BACKGROUND: Recently chitosan nanoparticles have been applied as an immunostimulant and adjuvant in biomedical fields. OBJECTIVES: In this study the adjuvant effects of nanochitosan, derived from shrimp shell's chithin, on immunogenicity of Aeromonas hydrophila vaccine in common carp were evaluated. METHODS: Totally 300 juvenile common carp (51±5.6 g, Mean ±SD) were randomly divided into four groups each contained three subgroups with 25 fishes. Groups 1, 2 and 3 were immunized intraperitoneally with A. hydrophila vaccine. No adjuvant used in group 1, but nanochitosan and freund adjuvant were used with vaccine in groups 2 and 3 respectively. Group 4 (Control) injected with normal saline instead of vaccine. Fish were immunised at days zero and 14 and fishes were reared in the same situation for 6 weeks. Serum samples were taken every other weeks and immunological parameters including WBC, serum lysozyme and bactericidal activity, serum complement activity, NBT reduction activity, total protein and globulin levels and anti A.hydrophila antibody titer were compared among the groups. At the end of experiment all groups were challenged with LD50 concentration of A. hydrophila and mortality recorded for ten days. RESULTS: WBC, serum lysozyme and complement activity, NBT activity, globulin level and anti A.hydrophila antibody titer were increased in nanochitosan and freund groups compare to control group (P<0.05). Serum bactericidal activity and serum albumin level showed no significant change among the groups (P>0.05). Challenge mortality decreased in immunised groups compare to control. CONCLUSIONS: Nonochitosan posses proper adjuvant effects which is comparable with freund adjuvant and can be a good candidate as an alternative to current adjuvant in fish vaccines.

Introduction: In the present study, some of the commercial fish farms located in the Black Sea region of Turkey, were screened for bacteria between 2012 and 2014.Material and Methods: The bacterial agents isolated from fish were identified by classical biochemical tests and the rapid diagnostic tests (API 20 E and API 20 Strep). All strains were further identified by sequencing of the 16S rRNA genes. The strains were also investigated for resistance to different antimicrobials by the disc diffusion method. Antibiotic resistance genes, including tetracycline (B), β-lactam (ampC, blaTEM, blaPSE), florfenicol (floR), erythromycine (ereA, ereB), sulphonamide (sulI, sulII), and trimethoprim (dhfr1) genes, were determined by the PCR method.Results:Vibrio anguillarum, Vibrio fluvialis, Photobacterium damselae subsp. piscicida, Pseudomonas luteola, Lactococcus garvieae, Streptococcus iniae, Aeromonas hydrophila, and Yersinia ruckeri were isolated from marine and freshwater cultured fish. According to the results of disc diffusion, all isolates were sensitive to florfenicol, trimethoprim+sulfamethoxazole, oxitetracycline, and enrofloxacin, and resistant to lincomycin, penicillin G, and amoxicillin. Also, sulI, sulII, and floR resistance genes were detected in the bacteria.Conclusion: The results of the study open up the opportunity to perform further investigations which could determine the possible role of ARGs in fish pathogens.

Introduction: Immune-potentiating functions of Lactobacillus plantarum strains in the common carp were evaluated. Material and Methods: Fourteen days of feeding fish dry diet supplemented with the bacteria provided parameters of nonspecific humoral immunity (lysozyme, ceruloplasmin, γ-globulin, total protein levels, and serum bactericidal activity) and cellular immunity (pinocytosis, respiratory burst activity, and potential killing activity of organ phagocytes), as well as the proliferative response of organ lymphocytes stimulated with mitogens. The resistance of fish to infection with Aeromonas hydrophila was also determined. Results: Dietary supplementation with L. plantarum had a substantial influence on the activity of organ phagocytes, especially the potential killing activity of head kidney cells. A significant increase in the proliferative activity of LPS-stimulated B lymphocytes and in the levels of γ-globulins and total protein was observed. The supplemented diet conveyed higher resistance than the control diet as the cumulative fish mortalities after infection with A. hydrophila were 65% and 85%, respectively. Conclusion: The results indicate that dietary supplementation with L. plantarum stimulates the antibacterial resistance of common carp and may reinforce defence against bacterial infections, but further studies need to be conducted.

A study was conducted to investigate the pathogenicity and the median lethal dose (LD50) of Aeromonas hydrophila isolated from clinically diseased catfish against apparently healthy homologous fish to evaluate the lethality of extra-cellular products (ECPs) of the isolated strain in vivo. For pathogenicity experiment, five different concentrations of Aeromonas hydrophila strain BNS 01614 including 3× 108, 1.5 × 108, 1.5 × 107, 1.5 × 106 and 1.5 × 105 CFU/fish used via intra peritoneal. The results revealed that pathogenicity of BNS 01614 was confirmed by the mortality of 30 % to 100 % of all tested fish within 4 to 12 days with LD50 1.5 × 107 CFU/fish. The Concentrated extracellular products (ECPs) of the selected bacterium were prepared and confirmed to be toxic in Clarias garipineus with LC50 of 20µg.

Aeromonas hydrophila is a Gram negative, positive oxidase, anaerobic, and opportunistic bacteria that, under certain conditions, become a pathogen (in humans and fish). This bacterium causes toxin and host infection in which different antibiotic resistance in isolated strains has been reported in different regions of the world. The aim of this study was to determine the prevalence of this bacterium and its susceptibility to common antibiotics in Tabriz city.50 samples from 5 Reared Oncorhynchus mykiss farms in Tabriz city (For each farm,10 numbers) were randomly assigned to suspected fish to the disease. By using biochemical tests, 14 samples (28%) from 2 Fish farms (40%) were identified as A.hydrophila. Antibiogram for these specimens showed that the bacterium had the highest resistance tovancomycin (100 %) and clindamycin (92.8%) antibiotics, and has the most sensitivity to the antibiotics ofsultrim, tetracycline and oxytetracycline with 71.4%.Considering the different antibiotic resistance pattern in this study and other similar studies,the necessity of examining the pattern of resistance in each region seems necessary.

In the present study, Aeromonas (A.) hydrophila was isolated from a captive-bred adult freshwater stingray (Potamotrygon motoro) reared at a commercial aquarium in Korea. The stingray had bites on its fins, hemorrhages on the ventral part, and congested internal organs. A bacterium was isolated from kidney and subsequently identified as A. hydrophila. Based on phylogenetic analysis results, the isolate in the present study (SNUAh-LA1) was most closely related to A. hydrophila AH10 (China) and A. hydrophila AKR1 (Korea). It is most likely that the pathogen infection resulted from Potamotrygon motoro cohabiting with ricefish (Oryzias latipes).

Identification of microorganisms, including bacteria, are widely used especially in environmental studies, biotechnology, clinical microbiology, microbial forensics, and in research study. The conventional method of bacteria identification is based on phenotypic observation techniques by profiling an organism’s metabolic attributes or some aspect of its chemical composition. Then, interpretation of test results involves substantial subjective judgement. Currently, general 16S rRNA sequencing and specific PCR play an important role in the accurate and faster identification of bacteria. The aim of this study is to compare the identification of the genus or species of bacteria from agricultural waste using conventional microbiology biochemical test and molecular techniques PCR 16S rRNA universal amplified ribosomal region (UARR) sequencing. A total of 72 agricultural waste samples and 2 ATCC culture as positive control were tested. Out of two ATCC bacteria and fifteen bacteria isolates identified by the biochemical test, twelve species (71%) of bacteria gave exactly the same bacteria genus as the 16S rRNA sequencing results. Aeromonas hydrophilia, Alcaligenes faecalis and Acinetobacter calcoaceticus was revealed as Pseudomonas sp. from the sequencing results. As for Alcaligenes sp., the results from the sequencing is Stenotrophomonas maltophilia. Previous reports also showed different results of the same isolate which were from similar classification, and closely related to each other. The limited number of biochemical tests available in a laboratory will contribute to misidentification of a proposed specie.

Moribund albino catfish, Clarias sp., displayed from an indoor private commercial aquarium were submitted in the laboratory for diagnostic examination. Dense culture of bacteria was recovered from the kidney and was characterized using Vitek System 2 and showed 98% probability to Aeromonas (A.) hydrophila. PCR result showed positive using A. hydrophila extracellular hemolysin gene ahh1 (130 bp) and aerolysin gene aerA (309 bp). The 16S rRNA gene was identical and exhibited 97% sequence similarity with the other known isolates of A. hydrophila available in the GenBank. In this paper, we reported the isolation and molecular detection of A. hydrophila from an albino catfish.

Objective: The aerolysin (aerA) is a virulence indicator used to identify the pathogenicity of the Aeromonas strain. Targeting a pathogen&apos;s crucial virulence gene for detection is essential, as it determines the potential threat to the host. This study aimed to develop a gold nanoparticle (AuNP) probe for detecting the gene aerA in Aeromonas hydrophila among field samples. Materials and Methods: Kidney samples among both healthy and sick Nile tilapias in five provinces of Luzon Island were collected for bacterial analysis. Screening using specific primers targeting aerA was conducted in parallel with testing the AuNPs probe on the same sample set. The positive control provided by BFAR-NFLD, confirmed by polymerase chain reaction (PCR) assay, was used as a positive sample containing the target gene. Results: The AuNP probe demonstrated a computed accuracy of 81.32%, sensitivity of 100%, and specificity of 81.26%. Among the 257 reactions, 59 were false positives, while no false negative results were observed. The AuNP probe could detect aerA at levels as low as 30 ng/μl. The low prevalence of the target gene may be attributed to the use of general media instead of specific media like Rimler-Shotts agar. Conclusion: The established colorimetric detection method for A. hydrophila with the aerA gene offers a swift alternative to PCR, negating the requirement for advanced equipment like a thermal cycler. [J Adv Vet Anim Res 2023; 10(4.000): 593-598]

This study was undertaken for isolation, identification and determination of antibiogram profile of Aeromonas hydrophila in farm raised fresh water prawn (Macrobrachium rosenbergii) on five commercial ghers. Fresh water prawns (n=25) were collected from five ghers located at Satkhira, Bagerhat and Khulna districts of Bangladesh. Brain (n=25), muscle (n=25) and intestine (n=25) samples were collected aseptically from fresh water prawn and inoculated into alkaline peptone (APW) water for enrichment at 37℃ for 8 h. Enriched cultured was streaked into Thiosulfate Citrate Bile Salt Sucrose (TCBS) agar to isolate bacteria. Bacteria were identified by cultural, Gram staining, biochemical properties and polymerase chain reaction (PCR) assay. The antibiogram profiles of bacteria were investigated against 5 commonly used antibiotics (Gentamicin, Cefalexin, Ampicillin, Azithromycin and Ciprofloxacin) by disc diffusion method. Thirteen A. hydrophila isolates were identified and the prevalence of the A. hydrophila in fresh water prawn was 17.33%. All 13 (100%) isolates were sensitive to Ciprofloxacin, Gentamicin, Azithromycin and resistant to Ampicillin and Cefalexin. The results of this study indicate that farm raised fresh water prawn harbor multidrug resistant A. hydrophila which might causes public health problem if enter into human food chain.