Resistance to protease inhibitors among persons living with HIV in Ghana: a case for viral load and drug resistance monitoring.

OBJECTIVE: Routine viral load and drug resistance testing are well supported in most resource-rich settings and provide valuable benefits in the clinical care of PLWH in these communities. Undoubtedly, there exist financial and political constraints for the scale-up of viral load and drug resistance testing in Sub-Saharan Africa. To achieve the global UNAIDS 95/95/95 targets, there is the need to bridge this inequity in patient care and allow for a universal approach that leaves no community behind. METHODS: Venous blood from 96 PLWH on second-line ART from Korle-Bu Teaching Hospital were collected and processed into plasma for CD4+ T- cell and viral load assessments. Ribonucleic acid (RNA) was extracted from stored plasma and the protease gene amplified, sequenced and analyzed for subtype and drug resistance mutations using the Stanford HIV drug resistance database. RESULTS: Out of the 96 PLWH, 37 experienced virological failure with 8 patients' samples successfully sequenced. The predominant HIV-1 subtype identified was CRF02_AG (6/8, 75.0%) with 12.5% (1/8) each of CFR06_cpx infection and one case unable to subtype. The major PI resistance mutations identified were; M46I, I54V, V82A, I47V, I84V and L90M. CONCLUSIONS: Persons living with HIV who had experienced virologic failure in this study harboured drug resistance mutations to PI, thus compromise the effectiveness of the drugs in the second line. Resistance testing is strongly recommended prior to switching to a new regimen. This will help to inform the choice of drug and to achieve optimum therapeutic outcome among PLWH in Ghana.

Resistant C. albicans implicated in recurrent vulvovaginal candidiasis (RVVC) among women in a tertiary healthcare facility in Kumasi, Ghana.

BACKGROUND: Vulvovaginal candidiasis is a common fungal infection that affects the female lower genital tract. This study determined the major risk factors associated with vulvovaginal infection (VVI) in the Ashanti region of Ghana and also determined the antifungal resistance patterns of Candida albicans isolates to some antifungals. METHODS: Three hundred and fifty (350) high vaginal swab (HVS) samples were collected from women who presented with signs and symptoms of VVI. A structured questionnaire was administered to one hundred and seventy-two (172) of the women. HVS samples were cultured on Sabouraud dextrose agar with 2% chloramphenicol. The polymerase chain reaction was employed to confirm C. albicans. Antifungal susceptibility testing was performed and the susceptibility of C. albicans isolates to fluconazole, clotrimazole, amphotericin B, nystatin, miconazole and 5-flurocytosine were assessed. RESULTS: Vaginal infection was most prevalent amongst females in their reproductive age (21 to 30 years; 63.0%). The study found a significant association between vaginal infections and some risk factors such as sexual practices (p < 0.001), antibiotic misuse (p < 0.05), poor personal hygiene (p < 0.005) and birth control methods (p < 0.049). Out of the 350 HVS samples collected, 112 yielded yeast cells with 65 (58%) identified as C. albicans. The C. albicans isolates were resistant to 5' flucytosine (100%), fluconazole (70%), voriconazole (69.2%), miconazole (58.5%) and nystatin (49.2%). C. albicans isolates were more susceptible to amphotericin B (53.8%) and clotrimazole (45.1%), although an appreciable number of isolates showed resistance (46.1% and 52.3%, respectively). CONCLUSION: There should be nationwide education on all associated risk factors of VVI. Also, use of the various antifungal agents in vaginal candidiasis should proceed after antifungal susceptibility testing to ensure efficacious use of these agents.

Diversity and antibiograms of bacteria isolated from cutaneous leishmaniasis wounds in the Nkwanta South District of Ghana.

