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IJCRR - 7(11), June, 2015

Pages: 13-19

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Molecular detection and discrimination of three poultry Eimeria species in Kerbala and Babylon provinces, Iraq

Author: Dhamiaa Make Hamza, Hadi Rasol Hasan Al-Massodi, Zuhair Muhammad Ali Jeddoa

Category: Healthcare

Abstract:Coccidiosis is an intestinal disease of chickens in which the Causes include a various species of protozoan parasites that located within the genus Eimeria. Diagnosis and genetic characterization of different species of Eimeria are central dogma to the preventative, resistance, and control of coccidiosis. The aim of the present study was to identified different avian Eimeria species that considered the causal agents of avian coccidiosis in each of the Kerbala and Babylon provinces, Iraq. The present study observations were revealed increasing and decreasing the percentages of infection with coccidiosis during different months of a year and that due to the large change in temperature and humidity in Iraq, so the results showed that the highest percentage of infection in both of Kerbala and Babylon provinces were in December (23.12%) and the lowest percentage in July (1.87%). A total of 200 samples of feces and intestines were collected from chickens that suspected to infection with Eimeria during the period from August 2013 to July 2014 and the results revealed that 160 samples were positive depending on the macroscopic and microscopic examination which in turn have undergone the second stage of the examination for the purpose of qualitative accurate diagnosis of the species of Eimeria that causing coccidia in chickens using of molecular methods for the target region ITS1 nucleic acid DNA encoded ribosomal RNA , these techniques depend on qRT- PCR technique ,Which diagnosed three species of Eimeria includes E. tenella which formed the largest proportion of the total positive samples (66.8%) then the species E. necatrix which accounted for (30%) and finally the species E. maxima which form (3.13%).Therefore , Using of the molecular methods such as REAL-TIME PCR which characterized by highly accuracy, but these methods expensive economically compared
to routine methods.

Keywords: Avian Coccidiosis, Eimeria, Oocysts, RT-PCR, Iraq

Full Text:

INTRODUCTION

Avian coccidiosis is one of the most important diseases affecting the intensive poultry industry worldwide. Coccidia are almost usually found wherever chickens are raised and it is exceedingly rare to find a commercial chicken flock not affected. The infection causes tissue damage in the intestinal tract leading to interruption in digestive processes, blood loss, and increased susceptibility to other diseases, subclinical enteric infection, and sub acute mortality, the disease is treated by used of anticoccidial drugs that added in the feed or vaccination (1, 2, 3, 4). There are nine commonly recognized species of chicken coccidia; Eimeria acervulina, E. brunetti, E. maxima , E. mitis, E. necatrix, E. praecox , E.mevati , E. hagani and E. tenella, each Eimeria species develops in a particular location within the chicken digestive tract with some interfere seen between these species(5). The specific identification of Eimeria species and strains is important for diagnosis and domination, as well as for epidemiology and biological studies of the population (6).Depended on the routine tests, the Eimeria species have been identified by morphology and/or morphometry of their sporocysts and oocysts as well as their modes of development, and assessing the site and extent of the pathological lesions in the intestine of chicken (7, 8). However, these ways are costly, time-consuming, require skilled personnel and can be unreliable under the circumstances of mixed field infections, particularly when the overlap in biological and morphological characteristics makes the unambiguously identification and differentiation of Eimeria species impossible (9, 10). Molecular techniques have some advantages over traditional methods in that they rely only on the genomic sequence of the Eimeria species, several techniques based on the polymerase chain reaction using primers that specifically targeting different regions of the Eimeria genome have been described (5, 11, 12, 13, 14, 15). For molecular detection of Eimeria species in chickens, the DNA sequence of the first and second internal transcribed spacers (ITS-1 and ITS-2) of the nuclear DNA, which separate the ribosomal genes, is used most frequently, besides its heterogeneity in both sequence length and base composition of the ITS sequence, the rDNA is a member of a multiple copy gene family and provides large numbers of potential PCR targets (16, 17, 18, 19, 20). Nevertheless, the practical implementation of these techniques in tradition diagnosis and epidemiological studies has been limited (21). The number of studies related to the presence of chicken Eimeria species in Iraq are limited. These studies have been conducted in few locations and species identification was depended on the oocysts morphology, necropsy and/or histopathological findings. And as a result, nine Eimeria species (E.acervulina, E. brunetti, E. hagani, E. maxima, E. mitis, E. mivati, E. necatrix, E. praecox and E. tenella) have been reported to diagnosed in chickens (22, 23). In this study, we aimed to identify the Eimeria species causing coccidiosis in broilers using a molecular technique based on the Quantitative Real-Time PCR and determine the prevalence of coccidial infections in commercial broiler flocks in chickens.

