Respective Allele Frequencies Research Articles

Molecular Screening of β-Lactoglobulin Gene Variation for Allergy-free Milk Production in Indonesian Friesian Holstein

The most allergenic protein in cow’s milk for new-borns is Beta Lactoglobulin (β-LG), which is also one of the bovine whey proteins. As a result, choosing suitable animals to create free allergenic protein is essential. In this study, Indonesian Friesian Holstein’s genetic variants in the β-LG gene were examined). Forty-two blood samples were sampled as DNA sources. The β-LG gene allele and genotype variant were identified using PCR-RFLP, or Polymerase Chain Reaction - Restriction Fragment Length Polymorphism. Successful amplification of the β-LG gene with a 247 bp product size was followed by DNA sequencing and HaeIII enzyme digestion (GG|CC). There were discovered to be three genotypes and two types of alleles, A and B, with the frequencies being AA (0.26), AB (0.50), and BB (0.24). The population is in equilibrium because A and B have respective allele frequencies of 0.51 and 0.49. In conclusion, molecular-assisted selection can create allergen-free milk by utilizing the genotype variants of β-LG genes.

Sequence-based allelic variations and frequencies for 22 autosomal STR loci in the Lebanese population

This is the first study that characterizes the sequence-based allelic variations of 22 autosomal Short Tandem Repeat (aSTR) loci in a population dataset collected from Lebanon. Genomic DNA extracts from 195 unrelated Lebanese individuals were amplified with PowerSeq 46GY System Prototype. Targeted amplicons were subjected to DNA library preparation and sequenced on the Verogen MiSeq FGx Sequencing System. Raw FASTQ data files were processed by STRait Razor v3. Sequence strings were annotated according to the considerations of the DNA Commission of the International Society for Forensic Genetics (ISFG) and tabulated herein with their respective allelic frequencies and GeneBank accession and version numbers. The sequenced Lebanese dataset resulted in 429 distinct allelic sequences as compared to the 236 alleles identified by length only. The increase in the number of alleles was observed at 18 out of 22 aSTR loci and was attributed to the sequence variations residing in both the STR repeat motifs and flanking regions. The study uncovered 25 novel aSTR allelic sequences across 12 loci for which GenBank records did not previously exist in the STRSeq BioProject, PRJNA380127. For a concordance check, the length-based allelic calls derived from the full sequences were compared to those genotyped using capillary electrophoresis (CE) methods. Population genetic parameters relevant to the evaluation of forensic DNA evidence were assessed for the sequence-based data and compared to the parameters generated from the length-based information. Using the sequence-based data, Analysis of MOlecular VAriance (AMOVA), genetic distances, and population genetic structure were evaluated for 1231 individuals sampled from the Lebanese and four U.S. populations (African American, Asian, Caucasian, and Hispanic). The results were tabulated and visualized in a population tree, multidimensional scaling scatter plots, and bar plots. This newly established sequence-based database for the Lebanese population can be beneficial for extending NGS applicability to casework or paternity testing and assessing the strength of evidence for NGS-STR profiles. The described novel sequence variants at certain loci can further help in the effort to characterize the sequence diversity of STR markers from different populations around the world.

ASSOCIATION OF Nrf2 GENE POLYMORPHISM WITH POLYCYSTIC OVARY SYNDROME —A CASE-CONTROL STUDY

Background: Polycystic Ovary Syndrome (PCOS) is considered as one of the most common endocrine disorder in women of reproductive age. Exact cause of PCOS is still unknown. However, evidence for genetic basis has been reported. Reactive oxygen species and antioxidants have been documented as key factors involved in ovarian physiological metabolism. Nrf2 is a key transcription factor that regulates the expression of antioxidant proteins, therefore provides protection against oxidative stress. Objective of this study was to find the association of Nrf2 (rs6721961) gene polymorphism with pathogenesis of PCOS among Pakistani population. Methods: This case-control study was conducted in Pakistan Railway Hospital, Rawalpindi, from Oct 2020 to Sep 2021. The study included 200 PCOS patients diagnosed according to Rotterdam diagnostic criteria and 200 healthy controls. Blood samples of all participants were collected and DNA was extracted by Chelex Method. Polymerase Chain Reaction (PCR) was performed to find respective allelic frequencies of Nrf2 (rs6721961) genotype using specific primers. Results: The frequency of CC, CA and AA genotypes of rs6721961 polymorphism of Nrf2 gene were 72.5%, 27.5%, 0% in controls and 56.5%, 39.5%, 4% in cases. Significant association of CA genotype of Nrf2 gene polymorphism (rs6721961) and allele A were found with PCOS (OR: 0.54, 95% CI: 0.35–0.82, p=0.004), (OR: 0.61, 95% CI: 0.42–0.89, p=0.01) respectively. This is the first study to find out association of polymorphism in Nrf2 gene (6721961) with PCOS in any population. Conclusion: Nrf2 gene polymorphism (rs6721956) is significantly associated with PCOS among Pakistani population. Pak J Physiol 2022;18(3):15?8

