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MicrobiologyOpen Nov 2019Aeromonas is recognized as a human pathogen following ingestion of contaminated food and water. One major problem in Aeromonas identification is that certain species are...
Proteomic characterization and discrimination of Aeromonas species recovered from meat and water samples with a spotlight on the antimicrobial resistance of Aeromonas hydrophila.
Aeromonas is recognized as a human pathogen following ingestion of contaminated food and water. One major problem in Aeromonas identification is that certain species are phenotypically very similar. The antimicrobial resistance is another significant challenge worldwide. We therefore aimed to use mass spectrometry technology for identification and discrimination of Aeromonas species and to screen the antimicrobial resistance of Aeromonas hydrophila (A. hydrophila). A total of 150 chicken meat and water samples were cultured, and then, the isolates were identified biochemically by the Vitek 2 Compact system. Proteomic identification was performed by MALDI-TOF MS and confirmed by a microchannel fluidics electrophoresis assay. Principal component analysis (PCA) and single-peak analysis created by MALDI were also used to discriminate the Aeromonas species. The antimicrobial resistance of the A. hydrophila isolates was determined by Vitek 2 AST cards. In total, 43 samples were positive for Aeromonas and comprised 22 A. hydrophila, 12 Aeromonas caviae (A. caviae), and 9 Aeromonas sobria (A. sobria) isolates. Thirty-nine out of 43 (90.69%) Aeromonas isolates were identified by the Vitek 2 Compact system, whereas 100% of the Aeromonas isolates were correctly identified by MALDI-TOF MS with a score value ≥2.00. PCA successfully separated A. hydrophila, A. caviae and A. sobria isolates into two groups. Single-peak analysis revealed four discriminating peaks that separated A. hydrophila from A. caviae and A. sobria isolates. The resistance of A. hydrophila to antibiotics was 95.46% for ampicillin, 50% for cefotaxime, 45.45% for norfloxacin and pefloxacin, 36.36% for ceftazidime and ciprofloxacin, 31.81% for ofloxacin and 27.27% for nalidixic acid and tobramycin. In conclusion, chicken meat and water were tainted with Aeromonas spp., with a high occurrence of A. hydrophila. MALDI-TOF MS is a powerful technique for characterizing aeromonads at the genus and species levels. Future studies should investigate the resistance of A. hydrophila to various antimicrobial agents.
Topics: Aeromonas; Aeromonas caviae; Aeromonas hydrophila; Animals; Anti-Bacterial Agents; Bacterial Proteins; Bacterial Typing Techniques; Chickens; Drug Resistance, Bacterial; Humans; Meat; Microbial Sensitivity Tests; Proteome; Proteomics; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Water Microbiology
PubMed: 30614207
DOI: 10.1002/mbo3.782 -
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 -
Annals of Agricultural and... Jun 2023The aims of this study were to search for the presence of bacteria in sea snails () by using culturomics and Matrix-assisted laser desorption/ionization time-of-flight...
INTRODUCTION AND OBJECTIVE
The aims of this study were to search for the presence of bacteria in sea snails () by using culturomics and Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and the antibiotic resistance/susceptibility of the sea snails.
MATERIAL AND METHODS
The anti-microbial susceptibilities of Gram-negative bacteriawas assessed by the Kirby-Bauer disk diffusion method, the presence of the genes (mcr-1 to -5), the major carbapenemase and β-lactamase resistant genes in Gram-negative bacteria, using mPCR method and 16S rRNA sequence analysis of isolates.
RESULTS
Bacterial growth accounted for 100% and 94.2% in the samples of intestine and meat, respectively, in the snails. The main organisms identified by MALDI-TOF MS were subsp. salmonicida at 33.7%, followed by at 9.6% (10/104) and at 7.7% in meat and intestine samples. and are intrinsic or chromosomally-mediated resistant against ampicillin. No genes (-1 to -5), the major carbapenemase and β-lactamase resistant genes were found. subsp. showed very low levofloxacin and meropenem resistance levels at 2.9%. When the sequence was searched in the Blast database, the genome of isolate showed high similarity with the sequences.
