Use of the PCR-DGGE method for the analysis of the bacterial community structure in soil treated with the cephalosporin antibiotic cefuroxime and/or inoculated with a multidrug-resistant Pseudomonas putida strain MC1

The widespread use of cefuroxime (XM) has resulted in the increase in its concentration in hospital and domestic wastewaters. Due to the limited removal of antibiotics and antibiotic-resistant genes in conventional systems, the drugs enter the surface water and soils. Moreover, the introduction of XM and/or XM-resistant bacteria into soil may cause a significant modification of the biodiversity of soil bacterial communities. Therefore, the goal of this research was to assess the genetic diversity of a bacterial community in the cefuroxime (XM1-1 mg/kg and XM10-10 mg/kg) and/or antibiotic-resistant Pseudomonas putida strain MC1 (Ps - 1.6 × 107 cells/g)-treated soils as determined by the DGGE (denaturing gradient gel electrophoresis) method. The obtained data were also evaluated using a multivariate analysis and the resistance (RS)/resilience (RL) concept. Strain MC1 was isolated from raw sewage in the presence of XM and was resistant not only to this antibiotic but also to vancomycin, clindamycin and erythromycin. The DGGE patterns revealed that the XM10 and XM10+Ps treatments modified the composition of the bacterial community by the alteration of the DGGE profiles as well as a decline in the DGGE indices, in particular on days 30, 60, and 90. In turn, the XM1 and XM1+Ps or Ps treatments did not affect the values of richness and diversity of the soil bacteria members. A principal component analysis (PCA) also indicated that XM markedly changed the diversity of bacterial assemblages in the second part of the experiment. Moreover, there were differences in the RS/RL of the DGGE indices to the disturbances caused by XM and/or Ps. Considering the mean values of the RS index, the resistance was categorized in the following order: diversity (0.997) > evenness (0.993) > richness (0.970). The soil RL index was found to be negative, thus reflecting the progressing detrimental impact of XM on the genetic biodiversity of bacteria within the experiment. These results indicate that the introduction of XM at higher dosages into the soil environment may exert a potential risk for functioning of microorganis

Chironomidae-inferred anthropogenic and natural processes in moat and palaeochannel systems − Rozprza case study

Celem pracy jest paleoekologiczna rekonstrukcja środowiska w rejonie stanowiska archeologicznego w Rozprzy (woj. łódzkie), na podstawie analizy subfosylnych zgrupowań Chironomidae (Diptera). Zmienność zgrupowań Chironomidae w późnym glacjale zbadano z wykorzystaniem profilu jeziornego pobranego z paleomeandru rzeki Luciąży. Odtworzono średnie temperatury lata i zmiany hydrologiczne w okresie ok. 13 300 – 12 200 lat cal. BP, wraz z jednym z pierwszych dobrze udokumentowanych stwierdzeń chłodnej oscylacji interallerødzkiej w Europie Środkowo-Wschodniej. Stosunkowo wysokie temperatury lata (do 16˚C) w młodszym dryasie mogą wskazywać na lokalnie cieplejsze warunki pod koniec zlodowacenia wisły bądź wysoki stopień kontynentalizmu w tym stadiale w Polsce środkowej. Wyniki analiz Chironomidae umożliwiły rekonstrukcję zmian siedliskowych w paleostarorzeczu i fosie, wchodzącej w skład kompleksu późnośredniowiecznego gródka. W zbiorniku sztucznym (fosa) wykazano znacznie szybsze tempo paludyfikacji niż w starorzeczu. W naturalnych zbiornikach zgrupowania Chironomidae były kształtowane głównie przez globalne i regionalne zmiany klimatyczne. W fosie determinantami występowania i składu Chironomidae były lokalne zmiany siedliskowe, takie jak zmiany trofii wody oraz przepłukiwanie fosy. Analiza osadów organicznych z czterech rdzeni pobranych w obrębie systemu fos, pozwolił na scharakteryzowanie zmian siedliskowych w czasie i przestrzeni. Nawet w tak małym i płytkim zbiorniku, tempo i zakres procesów ekologicznych nie były równomierne. Zróżnicowanie fosy zależało głównie od głębokości dna, ekspozycji na procesy denudacyjne i zaopatrzenia w świeżą wodę. W pracy przedstawiono również wyniki analizy grup morfologicznych i troficznych Chironomidae, będącej pierwszym etapem badań nad wykorzystaniem ich cech funkcyjnych w paleoekologii. Udowodniono, że morfo-taksonomia może być wykorzystywana do rekonstrukcji zmian siedliskowych w podobnym stopniu, jak gildie troficzne.This work was supported by National Science Centre grant [DEC-2013/11/B/HS3/03785] and Faculty of Biology and Environmental Protection, University of Lodz subsidies of for young scientists [B1711000001557.02, B1811000001847.02, B1911000002143.02

