En säker bakteriediagnostik kräver en korrekt provtagningsteknik, ett lämpligt transportsätt och rätt odlingsteknik. På grund av centralisering av verksamheter och ekonomiska begräsningar, är transport av prover till mikrobiologilaboratorium mycket vanligt. Detta medför behov av ett transportmedium som håller känsliga bakterier vid liv under långa transport- och förvaringstider. Dessutom skall transportrören klara av maskinell utodling som kräver ett vätskebaserat provtagningsmedium. Denna övergång minskar även arbetsbelastning och förbättra ergonomin hos personalen. I denna studie utvärderades viabilitet av Propionibacterium acnes (n=3), Bacteroides fragilis (n=2) och Neisseria gonorrhoeae (n=3) i transportmediet från COPAN E-swab™. Alla tre bakterier förvarades i transportrören i 24, 48 och 72 timmar (h) och dessutom i 120h för P. acnes och B. fragilis. Den ursprungliga koncentrationen samt den procentuella viabilitetsförlusten över tid beräknades med hjälp av viable count. Efter 120h förvaring i transportmediet hade viabiliteten minskat till 47 – 80 % för P. acnes, 18 % för B. fragilis 1 och 73 % för B. fragilis 2. N. gonorrhoeae visade en minskad viabilitet med 96 – 99,97 % efter 24h förvaring i 4°C och rumstemperatur. P. acnes och B. fragilis kunde förvaras i transportmediet i upp till 5 dygn utan att diagnostiken äventyrades. N. gonorrhoeae kunde förvaras i transportmediet i 24h för att diagnostik genom odling ska vara möjlig. / Safe bacterial diagnostics requires proper sampling techniques, suitable specimen transport and correct inoculation techniques. As a result of centralization of operations and economic restraints, transportation of samples to microbiology laboratories is very common. This entails the need for a transport medium that keeps sensitive bacteria alive under the long transport and storage time. Additionally, the transport tube should be convenient for automatic inoculation that requires a liquid based medium. This transition also reduces workload and improves ergonomics of the staff. In this study the survival of Propionibacterium acnes (n = 3), Bacteroides fragilis (n = 2) and Neisseria gonorrheae (n = 3) in the transport medium Copan E-swab™   was evaluated. All the species were inoculated in the tubes for 24, 48, 72h and also 120h for B. fragilis and P. acnes.  Initial concentration at start time and the percentage survival over time was calculated with the help of viable count. Viability decreased to 47 – 80 % for P. acnes, 18 % for B. fragilis 1 and 73% for B. fragilis 2 after 120 h storage in the transportmedium. N. gonorrhoeae showed a reduction in viability of 96 – 99,97 % after 24h storage in the medium at 4°C and in room temperature. P. acnes and B. fragilis can be stored in the medium for up to 5 days, without endangering a diagnosis. N. gonorrhoeae can be stored in the transport medium for up to 24h to secure a diagnosis through culturing.

Vårdrelaterade infektioner (VRI) är ett ökande problem inom hälso- och sjukvården. På neonatalavdelningen på Länssjukhuset Ryhov i Jönköping har det förekommit inkonsekventa odlingsresultat vid misstänkt VRI orsakad av Staphylococcus aureus. Att förebygga VRI samt värna om patientsäkerheten kräver känsliga och pålitliga laboratorieanalyser. Syftet med studien var att bestämma och jämföra lägsta detektionsintervall för metoderna odling och qPCR med och utan anrikning i MAMSA-buljong, av S. aureus. Seriespädningar av S. aureus tillreddes och koncentrationer för ursprungsrören uträknades via viable count (VC). Odling på blodagar samt qPCR med och utan anrikning i MAMSA-buljong utfördes. Resultaten användes för att bestämma ett lägsta detektionsintervall. Odling gav ett lägsta detektionsintervall mellan 0,5–62 CFU/ml, och qPCR mellan 6400–140 000 CFU/ml. Anrikning med MAMSA-buljong innan qPCR-analys gav ett lägsta detektionsintervall mellan 0,6–140 CFU/ml. För detektion via odling räcker enstaka till tiotals CFU/ml i analysprovet. Vid qPCR behövs tusentals till hundratusentals CFU/ml i analysprovet, men vid qPCR med MAMSA-anrikning kan detektionsintervallet sänkas till nivåer jämförbara med odling. För att kunna applicera studiens resultat på den laborativa verksamheten krävs vidare studier med fler bakteriearter och replikat. / Hospital-acquired infections (HAI) are an increasing problem in health care facilities. In the neonatal intensive care unit at Länssjukhuset Ryhov in Jönköping, inconsistent cultivation results have been observed for suspected HAI caused by Staphylococcus aureus. To prevent HAI and to maintain patient safety, sensitive and reliable laboratory tests are essential. The aim of the study was to determine and compare the lowest range of detection for cultivation and qPCR with and without enrichment in MAMSA broth, for S. aureus. Serial dilutions of S. aureus were made, and the original concentration was determined through viable count. Cultivation on blood agar and qPCR with and without enrichment in MAMSA broth was performed. The results were used to determine a minimum detection range. For cultivation, a minimum detection range of 0,5–62 CFU/ml was determined and for qPCR a range of 6400–140 000 CFU/ml. For qPCR after enrichment in MAMSA broth, a lowest detection range of 0,6–140 CFU/ml was determined. For detection with cultivation single to tenths of CFU/ml were needed in the sample. For qPCR thousands to tenths of thousands of CFU/ml were needed. For qPCR with enrichment in MAMSA broth the detection range could be lowered to levels comparable with cultivation. For clinical application, further studies are needed with more bacterial species and replicates.