Leishmaniasis is a vector-borne disease caused by an intracellular protozoan parasite. The presence of secondary bacterial infections in cutaneous leishmaniasis wounds exacerbate lesion development and could lead to delay in the healing process. This study sought to determine the resistance patterns of bacteria co-infecting cutaneous leishmaniasis wounds from selected communities in the Nkwanta district. Various bacteria were isolated and characterized from exudates obtained from wound swabs collected with sterile cotton tipped applicators. Confirmation of bacterial identity was done using the analytical profile index and the matrix-assisted laser desorption/ionization time of flight mass spectrometry. Antibiotic susceptibility tests were performed using agar disc diffusion method according to the Clinical and Laboratory Standards Institute breakpoint values. A total of eleven (11) secondary bacterial species (spp) were isolated from the 33 wound samples that tested positive for Leishmania kinetoplast DNA, among which Staphylococcus aureus was the most predominant (31%). The pathogenic bacteria that colonized the wounds included Bacillus subtilis (23.8%), Pantoea species (11.9%), Klebsiella pneumoniea (7.1%), Enterobacter cloacae (7.1%), Aeromonas species (4.8%), Serratia marcescens (4.8%), Serratia liquefacien (2.4%), Serratia plymutheca (2.4%), Providencia rettgeri (2.4%) and Cronobacter species (2.4%). Most of the isolates were resistant to beta-lactam antibiotics and the third-generation cephalosporin. Notably, 84.6% of the S. aureus isolates were methicillin and ciprofloxacin resistant whilst 92.3% were resistant to ampicillin. About sixty-nine percent (69.2%) showed intermediate susceptibility to Erythromycin. Additionally, S. plymutheca was resistant to all the test antibiotics. This study suggests colonization of cutaneous leishmaniasis wounds with varied bacterial species that are mostly resistant to beta-lactam group of antibiotics.

Multi-drug resistant bacteria isolates from lymphatic filariasis patients in the Ahanta West District, Ghana.

BACKGROUND: Antimicrobial resistance is associated with increased morbidity in secondary infections and is a global threat owning to the ubiquitous nature of resistance genes in the environment. Recent estimate put the deaths associated with bacterial antimicrobial resistance in 2019 at 4.95 million worldwide. Lymphatic filariasis (LF), a Neglected Tropical Disease (NTD), is associated with the poor living in the tropical regions of the world. LF patients are prone to developing acute dermatolymphangioadenitis (ADLA), a condition that puts them at risk of developing secondary bacterial infections due to skin peeling. ADLA particularly worsens the prognosis of patients leading to usage of antibiotics as a therapeutic intervention. This may result in inappropriate usage of antibiotics due to self-medication and non-compliance; exacerbating antimicrobial resistance in LF patients. In this perspective, we assessed the possibilities of antimicrobial resistance in LF patients. We focused on antibiotic usage, antibiotic resistance in Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa isolates and looked at genes (mecA and Extended-spectrum beta-lactamase [blaCTX-M, blaSHV and blaTEM]) coding for resistance in multi-drug resistant (MDR) bacterial isolates. RESULTS: Of the sixty (60) participants, fifty-four (n = 54, 90%) were within 31-60 years of age, twenty (n = 20, 33.33%) were unemployed and thirty-eight (n = 38, 50.67%) had wounds aged (in months) seven (7) months and above. Amoxicillin (54%) and chloramphenicol (22%) were the most frequently used antibiotics for self-medication. Staphylococcus aureus isolates (n = 26) were mostly resistant to penicillin (n = 23, 88.46%) and least resistant to erythromycin (n = 2, 7.69%). Escherichia coli isolates (n = 5) were resistant to tetracycline (n = 5, 100%) and ampicillin (n = 5, 100%) but were sensitive to meropenem (n = 5, 100%). Pseudomonas aeruginosa isolates (n = 8) were most resistant to meropenem (n = 3, 37.50%) and to a lesser ciprofloxacin (n = 2, 25%), gentamicin (n = 2, 25%) and ceftazidime (n = 2, 25%). Multi-drug resistant methicillin resistant Staphylococcus aureus (MRSA), cephalosporin resistant Escherichia coli. and carbapenem resistant Pseudomonas aeruginosa were four (n = 4, 15.38%), two (n = 2, 40%) and two (n = 2, 25%) respectively. ESBL (blaCTX-M) and mecA genes were implicated in the resistance mechanism of Escherichia coli and MRSA, respectively. CONCLUSION: The findings show presence of MDR isolates from LF patients presenting with chronic wounds; thus, the need to prioritize resistance of MDR bacteria into treatment strategies optimizing morbidity management protocols. This could guide antibiotic selection for treating LF patients presenting with ADLA.

