Effects of Origanum majorana essential oil and antibiotics on the quality of frozen thawed Beni Arouss buck semen.

This study aims to investigate the effects of Origanum majorana (OM) essential oil (EO) at different concentrations and antibiotics on post-thawed Beni Arouss buck semen quality. Semen collection was performed using artificial vagina from eight Beni Arouss bucks. Ejaculates were pooled, divided into 12 equal aliquots, washed and diluted to 400 × 106 sperm/ml (with 7% of glycerol). Skim milk-based extender was supplemented with different concentrations of OM EO (0%; 0.01%; 0.02%; 0.03%; 0.04% and 0.05%) without antibiotics, marked as (CTR-), (M1-), (M2-), (M3-), (M4-) and (M5-), and with 50 mg of streptomycin and 50,000 IU of penicillin per 100 ml, marked as (CTR+), (M1+), (M2+), (M3+), (M4+) and (M5+), respectively. Aliquots were cooled to 4°C, then frozen in 0.25 ml straws with a programmable freezer and finally stored in liquid nitrogen for 48 h. Thawing was performed at 37°C for 30 s. Motility, live sperm, sperm abnormalities, membrane integrity, lipid peroxidation and bacterial growth were evaluated after thawing. Among the tested extenders, M2+ improved all semen quality parameters. Sperm motility, live sperm and membrane integrity increased significantly, while the number of abnormal sperm and bacterial growth decreased significantly. The toxic effect of OM EO, with and without antibiotics, appeared beyond 0.03%. In conclusion, M2+ is recommended to improve the cryopreservation of Beni Arouss buck semen.

Phenotypic and genetic characterization of rhizobia isolated from Hedysarum flexuosum in Northwest region of Morocco.

Seventy bacterial strains were isolated from root nodules of the legume Hedysarum flexuosum grown wild in soils from Northwest Morocco. Repetitive extragenic palindromic (REP)-polymerase chain reaction (PCR) clustered the strains into 30 REP-PCR groups. The nearly complete sequence of the 16S rRNA gene from a representative strain of each REP-PCR pattern showed that 17 strains were closely related to members of the genus Rhizobium of the family Rhizobiaceae of the Alphaproteobacteria. Pairwise alignments between globally aligned sequences of the 16S rRNA gene indicated that the strains from H. flexuosum had 99.75-100% identity with Rhizobium sullae type strain IS123(T). The phenotypic characteristics analyzed allowed description of a wide physiological diversity among the isolates, where the carbohydrate assimilation test was the most discriminating. Analysis of the 16S rRNA gene of a representative strains from the remaining 13 REP-PCR groups showed they belong to a wide variety of phylogenetic groups being closely related to species of genera Stenotrophomonas, Serratia, Massilia, Acinetobacter, Achromobacter, and Pseudomonas from the Beta- and Gammaproteobacteria. The R. sullae strains identified in this study produced effective symbiosis with their original host plant. None of the other bacterial strains could form nodules on H. flexuosum.

Quality of life assessment of breast cancer survivors in Northern Morocco: Rural-urban disparity.

This article is a cross-sectional study of 216 women undergoing adjuvant hormone therapy for breast cancer in two oncology centers in northern Morocco. Quality of life (QoL) was assessed using the Functional Assessment of Cancer Therapy (FACT) questionnaire and its endocrine subscale (ES). The relationship between rural-urban status in our sample and QoL was assessed by linear regression analysis using sociodemographic and clinical variables as covariates. Our results show that physical and functional well-being are significantly (p < 0.001) higher in rural areas (24 and 29, respectively) than in urban areas (16 and 19, respectively), while social and emotional well-being are significantly (p < 0.001) higher in urban areas (22 and 21, respectively) than in rural areas (15 and 16, respectively). However, there was no significant difference (p = 0.097) between rural and urban breast cancer survivors regarding endocrine symptom burden. Regarding the effect of sociodemographic and clinical factors on overall HRQOL of breast cancer survivors, hormone type was shown to have a significant effect on overall HRQOL (FACT-ES) of rural and urban breast cancer survivors (𝛽 = +0.849 and 𝛽 = +0.678, respectively). A similar effect was observed for ES (𝛽 = +0.896 and 𝛽 = +0.180, respectively).In contrast, other factors (age, marital status, economic status, menopausal status, type of surgery) did not have a significant effect on HRQOL (FACT-ES) or ES.The study highlighted the need for increased psychosocial supportive care efforts for rural breast cancer survivors to improve their QoL.

