And then returned to the tank with 120 liters of unpolluted water. Cohabitation challenge assaywas performedas follows: two rainbowtroutwere by intraperitoneal injection at a dose of 108 cfu, 1.6 fish1 infected as described above, when the first clinical published these two sick fish in a fig Kunststoffk placed under static inside the tank 120 L , containing the six healthy fish, there was no direct Lenalidomide TNF-alpha Receptor inhibitor contact between patients and healthy rainbow trout. The average time of 26 h cohabitationwas, began with the introduction of diseased fish and ended with the elimination of those who have died. Two groups of contr The, E and F were the same experimental procedures, subjected to the group B and C, except that sterile PBS was used instead of inocula.
In addition, w During the test period of not another group of six fish was manipulated and kept in the same physical conditions. Two groups of Nile tilapia, six fish were used in the test challenge. The members of the group I was by intraperitoneal AG-490 EGFR inhibitor injection with 0.2 ml of Weissella sp. Inoculum at a dose of 108 cfu fish1 1.2. The members of Group II were placed intraperitoneally with 0.2 ml of sterile PBS in question. A third group was kept under the same conditions as a control of experimental conditions. All challenge tests were performed in duplicate and the results are expressed as the mean for each repetition. The fish were washed four times t Monitors possible. The experimental period lasted 21 days, and all dead fish were subjected to bacteriological and histopathological evaluation.
At the end of the experiments, all surviving fish were by overdosing tert benzoca eingeschl And not subject to the same tests for microscopic L Look emissions and determine whether they asymptomatic carrier Were ger. All in vivo experiments Nelarabine were performed in accordance with the standards of animal welfare and were approved by the ethics committee in animal experimentation. 2.7. Bacteriological and pathological examination of samples of brain, kidney, liver, spleen, heart, eyes and intestines were taken away from her Aseptic we are all dead fish and surviving the end of the experiment. Those tissue ribbed fragment were 5% sheep blood agar and ontoMRS for bacterial reisolation. In addition, the bacterial load per gram of tissue in three organs was determined as follows: under sterile conditions, a small amount of each tissue was collected, weighed, k rperlich chtigten adversely, and in sterile PBS.
The tissue suspensions were serially diluted 10 times in sterile PBS, and 100 ml of each dilution were plated onto MRS. The plates were incubated at 25 8C for 48 h. The data was collected and in accordance cfu per gram of tissue the number of bacteria in serial dilution. The identification of bacterial isolates was again best of Weissella genus-specific PCR CONFIRMS. The pathological examination was performed for the same organs. The tissues were fixed in Bouin L Solution, embedded in paraffin wax and processed by routine methods. The sections were stained with H Matoxylin and eosin Rbt. were tested for 77 isolates with characteristic fragments of 798 bp. Phylogenetic analysis has been completed of the 16S rRNA gene sequence Shown born in the neighbor joining tree in F