Materials and Methods

1- Stool Sample collection From August 2013 to July 2014 a total of 200 samples of fresh fecal droppings and intestines were collected from suspected infected chickens with coccidiosis attending to the veterinary hospital and veterinary clinics were spread in Kerbala and Babylon provinces , Iraq for the examination and treatment . The oocysts were isolated from intestines and stool of infected chickens and collected in Eppendrof tubes and stored in freezing until used in DNA extraction . (12)

2- Direct examination method (microscopic examination): The first step in the present study is identified the positive samples from whole samples that collected from suspected chickens with coccidiosis and examined microscopically to determined the oocysts either by direct methods in cases of acute diarrhea in which the samples examined either freshly or used of a natural formalin buffer solution which concentrated 10% or by staining the samples with Iodin or Giemsa stain to made the internal components more clear.

3-Stool DNA Extraction Genomic DNA was extracted from stool samples of chicken by using AccuPrep® Stool DNA Extraction Kit (Bioneer, Korea)

4-DNA profile For detection of DNA that extract from stool samples through the use of a Nanodrop spectrophotometer (THERMO. USA) for detection and measurement the concentration of nuclear acids (DNA and RNA) ,Where is detected DNA concentration (ng / μl) and measuring the purity of the DNA by reading the absorbance at a wavelength of between (280-260 nm) ( 21).

5-qRT-PCR protocols Real-Time PCR was preformed for detection of poultry Eimeria species by using the primers and TaqMan probe specific for ITS1 region In the DNA that code for Ribosomal RNA, technique was carried out according to method described by(13).

Real-Time PCR master mix preparation Real-Time PCR master mix was prepared by one step Reverse Transcription and Real-Time PCR detection kit (AccuPower RocketScript RT-qPCR PreMix, Bioneer. Korea), and done according to company instructions as following Table (1):

Primers: The primers were designed in this study by using the complete sequence of ITS1 region in the rDNA using NCBI Gene-Bank and Primer3 plus online and provided by (Bioneer company, Korea) as showed in Table (2):

The qRT-PCR master mix reaction components that mentioned in (table 2 ) were added into standard qPCR tube containing (8 wells strips tubes which containing Rocket Script reverse transcriptase and TaqMan probe premix). Then all strips tubes vortexing for mixed the components and centrifuge for 3000 rpm for 3 minutes in Exispin centrifuge, after that transferred into Exicycler RealTime PCR thermocycler.

6-Real-Time PCR Thermocycler conditions Real-Time PCR thermocycler conditions was set up according to primer annealing temperature and RT-qPCR TaqMan kit instructions as following Table (3):

Thermal cycles were applied to inspect the Real-Time PCR and relying on instructions AccuPower® 2X GreenStarTM qPCR Master Mix as well as by calculating the degree Tm prefixes using the device MiniOpticon RealTime PCR system BioRad / USA as in Figure (1) below:

7-Real-Time PCR Data analysis qRT-PCR data analysis was performed by calculation the threshold cycle number (CT value) that presented the positive amplification of gene in Real-time cycle number.

Statistical analysis The results of present study analyzed statistically by Program The Statistical Analysis (SAS) by using of the Lest Significant Difference (LSD) test and Duncan test depending on the level of probability P <0.05 to find the significant differences (24).

