scholarly journals Deteksi bakteri pembentuk amina biogenik pada ikan Scombridae secara multiplex PCR

2020 ◽  
Vol 23 (2) ◽  
pp. 359-371
Author(s):  
Rizsa Mustika Pertiwi ◽  
Mala Nurilmala ◽  
Asadatun Abdullah ◽  
Nurjanah ◽  
Roza Yusfiandayani ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Histidine Decarboxylase ◽  
Lysine Decarboxylase ◽  
Tyrosine Decarboxylase ◽  
Marine Broth

Amina biogenik merupakan komponen basa nitrogen yang terbentuk oleh dekarboksilasi asam amino. Amina biogenik yang sering terdeteksi pada produk perikanan di antaranya histamin, tiramin dan kadaverin. Gen pengkode dekarboksilasi asam amino tersebut yaitu histidine decarboxylase (hdc), tyrosine decarboxylase (tdc) serta lysine decarboxylase (ldc). Penelitian ini bertujuan untuk mendapatkan isolat DNA bakteri beserta konsentrasi dan kemurniannya, menyeleksi media pertumbuhan bakteri, mendeteksi gen hdc, tdc dan ldc menggunakan multiplex PCR serta identifikasi bakteri.. Sampel yang digunakan yaitu ikan tuna beku, tongkol beku, cakalang beku, tuna loin, pindang tongkol potong dan pindang tongkol bumbu kuning serta kontrol positif menggunakan ikan tuna yang disimpan pada suhu ruang selama 3 hari. Penelitian terdiri dari tahap pra-pengayaan bakteri pada media marine broth dan lactose broth, isolasi DNA sampel tanpa dan hasil pra-pengayaan, uji kemurnian dan konsentrasi isolat serta amplifikasi gen target hdc, tdc, ldc menggunakan multiplex PCR. Hasil yang diperoleh yaitu kualitas isolat DNA bakteri pada umumnya sudah murni, dengan konsentrasi isolat 15,75-157,84 ng/µL. Teknik multiplex PCR berhasil mendeteksi gen hdc, tdc dan ldc pada ikan scombridae. Kondisi optimum amplifikasi PCR yaitu annealing pada suhu 50°C selama 90 detik. Gen yang terdeteksi pada sampel tanpa pra-pengayaan yaitu tdc, sedangkan ldc dan hdc ditemukan pada sampel hasil pra-pengayaan media lactose broth dan marine broth. Bakteri pembentuk amina biogenik teridentifikasi sebagai Enterococcus fecalis, M. morganii, E. aerogenes, Citrobacter sp., Hafnia paralvei dan Obesumbacterium proteus. Amina biogenik merupakan komponen basa nitrogen yang terbentuk oleh dekarboksilasi asam amino. Amina biogenik yang sering terdeteksi pada produk perikanan di antaranya histamin, tiramin dan kadaverin. Gen pengkode dekarboksilasi asam amino tersebut yaitu histidine decarboxylase (hdc), tyrosine decarboxylase (tdc) serta lysine decarboxylase (ldc). Penelitian ini bertujuan untuk mendapatkan isolat DNA bakteri beserta konsentrasi dan kemurniannya, menyeleksi media pertumbuhan bakteri, mendeteksi gen hdc, tdc dan ldc menggunakan multiplex PCR serta identifikasi bakteri.. Sampel yang digunakan yaitu ikan tuna beku, tongkol beku, cakalang beku, tuna loin, pindang tongkol potong dan pindang tongkol bumbu kuning serta kontrol positif menggunakan ikan tuna yang disimpan pada suhu ruang selama 3 hari. Penelitian terdiri dari tahap pra-pengayaan bakteri pada media marine broth dan lactose broth, isolasi DNA sampel tanpa dan hasil pra-pengayaan, uji kemurnian dan konsentrasi isolat serta amplifikasi gen target hdc, tdc, ldc menggunakan multiplex PCR. Hasil yang diperoleh yaitu kualitas isolat DNA bakteri pada umumnya sudah murni, dengan konsentrasi isolat 15,75-157,84 ng/µL. Teknik multiplex PCR berhasil mendeteksi gen hdc, tdc dan ldc pada ikan scombridae. Kondisi optimum amplifikasi PCR yaitu annealing pada suhu 50°C selama 90 detik. Gen yang terdeteksi pada sampel tanpa pra-pengayaan yaitu tdc, sedangkan ldc dan hdc ditemukan pada sampel hasil pra-pengayaan media lactose broth dan marine broth. Bakteri pembentuk amina biogenik teridentifikasi sebagai Enterococcus fecalis, M. morganii, E. aerogenes, Citrobacter sp., Hafnia paralvei dan Obesumbacterium proteus.

