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Direct immunofluorescence is a vital diagnostic test for assessing vesiculobullous disorders, vasculitides, and connective tissue diseases. It is a robust and valuable technique that offers essential diagnostic information for many critical dermatoses. Dermatopathologists depend heavily on the data obtained from direct immunofluorescence evaluation to confirm final diagnoses. Selecting the most appropriate biopsy site is necessary for maximizing diagnostic accuracy, and the best site may vary depending on the clinical differential diagnosis. Inaccurate biopsy site selection can significantly impact the accuracy of the results. To optimize the use of direct immunofluorescence studies, this review provides helpful guidelines and some practical tips for selecting the best biopsy site. [ABSTRACT FROM AUTHOR]

Introduction: Avulsion of teeth is one of the most complex forms of dental injury, and the selection of an appropriate storage medium greatly influences the preservation of Periodontal Ligament (PDL) cell viability, which is crucial for the successful re-implantation of avulsed teeth. Therefore, identifying effective storage options such as natural coconut water and prepackaged coconut water holds significant promise in improving outcomes for this challenging dental injury. Aim: To evaluate the efficacy of natural coconut water, prepackaged coconut water, and Hank's Balanced Salt Solution (HBSS) as storage media in maintaining PDL cell viability. Materials and Methods: A total of 32 non-carious freshly extracted human premolars were randomly divided into four study groups (n=8) and stored in the following storage media respectively: Group I-Natural coconut water group, Group IIPre-packaged coconut water group, Group III-Bench dry group, and Group IV-HBSS groups for 30 minutes. The PDL cells were collected and incubated in phosphate buffer saline for 30 minutes and then centrifuged at 800 rpm for five minutes. Following this, the cells were stained with trypan blue to observe their viability. The Analysis of Variance (ANOVA) with Tukey's post-hoc test was used for analysing the data. Results: The mean percentage of viable cells in natural coconut water (80.6250) was higher than in HBSS (79.8750), prepackaged coconut water (79.2500), and the bench dry group (6.1250). Meanwhile, the mean percentage of non-viable cells was highest in the bench dry group (93.8750), followed by the pre-packaged coconut water (20.7500), HBSS (20.1250), and natural coconut water (19.3750). Conclusion: Natural coconut water and pre-packaged coconut water are equally effective in maintaining the viability of PDL cells. Therefore, pre-packaged coconut water can be used as a substitute for natural coconut water for tooth storage, depending upon availability. [ABSTRACT FROM AUTHOR]

Background: Neisseria gonorrhoeae (Gonococcus-GC) is a fastidious, autolytic Gram-negative diplococcus with stringent growth requirements, and cannot be cultivated in a routine microbiology laboratory, without well-equipped incubators, reagents, and special media. Hence, many clinics and laboratories prefer to ship the specimens or isolates to a dedicated referral laboratory for confirmation of isolates and antimicrobial susceptibility testing. Thus, transportation conditions for gonococcal isolates, become crucial for its recovery and successful isolation in the laboratory. This retrospective study was conducted at a national referral laboratory for gonococcus, in India, over 3 years. Aim: In this study, an attempt was made to determine the factors affecting the revival of isolates of gonococci, that were despatched, from across India, to the referral laboratory for confirmation of species and antimicrobial susceptibility patterns. Method: Over 3 years, the culture plates, test tubes, or vials used for transporting gonococcal isolates, and their modes of transport to the referral laboratory, were studied in detail. The isolates were revived (whenever possible), subcultured, and identified by standard methods in the referral laboratory. Results: A total of 77 samples were processed for revival and 83.12% of isolates were recovered, with failure of recovery in 16.88% of specimens. Conclusion: Several factors play a role in the successful revival of N. gonorrhoeae from culture isolates transported across the Indian subcontinent. These include purity of growth, culture media used for transport, sending of isolates in duplicates, temperature, time, distance, and season of transport. All these factors must be kept in mind when transporting gonococcal isolates, for successful revival. Finally, the skills of the laboratory technician are of immense importance too. [ABSTRACT FROM AUTHOR]