Results

Results of molecular examination by qRT-PCR The results of molecular examination by using of qRTPCR revealed that from 200 collected samples appears 160(80%) were positive ,depending on this technique three species of Eimeria were diagnosed including E. tenella which form the highest percentage (66.8%) in comparison to other species ,While the E. necatrix form (30% )of the total positive samples and the last species that diagnosed was E. maxima which form ( 3.13%) as in the table(4).

The used of qRT-PCR techniques in the specific detection of Eimeria species showed a fluorescence of SYBER Green dye which appeared very clear through formed of amplification plot to the positive samples during the cycle 30 as shows in the fig.(2,3 and 4).

Discussion

In chickens is clearly central to a better understanding of epidemiology and dynamics of the disease in intensive and extensive chicken establishments. This is particularly important for planning an effective prevention and control program of coccidiosis. Traditionally, diagnosis has been achieved by detecting Eimeria oocysts excreted in the feces of chickens by measuring oocyst and sporocyst size or notes of the site and pathological lesions in the intestine of chickens. Although the microscopic examinations can absolutely show the positive fecal samples, but the traditional methods very limited in the specific diagnosis or identification of Eimeria species also These methodes are unreliable in mix infection when there are a multiple species of Eimeria infect a single host and there is Similarity in the size , shape of oocysts and the sites of infection (25). During recent years, there have been significant advances in the development of molecular-diagnostic tools like the Polymerase chain Reactions(PCR) and the Real-time PCR, to comparison between both of them : the qRT- PCR has several advantages: there is no need to post-amplification analysis, which reduces risks of contamination; the ability to better differentiate between different species of Eimeria infections; and numerical results which are much easier to distinguish than the visualization of a stained gel in the conventional PCR(14) . Several studies that used of PCR technique targeting different regions of the Eimeria genome,such as the 5S rRNA (the small subunit rRNA (12, 13), the sporozoite antigen gene EASZ240/160 (14) and ITS-1 (8, 15,17) and ITS-2 (18-20) genomic regions. Since the ITS regions are less conserved than the rRNA genes, big variations in this region of DNA sequence between the Eimerian species , makes the design of primers straight forward and reduces the risk of cross reactions among different species (6). The REAL-TIME assay have been shown to be directly comparable in terms of sensitivity and robustness, capable of detecting 10 parasite genomes but not one, unaffected by the presence of DNA derived from the host or the other test species (26). Every sporulated oocyst contains eight eimerian genomes, suggesting that the DNA equivalent of a single oocyst will be consistently detectable given normal experimental replication (between one and 10 genomes detected per reaction).Mature intracellular stages represent in the order of 10 to 100 eimerian genomes (depending on the species and stage (27). Suggesting that even a fraction of one may be counted (25). Apart from an accurate identification of Eimeria species, molecular methods can also be helpful in epidemiological study of the parasite, an aspect that has been less investigated to date at yet, there has not been any documentary report related to the specific identification and epidemiological pattern of the pathogenic Eimeria species in the