Rate Of Resistance Development Research Articles

Investigation of the Antitrichomonal Activity of Cinnamaldehyde, Carvacrol and Thymol and Synergy with Metronidazole.

Trichomonas vaginalis is a sexually transmitted protozoan parasite that usually causes infections in women. Metronidazole is used as the first choice in the treatment of this parasitic disease, but there is a need for new drugs since 1980's with increasing numbers of reported resistance. In this study, it was aimed to determine the antitrichomonal activity of the major components of Cinnamomum zeylanicum (cinnamon) and Thymus vulgaris (thyme) essential oils, cinnamaldehyde, carvacrol and thymol against metronidazole resistant and susceptible T. vaginalis strains, and to determine their interaction with metronidazole by checkerboard method. Cinnamaldehyde, carvacrol, thymol and metronidazole were obtained commercially. Two clinical isolates and one metronidazole resistant T. vaginalis reference strain were used in the study. MIC50 and MLC values of essential oil components and metronidazole were determined by broth microdilution method. The combinations of essential oil components with metronidazole were determined by the checkerboard method. According to in vitro activity tests, cinnamaldehyde was determined to be most effective essential oil component. Clinical isolates were susceptible to metronidazole. In combination study, metronidazole showed synergy with cinnamaldehyde and carvacrol, and partial synergy with thymol. It was determined that cinnamaldehyde, carvacrol and thymol, which are known to have high antimicrobial activity, also have strong activity against T. vaginalis isolates and show a synergistic interaction with metronidazole. The use of metronidazole at lower doses in the synergistic interaction may contribute to the literature in terms of reducing drug side effects, creating a versatile antimicrobial target, and reducing the rate of resistance development.

Resistance risk assessment, cross-resistance potential and realized heritability of resistance to methomyl in Musca domestica Linnaeus.

The use of insecticides in agricultural settings often exerts negative effects on nontarget species. Methomyl, a broad-spectrum carbamate insecticide, is recommended to manage a number of insect pests of the cotton crop. Recently, Musca domestica, which is a nontarget insect species in cotton fields, has shown resistance to methomyl in Pakistan. The present study tried to assess resistance-risk assessment, rapidity of resistance development to methomyl, cross-resistance potential to other insecticides, resistance heritability and to forecast the projected rate of resistance development under field conditions. For this purpose, a field strain of M. domestica with 186 fold resistance to methomyl was re-selected in the laboratory for eight consecutive generations. Consequently, LD50 values increased rapidly (126.64 ng/fly to 3112.79 ng/fly) compared to those before selection experiments. Similarly, RR values increased from 186 to 3113 fold as a result of the selection process. However, resistance to methomyl did not remain stable when the selected strain (Meth-SEL) reared for the next five generations in a pesticide free environment. The Meth-SEL strain also developed cross-resistance to permethrin. The realized heritability (h2) value for the Meth-SEL strain was 0.39 with 27% average mortality of M. domestica. Assuming the standard deviation (σp) value 0.27 and the h2 value 0.39 for eight generations of continuous exposure to methomyl, then five, seven, eight, ten and twelve generations at 90, 80, 70, 60 and 50% selection intensity, respectively, would be required for a tenfold increase in the LD50 value of methomyl. In conclusion, the Meth-SEL strain of M. domestica exhibited a high risk of resistance development to methomyl under continuous selection pressure. Resistance increased rapidly during selection experiments that reflect the probability of resistance development under field conditions if M. domestica receive exposures to methomyl during its applications for the management of target pest species.

Mathematical modelling of antibiotic interaction on evolution of antibiotic resistance: an analytical approach.

