scholarly journals IDENTIFIKASI ZIGOSITAS IKAN LELE (Clarias gariepinus) TRANSGENIK F-2 YANG MEMBAWA GEN HORMON (PhGH) DENGAN MENGGUNAKAN METODE REALTIME-qPCR

2016 ◽  
Vol 11 (1) ◽  
pp. 39
Author(s):  
Huria Marnis ◽  
Bambang Iswanto ◽  
Romy Suprapto ◽  
Imron Imron ◽  
Raden Roro Sri Pudji Sinarni Dewi
Keyword(s):  
Real Time ◽  
Clarias Gariepinus ◽  
Fish Farming ◽  
Progeny Test ◽  
African Catfish ◽  
Mendelian Segregation ◽  
Qpcr Method ◽  
Pcr Method ◽  
Real Time Qpcr ◽  
Transgene Transmission

Produktivitas ikan budidaya dapat ditingkatkan melalui teknologi transgenesis. Populasi ikan lele transgenik cepat tumbuh telah dihasilkan dan karakter biologisnya telah diketahui. Namun informasi zigositas ikan lele transgenik perlu ditelaah lebih lanjut. Penelitian ini bertujuan untuk mengidentifikasi zigositas ikan lele transgenik F-2. Zigositas ikan lele transgenik diidentifikasi dengan menggunakan metode real-time qPCR (RT-qPCR) dan uji progeni. Identifikasi zigositas melalui uji progeni, dilakukan dengan mendeteksi transgen (PhGH) pada individu-individu F-3 hasil persilangan transgenik F-2 dengan non-transgenik. Hasil penelitian menunjukkan bahwa zigositas pada ikan lele transgenik F-2 dapat diidentifikasi dengan menggunakan metode RT-qPCR. Semua ikan transgenik F-2 adalah heterozigot, dengan nilai 2-Ct yang hampir sama tiap individu F-2, yaitu berkisar 0,80-0,99. Identifikasi zigositas dengan metode RT-qPCR menunjukkan hasil yang sama dengan uji progeni, semua transgenik F-2 tidak menghasilkan 100% anakan F-3 positif transgen. Pada uji progeni, transmisi transgen pada penelitian ini tidak mengikuti hukum segregasi Mendel, dengan kisaran sebesar 5%-40%.Fish farming productivity can be increased by transgenesis technology. On the previous study, transgenic African catfish population fast growing has been produced and its biological characters has been known. However information of transgenic zygosity of catfish should be examined. The aim of this study was to identify the zygosity of F-2 transgenic African catfish. The zygosity of F-2 transgenic was identified by real time-qPCR (RT-qPCR) method and progeny test. Further, identification of zygosity F-2 transgenic African catfish was confirmed by progeny test, while F-2 transgenic African catfish was mated with non-transgenic. Identification of zygosity F-2 transgenic was conducted by detection PhGH gene (transgene) in F-3 transgenic African catfish population. Transgene transmission was evaluated by PCR method. The result showed that the zygosity F-2 transgenic African catfish could be identified by RT-qPCR method. All F-2 transgenic African catfish were heterozygous, where as the 2-Ct value was almost same for all individual, which ranges from 0.80 to 0.99. The result of zygosity identification using RT-qPCR method was as same as that of progeny test. In the progeny test, transgene transmission in this study was non-Mendelian segregation, with ranges of 5%-40%.

scholarly journals Development of a Real-Time qPCR Method for Detection and Enumeration of Mycobacterium spp. in Surface Water

2010 ◽  
Vol 76 (21) ◽  
pp. 7348-7351 ◽  
Author(s):  
Nicolas Radomski ◽  
Françoise S. Lucas ◽  
Régis Moilleron ◽  
Emmanuelle Cambau ◽  
Sophie Haenn ◽  
...  
Keyword(s):  
Surface Water ◽  
16S Rrna ◽  
Real Time ◽  
Quantitative Pcr ◽  
Environmental Samples ◽  
Real Time Quantitative Pcr ◽  
Qpcr Method ◽  
Pcr Method ◽  
Real Time Qpcr ◽  
Mycobacterium Spp

ABSTRACT A real-time quantitative PCR method was developed for the detection and enumeration of Mycobacterium spp. from environmental samples and was compared to two other methods already described. The results showed that our method, targeting 16S rRNA, was more specific than the two previously published real-time quantitative PCR methods targeting another 16S rRNA locus and the hsp65 gene (100% versus 44% and 91%, respectively).

scholarly journals DETECTION, TRANSMISSION, AND EXPRESSION cDNA GROWTH HORMONE GENE (PhGH) OF STRIPPED CATFISH IN F-1 TRANSGENIC AFRICAN CATFISH