Association of C(-1019)G with Risk of Development of Major Depression in Diabetics with Respect to Various Pakistani Ethnicities

Objective: To determine whether the ethnicity is associated with the risk of development of Major Depressive Disorder (MDD) in adult diabetic population with respect to the genetic association with single nucleotide polymorphism (SNP) C(-1019)G. Material and Methods: A total of 400 subjects were included in the study, out of which 200 were cases and 200 were age, gender and ethnicity matched healthy controls. Out of 200 cases , 100 cases had diabetes mellitus (DM) only and 100 cases had DM with MDD. Both males and females were included in the study having ages 25 years and above. Cases of both type I and type II DM were included in the study. Blood samples were collected and DNA was extracted by Chelax method. Real-time PCR was carried out to determine respective allelic frequencies of C(-1019)G genotype using TaqMan SNP genotyping assays and master mix. Results: According to our results no significant association was found between C(-1019)G genotype and risk of development of MDD in Pakistani diabetic population with respect to Ethnicity but diseased genotype was found more in Pathan population. Conclusion: There is no significant association of SNP C(-1019)G of 5-Hydroxytryptamine 1A receptor gene with MDD in Pakistani diabetics with respect to ethnicity. Keywords: Diabetes mellitus, major depressive disorder, 5-hydroxytryptamine 1A receptor gene, single nucleotide polymorphism, polymerase chain reaction.

Association of Nrf2 gene polymorphism (rs6721961) with semen parameters in primary male infertility cases in Pakistani population

Objective: To determine an association of single nucleotide polymorphism of the NRF2 gene (RS6721961) with semen parameters of primary male infertility patients in the Pakistani population.
 Study Design: Comparative cross-sectional study.
 Place and Duration of Study: Department of Biochemistry, Islamic International Medical College, Rawalpindi, in collaboration with two private infertility clinics (American Infertility Center, Rawalpindi and Mother and Child International Hospital and Research Institute, Mirpur, AJK) from Oct 2020 to Sep 2021.
 Methodology: A total of 288 participants were included in this study. There were 144 diagnosed cases of primary male infertility and 144 healthy fertile males, age and ethnicity matched controls. Blood samples were collected from participants after obtaining written informed consent. DNA was extracted by the Chelex TM Methodoogy. Multiplex PCR was done to determine the respective allelic frequencies of NRF2 (RS6721961) genotypes using specific primers.
 Results: There was no significant association between NRF2 genotypes and semen volume and semn colour in infertile males (p-value 0.32 and 0.84, respectively). Out of 144 cases, 111 (77%) had normal sperm count, 29 (20.1%) had oligospermia, while 4 (2.7%) patients had azoospermia. A significant association was observed between NRF2 genotypes and sperm concentration of infertile males (p-value <0.001). Out of 144 cases, 80 (55.5%) had normal motility, 57 (39.5%) had asthenospermia while 7 (8.4%) patients had necrospermia. A strong association was observed between NRF2 genotypes and sperm motility in infertile males (p-value <0.001).
 Conclusion: Single nucleotide polymorphism (RS6721961) of the NRF2 gene is significantly associated with sperm concentration and......