CONCLUSIONS
Conclusions. The findings obtained not only provide data about the proportion of bacteria in the gut and meat of the sea snails and their antibiotic resistance/susceptibility, but also show the absence of carbapenemase, colistin, and β-lactamase resistant genes among bacterial isolates from sea snail gut microbes.
Topics: Animals; Anti-Bacterial Agents; RNA, Ribosomal, 16S; Drug Resistance, Bacterial; Anti-Infective Agents; Snails
PubMed: 37387372
DOI: 10.26444/aaem/163582 -
PloS One 2020Cholera remains a major global public health threat and continuous emergence of new Vibrio cholerae strains is of major concern. We conducted a molecular epidemiological...
Cholera remains a major global public health threat and continuous emergence of new Vibrio cholerae strains is of major concern. We conducted a molecular epidemiological study to detect virulence markers and antimicrobial resistance patterns of V. cholerae isolates obtained from the 2012-2015 cholera outbreaks in Ghana. Archived clinical isolates obtained from the 2012, 2014 and 2015 cholera outbreaks in Ghana were revived by culture and subjected to microscopy, biochemical identification, serotyping, antibiotic susceptibility testing, molecular detection of distinct virulence factors and Multi-Locus Variable-Number of Tandem-Repeat Analysis (MLVA). Of 277 isolates analysed, 168 (60.6%) were confirmed to be V. cholerae and 109 (39.4%) isolates constituted other bacteria (Escherichia coli, Aeromonas sobria, Pseudomonas aeruginosa, Enterobacter cloacae and Enterococci faecalis). Serotyping the V. cholerae isolates identified 151 (89.9%) as Ogawa, 3 (1.8%) as Inaba and 14 (8.3%) as non-O1/O139 serogroup. The O1 serogroup isolates (154/168, 91.7%) carried the cholera toxin ctxB gene as detected by PCR. Additional virulence genes detected include zot, tcpA, ace, rtxC, toxR, rtxA, tcpP, hlyA and tagA. The most common and rare virulence factors detected among the isolates were rtxC (165 isolates) and tcpP (50 isolates) respectively. All isolates from 2014 and 2015 were multidrug resistant against the selected antibiotics. MLVA differentiated the isolates into 2 large unique clones A and B, with each predominating in a particular year. Spatial analysis showed clustering of most isolates at Ablekuma sub-district. Identification of several virulence genes among the two different genotypes of V. cholerae isolates and resistance to first- and second-line antibiotics, calls for scaleup of preventive strategies to reduce transmission, and strengthening of public health laboratories for rapid antimicrobial susceptibility testing to guide accurate treatment. Our findings support the current WHO licensed cholera vaccines which include both O1 Inaba and Ogawa serotypes.
Topics: Anti-Bacterial Agents; Cholera; Cholera Toxin; Disease Outbreaks; Drug Resistance, Multiple, Bacterial; Ghana; Humans; Microbial Sensitivity Tests; Phylogeny; Serogroup; Tandem Repeat Sequences; Vibrio cholerae; Virulence
PubMed: 32649692
DOI: 10.1371/journal.pone.0236016 -
Animals : An Open Access Journal From... Feb 2023Plant extracts are a phytochemically-rich alternative to antibiotic and synthetic feed additives, with high systemic bioactivity in animals. The present study aimed to...