Aspilia africana (Pers.) C.D. Adams and Manihot esculenta Crantz Exhibit Antibacterial Activity against Resistant Salmonella typhi Strains.

Typhoid fever, caused by Salmonella typhi, has plagued underdeveloped countries for many years. Recently, there has been a surge in S. typhi strains identified to be multidrug-resistant in endemic areas. Aspilia africana and Manihot esculenta have been reported to exhibit activity against S. typhi; however, this study aimed to investigate the effect of A. africana and M. esculenta against resistance strains of S. typhi. The leaves of the plants were extracted using distilled water (hot (AQH) and cold (AQC)), methanol (MET), ethyl acetate, and petroleum ether. The extracts were screened in vitro for anti-Salmonella effects against fourteen S. typhi isolates (five multidrug-resistant (MDRST), five ciprofloxacin-resistant (CRST), three nalidixic acid-resistant (NARST), and one sensitive isolate (SS)) using agar well diffusion and microbroth dilution methods. Phytochemical screening showed the presence of tannins, saponins, glycosides, and flavonoids in all polar solvent extracts. Alkaloids were found in all extracts, while triterpenoids were present in all except the aqueous extracts. The AQC of A. africana had the best inhibitory effect on the MDRST and NARST with diameter zones of inhibitions (DZOIs) of 40.0 ± 2.08 mm and 34.0 ± 3.22 mm, respectively. Methanol extract of A. africana had the best inhibitory effect on CRST and SS with DZOIs of 34.0 ± 2.08 and 43.0 ± 3.06 mm, respectively. The AQC and AQH of A. africana and AQH of M. esculenta produced the best MICs and MBCs of 2.5 and 5.0 mg/mL against the MDRST. There was no significant difference in ZOIs of the different solvent extracts against test organisms at p < 0.05. Gas chromatography-mass spectrometry analysis of the extracts showed compounds such as n-hexadecanoic acid, 9,12,15-octadecatrienoic acid (9.55%), and 2H-benzo[F]oxireno[2,3-E]benzofuran-8(9H)-one in the A. Africana extracts and D-mannose, 3-nitrophenyl, methanol acetate (ester), and 9-octadecenamide in the M. esculenta extracts. The leaves of M. esculenta and A. Africana are effective against multidrug-resistant Salmonella isolates.

Gut-Brain-axis: effect of basil oil on the gut microbiota and its contribution to the anticonvulsant properties.

BACKGROUND: Epilepsy is a chronic neurological condition that disrupts the normal functioning of the brain and it is characterized by seizures. Research suggests the involvement of the Gut-Brain axis in epilepsy. This study seeks to determine the role of the gut microbiota in the anticonvulsant effect of basil oil (BO) using antibiotic-depleted and altered germ-free mice against naïve mice in Pentylenetetrazole (PTZ) induced seizure model. There is an ever growing interest in improvement of treatment outcomes in epilepsy and also in the development of newer therapeutic options, especially in the population of patients that do not attain seizure relief from available antiseizure medications (ASMs). According to research, gut microbiota can alter brain function and development. Increasing evidence suggests disrupting the delicate symbiotic balance existing between the gut and brain results in disease conditions. Also, the oil from Ocimum basilicum L., (BO) has been proven scientifically to significantly block clonic seizures induced by PTZ and picrotoxin in seizure models. METHODS: The microbiota of mice were depleted or altered by administering cocktail antibiotics and individual antibiotics respectively. DNA was isolated from mouse stool, and then the 16S ribosomal ribonucleic acid (16S rRNA) gene was quantitatively amplified using reverse transcription-polymerase chain reaction (RT-PCR). Amplicons were sequenced to determine the phylogenetic make-up of the bacteria involved. Metabolic profiles of the serum and stool of mice were determined using Proton (1H) Nuclear Magnetic Resonance (NMR) spectroscopy. RESULTS: Cocktail antibiotic pre-treatment significantly reversed the anticonvulsant effect of BO by increasing frequency and duration of seizures but did not affect latency to seizure. In mice pre-treated with single antibiotics, the anticonvulsant effect of BO was lost as latency to seizures, frequency and duration of seizures increased compared to mice that received only BO. Assessment of the phylogenetic make-up of the microbiota in antibiotic pre-treated mice showed a distorted composition of the microbiota compared to the control group. CONCLUSION: Depletion of the microbiota significantly reversed the anticonvulsant actions of BO. The concentrations of short chain fatty acids (SCFAs) was higher in stool than in the serum of the mice. Administration of BO probably does not influence the microbial composition within the mouse microbiota. The elevated ratio of Firmicutes to Bacteroidetes in microbiota-depleted groups might have contributed to the reversal of anticonvulsant actions of BO.