Production of Tyrosine and Histidine Decarboxylase by Dairy-Related Bacteria

1977 ◽  
Vol 40 (4) ◽  
pp. 241-245 ◽  
Author(s):  
M. N. VOIGT ◽  
R. R. EITENMILLER
Keyword(s):  
Histidine Decarboxylase ◽  
Micrococcus Luteus ◽  
Coli Strain ◽  
Starter Cultures ◽  
Decarboxylase Activity ◽  
Tyrosine Decarboxylase ◽  
Streptococcus Lactis ◽  
Low Levels ◽  
Co2 Release ◽  
Manometric Technique

Manometric and radiometric procedures were used to determine the ability of 38 dairy-related bacteria and four commercial starter preparations to produce tyrosine and histidine decarboxylase. All of the cultures had slight ability to cause release of 14CO2 from carboxyl 14C-tyrosine and most released 14CO2 from labeled histidine; however, because of inherent errors of the assay in detecting low levels of specific decarboxylase activity, the CO2 release was not considered positive for specific decarboxylase activity unless the results were verified by the manometric technique. One strain of Streptococcus lactis, a Micrococcus luteus strain, and two Leuconostoc cremoris strains had active tyrosine decarboxylase systems. Only Clostridium perfringens and an Escherichia coli strain were found to produce histidine decarboxylase. None of the commercial starter cultures produced the enzymes. It was not determined whether tyramine or histamine was produced by the nonspecific decarboxylase activity.

scholarly journals Photobacterium angustum and Photobacterium kishitanii, Psychrotrophic High-Level Histamine-Producing Bacteria Indigenous to Tuna

2016 ◽  
Vol 82 (7) ◽  
pp. 2167-2176 ◽  
Author(s):  
K. Bjornsdottir-Butler ◽  
S. A. McCarthy ◽  
P. V. Dunlap ◽  
R. A. Benner
Keyword(s):  
United States ◽  
Histidine Decarboxylase ◽  
The United States ◽  
Optimum Growth ◽  
Marine Broth ◽  
Content Type ◽  
Histamine Production ◽  
Regulatory Limits ◽  
Fish Poisoning ◽  
High Level

ABSTRACTScombrotoxin fish poisoning (SFP) remains the main contributor of fish poisoning incidents in the United States, despite efforts to control its spread. Psychrotrophic histamine-producing bacteria (HPB) indigenous to scombrotoxin-forming fish may contribute to the incidence of SFP. We examined the gills, skin, and anal vents of yellowfin (n= 3), skipjack (n= 1), and albacore (n= 6) tuna for the presence of indigenous HPB. Thirteen HPB strains were isolated from the anal vent samples from albacore (n= 3) and yellowfin (n= 2) tuna. Four of these isolates were identified asPhotobacterium kishitaniiand nine isolates asPhotobacterium angustum; these isolates produced 560 to 603 and 1,582 to 2,338 ppm histamine in marine broth containing 1% histidine (25°C for 48 h), respectively. The optimum growth temperatures and salt concentrations were 26 to 27°C and 1% salt forP. kishitaniiand 30 to 32°C and 2% salt forP. angustumin Luria 70% seawater (LSW-70). The optimum activity of the HDC enzyme was at 15 to 30°C for both species. At 5°C,P. kishitaniiandP. angustumhad growth rates of 0.1 and 0.2 h−1, respectively, and the activities of histidine decarboxylase (HDC) enzymes were 71% and 63%, respectively. These results show that indigenous HPB in tuna are capable of growing at elevated and refrigeration temperatures. These findings demonstrate the need to examine the relationships between the rate of histamine production at refrigeration temperatures, seafood shelf life, and regulatory limits.

The effect of Tetraneura ulmi L. galling process on the activity of amino acid decarboxylases and the content of biogenic amines in Siberian elm tissues

2017 ◽  
Vol 108 (1) ◽  
pp. 69-76 ◽  
Author(s):  
K. Kmieć ◽  
C. Sempruch ◽  
G. Chrzanowski ◽  
P. Czerniewicz
Keyword(s):  
Amino Acid ◽  
Biogenic Amines ◽  
Ornithine Decarboxylase ◽  
Lysine Decarboxylase ◽  
Tyrosine Decarboxylase ◽  
Ulmus Pumila ◽  
Plant Organs ◽  
Key Enzymes ◽  
Initial Stage ◽  
Amine Content

AbstractTetraneura ulmi (L.), a member of Eriosomatinae subfamily, is one of the gall-forming aphids occurring on elms. Sap-sucking behaviour of founding mothers results in the formation of new plant organs. This study documents the changes in the content of plant biogenic amines (putrescine, cadaverine, spermidine, tryptamine, spermine and histamine) and key enzymes of their biosynthesis: lysine decarboxylase (LDC), tyrosine decarboxylase and ornithine decarboxylase (ODC) in galls and other parts of Siberian elm (Ulmus pumila L.) leaves during the galling process. The direction and intensity of these changes for particular amines and enzymes were dependent on the stage of gall development and part of the galling leaf. Generally, the amine content tended to increase in gall tissues during the 1st and 2nd period of the galling process and decreased in later phases. LDC and ODC activities were markedly enhanced, especially in gall tissues at the initial stage of the galling process.

