agar and Cadaver

In cadavers, even Thiel-embalmed cadavers, the arteries (especially the thoracic aorta) are extremely collapsed. This is in marked contrast to the state of the arteries in a living body. Aortic inflation is necessary to improve this unfavorable situation for anatomical observation or dissection. To inflate the aorta, we injected 500 ml of hot liquid agar into the aorta using a 18-Fr catheter inserted into the common femoral artery and subclavian artery. The injected agar then rapidly cools to room temperature and solidifies. As a result, the thoracic aorta remains sufficiently and constantly inflated in the mediastinum. This method is not only easy and inexpensive, but also useful and effective for achieving a life-like anatomy in cadavers used in surgical training for operations involving mediastinal organs, with the exception of the heart and great vessels.

Topics: Agar; Aorta, Thoracic; Cadaver; Embalming; Humans; Mediastinum; Thoracic Surgery

Ultra-structural analysis of human post-mortem brain tissue is important for investigations into the pathomechanism of neuropsychiatric disorders, especially those lacking alternative models of studying human-specific morphological features. For example, Von Economo Neurons (VENs) mainly located in the anterior cingulate cortex and in the anterior part of the insula, which seem to play a role in a variety of neuropsychiatric conditions, including frontotemporal dementia, autism and schizophrenia, can hardly be studied in nonhuman animals. Accordingly, little is known about the ultra-structural alterations of these neurons, though important research using qualitative stereological methods has revealed that protein expression of the VENs assigns them a role in immune function. Formaldehyde, which is the most common fixative in human pathology, interferes with the immunoreactivity of the tissue, possibly leading to unreliable results. Therefore, a method for ultra-structural investigations independent of antigenic properties of the fixated tissue is needed. Here, we propose an approach using electron microscopy to examine cytoskeletal structures, synapses and mitochondria in these cells. We also show that our methodology is able to keep tissue consumption to a minimum, while still allowing for the specimens to be handled with ease by using agar embedded slices in contrast to blocks for the embedding procedure. Accordingly, a stepwise protocol utilising 60μm thick human post mortem brain sections for electron microscopic ultra-structural investigations is presented.

Topics: Agar; Cadaver; Fixatives; Formaldehyde; Gyrus Cinguli; Humans; Microscopy, Electron; Neurons; Parkinson Disease, Postencephalitic; Specimen Handling; Tissue Embedding

As part of the donor assessment protocol, bioburden assessment must be performed on allograft musculoskeletal tissue samples collected at the time of tissue retrieval. Swab samples of musculoskeletal tissue allografts from cadaveric donors are received at the microbiology department of the South Eastern Area Laboratory Services (Australia) to determine the presence of bacteria and fungi. This study will review the isolation rate of organisms from solid agar and broth culture of swab samples of cadaveric allograft musculoskeletal tissue over a 6-year period, 2006-2011. Swabs were inoculated onto horse blood agar (anaerobic, 35 °C) and chocolate agar (CO2, 35 °C) and then placed into a cooked meat broth (aerobic, 35 °C). A total of 1,912 swabs from 389 donors were received during the study period. 557 (29.1 %) swabs were culture positive with the isolation of 713 organisms, 249 (34.9 %) from solid agar culture and an additional 464 (65.1 %) from broth culture only. This study has shown that the broth culture of cadaveric allograft musculoskeletal swab samples recovered a greater amount of organisms than solid agar culture. Isolates such as Clostridium species and Staphylococcus aureus would not have been isolated from solid agar culture alone. Broth culture is an essential part of the bioburden assessment protocol of swab samples of cadaveric allograft musculoskeletal tissue in this laboratory.

Topics: Agar; Allografts; Bacteria; Cadaver; Culture Media; Humans; