Toxicopathological Effects of Furazolidone on Ovaries of Adult Japanese Quail (Coturnix coturnix japonica)
Corresponding author: A. Khodakaram-Tafti, Department of Pathology, School of VeterinaryMedicine, Shiraz University, P.O. Box 71345-1731, Shiraz, Iran, Tel:0987132286950
Furazolidone (furoxone) is an antibacterial compound used for prevention and treatment of bacterial and proto- zoal diseases in medicine and veterinary medicine.In med- icine, it was used to treat cholera, giardiasis, diarrhea and gastro-enteritis conditions (Sweetman 2002, Hausen et al., 2006, Erlandsen and Meyer 2013). Trichomonasvaginalis causes vaginitis in woman and metronidazole-resistant iso- lates are sensitive to nitrofuranfurazolidone . Also, Giardia is susceptible to metronidazole and furazolidone but fura- zolidone was more effective by 50% than metronidazole in inhibiting in vitro cyst differentiation (Hausen et al.,2006).
Segura et al (1997) were reported that furazolidone therapy may be useful in Helicobacter pylori resistance to metroni- dazole and macrolide. Also, it is recommended furazolidone combination therapy for areas with high rates of resistance to Helicobacter pylori and for patients with multiple prior treatment failures (Zullo et al. 2012).
In veterinary medicine, despite prohibition of nitrofuranes administration in farm animals, these compounds synthe- sized and used as drug, food additive and antiseptic agent in some countries. The effect of treatment with different doses of furazolidone on the production, hatchability and fertility of eggs has been reported in chickens, turkey and Japanese quail(Ullah et al. 1998). Also, this drug cause to reduce weight and size of ovaries associated with small follicles in female Japanese quail (Ullah et al.1998). Also, var- ious degrees of degenerative changes and atrophy of testes associated with infertility were observed microscopically (Khodakaram-Tafti et al., 2015).
With regard to administration of furazolidone compounds for prevention and treatment of some bacterial and proto- zoal diseases in human and also some species of animals, this study was undertaken to elucidate role of these compounds in inducing toxicopathological lesions and infertility in mature female Japanese quail as a proper animal model.
Materials and Methods
Eighty adult (13 weeks old) female Japanese quails, were divided into 4 equal control and treatment groups. They were in good condition and clinically normal. Each group reared in similar environmental conditions. Furazolidone as a feed additive at 400, 800 and 1200 mg/kg to feed for a period of 14 days to treatment groups. One groupwas reared without fura- zolidone fed as control. Tenbirdsfromeach control and treat- ment groups afterone week and twoweeks were slaughtered. The ovaries of all birds were examined grossly. For histopatho- logical examination, the ovaries were fixed in 10% buffered formalin,embedded in paraffin, sectioned at 5 μm, stained with haematoxylin and eosin and studied microscopically. The degree of clinicopathological changes including reduction of egg production and size, reduction of follicles number in cor- tex, follicular atresia or degeneration, corpus atreticum or scar tissue was categorized as follows: (-) normal, (±) normal to mild, (+) mildly increase, (2+) moderately increase and (3+) severely increase (Table 1). The Chi-square test was used for comparison of the results and differences were considered as significant at P<0.05.
Grossly, in comparison to ovaries of control birds, the chang- es in groups received 800 and 1200 mg/kg during two weeks were including severe drop in egg production,various de- grees of ovarianatrophyassociated with prominent decrease in the number offollicles. No remarkable gross change was seen in 400mg/kg furazolidone-fed group.Histopathologically, ovaries of all fz-fed groups hadvarious degreesof atrophic small follicles with mild to most severely atresia. In comparison to control group (Figure 1),
Figure 1. Ovary; Adult Japanese Quail of control group; Normal cor- tical and medullary tissues with two round large developed follicles in the cortical tissue are observed (H&E stain, ×80).
the ovaries of group fz-400 had not remarkable microscopic lesions even after two weeks.
In this group, normal cortical and medullary tissues with a lot of small to large follicles were observed in the cortical tissue (Figure2 ).
Figure 2. Ovary; Adult Japanese Quail received 400 mg/kg furazoli- done after 2 weeks. The follicles are seen with normal architecuture and in various stages of development(H&E stain, ×80).
In fz-800 group after 1 week, the number of intermediate and developed follicles were decreased. Invasive atreticfollicles were observed prominently. After 2 weeks, the lesios were more remarkable including decrease in number of large devel- oped follicles, decrease in number of intermediate follicles so that many of them were wrinkled, atrophic or atretic (Figures 3& 4).
Figure 3. Ovary; Adult Japanese Quail received800mg/kg furazoli- done during 2 weeks. Remarkable reduce in the number of follicles, shrinkage of medium-sized follicles and increasing of invasive atretic follicles are seen (H&E stain, ×80).
Figure 4. Ovary; Adult Japanese Quail received 1200mg/kg furazoli- done during 2 weeks. Invasive atretic change in a developed follicle is seen (H&E stain, ×80).