Association of Fok 1 (Rs10735810) Gene Polymorphism with Dental Caries in Pakistani Adolescents

Objective: To determine the association of single nucleotide polymorphism of vitamin D receptor gene Fok 1 (rs10735810) with dental caries in Pakistani adolescents.
 Study Design: Case-control study.
 Place and Duration of Study: Biochemistry Department, Islamic International Medical College, Rawalpindi in cooperation with Pathology Department, Railway General Hospital, Rawalpindi Pakistan, from Sep 2019 to Aug 2020.
 Methodology: A total of 304 participants were included in the study. There were 152 dental caries cases and 152 normal agematched controls. After taking written informed consent, blood samples were collected from each participant. DNA extraction was done by Chelex Method. PCR was carried out to determine the respective allelic frequencies of Fok 1 (rs10735810) genotype using specific primers.
 Results: Out of 304 study participants, there were 26 males (17.1%) and 126 females (82.9%) in case-group; and 37 males (24.3%) and 115 females (75.7%) in control-group. No significant association of Fok 1 (rs10735810) genotype with DMFT score was found in dental caries case-group. Regarding genotypic and allelic frequencies, there was no significant association of Fok 1 (rs10735810) genotype with dental caries case-group.
 Conclusion: Single nucleotide polymorphism of vitamin D receptor gene Fok 1 (rs10735810) is not associated with dental caries in Pakistani adolescents.

An Overview of the Use of Genotyping Techniques for Assessing Genetic Diversity in Local Farm Animal Breeds.

Simple SummaryThe number of local farm animal breeds is declining worldwide. However, these breeds have different degrees of genetic diversity. Measuring genetic diversity is important for the development of conservation strategies and, therefore, various genomic analysis techniques are available. The aim of the present work was to shed light on the use of these techniques in diversity studies of local breeds. In summary, a total of 133 worldwide studies that examined genetic diversity in local cattle, sheep, goat, chicken and pig breeds were reviewed. The results show that over time, almost all available genomic techniques were used and various diversity parameters were calculated. Therefore, the present results provide a comprehensive overview of the application of these techniques in the field of local breeds. This can provide helpful insights into the advancement of the conservation of breeds with high genetic diversity.Globally, many local farm animal breeds are threatened with extinction. However, these breeds contribute to the high amount of genetic diversity required to combat unforeseen future challenges of livestock production systems. To assess genetic diversity, various genotyping techniques have been developed. Based on the respective genomic information, different parameters, e.g., heterozygosity, allele frequencies and inbreeding coefficient, can be measured in order to reveal genetic diversity between and within breeds. The aim of the present work was to shed light on the use of genotyping techniques in the field of local farm animal breeds. Therefore, a total of 133 studies across the world that examined genetic diversity in local cattle, sheep, goat, chicken and pig breeds were reviewed. The results show that diversity of cattle was most often investigated with microsatellite use as the main technique. Furthermore, a large variety of diversity parameters that were calculated with different programs were identified. For 15% of the included studies, the used genotypes are publicly available, and, in 6%, phenotypes were recorded. In conclusion, the present results provide a comprehensive overview of the application of genotyping techniques in the field of local breeds. This can provide helpful insights to advance the conservation of breeds.

Association Between Collagenous and Lymphocytic Colitis and Risk of Severe Coronavirus Disease 2019

Association Between Collagenous and Lymphocytic Colitis and Risk of Severe Coronavirus Disease 2019

The frequency of combined IFITM3 haplotype involving the reference alleles of both rs12252 and rs34481144 is in line with COVID-19 standardized mortality ratio of ethnic groups in England.

Evidence was brought forward in England and the USA that Black, Asian, Latino and Minority Ethnic people exhibit higher mortality risk from COVID-19 than White people. While socioeconomic factors were suggested to contribute to this trend, they arguably do not explain the range of the differences observed, allowing for possible genetic implications. Almost concurrently, the analysis of a cohort in Chinese COVID-19 patients proposed an association between the severity of the disease and the presence of the minor allele of rs12252 of the Interferon-induced transmembrane protein 3 (IFITM3) gene. This SNP, together with rs34481144, are the two most studied polymorphisms of IFITM3 and have been associated in the past with increased severity in Influenza, Dengue, Ebola, and HIV viruses. IFITM3 is an immune effector protein that is pivotal for the restriction of viral replication, but also for the regulation of cytokine production. Following up on these two developments in the ongoing SARS-CoV-2 pandemic, the present study investigates a possible association between the differences in mortality of ethnic groups in England and the combined haplotypes of rs12252 and rs34481144. The respective allele frequencies were collected for 26 populations from the 1000 Genomes Project and subgroups were pooled wherever possible to create correspondences with ethnic groups in England. A significant correlation (r = 0.9687, p = 0.0003) and a striking agreement was observed between the reported Standardized Mortality Ratios and the frequency of the combined haplotype of both reference alleles, suggesting that the combination of the reference alleles of the specific SNPs may be implicated in more severe outcomes of COVID-19. This study calls for further focus on the role of IFITM3 variants in the mechanism of cellular invasion of SARS-CoV-2, their impact in COVID-19 severity and their possible implications in vaccination efficacy.

