Václav Jedlička

The anatomical relationship between the saphenous nerve and the great saphenous vein

Objective Damage to the saphenous nerve (SN) has been a known complication during varicose vein surgeries. We tested whether a better knowledge of the anatomy of the SN and the great saphenous vein (GSV) can prevent such damage. Methods We conducted a morphological and histological examination on 86 limbs from 43 cadavers in order to analyse the anatomical interrelation between the SN and the GSV in the lower leg and we also measured the distance between the nerve and the vein in a sample of 42 sections from three parts of the lower leg. Results The anatomical relationship between the SN and the GSV is varied and the two structures run close to each other so a better knowledge of their anatomy in itself proved insufficient in preventing damage to the SN. Conclusion However, in the case of endovenous laser therapy and radiofrequency ablation tumescent anaesthesia decreases the risk of damage to the SN.

Comparison of Two Non-Invasive Methods of Microbial Analysis in Surgery Practice: Incision Swabbing and the Indirect Imprint Technique

BACKGROUND A variety of methods exist to take samples from surgical site infections for cultivation; however, an unambiguous and suitable method has not yet been defined. The aim of our retrospective non-randomized study was to compare two non-invasive techniques of sampling material for microbiologic analysis in surgical practice. We compared bacteria cultured from samples obtained with the use of the swab technique, defined in our study as the gold standard, with the indirect imprint technique. METHODS A cotton-tipped swab (Copan, Brescia, Italy) was used; the imprints were taken using Whatman no. 4 filter paper (Macherey-Nagal, Duren, Germany) cut into 5×5 cm pieces placed on blood agar in a Petri dish. To culture the microorganisms in the microbiology laboratory, we used blood agar, UriSelect 4 medium (Bio-Rad, Marnes-la-Coquette, France), and a medium with sodium chloride (blood agar with salt). After careful debridement, a sample was taken from the incision surface by swab and subsequently the same area of the surface was imprinted onto filter paper. The samples were analyzed in the microbiology laboratory under standard safety precautions. The cultivation results of the two techniques were processed statistically using contingency tables and the McNemar test. Those samples that were simultaneously cultivation-positive by imprint and -negative by swabbing were processed in greater detail. RESULTS Over the period between October 2008 and March 2013, 177 samples from 70 patients were analyzed. Sampling was carried out from 42 males and 28 females. One hundred forty-six samples were from incisions after operations (21 samples from six patients after operation on the thoracic cavity, 73 samples from 35 patients after operation on the abdominal cavity combined with the gastrointestinal tract, 52 samples from 19 patients with other surgical site infections not included above) and 31 samples from 11 patients with no post-operative infection. One patient had a sample taken both from a post-operative and a non-post-operative site. Coincidently, the most frequent cultivation finding with both techniques was a sterile one (imprint, 62; swab, 50). The microorganism cultivated most frequently after swabbing was Pseudomonas aeruginosa (22 cases), compared with Escherichia coli when the filter paper (imprint) was used (31 cases). The imprint technique was evaluated as more sensitive compared with swabbing (p=0.0001). The κ statistic used to evaluate the concordance between the two techniques was 0.302. Of the 177 samples there were 53 samples simultaneously sterile using the swab and positive in the imprint. In three samples colony- forming units (CFU) were not counted; 22 samples were within the limit of 0-25×10(1) CFU/cm(2), 20 samples within the limit of 25×10(1)-25×10(2) CFU/cm(2), five within the limit of 25×10(2)-25×10(3) CFU/cm(2), and three of more than 25×10(4) CFU/cm(2). CONCLUSIONS The hypothesis of swabbing as a more precise technique was not confirmed. In our study the imprint technique was more sensitive than swabbing; the strength of agreement was fair. We obtained information not only on the type of the microorganism cultured, but also on the number of viable colonies, expressed in CFU/cm(2).