The emergence and spread of antibiotic-resistant pathogens have led to the exploration of antibiotic combinations to enhance clinical effectiveness and counter resistance development. Synergistic and antagonistic interactions between antibiotics can intensify or diminish the combined therapy's impact. Moreover, these interactions can evolve as bacteria transition from wildtype to mutant (resistant) strains. Experimental studies have shown that the antagonistically interacting antibiotics against wildtype bacteria slow down the evolution of resistance. Interestingly, other studies have shown that antibiotics that interact antagonistically against mutants accelerate resistance. However, it is unclear if the beneficial effect of antagonism in the wildtype bacteria is more critical than the detrimental effect of antagonism in the mutants. This study aims to illuminate the importance of antibiotic interactions against wildtype bacteria and mutants on the deacceleration of antimicrobial resistance. To address this, we developed and analyzed a mathematical model that explores the population dynamics of wildtype and mutant bacteria under the influence of interacting antibiotics. The model investigates the relationship between synergistic and antagonistic antibiotic interactions with respect to the growth rate of mutant bacteria acquiring resistance. Stability analysis was conducted for equilibrium points representing bacteria-free conditions, all-mutant scenarios, and coexistence of both types. Numerical simulations corroborated the analytical findings, illustrating the temporal dynamics of wildtype and mutant bacteria under different combination therapies. Our analysis provides analytical clarification and numerical validation that antibiotic interactions against wildtype bacteria exert a more significant effect on reducing the rate of resistance development than interactions against mutants. Specifically, our findings highlight the crucial role of antagonistic antibiotic interactions against wildtype bacteria in slowing the growth rate of resistant mutants. In contrast, antagonistic interactions against mutants only marginally affect resistance evolution and may even accelerate it. Our results emphasize the importance of considering the nature of antibiotic interactions against wildtype bacteria rather than mutants when aiming to slow down the acquisition of antibiotic resistance.

Susceptibility baselines for the invasive mealybugs Phenacoccus manihoti and Paracoccus marginatus (Hemiptera: Pseudococcidae) in cassava ecosystem against selected neonicotinoid insecticides

In recent years, an invasive cassava mealybug Phenacoccus manihoti has been threatening cassava cultivation alongside another invasive papaya mealybug Paracoccus marginatus which invaded the country more than a decade ago. In order to evaluate their responses against the commonly used neonicotinoid insecticides: thiamethoxam 25 WG and imidacloprid 17.8 SL, acute toxicity experiments to determine the susceptibility baselines in populations of two invasive mealybugs in the cassava agro-ecosystem, namely, cassava mealybug P. manihoti and papaya mealybug P. marginatus were performed upto 15 generations. A systemic uptake method was used for the bioassay. The LC50 values of thiamethoxam for F1 generation were 3.298 ppm whereas it was 1.066 ppm for F15 in cassava mealybug. The LC50 values of F1 generation were 2.014 ppm and that of F15 generation was 1.384 ppm when tested with imidacloprid. In the case of papaya mealybug, the LC50 values ranged from 6.138 ppm (F1) to 2.503 ppm (F15) for thiamethoxam and 7.457 ppm (F1) to 3.231 ppm (F15) for imidacloprid. All the susceptibility indices calculated were less than threefold. The rate of resistance development was negative in all cases showing that none of the tested populations harboured any resistance without insecticidal selection pressure. Tentative discriminating doses were fixed for both chemicals with the help of LC95 values obtained from the bioassay experiments, namely five ppm for both thiamethoxam and imidacloprid in the case of cassava mealybug and 10 ppm and 15 ppm, respectively, for thiamethoxam and imidacloprid in the case of papaya mealybug.

Landscape configurations of refuge areas that delay the evolution of resistance to Bt sugarcane: an agent based modeling approach.

Although transgenic crops expressing genes from the bacterium Bacillus thuringiensis (Bt) are considered to be an effective pest control method, reckless usage adds environmental pressure on a pest population to develop resistance to the protein over time. The use of small portions of non-Bt crop (refuge areas) limits the rate of resistance development. Strains of Bt sugarcane for the South African market are being developed, and a prerequisite to releasing such a product on the market is a recommendation on the size and layout of the refuge areas. In this article, an agent-based simulation model is used to test the effectiveness of different landscape configurations of refuge areas in Bt sugarcane against resistance development occurring in an associated lepidopteran pest population. Individual insects are modeled as agents on an underlying sugarcane field that can either be Bt or refugium. The model is applied to 2 hypothetical case studies, each focusing on a specific aspect of refugia planning. The first focuses on the size and distribution of refuge, and the second on the shape of the refuge. A conservative general recommendation of 30% per farm, planted in large blocks on farms, is made based on simulation results and what is currently known about the target pest species, to provide regulatory bodies as well as growers with a starting point on how to regulate and plan refuge areas in South African Bt sugarcane.

