Mintage Journal of Pharmaceutical & Medical Sciences

Vol 6, Suppl 2, 2017 ISSN: 2320-3315

Original Article

DISTRIBUTION OF ABO AND RH BLOOD GROUPS IN THE REGION OF

ASMAE BENALLA1,2*, AHMED KARIM2, NABIHA TROUGOUTY2, ZAINA SIDQI2, HASSANE MEKHFI1 & MOHAMMED BENAJIBA3 1Laboratoire de Physiologie et Ethnopharmacologie, URAC 40, Faculté des Sciences, Université Mohammed Ier, BP 717 60000, , Morocco, 2Centre Régional de Transfusion Sanguine, Rue Errazi, Oujda, Morocco, 3Centre National de Transfusion Sanguine et d’Hématologie 472, rue Mfadel Cherkaoui, q. Al Irfane, Rabat, Email: [email protected] Received - 02.04.2017; Reviewed and accepted -22.04.2017

ABSTRACT Objective: The aim of this study is to determine the distribution of phenotypic and allelic frequencies of blood groups ABO and RH in the provinces of the Oriental region of Morocco. Methodology: ABO and RH grouping were performed on 3289 blood donors from the seven provinces Berkane, Driouech, , , , Oujda and Taourirt. Results: Phenotypic frequencies O, A, B and AB were respectively 45.36%, 33.75%, 16.10% and 4.79%. Allelic frequencies O, A and B were respectively 0.673, 0.216 and 0.111. In the RH system, DCcee phenotype was the most common (39.27%), followed by DCCee and Dccee (≈16%), DccEe, ddccee and DCcEe (less than 11%), then DccEE with a frequency less than 3%. The other phenotypes were rare. Conclusion: This study will provide in the important database of blood groups ABO and RH for regional blood banks, especially rare phenotypes and for researchers in genetic populations to explain variations between frequencies in provinces. Keywords: ABO blood groups, RH blood groups, phenotypic frequencies, allelic frequencies, Oriental region of Morocco. INTRODUCTION The Oriental region is located on the eastern border of Morocco regional blood bank at Oujda city, or at blood mobile collections in Northern Africa. Given its strategic location, the country has in the seven provinces. been in its history controlled by different populations. Originally inhabited by Berbers considered as the indigenous population, Blood grouping ABO and RH systems have been performed on Oriental region has been then controlled successively by the microplate by the manual method, with monoclonal antisera. Phoenicians, the Romans, the Vandals and the Byzantines. In Data entry and calculations were performed on Microsoft Office the 7th century, Arabic peoples entered the region and their Excel. Allelic frequencies O, A and B were determined by the civilization thrived there. method of Bernstein [8]. Haplotype frequencies were calculated by the formulas proposed by Mourant [9]. Comparisons of ABO and RH are the most important blood group antigens. frequencies were performed by the Chi-square test. Their distribution has been investigated in several populations in the world since their discovery, which shows considerable RESULTS variations between regions [1, 2, 3]. In our study, blood type O was prevalent in the total sample Few studies exist on blood groups in Morocco, some were (45.36%), followed by A (33.75%), B (16.10%) and AB (4.79%). performed on blood donors for ABO and/or RHD [4, 5], others In the provinces, blood type O was always prevalent with a were limited to specific populations, such as the Berber Middle maximum frequency at Taourirt (50.47%), but in Nador, its Atlas [6] and Arabic population of Bni Mellal [7]. frequency was lower to that of blood type A, respectively 36.26% and 36.84% (Table 1). Blood type A was prevalent in The present study, performed for the first time, aims to the north including Nador and Driouech provinces. Comparison investigate the distribution of phenotype and allele frequencies of phenotypic frequencies of ABO blood groups in provinces of ABO and RH (D, C , E, c, e) in provinces of the Oriental shows a highly significant difference (p = 0.0055; χ2 = 36.82; df region of Morocco. The data obtained will be helpful to the blood = 18). management of regional blood bank and will contribute to develop a database of rare erythrocyte phenotypes. This study Allele O was the most prevalent (0.673), followed by A (0.216) will also be helpful to researchers of population genetics to and B (0.111). Allele O was highly frequent in Taourirt (0.710), understand the migration of populations in this region. whereas, the lowest frequency was observed in Nador (0.602). Allele A frequency was between 0.191 at Jerada and 0.260 at METHODOLOGY Nador. Allele B frequency was between (0.091) at Driouech and The Oriental region of Morocco is one of 16 regions from the 0.145 at Nador (Table 1). administrative division of Morocco. This region, with an area of In the oriental region, the increasing order of distribution of ABO 82820 Km², includes 7 provinces: Berkane, Driouech, Figuig, RHD phenotypic frequencies was: Opos > Apos > Bpos > Jerada, Nador, Oujda and Taourirt. ABpos > Oneg > Aneg > Bneg > ABneg (Table 2). This order The Oriental region is characterized by its strategic position was observed in all provinces except in Nador where Apos was between Algerian borders at east, Mediterranean coast, close to more frequent. Concerning groups RHD negatif, Oneg is the the southern of Europe at the north and, near north of Sahara at most frequent in Figuig (6.25%), whereas Bneg has the higher the south. frequency in Nador (2.05%). Aneg is more frequent in Berkane and Taourirt, respectively 4.41% and 3.56%. The sample consists of 3298 voluntary blood donors of both sexes, aged 18-60 years, living in the region. They were Concerning RH system, the major phenotypes found in our collected between July 2010 and May 2013. Each donor was study were DCcee (39.27%), followed by DCCee and Dccee counted only once. Blood samples were collected in either (respectively 16.19% and 16.07%), DccEe (9.85%), ddccee