2014 ◽  
Vol 9 (2) ◽  
pp. 89
Author(s):  
Huria Marnis ◽  
Bambang Iswanto ◽  
Rommy Suprapto ◽  
Imron Imron ◽  
Raden Roro Sri Pudji Sinarni Dewi
Keyword(s):  
Growth Hormone ◽  
Real Time ◽  
Quantitative Pcr ◽  
African Catfish ◽  
Growth Hormone Gene ◽  
Real Time Quantitative Pcr ◽  
Qpcr Method ◽  
Gh Gene ◽  
Pcr Method ◽  
Transgenic Male

In previous study, the fast growth transgenic founder of African catfish was produced harboring a growth hormone (GH) gene construct containing a stripped catfish growth hormone (PhGH) cDNA. This study was conducted to investigate transgene (PhGH) transmission and expression in F-1 transgenic African catfish. The transgenic founders (female) were crossed with non-transgenic (male) to produce heterozygous F-1 progeny. PhGH gene was detected in the embryo, larvae, and seed of the transgenic F-1 using PCR method. Expression levels of transgene in embryo and larvae were analyzed using real-time quantitative PCR (qPCR) method. The transgene was detected in embryo, larvae and seed of F-1 transgenic African catfish. Founder could transmit PhGH gene to transgenic F-1 lines in ranged 36% to 48%. Expression level of Phgh gene in embryo was higher than that of the larvae; whereas in the embryo was 1.5 x 105 - 5.2 x 105 copies or 0.49-9.82 fold, while in the larvae was 1.1 x 105 - 2.5 x 105copies or 0.19-5.80 fold. 

scholarly journals Enzymes as Biomarkers of Environmental Stress in African Catfish (Clarias gariepinus) in Osun State, Nigeria

2017 ◽  
Vol 7 (14) ◽  
pp. 71-83
Author(s):  
Omolara Titilayo Aladesanmi ◽  
Femi Kayode Agboola ◽  
Rapheal Emuebe Okonji
Keyword(s):  
Heavy Metals ◽  
Heavy Metal ◽  
Anthropogenic Activities ◽  
Clarias Gariepinus ◽  
Fish Farming ◽  
Fish Tissue ◽  
Enzymatic Activities ◽  
African Catfish ◽  
Fish Ponds ◽  
Bioindicator Species

Background. Many natural aquatic bodies have been contaminated with heavy metals released from domestic, industrial and other anthropogenic activities. Fish are an important bioindicator species and play an important role in the monitoring of water pollution. Objectives. This study shows the effect of heavy metals on the distribution of glutathione S-transferases (GST), catalase, rhodanese and 3-mercaptopyruvate sulphur transferase (3-MST) isolated from the liver, gills, fins and muscle of Clarias gariepinus. Methods . Glutathione S-transferase, catalase, rhodanese and 3-mercaptopyruvate S-transferase enzymes were isolated from the liver and gills of fish by homogenization of each tissue (with specific buffers for each enzyme) and centrifugation. Serial dilutions of the crude enzymes were then assayed for residual enzymatic activities using standard enzyme assay protocol. Results. The results showed heavy metals in the liver and muscle of the investigated fish. This study indicated significant accumulation of heavy metals in the tissues/organ of the fish from Ilesha, Osogbo and Yakoyo fish ponds. These are three main towns in Osun State where the major occupation is fish farming. The relationship between enzymatic activities and heavy metal content in C gariepinus tissue showed positive and significant (p<0.05) correlations between lead (Pb) and GST as well as chromium (Cr) and GST. This implies that higher concentrations of Pb and Cr induced the expression of greater GST activity in the fish tissue. Conclusions. The study concluded that the pattern of response of GST, catalase, rhodanese and 3-MST activities in the various organs/tissues of C gariepinus to the heavy metals suggests that the excitation or inhibitions of their activities are organ specific. Further biochemical studies of fish tissues/organs are needed to characterize the enzymatic changes associated with heavy metal pollution. Competing Interests: The authors declare no competing financial interests

scholarly journals Investigation on the deletion and duplication of PMP22 gene in patients with Charcot-Marie-Tooth using Real-time PCR in Chaharmahal and Bakhtiari and Isfahan Provinces

2019 ◽  
Vol 21 (6) ◽  
pp. 254-257
Author(s):  
Masoumeh Pourhadi ◽  
Morteza Hashemzadeh-Chaleshtori ◽  
Mostafa Gholami ◽  
Mohammad-Saeed Jami
Keyword(s):  
Gene Duplication ◽  
Real Time ◽  
Internal Control ◽  
Real Time Pcr ◽  
Pmp22 Gene ◽  
Charcot Marie Tooth ◽  
Qpcr Method ◽  
Pcr Method ◽  
Hereditary Patterns ◽  
Prevalent Form

