Direct and Residual Microbicidal Efficacy of Various Antiseptics against Multi-Drug Resistant Bacteria

Background: Infections in ICU’s patients are known to often originate from the colonization of wounds by the patient’s endogenous microbiota, and to eventually lead to secondary sepsis. Aim: to compare in vitro the direct and residual effects after different exposure times of 4% chlorhexidine, and of 0.1% and 0.04% polyhexanide (in gel and solution forms), on ATCC-microorganisms, and too, on bacterial strains obtained from ICU patients. Methods: We used wild multi-drug resistant strains recently obtained from the wounds of patients hospitalized at ICU and reference strains from the American Type Culture Collection (ATCC). Chlorhexidine 4% was studied as a reference solution. The direct and residual effects of the 0.1% and 0.04% polyhexanide, in gel and solution forms, were analyzed using cotton germ carriers. To evaluate the direct effect, we exposed the strains to the antiseptic. To assess the residual effect, the germ-carriers were impregnated with antiseptic and were allowed to dry before we contaminated them. We inoculated the germ carriers in a culture medium with an inhibitor of antiseptic effect to count the number of surviving microorganisms. Findings: 0.1% Polyhexanide solution proved a direct and residual efficacy after 24 hours equivalent to 4% chlorhexidine. Is very important to highlight that this great efficacy did not change according to whether they were ATCC or multidrug-resistant strains. Conclusions: 0.1% polyhexanide demonstrated a great direct and residual efficacy (like 4% chlorhexidine), against multi-drug resistant strains isolated from ICU’s patients. Moreover, due to its few cytotoxicity against keratinocytes and fibroblasts can be an optimal antiseptic for burns, wounds or ulcers.

Analysis of Multi-Drug Resistant Organism Surveillance and Antimicrobial Resistance Early Warning in a Hospital in 2022

Objective:To determine the clinical distribution of multi-drug resistant organism(MDRO)in Jiangyan Hospital and the monitoring and warning of drug-resistance bacteria to provide an important basis for guiding the application of broad-spectrum antibiotics in clinical treatment and reducing the occurrence of nosocomial infection.Methods:Retrospective screening and analysis were conducted on the pathogenic strains of hospitalized patients in our hospital in 2022.Results:A total of 2,769 strains of pathogenic bacteria and 390 strains of MDRO were detected and isolated in our hospital in 2022;the detection rate of MDRO was 14.08%.A total of 516 strains(18.64%)Klebsiella pneumoniae(KP)and 62 strains(12.02%)of carbapenem-resistant Klebsiella pneumoniae(CR-KP)were detected;436 strains(15.75%)of Escherichia coli(ECO)were detected,including 8 strains(1.83%)of CR-ECO;342 strains(12.35%)of Pseudomonas aeruginosa(PA)and 116 strains(33.92%)of CR-PA were detected;there were 194 strains(7.01%)of Acinetobacter baumannii(AB),among which 125 strains(64.43%)were CR-AB;there were 291 strains(10.51%)of Staphylococcus aureus,among which 79 strains(27.15%)of methicillin-resistant Staphylococcus aureus(MRSA)were detected;78 strains(2.82%)of Enterococcus faecalis were detected,and vancomycin-resistant enterococcus(VRE)was not detected.The first five MDROs were CR-AB,CR-PA,MRSA,CR-KP,and CR-ECO.The top five departments with the highest MDRO detection rate in 2022 were the ICU(37.44%),the Pulmonology Department(ward 13;31.03%),the Department of Rehabilitation(ward 5;6.67%),the Department of Neurosurgery(ward 11;4.62%),and the Department of General Surgery(ward 10;3.59 The resistance rate of antibacterial drugs is divided into four levels for early warning:30%to 40%,41%to 50%,51%to 75%,and 75%or more.Conclusion:Our hospital should strengthen the monitoring of antimicrobial resistance warning related to MDRO and the abuse of antimicrobial drugs.Based on the results of drug sensitivity and antimicrobial resistance warning,the use of antibiotics should be standardized in clinical practice to reduce nosocomial infection。

Detection of Multi-drug Resistant Acinetobacter Lwoffii Isolated from Soil of Mink Farm

There were 4 Acinetobacter lwoffii obtained from soil samples.The antimicrobial susceptibility of the strains to 16 antimicrobial agents was investigated using K-B method.Three isolates showed the multi-drug resistance.The presence of resistance genes and integrons was determined using PCR.The aadA 1,aac（3＇）-IIc,aph（3＇）-VII,aac（6＇）-Ib,sul2,cat2,floR,and tet（K）genes were detected,respectively.