Plant extracts are a phytochemically-rich alternative to antibiotic and synthetic feed additives, with high systemic bioactivity in animals. The present study aimed to evaluate the effect of a hydroalcoholic extract of custard apple () leaf (ASLE) on the growth, hematobiochemical parameters, digestive enzyme activities, redox status, nonspecific immune response, and cold and bacterial infection tolerance in Nile tilapia (). A total of 300 Nile tilapia fingerlings (11.87 ± 0.48 g) were fed ASLE-supplemented diets at increasing levels of 0, 5, 10, 15, and 20 g/kg for 60 days. At the end of the feeding period, the fish were experimentally challenged with cold water stress or , and mortalities were recorded for 10 days. The results revealed that the growth performance and feed conversion ratio were significantly improved with an increasing level of ASLE supplementation. The hematologic profile and hepato-renal functions were retained within a healthy range in the various groups supplemented with an ASLE diet. Antioxidant status was significantly improved in the serum of fish fed ASLE-supplemented diets, in terms of superoxide dismutase (SOD), catalase (CAT) activities, reduced glutathione, and total antioxidant capacity. Meanwhile, the myeloperoxidase (MPO) and malondialdehyde (MDA) levels decreased significantly. Similarly, there was a noticeable improvement in the hepatic CAT and SOD activities and a reduction of hepatic MDA. Marked improvements in lysozyme activity, nitric oxide production, complement3 level, and phagocytic activity were recorded in groups fed ASLE-supplemented diets, which peaked with the 20 g ASLE/kg diet. Moreover, the serum glucose and cortisol levels significantly declined in groups fed ASLE at levels of 15-20 g/kg compared to the other groups. Supplementation with ASLE increased the activities of protease, lipase, and α-amylase. ASLE supplementation at a concentration of 10-20 g/kg diet enhanced the resistance of Nile tilapia to infection. According to this study, ASLE supplementation enhanced the antioxidant balance, non-specific immune response, physiological status, resistance against infection, and growth performance of Nile tilapia at supplementation levels of 10-20 g/kg diet.
PubMed: 36830534
DOI: 10.3390/ani13040746 -
Acta Veterinaria Scandinavica Jan 2020Antimicrobial resistant bacteria are emerging biological contaminants of the environment. In aquatic ecosystems, they originate mainly from hospitals, livestock manure...
BACKGROUND
Antimicrobial resistant bacteria are emerging biological contaminants of the environment. In aquatic ecosystems, they originate mainly from hospitals, livestock manure and private households sewage water, which could contain antimicrobial agents and resistant microorganisms. Aeromonas spp. occur ubiquitously in aquatic environments and they cause disease in fish. Motile aeromonads are also associated with human gastrointestinal and wound infections and fish can act as a transmission route of antimicrobial resistance (AMR) aeromonads to humans. The environmental ubiquity, the natural susceptibility to antimicrobials and the zoonotic potential of Aeromonas spp. make them optimal candidates for studying the AMR in aquatic ecosystems.
RESULTS
The AMR patterns of 95 motile aeromonads isolated from freshwater fish during 2013 and 2016 were analyzed. All samples from fish came from farms and natural water bodies located in northern Italy, which is an area characterized by high anthropic impact on the environment. The isolates were biochemically identified as Aeromonas hydrophila, Aeromonas sobria or Aeromonas caviae and AMR was determined by the standard disk diffusion method. All isolates were resistant to cloxacillin, spiramycin and tilmicosin. High AMR frequencies (> 95%) were detected for tylosin, penicillin and sulfadiazine. AMR to danofloxacin, enrofloxacin, flumequine, ceftiofur, aminosidine, colistin, doxycycline, gentamicin, marbocyl and florfenicol was observed at low levels (< 10%). No AMR to cefquinome was found. Logistic regression showed several differences in antimicrobial activity between complexes. According to the source of aeromonads, only few differences in AMR between isolates from farmed and wild fish were observed.
CONCLUSIONS
Our data revealed an increasing trend of AMR to neomycin and apramycin among Aeromonas isolates during the study period, while resistance to erythromycin, tetracycline and thiamphenicol decreased. All isolates were multidrug resistance (MDR), but A. caviae showed the highest number of MDR per isolate. In most isolates, various degrees of MDR were detected to macrolides, quinolones, fluoroquinolones, polymyxins and cephalosporins (third and fourth generations), which are listed, by the World Health Organisation, to be among the highest priority and critically important antimicrobials in human medicine. Our findings underlined that freshwater fish can act as potential source of MDR motile aeromonads. Due to their zoonotic potential, this can pose serious threat to human health.
