scholarly journals Genetic characterization of Lactococcus garvieae isolated from farmed rainbow trout by random amplified polymorphic DNA-PCR in Iran

2020 ◽  
Vol 71 (1) ◽  
pp. 2023
Author(s):  
F. FADAEIFARD ◽  
M. RABIEI ◽  
M. F. SHARIFPOUR
Keyword(s):  
Rainbow Trout ◽  
Bacterial Infections ◽  
Random Amplified Polymorphic Dna ◽  
Farmed Fish ◽  
Biochemical Tests ◽  
Bacterial Cultures ◽  
Lactococcus Garvieae ◽  
Strain Pattern ◽  
Rapd Pcr ◽  
Pcr Method

Lactococcosis is one of the main bacterial infections of fish around the world. Lactococcus garvieae has been a major cause of rainbow trout losses in freshwater farming. This study aimed to genotype and determine the variability of L. garvieae isolated from infected farmed rainbow trout in Iran by the RAPD-PCR method. Bacterial samples were collected from 12 farms located in the western part of Iran and suspected to carry Lactococcus infection. Two hundred bacterial cultures containing cocci shaped bacteria were cultured in Trypticase soy agar (TSA) and blood agar mediums. All bacterial cultures were tested by conventional microbiological and biochemical tests, and PCR assay to identify L. garvieae by 16S rDNA genes. The RAPD-PCR method was used to determine the genetic pattern of all isolates. The sample strain pattern of the isolates was analyzed in the NTSYS program. According to a similarity coefficient index of 70%, all L. garvieae isolates were separated into two groups with four RAPD profile types. The highest and the lowest genetic pairwise similarity among the isolates were 98% and 54%, respectively. The results of the present study revealed that RAPD-PCR is an applicable method to describe the genetic diversity of different strains of L .garvieae among farmed fish.

scholarly journals Study of pathogenicity and severity of Lactococcus garvieae isolated from rainbow trout (Oncorhynchus mykiss) farms in Kohkilooieh and Boyerahmad province

2019 ◽  
Vol 22 (1) ◽  
Author(s):  
Esmaeil Karami ◽  
Mojtaba Alishahi ◽  
Taravat Molayemraftar ◽  
Masoud Ghorbanpour ◽  
Mohammad Reza Tabandeh ◽  
...  
Keyword(s):  
Rainbow Trout ◽  
Systemic Infection ◽  
Fish Farms ◽  
Pcr Assay ◽  
Biochemical Tests ◽  
High Severity ◽  
Lactococcus Garvieae ◽  
Specific Pcr ◽  
Rainbow Trout Oncorhynchus Mykiss ◽  
Trout Farms

Abstract Background Lactococcus garvieae is one of the most important risk factors in the rainbow trout culture. Therefore, the purpose of this study was to identify and detect strains isolated from rainbow trout suspected of having Lactococcus garvieae using biochemical characteristics and PCR and determination of the degree of severity of isolated strains. Methods In this study, the cause of lactococcosis in selected rainbow trout farms in Kohkilooieh and Boyerahmad province was assayed. Gram-positive and catalase-negative bacterial isolates were first obtained from selected trout fish farms using conventional biochemical tests and PCR assay. The 10-day LD50 method (concentration causing 50% mortality in 10 days) was used to determine the severity of the isolated bacteria. Results One bacterial isolate was detected from all sampled fish which confirmed as Lactococcus garvieae using a specific PCR assay based on the 16S rDNA gene by producing a single band of 1107 bp. Analysis of the rate of mortality showed that the 10-day LD50 was 4.6 × 105 CFU/fish. The results of this study showed that isolated bacteria had high severity for rainbow trout. The presence of bacteria in internal organs of suspected fish showed a severe systemic infection in challenged fish. Antibiogram assay also indicated that the isolated Lactococcus garvieae were resistant to some mostly used antibiotics in rainbow trout. Conclusions According to current research, it can be concluded that the condition of lactococcosis in the studied area is not suitable, and despite the presence of disease, there is no proper action to control and prevent the disease. Unfortunately, isolated bacteria from the studied area have a very high severity compared to bacteria isolated from other regions of the country or other countries. Therefore, further investigation is needed to determine the cause of this difference and possibly in the design of the vaccine.