Glutathione S-Transferase Theta 1 / Mu 1 Null Genotype and Risk of Development of Actinic Keratosis in Pakistani Population.

To assess the frequency and association of Glutathione S-Transferase Theta 1 and Glutathione S-Transferase Mu 1 null genotypes in development of actinic keratosis (AK) in a group of Pakistani population. Case-control analytical study. Department of Biochemistry, Islamic International Medical College, Rawalpindi in collaboration with Department of Dermatology, Railway Hospital and Rural Health Center, District Health Office, Rawalpindi from September 2018 to September 2019. Methodology: A total of 86 participants were included in this study with 27 biopsy proven cases of AK and 59 matched controls. Blood samples were collected after obtaining written informed consent; and DNA was extracted by Chelex™ method. Multiplex PCR (M-PCR) was done to find respective allelic frequencies of GSTM1 and GSTT1 genes in both cases and controls. Results: Mean age of participants in cases and controls was 62.93 ±10.29 years and was 61.42 ±9.96 years, respectively. There were 18 males (66.7%) and 9 females (33.3%); and 43 males (72.9%) and 16 females (27.1%) in cases and controls, respectively. There was a significant association of GSTT1 null genotype with AK (OR: 2.72, 95% CI: 1.05-7.05, p = 0.037). There was a positive correlation between GSTT1 null genotype and AK (r = 0.225, p = 0.037). GSTT1 null genotype has a significant association for AK development in the studied Pakistani population. Key Words: Actinic keratosis, Glutathione S-Transferase Mu 1, Glutathione S-Transferase Theta 1, Polymerase chain reaction, Squamous cell carcinoma.

Genetic polymorphisms with erythrocyte traits in malaria endemic areas of Mali.

African populations are characterized by high degree of genetic diversity. This high genetic diversity could result from the natural selection pressure. Several studies have described an association between some genetic diversities and difference of susceptibility to infectious diseases like malaria. It seems therefore important to consider genetic diversity impact when interpreting results of clinical trials in malaria endemic areas. This study aimed to determine the genetic polymorphism with erythrocyte traits in different populations of malaria endemic area in Mali. The cross-sectional surveys were carried out in different ethnic groups living in malaria endemic areas in Mali. Six milliliters of whole blood were collected in EDTA vials from each participant after informed consent has been obtained. The ABO, RH, Kell, MNSs, Kidd and Duffy systems phenotypes were assessed by the technique of gel filtration. A total of 231 subjects were included from six villages. The blood groups phenotypes O (40.7%) and A (31.2%) were more frequent with respective allele frequencies of 0.65 and 0.21. In the RH system the haplotypes R0 (0.55), r (0.20) and R1 (0.13) were the most frequent. Seven percent (7%) of Duffy positive and 4% of Glycophorin B deficiency (S-s-) were observed among participants. All participants were Kell negative. ABO and RH systems were polymorphic in these ethnic groups in Mali. Their implication in susceptibility to malaria should be taken into account in clinical trials interpretation, and for prevention of blood transfusion risks during anemia frequently caused by malaria in children.

Determination of Polymorphisms in Pituitary Genes of the Native Afghani Naked Neck Chicken.