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(8.73%), DCcEe (7.28%), and DccEE (1.49%) (Table 3). The In RH haplotypes, the distribution pattern was: Other phenotypes (DCCEe, DCcEE, ddCcee, ddCCee and CDe>cde>cDe>cDE>Cde>cdE>CDE (Table 4). Some ddccEe) are rare, less than 1%. The higher frequency of DCCee differences are observed between provinces, thus, cde and cDe was observed in Jerada (20.41%), when Dccee and ddccee are more prevalent than CDe in Figuig with the same frequency were more prevalent in Figuig (respectively 20.63% and (2.2932). In Driouech, the haplotype cDe is more prevalent than 10.31%). Comparing frequencies of the major six phenotypes in cde (respectively 0.3114 and 0.2264). the provinces shows a significant difference (p = 0.045; χ2 =44.18; df=30). Table 1: Phenotypic (%) and allelic frequencies of ABO system in provinces of Oriental region of Morocco

Provinces Sample Phenotypic frequencies Allelic frequencies O A B AB O A B N % N % N % N % Berkane 884 392 44.34 301 34.05 141 15.95 50 5.66 0.6659 0.2235 0.1146 Driouech 132 61 46.21 48 36.36 18 13.64 5 3.79 0.6798 0.2264 0.0913 Figuig 320 140 43.75 111 34.69 50 15.63 19 5.94 0.6614 0.2294 0.1144 Jerada 387 180 46.51 123 31.78 73 18.86 11 2.84 0.6820 0.1915 0.1152 Nador 342 124 36.26 126 36.84 63 18.42 29 8.48 0.6021 0.2606 0.1450 Oujda 700 330 47.14 237 33.86 109 15.57 24 3.43 0.6866 0.2081 0.1000 Taourirt 533 269 50.47 167 31.33 77 14.45 20 3.75 0.7104 0.1943 0.0956 TOTAL 3298 1496 45.36 1113 33.75 531 16.10 158 4.79 0.6735 0.2160 0.1106

Table 2: Phenotypic (%) frequencies of ABO and RHD groups in provinces of Oriental region of Morocco.

Provinces Sample O A B AB RHDpos RHDneg RHDpos RHDneg RHDpos RHDneg RHDpos RHDneg N % N % N % N % N % N % N % N % Berkane 884 358 40.50 34 3.85 262 29.64 39 4.41 127 14.37 14 1.58 45 5.09 5 0.57 Driouech 132 55 41.67 6 4.55 47 35.61 1 0.76 17 12.88 1 0.76 4 3.03 1 0.76 Figuig 320 120 37.50 20 6.25 99 30.94 12 3.75 45 14.06 5 1.56 17 5.31 2 0.63 Jerada 387 160 41.34 20 5.17 108 27.91 15 3.88 69 17.83 4 1.03 8 2.07 3 0.78 Nador 342 110 32.16 14 4.09 117 34.21 9 2.63 56 16.37 7 2.05 24 7.02 5 1.46 Oujda 700 304 43.43 26 3.71 221 31.57 16 2.29 102 14.57 7 1.00 23 3.29 1 0.14 Taourirt 533 252 47.28 17 3.19 148 27.77 19 3.56 70 13.13 7 1.31 17 3.19 3 0.56 TOTAL 3298 1359 41.21 137 4.15 1002 30.38 111 3.37 486 14.74 45 1.36 138 4.18 20 0.61

Table 3: RH phenotypic frequencies (%) in provinces of the Oriental region of Morocco.

