Novel isolates of Cydia pomonella granulovirus (CpGV): deciphering the molecular mechanism for overcoming CpGV resistance in codling moth (Cydia pomonella)

During the last twenty years, Cydia pomonolla granulovirus (CpGV, Baculoviridae) has become the most important biological control agent for the codling moth (CM) in organic and integrated apple production. All registered products in Europe are based on the isolate CpGV-M, which was discovered 1964 in Mexico. A serious threat to future application of CpGV is the occurrence of CM field populations resistant to CpGV. Since 2003, populations with up to 10,000-fold reduced susceptibility were reported from orchards in Germany, France, Italy, Switzerland, Austria and the Netherlands. A putative alternative to CpGV-M are novel CpGV isolates which are able to overcome CM resistance. This thesis focuses on the identification and characterisation of resistance overcoming CpGV isolates and the analysis of their molecular difference to CpGV-M.rnSixteen CpGV isolates were tested against CM lab strains in bioassays. Hereby, five isolates were identified which were able to completely overcome resistance. The genomes of these isolates were compared to CpGV-M by restriction fragment length polymorphism (RFLP) analysis. To identify the molecular factor responsible for improved virulence of some CpGV isolates, major genomic differences were sequenced and analysed. A 0.7 kb insertion was found in CpGV-I01, -I12 and -E2, but not in other resistance overcoming isolates. Analysis of the insertions sequence revealed that it might be due to a transposition event, but not involved in overcoming resistance. rnFor unequivocal identification of CpGV isolates, a new method based on molecular analysis was established. Partial sequencing of the conserved polyhedrin/granulin (polh/gran), late expression factor-8 (lef-8) and late expression factor-9 (lef-9) genes revealed single nucleotide polymorphisms (SNPs). SNP analysis correlated with the grouping obtained by RFLP analysis. A phylogenetic classification due to different genome types A-E is proposed. Phylogenetic analysis suggested that CpGV-M was the phylogenetically youngest of the tested CpGV isolates.rnWhole genome sequencing of two resistance overcoming isolates CpGV-I12 (type D genome) and -S (type E genome) and CpGV-M (type A genome) was performed. Comparison of the three genomes revealed a high sequence identity. Several insertions and deletions ranging from 1-700 nucleotides (nt) were found. Comparison on open reading frame (ORF) level revealed that CpGV-I12 and -S shared only one protein alteration when compared to CpGV-M: a stretch of 24 nt present in ORF cp24 was not found in any of the resistance overcoming isolates. Cp24 codes for the early gene pe38. Combined with the results of phylogenetic analysis, it is proposed that these 24 nt are a recent insertion into the CpGV-M genome. The role of pe38 in overcoming resistance was investigated by knocking out pe38 of a CpGV-M based bacmid and swapping of CpGV-I12 pe38 of into the k.o. bacmid. When pe38 of CpGV-I12 was inserted into the k.o. bacmid, the infectivity could not be rescued, suggesting that the genomic portion of pe38 might play a role in its function.rnIt can be concluded that the recently observed CpGV resistance in CM is only related to type A genomes. RFLP and SNP analysis provide tools for identifying and characterising different CpGV isolates reliably, a pre-condition for a future registration of CpGV products based on novel CpGV isolates.rnrnrn

Incidence of HIV-1 drug resistance among antiretroviral treatment-naive individuals starting modern therapy combinations

Estimates of drug resistance incidence to modern first-line combination antiretroviral therapies against human immunodeficiency virus (HIV) type 1 are complicated by limited availability of genotypic drug resistance tests (GRTs) and uncertain timing of resistance emergence.

Active surveillance of antibiotic resistance prevalence in urinary tract and skin infections in the outpatient setting

