Molecular detection of antimicrobial resistance genes in multidrug-resistant Gram-negative bacteria isolated from clinical samples in two hospitals in Niger

Background: According to the World Health Organization (WHO), bacterial resistance to antibiotics is a global public health challenge, which is also developing in Niger. The aim of this study was to determine the prevalence of antibiotic resistance genes in Gram-negative bacilli isolated from clinical samples in the biological laboratories of two selected health facilities in Niger. Methodology: Clinical bacterial isolates were randomly collected from two biological laboratories of Zinder National Hospital and Niamey General Reference Hospital. These were multi-resistant Gram-negative bacteria that have been routinely isolated from pathological samples of patients. Molecular detection of resistance genes was carried out by polymerase chain reaction (PCR) amplification using specific primers. These include plasmid-mediated AmpC beta lactamase genes (blaCITM, blaDHAM, blaFOXM), 'Cefotaxime-Munich' type beta lactamase genes (blaCTX-M-1, blaCTX-M-2, blaCTX-M-9), KPC-type beta lactamase gene (blaKPC), Oxa-type beta lactamase gene (blaOXA-48), SHV-type beta lactamase gene (blaSHV), TEM-type beta lactamase gene (blaTEM), quinolone resistance genes (qnrA, qnrB, qnrS), and sulfonamide resistance genes (sul1, sul2, sul3). Results: A total of 24 strains of multidrug-resistant Gram-negative bacteria isolated from different clinical samples were analysed. The distribution of the resistance genes detected is as follows; AmpC blaCITM (n=6; 25.0%), AmpC blaDHAM (n=4; 17.0%), AmpC blaFOXM (n=0), blaCTX-M-1 (n=11; 46.0%), blaCTX-M-2 (n=0), blaCTX-M-9 (n=0), blaKPC (n=0), blaOXA-48 (n=2; 8..0%), blaSHV (n=5; 21.0%), blaTEM (n=0), qnrA (n=0), qnrB (n=5; 21.0%), qnrS (n=17; 71.0%), sul1 (n=22; 92.0%), sul2 (n=12; 50.0%), and sul3 (n=0). All isolates tested had at least two resistance genes. Conclusion: The results of this study provide a better understanding of the resistance situation of clinical isolates in Niger. Therefore, it is more than necessary to intensify the detection on a larger number of samples and on a national scale. This will make it possible to assess the true extent of the phenomenon and consequently guide control strategies through a national multisectoral plan.

Profile of multidrug-resistant clinical bacterial isolates at the National Hospital of Zinder (NHZ), Niger Republic in 2021

Background: Today, bacterial resistance is a public health challenge throughout the world, and infections caused by resistant bacteria are associated with increased morbidity, mortality and health care costs. The objective of this descriptive study is to determine the prevalence and distribution of multi-drug resistant (MDR) clinical bacteria isolates at the National Hospital of Zinder, Niger Republic in 2021. Methodology: We conducted a descriptive cross-sectional study of in- and out-patients from whose clinical samples' bacteria were isolated at the bacteriology unit of the laboratory. Bacteria were isolated from the clinical samples following standard aerobic cultures and identified using conventional biochemical test schemes. Antibiotic susceptibility testing (AST) was performed by the agar disk diffusion technique, and categorization of the isolates into sensitive, intermediate or resistant was done according to the recommendations of the Antibiogram Committee of the French Society of Microbiology (CA-SFM) 2020 version 1.2. MDR was defined as resistance to at least one antibiotic in three or more categories, while selected MDR bacteria such as ESBL was identified using double disk synergy test, and MRSA by cefoxitin disk diffusion test. Results: Seventy-seven (6.7%) bacterial species were isolated from 1153 clinical samples processed at the bacteriology unit of the hospital laboratory between June and December 2021, of which 65.0% (50/77) were members of the order Enterobacteriales. Escherichia coli represented 40.3% (40/77) of the isolated bacteria, Staphylococcus aureus 13.0% (10/77) and Pseudomonas aeruginosa 11.7% (9/77). The overall prevalence of MDR was 44.2% (34/77), including 61.8% (21/34) ESBL-producing Enterobacteriales (ESBL-E), 26.5% (9/34) multi-resistant P. aeruginosa and 11.7% (4/34) MRSA, with 67.6% (23/34) of the MDR isolates from outpatients. Resistance rates of the Enterobacteriales to ciprofloxacin, gentamicin, amikacin and imipenem were 62.0%, 52.0%, 38.0% and 8.0% respectively. Resistance rates of P. aeruginosa were 100.0%, 88.9%, 77.8%, 33.3%, 22.2%, and 22.2% respectively to ceftazidime, ticarcillin, imipenem, ciprofloxacin, levofloxacin, and amikacin. Resistance rates of S. aureus were 100.0%, 50.0%, 40.0%, 10.0%, 0% and 0% to penicillin G,erythromycin, cefoxitin, tetracycline, fusidic acid, and chloramphenicol respectively. ESBL-E were 47.6%,85.7% and 0% resistant to amikacin, ciprofloxacin and imipenem, and MRSA resistance rates were 75.0%, 75.0%, 50.0% and 0% to erythromycin, tetracycline, gentamicin, and chloramphenicol respectively. Conclusion: This study reports high prevalence of MDR bacteria, mainly ESBL-E, with concerning high resistance to carbapenem. Rational use of antibiotics and implementation of surveillance system for MDR bacteria must be implemented in order to limit the emergence and spread of MDR bacteria in Niger Republic.