Topics: Aeromonas; Animals; Anti-Bacterial Agents; Drug Resistance, Bacterial; Fish Diseases; Fisheries; Fresh Water; Gram-Negative Bacterial Infections; Italy
PubMed: 31973764
DOI: 10.1186/s13028-020-0504-y -
Animals : An Open Access Journal From... Dec 2020In recent times, nutraceuticals have been used extensively to identify promising feed additives for the improvement of the aquaculture industry through the enhancement...
In recent times, nutraceuticals have been used extensively to identify promising feed additives for the improvement of the aquaculture industry through the enhancement of growth and survival rates, potentiation of the immune responses, and fortification of the resistance against infectious bacterial diseases. In this study, Nile tilapia () were fed with diets supplemented with quinoa seeds (QU) or prickly pear fruit peel (PP) at the dose levels of 10% or 20% of the diet. After 45 days of the feeding trial, the fish were exposed to challenge. The pre-challenge indices indicated that both supplements mediated a significant improvement in most of the estimated parameters, including survival rate, antioxidant status, hematological and immunological indices, and hepatoprotective potential. These effects were recorded in the groups fed with high doses of the supplements (20%). The least changes were observed in the QU-supplemented fish. In the spleen tissue, the gene was upregulated in the PP-, PP- and QU-supplemented groups, while the expression of the gene remained unaffected in all the supplemented groups, except for the PP-supplemented group, which showed an upregulation. After the challenge with , the relative survival percentage was improved by the supplementation of PP and QU, particularly in the PP-supplemented group, possibly via the promotion of immunological responses, hepatoprotective potency, and modulation of the studied genes. Moreover, the morphological structure of the tissues showed marked recovery. The findings suggest that Nile tilapia fed with different levels of PP peel and QU seeds, particularly at the level of 20%, enhanced the immune response in fish and improved their resistance against infection.
PubMed: 33271917
DOI: 10.3390/ani10122266 -
Veterinary World Jan 2022Antibiotic resistance has been a progressively documented problem, resulting in treatment failure in humans and animals. This study aimed to investigate the...
BACKGROUND AND AIM
Antibiotic resistance has been a progressively documented problem, resulting in treatment failure in humans and animals. This study aimed to investigate the antimicrobial susceptibility and virulence of extensively drug-resistant (XDR) spp. in wild and its surrounding seawater along the coastal road of Port Said, Egypt.
MATERIALS AND METHODS
Specimens were examined bacteriologically, confirmed biochemically, and tested for their sensitivity against 11 antimicrobial agents. Molecular confirmation of the obtained isolates by was performed, followed by the detection of antimicrobial resistance and virulence genes.
RESULTS
spp. was recovered from fish (44%) and water samples (36%). was the most prevalent identified strain, followed by , , and . Moreover, 90% of the tested isolates were multidrug-resistant (MDR), while 26.67% were XDR. Tested isolates were resistant to b-lactams and sulfonamides (100%), oxytetracycline (90%), and streptomycin (62.22%) but completely susceptible to cefotaxime. XDR isolates successfully amplified resistance genes (, and ()) but not the () gene, although there was phenotypic resistance to streptomycin on plates. All XDR isolates carry the cytotoxic enterotoxin gene (), but gene was detected in only one isolate (12.5%).
CONCLUSION
Data in this study provide a recent update and highlight the role of wild mullet and seawater as reservoirs for MDR and XDR spp. that may pose a risk to humans as food-borne infection or following direct contact.
PubMed: 35369605
DOI: 10.14202/vetworld.2022.55-64 -
BMC Veterinary Research Dec 2022Bacterial pathogens are a great threat to fish production. Gram-negative bacteria are among the major bacterial fish pathogens and are zoonotic with the potential to...
Isolation and identification of major bacteria from three Ethiopian rift valley lakes live and processed fish, and water samples: implications in sanitary system of fish products.