scholarly journals Aeromonas veronii and Plesiomonas shigelloides (Gammaproteobacteria) isolated from Glossogobius aureus (Gobiidae) in Lake Sampaloc, Laguna, Philippines

2020 ◽  
Vol 14 (1) ◽  
Keyword(s):  
Bacterial Infections ◽  
Management Practices ◽  
16S Rrna Gene Sequencing ◽  
Gut Bacteria ◽  
Inland Waters ◽  
Rrna Gene ◽  
Farmed Fish ◽  
Biochemical Tests ◽  
Plesiomonas Shigelloides ◽  
Aeromonas Veronii

Bacterial infections are high-risk factors in fisheries, with reports of high mortality among diseased fish stocks posing a threat to both capture and aquaculture fisheries in inland waters. Diseases-causing bacteria in fishes may lead to decreased yield and economic loss to fishers, whose livelihood primarily depends on landed catch. Lake fisheries are most affected by such disruptive changes because of limitations in water turnover aggravated by wastewater inputs. In this study, we isolated and characterized gut bacteria from landed catch of the gobiid Glossogobius aureus from Lake Sampaloc, a small but commercially important aquaculture area in Luzon. Isolated axenic gut bacteria were identified through Gram stain reaction, microscopy, API biochemical tests, and 16s rRNA gene sequencing. From these, we identified two species with known fish pathogenicity, namely Aeromonas veronii and Plesiomonas shigelloides which are known to thrive in disrupted and nutrient-rich habitats and cause visible damage to fish health. Interestingly, our samples have shown no such visible signs of the disease. It is therefore important for future researches to determine what conservation and management practices in small inland waters like lakes will limit potential environmental stressors that may trigger susceptibility of both capture and farmed fish species to infection. Ultimately, rehabilitation of inland water aquaculture areas such as Lake Sampaloc is essential not only to fish conservation but also to public health and local food security.

scholarly journals Genotyping Of Lactococcus garvieae Isolated From Rainbow Trout By RAPD-PCR Using M13 And P5 Primers In Turkey

2016 ◽  
Vol 3 ◽  
Author(s):  
DUMAN Muhammed ◽  
ALTUN Soner ◽  
SATICIOĞLU Izzet ◽  
BÜYÜKEKİZ Ayşe
Keyword(s):  
Rainbow Trout ◽  
Lactococcus Garvieae ◽  
Rapd Pcr

scholarly journals Molecular analysis and dimorphism of azole-susceptible and resistant Candida albicans isolates

2011 ◽  
Vol 44 (6) ◽  
pp. 740-744 ◽  
Author(s):  
Carolina Rodrigues Costa ◽  
Lúcia Kioko Hasimoto e Souza ◽  
Fábio Silvestre Ataídes ◽  
Pedro Henrique Ferri ◽  
Maysa Paula da Costa ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Antifungal Agents ◽  
Random Amplified Polymorphic Dna ◽  
Induction Medium ◽  
Genotypic Characterization ◽  
Rapd Pcr ◽  
Group A ◽  
Pcr Method ◽  
Hyphal Formation ◽  
Group B

INTRODUCTION: Candida albicans is responsible for superficial or systemic infections known as candidiasis, which may be found in infected tissue as unicellular budding yeasts, hyphae, or pseudohyphae. In this study, the effects of both fluconazole and itraconazole antifungal agents on the hyphal formation and genotypic characterization of C. albicans isolates classified as either susceptible or resistant were investigated. METHODS: The hyphal production of five C. albicans isolates under the action of antifungal agents was investigated by culturing yeast on growth medium and on hyphal induction medium. The genotypic characterization was carried out for 13 isolates of C. albicans using the random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) method. RESULTS: The dimorphism analysis showed that the hyphal formation was higher in resistant than in the susceptible isolates to both azoles. The RAPD-PCR method identified the formation of two different groups. In group A, four resistant and two susceptible isolates were clustered, and in group B, one resistant and six susceptible isolates were clustered. CONCLUSIONS: Considering that hyphal formation was higher in resistant isolates in the presence of azole drugs, we confirmed that the hyphal production is closely related to susceptibility to azoles. These drugs may affect the morphogenesis of C. albicans depending on their susceptibility to these drugs. In relation to RAPD-PCR, most resistant isolates classified in group A and susceptible isolates in group B demonstrated that this method presented a similar standard between the two groups, suggesting that by this technique, a strong correlation between genotypes and fluconazole-resistant samples may be found.