Impact of antimicrobial resistance on health and economy: A comprehensive review

Antibiotics have historically revolutionized medical science; however, the rise of multidrug-resistant pathogenic bacteria puts their worth at risk. The problem of antibiotic resistance has been attributed to both the excessive use of already available drugs and the absence of newer treatments as a result of stringent regulatory requirements and weakened corporate incentives. Resistant infections often need more comprehensive and costly treatments, resulting in longer hospital admissions, higher healthcare costs, and a larger demand for specialized healthcare resources. The expenses are exacerbated by the need for new antimicrobial drug research and development, which is becoming more complex and expensive as a result of resistance mechanisms. Comprehensive efforts are needed to limit the rate of resistance development and include research into new bacteria, resistance mechanisms, and antimicrobial medications. Multidisciplinary approaches are required in the sectors of agriculture, the environment, and healthcare. Probiotics, antibodies, and vaccines are a few examples of progressive complementary therapies that have shown promising outcomes in research and may one day serve as preventive or supplementary treatments. The purpose of this review article is to give an in-depth review of the influence of antibiotic resistance on both the health of individuals and economic stability.

Evolution of Herbicide Resistant Weeds in Agro-ecological Systems

Weed management through herbicides continues to be the most efficient and cost-effective component of integrated weed management (IWM) in crop production systems. However, the current production systems are characterised by intensive, inappropriate use of herbicides along with misapplication which creates selection pressure and leads to the rapid evolution of herbicide resistance in weeds. Currently, there are 511 unique cases of herbicide-resistant weed species globally, involving 266 species and these comprise 153 and 113 dicots and monocots respectively. To date, herbicide resistance has been reported from 96 crops in more than 71 countries. Furthermore, weeds have evolved resistance to 21 of the 31 known sites of herbicide action in different crops. Understanding the mechanisms of herbicide resistance is therefore essential which when coupled with the ecological and management factors that affect herbicide resistance would lead to the development of appropriate, profitable, and sustainable weed management strategies. The review is aimed at the development of herbicide resistance, mechanisms of herbicide resistance, factors that influence the rate of resistance development and management of herbicide resistance by growers.

Abamectin, Hexythiazox and Spiromesifen Resistance in Populations of Tetranychus urticae Koch (Acari:Tetranychidae) Collected from Cucumber Greenhouses in Tokat Province

Tetranychus urticae Koch (Acari: Tetranychidae) is one of the most important pests in greenhouses. This harmful pest has rapidly developed high resistance to many classes of acaricides. In this study, T. urticae populations were collected from cucumber greenhouses in Gümenek village, Tokat, Turkey. Growers in this location use acaricides such as abamectin, hexythiazox and spiromesifen to suppress T. urticae populations. The study aimed to determine the resistance levels of T. urticae populations against to mentioned acaricides. The LC50 values of the collected populations were determined by the spray tower-leaf disc method. The resistance rates were found by dividing the LC50 values of the collected greenhouse populations by the LC50 value of the susceptible population. The resistance levels of T. urticae were determined to be between 1.88-2.14-folds against abamectin, 1.67-1.84 folds against hexythiazox and 1.77-2.09 folds against spiromesifen. According to these results, a low rate of resistance development was observed against abamectin, hexythiazox and spiromesifen.

Segregation of Cry Genes in the Seeds Produced by F1 Bollgard® II Cotton Differs between Hybrids: Could This Be Linked to the Observed Field Resistance in the Pink Bollworm?

Indian populations of the Pink Bollworm (PBW) are resistant to Bt (Bacillus thuringiensis) cotton hybrids containing Cry1Ac and Cry2Ab genes. Segregation of these Cry genes in F1 hybrids could subject PBW to sublethal concentrations. Moreover, planting hybrids with varying zygosities of Cry genes could produce diverse segregation patterns and expose PBW populations to highly variable toxin concentrations. This could potentially promote the rate of resistance development. Therefore, we studied the segregation patterns of Cry genes in different commercial Bt hybrids cultivated in India. Results showed that two hybrids segregated according to the Mendelian mono-hybrid ratio, three segregated according to the Mendelian di-hybrid ratio, and one showed a mixed segregation pattern. The assortment of seeds containing Cry genes varied between bolls of the same hybrid. In India, different Bt cotton hybrids are cultivated in small patches next to each other, exposing PBW populations to sublethal doses and wide variations in the occurrence of Cry genes. It is necessary to avoid segregation of Cry genes in the seeds produced by F1 hybrids. This study recommends using Bt parents homozygous for Cry genes in commercial Bt cotton hybrid development. This breeding strategy could be effective for similar transgenic crop hybrids as well.