Background and aims: Charcot-Marie-Tooth (CMT) is a common sensory-motor polyneuropathy with a prevalence of 1/2500. It is divided into different subgroups and has various hereditary patterns. Among the different subgroups of CMT, type 1A is the most prevalent form of the disease, which is created due to the duplication of the PMP22 gene. In patients has a deletion in the PMP22 gene, the hereditary neuropathic disease is known to be liable to pressure. The aim of this study was to identify the patients affected by the disease with the new, simple, and fast qPCR method and to investigate the appropriateness of this method in evaluating these types of mutations. Methods: In this analytical-descriptive study (code:IR.SKUMS.REC.1394.152), gene duplication and deletion in the patients were studied using the Excel software. The blood samples of 15 families afflicted with CMT and 49 healthy individuals were collected in EDTA anticoagulant tubes and analyzed. DNA extraction and quantitative real-time PCR method were applied for the PMP22 gene as the target gene and the albumin gene as the internal control gene. Results: Two genes were compared in each patient, and it was found that 46% of the subjects had duplication in the PMP22 gene. Conclusion: The qPCR method is an easy and fast way to detect gene duplication and deletion in CMT patients. It does not require any statistical software and can be performed without needing for parental DNA. In addition, the results of this study are consistent with the results of various studies in some countries of the world where the highest levels of deletion and duplication in PMP22 gene are seen.

scholarly journals DISTRIBUTION AND EXPRESSION OF STRIPED CATFISH (Pangasionodon hypophtalmus) GROWTH HORMONE GENE (PhGH) IN THE ORGAN OF AFRICAN CATFISH (Clarias gariepinus) TRANSGENIC FOUNDER

2012 ◽  
Vol 7 (1) ◽  
pp. 11
Author(s):  
Huria Marnis ◽  
Raden Roro Sri Pudji Sinarni Dewi ◽  
Imron Imron ◽  
Bambang Iswanto
Keyword(s):  
Growth Hormone ◽  
Clarias Gariepinus ◽  
Stomach Muscle ◽  
African Catfish ◽  
Growth Hormone Gene ◽  
Caudal Fin ◽  
Expression Levels ◽  
Striped Catfish ◽  
Pcr Method ◽  
High Level

Faster growing African catfish can be produced by transgenesis. This study was conducted to investigate the distribution and expression of growth hormone gene (PhGH) in various organs of the transgenic African catfish (Clarias gariepinus) founder (F0). Transgene was detected using the PCR method in various organs, namely pituitary, brain, liver, heart, spleen, kidney, intestine, stomach, muscle, caudal fin, gill and eye. Transgene expression levels were analyzed using the method of reverse transcriptase-polymerase chain reaction (RT-PCR), -actin gene used as internal controls. The results showed that the PhGH was detected and expressed in all organs of the transgenic African catfish founder. The high level of PhGH expression was found in the liver, pituitary, intestine and brain; smaller amounts were detectable in muscle, spleen, kidneys, heart, and stomach, caudal fin, gill and eyes, range from 0.02-0.75 PhGH/-actin mRNA. The expression levels of PhGH had positive correlation with tissue and body size (P<0.05).

Antibiotic Resistance Profiling of Salmonella sp. isolated from African Catfish (Clarias gariepinus)

2021 ◽  
Vol 16 (3) ◽  
pp. 44-47
Author(s):  
P.T. Fowoyo
Keyword(s):  
Antibiotic Resistance ◽  
Pathogenic Bacteria ◽  
Clarias Gariepinus ◽  
Fish Farming ◽  
Selective Medium ◽  
Diffusion Method ◽  
African Catfish ◽  
Fresh Water Fish ◽  
Peptone Water ◽  
Amoxicillin Clavulanic Acid

The African catfish (Clarias gariepinus) is an important fresh water fish consumed by a large percentage of the populace globally and it may be contaminated by pathogenic bacteria such as Salmonella sp. In this study, a total of fifteen (15) samples of African catfish were collected from different markets in Lokoja, Nigeria. The Salmonella sp. were isolated from the catfish samples by pre-enrichment in peptone water and subsequent inoculation on selective medium namely brilliant-green agar (BGA), bismuth sulphite agar (BSA) and Salmonella-Shigella agar (SSA). The Salmonella isolates were tested for susceptibility to 10 different commercially available antibiotics using the disc diffusion method. A total of thirty-four Salmonella species was isolated. The percentage occurrence of Salmonella sp. in the catfishes examined was very high (80%). The incidence of Salmonella sp. in the intestine (86.7%) of the catfish was higher than for the gills (66.7%) and the skin (73.3%). Majority of the isolates were resistant to Amoxicillin, Sulfomethoxazole-trimethoprim, Amoxicillin-clavulanic acid and Streptomycin. This study therefore demonstrated the occurrence of Salmonella species in African catfish with some exhibiting antibiotic resistance. Thus, there is a potential risk of transmission of drug resistant Salmonella species to man when contaminated catfish is consumed. The use of antibiotics in fish farming should be regulated so as to decrease antibiotic residues in fish.