Analysis of Clinical Symptoms Improvement in Treatment of Severe Pneumonia Caused by Multi-drug Resistant Bacterial Infection by Bronchoscopy Alveolar Lavage

Objective:To explore the effects of bronchoscopy alveolar lavage in the treatment of severe pneumonia caused by multiple drug bacterial infection.Methods:A total of 84 patients with severe pneumonia infected by multi-drug resistant bacteria were randomly selected from Kunming Second People's Hospital,which is our hospital from January 2019 to December 2020 for this research.They were divided into a reference group and a study group using a digital table method,with 42 cases in each group.The reference group was given the routine treatment,the research group performed bronchoscopy alveolar lavage on this basis to observe the curative effect.Results:Before treatment,there was no significant difference in serum factor indexes and blood gas analysis indexes between the two groups of patients,P>0.05.After treatment,the time of fever,cough,moist rales disappearing and infection control time in the study group were shorter than those in the reference group,P<0.05.Serum tumor necrosis factorα(TNF-α),interleukin-6(IL-6)and interleukin-8(IL-8)levels were lower than the reference group,and the arterial partial pressure of oxygen(PO2)and oxygen saturation(SO2)were both lower than the reference group,P<0.05.The effective rate of the treatment in the study group was higher than that in the reference group,P<0.05.Conclusion:Bronchoscopic alveolar lavage treatment can effectively improve the clinical symptoms of patients with severe pneumonia caused by multi-drug resistant bacterial infection,and the effects are significant.

Acceleration of apoptosis by transfection of bak gene in multi-drug resistant (MDR) bladder cancer cellsI

Objective To observe the effects of bak gene on killing MDR bladder cancer cells and to study its mechanisms. Methods Bak gene was transfected into MDR bladder cancer cells by liposome. The mRNA of bak and bcl-2 were detected by in situ hybridization. The protein of bak and bcl-2 were detected by SABC immunohistochemistry. The growth rate of human bladder cancer cells was studied by constructing the growth curve, cell apoptosis being observed by flow cytometry, and the outline of cells observed by fluorescence stain. Results The expression of bak mRNA was positive in EJ/bak cells (64% ,P【0.05).Bak protein expression of EJ/bak cells was positive (60 % ) and bcl-2 protein expression was de creased (P【0.05). The growth of MDR bladder cancer cells was significantly inhibited by 32% after bak gene was transfected (P 【 0. 05 ). Apoptosis cells increased significantly. The apoptosis rate was 35 %. Apoptotic bodies can be found in these cells on fluorescence stain. Conclusion Bak gene could inhibit the growth

Antimicrobial activity of green tea extract against isolates of methicillin-resistant Staphylococcus aureus and multi-drug resistant Pseudomonas aeruginosa

Objective:To evaluate antibacterial activity of the Indonesian water soluble green tea extract,Camellia sinensis,against clinical isolates of methicillin-resistantStaphylococcus aureus (S.aureus)(MRSA)and multi-drug resistant Pseudomonas aeruginosa(MDR-P.aeruginosa).Methods:Antimicrobial activity of green tea extract was determined by the disc diffusion method and the minimum inhibitory concentration(MIC)was determined by the twofold serial broth dilutions method.The tested bacteria using in this study were the standard strains and multi-drug resistant clinical isolates of S.aureus and P.aeruginosa,obtained from Laboratory of Clinical Microbiology,Faculty of Medicine,University of Indonesia.Results:The results showed that the inhibition zone diameter of green tea extracts forS.aureus ATCC 25923 and MRSA were(18.970依0.287)mm,and(19.130依0.250)mm respectively.While the inhibition zone diameter forP.aeruginosa ATCC 27853 and MDR-P.aeruginosawere(17.550依0.393)mm and(17.670依0.398)mm respectively.The MIC of green tea extracts againstS.aureus ATCC 25923 and MRSA were 400μg/mL and 400μg/mL,respectively,whereas the MIC for P.aeruginosa ATCC 27853 and MDR-P.aeruginosawere 800μg/mL,and 800μg/mL,respectively.Conclusions:Camellia sinensisleaves extract could be useful in combating emerging drug-resistance caused by MRSA andP.aeruginosa.

