-
Veterinary World Aug 2021The emergence of antibiotic-resistant bacterial pathogens has been increasingly reported, which has resulted in a decreasing ability to treat bacterial infections....
BACKGROUND AND AIM
The emergence of antibiotic-resistant bacterial pathogens has been increasingly reported, which has resulted in a decreasing ability to treat bacterial infections. Therefore, this study investigated the presence of spp., including its antibiotic resistance in various fish samples, spp., , and , obtained from Kelantan and Terengganu, Malaysia.
MATERIALS AND METHODS
In this study, 221 fish samples, of which 108 ( spp., n=38; , n=35; and , n=35) were from Kelantan and 113 ( spp., n=38; , n=35; and , n=40) were from Terengganu, were caught using cast nets. Then, samples from their kidneys were cultured on a Rimler Shott agar to isolate spp. Polymerase chain reaction (PCR) was used to confirm this isolation using specific gene primers for species identification. Subsequently, the isolates were tested for their sensitivity to 14 antibiotics using the Kirby-Bauer method, after which the PCR was conducted again to detect resistance genes: , -, , , , -, and .
RESULTS
From the results, 61 isolates were identified as being from the genus using PCR, of which 28 were , 19 were , seven were , and seven were . Moreover, 8, 12, and 8 of ; 4, 3, and 12 of ; 6, 0, and 1 of ; and 3, 3, and 1 of were obtained from spp., , and , respectively. In addition, the isolates showed the highest level of resistance to ampicillin (100%), followed by streptomycin (59.0%), each kanamycin and nalidixic acid (41.0%), neomycin (36.1%), tetracycline (19.7%), sulfamethoxazole (14.8%), and oxytetracycline (13.1%). Resistance to gentamicin and ciprofloxacin both had the same percentage (9.8%), whereas isolates showed the lowest resistance to norfloxacin (8.2%) and doxycycline (1.6%). Notably, all isolates were susceptible to chloramphenicol and nitrofurantoin. Results also revealed that the multiple antibiotic resistances index of the isolates ranged from 0.07 to 0.64, suggesting that the farmed fish in these areas were introduced to the logged antibiotics indiscriminately and constantly during their cultivation stages. Results also revealed that the gene was detected in 19.7% of the isolates, whereas the tetracycline resistance genes, and , were detected in 27.9% and 4.9% of the isolates, respectively. However, β-lactam resistance genes, and , were found in 44.3% and 13.1% of isolates, respectively, whereas and genes were found in 3.3% and 13.1% of the isolates, respectively.
CONCLUSION
This study, therefore, calls for continuous surveillance of antibiotic-resistant spp. in cultured freshwater fish to aid disease management and better understand their implications to public health.
PubMed: 34566322
DOI: 10.14202/vetworld.2021.2064-2072 -
Transboundary and Emerging Diseases Jul 2022Monogenean infection of the internal organs is extremely rare when compared to external infections. This study describes mass mortality of Nile tilapia (Oreochromis...
Monogenean infection of the internal organs is extremely rare when compared to external infections. This study describes mass mortality of Nile tilapia (Oreochromis niloticus L.) originating from co-infection with Enterogyrus spp. and Aeromonas jandaei following transport stress. The first fish deaths occurred on day 1 post-transport, while cumulative mortality reached approximately 90% by day 10 post-stocking. An atypical amount of pale (whitish) faeces floating on the surface of the water as well as typical clinical signs of motile Aeromonas septicemia, were reported. Adult monogeneans and countless eggs of monogeneans were found in the stomachs and the intestines of both moribund and dead fish, respectively. Two strains of A. jandaei were isolated from the kidneys. Scanning electron microscope microphotographs of the stomach revealed the presence of numerous monogeneans penetrating deep into the gastric tissue, and diffuse lesions filled with bacilliform bacteria. Histopathological examination showed multifocal eosinophilic infiltrate, gastric gland and epithelial necrosis with sloughed necrotic debris in the lumen. This is the first report of co-infection by Enterogyrus spp. and A. jandaei in Nile tilapia and the first report of Enterogyrus coronatus, Enterogyrus foratus, and Enterogyrus malbergi parasitizing tilapia in Brazil. These findings indicate that synergic co-infection by Monogenean stomach parasites (E. coronatus, E. foratus, and E. malbergi) and A. jandaei may induce high mortalities in tilapia following transport stress.