The aim of the study was to evaluate the need for active surveillance of antibiotic resistance in ambulatory infections. We measured the prevalence of antibiotic resistance in urinary tract infections (UTIs) (n = 1018) and skin infections (n = 213) diagnosed in outpatients between September 2008 and February 2009 in the Canton of Bern, Switzerland. Samples were stratified into 'solicited' (diagnostic work-up for study purpose only) and 'routine' (diagnostic work-up as part of standard care). Susceptibility patterns were compared for 463 Escherichia coli isolates from UTIs (231 solicited; 232 routine) and 87 Staphylococcus aureus isolates from skin infections (35 solicited; 52 routine). Overall, E. coli showed higher susceptibility to ampicillin, amoxicillin-clavulanic acid and norfloxacin in solicited than in routine samples. Among 15-45-year-old patients, susceptibility rates were comparable between solicited and routine samples for all antibiotics except for amoxicillin-clavulanic acid. However, among patients >45 years old, isolates from routine samples showed lower susceptibility to all β-lactams tested and quinolones than those from solicited samples. Extended-spectrum β-lactamase (ESBL)-producing E. coli isolates were rare (solicited, 0.4%; routine, 1.7%; p 0.4). Susceptibility patterns of S. aureus were comparable between solicited and routine samples. Therefore, in the outpatient setting, susceptibility rates for E. coli isolates differ by indication for urinary culture and age. Surveillance based on samples taken during standard care may underestimate susceptibility rates for uncomplicated infections, especially among the elderly. Reports of resistance data should include age stratification.

Impact of antibiotic use on carbapenem resistance in Pseudomonas aeruginosa: is there a role for antibiotic diversity?

In this study, we aimed to evaluate the relationship between the rates of resistance of Pseudomonas aeruginosa to carbapenems and the levels and diversity of antibiotic consumption. Data were retrospectively collected from 20 acute care hospitals across 3 regions of Switzerland between 2006 and 2010. The main outcome of the present study was the rate of resistance to carbapenems among P. aeruginosa. Putative predictors included the total antibiotic consumption and carbapenem consumption in defined daily doses per 100 bed days, the proportion of very broad-spectrum antibiotics used, and the Peterson index. The present study confirmed a correlation between carbapenem use and carbapenem resistance rates at the hospital and regional levels. The impact of diversifying the range of antibiotics used against P. aeruginosa resistance was suggested by (i) a positive correlation in multivariate analysis between the above-mentioned resistance and the proportion of consumed antibiotics having a very broad spectrum of activity (coefficient = 1.77; 95% confidence interval, 0.58 to 2.96; P < 0.01) and (ii) a negative correlation between the resistance and diversity of antibiotic use as measured by the Peterson homogeneity index (coefficient = -0.52; P < 0.05). We conclude that promoting heterogeneity plus parsimony in the use of antibiotics appears to be a valuable strategy for minimizing the spread of carbapenem resistance in P. aeruginosa in hospitals.

Presence of new mecA and mph(C) variants conferring antibiotic resistance in Staphylococcus spp. isolated from the skin of horses before and after clinic admission

Because of the frequency of multiple antibiotic resistance, Staphylococcus species often represent a challenge in incisional infections of horses undergoing colic surgery. To investigate the evolution of antibiotic resistance patterns before and after preventative peri- and postoperative penicillin treatment, staphylococci were isolated from skin and wound samples at different times during hospitalization. Most staphylococci were normal skin commensals and belonged to the common coagulase-negative group. In some cases they turned out to be opportunistic pathogens present in wound infections. MICs were determined for 12 antibiotics, and antibiotic resistance genes were detected by microarray. At hospital admission, horses harbored staphylococci that were susceptible to antibiotics or resistant to one group of drugs, mainly due to the presence of new variants of the methicillin and macrolide resistance genes mecA and mph(C), respectively. After 3 days, the percentage of Staphylococcus isolates displaying antibiotic resistance, as well as the number of resistance genes per isolate, increased moderately in hospitalized horses without surgery or penicillin treatment but dramatically in hospitalized horses after colic surgery as well as penicillin treatment. Staphylococcus species displaying multiple resistance were found to harbor mainly genes conferring resistance to beta-lactams (mecA and blaZ), aminoglycosides [str and aac(6')-Ie-aph(2')-Ia], and trimethoprim [dfr(A) and dfr(D)]. Additional genes conferring resistance to macrolides [mph(C), erm(C), and erm(B)], tetracycline [tet(K) and tet(M)], chloramphenicol [cat(pC221) and cat(pC223)], and streptothricin (sat4) appeared in several strains. Hospitalization and preventive penicillin use were shown to act as selection agents for multidrug-resistant commensal staphylococcal flora.