Gamma irradiation-induced genetic variability and its effects on the phenotypic and agronomic traits of groundnut (Arachis hypogaeaL.).

In order to increase genetic variability for the improvement of groundnut, two varieties, namely Kp29 and Fleur11, were treated with six different gamma irradiation doses. A significant effect of mutagenesis was distinctly observed in the stem lengths, roots, and survival percentage in both varieties. The radio-sensitivity test showed a mean lethal dose of 436.51Gy for Kp29 and 501.18 Gy for Fleur11. Furthermore, this study revealed putative mutants with variable agro-morphological traits. Seven chlorophyll mutants and various seed shape and color mutants were obtained. This study demonstrates the potency of gamma irradiation to induce high genetic variability that led to the emergence of certain mutations of economic importance.

Bradyrhizobium cytisi sp. nov., isolated from effective nodules of Cytisus villosus.

Several strains isolated from Cytisus villosus nodules have been characterized based on their diverse genetic, phenotypic and symbiotic characteristics. According to 16S rRNA gene sequence analysis, the isolates formed a group that was closely related to Bradyrhizobium canariense BTA-1(T) with 99.4% similarity. Analysis of three housekeeping genes, recA, atpD and glnII, suggested that the C. villosus strains represent a novel Bradyrhizobium species most closely related to B. canariense BTA-1(T) with similarities of 94.2, 96.7 and 94.5%, respectively. All these differences were congruent with DNA-DNA hybridization analysis, which revealed 31% relatedness between a representative strain (CTAW11(T)) isolated from C. villosus nodules and B. canariense BTA-1(T). Phenotypic differences among the strains isolated from C. villosus and B. canariense were based on assimilation of carbon and nitrogen sources. The nodC and nifH genes of strain CTAW11(T) were phylogenetically related to those of strains belonging to bv. genistearum and divergent from those of bv. glycinearum and, accordingly, they do not nodulate soybean. Based on the genotypic and phenotypic data obtained in this study, our strains should be classified as representatives of a novel species for which the name Bradyrhizobium cytisi sp. nov. is proposed; the type strain is CTAW11(T) (=LMG 25866(T)=CECT 7749(T)).

Phenotypic and genotypic characterizations of rhizobia isolated from root nodules of peanut (Arachis hypogaea L.) grown in Moroccan soils.

The phenotypic and genotypic characterization of sixty-two rhizobial isolates obtained from nodules of Arachis hypogaea in north-western Morocco was performed. Their physiological and biochemical properties revealed a great deal of diversity among them. Isolates were classified into two major groups based on the numerical analysis of their phenotypic and genotypic characteristics. Isolates in the first group were alkali- and salt-sensitive, slow or extra-slow growers; they did not use disaccharides as carbon source and varied in the use of amino acids. ARDRA analysis of the 16S rDNA region grouped them together with reference strains belonging to the genus Bradyrhizobium. In the second group, isolates were fast growers, acid-sensitive, and alkali- and salt-tolerant; they used both mono and disaccharides as carbon sources, and methionine was the only amino acid they could metabolize as a nitrogen source. ARDRA analysis grouped them with fast-growing reference strains. Both groups exhibited a range of variability in tolerance to heavy metals. The Intergenic Spacer (IGS)-PCR fingerprinting analysis confirmed a high genotypic diversity at the strain level. This characterization provides a basis for the selection of peanut-nodulating rhizobia which may have applications in formulating appropriate inocula for improving peanut crop yield on Moroccan soils, including saline and acidic marginal areas.

