of 3 quail each.CX = birds fed quail mash alone; FX = birds fed fumonisin B1; OX = birds fed ochratoxin A; and FO = birds fed fumonisin B1 and ochratoxin A. Values within a row with different superscripts showing mean age effect are significantly different by ANOVA (P < 0.05).1.Data are means ± SE of three replicate pens of 3 quail each.CX = birds fed quail mash alone; FX = birds fed fumonisin B1; OX = birds fed ochratoxin A; and FO = birds fed fumonisin B1 and ochratoxin A.
a-c: Values within columns (between groups CX, FX, OX and FO) with different superscripts are significantly different by ANOVA (P 0.05).x-z: Table-4.Degenerative changes in the kidneys indicated by mean microscopic lesion score in different groups of Japanese quail a-d: Values within columns (between groups CX, FX, OX and FO) with different superscripts are significantly different by ANOVA (P < 0.05).x-z: Values within a column with different superscripts showing mean treatment effect are significantly different by ANOVA (P < 0.05) HSF-value indicating interaction between different treatments and age of quail chicks (HS = highly significant) by ANOVA (P < 0.01).A-B:

Table - 6
revealed progressive degenerative and vacuolar Renal tubular epithelial cell necrosis indicated by mean microscopic lesion score in different groups of Values within columns (between groups CX, FX, OX and FO) with different superscripts are significantly different by ANOVA (P < 0.05).x-z: Values within a column with different superscripts showing mean treatment effect are significantly different by ANOVA (P < 0.05) HSF-value indicating interaction between different treatments and age of quail chicks (HS = highly significant) by ANOVA (P < 0.01).A-DValues within a row with different superscripts showing mean age effect are significantly different by ANOVA (P < 0.05).1.Data are means ± SE of three replicate pens of 3 quail each.CX = birds fed quail mash alone; FX = birds fed fumonisin B1; OX = birds fed ochratoxin A; and FO = birds fed fumonisin B1 and ochratoxin A. .Occurrence of eosinophilic hyaline bodies in the lumen of renal tubules indicated by mean lesion score in different groups of Japanese quail Data are means ± SE of three replicate pens of 3 quail each.CX = birds fed quail mash alone; FX = birds fed fumonisin B1; OX = birds fed ochratoxin A; and FO = birds fed fumonisin B1 and ochratoxin A.
a Table-5.a-d: Values within columns (between groups CX, FX, OX and FO) with different superscripts are significantly different by ANOVA (P < 0.05).x-z: Values within a column with different superscripts showing mean treatment effect are significantly different by ANOVA (P < 0.05) HSF-value indicating interaction between different treatments and age of quail chicks (HS = highly significant) by ANOVA (P < 0.01).A-B: Values within a row with different superscripts showing mean age effect are significantly different by ANOVA (P < 0.05).1.