chickens, in Karbala and Babylon provinces. Therefore, the results of the present study are the first study depend on molecular methods to detection the Eimeria species the causal agent of poultry coccidiosis in Kerbala and Babylon provinces. To compared the results of the present study with the resultes of researchers in various Arab and foreign countries ,in which the study which was conducted in Arab Republic of Egypt has been diagnosed prevalence of five species of Eimeria mainly include of E. tenella then E. necatrix, E. acervulina, E. maxima and finally type E .mitis using a technique REAL TIME- PCR (28). Researcher Muhammad, L. et al held a study on the diagnosis of infection in poultry coccidiosis t in Jordan using technique Quantitative REAL TIME- PCR and identified five species of Eimeria where the species E. tenella comes in front and reached a rate of about 80.5%, while the E. maxima (66%), E. necatrix (45%), E. acevulina (12%) and finally the species E. mitis (3.5%) (29). Also this technique were used in the Kingdom of Saudi Arabia to diagnosed the infection with coccidiosis in chickens, in which the stool samples collected from six fields scattered across the Kingdom and identified of four species of Eimeria in which the species E. necatrix appeared high precentage then the species E. tenella, E. maxima and E. acervulina (10). In comparison of the present study with the results of studies in foreign countries, including Australia, where the researcher use the REAL TIME PCR for the detection and diagnosis of seven species of Eimeria which infect chickens and targeted one genetic area which is called the ITS2 located in the ribosomal DNA (rDNA) using Taqman probs found that among the seven species the most prevalent species are the species E. tenella compared with the other six species (30). As in Japan used the technique SYBR Green-based RealTime PCR for the diagnosis of five species of Eimeria using Primers sets were designed based on the sequences area ITS1 in Ribosomal DNA specific to each species of the five and the researcher concluded that Melting Curves for each species has one Peak and the values of Melting temperature are specific for each species .The researcher concluded that the species E. brunett is the most prevalent species after E. maxima then E. necatrix E. tenella and finally the species E. acervulina ,so the REAL- TIME PCR technique is not only easy but also fast and efficient in qualitative diagnosis of poultry coccidiosis (31). It also conducted some studies on the diagnosis of poultry coccidiosis in a manner REAL- TIME PCR in some developing countries such as Ethiopia ,held a study on the prevalence of species of Eimeria in Ethiopian villages, by using this technique were reached to diagnosis of seven species of Eimeria where the percentage of species E. praecox was 95% and then species E. maxima (60%) and then species E. mitis (28%) and then come successively species E. tenella, E. necatrix and E. brunette (32). Carried out a diagnostic study is the first of its kind in Bangladesh for poultry coccidiosis following the molecular methods such as REAL TIME PCR technique .Where diagnosed seven species of Eimeria the most prevalent are E. tenella then E. necatrix , E. acervulina , E. maxima, E. brunetti, E. mitis and finally the species E. praecox (16). Efforts towards educating the chicken farmers to control coccidiosis in through good management practices, and the proper use of anticoccidial drugs should be considered. However, to control this economically important parasitic disease of poultry, further studies need to be undertaken to devise sustainable and cost-effective prevention and control methods