Prevalence of PMQR genes in E. coli and Klebsiella spp. isolated from North-West of Morocco.

OBJECTIVES: Plasmid mediated quinolone resistance (PMQR) genes are increasingly detected worldwide. However, in Morocco a few studies have been carried out. The aim of this study was to investigate the prevalence of these determinants among Escherichia coli and Klebsiella spp. isolated from North-West of Morocco. METHODS: The prevalence of PMQR genes was investigated among 398 E. coli and 118 Klebsiella spp. clinical isolates collected between 2012 and 2015 from North-West of Morocco. Antimicrobial susceptibility was determined by disc diffusion assay and minimal inhibitory concentrations (MIC) of quinolone were determined by micro-dilution. The screening of qnrA, qnrB, qnrC qnrS, Aac(6')-Ib and qepA genes were done by PCR, DNA sequencing and RFLP analyses. RESULTS: Among 398 E. coli and 118 Klebsiella spp. analyzed, 51% of E. coli and 61% of Klebsiella spp. were multidrug resistant (MDR). For the E. coli group, qnrB, qnrS, and aac(6')-Ib-cr genes were detected in 05.02%, 01.50%, and 13.81%, respectively. These determinants were more prevalent in Klebsiella spp. group, hence they represented 11.86%, 05.93% and 18.64% respectively. QnrA, qnrC and qepA were absent in this study. For aac(6')Ib determinant, 84.41% were belong to the Aac(6') Ib-cr variant while for qnrB and qnrS the most determined variants were qnrB1, B6, B16, B42, B66 and qnrS1, S4, S7. Different plasmid sizes were detected in PMQR strains. CONCLUSION: This is the first study conducted in North-West of Morocco and shows important dissemination of MDR and PMQR among Enterobacteriaceae.

[Effectiveness of aqueous extracts of aromatic and medicinal plants against tomato grey mould in Morocco]. / Efficacité des extraits aqueux des plantes aromatiques et médicinales contre la pourriture grise de la tomate au Maroc.

Grey mould is a major disease threatening the Moroccan tomato; this disease is often controlled by fungicides. However, the latter are a real danger to human health and environment. Thus, this study is part of the research of harmless alternatives such extracts of aromatic and medicinal plants (Lavandula officinalis, Thymus vulgaris, Cymbopogon citratus, and Melissa officinalis). In this study, the extracts of four medicinal and aromatic plants were tested for their antifungal potency in vitro and in vivo in order to select the most effective. The results show that, in vitro, the Lavandula officinalis, Thymus vulgaris and Cymbopogon citratus aqueous extracts all possess significant antifungal activity, whereas Melissa officinalis shows the least effective. Also in vivo only the aqueous extract of Cymbopogon citratus proves most effective against B. cinerea on tomato fruit. The test of the plants confirms that aqueous extracts of Cymbopogon citratus and Thymus vulgaris are most effective, while the aqueous extracts of Melissa officinalis and Lavandula officinalis always seem to be the least effective. Therefore, the aqueous extracts of Cymbopogon citratus and Thymus vulgaris are the most envisaged for the biological control of grey mould.

Bradyrhizobium rifense sp. nov. isolated from effective nodules of Cytisus villosus grown in the Moroccan Rif.

Two bradyrhizobial strains, CTAW71(T) and CTAW69, previously isolated from root nodules of Cytisus villosus, have been analysed using a polyphasic approach. These strains have identical 16S rRNA genes and their closest relative species is Bradyrhizobium cytisi, whose type strain CTAW11(T) presented 99.8% identity with respect to strain CTAW71(T). Despite the closeness of the 16S rRNA genes, the housekeeping genes recA, atpD and glnII harboured by strain CTAW71(T) were divergent to those from B. cytisi CTAW11(T), with identity values of 93%, 95% and 97%, respectively. These differences were congruent with DNA-DNA hybridization analysis that revealed an average of 37% relatedness between strain CTAW71(T) and B. cytisi CTAW11(T). Phenotypic characteristics were identical for strains CTAW71(T) and CTAW69, but differed from those of the described species from genus Bradyrhizobium. Based on the genotypic and phenotypic data obtained in this study, we propose that strains CTAW71(T) and CTAW69 should be classified into a new species for which the name Bradyrhizobium rifense sp. nov. is proposed (type strain CTAW71(T)=LMG 26781(T)=CECT 8066(T)).