We investigated means to improve the production of the indigenous Naked Neck chicken in Afghanistan. Specifically, we analyzed single nucleotide polymorphisms (SNPs) in the prolactin (PRL) (24 bp indel), growth hormone (GH) (T185G), and pituitary specific transcript factor 1 (PIT-1) (intron 5) genes. Blood samples were collected from 52 birds and genomic DNA was extracted. Polymorphisms in the mentioned loci were analyzed by PCR, allele-specific PCR, and PCR-restriction fragment length polymorphism (RFLP) using TaqI and MspI endonucleases. Cloning followed by DNA sequencing was performed to ascertain the accuracy of the PCR-RFLP analysis for PIT-1.Two alleles were found for the PRL 24 bp indel, GH (T185G), and PIT-1/TaqI, with the following respective allelic frequencies: PRL-In 0.64 and PRL-Del 0.36, GH-T 0.91 and GH-G 0.09, and PIT-1-A 0.64 and PIT-1-B 0.36. Regarding the PIT-1/MspI polymorphism, three novel MspI recognition sites, as well as two reported MspI recognition sites, were detected in intron 5. Moreover, during sequence screening, two novel SNPs were found that generated restriction sites for MseI. Therefore, our results suggest that the PRL indel, GH T185G, and PIT-1/TaqI polymorphisms may be used as selection markers for Afghanistan Naked Neck chickens. Intron 5 of PIT-1 in the Afghani Naked Neck chicken was highly polymorphic compared to the reported Gallus gallus PIT-1 gene (GenBank accession no. NC_006088.4).

The dopamine-related polymorphisms BDNF, COMT, DRD2, DRD3, and DRD4 are not linked with changes in CSF dopamine levels and frequency of HIV infection.

We showed previously that higher levels in CSF dopamine in HIV patients are associated with the presence of the dopamine transporter (DAT) 10/10-repeat allele which was also detected more frequently in HIV-infected individuals compared to uninfected subjects. In the current study, we investigated further whether other genetic dopamine (DA)-related polymorphisms may be related with changes in CSF DA levels and frequency of HIV infection in HIV-infected subjects. Specifically, we studied genetic polymorphisms of brain-derived neurotrophic factor, catechol-O-methyltransferase, and dopamine receptors DRD2, DRD3, and DRD4 genetic polymorphisms in uninfected and HIV-infected people in two different ethnical groups, a German cohort (Caucasian, 72 individuals with HIV infection and 22 individuals without HIV infection) and a South African cohort (Xhosan, 54 individuals with HIV infection and 19 individuals without HIV infection). We correlated the polymorphisms with CSF DA levels, HIV dementia score, CD4+ T cell counts, and HIV viral load. None of the investigated DA-related polymorphisms was associated with altered CSF DA levels, CD4+ T cell count, viral load, and HIV dementia score. The respective allele frequencies were equally distributed between HIV-infected patients and controls. Our findings do not show any influence of the studied genetic polymorphisms on CSF DA levels and HIV infection. This is in contrast to what we found previously for the DAT 3'UTR VNTR and highlights the specific role of the DAT VNTR in HIV infection and disease.

A common functional variant on the pro-inflammatory Interleukin-6 gene may modify the association between long-term PM10 exposure and diabetes.

BackgroundAir pollutants have been linked to type 2 diabetes (T2D), hypothesized to act through inflammatory pathways and may induce interleukin-6 gene (IL6) in the airway epithelium. The cytokine interleukin-6 may impact on glucose homeostasis. Recent meta-analyses showed the common polymorphisms, IL6 -572G > C and IL6 -174G > C to be associated with T2D risk. These IL6 variants also influence circulatory interleukin-6 levels. We hypothesize that these common functional variants may modify the association between air pollutants and T2D.MethodsWe cross-sectionally studied 4410 first follow-up participants of the Swiss Cohort Study on Air Pollution and Lung and Heart Diseases (SAPALDIA), aged 29 to 73 years who had complete data on genotypes, diabetes status and covariates. We defined diabetes as self-reported physician-diagnosed, or use of diabetes medication or non-fasting glucose >11.1 mmol/L or HbA1c > 0.065. Air pollution exposure was 10-year mean particulate matter <10 μm in diameter (PM10) assigned to participants’ residences using a combination of dispersion modelling, annual trends at monitoring stations and residential history. We derived interaction terms between PM10 and genotypes, and applied mixed logistic models to explore genetic interactions by IL6 polymorphisms on the odds of diabetes.ResultsThere were 252 diabetes cases. Respective minor allele frequencies of IL6 -572G > C and IL6 -174G > C were 7 and 39 %. Mean exposure to PM10 was 22 μg/m3. Both variants were not associated with diabetes in our study. We observed a significant positive association between PM10 and diabetes among homozygous carriers of the pro-inflammatory major G-allele of IL6 -572G > C [Odds ratio: 1.53; 95 % confidence interval (1.22, 1.92); Pinteraction (additive) = 0.003 and Pinteraction (recessive) = 0.006]. Carriers of the major G-allele of IL6 -174G > C also had significantly increased odds of diabetes, but interactions were statistically non-significant.ConclusionsOur results on the interaction of PM10 with functionally well described polymorphisms in an important pro-inflammatory candidate gene are consistent with the hypothesis that air pollutants impact on T2D through inflammatory pathways. Our findings, if confirmed, are of high public health relevance considering the ubiquity of the major G allele, which puts a substantial proportion of the population at risk for the development of diabetes as a result of long-term exposure to air pollution.Electronic supplementary materialThe online version of this article (doi:10.1186/s12940-016-0120-5) contains supplementary material, which is available to authorized users.