scholarly journals First Identification and Characterization of Lactococcus garvieae Isolated from Rainbow Trout (Oncorhynchus mykiss) Cultured in Mexico

Animals ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1609
Author(s):  
Cesar Ortega ◽  
Rute Irgang ◽  
Benjamín Valladares-Carranza ◽  
Constanza Collarte ◽  
Ruben Avendaño-Herrera
Keyword(s):  
Rainbow Trout ◽  
Oncorhynchus Mykiss ◽  
Consensus Sequence ◽  
Fish Farming ◽  
16S Rrna Gene Sequencing ◽  
Surveillance Program ◽  
Rrna Gene ◽  
Biochemical Tests ◽  
Lactococcus Garvieae ◽  
Rainbow Trout Oncorhynchus Mykiss

Lactococcosis is a hyperacute hemorrhagic septicemia disease caused by Lactococcus garvieae, which is an emerging pathogen in global fish farming. Between 2016 and 2018, rainbow trout (Oncorhynchus mykiss) from five farms that presented outbreaks were sampled as part of a Mexican surveillance program for the detection of fish diseases. Fourteen L. garvieae isolates were recovered from sampled fish, as confirmed by biochemical tests, 16S rRNA gene sequencing, and clinical and histological insights. The biochemical and protein profiles of the isolates obtained were homogeneous. Repetitive extragenic palindromic—(REP)—and enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR) analyses established weak genetic heterogeneity. Rainbow trout challenged with two of the isolates used at different bacterial concentrations (10−2 and 10−4 CFU/mL) showed melanosis, and hemorrhages were observed in the fins, liver, kidney, and spleen. Isolates were obtained from all of the organs sampled, including from surviving fish, as either pure or mixed cultures. The present study is the first to confirm the presence of L. garvieae as the agent of severe lactococcosis outbreaks in the two primary Mexican states for trout farming.

scholarly journals Determination of Genetic Diversity of Four Guppy Strains Using the Random Amplified Polymorphic DNA-PCR

2021 ◽  
pp. 1-8
Author(s):  
Riva Hafidah ◽  
Ayi Yustiati ◽  
Yuniar Mulyani ◽  
Ibnu Bangkit Bioshina Suryadi
Keyword(s):  
Genetic Diversity ◽  
Random Amplified Polymorphic Dna ◽  
Descriptive Analysis ◽  
Similarity Index ◽  
Quantitative Descriptive Analysis ◽  
Rapd Pcr ◽  
Result Show ◽  
Pcr Method ◽  
Quantitative Descriptive

This research aims to determine genetic diversity of four strains guppy, respectively are japan blue double sword (JBD), japan blue tiger double sword (JBTD), blue moscow (BM), and panda guppy (PG) with RAPD-PCR method. The obtained genetic diversity data is used as guide reference for hybridization between four strains. The research was conducted in September 2020 to April 2021 with explorative methods and in qualitative and quantitative descriptive analysis. The research was carried out in biotechnology Laboratory, Fishery and Marine Sciences Faculty and Central Laboratory,Padjadjaran University, Indonesia.Strains of JBD, JBTD, BM obtained fromCilengkrangSubdistrict, Bandung and PG strain obtained from Parung market, Bogor. Primary OPA-03 (AGTCAGCCAC) is used for standard parameters to interpret genetic diversity among four strains of guppy. Based on results, amplification with OPA-03 primary visualize 25 bands that include five polymorphic bands and 20 monomorphic bands. The phylogenetic tree result show that there are two relationship groups. The first group are JBD, JBTD, and BM with similarity index in the range of 80-89%The first group consist two sub groups of relationship. The first sub group are JBD and JBTD with similarity index of 89%. The second sub group is BM with similarity index of 80%. The second group isPG with similarity index of 65.5%.

scholarly journals Antibacterial Effects of Octenidine Dihydrochloride, and Benzalkanium Chloride on Acinetobacter Baumannii Strains Isolated from Clinical Samples and Determination of Genetic Diversity of Isolates by RAPD-PCR Method

2021 ◽  
Author(s):  
Azar Dokht Khosravi ◽  
Effat Abasi Montazeri ◽  
Seyedeh Roya Maki
Keyword(s):  
Genetic Diversity ◽  
Acinetobacter Baumannii ◽  
Clinical Samples ◽  
Biochemical Tests ◽  
Typing Method ◽  
Conventional Culture ◽  
Rapd Pcr ◽  
Octenidine Dihydrochloride ◽  
Pcr Method

Abstract Due to the emergence of antibiotic resistance in Acinetobacter baumannii which is one of the important causes of nosocomial infections, many problems have been raised in the successful treatment of patients with the subsequent mortality. So, the present study was performed to evaluate the antibacterial effect of Actinidine dehydrochloride, Actinisept, and Benzalkanium chloride against Acinetobacter baumannii strains isolated from clinical samples and to determine the genetic diversity of strains by RAPD-PCR. A total of 119 non-duplicate, suspected Acinetobacter baumannii isolates were collected and confirmed by conventional culture and biochemical tests and PCR technique. Susceptibility of the isolates to antibiotics was evaluated by standard Antibiotic susceptibility testing (AST). For antiseptics Octenidine dihydrochloride (OCT), Actinisept, and Benzalkonium chloride (BZK), Minimal inhibitory concentration (MIC) was assessed. The prevalence of Qac E and Qac delta E genes related to antiseptics was estimated by PCR. Finally, genetic diversity of strains was determined by RAPD-PCR. All 119 suspected isolates were confirmed as Acinetobacter baumannii using conventional tests and PCR. The isolates were mostly originated from blood samples. In AST, the lowest resistance was seen for ciprofloxacin and gentamicin. The MIC values were reported as OCT (15.26 µg) and BZK (640 µg). The antiseptic genes of qacE and qac ΔE1 were found to be present in 56 (47.05%) and 59 (49.57%) of isolates respectively. RAPD typing method revealed great diversity among A. baumannii isolates, with 37 clusters in isolates from ICU, of which 32 isolates were single and 5 were multiple. In conclusion, considering the increase of resistance to antiseptics, it is of importance to monitor the susceptibility of A. baumannii to antiseptics and to promote antiseptic stewardship in hospitals. Furthermore, in this study great diversity among A. baumannii was observed making it difficult to properly carry out infection control policies. analysis of RAPD-PCR typing results, and we found 37 clusters, among them 32 isolates were single and 5 were multiple. So, the method generated 37 RAPD type which shows great diversity among 57 out of 62 A. baumannii isolates at 80% cutoff.

scholarly journals Genetic structure and diversity among three Greek sheep breeds using Random Amplified Polymorphic DNA-PCR

2017 ◽  
Vol 62 (4) ◽  
pp. 301 ◽  
Author(s):  
I. MASTRANESTASIS (Ι. ΜΑΣΤΡΑΝΕΣΤΑΣΗΣ) ◽  
Ch. LIGDA (Χρ. ΛΙΓΔΑ) ◽  
K. THEODOROU (Κ. ΘΕΟΔΩΡΟΥ) ◽  
L. V. EKATERINIADOU (Λ.Β. ΑΙΚΑΤΕΡΙΝΙΑΔΟΥ)
Keyword(s):  
Genetic Structure ◽  
Genetic Differentiation ◽  
Phylogenetic Relationships ◽  
Random Amplified Polymorphic Dna ◽  
Genetic Distances ◽  
Genetic Identity ◽  
Rapd Pcr ◽  
Pcr Method ◽  
Genetic Structure And Diversity