Indirect imprint sampling technique using a filter paper pad for microbial analysis

Compounds present in oil sludge such as polycyclic aromatic hydrocarbons (PAHs) are known to be cytotoxic, mutagenic and potentially carcinogenic. Microorganisms including bacteria and fungi have been reported to degrade oil sludge components to innocuous compounds such as carbon dioxide, water and salts. In the present study, we isolated different bacteria with PAH-degrading capabilities from compost prepared from oil sludge and animal manures. These bacteria were isolated on a mineral base medium and mineral salt agar plates. A total of 31 morphologically distinct isolates were carefully selected from 5 different compost treatments for identifi cation using polymerase chain reaction (PCR) of the 16S rRNA gene with specifi c primers (universal forward 16S-P1 PCR and reverse 16S-P2 PCR). The amplicons were sequenced and sequences were compared with the known nucleotides from the GenBank. The phylogenetic analyses of the isolates showed that they belong to 3 different clades; Firmicutes, Proteobacteria and Actinobacteria. These bacteria identifi ed were closely related to the genera Bacillus, Arthrobacter, Staphylococcus, Brevibacterium, Variovorax, Paenibacillus, Ralstonia and Geobacillus. The results showed that Bacillus species were predominant in all composts. Based on the results of the degradation of the PAHs in the composts and results of previous studies on bacterial degradation of hydrocarbons in oil, the characteristics of these bacterial isolates suggests that they may be responsible for the breakdown of PAHs of different molecular weights in the composts. Thus, they may be potentially useful for bioremediation of oil sludge during compost bioremediation. Unauthenticated Download Date | 10/19/16 10:12 PM 68 O. Ubani, H.I. Atagana, M.S. Thantsha and thermophilic activities. Microbial activities in compost generate high temperature, which increases solubility of contaminants and induces microbial co-metabolic activity (Sheetal 2012). In compost, there is abundance of nutrients and organic matter that have effect on microbial degradation of oil sludge constituents (Sheetal 2012). As the compost matures, the pollutants are degraded, digested, metabolised and transformed into humus as well as inert products such as carbon dioxide, water and salts. In this study, the initial concentrations of the PAHs (2 to 6 rings) detected were between 1.44 mg/kg to 205.81 mg/kg before the co-composting process of oil sludge. The results obtained showed reduction in selected PAHs (66% to 80%) in all co-composting piles over a period of ten months, as shown in Table 3. This result is in agreement with the report from comparison of bioaugmentation and composting for remediation of oil sludge (Ouyang et al. 2005). The results obtained from the compost piles showed that composting can be used to degrade PAHs present in oil sludge, suggesting the existence of active bacterial community in the compost (Katsivela et al. 2003). This study was to determine whether microbial growth and activities were enhanced as well as bacteria community involved in the degradation of oil sludge (PAHs) during the composting period. Culture dependent and non-culture dependent methods have been used to determine the composition of the metabolically active microorganisms that played an important role in the degradation of pollutants (Saman 2010). Culture dependent methods (CDM) often involve proper isolation and in vitro cultivation of microorganisms based on their required growth factors (Saman 2010), while non-culture dependent method is used for identifi cation of uncultivable microorganism. These methods complement each other to enable the identifi cation of more than 90% microorganisms. Perhaps, the most widely used non-culture dependent method is the molecular method. This usually involves direct DNA extraction from samples, PCR amplifi cation and nucleotide sequence for identifi cation of microbial diversity based on the analysis of the 16SrRNA gene sequences (Mokno-Tlili et al. 2009, Inceoglu et al. 2010). It is a reliable method that can be used to detect diversity, quantity and sometimes viability of microbial contents of a sample (Inceoglu et al. 2010, Saman, 2010). However, it seems that combination of both methods is always essential in isolation and characterisation of bacterial isolates. The aim of this study was to isolate, identify and characterise bacterial consortium present in a compost system with potentials to degrade PAHs present in oil sludge. Materials and methods Soil Garden soil was air-dried and analysed to determine the soil type, organic carbon content, total nitrogen content, total phosphorus content (C; N and P), soil pH and water holding capacity (WHC), using concentrated acid digestion method (CADM) with Induction Coupled Plasma (ICP-OES Optima 4300 DV, Perkin Elmer, Waltham, MA, USA). Oil sludge Oil sludge from a refi nery was soxhlet extracted with dichloromethane as the solvent and characterized by gas chromatography/mass spectrometry (GC/MS) method (US EPA 827Asymptomatic urinary tract infections resulting from poor diagnosis in pregnant women poses a serious threat of complications during pregnancy. This study reports the prevalence of Urinary Tract Infections (UTIs) and the bacteriological causative agents among pregnant women attending antenatal clinics at a Tertiary Care Hospital in Ile Ife, South western Nigeria. 