The Prevalence of the Prothrombin (F2) 20210G>A Mutation in a Cohort of Sri Lankan Patients with Thromboembolic Disorders.

Prothrombin (F2) 20210G>A [rs1799963 G>A] mutation is a genetic variant which predisposes to inherited thrombophilia. Highest prevalence of this rare mutation has been reported among Caucasian and Mediterranean populations with thrombophilic conditions compared to healthy controls. It is absent or occurs in a very low frequency in both thrombophilic patients and healthy controls of most South Asian populations. A previous study has demonstrated that the mutant allele is absent among Sri Lankan healthy controls. This study was conducted to determine the prevalence of the F2 20210G>A mutation among Sri Lankan patients with thrombo-embolic disorders. F2 20210G>A mutation analysis was carried out on 825 patients. These included 374 with arterial thromboembolic disorders, 303 with venous thromboembolic disorders (VTE) and 148 with pregnancy related complications. Genotyping was done using polymerase chain reaction followed by restriction fragment length polymorphism. The overall prevalence of the individuals detected with the mutation was 0.8% (7/825) with a mutant allele frequency of 0.4% (7/1,650), and all were heterozygotes. Further classification according to the types of thrombotic events showed a prevalence of 0.5% (2/374), 1.3% (4/303), and 0.7% (1/148) respectively, in the three groups with arterial thrombosis, VTE and pregnancy complications. The respective mutant allele frequencies in the three different groups were 0.3% (2/748), 0.7% (4/606) and 0.3% (1/296). Although these figures are lower than that of Caucasian and Mediterranean populations, they are relatively higher compared to other South Asian populations. Therefore, the F2 20210G>A mutant allele is not entirely absent among Sri Lankan patients with thrombo-embolic disorders.

Drug-induced gingival overgrowth: the genetic dimension.

Background:Currently, the etiology of drug-induced gingival overgrowth is not entirely understood but is clearly multifactorial. Phenytoin, one of the common drugs implicated in gingival enlargement, is metabolized mainly by cytochrome P450 (CYP)2C9 and partly by CYP2C19. The CYP2C9 and CYP2C19 genes are polymorphically expressed and most of the variants result in decreased metabolism of the respective substrates.Aims:The present study was undertaken to investigate the influence of the CYP2C9*2 and *3 variant genotypes on phenytoin hydroxylation in subjects diagnosed with epilepsy from South India, thus establishing the genetic polymorphisms leading to its defective hydroxylation process.Materials and Methods:Fifteen epileptic subjects, age 9 to 60 years were included in the study. Among the study subjects, 8 were males and 7 were females. Genomic DNA was extracted from patients’ blood using Phenol-chloroform method and genotyping was done for CYP2C9 using customized TaqMan genotyping assays on a real time thermocycler, by allelic discrimination method. The genetic polymorphisms *1, *2 and *3 on CYP2C9 were selected based on their function and respective allele frequencies in Asian subcontinent among the Asian populations.Results:CYP2C9*1*2 and CYP2C9*3/*3 were identified with equal frequency in the study population. There were seven subjects with CYP2C9*1/*2 genotype (heterozygous mutant), one subject with CYP2C9*1/*1 (wild type) and seven study subjects with CYP2C9*3/*3 (homozygous mutant).Conclusion:The results obtained in the present study will be helpful in the medical prescription purposes of phenytoin, and a more personalized patient approach with its administration can be advocated.