Introduction: The sudden arrival of the COVID-19 pandemic placed significant stresses on supply chains including viral transport medium (VTM). The VTM that was urgently required needed to support viral replication, as well as other routine diagnostic approaches. We describe the preparation and validation testing of VTM for rapidly expanding diagnostic testing, where the capacity of the VTM to preserve viral integrity, for culture, isolation and full sequence analysis, was maintained. Methods: VTM was prepared using different methods of sterilization then 'spiked' with virus. The VTM was investigated using viral culture in Vero cells, and for nucleic acid detection by quantitative PCR. Results: The best results were obtained by filter and autoclave-based sterilization. The VTM proved robust for culture-based analyses provided the inoculated VTM was stored at 4 °C, and tested within 48 h. The filtered VTM also supported PCR-based diagnosis for at least 5 days when the mock inoculated VTM was held at room temperature. Discussion: The manual handling of VTM production, including filling and sterilization, was optimized. SARS-CoV-2 was spiked into VTM to assess different sterilization methods and measure the effects of storage time and temperature upon VTM performance. While most diagnostic protocols will not require replication competent virus, the use of high quality VTM will allow for the next phase of laboratory analysis in the COVID-19 pandemic, including drug and antibody susceptibility analysis of re-isolated SARS-CoV-2, and for the testing of vaccine escape mutants. [ABSTRACT FROM AUTHOR]

Background and Objectives: Antimicrobial resistance of Neisseria gonorrhoeae is globally spread and threatening. Culturing of N. gonorrhoeae is the only method to collect live isolates for investigation antimicrobial resistance profile. Therefore, quality assessment of N. gonorrhoeae culture is essential for successful isolation of gonococci. This study was conducted to evaluate deferred and bedside culture of N. gonorrhoeae depending on the year season and temperature condition of transport media temporary storage. Materials and Methods: Urogenital swabs from 46 symptomatic heterosexual patients with gonorrhoea and subculture of N. gonorrhoeae in 46 suspensions in concentrations 1.5 × 108 CFU/ml were subjected to the study. Non-nutritive transporting medium Amies Agar Gel Medium with charcoal (Copan Diagnostics Inc., Brescia, Italy) was used for deferred culture and selective Chocolate agar TM+PolyViteX VCAT3 (BioMérieux, Marcy-l'Étoile, France) for both tested methods of culture. Results: The specificity of both bedside and deferred methods of culture was 100%. The sensitivity of deferred culture was higher than of bedside culture (82.6% vs 47.8%, p<0.0005). Deferred culture showed significantly higher sensitivity comparing to bedside culture in summer (100% vs 50%, p=0.003), and comparably the same as for bedside culture in autumn, winter and spring. Conclusion: The viability of N. gonorrhoeae subcultures was significantly higher in refrigerated samples from transport media than from ambient one after exposition from 48 to 96 hours. Optimal viability of N. gonorrhoeae was observed when transport swabs were kept refrigerated up to 48 h (73.9-93.5%) or ambiently - up to 24 h (87%). Updating laboratory guidelines regarding sampling and timely specimen processing might improve gonococcal culture performance. [ABSTRACT FROM AUTHOR]

