scholarly journals Molecular detection of glutamate dehydrogenase gene of Giardia lamblia isolated from food handlers in Erbil city

2021 ◽  
Vol 25 (1) ◽  
pp. 431-437
Author(s):  
Hataw Saber ◽  
Hawri Bakr
Keyword(s):  
Glutamate Dehydrogenase ◽  
Nested Pcr ◽  
Giardia Lamblia ◽  
Diarrheal Disease ◽  
Asymptomatic Carrier ◽  
Positive Sample ◽  
Pcr Analysis ◽  
Food Handler ◽  
Food Handlers ◽  
Dehydrogenase Gene

Background and objective: Giardia lamblia is the intestinal flagellated protozoan parasite that infects vertebrates, including humans. Giardiasis is the major diarrheal disease found worldwide. It can be symptomatic or may be an asymptomatic carrier that led to chronic disease. This study aimed to determine the proportion of giardiasis among food handlers and evaluate the correlation between two laboratory methods for identifying the Giardia lamblia. Methods: A total of 308 stool samples were collected from food handlers that annually attend the central laboratory in Erbil City. Wetmount microscopic examination was performed for the diagnosis of cysts and trophozoites of the Giardia parasite. Molecular analysis done for positive samples, DNA extraction performed using the QIAamp Fast DNA Stool Mini Kit (Qiagen Company, Germany). Nested PCR analysis was done targeting the Glutamate dehydrogenase gene using two sets of primers for amplification of 734bp fragment. Gel electrophoresis was performed for visualizing the amplified DNA by Ultraviolet light. Results: The mean age of food handler participants was 29 years. Most (93.8%) of the food handlers were males, and the majority (98.6%) of the participants did not have any signs and symptoms. Four (7.4%) microscopy positive sample participants were highly educated. There was no association between educational level and positive rate by microscopy (P = 0.066). The majority of participants did not receive treatments, particularly most of the microscopic positive samples. The food handlers did not take any antiparasitic treatments 9 (3.4%) (P = 0.676). From 11 (3.6%) microscopically positive samples, 10 (90.9%) Giardia lamblia gdh gene 734 bp fragments were amplified by nested PCR. Conclusion: Amplification of 734bp of gdh gene by the nested PCR is the most specific and sensitive method for identifying Giardia lamblia. Food handlers were important people to care about sanitation and preparing food, particularly for avoiding diseases transmitted by food. Keywords: Giardiasis; Genetic characterization; gdh gene; Nested PCR.

scholarly journals Transcriptional Analysis of the Glutamate Dehydrogenase Gene in the Primitive Eukaryote,Giardia lamblia

2000 ◽  
Vol 275 (15) ◽  
pp. 11432-11439 ◽  
Author(s):  
Janet Yee ◽  
Michael R. Mowatt ◽  
Patrick P. Dennis ◽  
Theodore E. Nash
Keyword(s):  
Glutamate Dehydrogenase ◽  
Giardia Lamblia ◽  
Transcriptional Analysis ◽  
Dehydrogenase Gene

scholarly journals A glutamate dehydrogenase gene sequence

1989 ◽  
Vol 17 (24) ◽  
pp. 10500-10500 ◽  
Author(s):  
J. Mark Cock ◽  
Robert R. Schmidt
Keyword(s):  
Glutamate Dehydrogenase ◽  
Gene Sequence ◽  
Dehydrogenase Gene

scholarly journals Spo0A Suppresses sin Locus Expression in Clostridioides difficile

mSphere ◽  
2020 ◽  
Vol 5 (6) ◽  
Author(s):  
Babita Adhikari Dhungel ◽  
Revathi Govind
Keyword(s):  
Diarrheal Disease ◽  
Toxin Production ◽  
The United States ◽  
Upstream Region ◽  
Pcr Analysis ◽  
Bacterial Cells ◽  
Master Regulator ◽  
Content Type ◽  
Clostridioides Difficile

