High Level of Heterozygous Haplotype of Hemoglobin in Abidjan

Background. Sickle cell disease (SCD) is an hemoglobin disorder that concerns 300,000 newborns each year around the world. Hemoglobin haplotypes can modulate SCD clinical expression. In Côte d’Ivoire, no study has yet investigated the distribution of hemoglobin haplotypes in the population. The goal of this study was to identify hemoglobin haplotypes for people attending dispensary with mild malaria in Abidjan (Ivory Coast) independently of their SCD status. Methods To determine haplotypes, specific restriction enzyme (RE) method is used after PCR amplification with different couples of primers. According to the digestion profile of PCR products, five hemoglobin haplotypes are found over the world. ResultsIn Abidjan, four different “classical” haplotypes of hemoglobin were detected: Benin (56.5%), Bantou (28.5%), Senegal (4%), Cameroun (1%). In parallel 10% of atypical profiles were described. Heterozygous haplotype (69%) was more frequent than homozygous haplotype (31%). ConclusionsBenin haplotype usually associated with more severe of SCD symptoms was predominant in the study population. However this preliminary study highlights also a high prevalence of atypical and heterozygous haplotypes in the population.

Directed Recovery and Molecular Characterization of Antibiotic Resistance Plasmids from Cheese Bacteria

Resistance to antimicrobials is a growing problem of worldwide concern. Plasmids are thought to be major drivers of antibiotic resistance spread. The present work reports a simple way to recover replicative plasmids conferring antibiotic resistance from the bacteria in cheese. Purified plasmid DNA from colonies grown in the presence of tetracycline and erythromycin was introduced into plasmid-free strains of Lactococcus lactis, Lactiplantibacillus plantarum and Lacticaseibacillus casei. Following antibiotic selection, the plasmids from resistant transformants were isolated, analyzed by restriction enzyme digestion, and sequenced. Seven patterns were obtained for the tetracycline-resistant colonies, five from L. lactis, and one each from the lactobacilli strains, as well as a single digestion profile for the erythromycin-resistant transformants obtained in L. lactis. Sequence analysis respectively identified tet(S) and ermB in the tetracycline- and erythromycin-resistance plasmids from L. lactis. No dedicated resistance genes were detected in plasmids conferring tetracycline resistance to L. casei and L. plantarum. The present results highlight the usefulness of the proposed methodology for isolating functional plasmids that confer antibiotic resistance to LAB species, widen our knowledge of antibiotic resistance in the bacteria that inhabit cheese, and emphasize the leading role of plasmids in the spread of resistance genes via the food chain.

A Case Study of the Response of Immunogenic Gluten Peptides to Sourdough Proteolysis

Celiac disease is activated by digestion-resistant gluten peptides that contain immunogenic epitopes. Sourdough fermentation is a potential strategy to reduce the concentration of these peptides within food. However, we currently know little about the effect of partial sourdough fermentation on immunogenic gluten. This study examined the effect of a single sourdough culture (representative of those that the public may consume) on the digestion of immunogenic gluten peptides. Sourdough bread was digested via the INFOGEST protocol. Throughout digestion, quantitative and discovery mass spectrometry were used to model the kinetic release profile of key immunogenic peptides and profile novel peptides, while ELISA probed the gluten’s allergenicity. Macrostructural studies were also undertaken. Sourdough fermentation altered the protein structure, in vitro digestibility, and immunogenic peptide release profile. Interestingly, sourdough fermentation did not decrease the total immunogenic peptide concentration but altered the in vitro digestion profile of select immunogenic peptides. This work demonstrates that partial sourdough fermentation can alter immunogenic gluten digestion, and is the first study to examine the in vitro kinetic profile of immunogenic gluten peptides from sourdough bread.

