Terrestrial specimens were collected from Ashshafa, a south-western highland area in Saudi Arabia. Three species, i.e., Porcellio laevis, Porcellionides pruinosus (Porcellionidae), and Armadillidium vulgare (Armadillidiidae), were identified in this study based on their morphological characteristics. Partial mitochondrial cytochrome C oxidase subunit I (COI) gene sequences were used for DNA barcoding and biodiversity assessments. A phylogenetic tree of 22 haplotypes from 35 specimens of the three isopod species was drawn from the most similar sequences obtained from BLAST with the associated accession numbers. The tree included two clades. The first clade included samples of P. laevis and P. pruinosus, whereas the second clade included samples of A. vulgare. Each identified species formed a distinct subclade within the main clade, along with similar sequences obtained from the NCBI database. The heat map of genetic distance among haplotypes shows the haplotype diversity (Hd) ranged from 0.590 to 0.933 (mean = 0.767) and total nucleotide diversity (πT) ranged from 0.001 to 0.089 (mean=0.049), with a similar trend observed for nucleotide diversity per site (θw) ranged from 0.001 to 0.80 (mean = 0.049). In contrast, synonymous nucleotide diversity (πs), mean=0.009, was low compared to nonsynonymous nucleotide diversity (πs), mean=0.060, across all species. In conclusion, the morphological identification of terrestrial isopods was confirmed using COI gene sequencing of mitochondrial DNA. These results will be helpful in developing a deeper isopod identification method.

Alzheimer's disease (AD) is a brain disorder and the main reason for dementia. In this regard, there is a need to understand the alterations that occur during aging to develop treatment strategies to mitigate or prevent neurodegenerative consequences. Onion and garlic root extracts contain natural polyphenols with high antioxidant capacity; therefore, the present study aimed to investigate the protective effect of these extracts free from mycotoxin contamination on a rat model of AD. Antifungal and antibacterial assays were performed for onion and garlic extracts. Several groups of AD-induced rats were administered 1, 2, and 3 mg/kg onion or garlic extract through intragastric intubation for 30 days. After treatment, histopathological analysis, expression of apoptosis-related genes, and analyses of DNA damage and reactive oxygen species (ROS) generation were conducted in the brain tissues. The results indicate that treatment of AD-induced rats with several doses of onion and garlic root extracts decreased histopathological lesions, the expression levels of apoptotic genes, and the rate of DNA damage and inhibited intracellular ROS generation in the brain tissues. The results suggest that the protective role of onion root extract could be attributed to its content of flavonoids and flavonoid compounds through the improvement of antioxidant capacity and regulation of gene expression patterns. The higher activity levels of free radical scavenging of azino-bis (3-ethylbenzothiazoline-6-sulphonic acid (ABTS) and antioxidant ferric reducing antioxidant power (FRAP) levels found in garlic root extract are most probably responsible for its protective effect against neurodegenerative damage.

Objectives: The purpose of the current study was to verify the reliability of COI bar- codes in the assessment of genetic diversity of two ecotypes from different ecozones. Methods: The DNA sequences of cytochrome oxidase I (COI) barcodes of 50 hens belonging to two ecotypes of Ismalia Egypt (ISM) and Taif Saudi Arabia (TA) were isolated and analyzed. Results: This study results showed that no noticeable great differences among all barcode’s sequences of both ecotypes. The aver- age length of both ecotypes was 589 bp. ISM ecotypes have a relatively wider length range. The overall mean of GC% content was 48±0.01. Both ecotypes have the same number of sites 548 bp. ISM ecotype has 523 monomorphic sites whereas TA ecotype has slightly fewer monomorphic sites 517. The ISM ecotype has 7 singleton sites and 18 Parsimony informative sites. TA ecotype has little more polymorphic, that is 12 singleton sites and 19 Par- simony informative sites. The number of mutations (η) was larger in ISM (46) compared to 38 mutations for TA ecotype. Both ecotypes had the same number of Haplotypes (25), and haplotypes diversity (1) as well as the variance of haplotype diversity. Conclusion: These results indicated a comparable level of genetic diversity of both ecotypes, which in turn may refer to a similarity of evolutionary forces that affect both ecotypes. Based on the present results, COI gene can be used in barcoding. The COI provides an objective the foundation for identification of ecotypes and therefore could be used for a rapid establishment of a variety of identifications.