Assessing the resistance risk to emamectin benzoate in Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae): selection, cross-resistance dynamics and estimates of realized heritability

• 280.15-fold resistance was detected in Phenacoccus solenopsis to emamectin benzoate. • CR to abamectin and profenofos was 0.28-1.65-fold, and 0.12-0.29-fold, respectively. • CR to cypermethrin ranged from 6.02-11.29-fold. • h 2 of resistance to emamectin benzoate was 0.24 from G 3 to G 19. Emamectin benzoate, a non-systemic, translaminar, and broad spectrum insecticide, is excessively used in cotton. It kills the pest by suppressing the muscle contraction. Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) is one of the major sap sucking pests of cotton in many countries and has been shown resistance to various insecticides. In this study, we have continuously selected P. solenopsis with emamectin benzoate in the laboratory from G 3 to G 18 to determine the resistance development and resistance risk to emamectin benzoate and also to evaluate the dynamics of cross-resistance (CR) development with three other insecticides. After continuing selection of P. solenopsis for 16 generations with emamectin benzoate, the emamectin selected strain (Ema-SEL) developed a 280.15-fold resistance when tested at G 19 . Ema-SEL strain showed no CR to abamectin (0.28-1.65-fold), and profenofos (0.12-0.29-fold), but a very low to low CR to cypermethrin (6.02-11.29-fold). Abamectin was negatively cross-resistant with emamectin benzoate in Ema-SEL strain at G 13 , G 15 and G 19 while profenofos also exhibited similar results at G 13 , G 15 , G 17 and G 19 . Realized heritability ( h 2 ) of resistance to emamectin benzoate was 0.24 from G 3 to G 19 . The results of projected rate of resistance development showed that if h 2 = 0.24, 0.34, and 0.44 at slope = 1.08, then 2.03, 1.43, and 1.11 generations, respectively are required for ten-fold increase in LC 50 at 95% selection intensity. Results of the present study suggested that P. solenopsis has the ability to develop resistance to emamectin benzoate but it can be managed by the alternate use of other insecticides including abamectin, profenofos and cypermethrin due to lack of/or very low to low CR with these insecticides.

Animal experimental investigation on the efficacy of antibiotic therapy with linezolid, vancomycin, cotrimoxazole, and rifampin in treatment of periprosthetic knee joint infections by MRSA.

AimsPeriprosthetic joint infections (PJIs) are rare, but represent a great burden for the patient. In addition, the incidence of methicillin-resistant Staphylococcus aureus (MRSA) is increasing. The aim of this rat experiment was therefore to compare the antibiotics commonly used in the treatment of PJIs caused by MRSA.MethodsFor this purpose, sterilized steel implants were implanted into the femur of 77 rats. The metal devices were inoculated with suspensions of two different MRSA strains. The animals were divided into groups and treated with vancomycin, linezolid, cotrimoxazole, or rifampin as monotherapy, or with combination of antibiotics over a period of 14 days. After a two-day antibiotic-free interval, the implant was explanted, and bone, muscle, and periarticular tissue were microbiologically analyzed.ResultsVancomycin and linezolid were able to significantly (p < 0.05) reduce the MRSA bacterial count at implants. No significant effect was found at the bone. Rifampin was the only monotherapy that significantly reduced the bacterial count on implant and bone. The combination with vancomycin or linezolid showed significant efficacy. Treatment with cotrimoxazole alone did not achieve a significant bacterial count reduction. The combination of linezolid plus rifampin was significantly more effective on implant and bone than the control group in both trials.ConclusionAlthough rifampicin is effective as a monotherapy, it should not be used because of the high rate of resistance development. Our animal experiments showed the great importance of combination antibiotic therapies. In the future, investigations with higher case numbers, varied bacterial concentrations, and changes in individual drug dosages will be necessary to be able to draw an exact comparison, possibly within a clinical trial.Cite this article: Bone Joint Res 2022;11(3):143–151.

