Studies on the antiperoxide and antihaemolytic action of polyamines using human erythrocytes

Blood RBC's were used to study the role of polyamines, putrescine, spermidine and spermine individually as antioxidants against the peroxidative action of H₂O₂ which leads to haemolysis of erythrocytes. Results indicated that putrescine did not act as antioxidant at all concentrations used in our studies. However, spermidine and spermine suppressed peroxidative changes in RBC's due to the action of H₂O₂ and this suppression was greater with higher concentration of these individual polyamines used, separately. Further, all the three polyamines decreased haemolysis of erythrocytes at all concentrations studied. The action of putrescine to suppress haemolysis of RBC's without showing antioxidant property hypothetically suggests that these properties of respective polyamines may have independent mechanism of action.     

Molecular characterization and genetic diversity of different genotypes of Oryza sativa and Oryza glaberrima

BackgroundAvailability of related rice species is critical for rice breeding and improvement. Two distinct species of domesticated rice exist in the genus Oryza: Oryza sativa (Asian rice) and Oryza glaberrima (African rice). New rice for Africa (NERICA) is derived from interspecific crosses between these two species. Molecular profiling of these germplasms is important for both genetics and breeding studies. We used 30 polymorphic SSR markers to assess the genetic diversity and molecular fingerprints of 53 rice genotypes of O. sativa, O. glaberrima, and NERICA.ResultsIn total, 180 alleles were detected. Average polymorphism information content and Shannon's information index were 0.638 and 1.390, respectively. Population structure and neighbor-joining phylogenetic tree revealed that 53 genotypes grouped into three distinct subpopulations conforming to the original three groups, except three varieties (IR66417, WAB450-4, MZCD74), and that NERICA showed a smaller genetic distance from O. sativa genotypes (0.774) than from O. glaberrima genotypes (0.889). A molecular fingerprint map of the 53 accessions was constructed with a novel encoding method based on the SSR polymorphic alleles. Ten specific SSR markers displayed different allelic profiles between the O. glaberrima and O. sativa genotypes.ConclusionsGenetic diversity studies revealed that 50 rice types were clustered into different subpopulations whereas three genotypes were admixtures. Molecular fingerprinting and 10 specific markers were obtained to identify the 53 rice genotypes. These results can facilitate the potential utilization of sibling species in rice breeding and molecular classification of O. sativa and O. glaberrima germplasms.     

Genetic diversity among wild pomegranate (Punica granatum) in Azad Jammu and Kashmir region of Pakistan

BackgroundPomegranate (Punica granatum L.), one of the most important tropical fruits in Azad Jammu and Kashmir regions of Pakistan, is highly valued for its nutrition and medicinal purposes. Although pomegranate is native to this region, the genetic diversity among wild pomegranate accessions is currently unknown. Such information would be vital for germplasm conservation and breeding efforts. In the current study, genetic diversity among forty-eight wild pomegranate accessions collected from different agro-ecological zones of Azad Jammu and Kashmir was assessed using 41 simple sequence repeat (SSR) markers.ResultsThe markers revealed 303 alleles averaging 7.39 alleles per marker. Polymorphic information content ranged from 0.12 (PGCT093B) to 0.88 (Pom006), with a mean of 0.54. The average genetic distance (GD) across all genotypes was 0.52, and was lowest between Chattar Class and Thorar genotypes (GD = 0.27), but highest between Khun Bandway and Akhor Ban (GD = 0.74). A neighbor-joining dendrogram separated the genotypes into three major clusters, with further sub-clustering within each cluster.ConclusionsOverall, the results presented here show significant genetic diversity among wild pomegranate accessions in Azad Jammu and Kashmir region of Pakistan. These accessions present a valuable genetic resource to breeding and cultivar improvement programs within the region.How to cite: Aziz S, Firdous S, Rahman H, et al. Genetic diversity among wild pomegranate (Punica granatum) in Azad Jammu and Kashmir region of Pakistan. Electron J Biotechnol 2020;46. https://doi.org/10.1016/j.ejbt.2020.06.002.     

Segregation distortion in homozygous lines obtained via anther culture and maize doubled haploid methods in comparison to single seed descent in wheat (Triticum aestivum L.)