Cytotoxic and antibacterial substances against multi-drug resistant pathogens from marine sponge symbiont:Citrinin,a secondary metabolite of Penicillium sp.

Objective:To Isolate,purify,characterize,and evaluate the bioaclive compounds from the sponge-derived fungus Penicillium sp.FF001 and to elucidate its structure.Methods:The fungal strain FF001 with an interesting bioactivity profile was isolated from a marine Fijian sponge Melophlus sp.Based on conidiophores aggregation,conidia development and mycelia morphological characteristics,the isolate FF001 was classically identified as a Penicillium sp.The bioactive compound was identified using various spectral analysis of UV,high resolution electrospray ionization mass spectra,1H and 13C NMR spectral data.Further minimum inhibitory concentrations(MICs)assay and brine shrimp cytotoxicity assay were also carried out to evaluate the biological properties of the purified compound.Results:Bioassay guided fractionation of the EtOAc extract of a static culture of this Penicillium sp.by different chromatographic methods led the isolation of an antibacterial,anticryptococcal and cytotoxic active compound,which was identified as citrinin(1).Further,citrinin(1)is reported for its potent antibacterial activity against methicillin-resistant Staphylococcus aureus(S.aureus),rifampicin-resistant 5.aureus,wild type S.aureus and vancomycin-resistant Enterococcus faecium showed MICs of 3.90,0.97,1.95 and7.81μg/mL,respectively.Further citrinin(1)displayed significant activity against the pathogenic yeast Cryptococcus neoformans(MIC 3.90μg/mL),and exhibited cytotoxicity against brine shrimp larvae LD_(50)of 96μg/mL.Conclusions:Citrinin(1)is reported from sponge associated Penicillium sp.from this study and for its strong antibacterial activity against multi-drug resistant human pathogens including cytotoxicity against brine shrimp larvae,which indicated that sponge associated Penicillium spp.are promising sources of natural bioactive metabolites.

<i>In Vitro</i>Antibacterial Activity of Flavonoid Extracts of Two Selected Libyan Algae against Multi-Drug Resistant Bacteria Isolated from Food Products

This study aimed to evaluate the antibacterial activity of flavonoids extracted from two Libyan brown algae namely Cystoseira compressa and Padina pavonica using microwave-assisted extraction method against pathogenic bacteria isolated from meat, meat products, milk and dairy products (Staphylococcus aureus subsp. aureus (5 isolates), Bacillus cereus (3 isolates), Bacillus pumilus (1 isolate), Salmonella enterica subsp. enteric (4 isolates) and Enterohaemor-rhagic Escherichia coli O157 (EHEC O157) (4 isolates)). All of these isolates were muti-drug resistant with high MAR index. The results showed that C. compressa extract exhibited better and stronger antibacterial activities against the seventeen tested isolates with inhibition zones diameter ranged from 14 - 22 mm compared to P. pavonica extract which showed positive effect against 9 isolates with low inhibition zone ranged from 11 - 16.5 mm. Flavonoids extracted from C. compressa also displayed the best spectrum of bactericidal effect with a ratio MBC/MIC ≤ 4 obtained on all susceptible tested bacterial strains. Flavonoids and proanthocyanidins significantly contributed to the antibacterial properties. The mode of action of these active extracts is under investigation.

Drug Resistance Pattern in Pulmonary Tuberculosis Patients and Risk Factors Associated with Multi-Drug Resistant Tuberculosis