Genetic structure and diversity of 120 animals from three Greek local breeds were investigated by Random Amplified Polymorphic DNA (RAPD) - PCR method. Sheep samples originated from the Lesvos, Chios and Karagouniko breeds were treated with 11 random primers to estimate their genetic diversity and phylogenetic relationships. Our analysis comprised two levels of the breeds' genetic structure: i) the genetic differentiation among the three breeds and ii) the genetic differentiation among the flocks within each breed. This combined approach gave two main findings: i) the study of genetic distances and identity revealed that the Karagouniko sheep breed is genetically distinct from Chios (GD=0.1979) and Lesvos (GD=0.1691) breeds, while ii) the Chios and Lesvos breeds are genetically similar (GI=0.9631); half of the flocks of Lesvos have a relatively closer relationship with those of Chios than with the other Lesvos flocks. This is the first study that reports the close genetic relationship between the Chios and Lesvos breeds and gives strong evidence to hypotheses about their related origin. Furthermore, the study of polymorphic loci revealed particular indicators located in Karagouniko breed, as definitional datum of genetic identity or as a fingerprint of breed. Therefore, RAPD-DNA methods can be an efficient tool for the determination of phylogenetic relationships and genetic identity among the bloodstock breed of sheep.

scholarly journals Conventional and Molecular Typing of Salmonella enterica serotype Typhi Locally Isolated In Baghdad

2012 ◽  
Vol 9 (4) ◽  
pp. 632-639
Author(s):  
Baghdad Science Journal
Keyword(s):  
Molecular Typing ◽  
Salmonella Enterica ◽  
Random Amplified Polymorphic Dna ◽  
Discriminatory Power ◽  
Universal Primers ◽  
Pcr Analysis ◽  
Biochemical Tests ◽  
Promising Alternative ◽  
Rapd Pcr ◽  
Typing Methods

Phenotypic And genotypic characteristics of Salmonella enterica serotype Typhi have been determined for 29 isolates, from Baghdad in 2007. Conventional typing methods were performed by biochemical tests, and antimicrobial susceptibility test. Molecular typing performed by analysis plasmid DNA beside using the Random Amplified Polymorphic DNA (RAPD-PCR). For the latter, two universal primers that have selected for the high discriminatory power were used for RAPD analysis. All isolates were belong one biotype according to the differention by their ability to decarboxylat lysine, 29(100%) were lysine (+). All the isolates were susceptible to the Antibiotics used. However, all the strains free of plasmids. RAPD was capable of grouping the strains in 6 genotypic patterns using primer 784, in 4 genotypic patterns using primer 787. Conventional phenotypic typing methods, as well as the DNA plasmid analysis, presented non significant discriminatory power; however, RAPD-PCR analysis showed discriminatory power, reproducibility, easy interpretation and can be considered as a promising alternative typing method for S. Typhi.

scholarly journals Conventional and molecular typing of Salmonella typhi strains from Brazil

2002 ◽  
Vol 44 (6) ◽  
pp. 315-319 ◽  
Author(s):  
Bianca R. QUINTAES ◽  
Nilma C. LEAL ◽  
Eliane M. F. REIS ◽  
Érica L. FONSECA ◽  
Ernesto HOFER
Keyword(s):  
Molecular Typing ◽  
Random Amplified Polymorphic Dna ◽  
Salmonella Typhi ◽  
Discriminatory Power ◽  
Pcr Analysis ◽  
Biochemical Tests ◽  
Promising Alternative ◽  
Phage Types ◽  
Rapd Pcr ◽  
Typing Methods

Phenotypic and genotypic characteristics of Salmonella Typhi were studied in 30 strains, isolated in different years, from some areas in Brazil. Conventional typing methods were performed by biochemical tests, Vi phage-typing scheme, and antimicrobial susceptibility test. Molecular typing methods were performed by analysis of plasmid DNA and by random amplified polymorphic DNA (RAPD-PCR). For the latter, an optimization step was performed to ensure the reproducibility of the process in genetic characterization of S. Typhi. The predominance of 76.7% of biotype I (xylose +, arabinose -) was noticed in all studied areas. Three phage types were recognized, with prominence for the phage types A (73.3%) and I+IV (23.3%). All the strains were susceptible to the drugs used. However, 36.7% of the strains contained plasmids, with predominance of the 105 Kb plasmid. RAPD was capable of grouping the strains in 8 genotypic patterns using primer 784, in 6, using primer 787 and in 7, using primer 797. Conventional phenotypic typing methods, as well as the DNA plasmid analysis, presented nonsignificant discriminatory power; however, RAPD-PCR analysis showed discriminatory power, reproducibility, easy interpretation and performance, being considered as a promising alternative typing method for S. Typhi.

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