Biochemical characterization of indigenous Fulani and Yoruba ecotypes chicken of Nigeria

The study was carried out to characterize two indigenous chickens of Nigeria using protein markers; haemolglobin (HB) and carbonic anhydrase (CA). Separation of the two proteins was achieved by cellulose acetate electrophoresis and direct gene counting method was employed to interpret the result. Palentological statistics was used to generate dendrogram that measured genetic similarity within and between each of the population studied. HB was interpreted into three phenotypes: AA, AB and BB which were genetically controlled two codominant alleles HBA and HBB. Allele frequencies of HBA and HBB in Yoruba ecotype chicken were 0.34 and 0.66, respectively, while those of Fulani ecotype were 0.28 and 0.72, respectively. CA was also interpreted into three phenotypes (FF, FS and SS) which are genetically controlled by two codominant alleles CAF and CAS; their respective allele frequencies were 0.33 and 0.67 in Yoruba ecotype chicken and 0.24 and 0.76 in Fulani ecotype chicken. Genetic similarity within ecotype indicated 60% in Fulani, 80% in Yoruba and 40% between Yoruba and Fulani at HB locus while at CA locus, genetic similarity was 69% in Fulani ecotype, 50% in Yoruba ecotype and 42% between Yoruba and Fulani ecotype. Cavalli-Sforza genetic distance between the two Ecotypes was 2.1x10-2. Conclusively, the two populations were genetically related and further studies should focus on other protein markers and at molecular level. Key words: Ecotype, protein marker, genetic distance and selection.

Association between MT-CO3 haplotypes and high-altitude adaptation in Tibetan chicken

Genetic mutation in cytochrome c oxidase subunit III gene (MT-CO3) could influence the kinetics of cytochrome c oxidase (COX), which catalyzes oxygen transport capacity in oxidative phosphorylation. However, the potential relationship between MT-CO3 variants and high-altitude adaptation remains poorly understood in Tibetan chicken. Here, we sequenced MT-CO3 gene of 125 Tibetan chickens and 144 Chinese domestic chickens in areas at a low elevation (below 1000m). Eight single nucleotide polymorphisms (SNPs) were detected; and five of them (m.10081A>G, m.10115G>A, m.10270G>A, m.10336A>G and m.10447C>T) shared by Tibetan chicken and lowland chicken with the significant difference in their respective allele frequencies. Nine haplotypes (H1–H9) were finally defined. Among them, haplotype H4 was positively associated with high-altitude adaptation whereas haplotypes H6, H7 and H8 had negative association with high-altitude adaptation. The Median-joining profile suggested that haplotype H5 had the ancestral position to the other haplotypes but had no significant relationship with high-altitude adaptation. However, there was only m.10081A>G mutation differed from haplotype H4 and H5. Results also suggested that chickens with A allele at m.10081A>G, had over 2.6 times than those with G allele in the probability of the ability to adapt hypoxia. It suggests that the synonymous mutation m.10081A>G may be a prerequisite for shaping high-altitude adaptation-specific haplotypes.

Human leukocyte antigen‐A, ‐B, ‐C, ‐DRB1 and ‐DQB1 allele and haplotype frequencies in an Albanian population from Kosovo

HLA-A, HLA-B, HLA-C, HLA-DRB1 and HLA-DQB1 genotyping was performed in a sample of Albanian population from Kosovo. The comparison of the respective allele frequencies through Fst analysis resulted in a close relationship with the Albanians from Albania, the Bulgarians, FYROM Macedonians and Greeks, while the other neighbouring populations are slightly more distant.

Frequencies of the M918I mutation in the sodium channel of the diamondback moth in China, Thailand and Japan and its association with pyrethroid resistance

In addition to the allele frequencies of the L1014F and T929I mutations which are involved in nerve-insensitive resistance to a pyrethroid, those of the M918I mutation were examined using field strains obtained in China, Thailand, and Japan during 2009–2011. Results show that the resistance allele frequencies at the L1014F site were 89–100%, 97–100% and 65–85%, respectively, for strains in China, Thailand, and Japan. The respective allele frequencies at the T929I site were 86–100%, 70–97% and 58–84% for Chinese, Thai, and Japanese strains. With low frequencies up to 27%, M918I was found in Japan and China, but not in Thailand. The strain homozygous for the M918I and L1014F mutations was established and its resistance level to a pyrethroid was examined. The strain lacks a portion of the sodium channel gene corresponding to the 3′ portion of exon 18a, intron 18, and the 5′ portion of exon 18b. Nevertheless, the strain showed a similar level of resistance to that which was homozygous for the T929I and L1014F mutations.