Reversion of CYP450 monooxygenase-mediated acetamiprid larval resistance in dengue fever mosquito, Aedes aegypti L.

Aedes-borne diseases are on the rampant rise despite continued application of chemical insecticide-based interventions. The appearance of high degree of insecticide resistance in Aedes species and noxious effects on environment and non-targets have raised further concerns. Among new chemical interventions, neonicotinoids are considered a safe and effective approach. The present study investigated the control potency of acetamiprid and development of resistance in Aedes aegypti larvae; and the involvement of CYP450 monooxygenases in inducing resistance. The early fourth instars of Ae. aegypti parent susceptible strain (PS) were selected with acetamiprid for 15 generations (ACSF strain) increasing the resistance to 19.74-fold in ACSF-10 and 36.71-fold in ACSF-15. The ACSF-10 larvae were assayed with acetamiprid combined with piperonyl butoxide (PBO) in three different ratios (1:1, 1:5 and 1:10) and selected for next five generations with 1:10 combination. Selection with synergized acetamiprid (APSF strains) reversed as well as reduced the rate of resistance development resulting in only 1.35-fold resistance in APSF-15. The APSF strains showed %monooxygenase dependency ranging from 86.71 to 96.72%. The estimation of the monooxygenases levels in parent and selected larvae showed increased monooxygenase level in the ACSF strains by 2.42-2.87-fold. The APSF-15 strains exhibited 57.95% lower enzyme production than ACSF-15 strain. The reduction and reversion of resistance by using PBO and the elevated levels of monooxygenases in ACSF and reduction in APSF strains recommend the involvement of CYP450-mediated mechanism in the development of acetamiprid resistance in Ae. aegypti. These studies could help in devising resistance management strategies in order to preserve the efficiency of pre-existing insecticides.

Risk assessment of cyromazine and methoxyfenozide resistance suggests higher additive genetic but lower environmental variation supporting quick resistance development in non-target Chrysoperla carnea (Stephens).

Insecticides are effective against economic pests, but these pose serious threats to the environment and ecosystem components such as natural enemies. Resistance risk assessment forecasts insecticide resistance development in target pests and non-target biological control agents under special conditions. Field-collected Chrysoperla carnea was selected with two Insect Growth Regulators (IGRs) viz. cyromazine and methoxyfenozide for 15 generations to determine the resistance development potential of this natural enemy. Selection to cyromazine and methoxyfenozide induced 759.08-fold and 3531.67-fold resistance with realized heritability of 0.37 and 0.62 in C. carnea, respectively, suggesting higher additive genetic variations in first half of selection (h2 = 0.46 for cyromazine and h2 = 0.75 for methoxyfenozide) than in second half (h2 = 0.18 and 0.25, respectively). Estimates of projected rate of resistance development indicate C. carnea will take only 6 to 2 generations at h2 = 0.37, 8 to 2 at h2 = 0.27, and 5 to 2 at h2 = 0.27, at constant slope = 1.81 for a tenfold increase in cyromazine resistance. At h2 = 0.37, 3-1, and 10-8 generations would be needed for this increase in LC50 if slope = 0.82 and 2.82, respectively. Similarly, it may take 3 to 1 generations at h2 = 0.62 and 0.72, but 4 to 1 at h2 = 0.52, at constant slope = 1.62, for a tenfold increase in methoxyfenozide resistance. On the same h2 = 0.62, 1-0, and 5-1 generations would be required for increase if slope = 0.62 and 2.62, respectively. Selection and resistance to both insecticides induced an insignificant difference in the sex ratio of C. carnea. These results confirm that this natural enemy has tremendous potential for resistance development under selection pressure.