Loss of the high-affinity pentamidine transporter is responsible for high levels of cross-resistance between arsenical and diamidine drugs in African trypanosomes

Treatment of many infectious diseases is under threat from drug resistance. Understanding the mechanisms of resistance is as high a priority as the development of new drugs. We have investigated the basis for cross-resistance between the diamidine and melaminophenyl arsenical classes of drugs in African trypanosomes. We induced high levels of pentamidine resistance in a line without the tbat1 gene that encodes the P2 transporter previously implicated in drug uptake. We isolated independent clones that displayed very considerable cross-resistance with melarsen oxide but not phenylarsine oxide and reduced uptake of [(3)H]pentamidine. In particular, the high-affinity pentamidine transport (HAPT1) activity was absent in the pentamidine-adapted lines, whereas the low affinity pentamidine transport (LAPT1) activity was unchanged. The parental tbat1(-/-) line was sensitive to lysis by melarsen oxide, and this process was inhibited by low concentrations of pentamidine, indicating the involvement of HAPT1. This pentamidine-inhibitable lysis was absent in the adapted line KO-B48. Likewise, uptake of the fluorescent diamidine 4',6-diamidino-2-phenylindole dihydrochloride was much delayed in live KO-B48 cells and insensitive to competition with up to 10 muM pentamidine. No overexpression of the Trypanosoma brucei brucei ATP-binding cassette transporter TbMRPA could be detected in KO-B48. We also show that a laboratory line of Trypanosoma brucei gambiense, adapted to high levels of resistance for the melaminophenyl arsenical drug melarsamine hydrochloride (Cymelarsan), had similarly lost TbAT1 and HAPT1 activity while retaining LAPT1 activity. It seems therefore that selection for resistance to either pentamidine or arsenical drugs can result in a similar phenotype of reduced drug accumulation, explaining the occurrence of cross-resistance.

Long-term trends of HIV type 1 drug resistance prevalence among antiretroviral treatment-experienced patients in Switzerland

BACKGROUND:Accurate quantification of the prevalence of human immunodeficiency virus type 1 (HIV-1) drug resistance in patients who are receiving antiretroviral therapy (ART) is difficult, and results from previous studies vary. We attempted to assess the prevalence and dynamics of resistance in a highly representative patient cohort from Switzerland. METHODS:On the basis of genotypic resistance test results and clinical data, we grouped patients according to their risk of harboring resistant viruses. Estimates of resistance prevalence were calculated on the basis of either the proportion of individuals with a virologic failure or confirmed drug resistance (lower estimate) or the frequency-weighted average of risk group-specific probabilities for the presence of drug resistance mutations (upper estimate). RESULTS:Lower and upper estimates of drug resistance prevalence in 8064 ART-exposed patients were 50% and 57% in 1999 and 37% and 45% in 2007, respectively. This decrease was driven by 2 mechanisms: loss to follow-up or death of high-risk patients exposed to mono- or dual-nucleoside reverse-transcriptase inhibitor therapy (lower estimates range from 72% to 75%) and continued enrollment of low-risk patients who were taking combination ART containing boosted protease inhibitors or nonnucleoside reverse-transcriptase inhibitors as first-line therapy (lower estimates range from 7% to 12%). A subset of 4184 participants (52%) had >or= 1 study visit per year during 2002-2007. In this subset, lower and upper estimates increased from 45% to 49% and from 52% to 55%, respectively. Yearly increases in prevalence were becoming smaller in later years. CONCLUSIONS:Contrary to earlier predictions, in situations of free access to drugs, close monitoring, and rapid introduction of new potent therapies, the emergence of drug-resistant viruses can be minimized at the population level. Moreover, this study demonstrates the necessity of interpreting time trends in the context of evolving cohort populations.

Molecular epidemiology and genetic linkage of macrolide and aminoglycoside resistance in Staphylococcus intermedius of canine origin

A collection of 77 Staphylococcus intermedius isolates from dogs and cats in Switzerland was examined for resistance to erythromycin. Resistance profiles for 14 additional antibiotics were compared between erythromycin-resistant and susceptible isolates. A resistance prevalence of 27% for erythromycin was observed in the population under study. Complete correlation between resistance to erythromycin, and to spiramycin, streptomycin, and neomycin was observed. The erythromycin-resistant isolates all had a reduced susceptibility to clindamycin when compared to the erythromycin-susceptible isolates. Both constitutive and inducible resistance phenotypes were observed for clindamycin. Ribotyping showed that macrolide-aminoglycoside resistance was randomly distributed among unrelated strains. This suggests that this particular resistance profile is not related to a single bacterial clone but to the horizontal transfer of resistance gene clusters in S. intermedius populations. The erythromycin-resistant isolates were all carrying erm(B), but not erm(A), erm(C), or msr(A). The erm(B) gene was physically linked to Tn5405-like elements known as resistance determinants for streptomycin, streptothricin, neomycin and kanamycin. Analysis of the region flanking erm(B) showed the presence of two different groups of erm(B)-Tn5405-like elements in the S. intermedius population examined and of elements found in Gram-positive species other than staphylococci. This strongly suggests that erm(B) or the whole erm(B)-Tn5405-like elements in S. intermedius originate from other bacterial species, possibly from enterococci.

Adherence as a Predictor of the Development of Class-Specific Resistance Mutations: The Swiss HIV Cohort Study

Background Non-adherence is one of the strongest predictors of therapeutic failure in HIV-positive patients. Virologic failure with subsequent emergence of resistance reduces future treatment options and long-term clinical success. Methods Prospective observational cohort study including patients starting new class of antiretroviral therapy (ART) between 2003 and 2010. Participants were naïve to ART class and completed ≥1 adherence questionnaire prior to resistance testing. Outcomes were development of any IAS-USA, class-specific, or M184V mutations. Associations between adherence and resistance were estimated using logistic regression models stratified by ART class. Results Of 314 included individuals, 162 started NNRTI and 152 a PI/r regimen. Adherence was similar between groups with 85% reporting adherence ≥95%. Number of new mutations increased with increasing non-adherence. In NNRTI group, multivariable models indicated a significant linear association in odds of developing IAS-USA (odds ratio (OR) 1.66, 95% confidence interval (CI): 1.04-2.67) or class-specific (OR 1.65, 95% CI: 1.00-2.70) mutations. Levels of drug resistance were considerably lower in PI/r group and adherence was only significantly associated with M184V mutations (OR 8.38, 95% CI: 1.26-55.70). Adherence was significantly associated with HIV RNA in PI/r but not NNRTI regimens. Conclusion Therapies containing PI/r appear more forgiving to incomplete adherence compared with NNRTI regimens, which allow higher levels of resistance, even with adherence above 95%. However, in failing PI/r regimens good adherence may prevent accumulation of further resistance mutations and therefore help to preserve future drug options. In contrast, adherence levels have little impact on NNRTI treatments once the first mutations have emerged.

Empirical chemosensitivity testing in a spheroid model of ovarian cancer using a microfluidics-based multiplex platform.

The use of biomarkers to infer drug response in patients is being actively pursued, yet significant challenges with this approach, including the complicated interconnection of pathways, have limited its application. Direct empirical testing of tumor sensitivity would arguably provide a more reliable predictive value, although it has garnered little attention largely due to the technical difficulties associated with this approach. We hypothesize that the application of recently developed microtechnologies, coupled to more complex 3-dimensional cell cultures, could provide a model to address some of these issues. As a proof of concept, we developed a microfluidic device where spheroids of the serous epithelial ovarian cancer cell line TOV112D are entrapped and assayed for their chemoresponse to carboplatin and paclitaxel, two therapeutic agents routinely used for the treatment of ovarian cancer. In order to index the chemoresponse, we analyzed the spatiotemporal evolution of the mortality fraction, as judged by vital dyes and confocal microscopy, within spheroids subjected to different drug concentrations and treatment durations inside the microfluidic device. To reflect microenvironment effects, we tested the effect of exogenous extracellular matrix and serum supplementation during spheroid formation on their chemotherapeutic response. Spheroids displayed augmented chemoresistance in comparison to monolayer culturing. This resistance was further increased by the simultaneous presence of both extracellular matrix and high serum concentration during spheroid formation. Following exposure to chemotherapeutics, cell death profiles were not uniform throughout the spheroid. The highest cell death fraction was found at the center of the spheroid and the lowest at the periphery. Collectively, the results demonstrate the validity of the approach, and provide the basis for further investigation of chemotherapeutic responses in ovarian cancer using microfluidics technology. In the future, such microdevices could provide the framework to assay drug sensitivity in a timeframe suitable for clinical decision making.