Conclusion

The following conclusions were conducted from the present study: 1-Although the routine tests such as clinical examination, macroscopic and microscopic diagnosis important, but it is unable to qualitative diagnosis of Eimeria that cause infection. 2- Using of the molecular methods such as REAL-TIME PCR which characterized by highly accuracy, but these methods expensive economically compared to routine methods. 3-The use of specific primers for diagnosis of ITS1 region are important in molecular detection of Eimeria species that isolated from intestines and feces of suspected chickens infection with Coccidiosis. 4- The E. tenella consider the most prevalent species (66.8%), Then E. necatrix (30%) and finally E. maxima which represented (3.13%). 5- The coccidiosis among chicken farms of Kerbala and Babylon is highly prevalent. E.tenella was the most prevalent species

. Acknowledgement

Authors acknowledgement the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to author’s publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed. Also authors would like to thanks department of biology, college of education for pure sciences, university of Kerbala for their support and encouragement. Funding and conflicting: There was no source of funding for this work and there was no any confliction of interest.

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A Study by Karimova II et al. entitled "Changes in the Activity of Intestinal Carbohydrases in Alloxan-Induced Diabetic Rats and Their Correction with Prenalon" is awarded Best Article of Vol 13 issue 22
A Study by Ashish B Roge et al. entitled "Development, Validation of RP-HPLC Method and GC MS Analysis of Desloratadine HCL and It’s Degradation Products" is awarded Best Article of Vol 13 issue 21
A Study by Isha Gaurav et al. entitled "Association of ABO Blood Group with Oral Cancer and Precancer – A Case-control Study" is awarded Best Article for Vol 13 issue 20
A Study by Amr Y. Zakaria et al. entitled "Single Nucleotide Polymorphisms of ATP-Binding Cassette Gene(ABCC3 rs4793665) affect High Dose Methotrexate-Induced Nephrotoxicity in Children with Osteosarcoma" is awarded Best Article for Vol 13 issue 19
A Study by Kholis Ernawati et al. entitled "The Utilization of Mobile-Based Information Technology in the Management of Dengue Fever in the Community Year 2019-2020: Systematic Review" is awarded Best Article for Vol 13 issue 18
A Study by Bhat Asifa et al. entitled "Efficacy of Modified Carbapenem Inactivation Method for Carbapenemase Detection and Comparative Evaluation with Polymerase Chain Reaction for the Identification of Carbapenemase Producing Klebsiella pneumonia Isolates" is awarded Best Article for Vol 13 issue 17
A Study by Gupta R. et al. entitled "A Clinical Study of Paediatric Tracheostomy: Our Experience in a Tertiary Care Hospital in North India" is awarded Best Article for Vol 13 issue 16
A Study by Chandran Anand et al. entitled "A Prospective Study on Assessment of Quality of Life of Patients Receiving Sorafenib for Hepatocellular Carcinoma" is awarded Best article for Vol 13 issue 15
A Study by Rosa PS et al. entitled "Emotional State Due to the Covid – 19 Pandemic in People Residing in a Vulnerable Area in North Lima" is awarded Best Article for Vol 13 issue 14
A Study by Suvarna Sunder J et al. entitled "Endodontic Revascularization of Necrotic Permanent Anterior Tooth with Platelet Rich Fibrin, Platelet Rich Plasma, and Blood Clot - A Comparative Study" is awarded Best Article for Vol 13 issue 13
A Study by Mona Isam Eldin Osman et al. entitled "Psychological Impact and Risk Factors of Sexual Abuse on Sudanese Children in Khartoum State" is awarded Best Article for Vol 13 issue 12
A Study by Khaw Ming Sheng & Sathiapriya Ramiah entitled "Web Based Suicide Prevention Application for Patients Suffering from Depression" is awarded Best Article for Vol 13 issue 11
A Study by Purushottam S. G. et al. entitled "Development of Fenofibrate Solid Dispersions for the Plausible Aqueous Solubility Augmentation of this BCS Class-II Drug" is awarded Best article for Vol 13 issue 10
A Study by Kumar S. et al. entitled "A Study on Clinical Spectrum, Laboratory Profile, Complications and Outcome of Pediatric Scrub Typhus Patients Admitted to an Intensive Care Unit from a Tertiary Care Hospital from Eastern India" is awarded Best Article for Vol 13 issue 09
