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Molecular detection of quinolone resistance gene (gyrA) in Yersinia ruckeri isolates by PCR test
2016
Fadaeifard, Firooz | Nahid, Shahin | Momeni, Manochehr
BACKGROUND: Yersinia ruckeri is the etiological agent of enteric red mouth (ERM) or yersinioisis disease, one of the important bacterial diseases in the cultured salmonids. OBJECTIVES: The purpose of present study was detection of gyrA gene (quinolone resistance) in the Y. ruckeri bacterium. METHODS: In this study fish were evaluated in average size 8-12 cm from six rainbow trout farms in Chahar Mahal va Bakhtiyari province (Iran). In each farm 10 fish (totally 60) suspected to yersinioisis were randomly selected; sampling was done from lower part of intestine and cultured on Trpticase Soy Agar (TSA). The mediums were transferred to incubator and kept at 22 °C for 48 hours. Pure colonies which are grown on the mediums were tested by catalase, oxidase and gram staining, then those of gram-negative, catalase positive and oxidase negative were diagnosed, and cultured on Waltman- Shots medium (as specific medium for Y. ruckeri). These mediums were incubated at 22 °C for 48 h. Colonies that were grown were tested by PCR method for Y.ruckeri detection. Then, in the identified strains of Y.ruckeri gyrA gene were detected by PCR test. RESULTS: The results of bacteriological, biochemical and molecular tests showed that three cases out of total isolates were identified as Y. ruckeri. In all isolates of Y. ruckeri, gyrA gene was identified by molecular test. CONCLUSIONS: Identification of quinolone resistance gene in Y. ruckeri isolates can be the reason of low efficacy of these classes of antibiotics in the aquaculture. ِTherefore, the policy of treatment should be changed specially in enteric red mouth disease.
اظهر المزيد [+] اقل [-]Pathogenicity of Yersinia Ruckeri in Rainbow Trout (Oncorhynchus Mykiss)
2018
Taheri mirghaed, ali | Soltani, mehdi | Shafiei, Shafigh | Mirzargar, Seyed saeed | Shokrpur, Sara
BACKGROUND: Yersiniosis or enteric redmouth disease (ERM), caused by Yersinia ruckeri, is a serious bacterial disease in the farmed salmonids that causes economic problems in this industry. OBJECTIVES: This study was aimed to assess the experimental pathogenicity of Yersinia ruckeri in Rainbow trout (Oncorhynchus mykiss). METHODS: Two hundred Rainbow trout weighting 100-120 g, challenged with different strain of Yersinia ruckeri were obtained from affected trout farms using intra peritoneal injection route at a concentration of 108 cells/ml (0.1 mL per fish) to evaluate the virulence of these isolates. Each treatment group included 10 fish in two replicates and control fish received 0.1 mL sterile normal saline (0.9% NaCl). Following the intra peritoneal challenge, macroscopic and microscopic findings were determined. The most virulent strain was then used to determine the lethal concentration (LD50) using both intra peritoneal and bath method at dilutions of 103-1010 cells/mL. RESULTS: Macroscopically, anorexia, lethargy, circular swimming near the surface, blackening of skin, exophthalmia, hyperemia and hemorrhage in different parts of body, anal prolapse, enlarged liver and spleen were observed. Microscopically, hyperemia of hepatic sinusoids and vessels, necrosis and vacuolization of hepatocytes, increase in the abundance of macrophage centers in kidney, dilatation of Bowman’s space, degeneration and necrosis of kidney tubules, severe necrosis and detachment of intestinal villi, hyperplasia and clubbing of epithelial cells of secondary lamellae, spleen cell necrosis, goblet cell hyperplasia and thickening of epidermis layer in the tongue mucosa were observed. The LD50 of intra peritoneal injection was calculated 1.2×106cells per fish 48 h post challenge. In bath route, LD50 was obtained 5×108 Cells/ml after 96 h. CONCLUSIONS: The results obtained from this study show virulence diversity of native strains.
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