Antimicrobial and Anti-Inflammatory Activities of Pterygota macrocarpa and Cola gigantea (Sterculiaceae).

Pterygota macrocarpa and Cola gigantea are African medicinal plants used in traditional medicine for the treatment of sores, skin infections, and other inflammatory conditions including pains. This study therefore aims at investigating the antimicrobial properties of ethanol leaf and stem bark extracts of P. macrocarpa and C. gigantea using the agar diffusion and the micro-dilution techniques and also determining the anti-inflammatory properties of the extracts of these plants in carrageenan-induced foot edema in seven-day old chicks. The minimum inhibitory concentration of both ethanol leaf and bark extracts of P. macrocarpa against the test organisms was from 0.125 to 2.55 mg/mL and that of C. gigantea extracts was 0.125 to 2.75 mg/mL. Extracts with concentration of 50 mg/mL were most active against the test organisms according to the agar diffusion method. All the extracts of P. macrocarpa and C. gigantea at 30, 100, and 300 mg/kg body weight except ethanol leaf extract of C. gigantea exhibited significant anti-inflammatory effects (P ≤ 0.001).

Sensitivity Patterns, Plasmid Profiles and Clonal Relatedness of Multi-Drug Resistant Pseudomonas aeruginosa Isolated From the Ashanti Region, Ghana.

Pseudomonas aeruginosa is a major cause of most opportunistic nosocomial infections in Ghana. The study sought to characterize P. aeruginosa isolates from market environments, poultry farms and clinical samples of patients from 2 district hospitals in the Ashanti region of Ghana. The genetic relatedness, plasmid profiles and antimicrobial sensitivity of the isolates were investigated. Culture based isolation and oprL gene amplification were used to confirm the identity of the isolates. Susceptibility testing was conducted using the Kirby Bauer disk diffusion method. Random whole genome typing of the P. aeruginosa strains was done using Enterobacterial repetitive-intergenic consensus based (ERIC) PCR assay. The most active agents against P. aeruginosa isolates were ceftazidime (90%), piperacillin (85%), meropenem, cefipeme and ticarcillin/clavulanic acid (81.6%). The isolates were most resistant to gentamycin (69%), ciprofloxacin (62.1%), ticarcillin (56.3%) and aztreonam (25%). About 65% (n = 38) of the multi-drug resistant (MDR) P. aeruginosa isolates harbored 1 to 5 plasmids with sizes ranging from 2 to 116.8 kb. A total of 27 clonal patterns were identified. Two major clones were observed with a clone showing resistance to all the test antipseudomonal agents. There is therefore a need for continued intensive surveillance to control the spread and development of resistant strains.

Evaluation of Dermal Toxicity and Wound Healing Activity of Cnestis ferruginea Vahl ex DC.