Characterization of Bradyrhizobium species isolated from root nodules of Cytisus villosus grown in Morocco.

From a total of 73 bacterial strains isolated from root nodules of Cytisus villosus grown in soils of the central-western region of the Moroccan Rif, 68 strains clustered into 19 repetitive extragenic palindromic (REP)-polymerase chain reaction (PCR) groups. The nearly complete 16S rRNA gene sequence from each strain showed they were closely related to members of the genus Bradyrhizobium of the Alphaproteobacteria, but affiliation at the species level was not clear. Sequencing of the housekeeping genes glnII and recA, and their concatenated phylogenetic analysis showed that 11 out of the 19 strains belong to Bradyrhizobium canariense and that another three strains were Bradyrhizobium japonicum. The remaining five strains represented new lineages within the genus Bradyrhizobium since they were not identified with any previously described species. Sequencing of the symbiotic nodC and nifH genes from each bradyrhizobial strain revealed they were all similar to those of the strains included in biovar genistearum.

Characterization of Salmonella Enteritidis isolated from foods and patients in northern Morocco.

OBJECTIVES: This study aimed to investigate Salmonella Enteritidis strains isolated from human and food sources in the north of Morocco by means of phenotypic and genotypic methods. METHODOLOGY: Fifteen isolates from humans and food were submitted to phage typing, XbaI-macrorestriction (pulsed field gel electrophoresis [PFGE]), enterobacterial repetitive intergenic consensus (ERIC-PCR), antimicrobial susceptibility testing, and PCR assay targeting the spvR and invA genes. RESULTS: Six fingerprinting profiles were obtained with the ERIC-PCR method, four with PFGE profiling, five with antimicrobial resistance, three with phage typing, and only one with plasmid profiling. spvR gene was detected in six strains, which did not harbour plasmids of 90 kb. CONCLUSIONS: The conclusions of this study are drawn from a limited number of isolates. It would be desirable to investigate a greater and more diverse population of Salmonella isolates. S. Enteritidis was genotyped and showed four different patterns by PFGE and six by ERIC-PCR. Accordingly, high genetic similarity and limited genetic diversity were found for these strains from north of Morocco.

Isolation, drug resistance and molecular characterization of Salmonella isolates in northern Morocco.

OBJECTIVES: The objective of this investigation was to assess the methods for the characterization of Salmonella isolates and to identify relationships of Salmonella isolates from human and food sources in northern Morocco. METHODOLOGY: Several Salmonella serotypes were isolated from human and food samples and were characterized using conventional culture methods, biochemical, serological, antimicrobial testing, and phage typing. Molecular analyses such as enterobacterial repetitive intergenic consensus (ERIC)-PCR, macrorestriction profiling by pulsed-field gel electrophoresis (PFGE), and virulence gene analysis were also performed. RESULTS: Sixteen Salmonella strains were isolated in our laboratory, serotyped and identified as S. Kottbus, S. Indiana, S. London, S. Typhi, S. Hadar, S. Corvallis, S. Mbandaka, S. Ouakam, S. Tm var. cop., S. Virchow, and S. Altona. The most common resistance profiles for the isolates was ATCFATSCGKSSS, belonging to phage type PT20, ATASCSS associated with strains DT104L/ad and ATATSS for isolates that were not typeable. The PFGE patterns were different for each Salmonella serotype. All strains were negative for the virulence gene spvR. CONCLUSIONS: To our knowledge, this is the first molecular characterization of Salmonella in food and humans from Morocco. Comparison of molecular techniques for differentiating between human and food isolates of Salmonella in north of Morocco shows that ERIC typing and PFGE were more discriminating than the other techniques used in this study.