Introduction: Anti-tuberculosis drug resistance is a major problem in tuberculosis (TB) control programme, particularly multi-drug resistance TB (MDR-TB) in Nepal. Drug resistance is difficult to treat due to its associated cost and side effects. The objective of this study was to assess the drug resistance pattern and assess risk factor associated with MDR-TB among pulmonary tuberculosis patients attending National Tuberculosis Center. Methodology: The comparative cross sectional study was conducted at National Tuberculosis Center during August 2015 to February 2015. Early morning sputum samples were collected from pulmonary tuberculosis suspected patients and subjected to Ziehl-Neelsen staining and fluorochrome staining and culture on Lowenstein-Jensen (LJ) medium. Drug Susceptibility test was performed on culture positive isolates by using proportion method. Univariate and multivariate analysis was computed to assess the risk factors of MDR-TB. Results: Out of 223 sputum samples, 105 were fluorochrome staining positive, 85 were ZN staining positive and 102 were culture positive. Out of 102 culture positive isolates, 37.2% were resistance to any four anti-TB drugs. 11 (28.9%) were initial drug resistance and 28 (43.7%) were acquired drug resistance. The overall prevalence of MDR-TB was 11.7%, of which 2 (5.3%) were initial MDR-TB and 10 (15.6%) were acquired MDR-TB. Univariate and multivariate analysis showed female were significantly associated (P = 0.05) with MDR-TB. Conclusion: Drug resistance TB particularly MDR-TB is high. The most common resistance pattern observed in this study was resistance to both isoniazid and rifampicin. Female were found to be associated with MDR-TB. Thus, early diagnosis of TB and provision of culture and DST are crucial in order to combat the threat of DR-TB.

Predominance of Multi-Drug Resistant <i>Klebsiella pneumonia</i>and Other Gram Negative Bacteria in Neonatal Sepsis in Equatorial Guinea

The study was conducted on new-born babies in whom septicemia was suspected, to determine the prevalence of bacterial strains isolated and their sensitivity to antimicrobial drugs. The study was carried out at La Paz Medical Center, Microbiology section, Malabo, Equatorial Guinea from August 2013 to October 2015. Out of 293 septicemia suspected cases, 29 (10%) blood cultures were positive, 28 with bacterial growth and 1 with growth of Candida sp. The mortality rate of neonates caused by Gram negative bacterial sepsis was 34.7%. Among the Gram negative bacteria (24 isolates), the most common types were Klebsiella pneumoniae (16 = 69.6%), followed by Escherichia coli (4 = 17.4%) and Acinetobacter species (4 = 17.4%). Four Gram positive bacteria were also isolated and identified all ascoagulase-negative staphylococci. All the Klebsiella pneumoniae isolates and Acinetobacter species demonstrated Multi Drug Resistance against different antibiotics with Extended-spectrum β-lactamase (ESBL) activity. The most frequent causative agent of bacterial sepsis in new-born children was Klebsiella pneumoniae. An alarming level of Multi Drug Resistance (MDR) Klebsiella pneumoniae strains to the first choice antibiotic treatment was observed.

Natural Remedies against Multi-Drug Resistant <i>Mycobacterium tuberculosis</i>

Tuberculosis (TB), caused by Mycobacterium tuberculosis is an infectious deadly disease and the treatment of which is one of the most severe challenges at the global level. Currently more than 20 chemical medications are described for the treatment of TB. Regardless of availability of several drugs to treat TB, the causative agent, M. tuberculosis is nowadays getting resistant toward the conventional drugs and leading to conditions known as Multidrug-resistant tuberculosis (MDR-TB) and extensively drug resistant tuberculosis (XDR-TB). This situation has terrified the global health community and raised a demand for new anti-tuberculosis drugs. Medicinal plants have been used to cure different common as well as lethal diseases by ancient civilizations due to its virtue of variety of chemical compounds which may have some important remedial properties. The aim of the present review is to focus the anti-tubercular medicinal plants native to India as well as the plants effective against MDR or XDR-TB across the globe. In the present review, we have addressed 25 medicinal plants for TB and 16 plants effective against MDR-TB testified from India and 23 herbal plants described for MDR-TB across the world during 2011-2015. These herbal plants can serve as promising candidates for developing novel medications to combat multidrug resistant M. tuberculosis.

Inhibitory Activities of Lactic Acid Bacteria against Multi-Drug Resistant Uropathogenic <i>Staphylococcus saprophyticus</i>Isolated from Symptomatic Women in Lagos, Nigeria

