No description available.

ABSTRACT The purpose of this study was to compare the efficiency of three sperm elution methods on sexual assault swabs; factors such as solvent type, solvent volume, sperm concentration, and duration of extraction and elution method were evaluated with respect to observed sperm recovery. Swabs containing dilutions of semen ranging from 1:10 to 1:1,000 and simulated post-coital swabs were extracted via the traditional tube extraction, as well as two direct slide elution techniques, tapping and swirling. For the slide elution techniques, a swab cutting was placed directly onto a microscope slide, a small volume of water or buffer was added, and sperm were eluted by either tapping the sample with a stirring stick or swirling it around the slide with metal forceps. The tube method requires a minimum of one and one half hours for extraction, while the slide elution techniques require only ten seconds for extraction. The average sperm counts from 1:10 dilutions processed with the tapping elution method were statistically higher than the 1:10 dilutions samples processed with tube and swirling methods. Elution by tapping also recovered a significantly higher amount of sperm cells from the 1:1,000 dilution compared to the tube extraction of the same dilution. The tapping elution method consistently resulted in the greatest number of spermatozoa observed, followed by the swirling method and then tube extraction; additionally, incidents of false negatives (no sperm observed) were observed with the tube and swirling methods. Simulated post-coital samples produced similar results to the semen samples; however, vaginal swabs from one donor resulted in an extremely high ratio of exfoliated epithelial cells that obscured the spermatozoa, especially with the direct slide elution methods. The slide elution methods resulted in similar and consistent relative standard deviations between dilutions in samples, while the tube extraction results suggest an increase in variance as the dilution increases. Overall, slide elution methods yielded the most observed sperm cells in a significantly shorter amount of time.

No description available.

Background: The monitoring of infectious diseases like the porcine reproductive and respiratory syndrome (PRRS) using pen-wise oral fluid samples becomes more and more established. The collection of individual oral fluid, which would be useful in the monitoring of PRRSV negative boar studs, is rather difficult. The aim of the study was to test two methods for individual oral fluid collection from pigs and to evaluate the specificity of a commercial ELISA for detection of PRRSV antibodies in these sample matrices. For this reason, 334 serum samples from PRRSV negative pigs (group 1) and 71 serum samples from PRRSV positive pigs (group 2) were tested for PRRSV antibodies with a commercial ELISA. Individual oral fluid was collected with a cotton gauze swab from 311 pigs from group 1 and 39 pigs from group 2. Furthermore, 312 oral fluid samples from group 1 and 67 oral fluid samples from group 2 were taken with a self-drying foam swab (GenoTube). The recollected oral fluid was then analysed twice with a commercial ELISA for detection of PRRSV antibodies in oral fluid.

Quatro tipos de meios de transporte comerciais (Amies, Amies com carvão, Cary Blair e Stuart) e suabes sem meio foram avaliados quanto à capacidade de conservar S. hyicus por 10 dias de armazenamento sob duas temperaturas (ambiente [20-25ºC] e refrigerada [4-8ºC]). Não foram encontrados na literatura trabalhos que avaliassem o efeito da temperatura e de meios de transporte na sobrevivência desta bactéria. Foi usado o Roll-Plate Method para o teste, utilizando uma amostra padrão de S. hyicus (ATCC 11249), de acordo com as normas do NCCLS (2003). Durante o período de 10 dias, as amostras foram armazenadas nas duas temperaturas e a seguir os suabes foram plaqueados em Ágar Tween 80 com contagem das colônias a cada 24 horas. As amostras conservadas em todos os meios de transporte apresentaram desempenho significativamente superior (P ≤ 0,05) quando mantidas em refrigeração. A conservação no Amies refrigerado foi superior a todos os outros tipos de meios e suabes (P ≤ 0,05). Na temperatura ambiente, o meio Amies e os suabes mantidos sem meio de transporte foram similares (P > 0,05), sendo os melhores nesta temperatura. Adicionalmente, o meio Amies refrigerado demonstrou alta performance até nove dias de armazenamento e os suabes sem meio até três dias. Coletivamente, a alta performance em meio Amies refrigerado indicou ser este o mais adequado para o transporte de amostras de S. hyicus. No entanto, devido à resistência à dessecação da bactéria e em função da praticidade e baixo custo, poderiam também ser indicados suabes sem meio de transporte, mantidos na temperatura ambiente. Esta forma de envio seria viável quando a previsão de tempo para o transporte ao laboratório for menor que 48 horas, pois nesta forma de conservação houve sobrevivência adequada até 72 horas. / Four commercial transport media (Amies, Amies with charcoal, Cary Blair and Stuart) and swabs without transport medium were assessed regarding their capacity to preserve S. hyicus for periods of up to 10 days at two different temperatures (room temperature [20-25ºC] and refrigerated [4-8ºC]). We did not find in the literature information on the effect of temperature and transport media for the survival of this bacteria. The Roll-Plate Method was used with a reference strain of S. hyicus (ATCC 11249), following recommendation of NCCLS (2003). The samples were stored for 10 days in two temperatures and after that, swabs were plated in Tween 80 Agar, counting bacterial colonies at each 24 hours. Samples held in all transport media showed significantly better performance (P ≤ 0.05) in refrigeration. Storage in refrigeration in Amies medium performed better than all other transport media and swabs (P ≤ 0.05). In room temperature, Amies medium and swabs without transport medium worked similarly (P > 0.05), and presented the bests results in this temperature. Additionally, refrigerated Amies medium showed high performance for up to nine storage days and swabs without transport medium for up to three days. Collectively, the high performance in refrigerated Amies medium indicated that this is the most suitable medium for shipping of S. hyicus. However, considering S. hyicus resistance to desiccation, easiness of transportation and low cost, swabs without transport medium could also be indicated, when stored at room temperature. This form of shipment could be viable when the time predicted to transport to the laboratory would be smaller than 48 hours, since this form of preservation showed survival for up to 72 hours.