A Study by Mardhiah Kamaruddin et al. entitled "The Pattern of Creatinine Clearance in Gestational and Chronic Hypertension Women from the Third Trimester to 12 Weeks Postpartum" is awarded Best Article for Vol 13 issue 08
A Study by Sarmila G. B. et al. entitled "Study to Compare the Efficacy of Orally Administered Melatonin and Clonidine for Attenuation of Hemodynamic Response During Laryngoscopy and Endotracheal Intubation in Gastrointestinal Surgeries" is awarded Best Article for Vol 13 issue 07
A Study by M. Muthu Uma Maheswari et al. entitled "A Study on C-reactive Protein and Liver Function Tests in Laboratory RT-PCR Positive Covid-19 Patients in a Tertiary Care Centre – A Retrospective Study" is awarded Best Article of Vol 13 issue 06 Special issue Modern approaches for diagnosis of COVID-19 and current status of awareness
A Study by Gainneos PD et al. entitled "A Comparative Evaluation of the Levels of Salivary IgA in HIV Affected Children and the Children of the General Population within the Age Group of 9 – 12 Years – A Cross-Sectional Study" is awarded Best Article of Vol 13 issue 05 Special issue on Recent Advances in Dentistry for better Oral Health
A Study by Alkhansa Mahmoud et al. entitled "mRNA Expression of Somatostatin Receptors (1-5) in MCF7 and MDA-MB231 Breast Cancer Cells" is awarded Best Article of Vol 13 issue 06
A Study by Chen YY and Ghazali SRB entitled "Lifetime Trauma, posttraumatic stress disorder Symptoms and Early Adolescence Risk Factors for Poor Physical Health Outcome Among Malaysian Adolescents" is awarded Best Article of Vol 13 issue 04 Special issue on Current Updates in Plant Biology to Medicine to Healthcare Awareness in Malaysia
A Study by Kumari PM et al. entitled "Study to Evaluate the Adverse Drug Reactions in a Tertiary Care Teaching Hospital in Tamilnadu - A Cross-Sectional Study" is awarded Best Article for Vol 13 issue 05
A Study by Anu et al. entitled "Effectiveness of Cytological Scoring Systems for Evaluation of Breast Lesion Cytology with its Histopathological Correlation" is awarded Best Article of Vol 13 issue 04
A Study by Sharipov R. Kh. et al. entitled "Interaction of Correction of Lipid Peroxidation Disorders with Oxibral" is awarded Best Article of Vol 13 issue 03
A Study by Tarek Elwakil et al. entitled "Led Light Photobiomodulation Effect on Wound Healing Combined with Phenytoin in Mice Model" is awarded Best Article of Vol 13 issue 02
A Study by Mohita Ray et al. entitled "Accuracy of Intra-Operative Frozen Section Consultation of Gastrointestinal Biopsy Samples in Correlation with the Final Histopathological Diagnosis" is awarded Best Article for Vol 13 issue 01
A Study by Badritdinova MN et al. entitled "Peculiarities of a Pain in Patients with Ischemic Heart Disease in the Presence of Individual Combines of the Metabolic Syndrome" is awarded Best Article for Vol 12 issue 24
A Study by Sindhu Priya E S et al. entitled "Neuroprotective activity of Pyrazolone Derivatives Against Paraquat-induced Oxidative Stress and Locomotor Impairment in Drosophila melanogaster" is awarded Best Article for Vol 12 issue 23
A Study by Habiba Suhail et al. entitled "Effect of Majoon Murmakki in Dysmenorrhoea (Usre Tams): A Standard Controlled Clinical Study" is awarded Best Article for Vol 12 issue 22
A Study by Ghaffar UB et al. entitled "Correlation between Height and Foot Length in Saudi Population in Majmaah, Saudi Arabia" is awarded Best Article for Vol 12 issue 21
A Study by Siti Sarah Binti Maidin entitled "Sleep Well: Mobile Application to Address Sleeping Problems" is awarded Best Article for Vol 12 issue 20
A Study by Avijit Singh"Comparison of Post Operative Clinical Outcomes Between “Made in India” TTK Chitra Mechanical Heart Valve Versus St Jude Mechanical Heart Valve in Valve Replacement Surgery" is awarded Best Article for Vol 12 issue 19
A Study by Sonali Banerjee and Mary Mathews N. entitled "Exploring Quality of Life and Perceived Experiences Among Couples Undergoing Fertility Treatment in Western India: A Mixed Methodology" is awarded Best Article for Vol 12 issue 18
A Study by Jabbar Desai et al. entitled "Prevalence of Obstructive Airway Disease in Patients with Ischemic Heart Disease and Hypertension" is awarded Best Article for Vol 12 issue 17