The uropathogenic Staphylococcus saprophyticus is reported severally to be resistant to the drugs often used empirically for treatment of urinary tract infections (UTIs). Their ability to exhibit resistance to multiple drugs is a great deal of threat to successes recorded in the management of UTIs caused by this pathogen. Lactic acid bacteria (LAB) have been demonstrated to exhibit antimicrobial activities but studies about their prospect against multi-drug resistant S. saprophyticus are quite few. This study therefore investigated activities of LAB against the multi-drug resistant S. saprophyticus recovered from urine samples of symptomatic women. The three different species of LAB (Lactobacillus fermentum BTA 62, Lactobacillus johnsonii BTA 86 and Weissella confusa BTA 40) previously isolated and identified by 16S rRNA sequencing were selected based on their history of antimicrobial activities. Their metabolites were employed in the antagonistic assays against six (6) multi-drug resistant test pathogens recovered from urine samples of symptomatic, non-pregnant women attending clinics in Lagos, Nigeria and the control (S. saprophyticus subs bovis strain DSM 18669) following standard procedures. The pathogens showed resistances to almost all the antibiotics except levofloxacin, ciprofloxacin, imipenem while the control showed resistance to three. The LAB, L. fermentum inhibited five (83.3%) of the pathogens with zone diameter of 12 - 17 mm, followed by W. confusa inhibiting three (50%) with 15 - 17 mm. Lactobacillus johnsonii on the other hand, inhibited a pathogen and the control with zones of 13 mm and 14 mm respectively. In conclusion, the extracted metabolites of LAB inhibited the growth of multi-drug resistant clinical isolates of uropathogenic S. saprophyticus and may therefore be potent alternatives to antibiotics.

Isolation and Identification of Multi-Drug Resistant Strains of Non-Lactose Fermenting Bacteria from Clinical Refuses in Major Hospitals of Khyber Pakhtunkhwa, Pakistan

Purpose: The present studied was performed in order to investigate the drug resistance of different non-lactose fermenting gram negative bacteria from clinical isolates. The bacteria were morphologically characterized through culturing and gram staining techniques were used for the identification of different bacterial strains. Methods: A total of 324 samples were collected from patients, after they were diagnosed by physicians at different hospitals at district Peshawar. Samples were morphologically identified by blood agar, MacConkey agar and Eosine Methylene Blue, identified by gram staining techniques. Modified Kirby-Bauer Disc diffusion method was used to test the in-vitro susceptibility of the identified isolates to different antibiotics. Results: The non-lactose fermenting gram negative bacteria were isolated from samples of blood (33.30%), pus/ wound (33.30%), urine (23.30%) and from ascetic/pleural fluids (10.20%). The study revealed that Pseudomonas aeroginosa showed high resistance against Gentamicin (74%) and Aztreonam (74%), followed by Ciprofloxacin (59.20%) and Amikacin (33.30). Tazocin was active as low resistance (18.50%) is shown. More resistance was seen in Morganella morganii against Aztreonam (77.7%) followed by Gentamicin (62.90%), Ciprofloxacin (40.70%). Tazocin show low resistance (3.70%). Multidrug resistant Proteus mirabillis was highly resistance to Gentamicin (66.60%), followed by Aztreonam (62.90%), Amikacin (55.50%), Ciprofloxacin (40.20%) and low resistance to Tazocin was (22.20%). Salmonella typhi demonstrated high resistance against Amikacin (62.90%), followed by Aztreonam (48.10%), Tazocin (40.70%). Gentamicin showed low resistance (29.60%), and hence it is more active against S. typhi. Conclusions: It can be concluded from the present study that different species of non-lactose fermenting gram negative bacteria have shown a different resistivity pattern. This study is a gate way for better and suitable management strategy for the infections caused by non-Lactose fermenting bacteria in the sampling region.

Occurrence of Multi-Drug Resistant <i>Listeria species</i>in Faecal Samples of Poultry Chickens in Rural Farms in Lagos State, Nigeria

Background: Listeriosis is a common zoonotic disease caused by a foodborne pathogen, Listeria monocytogenes. Poultry meat and products have been established as vehicles of transmission of pathogenic Listeria strains to humans. This study evaluates the occurrence of Listeria species in faeces of poultry chicken in Lagos. Methods: One hundred and fourteen pooled fresh faecal samples from cage-reared broiler chickens were collected from 12 farms in three rural areas in Lagos State from May to August 2019. All samples were analysed for Listeria species detection according to ISO11290-1 standard and confirmed using PCR assay. Susceptibility testing was performed using the Kirby-Bauer disc diffusion technique. Results: Twenty-eight (24.6%) Listeria species were detected from 114 faecal samples. The isolated Listeria species were L. monocytogenes 8 (7.0%), L. ivanovii 9 (7.9%), L. grayi 7 (6.1%) and L. innocua 4 (3.5%). There was no significant difference in the frequency of occurrence of Listeria species across the different locations (X2 = 4.98, p = 0.08). The listeria species were susceptible to Augmentin (96.4%), vancomycin (85.7%) and co-trimoxazole (82.1%), but resistant to ceftazidime (100%), tetracycline (75.0%) and ciprofloxacin (71.4%). Conclusion: This study reveals high occurrence of multi-drug resistant Listeria species in faecal samples of poultry chickens in Lagos state which may be an important vector in the contamination of the environment and transmission of antibiotic resistant Listeria species to consumers.