Retracted: Iron Oxide Nanoparticles: Preparation, Characterization, and Assessment of Antimicrobial and Anticancer Activity

Nanotechnology and nanoparticles (NPs) have increasingly been studied as an alternative for antibiotics because of the feasibility to be used in implantable devices both for bacterial detection and infection prevention. The low rate of resistance development against NPs because of its multiple mode of action has contributed to its increased acceptance in clinical setting. Further development of NPs and their anticancer activity against many human cancer cell lines including breast and ovarian have been documented. Fe2O3-NPs could be used for antibacterial and anticancer activity assessment. Iron oxide, apart from being available extensively and cheap, also plays a role in multiple biological processes, making it an interesting metal for NPs. The aim of the present study revolves around generation and characterization of iron oxide Fe2O3-NPs, followed by assessment of its antimicrobial and anticancer activities. Synthesis of Fe2O3-NPs was performed by hydrothermal approach, and its characterization was done by UV-visible, X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR) analyses, and transmission electron microscopy (TEM). Antimicrobial activity was checked by agar diffusion assay against Bacillus subtilis, Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, and Candida albicans. Anticancer activity of the NPs was assessed using the human cancer cell lines including cervical carcinoma cell line (HeLa) and MCF7. The developed Fe2O3-NPs exhibited a characteristic absorption curve in the 500-600 nm wavelength range by UV-visible analysis, the XRD peaks were found to index the rhombohedral shape, and the FTIR analysis ascertained the bonds and functional groups at wavenumber from 400 to 4000 cm-1. Antimicrobial assay detected significant effect against Staphylococcus aureus and Bacillus subtilis with zones of inhibition: 21 and 22 mm, respectively. Likewise, Fe2O3-NPs show good activity towards tested fungal strain Candida albicans with zone of inhibition of 24 mm. The 2,5-diphenyl-2H-tetrazolium bromide (MTT) assay identified significant anticancer activity of the NPs against both cell lines. Our study documents the successful generation and characterization of Fe2O3-NPs having excellent antimicrobial and anticancer activities.

Resistance status and the rate of resistance development in Musca domestica Linnaeus (Diptera: Muscidae) to permethrin and imidacloprid in Indonesia

A previous report has indicated that in many regions of Indonesia, populations of Musca domestica Linnaeus have shown very high resistance to permethrin and low resistance to imidacloprid. In this study, the resistance status to permethrin and imidacloprid was updated using a topical application and feeding bioassay. Six housefly strains originated from six highly populated cities in Indonesia, namely Serang (SRG), Jakarta (JKT), Bandung (BDG), Semarang (SMG), Yogyakarta (JOG), and Surabaya (SBY). A seventh strain (Danish Pest Infestation Laboratory (DPIL)) served as the control. Each strain was tested for resistance to the two insecticides. In addition, the rate of development of resistance to the two insecticides was measured over ten generations. The results indicated that all field strains showed very high resistance to permethrin. The highest resistance level recorded was in the SRG strain (RR50 = 2880), and the lowest was in the JKT strain (RR50 = 520). Repeated application of permethrin over ten generations increased the resistance level by about 2.7–32.73-fold as compared to the level of their respective parental populations. On the other hand, most strains showed low to moderate resistance to imidacloprid, in which the SRG strain had the highest resistance level (RR50 = 15.5) and the SBY strain had the lowest (RR50 = 2.0). Repeated application of imidacloprid over ten generations increased the resistance level by about 3.25–17.41-fold. The findings, which is the second report of housefly resistance in Indonesia since 2016, provide a crucial foundation for developing appropriate housefly integrated pest management strategies in highly populated areas in Indonesia.

Laboratory Selection and Assessment of Resistance Risk in Drosophila suzukii (Diptera: Drosophilidae) to Spinosad and Malathion.