Eluding antibiotic resistance: capitalizing on antimicrobial peptides interaction with the lipid bilayer

It is widely accepted that the emergence of drug-resistant pathogens is the result of the overuse and misuse of antibiotics. Infectious Disease Society of America, Center for Disease Control and World Health Organization continue to view, with concern, the lack of antibiotics in development, especially those against Gram-negative bacteria. Antimicrobial peptides (AMPs) have been proposed as an alternative to antibiotics due to their selective activity against microbes and minor ability to induce resistance. For example, the Food and Drug Administration approved Daptomycin (DAP) in 2003 for treatment of severe skin infections caused by susceptible Gram-positive organisms. Currently, there are 12 to 15 examples of modified natural and synthetic AMPs in clinical development. But most of these agents are against Gram-positive bacteria. Therefore, there is unmet medical need for antimicrobials used to treat infections caused by Gram-negative bacteria. In this study, we show that a pro-apoptotic peptide predominantly used in cancer therapy, (KLAKLAK)2, is an effective antimicrobial against Gram-negative laboratory strains and clinical isolates. Despite the therapeutic promise, AMPs development is hindered by their susceptibility to proteolysis. Here, we demonstrate that an all-D enantiomer of (KLAKLAK)2, resistant to proteolysis, retains its activity against Gram-negative pathogens. In addition, we have elucidated the specific site and mechanism of action of D(KLAKLAK)2 through a repertoire of whole-cell and membrane-model assays. Although it is considered that development of resistance does not represent an obstacle for AMPs clinical development, strains with decreased susceptibility to these compounds have been reported. Staphylococci resistance to DAP was observed soon after its approval for use and has been linked to alterations of the cell wall (CW) and cellular membrane (CM) properties. Immediately following staphylococcal resistance, Enterococci resistance to DAP was seen, yet the mechanism of resistance in enterococci remains unknown. Our findings demonstrate that, similar to S. aureus, development of DAP-resistance in a vancomycin-resistant E. faecalis isolate is associated with alterations of the CW and properties of the CM. However, the genes linked to these changes in enterococci appear to be different from those described in S. aureus.

Characterization of the beta-lactamase gene from {\it Enterococcus faecalis\/} strain HH22

The plasmid-encoded, constitutively produced $\beta$-lactamase gene from Enterococcus faecalis strain HH22 was genetically characterized. A restriction endonuclease map of the 5.1 kb EcoRI fragment encoding the enterococcal $\beta$-lactamase was prepared and compared with the restriction map of a cloned staphylococcal $\beta$-lactamase gene (from the naturally-occurring staphylococcal $\beta$-lactamase plasmid pI258). Comparison and hybridization studies showed that there were identical restriction sites in the region of the $\beta$-lactamase structural gene but not in the region surrounding this gene. Also the enterococcal $\beta$-lactamase plasmid did not encode resistance to mercury or cadmium which is encoded by the small, transducible staphylococcal $\beta$-lactamase plasmids. The nucleotide sequence of the enterococcal gene was shown to be identical to the published sequences of three of four staphylococcal type A $\beta$-lactamase genes; more differences were seen with the genes for staphylococcal type C and D enzymes. One hundred-forty nucleotides upstream of the $\beta$-lactamase start codon were also determined for the inducible staphylococcal $\beta$-lactamase gene on pI258; this sequence was identical to that of the constitutively expressed enterococcal gene indicating that the changes resulting in constitutive expression are not due to changes in the promoter or operator region. Moreover, complementation studies indicated that production of the enterococcal enzyme could be repressed. The gene for the enterococcal $\beta$-lactamase and an inducible staphylococcal $\beta$-lactamase were each cloned into a shuttle vector and then transformed into enterococcal and staphylococcal recipients. The major difference between the two host backgrounds was that more enzyme was produced by the staphylococcal host, regardless of the source of the gene but no qualitative difference was seen between the two genera. Also a difference in the level of resistance to ampicillin was seen between the two backgrounds with the cloned enzymes by MIC and time-kill studies. The location of the enzyme was found to be host dependent since each cloned gene generated extracellular (free) enzyme in the staphylococcus and cell bound enzyme in the enterococcus. Based on the identity of the enterococcal $\beta$-lactamase and several staphylococcal $\beta$-lactamases, these data suggest recent spread of $\beta$-lactamase to enterococci and also suggest loss of a functional repressor. ^