An analysis of the effects of vancomycin and/or vancomycin-resistant Citrobacter freundii exposure on the microbial community structure in soil

The occurrence of antibiotics and antibiotic resistance genes in the environment has become a subject of growing concern. The extensive use of vancomycin and other pharmaceuticals may alter the biodiversity of soil microbial communities and select antibiotic-resistant bacteria. Therefore, the purpose of the study was to evaluate the impact of vancomycin and/or vancomycin-resistant Citrobacter freundii on soil microbial communities using the denaturing gradient gel electrophoresis (DGGE) and the phospholipid fatty acid (PLFA) approaches. The experiment had a completely randomized block design with the following treatments: Control soil (C), soil with vancomycin (1 mg/kg soil-VA1), soil with vancomycin (10 mg/kg soil-VA10), soil with C. freundii (Cit), soil with vancomycin (1 mg/kg soil) and C. freundii (VA1+Cit), and soil with vancomycin (10 mg/kg soil) and C. freundii (VA10+Cit). A bacterial strain resistant to vancomycin was isolated from raw sewage collected from the municipal sewage treatment plant. The obtained results indicated that the antibiotic and/or the bacterial strain exerted a selective pressure that resulted in qualitative and quantitative changes in the population of soil microorganisms. However, a multivariate analysis showed that the genetic and structural diversity of the soil microbial community was primarily affected by the incubation time and to a lesser extent by the antibiotic and introduced bacteria. DGGE analysis clearly showed that certain species within the bacterial community were sensitive to vancomycin as was evidenced by a decrease in the values of S (richness) and H (Shannon-Wiener) indices. Moreover, a PLFA method-based analysis revealed alterations in the structure of the soil microbial community as indicated by changes in the biomass of the PLFA biomarkers specific for Gram-positive and Gram-negative bacteria as well as fungi. The changes observed in the community of soil microorganisms may decrease the rate of microbial-mediated processes, which can lead to a disturbance in the ecological balance of the soil ecosystem

Functional Diversity of Soil Microbial Communities in Response to the Application of Cefuroxime and/or Antibiotic-Resistant Pseudomonas putida Strain MC1

Cefuroxime (XM), the most commonly prescribed antibiotic from the cephalosporin group, may cause changes in the structure of the soil microbial community, and these changes may also be reflected in the alteration of its functionality. Therefore, due to the lack of studies on this topic, the scope of this study was to assess the functional diversity and catabolic activity of the microbial community in soil treated with XM (1 mg/kg and 10 mg/kg soil) using the community-level physiological profile (CLPP) approach during a 90-day experiment. In addition, the effect of antibiotic-resistant Pseudomonas putida strain MC1 (Ps) was also evaluated. The resistance/resilience concept and multifactorial analysis were used to interpret the data. The results showed that the introduction of XM and/or Ps into the soil caused changes in the catabolic activity and functional diversity of the microbial community. A decrease in the values of the CLPP indices (i.e., microbial activity expressed as the average well-color development (AWCD), substrate richness (R), the Shannon-Wiener (H) and evenness (E) indices and the AWCD values for the six carbon substrate groups) for the XM-treated soil was generally detected up to 30 days. In turn, at the same time, the activity measured in the Ps-inoculated soil was higher compared to the control soil. A stimulatory effect of XM at 10 mg/kg (XM10) and XM10+Ps on the utilization pattern of each substrate group was found at the following sampling times (days 60 and 90). The AWCD values for the utilization of amines, amino acids, carbohydrates, carboxylic acids, miscellaneous compounds and polymers for these treatments were found to be up to 2.3-, 3.1-, 2.3-, 13-, 3.4- and 3.3-fold higher compared to the values for the nontreated control, respectively. The resistance of the CLPP indices and the AWCD values for the carbon substrate groups were categorized as follows: E > H > R > AWCD and amino acids = carbohydrates > polymers > amines > miscellaneous > carboxylic acids, respectively. The results suggest a low initial resistance of the soil microbial community to XM and/or Ps, and despite the short-term negative effect, the balance of the soil ecosystem may be disturbed