335 mid-stream clean catch urine samples were collected from pregnant women and cultured for the presence of bacterial pathogens. All the isolates were subjected to antibiotic sensitivity testing on Mueller Hinton agar by modified Kirby-bauer disk diffusion technique using CLSI, guidelines. Data were analyzed using SPSS for Windows version 16. A total of 190 showed significant bacterial growth while 145 showed no significant bacterial growth. Bacterial agents isolated included Escherichia coli showing the highest occurrence of [60(31.5%) followed by Klebsiella spp and Staphylococcus aureus with prevalences of [49(25.7%) and [32(16.8%) respectively. Other organisms implicated are Proteus mirabilis [20(10.5%), Coagulase Negative Staphylococci [24(12.3%) and Pseudomonas aeruginosa with the least prevalence of [5(2.6%). Resistant to antibiotics such as amoxycilin 64%, gentamicin 52.6%, erythromycin 62.5%, ceftazidime 69%, cefotaxime 74%, ceftriaxone 79.6% while a high sensitivity to tetracycline (88.5%), nitrofurantoin (96.3%), imipenem (100%) respectively. This study indicated a high prevalence of UTIs (56.7%) in pregnant women though most of them showed no clinical manifestation. Therefore, urine microbial screening should be included in the routine antenatal checkups for pregnant women to detect the asymptomatic infections to reduce its risk to pregnancies.Introduction There exists a variety of methods how to take samples from surgical wounds for cultivation; nevertheless, an unambiguous and most suitable procedure has so far been not defined with satisfactory precision. The aim of this presentation is to introduce an indirect imprint technique using filter paper pad for microbial analysis of wounds (shows principles of collecting, storaging, transporting, evaluation of samples and interpreting obtained information with all its advantages and disadvantages) and compare it with routinely used swab Method Imprints were carried out using the filter paper Whatman No. 4, cut to a size of 5 x 5 cm and previously sterilized in an autoclaved at 120 oC for 20 minutes and aseptically attached to 24 hour-old blood agar in a Petri dish, for the swabs, we used sterile cotton tips made by the Italian company COPAN; After careful debridement, a sample was taken from the wound surface by the swab and subsequently the same area of the investigation surface was imprinted onto a filter paper. Sheet of filter paper was taken from Petri dish with blood agar by sterile tweezers and put on to surface of wound. After 10 seconds paper became wet and was moved back to the Petri dish and then transported to a microbial laboratory as soon as possible. In the microbial laboratory a filter paper was removed from Petri dish and put on to new blood agar for 10 seconds and then to UriSelect 4 medium and after then to NaCl (salt) blood agar. The cultivation was done in the atmosphere with 10 % CO2 and temperature 37 oC. The final control of Petri dishes happened 48 hours later. The cultivation results of the two techniques were statistically processed using contingency tables and McNemar test. Those samples that were simultaneously cultivation positive in the imprint and negative in the swab were processed in greater detail. Results Result of an indirect imprint was semi-quantitative, one part of the result was type of microbe cultivated from wound and other was density of them accounted by colony forming unit (CFU), an estimate of viable bacterial numbers in wound, the results were given as CFU/cm2. 177 samples from 70 patients were analyzed. Sampling was carried out in 42 men and 28 women. The identical results of both culture methods were in 114 samples and different in 63 samples. The most frequent result of swab was sterile result, in 62 cases; imprint was sterile in 50 cases. Imprint and swab were both negative in 46 cases, and positive in 68 cases. In 15 cases there was a negative imprint and positive swab and in 48 cases there was positive imprint and negative swab. Coincidently, the most frequent cultivation finding with both techniques was a sterile one. The most frequently cultivated microorganism after using swab was Pseudomonas aeruginosa (in 22 cases), compared with Escherichia coli when the filter paper (imprint) was used (in 31 cases). The imprint technique was evaluated as more sensitive compared to swab, at the level of statistical significance P = 0.0001. Of the 177 samples there were 53 samples sterile in the swab and positive in the imprint simultaneously. In three samples CFU was not counted, 22 samples were within the limit of 0 - 25 x 101 CFU/cm2, 20 samples within the limit of 25 x 101 – 25 x 102 CFU/cm2, five within the limit of 25 x 102 – 25 x103 CFU/cm2, and three of more than 25 x 104 CFU/cm2. Conclusion In our study the imprint technique appears as a highly more sensitive than swabbing. We obtain information not only on the type of the microorganism cultured, but also on the number of viable colonies