Introduction: Collecting swabs from skin lesions for bacteriological examination is frequently performed to the diagnosis of exudative epidermitis. This method is fast and non-invasive, but it depends directly on the viability of bacteria in clinical samples, which can be influenced by storage and shipment temperatures and the time of transportation. The aim of this study was to assess the capacity of four commercial transport media and swabs with no transport medium to preserve Staphylococcus hyicus (S. hyicus) for up to 10 days at room temperature and under refrigeration. Methodology: Samples were stored in swabs with no transport medium and four transport media (Amies, Amies with charcoal, Cary Blair and Stuart) for 10 days at room temperature and under refrigeration. Swabs were plated in Tween 80 Agar and colonies counted. Results: Samples kept in transport media showed better performance (P < 0.05) under refrigeration. Storage under refrigeration in Amies medium showed better results than all other transport media and swabs (P < 0.05). Amies medium and swabs with no transport medium showed comparable results in room temperature (P > 0.05). In additional, refrigerated Amies medium and swabs with no transport medium at room temperature showed high performance for up to nine and three storage days, respectively. Conclusions: The recovery of S. hyicus in samples stored in Amies medium under refrigeration was higher when compared to other transport media. In addition, swabs with no transport medium could also be indicated when samples are stored at room temperature within three days. [ABSTRACT FROM AUTHOR]

Brachyspira (B.) hyodysenteriae is a causative agent of swine dysentery that is responsible for death and economic losses in the pig industry. It is imperative that clinical samples be delivered fresh for accurate diagnosis. The viability and DNA detection of B. hyodysenteriae using lab-made (phosphate buffered saline and modified tryptic soy broth) or commercial transport media (C, D, and E) were compared by culturing and real-time PCR at 4oC or room temperature (RT), respectively. B. hyodysenteriae grown in D (Anaerobe Systems, USA) and E (Starplex Scientific, Canada) media was viable for 4 days at 4oC and RT. However, B. hyodysenteriae in A, B, and C (culture swab; BD Biosciences, USA) media were not recovered after 2 days at RT. Ct values for real-time PCR at 4oC and RT ranged from 27.2 ± 2.1 (C) to 29.6 ± 0.5 (B), and 28.0 ± 0.9 (E) to 30.2 ± 1.5 (B), respectively. Considering the field conditions, it is important that transport media is used for specimen isolation and PCR to obtain an accurate diagnosis of swine dysentery. [ABSTRACT FROM AUTHOR]

Background: Accurate microbial diagnosis is crucial for effective management of prosthetic joint infections. Culturing of multiple intraoperative tissue samples has increased diagnostic accuracy, but new preparatory techniques and molecular methods hold promise of further improvement. The increased complexity of sampling is, however, a tough challenge for surgeons and assistants in the operation theatre, and therefore we devised and tested a new concept of pre-packed boxes with a complete assortment of swabs, vials and additional tools needed in the operating theatre for non-standard samples during a clinical study of prosthetic joint infections. Findings: The protocol for the clinical study required triplicate samples of joint fluid, periprosthetic tissue, bone tissue, and swabs from the surface of the prosthesis. Separate boxes were prepared for percutaneous joint puncture and surgical revision; the latter included containers for prosthetic components or the entire prosthesis. During a 2-year project period 164 boxes were used by the surgeons, 98 of which contained a complete set of samples. In all, 1508 (89%) of 1685 scheduled samples were received. Conclusion: With this concept a high level of completeness of sample sets was achieved and thus secured a valid basis for evaluation of new diagnostics. Although enthusiasm for the project may have been a contributing factor, the extended project period suggests that the ‘All in a box’ concept is equally applicable in routine clinical settings with standardized but complex diagnostic sampling. [ABSTRACT FROM AUTHOR]

