Diversity of Antibiotic-resistant of Tentative Motile Aeromonas Species Isolated From Clarias gariepinus (Burchell 1822) Cultured in Earthen Ponds
Deborah Arimie AdahLawal Sa’iduSonnie Joshua OniyeM. A. RajiOluwafemi Babatunde DaoduAdakole Sylvanus Adah
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Background: Aeromonas species is one of the most important causes of diseases in Clarias gariepinus, a public health threat with significant economic losses. Objectives: In this research, the prevalence and variety of Aeromonas species isolated from C. gariepinus cultured in an earthen pond were investigated, as well as the antibiogram and multiple antibiotic resistance index Methods: Aeromonas species were isolated by culture and biochemical test and confirmed using a Microbact 24E kit. The antibiotic susceptibility to 10 antibiotics was determined using the Kirby-Bauer disk diffusion method. Results: Aeromonas species were isolated with a prevalence of 43.1% with 4 different phenospecies with the highest prevalence of 46(24 %) for Aeromonas hydrophila followed by 15(7.8%) for Aeromonas caviae, then 10(5.2%) for Aeromonas veronii sobria, and the least was observed for Aeromonas veronii veronii 6(3.1%). The Aeromonas species showed high resistance to amoxicillin, ampicillin, colistin sulfate, oxytetracycline, trimethoprim/sulfamethoxazole, and penicillin, with varying resistant patterns, and the multiple antibiotic resistance index values ranged between 0.20 and 0.80. Conclusion: There was a diversity of Aeromonas species associated with multiple antibiotic-resistant leading to the wide spread of antimicrobial resistance. Therefore, there is a need to control the use of antibiotics and ensure the effective use of biosecurity and preventive management measures in fish farms.Keywords:
Aeromonas veronii
Clarias gariepinus
Trimethoprim
Colistin
Sulfamethoxazole
Ten strains of Aeromonas spp. using a selective media Aeromonas agar base were isolated from the water, sediment and intestine content of healthy fish in fish ponds as well as the mud sample taken from rice fields, in Zhongshan city, Guangdong Province. They were identified by means of conventional physiological and biochemical tests together with 16S rDNA sequencing. In addition, the phylogenetic relationship between these strains and some type strains of Aeromonas spp. was analyzed based on their 16S rDNA sequences. Aeromonas hydrophila was found to be the most frequently isolated Aeromonas species as expected. Other isolates were identified as Aeromonas caviae, A. jandaei, A. veronii, respectively, among which A. jandaei and A. veronii were new record of Aeromonas species in China. This is the first study for the diversity of Aeromonas species in the environment in our country.
Aeromonas veronii
Aeromonas caviae
Isolation
Aeromonas salmonicida
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Genotypic characterization of 215 Aeromonas strains (143 clinical, 52 environmental, and 20 reference strains) showed that Aeromonas aquariorum (60 strains, 30.4%) was the most frequently isolated species in clinical and water samples and could be misidentified as Aeromonas hydrophila by phenotypic methods.
Aerolysin
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Aeromonas is a kind of bacteria which could infect human and aquatic animals.One Aeromonas strain of pathogenic bacteria HBJY01,isolated from diseased Trionyx sinensis,was identified through molecular method.Phylogenetics status of Aeromonas(Aeromonadaceae) was analyzed.Using the 16S rRNA and gyrB genes,the Aeromonas strain was identified,bacterial phylogenetics trees were constructed and pairwise distance was calculated.Based on these,phylogenetics status of Aeromonas(Aeromonadaceae) was analyzed.HBJY01 was very similar to Aeromonas veronii in terms of 16S rRNA and gyrB gene sequences.In the genetic distance,Aeromonas and other Vibrionaceae had great differences.HBJY01 was identified as Aeromonas veronii.Aeromonas should be an independent new family,Aeromonadaceae,not be a genus of Vibrionaceae.