Quatro tipos de meios de transporte comerciais (Amies, Amies com carvão, Cary Blair e Stuart) e suabes sem meio foram avaliados quanto à capacidade de conservar S. hyicus por 10 dias de armazenamento sob duas temperaturas (ambiente [20-25ºC] e refrigerada [4-8ºC]). Não foram encontrados na literatura trabalhos que avaliassem o efeito da temperatura e de meios de transporte na sobrevivência desta bactéria. Foi usado o Roll-Plate Method para o teste, utilizando uma amostra padrão de S. hyicus (ATCC 11249), de acordo com as normas do NCCLS (2003). Durante o período de 10 dias, as amostras foram armazenadas nas duas temperaturas e a seguir os suabes foram plaqueados em Ágar Tween 80 com contagem das colônias a cada 24 horas. As amostras conservadas em todos os meios de transporte apresentaram desempenho significativamente superior (P ≤ 0,05) quando mantidas em refrigeração. A conservação no Amies refrigerado foi superior a todos os outros tipos de meios e suabes (P ≤ 0,05). Na temperatura ambiente, o meio Amies e os suabes mantidos sem meio de transporte foram similares (P > 0,05), sendo os melhores nesta temperatura. Adicionalmente, o meio Amies refrigerado demonstrou alta performance até nove dias de armazenamento e os suabes sem meio até três dias. Coletivamente, a alta performance em meio Amies refrigerado indicou ser este o mais adequado para o transporte de amostras de S. hyicus. No entanto, devido à resistência à dessecação da bactéria e em função da praticidade e baixo custo, poderiam também ser indicados suabes sem meio de transporte, mantidos na temperatura ambiente. Esta forma de envio seria viável quando a previsão de tempo para o transporte ao laboratório for menor que 48 horas, pois nesta forma de conservação houve sobrevivência adequada até 72 horas. / Four commercial transport media (Amies, Amies with charcoal, Cary Blair and Stuart) and swabs without transport medium were assessed regarding their capacity to preserve S. hyicus for periods of up to 10 days at two different temperatures (room temperature [20-25ºC] and refrigerated [4-8ºC]). We did not find in the literature information on the effect of temperature and transport media for the survival of this bacteria. The Roll-Plate Method was used with a reference strain of S. hyicus (ATCC 11249), following recommendation of NCCLS (2003). The samples were stored for 10 days in two temperatures and after that, swabs were plated in Tween 80 Agar, counting bacterial colonies at each 24 hours. Samples held in all transport media showed significantly better performance (P ≤ 0.05) in refrigeration. Storage in refrigeration in Amies medium performed better than all other transport media and swabs (P ≤ 0.05). In room temperature, Amies medium and swabs without transport medium worked similarly (P > 0.05), and presented the bests results in this temperature. Additionally, refrigerated Amies medium showed high performance for up to nine storage days and swabs without transport medium for up to three days. Collectively, the high performance in refrigerated Amies medium indicated that this is the most suitable medium for shipping of S. hyicus. However, considering S. hyicus resistance to desiccation, easiness of transportation and low cost, swabs without transport medium could also be indicated, when stored at room temperature. This form of shipment could be viable when the time predicted to transport to the laboratory would be smaller than 48 hours, since this form of preservation showed survival for up to 72 hours.