Provinces Sample DCcee DCcEe DCCee DccEe DccEE Dccee DCCEe DCcEE ddccee ddCcee ddCCee ddccEe Berkane 884 38.01 7.24 14.48 10.86 1.70 17.19 0.00 0.11 10.07 0.23 0.00 0.11 Driouech 132 37.88 9.09 14.39 13.64 1.52 16.67 0.00 0.00 6.06 0.00 0.76 0.00 Figuig 320 40.63 7.19 11.56 6.56 1.25 20.63 0.00 0.00 10.31 1.88 0.00 0.00 Jerada 387 38.24 7.75 20.41 8.27 1.03 12.14 1.03 0.26 10.08 0.52 0.00 0.26 Nador 342 36.26 6.73 16.96 11.70 1.46 16.37 0.29 0.00 9.65 0.29 0.00 0.29 Oujda 700 41.29 6.71 18.43 9.57 1.71 14.71 0.43 0.00 6.14 0.43 0.00 0.57 Taourirt 533 40.90 7.69 15.76 9.57 1.31 15.76 0.38 0.00 8.07 0.56 0.00 0.00 TOTAL 3298 39.27 7.28 16.19 9.85 1.49 16.07 0.30 0.06 8.73 0.52 0.03 0.21

Table 4: RH haplotypic frequencies in provinces of Oriental region of Morocco.

Provinces Sample CDE CDe cDE cDe Cde cdE cde Berkane 884 0.0000 0.3430 0.1169 0.2465 0.0032 0.0016 0.2887 Driouech 132 0.0000 0.3490 0.1132 0.3114 0.0000 0.0000 0.2264 Figuig 320 0.0000 0.2849 0.1021 0.2932 0.0267 0.0000 0.2932 Jerada 387 0.0064 0.4150 0.0914 0.1789 0.0076 0.0038 0.2969 Nador 342 0.0018 0.3671 0.1049 0.2376 0.0042 0.0042 0.2801 Oujda 700 0.0029 0.3795 0.1082 0.2677 0.0078 0.0104 0.2235 Taourirt 533 0.0026 0.3599 0.1065 0.2578 0.0092 0.0000 0.2640 TOTAL 3298 0.0020 0.3599 0.1082 0.2485 0.0080 0.0033 0.2701

DISCUSSION The A and B allele frequencies show a north-south gradient from North Mediterranean countries to sub-Saharan Africa, and Blood group determination has a great importance and is used east-west gradient, from North Africa to the Middle East and in several biological disciplines, such as population genetics [1, other Asia countries. Thus, the A allele frequency increases 2], human evolution and medicine [10]. So, each blood bank from south to north. This allele is more common in Caucasians, should study blood groups frequencies in their region, especially and it decreases in the direction of the east. The opposite is ABO and RH groups which are the most important. observed for the B allele which is more frequent in Asia and In the Oriental region, blood group O was the more sub-Saharan Africa [1, 13, 14, 15], its frequency decreases predominant, followed by A, Band AB. However, in Northern towards the north. provinces (Nador and Driouech) we can found the higher The RH phenotype frequencies in the Oriental region of frequencies of blood type A. the same results were described in Morocco show that the DCcee phenotype is far the most Morocco by Habti [4] and Benahadi [5] who observed a north- predominant. Some phenotypes RH which have low frequencies south gradient distribution of ABO alleles: the O allele frequency (DCCEe, DCcEE, ddccEe, ddCcee, ddCCee) and phenotypes decrease from south to north of Morocco and that of the A allele not found (DCCEE, ddccEE, ddCCEe, ddCcEE, ddCCEE and increase from south to north. In comparison with neighboring ddCcEe) in our region are generally also rare phenotypes in countries, our results are similar to those reported in North other populations [9, 12, 16]. Africa [11, 12].