Aeromonas veronii
Aeromonas caviae
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Abstract. Mulia DS, Utomo T, Isnansetyo A. 2022. The efficacy of Aeromonas hydrophila GPl-04 feed-based vaccine on African catfish (Clarias gariepinus). Biodiversitas 23: 1505-1510. Aeromonas hydrophila is a pathogenic bacterium to African catfish (Clarias gariepinus). Vaccination is one of the strategic efforts to control this type of bacterial attack. The objective of this study was to determine the effectiveness of A. hydrophila GPl-04 feed-based vaccine for African catfish. This study used A. hydrophila GPl-04 strain isolate as vaccine material. Sample used was African catfish measuring 10-13 cm length, weighing 16-25 g, taken from aquaculture ponds in the Purbalingga area, Central Java, Indonesia. This study applied experiments under the completely randomized design (CRD) method with 5 treatments and 3 replications. The treatments consisted of P0: feed without vaccine (control); P1: feed with 10 mL/100 g dose of vaccine given for 10 days; P2: feed with 10 mL/100 g dose of vaccine given for 15 days; P3: feed with 15 mL/ 100 g dose of vaccine given for 10 days; and P4: feed with 15 mL/ 100 g dose of vaccine given for 15 days. The challenge test was carried out on week 3, by injecting 0.1 mL of A. hydrophila suspension at 105CFU/mL dose per fish. The main parameters included the fish antibody titer, survival rate (SR), relative percent survival (RPS), mean time to death (MTD), and growth rate. Supporting parameters included water quality such as water temperature, water pH, and dissolved O2 levels. The main parameter data were analyzed using analysis of variance (ANOVA) and Duncan multiple range test (DMRT) at 5% test level, and the supporting parameter was descriptively quantitative analyzed. The results showed A. hydrophila GPl-04 feed-based vaccine could increase antibody titer, SR, RPS, and MTD of African catfish (P<0.05). In addition, vaccination did not adversely affect the growth. The A. hydrophila GPl-04 feed-based vaccine was effective in protecting African catfish from A. hydrophila attack where 10 mL/100 g dose, given for 10 days (P1) was indicated as the most effective compared to other vaccination treatments.
Clarias gariepinus
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Aeromonas is one of the major pathogens which frequently cause the bacterial diseases in freshwater fish terrapin and crustaceans.Aeromonas hydrophila caused economic losses widely.In order to detect Aeromonas spp.and Aeromonas hydrophila,two pairs of specific primers were designed according to the glycerophospholipid-cholesterol acyltransferase(GCAT) gene of Aeromonas spp.and the 16S rRNA gene of Aeromonas hydrophila.So a duplex PCR assay was developed to amplify the 16S rRNA gene and the GCAT gene.The reaction conditions of the duplex PCR were optimized and PCR products were sequenced.Meanwhile,the specificity of duplex PCR was studied.Five Aeromonas hydrophila strains,seven Aeromonas strains and the other five strains of non Aeromonas were tested by duplex PCR.The results showed that the GCAT gene and the 16S rRNA gene could be amplified from Aeromonas hydrophila strains,but only the GCAT gene from the other Aeromonas species and neither from the other five strains.It could be concluded that the duplex PCR was specific and could be used in quick diagnose Aeromonas hydrophila and Aeromonas spp.
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The high diversity of Aeromonas spp. results in various pathogenicity levels. This group of bacteria causes a serious disease named motile Aeromonas septicemia (MAS) in catfish (Clarias spp.). This study aimed to characterize the species and virulence gene diversity of Aeromonas spp. isolated from diseased catfish.Nine Aeromonas spp. were isolated from infected catfish cultivated in Java, Indonesia, and they were identified at the phenotypic and molecular levels (16S rDNA). The virulence genes assessed included aer/haem, alt, ast, flaA, lafA, and fstA.Phylogenetic analysis identified nine isolates of Aeromonas spp.: Aeromonas hydrophila (11.11%), Aeromonas caviae (11.11%), Aeromonas veronii bv. veronii (44.44%), and Aeromonas dhakensis (33.33%). Virulence genes, such as aer/haem, alt, ast, flaA, lafA, and fstA, were detected in all isolates at frequencies of approximately 100%, 66.67%, 88.89%, 100%, 55.56%, and 66.67%, respectively. This study is the first report on A. dhakensis recovered from an Indonesian catfish culture. Furthermore, our study revealed the presence of A. veronii bv veronii, a biovar that has not been reported before in Indonesia.This finding confirms that MAS was caused by multiple species of Aeromonas, notably A. dhakensis and A. veronii bv veronii, within Indonesian fish culture. The presence of these Aeromonas species with multiple virulence genes poses a significant threat to the freshwater aquaculture industry.