per square unit, expressed in CFU/cm2.Malaria and anaemia during pregnancy is still a major health problem in endemic countries with clinical consequences including death of both mother and child. In Nigeria, statistics shows that as many as 300,000 lives especially those of children and pregnant women are lost annually due to malaria. This study was aimed at assessing the impact of malaria and anaemia among pregnant women living in Calabar South Local Government Area of Cross River State, Nigeria, which is characterized by unstable transmission of malaria. .A total of 664 subjects were enrolled in the study made up of 414 pregnant women attending antenatal clinic in the University of Calabar Teaching Hospital Calabar, Nigeria and 250 age-matched non-pregnant women served as control group. Full blood count was done using PCE-210 automatic cell counter, malaria parasite detection was through examination of peripheral blood smears and malaria parasite count/density was done using WHO standard method (WHO, 1991). Anaemia was significantly(P <0.05) higher among the pregnant women 253(61.1)(Hb<11g/dl) than in the non-pregnant women 96(38.3%)(Hb<12g/dl).The prevalence of malaria parasite infection was 290(70.1%) in pregnant women and 152(60.8%) in the control group. Prevalence of anaemia and malaria parasite was found to be higher in the primigravidae than in the multigravidae. Primigravidae were more susceptible to the parasite especially Plasmodium falciparum with mean parasite density of 1962.50 ± 220.90 (parasite/µl) than the multigravidas with parasite density 446.70 ± 296.90 (parasite/µl). Malaria parasite density increased significantly with gestational age but anaemia was more prevalent in the second trimester than in the other trimesters. There was a negative correlation between haemglobin and malaria parsite density in both pregnant and non-pregnant women (r = -0.1964). The results showed that malaria infection caused by P. falciparum had serious effect on pregnant women living in the study area. Malaria in pregnancy should be recognized as a global priority in health care services.The study advocate the need for pregnant women to undergo routine haemoglobin estimation and early malaria prophylaxis considering the deleterious effects of anaemia on them and their foetus.A identification of coagulase-negative staphylococci (CoNS) is important especially when they are isolated from multiple blood cultures of a patient. Maldi-Tof MS is a new fast and accurate technique which allows examination of protein profiles of bacteria. This study aimed to compare two methods; Maldi-Tof MS (Bruker Daltonics) and BD Phoenix (Becton Dickinson) for identification of CoNS isolates from blood culture. Totally 102 CoNS bloodstream isolates representing 9 species were analyzed in this study. There was 73.5% correlation between two methods. The 10 S.hominis described by Maldi-Tof MS were identified as 3 S.epidermidis, 5 S.saprophyticus, 1 S.capitis, and 1 S.haemolyticus; 12 S.epidermidis identified as 3 S.saprophyticus, 2 S.haemolyticus 1 S.capitis, 1 S.hominis, 1 S.chromotogenes, 1 S.warneri, 1 S.hyicus, 1 S.aureus, and 1 S.capitis; 4 S.haemolyticus were identified as 2 S.epidermidis, and 2 S.hominis; 1 S.warneri was identified as S.capitis by BD Phoenix system. According to the previous studies Maldi-tof MS seemed to be a powerful method in ID of CoNS with up to 99.3% correct results. The next step of this study will be tuf sequencing to find the accuracy rates of these two methods. Mustafa Guney, J Microb Biochem Technol 2013, 5:4 http://dx.doi.org/10.4172/1948-5948.S1.010V species are the leading cause of bacterial illness (V. parahaemolyticus) and mortality (V. vulnificus) associated with seafood consumptionin the United States. Pathogenicity of these organisms is not fully understood, likely due to the involvement of multiple factors. Some virulence determinants have been identified for these organisms; however, no one virulence determinant, or set of determinants, has been shown to be required to cause illness. Additionally, isolates from patients with disease can lack any known virulence markers. In other instances, areas of the country with the greatest incidence of illness have lower levels of strains with identified virulence determinants in environmental samples. Taken together, these data highlight the necessity for identification of more robust virulence determinants in V. parahaemolyticus and V. vulnificus. Our laboratory has undertaken the quest for identification of new virulence biomarkers in these two pathogens using well characterized sets of clinical (presumed virulent) and environmental (presumed mostly avirulent) isolates. Using these strain panels, multiple approaches are being used to identify distinguishing features of the virulent (clinical) strains. Methods currently being employed include molecular subtyping, proteomics, and whole genome sequencing. A brief history of the understanding of vibrio virulence will be presented, followed by the detailed approach for identification of new biomarkers undertaken in our laboratory and resultant findings. Jessica L. Jones, J Microb Biochem Technol 2013, 5:4 http://dx.doi.org/10.4172/1948-5948.S1.010