Simple SummaryContinuous insecticide applications used to prevent fruit infestations by spotted-wing drosophila (SWD), an invasive pest of soft-skinned fruits worldwide, can elevate the risk of resistance development in D. Suzukii field populations. However, proactive assessment of resistance risk using laboratory selection provides valuable information for development of sustainable resistance management strategies for SWD. After 10 and 11 generations of artificial selection of a colony of field-collected SWD for resistance against spinosad and malathion, a 7.55- and 2.23-fold resistance to spinosad and malathion was realized. A quantitative genetic approach used to estimate realized heritability (h2) of resistance shows that the risk of resistance in SWD populations exists against both spinosad and malathion, and a faster rate of resistance development is expected against spinosad. However, timely implementation of resistance management strategies can slow the development of resistance and prolong effective life of these insecticides against D. suzukii. Drosophila suzukii (Matsumura) is one of the most economically important pests of soft-skinned fruits worldwide. Repeated insecticide applications commonly used to prevent fruit infestations increase the risk of resistance development in D. suzukii. Assessment of resistance risk in D. suzukii using artificial selection can be valuable in developing proactive resistance management strategies to retain susceptibility in the field populations. Here, we artificially selected a colony of field-collected D. suzukii for resistance against spinosad and malathion. A quantitative genetic approach was then used to estimate realized heritability (h2) of resistance and predict the rates of resistance development. After 10 and 11 generations of selection, resistance to spinosad and malathion in D. suzukii females significantly increased by 7.55- and 2.23-fold, respectively. Based on the predicted rates of resistance development, assuming h2 = 0.14 (mean h2 of spinosad resistance in this study) and 90% of population was killed at each generation, 10-fold increase in LC50 of D. suzukii females would be expected in nine generations for spinosad. However, 10-fold increase in LC50 of D. suzukii females for malathion would be expected in 37 generations, assuming h2 = 0.08 (mean h2 of malathion resistance) and 90% of population was killed at each generation. These results indicate that the risk of resistance in D. suzukii populations exists against both spinosad and malathion. However, resistance would develop faster against spinosad as compared to malathion. Thus, resistance management strategies should be implemented proactively to maintain the effectiveness of these insecticides to control D. suzukii.

Lefamulin: A Novel Oral and Intravenous Pleuromutilin for the Treatment of Community-Acquired Bacterial Pneumonia.

Lefamulin is a novel oral and intravenous (IV) pleuromutilin developed as a twice-daily treatment for community-acquired bacterial pneumonia (CABP). It is a semi-synthetic pleuromutilin with a chemical structure that contains a tricyclic core of five-, six-, and eight-membered rings and a 2-(4-amino-2-hydroxycyclohexyl)sulfanylacetate side chain extending from C14 of the tricyclic core. Lefamulin inhibits bacterial protein synthesis by binding to the 50S bacterial ribosomal subunit in the peptidyl transferase center (PTC). The pleuromutilin tricyclic core binds to a pocket close to the A site, while the C14 side chain extends to the P site causing a tightening of the rotational movement in the binding pocket referred to as an induced-fit mechanism. Lefamulin displays broad-spectrum antibacterial activity against Gram-positive and Gram-negative aerobic and anaerobic bacteria as well as against atypical bacteria that commonly cause CABP. Pleuromutilin antibiotics exhibit low rates of resistance development and lack cross-resistance to other antimicrobial classes due to their unique mechanism of action. However, pleuromutilin activity is affected by mutations in 23S rRNA, 50S ribosomal subunit proteins rplC and rplD, ATP-binding cassette (ABC)-F transporter proteins such as vga(A), and the methyltransferase cfr. The pharmacokinetic properties of lefamulin include: volume of distribution (Vd) ranging from 82.9 to 202.8 L, total clearance (CLT) of 19.5 to 21.4 L/h, and terminal elimination half-life (t1/2) of 6.9-13.2 h; protein binding of lefamulin is high and non-linear. The oral bioavailability of lefamulin has been estimated as 24% in fasted subjects and 19% in fed subjects. A single oral dose of lefamulin 600 mg administered in fasted patients achieved a maximum plasma concentration (Cmax) of 1.2-1.5 mg/L with a time of maximum concentration (Tmax) ranging from 0.8 to 1.8 h, and an area under the plasma concentration-time curve from 0 to infinity (AUC0-∞) of 8.5-8.8 mgh/L. The pharmacodynamic parameter predictive of lefamulin efficacy is the free plasma area under the concentration-time curve divided by the minimum inhibitory concentration (fAUC24h/MIC). Lefamulin efficacy has been demonstrated using various animal models including neutropenic murine thigh infection, pneumonia, lung infection, and bacteremia. Lefamulin clinical safety and efficacy was investigated through a Phase II clinical trial of acute bacterial skin and skin structure infection (ABSSSI), as well as two Phase III clinical trials of CABP. The Phase III trials, LEAP 1 and LEAP 2 established non-inferiority of lefamulin to moxifloxacin in both oral and IV formulations in the treatment of CABP. The United States Food and Drug Administration (FDA), European Medicines Agency (EMA), and Health Canada have each approved lefamulin for the treatment of CABP. A Phase II clinical trial has been completed for the treatment of ABSSSI, while the pediatric program is in Phase I. The most common adverse effects of lefamulin include mild-to-moderate gastrointestinal-related events such as nausea and diarrhea. Lefamulin represents a safe and effective option for treating CABP in cases of antimicrobial resistance to first-line therapies, clinical failure, or intolerance/adverse effects to currently used agents. Clinical experience and ongoing clinical investigation will allow clinicians and antimicrobial stewardship programs to optimally use lefamulin in the treatment of CABP.