Aeromonas veronii
Aerolysin
Aeromonas caviae
Biovar
Francisella
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Aeromonas hydrophila感染例は稀ではあるが致死的となることがある．本感染症の3例を経験したので報告する．症例1:79歳，男性．胆嚢癌に胆嚢胆管切除を施行．翌日，頻脈となり血圧が低下した．白血球，Hb，血小板が低下，皮下出血が広がりDICとなった．原因不明の感染症を疑い治療を行ったが全く反応なく術後52時間で死亡した．死亡後，痰，胃液，ドレーン排液，動脈血，創浸出液からAeromonas hydrophilaが検出された．症例2:73歳，男性．食道癌に右開胸胸部食道全摘，食道瘻，胃瘻造設を施行した．2POD腹膜炎を併発，再手術を施行した．創感染の培養でAeromonasと判明した．創を開放し抗菌剤を投与した．創感染は軽快したが最終的に在院死した．症例3:72歳，男性．S状結腸線維腫を切除した．創感染の培養でAeromonasと判明した．ドレナージとPZFXで軽快した．外科手術の際には本感染症の存在と危険性を念頭に置いた上で適切なシステムの構築と迅速な対応が必要である．
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Bacterial pathogens are the most serious agents causing diseases in both wild and cultured fish resulted in massive mortalities and economic losses. Motile Aeromonas Septicemia (MAS) is a prevalent bacterial disease caused by Aeromonas hydrophila (A. hydrophila) that impacts freshwater fish. This research aimed to evaluate doxycycline (DOX) antibacterial activity against A. hydrophila both in vitro and in vivo. The minimum inhibitory concentration (MIC) and mutant prevention concentration (MPC) of DOX against A. hydrophila previously isolated from African catfish (Clarias gariepinus) were determined to be 0.78µg/mL and 3.9μg/mL, respectively. For in vivo experiment, a total number of 80 apparently healthy African catfish, were distributed randomly into four equal groups. Group 1 (non-infected, non-treated) was kept as control, Group 2 (non-infected and treated) was non-infected and treated with 20 mg/Kg BW of DOX for 5 successive days in feed, Group 3 (infected) was inoculated intraperitoneally (IP) with A. hydrophila (2× 108 CFU/ mL) and Group 4 (infected and treated) was infected with A. hydrophila then treated with 20 mg DOX/Kg BW. Our results revealed 70% mortality in African catfish experimentally challenged with A. hydrophila (Group 3). Moreover, significant elevation of serum alanine aminotransferase (ALT) (89±16.26, 54.67±6.44, 36±5.29 U/L, respectively), aspartate aminotransferase (AST) (195±7.64, 221.33±17.9, 211.33±12.72 U/L, respectively) and creatinine (0.68±0.098, 0.76±0.052, 0.58 ±0.023 mg/dL, respectively) was observed on 1st, 7th and 14th days post treatment. While treatment of the infected fish (Group 4) with DOX decreased the mortality rate to 30 %, improved the clinical signs and significantly reduced serum ALT (30.67±6.01, 22.67±1.86 U/L, respectively) and AST (153±7.57, 147.67±6.7 U/L, respectively) on 7th and 14th days post treatment. Also, it significantly decreased creatinine (0.21±0.026, 0.25±0.047, 0.21±0.053 mg/dL, respectively) levels at 1st, 7th and 14th days post treatment when compared with those of Group 3. The results showed that DOX could be used as an effective treatment against A. hydrophila infection in African catfish with little adverse effects.
Clarias gariepinus
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