BackgroundThe quality of wheat grain depends on several characteristics, among which the composition of high molecular weight glutenin subunits, encoded by Glu-1 loci, are the most important. Application of biotechnological tools to accelerate the attainment of homozygous lines may influence the proportion of segregated genotypes. The objective was to determine, whether the selection pressure generated by the methods based on in vitro cultures, may cause a loss of genotypes with desirable Glu-1 alleles.ResultsHomozygous lines were derived from six winter wheat crosses by pollination with maize (DH-MP), anther culture (DH-AC) and single seed descent (SSD) technique. Androgenetically-derived plants that originated from the same callus were examined before chromosome doubling using allele-specific and microsatellite markers. It was found that segregation distortion in SSD and DH-MP populations occurred only in one case, whereas in anther-derived lines they were observed in five out of six analyzed combinations.ConclusionsSegregation distortion in DH-AC populations was caused by the development of more than one plant of the same genotype from one callus. This distortion was minimized if only one plant per callus was included in the population. Selection of haploid wheat plants before chromosome doubling based on allele-specific markers allows us to choose genotypes that possess desirable Glu-1 alleles and to reduce the number of plants in the next steps of DH production. The SSD technique appeared to be the most advantageous in terms of Mendelian segregation, thus the occurrence of residual heterozygosity can be minimized by continuous selfing beyond the F₆ generation.     

Analysis of genetic diversity and differentiation of sheep populations in Jordan

BackgroundGenetic diversity of sheep in Jordan was investigated using microsatellite markers (MS). Six ovine and bovine MS located on chromosomes 2 and 6 of sheep genome were genotyped on 294 individual from ten geographical regions.ResultsThe number of alleles per locus (A), the expected heterozygosity (H_e) and observed heterozygosity (H_o) were measured. Overall A, H_e and H_o were 12.67, 0.820 and 0.684, respectively. On the other hand, genetic distances undoubtedly revealed the expected degree of differentiation among the studied populations. The finding showed closeness of three populations from south (Maan, Showbak and Tafeilah) to each other. Populations from the middle regions of Jordan (Karak, Madaba, Amman, AzZarqa and Mafraq) were found to be in one cluster. Only two populations of the middle region were an exception: AlSalt and Dead Sea. Finally, sheep populations from Irbid were located in separated cluster. It was clear that the studied predefined populations were subdivided from four populations and would be most probably accounted as ancestral populations. These results indicate that number of population is less than the predefined population as ten based on geographical sampling areas.ConclusionsThe possible inference might be that geographical location, genetic migration, similar selection forces, and common ancestor account for population admixture and subdivision of Awassi sheep breed in Jordan. Finally, the present study sheds new light on the molecular and population genetics of Awassi sheep from different regions of Jordan and to utilize the possible findings for future management of genetic conservation under conditions of climate changes and crossbreeding policy.     

Production of wheat-doubled haploids resistant to eyespot supported by marker-assisted selection

BackgroundWheat is one of the most important crops cultivated all over the world. New high-yielding cultivars that are more resistant to fungal diseases have been permanently developed. The present study aimed at the possibility of accelerating the process of breeding new cultivars, resistant to eyespot, by using doubled haploids (DH) system supported by marker-assisted selection.ResultsTwo highly resistant breeding lines (KBP 0916 and KBH 4942/05) carrying Pch1 gene were crossed with the elite wheat genotypes. Hybrid plants of early generations were analyzed using endopeptidase EpD1 and two SSR markers linked to the Pch1 locus. Selected homozygous and heterozygous genotypes for the Pch1-linked EpD1b allele were used to produce haploid plants. Molecular analyses were performed on haploids to identify plants possessing Pch1 gene. Chromosome doubling was performed only on haploid plants with Pch1 gene. Finally, 65 DH lines carrying eyespot resistance gene Pch1 and 30 lines without this gene were chosen for the eyespot resistance phenotyping in a field experiment.ConclusionsResults of the experiment confirmed higher resistance to eyespot of the genotypes with Pch1 in comparison to those without this gene. This indicates the efficiency of selection at the haploid level.How to cite: Wiśniewska H, Majka M, Kwiatek M, et al. Production of wheat doubled haploids resistant to eyespot supported by marker-assisted selection. Electron J Biotechnol 2019;37. https://doi.org/10.1016/j.ejbt.2018.10.003     

Characterization of Lactobacillus fermentum UCO-979C,a probiotic strain with a potent anti-Helicobacter pylori activity