The current study is the first comprehensive investigation to address the native chicken ecotypes of the Taif region to unravel the genetic diversity using a dense panel of 40 microsatellites (SSR). Blood samples were collected from 25 hens randomly sampled from a village farm at Taif governorate. A total of 147 alleles were detected, with an average of 3.7 alleles per locus. The overall mean of polymorphic information content (PIC) was 0.43. The average observed heterozygosity (Hobs) of 0.28 was lower than the expected heterozygosity (Hexp) of 0.48. Out of 40l ocionly11 loci showed insignificant deviation from Hardy Weinberg expectation. The ecotypes showed low genetic diversity (HS = 0.65) and a high level of inbreeding (FIS= 0.75). The high FIS is indicative of the endangerment potentiality of this ecotype. Nine SSR showed an inbreeding coefficient of one. The significant estimate of the inbreeding coefficient of the present study calls for an immediate breeding plan to preserve such endangered ecotypes. Results of the present study will provide an initial guide to design further investigations for the development of sustainable genetic improvement and conservation programs for the Taif ecotype genetic resources.

Several reasons may underlie the dramatic increase in type2 diabetes mellitus. One of these reasons is the genetic basis and variations. Vitamin D receptor polymorphisms are associated with different diseases such as rheumatoid arthritis and diabetes. The aim of this study is to investigate the possible association of two identified mutations ApaI (rs7975232) and TaqI (rs731236). Eighty-nine healthy individuals and Fifty-six Type 2 Diabetic (T2D) patients were investigated using RFLP technique for genotyping and haplotyping as well. The distribution of Apal genotypes was not statistically significant among the control (P=0.65) as well as for diabetic patients (P=0.58). For Taql allele frequencies of T allele was 0.61 where of G allele was 0.39. The frequency distribution of Taql genotypes was not statistically significant among the control (P=0.26) as well as diabetic patients (P=0.17). Relative risk of the allele T of Apa1 gene is 1.28 and the odds ratio of the same allele is 1.53, while both estimates were < 1.0 of the allele G. Similarly, with the Taq1 gene the relative risk and the odds ratio values for the allele T are 1.09 and 1.27 respectively and both estimates of the allele C were 0.86 for the relative risk and 0.79 for the odds ratio. The pairwise linkage disequilibrium between the two SNPs Taq1/apa1 was statistically significant in control group (D = 0.218, D' = 0.925 and P value < 0.001) and similar data in diabetic groups (D = 0.2, D' = 0.875 and P value < 0.001). These data suggest that the T allele of both genes Apa1 and Taq1 is associated with the increased risk of type 2 diabetes. We think that we need a larger number of volunteers to reach a more accurate conclusion.

Aim: To assess genetic diversity of two chicken ecotypes from Egypt and Saudi Arabia and compare these ecotypes to six local Egyptian and two exotic pure chicken breeds using 14 microsatellite markers. Methodology: Dataset consisted of two subsets. First subset represented two ecotypes from Saudi Arabia and Egypt. Second dataset consisted birds six Egyptian native chicken strains: Fayoumi (FAY), Dandarawy (DAN), Baladi (BAL), Sinai (Sini), El-Salam (Els) and Golden Montazah (GG), and two commercial strains: White Leghorn (WL) and Rhode Island Red (RIR). fourteen microsatellites markers were utilized to assess DNA polymorphism. Data analyses of the results were carried out using R statistical environment. Results: The obtained results indicated that number of alleles per locus averaged 11.4 ± 5.0. Polymorphic information content was informative (> 50%) for the local breeds, but not for two ecotypes. The observed and expected heterozygosity averaged 0.46 and 0.75, both ecotypes had the lowest estimates. All breeds showed significant deviation from Hardy–Weinberg equilibrium expectation. The average population differentiation index (FST) was 0.143, overall heterozygosity deficiency (FIT) was 0.156, and global inbreeding of individuals within breeds (FIS) was 0.319. Interpretation: This study revealed that both Egyptian and Saudi ecotypes have an endangered status. Lower genetic distances and lower FST values were reported for the Egyptian indigenous breeds. Phylogenetic and principle components showed that both ecotypes were genetically closer to each other when compared with other breeds. It also showed that the Dandarawy native Egyptian chicken breed was genetically the closest breed to both the Egyptian and Saudi ecotypes. Key words: Chicken, Ecotypes, Genetic diversity, Microsatellites, Phylogenetics