Quatro tipos de meios de transporte comerciais (Amies, Amies com carvão, Cary Blair e Stuart) e suabes sem meio foram avaliados quanto à capacidade de conservar S. hyicus por 10 dias de armazenamento sob duas temperaturas (ambiente [20-25ºC] e refrigerada [4-8ºC]). Não foram encontrados na literatura trabalhos que avaliassem o efeito da temperatura e de meios de transporte na sobrevivência desta bactéria. Foi usado o Roll-Plate Method para o teste, utilizando uma amostra padrão de S. hyicus (ATCC 11249), de acordo com as normas do NCCLS (2003). Durante o período de 10 dias, as amostras foram armazenadas nas duas temperaturas e a seguir os suabes foram plaqueados em Ágar Tween 80 com contagem das colônias a cada 24 horas. As amostras conservadas em todos os meios de transporte apresentaram desempenho significativamente superior (P ≤ 0,05) quando mantidas em refrigeração. A conservação no Amies refrigerado foi superior a todos os outros tipos de meios e suabes (P ≤ 0,05). Na temperatura ambiente, o meio Amies e os suabes mantidos sem meio de transporte foram similares (P > 0,05), sendo os melhores nesta temperatura. Adicionalmente, o meio Amies refrigerado demonstrou alta performance até nove dias de armazenamento e os suabes sem meio até três dias. Coletivamente, a alta performance em meio Amies refrigerado indicou ser este o mais adequado para o transporte de amostras de S. hyicus. No entanto, devido à resistência à dessecação da bactéria e em função da praticidade e baixo custo, poderiam também ser indicados suabes sem meio de transporte, mantidos na temperatura ambiente. Esta forma de envio seria viável quando a previsão de tempo para o transporte ao laboratório for menor que 48 horas, pois nesta forma de conservação houve sobrevivência adequada até 72 horas. / Four commercial transport media (Amies, Amies with charcoal, Cary Blair and Stuart) and swabs without transport medium were assessed regarding their capacity to preserve S. hyicus for periods of up to 10 days at two different temperatures (room temperature [20-25ºC] and refrigerated [4-8ºC]). We did not find in the literature information on the effect of temperature and transport media for the survival of this bacteria. The Roll-Plate Method was used with a reference strain of S. hyicus (ATCC 11249), following recommendation of NCCLS (2003). The samples were stored for 10 days in two temperatures and after that, swabs were plated in Tween 80 Agar, counting bacterial colonies at each 24 hours. Samples held in all transport media showed significantly better performance (P ≤ 0.05) in refrigeration. Storage in refrigeration in Amies medium performed better than all other transport media and swabs (P ≤ 0.05). In room temperature, Amies medium and swabs without transport medium worked similarly (P > 0.05), and presented the bests results in this temperature. Additionally, refrigerated Amies medium showed high performance for up to nine storage days and swabs without transport medium for up to three days. Collectively, the high performance in refrigerated Amies medium indicated that this is the most suitable medium for shipping of S. hyicus. However, considering S. hyicus resistance to desiccation, easiness of transportation and low cost, swabs without transport medium could also be indicated, when stored at room temperature. This form of shipment could be viable when the time predicted to transport to the laboratory would be smaller than 48 hours, since this form of preservation showed survival for up to 72 hours.

No description available.

No description available.

[Truncated abstract] The collection of buccal cells is common practise in the epidemiological and forensic science. Unlike venipuncture collection of blood; it is a safer, non-invasive method for collection of biological material. The methods by which these cells are collected from the inner cheek of an individual and stored are the key elements in preserving DNA. Typically, forensic samples require long term storage. Samples are commonly collected on cotton swabs and stored moist at low to ultra-low temperatures (less than -20oC). Although this is the method of choice in most forensic facilities, there are drawbacks. The samples are inherently contaminated with microflora within the oral cavity and the moisture allows a plethora of microorganisms to grow. As the time frame that has elapsed from collection to storage increases, there is an exponential increase in bacterial cells. Storage of containers containing swabs coated with cells at temperatures below 20oC is also costly due to requirements for large freezers which are running and monitored over 24 hours. In the pass 10 to 15 years, researchers have focussed on alternative ways to store buccal cells. The FTA card system by Whatman is one such development. The FTA card is unique in that it provides a means for the collection of buccal cells for storage at room temperature. DNA profiling from samples stored in this way for 11 years has been successfully achieved. The filter paper matrix of the FTA card binds and subsequently lyses cells. ... (2) The second component of this thesis describes a study which subjected cells on buccal swabs to various conditions of increased temperature over periods of time to establish if DNA could be amplified. The aim was to mimic exposure to the vigours of field conditions, particularly in the extreme local environments that prevail in the United Arab Emirates. a. Initially, buccal cells stored at -20oC over 360 days were used to mimic standard archiving procedures. The cells were subsequently transferred to FTA cards, amplified and profiled by using ABI AmpFLSTR Identifiler PCR Amplification Kit (Applied Biosystems, Foster City, CA). Complete STR profiles were successfully recovered from the archived swabs. In most cases 100% of alleles were recovered, suggesting that it is feasible to transfer DNA from properly archived buccal swabs to FTA cards. b. The second phase involved the storage of fresh swabs that had been artificially aged by using incubation temperatures ranging from 40oC to 100oC. Partial profiles resulted from artificially aged samples, indicating that the prevailing conditions prior to low temperature storage of the swabs plays an important role in ensuring cellular integrity and thus, DNA quality. Results from this study suggest that it is possible for biological samples stored under correct conditions to be transferred from swabs to FTA card. In combination, the two chapters presented in this study show that it is feasible to transfer achieved forensic biology samples from swabs to the FTA card system. However, it is necessary to ensure that the samples are treated in the correct manner so as to minimise contamination from external sources and to maintain the correct environmental state to maintain intact cells and usable DNA.