BackgroundHelicobacter pylori is considered as the main risk factor in the development of gastric cancer. In the present study, we performed a detailed characterization of the probiotic properties and the anti-H. pylori activity of a previously isolated lactobacillus strain — Lactobacillus fermentum UCO-979C — obtained from human gut.ResultsThe strain tolerated pH 3.0; grew in the presence of 2% bile salts; produced lactic acid and hydrogen peroxide; aggregated in saline solution; showed high hydrophobicity; showed high adherence to glass; Caco-2 and gastric adenocarcinoma human cells (AGS) cells; showed an efficient colonization in Mongolian Gerbils; and potently inhibited the growth and urease activity of H. pylori strains. L. fermentum UCO-979C significantly inhibited H. pylori-induced IL-8 production in AGS cells and reduced the viability of H. pylori. With regard to innocuousness, the strain UCO-979C was susceptible to several antibiotics and did not produce histamine or beta-haemolysis in blood agar containing red blood cells from various origins.ConclusionThe results demonstrated that L. fermentum UCO-979C is a very good candidate as a probiotic for the protection of humans against H. pylori infections.     

Development and application of KASP marker for high throughput detection of AhFAD2 mutation in peanut

BackgroundCultivated peanut (Arachis hypogaea L.) is a major oilseed crop worldwide. Fatty acid composition of peanut oil may affect the flavor and shelf life of the resulting food products. Oleic acid and linoleic acid are the major fatty acids of peanut oil. The conversion from oleic acid to linoleic acid is controlled by the Δ12 fatty acid desaturase (FAD) encoded by AhFAD2A and AhFAD2B, two homoeologous genes from A and B subgenomes, respectively. One nucleotide substitution (G:C → A:T) of AhFAD2A and an “A” insertion of AhFAD2B resulted in high-oleic acid phenotype. Detection of AhFAD2 mutation had been achieved by cleaved amplified polymorphic sequence (CAPS), real-time polymerase chain reaction (qRT-PCR) and allele-specific PCR (AS-PCR). However, a low cost, high throughput and high specific method is still required to detect AhFAD2 genotype of large number of seeds. Kompetitive allele specific PCR (KASP) can detect both alleles in a single reaction. The aim of this work is to develop KASP for detection AhFAD2 genotype of large number of breeding materials.ResultsHere, we developed a KASP method to detect the genotypes of progenies between high oleic acid peanut and common peanut. Validation was carried out by CAPS analysis. The results from KASP assay and CAPS analysis were consistent. The genotype of 18 out of 179 BC₄F₂ seeds was aabb.ConclusionsDue to high accuracy, time saving, high throughput feature and low cost, KASP is more suitable for determining AhFAD2 genotype than other methods.     

Identification and genetic diversity analysis of Memecylon species using ISSR,RAPD and Gene-based DNA barcoding tools

BackgroundMemecylon species are commonly used in Indian ethnomedical practices. The accurate identification is vital to enhance the drug's efficacy and biosafety. In the present study, PCR based techniques like RAPD, ISSR and DNA barcoding regions, such as 5s, psbA-trnH, rpoC1, ndh and atpF-atpH, were used to authenticate and analyze the diversity of five Memecylon species collected from Western Ghats of India.ResultsPhylogenetic analysis clearly distinguished Memecylon malabaricum from Memecylon wightii and Memecylon umbellatum from Memecylon edule and clades formed are in accordance with morphological keys. In the RAPD and ISSR analyses, 27 accessions representing five Memecylon species were distinctly separated into three different clades. M. malabaricum and M. wightii grouped together and M. umbellatum, M. edule and Memecylon talbotianum grouped in the same clade with high Jaccard dissimilarity coefficient and bootstrap support between each node, indicating that these grouped species are phylogenetically similar.ConclusionData from the present study reveals that chloroplast psbA-trnH region could be used as a potential candidate region for identifying Memecylon species, and ISSR marker system could be used for estimating genetic diversity since it has high percent polymorphism compared to RAPD marker.     