Genetic diversity of two chicken ecotypes from Ismailia-Egypt (ISM) and Taif-Saudi Arabia (TA) was evaluated using 39 microsatellites. DNA was extracted from blood of 25 chickens/ecotype. The number of alleles was 157 and 138, the number of alleles/locus averaged 4.2±2.2 and 3.6±1.6, and the highest number of private alleles was 9 and 5 for ISM and TA, respectively. Percentage of shared alleles between the two ecotypes was 45%. This panel of markers is reasonably informative as the mean polymorphic information content for ISM and TA was 0.47±0.21, and 0.41±0.2. Similar average of observed heterozygosity was attained for both ecotypes. Conversely, averages of expected heterozygosity differed between two ecotypes, 0.52±0.23 vs. 0.45±0.21 for ISM and TA. 8 and 12 loci have significantly deviated from HWE of ISM and TA. Estimate of genetic distance was 0.2 and FST index was 0.29. Results showed only 6% of genetic diversity is shared between these two ecotypes.

The high nutritional value of fish makes it highly perishable because it provides favorable medium for the growth of micro-organisms after catching. In addition, they can also act as carriers of several microbial and other health hazards. Many mould growth on foods stored at low temperature is common and recurring problem. Certain molds are known to be capable of producing mycotoxins at low temperature as low as (-2 to 10 degrees) C. That encouraging us to perform this study which aimed to survey, isolation and perform different further genetic identification of fungal spoilage species of some marine fish from Saudi Arabia markets. 100 samples from 5 types of fishes; Salmon "Salmoniformes", Red sea Bream "Pagrus pagrus", Rabbitfish "Siganus rivulatus", Spanish mackerel "Scomberomorous commerson", Red mullet "Mullus surmuletus", collected from Jeddah fish markets, then prepared aseptically and plated on Potato Dextrose Agar (PDA) medium incubated for 3-7 days/28 +/- 2 degrees C and examined daily Macroscopically, microscopically and genetically. The results recorded 4 species of fungi; Aspergillus eucalypticola as one of (Black Aspergilli), Aspergillus oryzae as (white Aspergilli), Penicillium digitatum as (green fungi) and Byssochlamys spectabilis. This is the first report of Penicillium digitatum and Byssochlamys spectabilis in fish. Furthermore, the research identified each fungi gene cluster. More attention in fish rearing facilities, caution should be taken by consumers in preparation and applying perfect cooking in consuming fish and more education and efforts should be developed by fish farmers to avoid fishponds contamination. We recommended to further research should be done on fungal contaminations.

Isolating microbes from diverse natural ecological units has led to achieve metabolite structural diversity. Hot springs have been less explored ecological sects for discovery of novel microbial bioactive compounds as compared to other terrestrial samples. The capability of thermophilic microorganisms to flourish at high temperatures makes their enzyme systems ideal for various biotechnological applications. In this study, a total of 11 bacterial strains were isolated from Al-Lith hot spring which named Oyun Al-Haar, located about 17 km northeast of Gomika, at about 250 km south of Jeddah, Saudi Arabia. All these isolates were characterized and screened for some enzymes production. They were able to abundantly grow on starch nitrate agar medium and grew optimally at 450 with pH 7.0. According to bioinformatics analysis, the eleven bacterial isolates were encompassing 9 actinomycetes and 2 eubacteria namely, Streptomyces tendae (3 isolates) and only one isolate for each of Streptomyces mutabilis, Streptomyces chitinivorans, Streptomyces barkulensis, Leclercia adecarboxylata, Streptomyces fradiae, Streptomyces azureus, Streptomyces macrosporus and Enterobacter cloacae. Based on the enzymatic activities, all isolates were positive for keratinase, gelatinase, chitinase, and lipase. Nine out of the eleven isolates showed protease production in good levels, and out of these11 isolates, 6 isolates exhibited remarkable amylase activity. Bacteria, especially actinomycetes isolated from hot springs area have gained commercial significance as source of thermostable enzymes. There are few reports of enzymes production from the microflora of hot springs, which opens a window for exploring this resource as potential cache of novel strains with bioactive compounds.