Differentiation of Two Pleurotus Species Based on the Restrictive Digestion Profile of the Internal Transcribed Spacer Region

Two oyster mushrooms (Pleurotus eous P-31 and P. ostreatus EM-1) are under either cottage industry or semi-commercial cultivation in Ghana. The latter (P. ostreatus) is already well known to the public and on the shelf of some leading supermarkets. There is morphological resemblance between the two species making it difficult for the untrained eye to distinguish between them except for the colour difference. In this study, molecular methods were em­ployed to differentiate among the two species. The Internal Transcribed Spacer ITS 1 and ITS 4 regions of the rDNA of the two oyster species were amplified by the conventional PCR using the universal primer pair, ITS 1 and ITS 4 followed by restrictive digestion with enzymes, (Hh I, Hinf I, Rsa I and Hae III). The two species could not be separated based on the ampli­fied bands only, as both produced a characteristic band size of 650 bp. Gel profiling showing restrictive patterns generated by the four enzymes indicated that only the Hae III restrictive enzyme was effective in separating P. eous P-31 and P. ostreatus EM-1. This is the first record of the separation of the Ghanaian Pleurotus species by molecular methods indicating their genetic differences.

Comparative analysis of IGFBP-3 gene sequence in Egyptian sheep, cattle, and buffalo

Objective: A total of 205 animals from four Egyptian livestock species; cattle (n=18), buffaloes (n=12), sheep (n=150) and goats (n=25) were used in this study to detect polymorphism and perform comparative analysis for IGFBP-3 gene using DNA sequencing and (PCR-RFLP). Results: The amplified fragments were found to be of length 654 bp in sheep, 651bp in cattle and 655 bp in buffalo. For Falahy goats, PCR was performed to amplify a 316 bp fragment from exon 2 of the IGFBP-3 gene. The digestion of 654 bp with HaeIII restriction enzyme yielded a single restriction pattern for goats, while for cattle, 3 genotypes were identified; (AA),(AB), and (BB). Moreover, for buffalo one genotype (AA) only was found with HaeIII and TaqI restriction enzymes, separately. Also, the digestion profile for goats with HaeIII revealed one pattern only. Nucleotide sequencing of the amplified fragments of IGFBP-3 gene in sheep, cattle, buffalo, and goat was submitted to the NCBI GenBank (Accession no. MG738671.1, MG738673.1, MG738674.1, and MG738672.1, respectively). The nucleotide sequencing analysis indicated similarity percentages in IGFBP-3 gene fragments of 88.54, 89.63 and 95.06 % between ''sheep and cattle '', ''sheep and buffalo '', and ''cattle and buffalo'', respectively.

Comparative analysis of IGFBP-3 gene sequence in Egyptian sheep, cattle, and buffalo

Objective: A total of 205 animals from four Egyptian livestock species; cattle (n=18), buffaloes (n=12), sheep (n=150) and goats (n=25) were used in this study to detect polymorphism and perform comparative analysis for IGFBP-3 gene using DNA sequencing and (PCR-RFLP). Results: The amplified fragments were found to be of length 654 bp in sheep, 651bp in cattle and 655 bp in buffalo. For Falahy goats, PCR was performed to amplify a 316 bp fragment from exon 2 of the IGFBP-3 gene. The digestion of 654 bp with HaeIII restriction enzyme yielded a single restriction pattern for goats, while for cattle, 3 genotypes were identified; (AA),(AB), and (BB). Moreover, for buffalo one genotype (AA) only was found with HaeIII and TaqI restriction enzymes, separately. Also, the digestion profile for goats with HaeIII revealed one pattern only. Nucleotide sequencing of the amplified fragments of IGFBP-3 gene in sheep, cattle, buffalo, and goat was submitted to the NCBI GenBank (Accession no. MG738671.1, MG738673.1, MG738674.1, and MG738672.1, respectively). The nucleotide sequencing analysis indicated similarity percentages in IGFBP-3 gene fragments of 88.54, 89.63 and 95.06 % between ''sheep and cattle '', ''sheep and buffalo '', and ''cattle and buffalo'', respectively.