The molecular diversity analysis of Auricularia auricula-judae in China by nuclear ribosomal DNA intergenic spacer

BackgroundFor the crossbreeding of Auricularia auricula-judae, selecting the appropriated parents in hybridization is very important. However, the classification and diversity analysis of A. auricula-judae has been equivocal, due to the similarity of the fruiting body morphology and its susceptibility to environmental influences. For this purpose, the molecular diversity of 32 A. auricula-judae commercial cultivars in China was analyzed by using the nuclear ribosomal DNA intergenic spacer.ResultsThe complete nuclear rDNA gene complex of A. auricula-judae isolate is 11,210 bp long, and contains the 18S, 5.8S, and 28S rRNA gene as well as the ITS and IGS regions. Based on the sequence data, four more effective primer combinations for the IGS region of A. auricula-judae were designed. Nucleotide sequence variation in the IGS among 32 A. auricula-judae commercial cultivars in China sorted into three strongly supported clades, which is correlated with geographical regions. Most strains originated from the same area were with a narrow genetic basis and could possibly be domesticated from the local wild-type strains.ConclusionThe grouping information obtained in the present work provides significant information for further genetic improvement in A. auricula-judae, and suggested that the IGS region can be used as an excellent tool for identification of genetic variation.     

Genome-wide development of polymorphic microsatellite markers and their application in peanut breeding program

BackgroundCultivated peanut (Arachis hypogaea. L) represents one of the most important oil crops in the world. Although much effort has been expended to characterize microsatellites or Simple Sequence Repeats (SSRs) in peanut, the quantity and quality of the markers in breeding applications remain limited. Here, genome-wide SSR characterization and marker development were performed using the recently assembled genome of the cultivar Tifrunner.ResultsIn total, 512,900 microsatellites were identified from 2556.9-Mb genomic sequences. Based on the flanking sequences of the identified microsatellites, 7757 primer pairs (markers) were designed, and further evaluated in the assembled genomic sequences of the tetraploid Arachis cultivars, Tifrunner and Shitouqi, and the diploid ancestral species, A. duranensis and A. ipaensis. In silico PCR analysis showed that the SSR markers had high amplification efficiency and polymorphism in four Arachis genotypes. Notably, nearly 60% of these markers were single-locus SSRs in tetraploid Arachis species, indicating they are more specific in distinguishing the alleles of the A and B sub-genomes of peanut. In addition, two markers closely related with purple testa color and 27 markers near to FAD2 genes were identified, which could be used for breeding varieties with purple testa and high-oleic acid content, respectively. Moreover, the potential application of these SSR markers in tracking introgressions from Arachis wild relatives was discussed.ConclusionsThis study reported the development of genomic SSRs from assembled genomic sequences of the tetraploid Arachis Tifrunner, which will be useful for diversity analysis, genetic mapping and functional genomics studies in peanut.How to cite: Ma J, Zhao Y, Chen H, et al. Genome-wide development of polymorphic microsatellite markers and their application in peanut breeding program. Electron J Biotechnol 2020;44. https://doi.org/10.1016/j.ejbt.2020.01.004.     

Analysis of the genetic relationships and diversity among 11 populations of Xanthoceras sorbifolia using phenotypic and microsatellite marker data

BackgroundAssessments of genetic diversity are essential for germplasm characterization and exploitation. Molecular markers are valuable tools for exploring genetic variation and identifying germplasm. They play key roles in a Xanthoceras sorbifolia breeding program.ResultsWe analyzed the genetic diversity of populations of this species using 23 simple sequence repeat (SSR) loci and data on kernel oil content. The 11 populations included in the study were distributed across a large geographic range in China. The kernel oil content differed significantly among populations. The SSR marker analysis detected high genetic diversity among the populations. All SSRs were polymorphic, and we identified 80 alleles across the populations. The number of alleles at each locus ranged from two to six, averaging 3.48 per primer pair. The polymorphism information content values ranged from 0.35 to 0.70, averaging 0.51. Expected heterozygosity, observed heterozygosity, and Shannon's information index calculations detected large genetic variations among populations of different provenance. The high average number of alleles per locus and the allelic diversity observed in the set of genotypes analyzed indicated that the genetic base of this species was relatively wide. The statistically significant positive correlation between genetic and geographic distances suggested adaptations to local conditions.ConclusionsMicrosatellite markers can be used to efficiently distinguish X. sorbifolia populations and assess their genetic diversity. The information we have provided will contribute to the conservation and management of this important plant genetic resource.     