36 Bacillus native strains were isolated and identified using 16s gene sequencing analysis. Universal primer of 16s rRNA gene was used and PCR amplifications were done with all DNA isolation from all isolated strains. Mixture of non-identical 16S rRNA amplicons products produced and bioinformatics data analysis of all yielded variable consensus sequence were done using different software programs. The data analysis of sequences displayed some sequence variability occurs among the multiple 16S rRNA genes. Bioinformatical analysis revealed that the nucleotide sequences obtained from the isolates were different homologous to the known DNA sequence using NCBI Blast software. Results revealed that the obtained Bacillus isolated strains were belonging to17 different species. No noticeable base frequencies variation were observed in base frequencies for all the 39 isolates except B.simplex strains in in cytosine base. Results of phylogenic analysis revealed that all isolated strains are clustered analysis in 2 large clusters. The Polymorphism and Genetic Diversity among the isolates were also done using Clustalx and Maft build analysis, the monomorphic and polymorphic sits of all strains also analyzed.Further study for complete genome sequence analysis will be needed. The bioinformatics tools were succeeded in analyze all our native bacillus strains.

In the present study, 30 multidrug-resistant bacterial samples were isolated from different hospitals in the Taif governorate in Saudi Arabia. Given its discriminating power as a universal phylogenetic marker, the 16S rDNA gene was sequenced in a comprehensive diversity study to determine the evolutionary and phylogenetic relationships among the bacterial isolates. The 16S rDNA genes of all isolates were successfully amplified using PCR, and comprehensive identification results were based on GenBank databases. Analysis revealed nucleotide identities ranging from 76% to 100% based on the consensus sequences of 21 species, namely, Bacillus cereus, Bacillus subtilis, Bacillus tequilensis, Caldimonas manganoxidans, Citrobacter freundii, Enterococcus faecium, Escherichia fergusonii, Klebsiella pneumoniae, Lactobacillus plantarum, Lactococcus garvieae, Leuconostoc mesenteroides, Myristica yunnanensis, Pantoea eucrina, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus capitis, Staphylococcus caprae, Staphylococcus epidermidis, Staphylococcus hominis, Staphylococcus petrasii, and Staphylococcus saccharolyticus. We observed high variability in terms of DNA length and GC content between and within species. Phylogenetic analysis clustered the isolates into three groups. The number of sites ranged from 827 (S. aureus) to 1,219 (L. mesenteroides). Estimation of nucleotide diversity ((partial derivative) over bar) showed that all analyzed sequences were diverse site-wise and also exhibited high nucleotide diversity, with ((partial derivative) over bar) values ranging from 0.17 to 0.94. All isolates showed significantly conserved regions (P>0.05). In conclusion, the observed variations in the sequences of the target bacterial strains can be attributed to resistance to antibiotics and gene transfer among bacterial strains in the hospital environment. Further sequence analyses of antibiotic resistance genes are warranted.

In order to assess and characterize genetic diversity of Saudi Arabia native chickens, a total of 25 individuals were genotyped using panel of12 microsatellites (MS) markers, and the highly polymorphic MS markers were selected based on the highest value of the number of alleles. Twenty-three alleles were amplified using microsatellites primers with score-able fragment sizes ranging from approximately 110 to 343bp. fourteen alleles were found polymorphic, revealing 60.8% polymorphism rate. The genetic distance(GD) was estimated according to microsatellites data, estimated GD value ranged between 35.6 and 88. The highest GD value was 88.2 between chicken sample no. 5 and chicken sample no. 8, while the lowest GD was 35.6 between chicken sample no. 10 and chicken sample no. 26. Neighbor-joint method clustering grouped samples into two groups, where the two chicken samples no. 10 and 23 were clustered in different group. Thus this penal of microsatellites markers provide good insight into the genetic diversity available across Saudi indigenous chickens of either close or distant genetic backgrounds. It also could be concluded that Saudi indigenous chickens possess the needed amount of genetic variation required for further genetic improvement of these breeds.