Biogenic Amine Production by and Phylogenetic Analysis of 23 Photobacterium Species

2018 ◽  
Vol 81 (8) ◽  
pp. 1264-1274 ◽  
Author(s):  
K. BJORNSDOTTIR-BUTLER ◽  
A. ABRAHAM ◽  
A. HARPER ◽  
P. V. DUNLAP ◽  
R. A. BENNER
Keyword(s):  
Biogenic Amine ◽  
Bacterial Communities ◽  
Histidine Decarboxylase ◽  
Whole Genome ◽  
Lysine Decarboxylase ◽  
Conventional Pcr ◽  
Gene Characterization ◽  
Histamine Production ◽  
Current Guidelines

ABSTRACT Photobacterium species are members of the bacterial communities typically associated with scombrotoxin-forming fish. Reclassification and discovery of new Photobacterium species has caused confusion as to which species are capable of biogenic amine production. We analyzed histamine, cadaverine, and putrescine production by 104 Photobacterium strains representing 23 species. The presence of the genes for histidine decarboxylase (hdc), lysine decarboxylase (ldc), and ornithine decarboxylase (odc) was determined by real-time or conventional PCR and whole genome sequencing. Significant histamine production (>200 ppm) was detected in five Photobacterium species: P. angustum, P. aquimaris, P. kishitanii, P. damselae, and P. phosphoreum. The hdc gene was detected in all of these histamine-producing species except P. phosphoreum. Cadaverine was produced by eight Photobacterium species: P. angustum, P. aquimaris, P. damselae, P. iliopiscarium, P. kishitanii, P. leiognathi, P. mandapamensis, and P. phosphoreum. Putrescine was produced by six Photobacterium species: P. angustum, P. aquimaris, P. kishitanii, P. leiognathi, P. mandapamensis, and Photobacterium sp. Cadaverine production correlated closely with the presence of the ldc gene, but putrescine production did not correlate closely with the presence of the odc gene. Characterization of the biogenic amine production by Photobacterium species will allow identification of these marine bacteria and help ensure that current guidelines account for mitigation of these bacteria.

A multiplex PCR-SSCP test to genotype bovine β-casein alleles A1 , A2 , A3 , B, and C

1999 ◽  
Vol 30 (4) ◽  
pp. 322-323 ◽  
Author(s):  
A. Barroso ◽  
S. Dunner ◽  
J. Ca·ón
Keyword(s):  
Multiplex Pcr

Lysine decarboxylase assay with cucurbituril (cucurbit-7-uril)

2007 ◽  
Author(s):  
Andreas Hennig ◽  
Hüseyin Bakirci ◽  
Werner M. Nau
Keyword(s):  
Lysine Decarboxylase

Multiplex PCR zum Nachweis disseminierter Tumorzellen (DTC) im Blut von Patientinnen mit Mammakarzinom

2005 ◽  
Vol 127 (03) ◽  
Author(s):  
J Rom ◽  
A Schneeeweiss ◽  
V Zieglschmid ◽  
C Hollmann ◽  
O Böcher ◽  
...  
Keyword(s):  
Multiplex Pcr

Usefulness of a Multiplex PCR for Detection of Diarrheagenic Escherichia coli in a Diagnostic Microbiology Laboratory Setting

2016 ◽  
Vol 1 (2) ◽  
pp. 38-42 ◽  
Author(s):  
Khairun Nessa ◽  
Dilruba Ahmed ◽  
Johirul Islam ◽  
FM Lutful Kabir ◽  
M Anowar Hossain
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Clinical Laboratory ◽  
Proteinase K ◽  
Microbiology Laboratory ◽  
Pcr Assay ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Multiplex Pcr Assay ◽  
Diagnostic Microbiology

A multiplex PCR assay was evaluated for diagnosis of diarrheagenic Escherichia coli in stool samples of patients with diarrhoea submitted to a diagnostic microbiology laboratory. Two procedures of DNA template preparationproteinase K buffer method and the boiling method were evaluated to examine isolates of E. coli from 150 selected diarrhoeal cases. By proteinase K buffer method, 119 strains (79.3%) of E. coli were characterized to various categories by their genes that included 55.5% enteroaggregative E. coli (EAEC), 18.5% enterotoxigenic E. coli (ETEC), 1.7% enteropathogenic E. coli (EPEC), and 0.8% Shiga toxin-producing E. coli (STEC). Although boiling method was less time consuming (<24 hrs) and less costly (<8.0 US $/ per test) but was less efficient in typing E. coli compared to proteinase K method (41.3% vs. 79.3% ; p<0.001). The sensitivity and specificity of boiling method compared to proteinase K method was 48.7% and 87.1% while the positive and negative predictive value was 93.5% and 30.7%, respectively. The majority of pathogenic E. coli were detected in children (78.0%) under five years age with 53.3% under one year, and 68.7% of the children were male. Children under 5 years age were frequently infected with EAEC (71.6%) compared to ETEC (24.3%), EPEC (2.7%) and STEC (1.4%). The multiplex PCR assay could be effectively used as a rapid diagnostic tool for characterization of diarrheagenic E. coli using a single reaction tube in the clinical laboratory setting.Bangladesh J Med Microbiol 2007; 01 (02): 38-42

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