Pleiotropic effects of the sdw1 locus in barley populations representing different rounds of recombination

BackgroundIn the present study populations, representing different rounds of recombination were used for the analysis of phenotypic effects associated with the sdw1/denso locus. Other studies have mostly focused only on one type of population. Many different QTLs mapped at the same position as the sdw1/denso locus may indicate a pleiotropy of this gene or a tight linkage between genes conditioning quantitative traits. To date, results of studies have not unequivocally proven either of these two phenomena.ResultsBoth breeding and molecular mapping experiments were undertaken to examine 200 single seed descent (SSD) and 60 doubled haploid (DH) lines obtained from the Maresi (with a semi-dwarfing gene) and Pomo cross combination. They were evaluated for the type of juvenile growth habit and certain agronomic traits were measured after harvesting. The estimates of mean values, standard errors and significance of effects were analyzed. In terms of the analyzed characteristics, the greatest variability was obtained for genotypes with the prostrate growth habit. Microsatellite markers (SSR) were also used to identify co-segregation with the sdw1/denso locus and Bmag0013, Bmag0877, Bmag0306b markers were linked the closest. A partial linkage map of chromosome 3H with the sdw1/denso semi-dwarfing gene was constructed and QTLs were identified.ConclusionsOur experiments confirmed the impact of the semi-dwarfing gene on plant height, heading and flowering date both in SSD and DH populations, which may indicate pleiotropy. Moreover, a partial linkage between sdw1/denso locus and grain weight per spike and 1000-grain weight was found in the SSD population.     

Expression profiles and polymorphism analysis of CDIPT gene on Qinchuan cattle

BackgroundCDIPT (CDP-diacylglycerol–inositol 3-phosphatidyltransferase, EC 2.7.8.11) was found on the cytoplasmic side of the Golgi apparatus and the endoplasmic reticulum. It was an integral membrane protein performing the last step in the de novo biosynthesis of phosphatidylinositol (PtdIns). In recent years, PtdIns has been considered to play an essential role in energy metabolism, fatty acid metabolic pathway and intracellular signal transduction in eukaryotic cells.ResultsIn this study, the results of real-time polymerase chain reaction (PCR) showed that the expression of CDIPT gene was remarkably different in diverse tissues. We also detected the polymorphism of bovine CDIPT gene and analyzed its association with body measurement and meat quality traits of Qinchuan cattle. Blood samples were obtained from 638 Qinchuan cattle aged from 18 to 24 months. DNA sequencing and PCR-restriction fragment length polymorphism (RFLP) were used to find CDIPT gene single nucleotide polymorphism (SNP). Three SNPs g.244T>C (NCBI: rs42069760), g.1496G>A and g.1514G>A were found in this study. g.244T>C located at 5′untranslated region (5′UTR) of exon 1 showed three genotypes: TT, TC and CC. g.1496G>A and g.1514G>A detected the first time were located in intron 3 and showed the same genotypes: GG, GA and AA.ConclusionsAnalysis results showed that these three SNPs were significantly associated with body measurement traits (BMTs) and meat quality traits (MQTs). We suggested that CDIPT gene may have potential effects on BMTs and MQTs and can be used for marker-assisted selection.     

Indole acetic acid and ACC deaminase from endophytic bacteria improves the growth of Solanum lycopersicum

BackgroundEndophytic bacteria are ubiquitous in all plant species contributing in host plant's nutrient uptake and helping the host to improve its growth. Moringa peregrina which is a medicinal plant, growing in arid region of Arabia, was assessed for the presence of endophytic bacterial strains.ResultsPCR amplification and sequencing of 16S rRNA of bacterial endophytes revealed the 5 endophytic bacteria, in which 2 strains were from Sphingomonas sp.; 2 strains from Bacillus sp. and 1 from Methylobacterium genus. Among the endophytic bacterial strains, a strain of Bacillus subtilis LK14 has shown significant prospects in phosphate solubilization (clearing zone of 56.71 mm after 5 d), ACC deaminase (448.3 ± 2.91 nM α-ketobutyrate mg^- 1 h^- 1) and acid phosphatase activity (8.4 ± 1.2 nM mg^- 1 min^- 1). The endophytic bacteria were also assessed for their potential to produce indole-3-acetic acid (IAA). Among isolated strains, the initial spectrophotometry analysis showed significantly higher IAA production by Bacillus subtilis LK14. The diurnal production of IAA was quantified using multiple reactions monitoring method in UPLC/MS–MS. The analysis showed that LK14 produced the highest (8.7 μM) IAA on 14th d of growth. Looking at LK14 potentials, it was applied to Solanum lycopersicum, where it significantly increased the shoot and root biomass and chlorophyll (a and b) contents as compared to control plants.ConclusionThe study concludes that using endophytic bacterial strains can be bio-prospective for plant growth promotion, which might be an ideal strategy for improving growth of crops in marginal lands.     