Bi-Directional Regulation by Chinese Herbal Formulae to Host and Parasite for Multi-Drug Resistant <i>Staphylococcus aureus</i>in Humans and Rodents

A decline in the immunopotential of the host plays acritical factor(s) in the occurrence of infections with methicillin-resistant Staphylococcus aureus (MRSA) or microorganisms by opportunistic infection. In such an infection, no way out for therapeutic concept, therefore bi-directional trial was the final choice. So we selected aformula, Dang Gui Liu Huang Tang (dLHT), which could both augmentimmune factorsin host and exert bacteriostatic effect. We sought to break through the epidemic by MRSA especially in elderly patient, by the fundamental and clinical trial by employing minor TCM, characterizing bidirectional ability of the decoction by western methods. Animal Experiment: Mitomycin-C (MMC)-treated mice with or without the infection of MRSA were made. The experimental design was made up to examine the bacteriostatic action as well as the immunopo-tentiating bias of the promising Chinese herbal medicine, dLHT, which was first proved for its immune potentiating activities as well as their sensitivity to antibiotics, but not direct aseptic effect was clear for MRSA. Both basic and clinical data showed that this formula was effective on repelling from the infectious focus after the treatment of MRSA infection. After the administration of dLHT, the number of white blood cells in MMC-treated mice recovered to 80% of the normal level. In addition, the phagocytic activity of macrophages increased to 70% in the dLHT-treated group, while that of the non-treated group was only 20%. The bactericidal activity also recovered to the level close to the normal value by dLHT. The ratio of neutrophils in the dLHT-administered group increased to 2.2% (normal mice, 2.6%), whereas that in the non-terated group was only 0.5%. The bacterial count in the liver of MRSA-challenged mice reached the peak at six hours after the challenge in both dLHT-treated and non-treated mice. However, the number of bacteria in dLHT group was much greater than that in the non-treated group. The bacterial count in the blood showed an increase 12 and 24 hours after the challenge. Even 24 hours after the challenge, a significant number of bacteria existed in the blood of dLHT-administered group, whereas only a small number of bacteria detectable 6 hours after the challenge and the number gradually decreased thereafter in the dLHT-administered group. MRSA-challenged MMC-treated mice were treated by dLHT, vancomycin, or dLHT and vancomycin. All of non-treated mice died 8 days after the MRSA challenge, whereas the survival rates were 60% after dLHT treatment, 40% after vancomycin treatment, and 80% after dLHT and vancomycin treatment. All of MMC-treated mice, to which the phagocytic cells prepared from MMC-treated mice with dLHT administration had adoptively been transferred, survived from MRSA challenge. On the other hand, the survival rate of MMC-treated mice, to which the lymphocytes prepared from the same mice had adoptively been transferred, was 40%. Clinical Trial: All cases with dLHT treatment showed negative culture results for MRSA after the dLHT administration. The culture generally became negative less than 50 days after the initial administration, whereas one control case needed more than 100 days and the other was dead of the infection. One representative case, who was a 78-year-old woman suffering from hypertension, atrial fibrillation, and cerebral bleeding in the right occipital lobe, infected with MRSA during the antibiotic therapy for Streptococcus pneumoniae. The antibiotic therapy was halted and the dLHT administration started. Three weeks later, the culture result became negative. In addition, serum protein and albumin values also returned to the level that they had had before the infection of MRSA.

Production and purification of a bioactive substance against multi-drug resistant human pathogens from the marine-sponge-derived Salinispora sp.