Syftet med denna studie var att undersöka den antibakteriella effekten hos ett polyestertyg med tunna invävda koppartrådar. Frågeställningen som skulle besvaras var ifall koppartyget hade en baktericid eller bakteristatisk effekt. Koppartyget är ännu i prototypstadie och om det visar sig ha antibakteriella egenskaper är det tänkt att användas inom klinisk verksamhet för att förhindra bakterietillväxt i sår och andra känsliga lokaler. Koppar är ett essentiellt spårämne, men har också antimikrobiella egenskaper som utövas genom ett brett spektra av mekanismer där skador på cellmembranet är en av de viktigare. Metoderna som användes för att inokulera bakterier på tyget var absorptionsmetoden, där en näringsbuljong innehållande Staphylococcus aureus ATCC 6538 pipetterades på tygproverna, och transfermetoden där tygproverna trycktes mot en agar som racklats med peptonsaltlösning innehållande S. aureus. Totalantalet viabla bakterier per tygprov beräknades efter kort kontakt (<1min) och inkubering (18-24 h vid 37±2°C) genom att räkna viable count. Resultaten efter inkubering visade signifikant skillnad i totalantal bakterier mellan koppartyget och negativ kontroll i tre av fyra försök. Kort kontakt visade tendens till viss antibakteriell effekt. Slutsatsen är att koppartyget skadade och dödade bakterier då de fick inkubera på tyget, medan fler försök behövs för att säkerställa effekten av kort kontakt med koppartyget. / The purpose of this study was to test the antibacterial effect of thin copper treads woven into a polyester fabric. The investigation was done by inoculation of Staphylococcus aureus strain ATCC 6538 to the fabric and evaluation of the number of viable cells post exposure by viable count. The issue to be answered was whether the copper fabric had a bactericide or bacteriostatic effect? The fabric is still in prototype stage, and if proven to have antibacterial properties the aim is to use it to prevent bacterial growth in wounds and other vulnerable locations in clinical care. Copper is an essential trace element, but also has antimicrobial properties through a wide range of mechanisms where cell membrane damage is one of the more important. Methods used for inoculation was the absorption method, where a nutrient broth containing S. aureus was pipetted on to the fabric specimens, and the transfer method where the fabric specimens were pressed onto an agar plate that had previously been spread with peptone salt solution containing S. aureus. Total number of bacteria per fabric specimen after short contact (<1 min) and incubation (18-24 h at 37±2°C) was calculated. Incubation showed significant difference in total number of bacteria between the copper fabric and negative control in three of four tests. Short contact showed a tendency of antibacterial effect. The conclusion was that the copper fabric harmed and killed bacteria during incubation but that more records would be needed to be sure about the effects of short contact on bacteria.

The aim of the study was to evaluate the total viable count and detects each microorganism by the use of mass spectroscopy. In the period of June to December 2017 35 samples of pasteurized milk from two vending machines (Hodkovice, Suchdol) were tested. For comparison samples were divided to warmer season (June-Septemeber) and colder season (October-December) sampling. The total amount of microorganism was determined in each sample of milk and total viable count was estimated by the use of mass spectroscopy MALDI-TOF. The total viable count in the tested pasteurized milk were from 3,27 log KTJ/ml up to 7,35 log KTJ/ml. The mass spectroscopy detected 210 microorganisms belonging to 20 families of gram-positive and gram-negative bacteria and one species of eukaryot organism. The most often identified species belong the Enterobacteriaceae family.

Surface mucus of a freshwater fish, Alburnus alburnus (bleak), caught from Lake Mogan, situated in south of Ankara, was collected in different seasons. The total cultivable bacteria were enumerated by spread plate method on nine different media. Bacteria were isolated based on colony morphologies and pigmentation. A total of sixty bacterial isolates obtained. The mucus-dwelling bacteria were first tested for resistance against ampicillin and kanamycin / then streptomycin and chloramphenicol were added to the experimental set up. The resistance levels of isolates were determined in terms of four antibiotics by tube dilution method. About 90% of the isolates were resistant to chloramphenicol, about 84% to kanamycin, about 88% to streptomycin and about 98% to ampicillin. These high levels of antibiotic resistance are rather interesting from a standpoint that the lake has no record of antibiotics exposure of any sort. The plasmid isolations were carried out to determine if the multiple antibiotic resistance could be attributed to plasmids for starting assumption. But we found no direct relationship between the presence of plasmids and multiple antibiotic resistance. Our study indicated that multiple antibiotic resistance at high levels is among the current phenotypes of the fish mucus-dwelling bacterial populations in Lake Mogan.

The mucus-dwelling bacteria previously isolated from the surface of a freshwater fish species (Alburnus alburnus from Lake Mogan, Ankara), were studied to discover their cobalt resistance. The minimum inhibitory concentrations (MIC) were determined for a total of thirty six bacterial isolates. The results of the resistance studies led us to design experiments on adaptation to cobalt and subsequent memory retention. Three selected isolates were exposed to an inhibitory cobalt concentration as a mixed culture and individually. The delayed formation of colonies along with competitive exclusion of one of the isolates in the mixed culture were recorded. The delay for colony formation was followed up for liquid culture conditions. After some of our isolates acclimated to cobalt and started to exhibit constant time of growth period, it is assumed that they were adapted. We regarded adaptation as a result of memory formation. Next, we did a further study to find out how long this memory could be retained via serial multiple passages in cobalt free medium. We expressed our observations quantitatively by measuring the growth by using spectrophotometer and by performing viable counts. Interestingly, where there was a high CFU, the photometric values were very low. We interpreted the finding such that the presence of cobalt above tolerance limits were causing size reduction in the cells. So that their presence was underestimated by optic devices in visible range. Our study hinted that freshwater bacteria was adapting cobalt in a memory based mechanism and able to retain this memory for some time.