Preparation of phosvitin-dextran conjugates under high temperature in a liquid system.

The conjugation reaction between phosvitin (Pv) and dextran (Dex) in an aqueous solution at the initial stage of Maillard reaction was investigated in this study. The formation of Pv-Dex conjugates was indirectly traced by measuring the absorbance at 200 nm in the UV-vis spectrophotometer. The content of free amino groups in the glycosylated Pv decreased to 77.4% after reaction for 6 h. The new band near the loading end of SDS-PAGE gel also suggested the high molecular weight products developed after conjugation. The shift of the main peak of Pv in size exclusion-high performance liquid chromatography from 9.16 min to 8.87 min and the minor peak from 6.56 min to 6.13 min confirmed the formation of covalent conjugates. Circular dichroism (CD) spectra demonstrated that significant changes of the secondary structure occurred to the grafted Pv. The optimum conjugation condition chosen was per Pv (5 mg/ml) glycosylated with 4-fold dextran (Mw, 40 kDa) at 100 °C for 6 h.

[Survey on the illicit drug market in Cotonou, Benin in March 2003]. / Enquete sur le marché illicite des médicaments à Cotonou (Bénin) en mars 2003.

Illicit sale of medicines is a serious public health problem in Africa. In Benin, an informational program was undertaken to fight this threat. A three-day survey on consumption of medicines was conducted in various areas in Cotonou. The purpose was to evaluate purchasing practices so that related concerns and beliefs could be used to craft messages for the next campaign to curb illicit medicines. In addition to being a tool for message development, this survey will be used as an evaluation tool to measure message impact during and after the information campaign. A questionnaire designed to evaluate several aspects of drug purchasing behavior was administered by investigators in 600 randomly selected households. The main findings were as follows. Repressive measures to reduce the sale of counterfeit medicines on the illicit market are necessary but inadequate. New campaign messages for targeted groups are necessary since 86% of people interviewed thought that medicines acquired from street vendors were of good quality. More importantly it will be necessary to make legitimate medicines more accessible and affordable (generic drugs) and to simplify dispensation procedures allowing prescriptions to be filled followed by appropriate professional counsel.

[Evaluation of public awareness campaigns on counterfeit medicines in Cotonou, Benin]. / Evaluation des campagnes de sensibilisation sur les pratiques d'achat de médicaments dans la ville de Cotonou.

A public awareness campaign mainly TV and radio announcements to promote public awareness of the dangers of counterfeit medicines was developed based on survey data collected in 2003. The survey was designed to evaluate the purchasing practices of consumers in Cotonou, Benin with regard to medicines. Based on findings, several recommendations were made for crafting new messages about generic drugs and the dangers of the illicit medicines market. Evaluation of the results of the new campaign that lasted 9 months showed that: most respondents stated that the campaign announcements were effective in increasing their awareness; nine out of 10 persons understood the messages on the dangers of the illicit market; a net decrease in buying on the illicit market was observed among respondents that increased their awareness; the frequency of house calls by illicit vendors decreased. Most respondents stated that they requested generic drugs spontaneously. The preferred sources for procurement of generic medicines were public health facilities and pharmacies.

Enquête sur le marché illicite des médicaments à Cotonou (Bénin) en mars 2003

La vente illicite de médicaments est un problème de santé publique en Afrique. Au Bénin, un programme de commu n i c ation pour lutter contre ce fléau a été mis en œuvre. Cette étude sur la consommation des médicaments à Cotonou (Bénin),qui s'est déroulée pendant trois jours dans différents quartiers de la ville, avait pour objet d'évaluer les pratiques d'achat des consommateurs , de façon à tenir compte de leurs préoccupations et de leurs croyances dans l'élab o ration de messages de com- munication pour les prochaines campagnes de lutte contre le marché illicite. Cette enquête est un outil pour la conception des messages et, répétée pendant et après ces campagnes, elle sera aussi un outil d'évaluation de leur impact. Un questionnaire explorant plusieurs thèmes liés à l'ach at de médicaments, a été administré par les enquêteurs au domicile de 600 ménages sélectionnés aléatoirement. Les principaux résultats sont les suivants : les mesures de répression visant à réduire l'offre du mar- ché illicite des médicaments sont nécessaires mais pas suffisantes. Le contenu des messages de sensibilisation devra être revu et réorienté ve rs des groupes cibles , car 86% des personnes interrogées pensent que les médicaments de trottoir sont de bonne qualité. Il faudra surtout augmenter la disponibilité géographique au plus près des populations des médicaments financièrement accessibles (notamment des génériques) et mettre en place des procédures allégées de dispensation et de pre s c ription suivies d'un conseil professionnel approprié

New calibration method for gas chromatographic assay of carbon monoxide in blood.

The calibration method described is based on the in situ formation of carbon monoxide in a closed system by the reaction of hot concentrated sulfuric acid with formic acid. Carbon monoxide is released from hemoglobin by treatment with 85% phosphoric acid, diluted 1:2 (v/v). Carbon monoxide is analyzed by means of headspace gas chromatography on a Porapak Q 80-100 column, following catalytic reduction to methane, using a flame-ionization detector. The method was validated by comparing the results obtained for blood samples from patients suffering from carbon monoxide poisoning and previously analyzed by means of spectrophotometry.