Targeting pathogenic fungi, bacteria and fungal-bacterial biofilms by newly synthesized quaternary ammonium derivative of pyridoxine and terbinafine with dual action profile

Many pathogenic bacteria and microscopic fungi form rigid polymicrobial biofilms this way enhancing their resistant to treatment. A series of novel pyridoxine-based quaternary ammonium derivatives of terbinafine characterized by both antifungal and antibacterial activities was designed. The leading compound named KFU-127 exhibits promising antifungal and antibacterial activities against various bacteria and micromycetes in both planktonic and biofilm-embedded forms demonstrating MIC values comparable with those of conventional antifungals and antimicrobials. Similar to other antiseptics like benzalkonium chloride and miramistin, KFU-127 is considerably toxic for eukaryotic cells that limits is application to topical treatment options. On the other hand, KFU-127 reduces the number of viable biofilm-embedded bacteria and C. albicans by 3 orders of magnitude at concentrations 2–4 times lower than those of reference drugs and successfully eradicates S. aureus-C. albicans mixed biofilms. The mechanism of antimicrobial action of KFU-127 is bimodal including both membrane integrity damage and pyridoxal-dependent enzymes targeting. We expect that this bilateral mechanism would result in lower rates of resistance development in both fungal and bacterial pathogens. Taken together, our data suggest KFU-127 as a new promising broad spectrum topical antimicrobial capable of one-shot targeting of bacterial and fungal-bacterial biofilms.

Mining for novel antibiotics in the age of antimicrobial resistance.

The misuse of antimicrobials is causing an alarming increase in the appearance of antibiotic-resistant microorganisms. In this context, the identification of novel antibiotics against new targets and with low rates of resistance development is a major global challenge. In this article, we highlight a number of recent articles that exploit a variety of in vitro, in vivo and in silico state-of-the-art approaches to identify and develop new antimicrobials. Rapid progress in this research field will be crucial to combating a global health problem, antimicrobial resistance, that is expected to be the leading cause of death by 2050.

Selection of cypermethrin resistance in mosquito Culex pipiens larvae in Assiut, Egypt

To induce cypermethrinresistance in Culex pipiens larvae, about 20000 4th instar larvae of Cx. pipiens were collected from Assiut field and exposed to cypermethrin selection pressure for fifteen generations using dipping method. This study revealed that the resistance ratio (RR50) of the parent strain (G0) was 14.49-fold as compared with susceptible strain. This RR50 value increased gradually from each generation to the next one as a result of selection pressure with cypermethrin. The regression line for cypermethrin against the parent field strain showed slope value 2.38 which indicate a moderate level of heterogeneity of parent strain to cypermethrin. The first five generations during the course of cypermethrin selection showed that resistance ratio (RR50) was increased gradually from one generation to the next (The RR50 increased from 14.49-fold in G0 to 135.40-fold resistance in G5). By the 6th generation, the rate of resistance development to cypermethrin was fairly rapid where; the RR50 to cypermethrin increased from 135.40-fold in G6 to 272.00-fold resistance in G11. With the beginning of the 12th generation, the development of resistance to cypermethrin was increased very slowly during the last four generations from 279.92-fold in G12 to 295.52-fold resistance in G15 (plateau phase). Finally, the slope values were volatile until the eleventh generation (2.38 – 3.63) and then relatively proven in the last four generations (4.55 – 4.52). This is evidence of increasing homogeneity of selected strain in the last five generations.