Objective:To isolate,purify,characterize,elucidate structure and evaluate bioactive compounds from the sponge-derived Salinispora sp.FS-0034.Methods:The symbiotic actinomycete strain FS-0034 with an interesting bioactivity profile was isolated from the Fijian marine sponge Theonella sp.Based on colony morphology and obligatory requirement of seawater for growth,and mycelia morphological characteristics the isolate FS-0034 was identified as a Salinispora sp.The bioactive compound was identified by using various spectral analysis of ultraviolet,high resolution electrospray ionization mass spectroscopy,H nuclear magnetic resonance,correlated spectroscopy and heteronuclear multiple bond coherence spectral data.A minimum inhibitory concentration assay were performed to evaluate the biological properties of the pure compound against multi-drug resistant pathogens.Results:Bioassay guided fractionation of the ethyl acetate extract of the culture of Salinispora sp.FS-0034 by different chromatographic methods yielded the isolation of an antibacterial compound,which was identified as rifamycin W(compound 1).Rifamycin W was reported for its potent antibacterial activity against methicillin-resistant Staphylococcus aureus,wild type Staphylococcus aureus and vancomycin-resistant Enterococcus faecium and displayed minimum inhibitory concentrations of 15.62,7.80 and 250.00 μg/mL,respectively.Conclusions:The present study reported the rifamycin W from sponge-associated Salinispora sp.and it exhibited appreciable antibacterial activity against multi-drug resistant human pathogens which indicated that sponge-associated Actinobacteria are significant sources of bioactive metabolites.

Prevalence of multi-drug resistant uropathogenic Escherichia coli in Potohar region of Pakistan

Objective:To scrutinize patterns of multi-drug-resistant uropathogenic Escherichia coli(UPEC) strains and particularly of fluoroquinolone-resistance this is an alternative choice for the treatment of urinary tract infections.Methods:Bacterial samples(n = 250) were collected from out-patients from August 2012 to August 2014 Islamabad.Antibiotic susceptibility profiling and determination of minimum inhibitory concentrations(MICs) and minimum bactericidal concentrations were performed according to the guidelines of Clinical and Laboratory Standards Institute(CLSI,2012).Genes,qnrA,qnrB and qnrS were identified by DNA amplification and sequencing.Results:The highest percentage of UPEC isolates were resistant to co-trimoxazole(82%) followed by cephalothin(80%),2nd Gen,3rd Gen and 4th Gen cephalosporins,respectively.Resistance against gentamicin,amikacin remained 29% and 4%.For other drugs including nitrofurantoin,tetracycline,carbapenem and beta-lactam inhibitors remained below 10%.Altogether,59% of the isolates were resistant to at least three antibiotics including one fluoroquinolone.Overall,MICs for ciprofloxacin remained(MIC≥256 μg/mL) and for levofloxacin(MIC≥16 μg/mL and 32 μg/mL).No significant differences were observed regarding MIC values of extended spectrumβ-lactamase(ESBL) and non-ESBL producers.For qnrS and qnrB positive isolates MICs remained above 32 μg/mL.Prevalence of UPEC was significantly higher among females and 40% of the isolates were ESBL producers.Conclusions:Higher percentages of ESBL producing UPEC were associated with urinary tract infections.Moreover,the majority of these isolates were multi-drug resistant and fluoroquinolone-resistant.

Multi-Drug Resistant Acute Otitis Media amongst Children Attending Out-Patient Clinic in Chukwuemeka Odumegwu Ojukwu University Teaching Hospital, Awka, South-East Nigeria

Objective: The study assessed microbial etiology and multi-drug resistant profile of the pathogens isolated from children diagnosed of acute otitis media (AOM) at Chukwuemeka Odumegwu Ojukwu University Teaching Hospital, Awka. Method: Ear swab samples were collected aseptically from 40 children that visited the children out-patient clinic of the hospital. The samples were cultured within 20 minutes of collection and the isolates identified microbiologically. Antimicrobial susceptibility testing was done by Kirby-Bauer’s disc diffusion method. Results: A total of 2025 children visited the hospital during the study period and 40 (1.98%) had clinical diagnosis of acute otitis media. Ten (25%) of the 40 samples did not yield any microbial growth. Children aged ≤one year had the highest prevalence of AOM (55%). Staphylococcus aureus (34.9%) was the predominant bacteria isolate followed by Pseudomonas aeruginosa (30.2%) while Candida krusei (14%) was the predominant fungi isolate. All the bacterial isolates were resistant to ampicillin and coamoxiclav. Staphylococcus aureus isolates were marginally susceptible to gentamicin (60%) and sparfloxacin (73.3%). Pseudomonas aeruginosa, Escherichia coli and Staphylococcus aureus were resistant to ceftriaxone, ciprofloxacin, streptomycin, cefuroxime, erythromycin and azithromycin. Nystatin and voriconazole had comparable effect (60%) against the fungal isolates and fluconazole was ineffective. Conclusion: The incidence of multiple antibiotic resistant bacteria associated with acute otitis media in the pediatric patient was very high. Empiric treatment of the infection in the community should be discouraged. AOM preventive strategies should be targeted on the very young children to reduce the risk of recurrence.