Narcissism and entrepreneurial intentions: The roles of entrepreneurial self-efficacy and environmental complexity

PurposeExploring factors that can influence employee's entrepreneurial intentions is critical to entrepreneurship and entrepreneurial practice. This study is going to examine the effect of narcissism on entrepreneurial intentions. Moreover, the mediating role of entrepreneurial self-efficacy in narcissism-entrepreneurial intentions link is also tested. Lastly, how environmental complexity moderates these relationships is explored.Design/methodology/approachData were collected from 362 employees working in technology incubators and science parks of Saudi Arabia. These technology incubators and science parks were established by the Saudi government as a part of entrepreneurship ecosystem evolution strategy to encourage and support entrepreneurial innovations.FindingsThe results show that narcissistic employees display greater levels of entrepreneurial intentions, and entrepreneurial self-efficacy mediates the effect of narcissism on entrepreneurial intentions; and environmental complexity moderates the narcissism-entrepreneurial intentions link.OriginalityThe effect of narcissism on entrepreneurial intentions has not been examined before. This study extends knowledge on entrepreneurial innovations by linking narcissism with entrepreneurial intentions through mediation of entrepreneurial self-efficacy and moderation of environmental complexity.     

Genetic variability of Sargassum liebmannii on the coast of Jalisco in the central Mexican Pacific revealed by molecular markers and morphological traits

BackgroundSargassum liebmannii is widely distributed throughout rocky, coastal upwelling areas in the tropical Mexican Pacific. This brown algae is of great environmental and industrial importance. However, no information is available that documents the genetic or phenotypic variability of the species, which is needed to determine how it may react to environmental variation related to climate change. In this study, S. liebmannii specimens were collected from the coast of Jalisco, Mexico, and molecular and morphological characterization was conducted. Intraspecific variability was estimated according to the study areas.ResultsThe inter-simple sequence repeat (ISSR) markers indicated a polymorphism percentage of 95%. The Shannon index and Nei index showed relatively low values among the populations (0.3569 and 0.081, respectively). On the other hand, the genetic differentiation coefficient indicated inter- and intrapopulation values of 36.69% and 63.31%, respectively. The Jaccard similarity coefficient was used to determine the degree of similarity among individuals by geographical area. The morphological characteristics and environmental variables that were used to correlate phenotypes and genotypes indicated that S. liebmannii showed low genetic flow because of the presence of geographical barriers due to substrate that was not optimal for algal development.ConclusionsThe ISSR markers were useful for detecting genetic differences among S. liebmannii individuals. The results indicate that a coupled genotypic-phenotypic study is beneficial for documenting the variation present in the little-studied algal species. These studies may be used in future research to clarify taxonomic controversies while generating additional genomic information.How to cite: Jung-Kim HW, Hernández-Herrera RM, Enciso-Padilla I, et al. Genetic variability of Sargassum liebmannii on the coast of Jalisco in the central Mexican Pacific revealed by molecular markers and morphological traits. Electron J Biotechnol 2021;54. https://doi.org/10.1016/j.ejbt.2021.08.003     

Google trends identifying seasons of religious gathering: applied to investigate the correlation between crowding and flu outbreak

In medical research, mass gathering is considered as one of the intensifying factors for respiratory disease outbreaks. An annual crowding event happens during Muslims’ religious practices of Hajj in Saudi Arabia. Another smaller ritual happens all year round, known as Umrah. While Hajj season is recognized, Umrah seasonality is challenging to identify. This paper uses data from Google Trends to identify Umrah seasons for performers from the Eastern Province in Saudi Arabia. It then investigates the correlation between mass gathering events as it affects the flu spread in the Eastern Province through the use of data. Patient data from a hospital was obtained to find flu seasonality and compared to crowding seasons. Google Trend data was confirmed by a limited dataset of official Umrah data and Flu data was confirmed by official FluNet data from neighboring country, Bahrain. An Exploratory Data Analysis (EDA) approach has been followed to identify the seasonality. Cross-correlation analysis found a strong positive correlation between crowding seasons and flu outbreaks in the Eastern Province with a forward shift of flu data for three months.