A Study by Juna Byun et al. entitled "Study on Difference in Coronavirus-19 Related Anxiety between Face-to-face and Non-face-to-face Classes among University Students in South Korea" is awarded Best Article for Vol 12 issue 16
A Study by Sudha Ramachandra & Vinay Chavan entitled "Enhanced-Hybrid-Age Layered Population Structure (E-Hybrid-ALPS): A Genetic Algorithm with Adaptive Crossover for Molecular Docking Studies of Drug Discovery Process" is awarded Best article for Vol 12 issue 15
A Study by Varsha M. Shindhe et al. entitled "A Study on Effect of Smokeless Tobacco on Pulmonary Function Tests in Class IV Workers of USM-KLE (Universiti Sains Malaysia-Karnataka Lingayat Education Society) International Medical Programme, Belagavi" is awarded Best article of Vol 12 issue 14, July 2020
A study by Amruta Choudhary et al. entitled "Family Planning Knowledge, Attitude and Practice Among Women of Reproductive Age from Rural Area of Central India" is awarded Best Article for special issue "Modern Therapeutics Applications"
A study by Raunak Das entitled "Study of Cardiovascular Dysfunctions in Interstitial Lung Diseas epatients by Correlating the Levels of Serum NT PRO BNP and Microalbuminuria (Biomarkers of Cardiovascular Dysfunction) with Echocardiographic, Bronchoscopic and HighResolution Computed Tomography Findings of These ILD Patients" is awarded Best Article of Vol 12 issue 13 
A Study by Kannamani Ramasamy et al. entitled "COVID-19 Situation at Chennai City – Forecasting for the Better Pandemic Management" is awarded best article for  Vol 12 issue 12
A Study by Muhammet Lutfi SELCUK and Fatma entitled "Distinction of Gray and White Matter for Some Histological Staining Methods in New Zealand Rabbit's Brain" is awarded best article for  Vol 12 issue 11
A Study by Anamul Haq et al. entitled "Etiology of Abnormal Uterine Bleeding in Adolescents – Emphasis Upon Polycystic Ovarian Syndrome" is awarded best article for  Vol 12 issue 10
A Study by entitled "Estimation of Reference Interval of Serum Progesterone During Three Trimesters of Normal Pregnancy in a Tertiary Care Hospital of Kolkata" is awarded best article for  Vol 12 issue 09
A Study by Ilona Gracie De Souza & Pavan Kumar G. entitled "Effect of Releasing Myofascial Chain in Patients with Patellofemoral Pain Syndrome - A Randomized Clinical Trial" is awarded best article for  Vol 12 issue 08
A Study by Virendra Atam et. al. entitled "Clinical Profile and Short - Term Mortality Predictors in Acute Stroke with Emphasis on Stress Hyperglycemia and THRIVE Score : An Observational Study" is awarded best article for  Vol 12 issue 07
A Study by K. Krupashree et. al. entitled "Protective Effects of Picrorhizakurroa Against Fumonisin B1 Induced Hepatotoxicity in Mice" is awarded best article for issue Vol 10 issue 20
A study by Mithun K.P. et al "Larvicidal Activity of Crude Solanum Nigrum Leaf and Berries Extract Against Dengue Vector-Aedesaegypti" is awarded Best Article for Vol 10 issue 14 of IJCRR
A study by Asha Menon "Women in Child Care and Early Education: Truly Nontraditional Work" is awarded Best Article for Vol 10 issue 13
A study by Deep J. M. "Prevalence of Molar-Incisor Hypomineralization in 7-13 Years Old Children of Biratnagar, Nepal: A Cross Sectional Study" is awarded Best Article for Vol 10 issue 11 of IJCRR
A review by Chitra et al to analyse relation between Obesity and Type 2 diabetes is awarded 'Best Article' for Vol 10 issue 10 by IJCRR. 
A study by Karanpreet et al "Pregnancy Induced Hypertension: A Study on Its Multisystem Involvement" is given Best Paper Award for Vol 10 issue 09

List of Awardees

A Study by Ese Anibor et al. "Evaluation of Temporomandibular Joint Disorders Among Delta State University Students in Abraka, Nigeria" from Vol 13 issue 16 received Emerging Researcher Award


A Study by Alkhansa Mahmoud et al. entitled "mRNA Expression of Somatostatin Receptors (1-5) in MCF7 and MDA-MB231 Breast Cancer Cells" from Vol 13 issue 06 received Emerging Researcher Award


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International Journal of Current Research and Review (IJCRR) provides platform for researchers to publish and discuss their original research and review work. IJCRR can not be held responsible for views, opinions and written statements of researchers published in this journal

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