Dawkowanie i koszt lanreotydu Autogel 120 mg stosowanego w leczeniu akromegalii w Polsce — wyniki 2-letniego badania Lanro-Study

Introduction: To evaluate, over 24 months of prospective follow-up, the dosage and costs of lanreotide AUTOGEL 120 mg (ATG120) administered as part of routine acromegaly care in Poland.Material and methods: A multicentre, non-interventional, observational prospective study on resource utilisation in Polish acromegalic patients treated with ATG120 at 4-week or extended (> 4 weeks) dosing interval. The study population consisted of adult acromegalic patients treated for at least 3 injections of ATG120. The endpoints were: percentage of patients treated with ATG120 at an extended dosing interval (> 4 weeks), mean time between injections, and the cost of ATG120 during a 24-month prospective observation. Costs were calculated in PLN from the public health-care payer and patient perspective for the year 2014.Results: 143 patients were enrolled in, and 132 completed (70% women, 81% macroadenoma, 75% previous surgery) the analysis. During two years, changes in the treatment pattern were reported in 41 patients: 17% of them had increased injection interval and 10% switched to octreotide LAR and then returned to ATG120. Sixty-three patients (48%) received ATG120 at an extended dosing interval. ATG120 was predominantly administered in an out-patient setting (84%) by a health care professional (97%).Conclusions: The results demonstrated that extended dosing interval of ATG120 is used in a substantial proportion of patients in routine clinical practice in Poland. Such findings support the potential for ATG120 in reducing treatment burden in the real-world clinical environment. (Endokrynol Pol 2015; 66 (2): 142–148)Wstęp: Ocena schematu dawkowania i kosztu lanreotydu Autogel 120 mg (ATG120) stosowanego w leczeniu akromegalii w Polsce.Materiał i metody: Wieloośrodkowe, nieinterwencyjne, obserwacyjne badanie prospektywne oceniające zużycie zasobów ochrony zdrowia w populacji polskich pacjentów z akromegalią leczonych ATG120 z zastosowaniem 4-tygodniowego lub wydłużonego (> 4 tygodni) odstępu między dawkami leku. Do badania włączani byli dorośli pacjenci, którzy otrzymali co najmniej 3 dawki ATG120. Horyzont czasowy badania wynosił 24 miesięcy. Koszt leku obliczono z perspektywy płatnika publicznego i pacjenta w 2014 roku.Wyniki: Do badania włączono 143 pacjentów, 132 z nich ukończyło badanie (70% kobiet, u 81% makrogruczolak, 75% było wcześniej leczonych chirurgicznie). W czasie 2 lat zmiany schematu leczenia dokonano u 41 chorych, u 17% z nich wydłużono odstęp między dawkami, u 10% — zmieniono lek na oktreotyd LAR a potem ponownie zastosowano ATG120. U 63 pacjentów (48%) odstęp pomiędzy dawkami ATG120 wynosił > 4 tygodnie. ATG120 był przeważnie podawany ambulatoryjnie (84%) przez fachowy personel medyczny (97%). Wnioski: Badanie wykazało, że w codziennej praktyce klinicznej u znaczącego odsetka pacjentów odstęp czasowy pomiędzy kolejnymi dawkami ATG120 jest dłuższy niż 4 tygodnie. Wynik ten potwierdza tezę, że dzięki stosowaniu ATG120 można obniżyć koszty leczenia akromegalii. (Endokrynol Pol 2015; 66 (2): 142–148