Cnestisferruginea is a tropical plant, which is traditionally used in the treatment and management of various conditions including skin infections and wounds. The aim of this study was to investigate the dermal toxicity and wound healing potential of C. ferruginea. Ten millimeter full-thickness mucosal wounds were created on the dorsal midportion of the Sprague Dawley rats. Wounds were treated with 10, 5, and 2.5% w/w aqueous creams, prepared from the methanol extract of the root bark of C. ferruginea (CFM). The wound tissues were harvested on day 21 for histology studies. Compared with the untreated group, 10, 5, and 2.5% w/w CFM-treated wounds significantly reduced the wound size over the study period (P < 0.0001). Tissue histology revealed a healed wound with well-regenerated collagen and skin appendages with no pus cells. A skin irritation test was conducted on CFM, as well as the dermal toxicity of CFM was determined in the repeated dose and acute dermal toxicity bioassays. These tests revealed that CFM showed no toxic effect on the skin and showed that CFM was not a skin irritant. C. ferruginea exhibited wound healing activity, which gives credence to its folkloric use.

Prevalence and Phenotypic and Genotypic Resistance Mechanisms of Multidrug-Resistant Pseudomonas aeruginosa Strains Isolated from Clinical, Environmental, and Poultry Litter Samples from the Ashanti Region of Ghana.

Background: Antibiotic resistance in bacteria is a major global health challenge. Reports on the prevalence of multidrug-resistant P. aeruginosa, a common pathogenic bacterium implicated in nosocomial infections and poultry diseases, are limited in Ghana. This study therefore sought to determine the prevalence of P. aeruginosa from hospitals, poultry farms, and environmental samples from the Ashanti region of Ghana. Methodology. Stool, urine, and blood samples from 364 patients from two hospitals in the Ashanti region of Ghana were randomly sampled. P. aeruginosa was isolated and confirmed using routine selective media and PCR-based oprL gene amplification. The Kirby-Bauer disk diffusion method employing EUCAST breakpoint values was used to identify multidrug-resistant strains. The occurrence of common antibiotic inactivating enzymes and resistance encoding genes and the assessment of strain efflux capacity were investigated with double disc synergy test (DDST), imipenem-EDTA synergy test, phenylboronic acid test, D-test, routine PCR, and ethidium bromide agar-cartwheel method. Results: A total of 87 (9.7%, n = 87/900) P. aeruginosa isolates were confirmed from the samples. 75% (n = 65/87) were resistant to more than one group of antipseudomonal agents, while 43.6% (n = 38/87) were multidrug-resistant (MDR). High prevalence of extended spectrum ß-lactamases (84.2%), metallo-ß-lactamases (34.1%), and AmpC inducible cephalosporinases (50%) was observed in the MDR strains. About 57.8% of the MDR strains showed moderate to very high efflux capacity. Class 1 integrons were detected in 89.4% of the MDR isolates but ß-lactamase encoding genes (bla SHV , bla TEM , bla CTX-M , bla VIM , and bla IMP ) were not detected. Conclusion: Surveillance of antibiotic-resistant strains of bacteria should be routinely conducted in clinical and veterinary practice in Ghana to inform selection of antibiotics for therapeutic use.

In Vitro Anthelmintic Activity of Ethanol Stem Bark Extract of Albizia ferruginea (Guill. & Perr.) Benth.

Albizia ferruginea (Guill. & Perr.) Benth bark is used in the traditional medicine as a vermifuge. This study sought to determine the anthelmintic activity of the stem bark extract of Albizia ferruginea. The powdered A. ferruginea stem bark was extracted with ethanol. Phytochemical screening was carried out on A. ferruginea ethanol extract (AFE) and then screened for its anthelmintic property against Pheretima posthuma and Haemonchus contortus using the adult motility assay. The effect of AFE and its fractions on the anthelminthic activity of mebendazole and albendazole were also determined using the adult worm (P. posthuma) motility assay. AFE showed a dose-dependent anthelmintic activity against P. posthuma and H. contortus. The least concentration of AFE (0.5 mg/mL) paralyzed and killed P. posthuma within 272.50 ± 12.42 min and 354.50 ± 5.06 min of exposure, respectively. AFE at the least test concentration (0.14 mg/mL) caused paralysis and induced death of H. contortus, after at 63.50 ± 2.98 and 254.96 ± 2.44 min of exposure, respectively. AFE extract at 0.25 and 0.125 mg/mL increased the paralytic and helminthicidal activities of albendazole. The paralytic and helminthicidal activities of mebendazole were reduced when combined with AFE (0.25 and 0.125 mg/mL). Among the three fractions obtained from AFE, the methanol fraction showed the highest anthelmintic activity. The methanol fraction at 0.5 mg/mL caused paralysis after 69.90 ± 0.15 min and death of worm after 92.53 ± 0.74 min of exposure. The petroleum ether and ethyl acetate fractions showed relatively low anthelmintic activity. Phytochemical screening of AFE revealed the presence of tannins, saponins, glycosides, alkaloids, and coumarins. The results from this study show that A. ferruginea possesses anthelmintic activity which gives credence to its folkloric use.