Topics: Aeromonas; Animals; Cichlids; Coinfection; Fish Diseases; Ovum; Tilapia; Trematoda
PubMed: 34406699
DOI: 10.1111/tbed.14295 -
Frontiers in Microbiology 2021The prevalence of -positive bacteria in 5,169 domestic animal-derived samples collected by USDA Food Safety and Inspection Service between October 2018 and May 2019 was...
The prevalence of -positive bacteria in 5,169 domestic animal-derived samples collected by USDA Food Safety and Inspection Service between October 2018 and May 2019 was investigated. A procedure including enriched broth culture and real-time PCR targeting to were used for the screening. Fifteen positive isolates were identified, including one plasmid-borne -positive strain, EC2492 (reported elsewhere) and 14 -positive strains from poultry (1), catfish (2), and chicken rinse (11) samples, resulting in an overall prevalence of -positive bacteria 0.29% in all meat samples tested. Analysis of 16S rRNA and whole genome sequences revealed that all 14 strains belonged to . Data from phylogenetic analysis of seven housekeeping genes, including , and , indicated that nine strains belonged to and five strains belonged to . Antimicrobial tests showed that almost all -positive strains exhibited high resistance to colistin with MICs ≥ 128mg/L, except for one strain, which showed a borderline resistance with a MIC of 2 mg/L. A segment containing two adjacent and lik genes was found in two and one strains and a variety of IS-like elements were found in the flanking regions of this segment. A -related lipid A phosphoethanolamine transferase gene was present in all 14 strains, while an additional -related lipid A phosphoethanolamine transferase gene was found in 5 strains only. In addition to genes, other antimicrobial resistance genes, including , , , , , , , , and were observed in chromosomes of some strains. The relative high prevalence of chromosome-borne genes and the close proximity of various IS elements to these genes highlights the need for continued vigilance to reduce the mobility of these colistin-resistance genes among food animals.
PubMed: 33995332
DOI: 10.3389/fmicb.2021.667406 -
Journal of Applied Microbiology Oct 2021This study aimed to evaluate the in vitro cytotoxicity and efficacy of synthetic host defence peptides (HDPs), alone or in combination with florfenicol (FFC),...
AIMS
This study aimed to evaluate the in vitro cytotoxicity and efficacy of synthetic host defence peptides (HDPs), alone or in combination with florfenicol (FFC), oxytetracycline (OTC) or thiamphenicol (TAP), against different pathogenic bacteria isolated from diseased fish.
METHODS AND RESULTS
Solid-phase synthesis, purification and characterization of several HDPs were performed manually, using the fluorenylmethyloxycarbonyl protecting group in different resins and via high-performance liquid chromatography-mass spectrometry, respectively. The in vitro cytotoxicity and antimicrobial activity of HDPs, FFC, OTC and TAP against Nile tilapia red blood cells (RBCs) and relevant fish pathogenic bacteria (Aeromonas, Citrobacter, Edwardsiella, Streptococcus, Lactococcus and Vibrio) was determined using the haemolysis assay and broth microdilution method, respectively. The checkerboard assay was used to evaluate the synergy between the most active HDPs and other antimicrobials against the tested strains. MUC 7 12-mer, FFC, OTC and TAP were not cytotoxic to Nile tilapia RBCs, in all tested concentrations. LL-37, (p-BthTX-I) and Hylin-a1 were not cytotoxic at concentrations up to 78·13, 19·53 and 9·77 μg ml , respectively. HDPs demonstrated potent antimicrobial activity (minimum inhibitory concentration ≤31·25 µg ml ) against Aeromonas jandaei (KR-12-a5), Citrobacter freundii (Kr-12-a5; (p-BthTX-I) ; LL-37; and Hylin a1), Streptococcus agalactiae (Hylin a1; (p-BthTX-I) and LL-37), Lactococcus garviae (Hylin a1), and Vibrio fluvialis (KR-12-a5). The combinations of (p-BthTX-I) with TAP and LL-37 with FFC showed synergistic activity against C. freundii (fractional inhibitory concentration index of 0·25 and 0·50, respectively).
CONCLUSIONS
Synthetic HDPs have the potential as a good treatment option for bacterial diseases in aquaculture.