182P LARGE CHEST WALL RESECTION FOR THE PRIMARY CHEST WALL NEOPLASM – TECHNIQUE AND RESULTS

Sarkomy hrudni stěny jsou raritni. Jsou vsak velmi zavažne, život ohrožujici. Odpověď nadorů na chmeoterapii i radioterapii neni uspokojiva.Resekce hrudni stěny s rekonstrukci za použiti synteticke siťky a svaloveho laloku představuje bezpecnou a efektivni terapii. I rozsahle resekce nejsou zatiženy zvýsenou peropacni morbiditou ci mortalitou.

140P THE ROLE OF THORACIC SURGERY IN TREATMENT OF PULMONARY METASTASIS

Background The problem of surgical treatment of pulmonary metastasis is still alive, it is accepted in the world as a curative way of treatment of pulmonary metastasis. But the problem lies in the Czech Republic that only a few oncologists accept it as one of the most effective ways of treatment. Prognostic factors for pulmonary metastasectomies are complete resection, disease free interval, the amount and size of the metastasis, tumor double time, histology of the primary tumor and biologic aggressiveness of the tumor. Methods The effect of the surgery is based on exact diagnostic investigation (X-ray, HRCT, PET-CT) and on the proper selection of patients based on the contol of the primary tumor, no metastasis in other localizations, the possibility of complete resection, good cardiopulmonal condition of the patient, no other better way of treatment and low-operative risk. As contraindications are mostly considered low pulmonary function, impossibility of complete resection and the presence of many tiny metastasis. According to the size, number and side is chosen the right access to the pulmonary cavity and the proper surgery is performed. The importance of mediastinal lymphadenectomy was not defined yet. The presence of positive lymphnodes is not a good prognostic factor for long-time survival rate. Results The authors will present the results of pulmonary metastasectomies in the last ten years on over 100 patients performed in our department. Most of the surgeries were performed by atypic resection with safety margin, also some lobectomies and enclueations. No pneumonectomy was performed. Mediastinal lymphadenectomy was performed at carcinomas. Conclusions Pulmonary metastasectomy can be when performed by a experienced surgeon and after an exact indication, a way of secure, effective and potentially curative treatment .