Abstract: The Government Accountability Office report investigating the surface sampling methods used during the 2001 mail contamination with Bacillus anthracis brought to light certain knowledge gaps that existed regarding environmental sampling with biothreat agents. Should a contamination event occur that involves non-spore forming biological select agents, such as Yersinia pestis, surface sample collection and processing protocols specific for these organisms will be needed. Two Y. pestis strains (virulent and avirulent), four swab types (polyester, macrofoam, rayon, and cotton), two pre-moistening solutions, six transport media, three temperatures, two levels of organic load, and four processing methods (vortexing, sonicating, combined sonicating and vortexing, no agitation) were evaluated to determine the conditions that would yield the highest percent of cultivable Y. pestis cells after storage. The optimum pre-moistening agent/transport media combination varied with the Y. pestis strain and swab type. Directly inoculated macrofoam swabs released the highest percent of cells into solution (93.9% recovered by culture) and rayon swabs were considered the second best swab option (77.0% recovered by culture). Storage at 4°C was found to be optimum for all storage times and transport media. In a worst case scenario, where the Y. pestis strain is not known and sample processing and analyses could not occur until 72h after sampling, macrofoam swabs pre-moistened with PBS supplemented with 0.05% Triton X-100 (PBSTX), stored at 4°C in neutralizing buffer (NB) as a transport medium (PBSTX/NB) or pre-moistened with NB and stored in PBSTX as a transport medium (NB/PBSTX), then vortexed 3min in the transport medium, performed significantly better than all other conditions for macrofoam swabs, regardless of strain tested (mean 12 – 72h recovery of 85.9–105.1%, p <0.001). In the same scenario, two combinations of pre-moistening medium/transport medium were found to be optimal for rayon swabs stored at 4°C (p <0.001), then sonicated 3min in the transport medium; PBSTX/PBSTX and NB/PBSTX (mean 12–72h recovery of 83.7–110.1%). [Copyright &y& Elsevier]

Objective: Both the length of extra-alveolar time and type of storage media are significant factors that can affect the long-term prognosis of replanted teeth. This study aims to compare propolis 50%, propolis 10%, Hank's balanced salt solution (HBSS), milk and egg white on periodontal ligament (PDL) cell survival for different time points. Materials and Methods: In this in vitro experimental study, we divided 60 extracted teeth without any periodontal diseases into five experimental and two control groups that consisted each experimental group with 10 and each control group with 5 teeth. The storage times were one and three hours for each media. The controls corresponded to 0-minute (positive) and 12-hour (negative) dry time. Rinsing in the experimental media, the teeth were treated with dispase and collagenase for one hour. Cell viability was determined by using trypan blue exclusion. Statistical analysis of the data was accomplished by using two-way analysis of variance (ANOVA) complemented by the Tukey's HSD post-hoc. Results: Within one hour, there was no significant difference between the two propolis groups, however these two groups had significantly more viable PDL cells compared to the other experimental media (p<0.05). The results of the three-hour group showed that propolis 10% was significantly better than egg white, whereas both propolis 10% and 50% were significantly better than milk (p<0.05). Conclusion: Based on PDL cell viability, propolis could be recommended as a suitable biological storage media for avulsed teeth. [ABSTRACT FROM AUTHOR]

Objective: To evaluate group B streptococcus (GBS) detection in an in vitro setting, using a low and controlled inoculum from swabs directly inoculated into a selective medium, as compared to delayed inoculation following a period in a commercial Amies transport medium with charcoal (Venturi Transystem™ Copan, Italy). Study design: Clinical isolates of GBS (n = 103), were inoculated into the Amies transport medium with charcoal in a concentration of 100 colony-forming units (cfu)/ml (10 cfu/swab). Swabs were then transferred to an enrichment broth (NPC) at time intervals of 0, 2, 4, 6 and 24 hours. Broths were then incubated for 18-24 hours at 35°C in air, before being transferred to New Granada Medium Modified (NGM) for CBS detection and incubated for a further 18-24 hours at 35°C in air. If the characteristic orange pigmented colonies were observed after this period, the specimen was recorded as + (1-10 colonies) or + + (more than 10 colonies). Results: Overall 92.2% (95/103) of isolates were detected in all tubes and at all times. An additional two isolates were non-hemolytic, non-pigment forming GBS. Of note, 3.9% (4/103) were negative until 2 hours delayed inoculation and 1.9% (2/103) gave inconsistent results, likely due to the low inoculum used. Conclusion: Delayed inoculation into selective enrichment broth following a period in transport medium, even with a low inoculum, gave a similar and acceptable GBS detection rate to direct inoculation. Hence, Amies transport medium with charcoal is an appropriate transport medium to use, where it is not practical for clinical specimens to be directly inoculated into selective enrichment broth and as endorsed in the Centers for Diseases Control (CDC) Guidelines, 2002. [ABSTRACT FROM AUTHOR]