ABSTRACT Clostridioides difficile is the leading cause of nosocomial infection and is the causative agent of antibiotic-associated diarrhea. The severity of the disease is directly associated with toxin production, and spores are responsible for the transmission and persistence of the organism. Previously, we characterized sin locus regulators SinR and SinR′ (we renamed it SinI), where SinR is the regulator of toxin production and sporulation. The SinI regulator acts as its antagonist. In Bacillus subtilis, Spo0A, the master regulator of sporulation, controls SinR by regulating the expression of its antagonist, sinI. However, the role of Spo0A in the expression of sinR and sinI in C. difficile had not yet been reported. In this study, we tested spo0A mutants in three different C. difficile strains, R20291, UK1, and JIR8094, to understand the role of Spo0A in sin locus expression. Western blot analysis revealed that spo0A mutants had increased SinR levels. Quantitative reverse transcription-PCR (qRT-PCR) analysis of its expression further supported these data. By carrying out genetic and biochemical assays, we show that Spo0A can bind to the upstream region of this locus to regulates its expression. This study provides vital information that Spo0A regulates the sin locus, which controls critical pathogenic traits such as sporulation, toxin production, and motility in C. difficile. IMPORTANCE Clostridioides difficile is the leading cause of antibiotic-associated diarrheal disease in the United States. During infection, C. difficile spores germinate, and the vegetative bacterial cells produce toxins that damage host tissue. In C. difficile, the sin locus is known to regulate both sporulation and toxin production. In this study, we show that Spo0A, the master regulator of sporulation, controls sin locus expression. Results from our study suggest that Spo0A directly regulates the expression of this locus by binding to its upstream DNA region. This observation adds new detail to the gene regulatory network that connects sporulation and toxin production in this pathogen.

Glucocorticoid upregulation of glutamate dehydrogenase gene expression in vitro in astrocytes

1996 ◽  
Vol 37 (1-2) ◽  
pp. 324-328 ◽  
Author(s):  
H. Hardin-Pouzet ◽  
P. Giraudon ◽  
M.F. Belin ◽  
M. Didier-Bazes
Keyword(s):  
Gene Expression ◽  
Glutamate Dehydrogenase ◽  
Dehydrogenase Gene

scholarly journals Microscopic And Molecular Identification of Anaplasma Ovis in Small Ruminants in Duhok Province in Kurdistan Region of Iraq

2020 ◽  
Vol 23 (2) ◽  
pp. 228-235
Author(s):  
Adnan Ahmed ◽  
Jassim M Abdo
Keyword(s):  
Microscopic Examination ◽  
Prevalence Rate ◽  
Small Ruminants ◽  
Surface Protein ◽  
Positive Sample ◽  
Pcr Analysis ◽  
Anaplasma Ovis ◽  
Blood Samples ◽  
Major Surface Protein ◽  
Major Surface

In last ten years, there has been a developing enthusiasm for microscopic organisms from the genus Anaplasma, particularly the species A. ovis. It is associated with the pathogenic action of these microscopic organisms in livestock. Anaplasma ovis is a tick-borne obligate intracellular rickettsial bacterium that causes anaplasmosis in domestic and wild small ruminants. The samples of the present study were collected from small ruminants from inside seven distinct regions (Akre, Simele, Zummar, Feshchapoor, Deraboon, Bajed Kandal,Karoda)of Duhok province, 389 (goats 75 and sheep 314) during the period of April and May 2018, blood sample were taken and thin smear was formed, after Giemsa’s staining the slide is observed under microscope. In this study used Giemsa stain for microscopic examination out of 389 animals 250 were found positive for Anaplasma ovis infection with a prevalence rate of 64.26 % and 139 of them were negative with a prevalence rate of 35.73 %. According to the species of animals, the highest prevalence of A. ovis infection in animals by using microscopic examination was 67.83 %, 213 positive sample from total 314 blood samples from sheep and lowest prevalence was 49.33 %, 37 positive sample from total 75 blood samples from goats. PCR analysis of 100 blood samples obtained from total 250 positive blood samples after DNA extraction and measure of concentration and purity we used 2 primers that target major surface protein 4 (MSP4) in A. ovis genomic DNA. The results of PCR test with major surface protein 4 primer was 83 samples positive from total 100 samples, According to the species of animals, the highest prevalence of A. ovis was 83.7 %, 72 positive sample from total 86 blood samples from sheep and lowest prevalence was 78.5 %, 11 positive sample from total 14 blood samples from goats.

scholarly journals Using RFLP-PCR Technique in Determining Genotypes of Giardia lamblia from Diarrhea Cases in Children in AL-Diwaniyah City, Iraq

2020 ◽  
Vol 1 (2) ◽  
pp. 29-34
Author(s):  
Hadi M. Hamza AL-Mayali ◽  
Lubna Abdul-Kadir AL-Ibrahim
Keyword(s):  
Rural Areas ◽  
Giardia Lamblia ◽  
Urban Areas ◽  
Age Groups ◽  
Intestinal Protozoa ◽  
Gender And Age ◽  
Dehydrogenase Gene ◽  
Urban And Rural Areas ◽  
Child Hospital ◽  
Genotype A

Introduction: Giardia lamblia is one of the most prevalent intestinal protozoa in the world, which affect children in both undeveloped and developing countries. This study aimed to determine genotypes of the Giardia lamblia using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP)-PCR techniques. Additionally, the relationship between genotype patterns and their geographical distribution, gender, and age was investigated. Methods: The current study included 926 samples of faeces of children suffering from diarrhoea, who visits the internal clinics at Teaching Hospital, and Child Hospital in AL- Diwaniyah City from November 2012 - Jun 2013. For age groups of 1-12 years exclusively. The samples were examined using a direct mount wet smear, The positive samples were preserved without adding preservatives at a temperature of -20°C until the DNA extraction for G. lamblia genotyping by using PCR and RFLP-PCR technique. Results: Giardia lamblia was detected in 2.15% (20/926) of samples from diarrhea cases in children by amplification of glutamate dehydrogenase gene (gdh) using two specific primers GDHiR and GDHiF. It was revealed that 7 samples belonged to genotype A (35%) and 13 samples belonged to genotype B (65%). All genotype A samples belonged to subgenotype AII (100%), while genotype B samples belonged to subgenotypes BIII (53.61 %) and BIV (47.38 %). Genotype A was detected in children of 1-6 years of age while B genotype was detected in all age groups. Both of the genotypes have been detected in both genders (male and female) and genotype B was found in both urban and rural areas; however, its prevalence was higher in rural areas than in urban areas (100% and 30%, respectively). Conclusion: There are two genotypes of Giardia lamblia, genotype A and genotype B, each of which has secondary genetic patterns which include AII, BIII, and BIV.

scholarly journals Investigation of Giardia intestinalis Genotypes among the Food Handlers of Qazvin, Iran

2019 ◽  
Author(s):  
Mojtaba SHAHNAZI ◽  
Farzaneh NAGHIZADEH ◽  
Elham HAJIALILO ◽  
Safar Ali ALIZADEH ◽  
Mehrzad SARAEI ◽  
...  
Keyword(s):  
Giardia Intestinalis ◽  
P Value ◽  
Giardia Cyst ◽  
Food Handlers ◽  
Dehydrogenase Gene ◽  
Stool Samples ◽  
Stool Specimens ◽  
The Impact ◽  
Concentration Methods

Background: We aimed to investigate the genotypes of Giardia intestinalis among the food handlers in Qazvin, Iran. Methods: Overall, 1530 stool specimens were collected from the food handlers who visited Shahid Bolandian Health Center, Qazvin, Iran during 2016. Specimens were evaluated by microscopic and concentration methods. Twenty specimens with appropriate number of giardia cysts were selected followed by DNA extraction. Determination of giardia genotypes was achieved through PCR and sequencing the glutamate dehydrogenase gene. The phylogenetic tree was drawn using the MEGA7 software. Finally, the data were analyzed statistically with a P-value<0.05 was considered as significant. Results: Twenty stool samples (1.3%) were positive for Giardia cyst. All positive specimens were obtained from male participants with abdominal cramp being their most common symptoms. The mean age for infected individuals was 32 yr. Molecular characterization was successfully performed for 17 isolates and two genotypes A (AII, 65%) and B (BIII, 35%) were identified. Conclusion: The most prevalent giardia genotypes among the food handlers in Qazvin were A (AII) and B (BIII) genotypes with A (AII) genotype as the dominant one in the region. Considering the direct association between the food handlers and public health as well as the impact of geographical and host conditions on dispersion and pathogenicity of various genotypes and their zoonotic aspects, further investigations are necessary.

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