Enrofloxacin, Effective Treatment of Pseudomonas aeruginosa and Enterococcus faecalis Infection in Oreochromis niloticus.

Enrofloxacin is a broad-spectrum synthetic antimicrobial drug widely used in veterinary medicine. The present study aimed to determine the effective enrofloxacin dose for treating Pseudomonas aeruginosa and Enterococcus faecalis infection in Oreochromis niloticus. P. aeruginosa and E. faecalis isolates were verified using selective differential media and biochemically using the Vitek 2 test. Bacterial isolates were virulent for O. niloticus with LD50 equal to 2.03 × 106 and 2.22 × 107 CFU fish-1 for P. aeruginosa and E. faecalis, respectively. Infected fish suffered from decreased feed intake followed by off-food, tail erosion, darkening of the external body surface, exophthalmia, ascites, and loss of escape reflex. Internally, congested hemorrhagic hepatopancreas with engorged distended gall bladder were dominant. The posterior kidney was congested with enlarged spleen, and empty elementary tract. Pathologically, severe degenerative changes were dominant in the hepatopancreas, posterior kidney, spleen, stomach, and gills of infected fish. Antimicrobial sensitivity test indicated the high susceptibility of P. aeruginosa and E. faecalis to enrofloxacin with MIC estimated at 1 and 0.0625 µg/mL, respectively. Enrofloxacin effectively protected O. niloticus against E. faecalis and P. aeruginosa infection when used with medicated feed at doses of 10 and 20 mg kg-1 body weight.

The role of Aeromonas genotyping in virulence for Dicentrarchus labrax.

Aeromonas septicemia still represents a serious challenge facing the global aquaculture sector. In the present study, Aeromonas caviae and A. veronii were isolated from four diseased European seabass (Dicentrarchus labrax) farms experiencing a high mortality rate. Diseased fish showed haemorrhages on the external body surface with exophthalmia, cataracts, scale desquamation, skin ulcers and fin erosions. The most common post-mortem findings were congested internal organs, particularly the liver and posterior kidney. Twenty-eight A. Veronii and 11 A. caviae isolates were identified biochemically by the Vitek 2 system and then confirmed by PCR and phylogenetic analysis. Hemolysin (hlyA) and aerolysin (aer) were the most abundant virulence genes in the recovered isolates, followed by cytotoxic enterotoxin (act) and heat-stable enterotoxin (ast). A. caviae was more virulent than A. veronii for D. labrax fingerlings as LD50 ranging between (>1 × 108 -6.2 × 107 ) for A. veronii and (2.9 × 107 -8.3 × 107 ) for A. caviae. The sensitivity test indicated the effectiveness of norfloxacin, doxycycline and oxytetracycline against the tested isolates. Serum cortisol significantly increased in the infected groups, while catalase and glutathione peroxidase activities significantly decreased at 2 days post-infection (DPI) and then increased at 6 DPI. The presence of virulence genes was associated with bacterial pathogenicity expressed in fish mortality rate. Virulence genes also drastically affect cortisol levels more than catalase and glutathione peroxidase levels.

Virulence genes contributing to Aeromonas hydrophila pathogenicity in Oreochromis niloticus.

Bacterial diseases are the main cause of high economic loss in aquaculture, particularly gram-negative bacteria. This study was conducted for the isolation and identification of Aeromonas and Pseudomonas spp. from diseased fish. Twenty-two Aeromonas and sixteen Pseudomonas isolates were recovered from diseased Nile tilapia (Oreochromis niloticus) raised in eight earthen ponds in Elhox, Metoubes, Kafrelsheikh, Egypt. The recovered isolates were further identified using PCR as 22 Aeromonas hydrophila, 11 Pseudomonas aeruginosa, and 5 Pseudomonas fluorescens isolates. The 22 A. hydrophila isolates were screened for the presence of four virulence genes. Sixteen of the isolates (72.72%) were positive for the aerolysin gene (aer); 4 (18.18%) harbored the cytotoxic enterotoxin gene (act); and 2 (9.09%) carried the hemolysin A gene (hylA) while the cytotonic heat-stable enterotoxin gene (ast) was absent from all the tested isolates. The pathogenicity test indicated the direct relationship between the mortality percentage and the genotype of the tested A. hydrophila isolates as the mortality rates were 63.3 and 73.3% for isolates with two virulence genes (aer+ & act+, and aer+ and hylA+, respectively), followed by 40, 53.3, and 56.6% for isolates with only one virulence gene (hylA, act, and aer, respectively) and 20% for isolates lacking virulence genes. Based on the sensitivity test, the multi-antibiotic resistance profiles were as follows: 90.9% of the A. hydrophila isolates were sensitive to florfenicol and doxycycline; then 68.18% were susceptible to oxytetracycline, norfloxacin, and ciprofloxacin; and 63.63% were susceptible to sulfamethoxazole-trimethoprim, while only 27.27 and 4.5% were sensitive to erythromycin and cephradine, respectively, and all the isolates were resistant to amoxicillin and ampicillin.