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Haplotypes that are well represented in the region are classified Distribution of ABO and Rhesus D blood antigens in as follows according to their frequencies: CDe>cde>cDe>cDE. Morocco. Internet J Biol Anthropol 2013; 6 (1): 1–6. The frequencies of these haplotypes could be insertedamong 6. Harich N, Esteban E, Chafik A, Lopez-Alomar A, Vona G those found in North Africa [17, 18]. Haplotype frequencies and Moral P. Classical polymorphisms in Berbers from CDe, cde and cDe follow gradients from northern Mediterranean Moyen Atlas (Morocco): Genetics, geography, and countries to sub-Saharan Africa. Thus, CDe and cde decreases historical evidence in the Mediterranean peoples. Ann Hum from north to south, while cDe increases from north to south, Biol 2002; 29(5): 473–87. [19, 20, 21, 22]. 7. El Ossmani H, Bouchrif B, Glouib K, Zaoui D, El Amri H and Chafik A. Etude du polymorphisme des groupes ABO RH frequencies observed in the Oriental region of sanguins (ABO, Ss, RHESUS ET DUFFY) chez la Morocco, which are intermediate between those of sub-Saharan population arabophone du plateau de Beni Mellal. Leb Sci Africa in south and East Asia and those of northern J 2008; 9(1): 17–28. Mediterranean suggest that our region, by its location, has been 8. Nan Juan MO and Gart JJ. Bernstein’s and gene counting influenced by populations of the three regions in its history. methods in generalized ABO like systems. Ann Hum Genet Indeed, this region has been in the history controlled by different London 1976; 39: 361–73. populations. First inhabited by Berbers, considered the 9. Mourant AE, Koipec A and Domaniewska-Sobczak K. The indigenous people, and then by the Romans, the Vandals, the Distribution of the Human Blood Groups and other Byzantines and the Arabs. Therefore, the immuno- Polymorphisms. Oxford University Press London, 1976. hematological profile of the region had to undergo all these 10. Peyrard T, Pham BN, Le Pennec PY and Rouger P. Les influences that have contributed to the creation of an phénotypes érythrocytaires rares: un enjeu de santé intermediate phenotype. publique. Transfus Clin Biol 2008; 15: 109–119. CONCLUSION 11. Aireche H and Benabadji M. Les fréquences géniques dans les systèmes ABO, P et Luthéran en Algérie. This study is the first that documented the distribution of ABO Transfus Clin Biol 1994; 1(4): 279–289. and RH blood groups among provinces of the Oriental region of 12. Said N, Ben Ahmed F, Doghri A, Ghazouani E, Layouni S, Morocco. Frequency distribution of ABO and RH groups in Gritli N and Nsiri B. Polymorphisme ABO dans une provinces of the Oriental region is not uniform, which denotes population de donneurs de sang tunisiens. The ABO the variability of populations living in the region. This distribution system polymorphism in Tunisian blood donors. Transfus could have interesting implications for the blood banks of Clin Biol 2003; 10: 331–334. Oriental since it shows the presence of rare phenotypes and 13. Reid ME and Lomas-Francis C. The Blood Group Antigen allows to highlight sites rich in phenotypes used in emergency Facts Book. 2è ed., New York: Elsevier Academic Press; conditions. This is the case of Group O ddccee which remain 2004. the first choice when vital emergency, as it presents the least 14. Dean L. Blood groups and red cell antigens. National risk. This suggests the need for more blood collections in Center for Biotechnology Information (NCBI), 2005. provinces with a high frequency of this phenotype. 15. Raja KA, Dobariya GH, Unagar CA, Pandya AN, Patel JN and Wadhwani SN. Frequency and distribution of ABO and RECOMMENDATIONS Rh blood groups among blood donors in tertiary care In the absence of Red Blood Cell cryopreservation process and hospital of South Gujarat. India. Int J Res Med Sci 2016; 4: national bank Red Blood Cell rare phenotypes, the existence of 5377–5381. rare phenotypes in our region is a challenge and requires the 16. Benahadi A, Boulahdid S, Adouani B, Laouina A, Mokhtari establishment of a local strategy applicable to the Oriental blood A, Soulaymani A, Hajjout K, Benajiba M and Alami R. banks to provide blood for such patients, particularly multi- Mapping Rare Erythrocyte Phenotypes in Morocco: A Tool transfused. On the other hand, the results of this study would be to Overcome Transfusion Challenges. J Blood Transfus helpful to the researchers in population genetics to explore the 2014; 1–4. factors responsible for the observed distribution of blood groups 17. Aireche H and Benabadji M. 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