Cadmium (Cd) is the most prevalent toxic metal present in livestock feed; therefore, the present study aimed to examine the ameliorative effects of grape seed extract (GSE) on cadmium chloride (CdCl2)‑induced testicular dysfunction of Wistar rats. Male adult Wistar rats (40 rats; n=10/group) were divided into four equal groups. Group one was used as a control, and was given ad libitum access to food and water. Groups 2‑4 were treated with CdCl2 [5 mg/kg body weight (BW)], GSE (400 mg/kg BW, orally), and GSE plus CdCl2, respectively. Blood and testicular tissues were collected and assayed for biochemical and histopathological changes, respectively. Testicular genes were expressed using semi‑quantitative RT‑PCR analysis. The results of the present study demonstrated that there was a decrease in serum testosterone levels following CdCl2 toxicity, which were normalized after GSE co-administration. Furthermore, CdCl2 significantly increased the serum levels of malondialdehyde, and decreased levels of antioxidants. At the histopathological level, the testes of the CdCl2 group exhibited congestion, edema in the interstitial blood vessels, irregular arrangement of the epithelial lining of the seminiferous tubules, and degeneration and sloughing of the spermatogenic cells, which accumulated in the center of the seminiferous tubules. Such pathological alterations were ameliorated following treatment with GSE in the CdCl2 plus GSE group. The immunohistochemical expression of B‑cell lymphoma 2‑associated X protein was high in the CdCl2 group, and low in the control and GSE groups. Co‑treatment with GSE and CdCl2 exhibited ameliorative effects on the immunoreactivity of B‑cell lymphoma 2‑associated X protein. CdCl2 toxicity induced a significant downregulation in the mRNA expression levels of cytochrome P450 cholesterol side‑chain cleavage enzyme, cytochrome P450 17A1, 3β‑hydroxysteroid dehydrogenase (3β‑HSD), 17β‑HSD, androgen receptor, steroidogenic acute regulatory protein, and follicle‑stimulating hormone receptor. GSE administration exhibited a stimulatory effect on steroidogenesis‑associated enzymes, and co‑treatment with GSE and CdCl2 normalized and upregulated the mRNA expression levels of these examined genes. This study concluded that GSE has beneficial protective effects against the deleterious effects of CdCl2 on the testis.

The Taif domestic chicken (Gallus gallus, 2n = 78) is believed to have descended from the wild Indian and Southeast Asian red jungle fowl. The molecular characterization of local chickens in different districts of Taif region in Saudi Arabia was done using RAPD markers of the variable region. The twelve random primers were used to generate fingerprint patterns for these chickens. Twenty five DNA of blood samples from individual chicken were tested to confirm the fingerprinting and genetic distance among these chickens. Specific results for fingerprinting were obtained by the twelve primers retained for RAPD analysis produced different fragment patterns with varied number of bands. The primers yielded a total of 187 distinct bands 34.7% were considered as polymorphic bands and 65.3% were considered as monomorphic bands. The OPA-06 primer has showed the highest polymorphism 83.3% and total of 18 bands ranged from 250 bp-1700 bp. While, the OPA-03 primer has showed the lowest polymorphism 7.8% and total of 14 bands ranged from 200 bp-1650bp.The Dendrogram based on RAPD results grouped the twenty five individual local chicken samples into two different clusters with about 80% genetic similarity. Moreover, the genetic distance among native chicken was relatively low. The smallest genetic distance (0.09) was estimated between sample No. 23 and sample No. 19.This work aims to estimate the genetic resources in the local Saudi chickens reared using RAPD markers in Taif region as preliminary work to established the basis for genetic conservation program for local chicken in Saudi Arabia.

This study aimed to evaluate the effect of altitude on some male fertility related traits in Saudi sheep and goats. Testicular tissues were collected from a slaughterhouse in Taif governorate (1800 m above sea level) and Makkah governorate (sea level). Sperm characteristics (1 - individual motility, 2 - percent live sperm 3 - percent abnormal sperm) were examined. Semi-quantitative RT - PCR assay was used to evaluate the expression of IGF-II, StAR, LDLr and CYP11A genes. No significant effect of altitude on tested sperm parameters was revealed. Expression of IGF-II gene in both sheep and goats was significantly (P<0.05) higher at sea level compared to high altitude. A similar effect of altitude on StAR gene expression was only observed in goats, while in sheep the level of effect did not reach the significance threshold. Moreover, LDLr gene expression was significantly (P<0.05) higher for both sheep and goats at high altitude than at sea level. The CYP11A gene expression was significantly (P<0.05) higher in high altitude sheep than in those raised at sea level, while the opposite trend was observed for goats. In conclusion, high altitude had an effect on the expression of some studied male fertility related genes, but sperm parameters were not significantly affected.