Comparative analysis of IGFBP-3 gene sequence in Egyptian sheep, cattle, and buffalo

Objective: A total of 205 animals from four Egyptian livestock species; cattle (n=18), buffaloes (n=12), sheep (n=150) and goats (n=25) were used in this study to detect polymorphism and perform comparative analysis for IGFBP-3 gene using DNA sequencing and (PCR-RFLP). Results: The amplified fragments were found to be of length 654 bp in sheep, 651bp in cattle and 655 bp in buffalo. For Falahy goats, PCR was performed to amplify a 316 bp fragment from exon 2 of the IGFBP-3 gene. The digestion of 654 bp with HaeIII restriction enzyme yielded a single restriction pattern for goats, while for cattle, 3 genotypes were identified; (AA),(AB), and (BB). Moreover, for buffalo one genotype (AA) only was found with HaeIII and TaqI restriction enzymes, separately. Also, the digestion profile for goats with HaeIII revealed one pattern only. Nucleotide sequencing of the amplified fragments of IGFBP-3 gene in sheep, cattle, buffalo, and goat was submitted to the NCBI GenBank (Accession no. MG738671.1, MG738673.1, MG738674.1, and MG738672.1, respectively). The nucleotide sequencing analysis indicated similarity percentages in IGFBP-3 gene fragments of 88.54, 89.63 and 95.06 % between ''sheep and cattle '', ''sheep and buffalo '', and ''cattle and buffalo'', respectively.

Comparative analysis of IGFBP-3 gene sequence in Egyptian sheep, cattle, and buffalo

Objective A total of 205 animals from four Egyptian livestock species; cattle (n = 18), buffaloes (n = 12), sheep (n = 150) and goats (n = 25) were used in this study to detect polymorphism and perform comparative analysis for IGFBP-3 gene using DNA sequencing and (PCR–RFLP). Results The amplified fragments were found to be of length 654 bp in sheep, 651 bp in cattle and 655 bp in buffalo. For Falahy goats, PCR was performed to amplify a 316 bp fragment from exon 2 of the IGFBP-3 gene. The digestion of 654 bp with HaeIII restriction enzyme yielded a single restriction pattern for goats, while for cattle, 3 genotypes were identified; (AA), (AB), and (BB). Moreover, for buffalo one genotype (AA) only was found with HaeIII and TaqI restriction enzymes, separately. Also, the digestion profile for goats with HaeIII revealed one pattern only. Nucleotide sequencing of the amplified fragments of IGFBP-3 gene in sheep, cattle, buffalo, and goat was submitted to the NCBI GenBank (Accession no. MG738671.1, MG738673.1, MG738674.1, and MG738672.1, respectively). The nucleotide sequencing analysis indicated similarity percentages in IGFBP-3 gene fragments of 88.54, 89.63 and 95.06% between “sheep and cattle”, “sheep and buffalo”, and “cattle and buffalo”, respectively.

Comparative analysis of IGFBP-3 gene sequence in Egyptian sheep, cattle and buffalo

Objective: A total of 205 animals from four Egyptian livestock species were used in this study to detect polymorphism and perform comparative analysis for IGFBP-3 gene using DNA sequencing and (PCR-RFLP). Results: The digestion of 654 bp with HaeIII restriction enzyme yielded a single restriction pattern of eight fragments in sheep revealing the absence of polymorphism, while in cattle 3 genotypes were identified; (AA), (AB), and (BB). Moreover, one genotype (AA) only was found in buffalo using HeaIII and MspI restriction enzymes, separately. Digestion profile for a goat with HaeIII revealed only one pattern for three DNA fragments, which means the absence of polymorphism within the IGFBP-3 gene in the tested goats. The nucleotide sequencing analysis indicated similarity percentages in IGFBP-3 gene fragments of 77, 30 and 58% between ''sheep and cattle '', ''sheep and buffalo '', and ''cattle and buffalo'', respectively.