Prevalence and antibiotic resistance of coagulase-negative Staphylococci isolated from poultry farms in three regions of Ghana.

The use of antibiotics in animal production has been associated with the development and spread of antibiotic-resistant organisms including commensals. Coagulase-negative Staphylococcus (CoNS) species, which were until recently considered non-pathogenic, have been associated with opportunistic infections and high resistance to several antibiotics. This study sought to determine the prevalence, identity, and phenotypic resistance of coagulase-negative Staphylococcus spp. isolated from some selected poultry farms and farm workers in the Ashanti, Brong Ahafo, and Greater Accra regions of Ghana. Poultry litter samples and oral swabs of poultry farm workers were collected, from which bacterial species were isolated, identified, and analyzed. Various selective media were used for the presumptive identification of the different species. Confirmation of bacterial identity was done using matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry. Antibiotic susceptibility testing of the isolates was performed using the Kirby-Bauer disk diffusion method. Zones of growth inhibition were interpreted based on the European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines. Two hundred and fifty-six coagulase-negative Staphylococcus spp., comprising S. sciuri (42.97%), S. lentus (35.94%), S. gallinarum (6.64%), S. xylosus (4.30%), S. haemolyticus (3.91%), S. saprophyticus (1.95%), and S. cohnii (0.39%) were confirmed by MALDI-TOF. CoNS were isolated from samples from the Brong Ahafo (48.83%), Ashanti (33.59%), and Greater Accra (17.78%) regions. Isolates from poultry litter constituted 55.47%, and farm workers 44.53%. All the isolates were susceptible to amoxicillin/clavulanic acid and amikacin. The isolates exhibited high resistance toward tetracycline (57.03%), doxycycline (43.75%), and oxacillin (43.36%). Multi-drug resistance (MDR) was observed in 19.14% of the isolates. MDR was higher in isolates obtained from poultry farm workers (61.22%) than isolates from poultry litter (38.78%). The above findings call for stricter monitoring of antibiotic usage in both animal production and in humans.

Antibiotic Resistance Patterns of Escherichia coli Isolates from Hospitals in Kumasi, Ghana.

Nosocomial infections are infections acquired by a patient as a result of treatment in a hospital or healthcare service providing center and symptoms occurs within a short period of hospitalization. The study was to determine the antibiotic resistance patterns of Escherichia coli isolated from Kumasi-South, Tafo and Suntreso Hospitals, Kumasi, Ghana. Total of 600 swabs samples from the hospitals were collected between January and June, 2010. The isolates were identified using morphological and biochemical means. A total of 97 E. coli isolates were obtained from the hospitals. Beds in hospital wards had the highest number of E. coli strains (53.6%), followed by floors (20.6%) while drainages had the least isolates (3.1%). Majority of the E. coli isolates (90.7%) exhibited resistance to ampicillin while 6.2 and 3.1% showed intermediate and sensitive respectively. Co-trimoxazole, 78.4% of the isolates were resistant while 9.3 and 12.4% exhibited intermediate and sensitive responses respectively. E. coli isolates (28.6 to 46.4%) were resistant to gentamicin, ciprofloxacin and ceftriaxone while 14.4 to 47.4% gave intermediate responses. Most isolates (80.4%) exhibited multi-drug resistance. There is a need to observe proper personal hygiene, use of effective disinfectants and proper disposal of contaminated/pathogenic materials in these hospitals to control nosocomial infections.