Bacterial contamination of orally-consumed crude herbal remedies:A potential source for multi-drug resistant pathogens in man

Objective:The acceptability of herbal remedies for alleviating discomforts and ill-health has become very popular, on the account of the increasing cost of allopathic medicine for personal health maintenance.The observable non-adherence of herbalists to the established World Health Organization(WHO) / National Agency for Food and Drug Administration Control(NAFDAC) regulations for the quality control of herbal medicines is an issue for concern.In view of this,34 popular and widely consumed crude herbal remedies in southwestern,Nigeria were screened for compliance with standard limits for bacterial contamination,bacteria flora and their antibiotic susceptibility pattern.Methods:Isolates recovered from samples were identified using the cultural, morphological and biochemical characteristics.They were also tested for drug sensitivity using standard procedures. Results:A heavy bacteria load ranging from 3.00×10~3-9.58×10~5 CFU/ML and 1.20×10~5- 5.41×10~5 CFU/ML was observed for water and spirit extracted preparations respectively.The bacteria flora cum contaminants were:Staphylococcus aureus,Bacillus cereus,Bacillus subtilis,Pseudomonas aeruginosa, Micrococcus luteus,Lactobacillus plantarum,Klebsiella pneumoniae,Escherichia coli,streptococcus,Shigella, Neisseria,Arthrobacter,Kurthia and Clostridium species.All the isolates were multi-drug resistant(MDR) strains.Conclusion:The crude herbal preparations consumed in Nigeria failed to comply with the internationally recognized standards regarding bacteria load and flora.The presence of MDR pathogens is of greatest concern. It poses a great risk to consumers health and could be a source of introducing MDR organisms into the human population.There is the need for the enforcement of established guidelines to ensure the safety of these preparations.

Management of Multi-Drug Resistant Methicillin Resistant <i>Staphylococcus aureus</i>Induced Pneumonia with New Antibiotic Adjuvant Entity: A Retrospective Study

Aim/Objective: Increase in incidences of pneumonia due to multi-drug resistant methicillin resistant Staphylococcus aureus (MRSA) in both community and health care settings is of great concern globally. Present study aims to retrospectively analyze the efficacy of new fixed dose combination with antibiotic adjuvant entity (FDC) in comparison with vancomycin to treat patients with multi-drug resistant MRSA pneumonia. Materials and Methods: During this retrospective study, case sheets of patients who were treated for MRSA pneumonia with vancomycin or fixed dose combination of vancomycin + ceftriaxone + adjuvant (FDC) between 20 March 2010 to 20 October 2014 at tertiary care center, were analyzed. Various demographic features, antibiotic therapy, length of treatment duration and the resulting efficacy were evaluated. Microbiological success was measured in terms of bacterial eradication, while clinical success was monitored in terms of complete omission of systemic signs and symptoms. Results: Among 136 patients analyzed, 113 cases were having positive culture for MRSA, and hence were further analyzed. Out of these 113 patients, empirical treatment with vancomycin was given in 59 patients and 54 patients were treated with FDC empirically. After initial culture reports, 22 patients showing resistance to vancomycin were shifted to FDC. Amidst all the patients, 24 (64.86%) of 37 from vancomycin group and 62 (81.57%) of 76 from FDC group achieved clinical success. 9 patients out of these failure cases were cured with FDC + colistin combination therapy. Failure rates in FDC treated patients were significantly low (6.57%) as compared to vancomycin group (13.51%). Conclusion: For the treatment of different types of multi-drug resistant MRSA pneumonia, the empirical intravenous FDC therapy was safe and well tolerated with higher efficacy than vancomycin. Most of the vancomycin failure cases responded to FDC therapy and were cured. This retrospective study also concludes that an alternative option of FDC + colistin is safe and effective to treat the patients which fail to respond to FDC monotherapy.