Bacterial pathogens are a great threat to fish production. Gram-negative bacteria are among the major bacterial fish pathogens and are zoonotic with the potential to infect humans. This cross-sectional study was conducted to isolate and identify major gram-negative bacteria from live and processed fish, and water samples from Lakes Hawassa, Langanoo and Ziway. A total of 674 different types of samples: 630 tissue samples (210 samples for each intestine, Kkidney and liver collected from 210 live fish (Oreochromis niloticus, Cyprinus carpio and Clarias gariepinus), 20 processed fish samples from lake Ziway fish processing center and 24 lake water samples were included in the study from each lake. The mean values of pH, temperature, dissolved oxygen and nitrate in all water samples were within the normal range at which most freshwater fish species become non-stressed. Of a total of 674 samples included in the study, bacteria were isolated from 154(22.8%) samples with significant difference (P < 0.05) observed in some isolates with respect to sample origin. Of these 154 isolates, 103(66.8%) isolates were gram-negative bacteria consisting of 15 species based on morphology and a range of biochemical tests. From live fish samples, Escherichia coli was the dominant species with 15 isolates followed by Edwardsiella tarda (12), Salmonella Paratyphi (10), Salmonella Typhi (9), Shigella dysenteriae (7), Shigella flexneri (7), Klebsiella pneumonia (7), Enterobacter aerogenes (6), Enterobacter cloacae (5), Pseudomonas aeruginosa (5), Vibrio parahemolyticus (5), Aeromonas sobria (4), Citrobacter freundii (4), Citrobacter koseri (4) and Plesiomonas shigelloides (3). The detection of the common fecal coliforms (E. coli, K. pneumoniae and E. aerogenes) and Salmonella spp. in processed fish indicates the potential danger of passage of pathogenic bacteria and/or their poisons to humans via infected and/or contaminated fish products. Human infection by pathogenic fish bacteria and food poisoning is possible through contamination of fish product in fish production chain due to inadequate handling, poor hygiene and contact with contaminated water. Therefore, producers, consumers and all other stakeholders need to be cautious during handling, processing and consumption of fish harvested from the study lakes.
Topics: Animals; Bacteria; Carps; Cross-Sectional Studies; Escherichia coli; Fish Products; Gram-Negative Bacteria; Lakes; Water; Ethiopia
PubMed: 36517783
DOI: 10.1186/s12917-022-03508-w -
BMC Veterinary Research Apr 2022Bacterial infections are responsible of high economic losses in aquaculture. Mexican golden trout (Oncorhynchus chrysogaster) is a threatened native trout species that...
Bacterial and parasite co-infection in Mexican golden trout (Oncorhynchus chrysogaster) by Aeromonas bestiarum, Aeromonas sobria, Plesiomonas shigelloides and Ichthyobodo necator.
BACKGROUND
Bacterial infections are responsible of high economic losses in aquaculture. Mexican golden trout (Oncorhynchus chrysogaster) is a threatened native trout species that has been introduced in aquaculture both for species conservation and breeding for production and for which no studies of bacterial infections have been reported.
CASE PRESENTATION
Fish from juvenile stages of Mexican golden trout showed an infectious outbreak in a farm in co-culture with rainbow trout (Oncorhynchus mykiss), showing external puntiform red lesions around the mouth and caudal pedunculus resembling furuncles by Aeromonas spp. and causing an accumulated mortality of 91%. Isolation and molecular identification of bacteria from lesions and internal organs showed the presence of Aeromonas bestiarum, Aeromonas sobria, Plesiomonas shigelloides and Ichthyobodo necator isolated from a single individual. All bacterial isolates were resistant to amoxicillin-clavulanic acid and cefazoline. P. shigelloides was resistant to third generation β-lactamics.
CONCLUSIONS
This is the first report of coinfection by Aeromonas bestiarum, Aeromonas sobria, Plesiomonas shigelloides and Ichthyobodo necator in an individual of Mexican golden trout in co-culture with rainbow trout. Resistance to β-lactams suggests the acquisition of genetic determinants from water contamination by human- or livestock-associated activities.
Topics: Aeromonas; Animals; Coinfection; Fish Diseases; Gram-Negative Bacterial Infections; Necator; Oncorhynchus; Oncorhynchus mykiss; Parasites; Plesiomonas
PubMed: 35414073
DOI: 10.1186/s12917-022-03208-5