SIGNIFICANCE AND IMPACT OF THE STUDY
The in vivo effectiveness of synthetic HDPs such as KR-12-a5; LL-37; (p-BthTX-I) and Hylin a1 can be tested alone or in combination with conventional antimicrobials as a treatment option to reduce the use of antimicrobials in aquaculture.
Topics: Aeromonas; Animals; Anti-Bacterial Agents; Anti-Infective Agents; Antimicrobial Cationic Peptides; Cichlids; Fish Diseases; Lactococcus; Microbial Sensitivity Tests; Vibrio
PubMed: 33742508
DOI: 10.1111/jam.15080 -
Scientific Reports Mar 2021Diseased Anabas testudineus exhibiting signs of tail-rot and ulcerations on body were collected from a fish farm in Assam, India during the winter season (November 2018...
Diseased Anabas testudineus exhibiting signs of tail-rot and ulcerations on body were collected from a fish farm in Assam, India during the winter season (November 2018 to January 2019). Swabs from the infected body parts were streaked on sterilized nutrient agar. Two dominant bacterial colonies were obtained, which were then isolated and labelled as AM-31 and AM-05. Standard biochemical characterisation and 16S rRNA and rpoB gene sequencing identified AM-31 isolate as Aeromonas hydrophila and AM-05 as Aeromonas jandaei. Symptoms similar to that of natural infection were observed on re-infecting both bacteria to disease-free A. testudineus, which confirmed their virulence. LC was determined at 1.3 × 10 (A. hydrophila) and 2.5 × 10 (A. jandaei) CFU per fish in intraperitoneal injection. Further, PCR amplification of specific genes responsible for virulence (aerolysin and enterotoxin) confirmed pathogenicity of both bacteria. Histopathology of kidney and liver in the experimentally-infected fishes revealed haemorrhage, tubular degeneration and vacuolation. Antibiotic profiles were also assessed for both bacteria. To the best of our knowledge, the present work is a first report on the mortality of farmed climbing perch naturally-infected by A. hydrophila as well as A. jandaei, with no records of pathogenicity of the latter in this fish.
Topics: Aeromonas; Aeromonas hydrophila; Animals; Fish Diseases; Gram-Negative Bacterial Infections; Hemorrhagic Septicemia; Perches
PubMed: 33712685
DOI: 10.1038/s41598-021-84997-x -
Antimicrobial Resistance and Infection... Feb 2021The study aimed to elucidate the species taxonomy, clinical manifestations, virulence gene profiles and antimicrobial susceptibilities of Aeromonas strains isolated from...
BACKGROUND
The study aimed to elucidate the species taxonomy, clinical manifestations, virulence gene profiles and antimicrobial susceptibilities of Aeromonas strains isolated from life-threatening bacteremia in southeastern China.
METHODS
Clinical samples of Aeromonas causing bacteremia were isolated from a teaching hospital in Wenzhou from 2013 to 2018 and a retrospective cohort study was performed. Aeromonas strains were identified at species level by housekeeping gene gyrB. Virulence and drug resistance-associated genes were screened by polymerase chain reaction (PCR) and antimicrobial susceptibility testing (AST) was performed by the VITEK 2 Compact system.
RESULTS
A total of 58 Aeromonas isolated from patients with bacteremia were collected during 6 years (2013-2018). 58 isolates were identified to five different species, where Aeromonas dhakensis appeared to be the predominant species (26/58), followed by Aeromonas veronii (13/58), Aeromonas caviae (10/58), Aeromonas hydrophila (7/58) and Aeromonas jandaei (2/58). 16 of 58 patients had poor prognosis. Poor prognosis was significantly associated with liver cirrhosis and inappropriate empirical antimicrobials therapy. The progression of bacteremia caused by Aeromonas was extremely fast, especially in A. dhakensis infections. Virulence genes aer, lip, hlyA, alt, ast, and act, were detected at ratios of 24.1% (14/58), 62.1% (36/58), 65.5% (38/58), 58.6% (34/58), 15.5% (9/58) and 65.5% (38/58), respectively. Antimicrobial susceptibility testing exhibited that 9 out of 58 isolates were identified as multi-drug resistant (MDR) organism. The bla gene was identified in all 9 MDR isolates. bla, bla, bla, bla, bla and aac(6')-Ib-cr were detected in 4 isolates, 2 isolates, 1 isolate, 3 isolates, 8 isolates, and 3 isolates, respectively. The majority of Aeromonas strains maintained susceptible to 3rd generation cephalosporins, aminoglycosides, fluoroquinolones and furantoin.