On the Pulling Power of Ice Streams

Gravity wants to pull an ice sheet to the center of the Earth, but cannot because the Earth's crust is in the way, so ice is pushed out sideways instead. Or is it? The ice sheet "sees" nothing preventing it from spreading out except air, which is much less massive than ice. Therefore, does not ice rush forward to fill this relative vacuum; does not the relative vacuum suck ice into it, because Nature abhors a vacuum? If so, the ice sheet is not only pulled downward by gravity, it is also pulled outward by the relative vacuum. This pulling outward will be most rapid where the ice sheet encounters least resistance. The least resistance exists along the bed of ice streams, where ice-bed coupling is reduced by a basal water layer, especially if the ice stream becomes afloat and the floating part is relatively unconfined around its perimeter and unpinned to the sea floor. Ice streams are therefore fast currents of ice that develop near the margins of an ice sheet where these conditions exist. Because of these conditions, ice streams pull ice out of ice sheets and have pulling power equal to the longitudinal gravitational pulling force multiplied by the ice-stream velocity. These boundary conditions beneath and beyond ice streams can be quantified by a basal buoyancy factor that provides a life-cycle classification of ice streams into inception, growth, mature, declining and terminal stages, during which ice streams disintegrate the ice sheet. Surface profiles of ice streams are diagnostic of the stage in a life cycle and, hence, of the vitality of the ice sheet.

Mechanisms and evolution of plant resistance to aphids

Aphids are important herbivores of both wild and cultivated plants. Plants rely on unique mechanisms of recognition, signalling and defence to cope with the specialized mode of phloem feeding by aphids. Aspects of the molecular mechanisms underlying aphid-plant interactions are beginning to be understood. Recent advances include the identification of aphid salivary proteins involved in host plant manipulation, and plant receptors involved in aphid recognition. However, a complete picture of aphid-plant interactions requires consideration of the ecological outcome of these mechanisms in nature, and the evolutionary processes that shaped them. Here we identify general patterns of resistance, with a special focus on recognition, phytohormonal signalling, secondary metabolites and induction of plant resistance. We discuss how host specialization can enable aphids to co-opt both the phytohormonal responses and defensive compounds of plants for their own benefit at a local scale. In response, systemically induced resistance in plants is common and often involves targeted responses to specific aphid species or even genotypes. As co-evolutionary adaptation between plants and aphids is ongoing, the stealthy nature of aphid feeding makes both the mechanisms and outcomes of these interactions highly distinct from those of other herbivore-plant interactions. © 2016 Macmillan Publishers Limited.

Retrospective study of prevalence of Community Associated Methicillin Resistant Staphylococcus Aureus (CA-MRSA) seen in Texas MedClinics, San Antonio, TX from April 2006-August 2009

Purpose. To evaluate the presence of Community Associated–Methicillin Resistant Staphylococcus Aureus, CA-MRSA, in abscesses and skin and soft tissue infections presenting at 9 urgent care clinics in San Antonio, TX. ^ Methods. During the 40-month retrospective study (April 2006 to August 2009), wound cultures collected in 9 urgent care centers were evaluated for MRSA growth, antibiotics prescribed, follow up wound care, and antibiotic prescribing habits by physicians for all patients presenting with abscesses and skin/soft tissue infections. ^ Results. Across 9 urgent care centers in San Antonio, TX, 36,797 abscesses and cases of skin and soft tissue infections were treated during 40 months. Of the 36,797 cases, 9290 patients had wound cultures sent with 5,630 cultures sent to Texas MedClinic’s primary lab. Of the 5630 cultures sent to their primary lab, this reflected a prevalence of 4727 (84 %) cultures were positive for MRSA. Of the 9290 patients who had a wound culture sent (April 10th, 2006 to August 31st, 2009), a total of 4,307 antibiotics were prescribed. The top five antibiotics prescribed for CA-MRSA were Bactrim (55.5%), Clindamycin (18.4%), Bactroban (5%), Amoxicillin (3.5%), and Doxycycline (3%) representing 85.4% of the antibiotics prescribed. 8809/9290 (94.8%) of patients required no more than 3 follow up visits. Of the 33 physicians working full time during the entire study period, 29/33 (87.8%) of the physicians were family medicine physicians and represented varied prescribing rates of antibiotics between 11-76% with 26/33 (78.8%) of physicians prescribing antibiotics greater than 40% of the time.^ Conclusions. Abscesses and soft tissue infections are a common presenting complaint to urgent care centers. This study reveals that antibiotic-prescribing practices can be improved with physician education since this high prevalence was not known previously. Also, treating abscesses with limited packing has been shown to be a viable option in this particular circumstance and would be open field for additional clinical research. Due to the high prevalence of CA-MRSA skin and soft tissue infections among patients presenting to urgent care centers presumptive treatment for MRSA is indicated. Increasing levels of resistance to penicillin antibiotics is concerning and warrants alternative antibiotic management strategies.^