The present work aims to molecularly characterize some popular Saudi fish species. Seven popular Saudi fish species namely, Morgan (Nemipteru sjaponicus), Mousa (Solea solea), Hamor (Greasy grouper), Shour (Lethrinus lentjan), Dennis (Caranxsex fasciatus), Harid (Scarus arabicus) and Black surgeon (Acanthurus gahhm) were characterized using six RAPD and three ISSR as well as Rep-PCR markers. These markers produced 187 distinct bands 89.4% were considered as polymorphic bands and 10.6% were considered as monomorphic bands. The RAPD OPA-06 primer revealed 100% polymorphism for 19 produced bands where band width ranged from 190 to 2100 bp. Likewise Rep- PCR ERIC1R primer exhibited 100% polymorphism for 18 bands produced bands where band width ranged from 50 to 1400bp. Data of the three types of genetic markers were combined for phylogenetic analysis. The resulted dendergram produced two large linages with around 59% genetic similarity. One linage only included Morgan (Nemipteru sjaponicus), and the other linage comprised the other six species. This result suggested that this six species were descended from Morgan fish. The second linage comprised two clades; the first clade contained four species (Mousa, Hamor, Shour, and Dennis), where the next clade included only Harid and Black surgeon. Generally genetic distance among native fishes was relatively low. The smallest genetic distance (0.512) was estimated between Shour fish and Black surgeon fish. To the best of our knowledge this work is breaking new ground in two directions, first, molecular characterization of Saudi fish, second employment Rep- PCR genetic marker for molecular characterization of fish species. This work could be considered as preliminary work towards an establishment of Saudi genetic conservation program.Int J Appl Sci Biotechnol, Vol 3(2): 359-366 DOI: http://dx.doi.org/10.3126/ijasbt.v3i2.12747

The genus Chaetomium is a rich source of novel and bioactive secondary metabolites of great importance. To date, a variety of more than 200 secondary metabolites belonging to diverse structural types have been discovered. Fungal enzymes are used in food, beverages, confectionaries, textiles, and leather industries to simplify the processing of raw materials. They are often more stable than enzymes derived from other sources. Ten isolates of Chaetomium globosum recovered and designated as TUCg1 to TUCg10 were identified by morphological and molecular biology means and submitted to the GenBank. These isolates were screened for extracellular enzymes such as amylase, cellulase, laccase, lipase, pectinases, protease and chitinase on solid media. All Chaetomium globosum isolates screened for potential enzymes showed amylolytic, cellulolytic, and proteolytic activities; six isolates were chitinolytic and laccase producers; and five and three isolates showed pectinolytic and lipolytic activities, respectively. The produced array of enzymes differed among isolates. Molecular techniques such as internal transcribed spacer (ITS) region sequencing and specific genes random primers polymerase chain reaction (SGRP-PCR) have shown high DNA polymorphism of Chaetomium globosum. In conclusion, SGRP-PCR is a rapid and valuable tool for assessment and characterization of genetic diversity of Chaetomium globosum, which suggests the use of this technique for identification of different fungal isolates.

6 Abstract: The prevalence of diabetes is high among the Saudi population and 90% of diabetics suffer from Type II diabetes mellitus (T2DM). Genetic polymorphisms are integral to the development of genetic markers to identify individuals at risk of developing T2DM. These genetic markers could be used for early detection and molecular diagnosis of diabetes. The objective of the present study is to detect the prevalence of A6224G polymorphism within the Insulin receptor substrate-1 (IRS-1) gene and tRNA)Leu(UUR) 3243 polymorphism in mitochondrial DNA among T2DM. Saudi patients. One hundred and six patients were recruited. Genotyping was carried out via PCR-RFLP technique. No patients with tRNA)Leu(UUR) 3243 polymorphisms were found. Three genotypes of A6224G polymorphism were detected. The prevalence of the detected genotypes was (40.6 for AA, 58.5 for AG and 0.94 for GG and ). The frequency of A allele was 0.7 and for G allele was 0.3.