CONCLUSIONS
The prevalence and dangerousness of Aeromonas infections, especially A. dhakensis, are underestimated in clinic. Continuous monitoring is essential to keep track of MDR Aeromonas due to the increasing prevalence recently and a more effective measure is required to control the spread of resistance determinants.
Topics: Adult; Aeromonas; Aged; Anti-Bacterial Agents; Bacteremia; China; Female; Gram-Negative Bacterial Infections; Hospitals, Teaching; Humans; Male; Microbial Sensitivity Tests; Middle Aged; Phylogeny; Retrospective Studies; Virulence; Virulence Factors
PubMed: 33640019
DOI: 10.1186/s13756-021-00911-0 -
Microbial Drug Resistance (Larchmont,... Jul 2020Integrons are prokaryotic genetic elements known to carry and exchange antibiotic resistance gene cassettes through a site-specific recombinase called integrase. In this...
Integrons are prokaryotic genetic elements known to carry and exchange antibiotic resistance gene cassettes through a site-specific recombinase called integrase. In this work, 107 isolates from environmental origin, including fish, water, and sediments, were investigated for the presence of integrons. Using specific primers for Class 1, 2 and 3 integrases, only Class 1 and Class 2 integrons were detected. Detection of Class 2 integrases and their associated variable regions required two rounds of polymerase chain reaction (PCR). Sequencing of the 2 amplicons confirmed them as integrase-derived products. Class 1 integrons were detected in 26 out of 107 isolates. PCR amplification of the variable regions associated to these integrons revealed an outstanding homogeneity, 25 of them having variable regions with an identical A12-F-A2 cassette array and one integron carrying only the A16 cassette. To assess clone diversity, chromosomal DNA from isolates was subjected to enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR), which discarded clonality in all instances. Class 2 integrons were surprisingly more prevalent than Class1 integrons, being detected in 60 out of 107 isolates. Forty-six of them showed a unique ERIC profile, while the remaining 14 strains displayed profiles that could be grouped in five different patterns. Cassette arrangements of all Class 2 variable regions were those described as the most prevalent (A1-2-A1). A rather startling result of this work is the sensitivity to trimethoprim, streptomycin, and streptothricin of most strains, despite the presence of the cognate resistance genes. To know the integron distribution in environmental species, a phylogenetic reconstruction was done using D/B or D/A gene sequences. Isolates bearing these elements corresponded to , , , , , , , , , and sp. This work revealed an unusual high incidence of Class 2 integrons and a low variability of cassette arrangements in environmental species.
Topics: Aeromonas; Anti-Bacterial Agents; DNA, Bacterial; Drug Resistance, Multiple, Bacterial; Integrases; Integrons; Microbial Sensitivity Tests; Polymerase Chain Reaction
PubMed: 31990611
DOI: 10.1089/mdr.2019.0250 -
International Journal of Antimicrobial... Jan 2020Two novel phosphoethanolamine transferase genes, eptAv7 and eptAv3, were identified in the chromosome of an Aeromonas jandaei isolate from retail fish. The variants...
Two novel phosphoethanolamine transferase genes, eptAv7 and eptAv3, were identified in the chromosome of an Aeromonas jandaei isolate from retail fish. The variants showed 79.9% and 80.0% amino acid identity to MCR-7.1 and MCR-3.1, respectively, and increased colistin resistance 128- to 256-fold in Aeromonas salmonicida. The two variants with no mobile genetic element in the flanking regions were also observed in other Aeromonas species. This finding supports the view that Aeromonas is a reservoir for MCR-3 and MCR-7 mobile colistin resistance.
Topics: Aeromonas; Animals; Anti-Bacterial Agents; Aquaculture; Bacterial Proteins; Colistin; Drug Resistance, Bacterial; Ethanolaminephosphotransferase; Ethanolamines; Fish Diseases; Fishes; Humans; Phylogeny; Whole Genome Sequencing
PubMed: 31770630
DOI: 10.1016/j.ijantimicag.2019.11.013 -
Applied Microbiology and Biotechnology Nov 2019Streptococcus agalactiae is a major pathogen causing streptococcosis. To prevent and control this bacterial disease, antagonistic bacteria have become a new research...