The ovaries of group fz-1200 were most severely affected and had mostly atrophic oratretic follicles. Intermediate follicles were significantly reduced, wrinkled and were affected to in- vasive atretic changes (Figure 5). The number of large follicles were decreased prominently. The number of corpus atreticum (scar tissue) as fat storage tissue was increased instead of lost follicles (Figure 6) and some cases had accumulation of mela- nin like pigment in the medullary stromal tissue.
Figure 5. Ovary; Adult Japanese Quail received 1200mg/kg furazoli- done during 2 weeks.Intermediate follicles are wrinkled due to inva- sive atretic changes (H&E stain, ×80).
Figure 6. Ovary; Adult Japanese Quail received 1200 mg/kg furazoli- done during 2 weeks. Two corpus atreticum as fat storage tissues are seen instead of lost follicles associated with an atretic follicle (H&E stain, ×80).
The main results of this study after 2 weeksfurazolidonead- ministration are shown in table 1. By Chi-square test, no sig- nificant changes were seen in 400 mg/kg group, but signifi- cant changes were found in all criteria of 800 and 1200 mg/kg groups in comparison to control group (P<0.05).
Table 1. Main toxicopathological changes of furazolidon on ovaries of quail after 2 weeks.
|Groups||Dosage (mg/kg)||Number of adult
|Reduction of egg production and size||Decrease of follicles number||Increase of follicular atresia||Increase of corpus atreticum|
Abbreviations : (-) normal, (±) normal to mild, (+) mild, (2+) moder- ate and (3+) severe.
Furazolidone, N-(5- nitro- 2- furfurylidene)-3- amino -2- ox- azolidone, has been used for many years in the treatment of certain bacterial and protozoal infections in man and animals. The pharmacological, therapeutic and toxicological properties of furazolidone have been reviewed (Ali, 1999).
In human medicine, despite a diminution in its use in the re- cent years, the drug still prescribed for prevention and treat- ment of some infections. In the present study, we examined the effect of furazolidone on the ovaries of adult female japa- nese quail as a animal model.
Results of the present study showed furazolidone can causes various degrees of degeneration and atrophy of ovarian folli- cles associated with reducing the number of egg production and sizes of eggs even in therapeutic doses. Furazolidone caus- es free radical production, DNA irreversible injury, inhibition of cell replication and reduction of glutathione peroxidase rate in cell protection (De Angelis et al. 1999).
In chickens and turkeys, feeding different doses of furazolidone caused significant decrease in egg production, hatchability and fertility in a time and dose dependent manner (Ali,1999).
The therapeutic effect 400 and 600 mg/kg doses of furazoli- done during 4 weeks in female Japanese quail showed fura- zolidone feeding decreased body weight, feed intake and egg production. Grossly, the size and weight of ovaries, the height and number of mucosal folds of magnum, isthmus and uter- us had decreased compared with the control group (Ullah et al.,1998). The effects of feeding furazolidone on hematological and serum biochemical parameters of blood were reported in female adult Japanese quails (Nazifi and Asasi, 2001).
Dixon et al.. (1992) have reported significant reduction in egg production, egg quality, hatchability and fertility in breeding Japanese quail fed 200. 400 and 1000 mg/kg furazolidone for up to 33 days. The effects were being more pronounced at the higher doses. Reduction in hatchability has been both dose and time dependent and was attributable to an increase in infertile eggs rather than an increase in embryonic mortality. The high- est dose caused small-sized eggs to be produced.
Khan et al (2004) with morphometric studies on ovarian tissue of layer poultry birds (Gallus domesticus) revealed that the mean number of oocystes with diameter in the range 401- 800 µm decreased in birds recipient 400 to 800 mg/kg of fura- zolidone per day.
In the present study, different degrees of ovarian atrophy asso- ciated with degenerative changes and atresia of follicles were diagnosed at 800 mg/kg and 1200 mg/kg doses.
Kimaro et al (2013) investigated the effects of carbendazimon the magnum of female reproductive tract of Japanese quail and reported that it caused pyknosis and glandular atrophy in the mucosa. This drug had significant decline in the height of the mucosal folds, epithelial height, glandular width and glandular luminal diameter at 400 mg/kg and 800 mg/kg doses (P<0.05).
Ullah et al (1998), revealed that furazolidone toxicosis caused decreasing cell height in the mucosal glands in magnum and isthmus with centrally located nuclei and foamy cytoplasm. In the present study, with increasing dose of furazolidone,follicu- lar atresia particularly invasive type was increased in interme- diate and developed follicles.
In conclusion, furazolidone induced time and dose dependent, mild to severe toxicopathological effects on ovaries associated with remarkamle reduction in egg production and size. There- fore, with regard to toxic effects, reduction in fertility and pos- sible genotoxic and carcinogenetic effects of furazolidone and drug residual in meat, milk and egg even in topical forms, it is recommended that the administration of this drug in any form should be prohibited in veterinary medicine. Also, use of com- pounds contained furazolidone should be avoided in medicine.
The authors would like to sincerely thank the help of technical personnel of the Department of Pathology, School of Veteri- nary Medicine, Shiraz University,Iran.
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