SYBR Green real-time qPCR method: Diagnose drowning more rapidly and accurately

2021 ◽  
Vol 321 ◽  
pp. 110720
Author(s):  
Zhonghao Yu ◽  
Quyi Xu ◽  
Cheng Xiao ◽  
Huan Li ◽  
Weibin Wu ◽  
...  
Keyword(s):  
Real Time ◽  
Sybr Green ◽  
Qpcr Method ◽  
Real Time Qpcr

scholarly journals Effect Of Commercial Biofertilizer Product Of Seaweed (Euchema Cottonii) Towards The Growth Of Pakcoy Mustard (Brassica Rapa L) and Dumbo Catfish (Clarias Gariepinus) in Aquaponic System

2020 ◽  
Vol 9 (2) ◽  
pp. 86
Author(s):  
Oktaviana Elsa Dewi ◽  
Sapto Andriyono ◽  
Mochammad Amin Alamsjah
Keyword(s):  
Survival Rate ◽  
Clarias Gariepinus ◽  
Fish Farming ◽  
African Catfish ◽  
Optimal Dosage ◽  
Vegetable Production ◽  
Randomized Design ◽  
Growing Medium ◽  
Completely Randomized Design ◽  
Biomass Plant

Aquaponics is a technology that combines the cultivation of vegetable production and fish farming. The aquaponics technology also to fertilization to give nutrients to the plant to support of survival. In facts, fertilization done to plants is located on land. Fertilizing the plants that live in the waters are still very rare, because the waters as a growing medium that is considered to always provide sufficient nutrients for plant growth. In this research, using biofertilizer Euchema cottonii of seaweed. The advantages of biofertilizer seaweed is to have the content of macro and micro nutrients are complete. It also contains substances hyper Grow (PGR) plant (auxin, cytokinin, gibberellin, abisat acid and ethylene).In this research to determine the effect of biofertilizer products of seaweed (E. cottonii) commercial on the growth of dumbo catfish (C. gariepinus) and pakcoy mustard (B. rapa L) on aquaponics system. The design of experiments in this research using a completely randomized design (CRD) with 5 treatments and 4 replications. The treatment used is fertilizer "AB mix®" 1% (A), without fertilizer (B) and the provision of products biofertilizer seaweed (E. cottonii) commercial 0,16% (C), 0,18% (D) and 0,20% (E). The main parameters measured were heavy growth of biomass, plant height pakcoy mustard (B. rapa L), growth SGR, length and survival rate of fish of dumbo catfish (C. gariepinus).The results showed that the dose of different commercial product biofertilizer seaweed (E. cottonii) gives a significantly different effect on the height growth of pakcoy mustard (B. rapa L) and the survival rate of dumbo catfish (C. gariepinus). The optimal dosage of commercial seaweed (E. cottonii) biofertilizer product in increasing the growth of pakcoy mustard (B. rapa L.) and African catfish (C. gariepinus) in the aquaponic system, namely treatment E at a dose of 0.20%.

scholarly journals Antibacterial Activity of Isolates from Digestive Tract of Catfish (Clarias gariepinus) As Probiotic Candidates At PT. Biotech Cipta Kreasi

2021 ◽  
Vol 3 (1) ◽  
pp. 138-149
Author(s):  
Toto Apriliyanto ◽  
Nosa Septiana Anindita ◽  
Wisnu Adhi Susila
Keyword(s):  
Digestive Tract ◽  
Aeromonas Hydrophila ◽  
Low Cost ◽  
Clarias Gariepinus ◽  
Fish Farming ◽  
African Catfish ◽  
Bacillus Sp ◽  
High Level ◽  
Control Management ◽  
Probiotic Activity

African catfish (Clarias gariepinus) is a popular fish in the community because it provides a high level of protein at a low cost. The motile aeromonas septicemia (MAS) disease produced by Aeromonas hydrophila (A. hydrophila) infection is one of the things that create difficulties in the cultivation of this fish. Synthetic antibiotics are commonly used in MAS disease control management efforts in fish farming, which have have an influence not only on fish but also on consumers. The application of probiotic Bacillus sp isolated from the digestive tract of catfish as a means of preventing the sickness is one option. Enzymatic tests (lipolytic, amylolytic, cellulolytic, proteolytic activity) and antagonist testing were used to assess the probiotic activity of the Bacillus sp isolates.

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