Bacillus velezensis LF01: in vitro antimicrobial activity against fish pathogens, growth performance enhancement, and disease resistance against streptococcosis in Nile tilapia (Oreochromis niloticus).
Streptococcus agalactiae is a major pathogen causing streptococcosis. To prevent and control this bacterial disease, antagonistic bacteria have become a new research hotspot. This study evaluated the probiotic potential of Bacillus velezensis LF01 strain, which is antagonistic to S. agalactiae. The active compounds produced by LF01 showed antimicrobial activity against a broad spectrum of fish pathogens, including S. agalactiae, Streptococcus iniae, Aeromonas hydrophila, Edwardsiella tarda, Edwardsiella ictaluri, Aeromonas schubertii, Aeromonas veronii, Aeromonas jandaei, and Vibrio harveyi. The antimicrobial compounds were heat stable, pH stable, UV stable, resistant to proteases, and could be stored for a long time. To evaluate the probiotic function of LF01 in Nile tilapia, juveniles were divided into three treatment groups: a control group, an interval feeding group, and a continuous feeding group. Tilapia fed with LF01-supplemented diets (1.0 × 10 CFU/g) showed significantly better growth performances than those of the control group (P < 0.05). Tilapia fed with LF01-supplemented diets significantly increased lysozyme (LZY) and superoxide dismutase (SOD) activities. The expression of three immune-related genes (C3, lyzc, and MHC-IIβ) was higher in the intestine, head kidney, and gill of tilapia from the continuous feeding group than in those from the control group (P < 0.05). Tilapia fed with LF01-supplemented diets showed remarkably improved survival rates after S. agalactiae infection, and analysis of their intestinal tract pathogens revealed that the abundance of Edwardsiella and Plesiomonas had significantly decreased compared with the control group. Our findings demonstrate that LF01 is an effective antagonist against various fish pathogens and has potential for controlling infections by Streptococcus spp. and other pathogens in tilapia.
Topics: Animals; Antibiosis; Bacillus; Biological Control Agents; Cichlids; Fish Diseases; High-Throughput Nucleotide Sequencing; Probiotics; Streptococcal Infections; Streptococcus agalactiae
PubMed: 31654082
DOI: 10.1007/s00253-019-10176-8 -
Water Research Sep 2019The wide application of chlorine disinfectant for drinking water treatment has led to the appearance of chlorine-resistant bacteria, which pose a severe threat to public...
The wide application of chlorine disinfectant for drinking water treatment has led to the appearance of chlorine-resistant bacteria, which pose a severe threat to public health. This study was performed to explore the physiological-biochemical characteristics and environmental influence (pH, temperature, and turbidity) of seven strains of chlorine-resistant bacteria isolated from drinking water. Ozone disinfection was used to investigate the inactivation effect of bacteria and spores. The DNA concentration and cell surface structure variations of typical chlorine-resistant spores (Bacillus cereus spores) were also analysed by real-time qPCR, flow cytometry, and scanning electron microscopy to determine their inactivation mechanisms. The ozone resistance of bacteria (Aeromonas jandaei < Vogesella perlucida < Pelomonas < Bacillus cereus < Aeromonas sobria) was lower than that of spores (Bacillus alvei < Lysinibacillus fusiformis < Bacillus cereus) at an ozone concentration of 1.5 mg/L. More than 99.9% of Bacillus cereus spores were inactivated by increasing ozone concentration and treatment duration. Moreover, the DNA content of Bacillus cereus spores decreased sharply, but approximately 1/4 of the target genes remained. The spore structure exhibited shrinkage and folding after ozone treatment. Both cell structures and gene fragments were damaged by ozone disinfection. These results showed that ozone disinfection is a promising method for inactivating chlorine-resistant bacteria and spores in drinking water.
Topics: Chlorine; Disinfectants; Disinfection; Drinking Water; Ozone; Spores, Bacterial; Water Purification
PubMed: 31158616
DOI: 10.1016/j.watres.2019.05.014