The forthcoming MPEG-4 standard specifies in its systems part an audiovisual scene description and functionality for the elementary stream management. The elementary stream management functionality is introduced. It consists of a media object description framework that describes the streaming resources that form part of an MPEG-4 presentation and of a synchronization syntax incorporated in a flexible sync layer with an underlying systems decoder model. The final section outlines the transport and session setup for MPEG-4 presentations on relevant transport media, namely, the Internet and in digital broadcast scenarios [ABSTRACT FROM PUBLISHER]

The purpose of this study was to evaluate the survival in 3 transport media of 3 suspected periodontal pathogens, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia. Subgingival samples were taken from 10 patients with severe periodontitis, all harboring at least two of the above- mentioned species. The material was dispersed and aliquots were added to vials containing reduced transport fluid, reduced transport fluid containing 10% Fildes extract, or viability-maintaining microbiostatic medium, anaerobically pre- pared (VMGA Ill). Viable counts were determined after 1, 2, 4, 24 and 48 h of storage at 4°C or at room temperature. The results showed that, for up to 4 h of storage, no significant differences existed for all parameters tested. A large increase of the total viable counts was found in VMGA III at room temperature after 24 and 48 h. This was due to an outgrowth of mainly streptococci. Incubation at 4°C yielded often a significantly higher recovery compared to room temperature. After storage at room temperature, the tested bacteria were below detection level in some samples. [ABSTRACT FROM AUTHOR]

This study focused on several factors that may influence the percentage of motile microorganisms in a sample from the tongue determined using a phase-contrast microscope. It was found that the time elapsing between sampling and analysis is crucially important. In both sterile saline and reduced transport fluid (RTF), a reduction of the percentage of motile microorganisms was found within 15-30 min after sampling respectively. In order to be able to prolong the time interval between sampling and analysis, it was found that storing a sample in a tuberculin syringe with RTF supplemented with Fildes extract preserved the motility of the microorganisms and gave no significant reduction in the percentage of motiles within 48 h. [ABSTRACT FROM AUTHOR]

Abstract: Background/Aims: A wide variety of materials has been researched for their use as potential storage media for avulsed teeth, but it is essential to recognize the medium most recommended for improvement of the prognosis of avulsed teeth. The aim of this systematic review was to identify the most recommended medium to store and transport avulsed teeth based on the survival of periodontal ligament (PDL) cells as determined by in vitro studies. Methods: Only laboratory‐based experimental studies on PDL cells found on adult permanent teeth were included. Data were collected using PubMed, CINAHL plus (EBSCO host), and the Cochrane Library, along with Google Scholar and a hand search. The key terms employed were permutations of [avulsed permanent teeth* OR dental avulsion* OR knocked out teeth*] AND [storage media* OR transport media* OR biological transport* OR PDL cell viability* OR PDL cell survival*]. A customized data extraction pro forma was used to extract the data and to evaluate the quality and risk of bias. Results: The initial search yielded 978 articles, but only 67 were selected. Milk was the most recommended individual medium followed by Hank's balanced salt solution. Among natural products other than milk, propolis and coconut water were most frequently recommended. Recommendations were based on maintenance of PDL cell viability followed by ease of availability, low cost, and long shelf life. Conclusions: Natural products are more effective in maintaining the PDL cell viability compared to synthetic products. Some storage media recommendations were also based upon practical aspects. Although natural products other than milk have more recommendations as a group, milk is the most recommended storage medium individually, based not only on PDL cell viability, but also practical considerations. [ABSTRACT FROM AUTHOR]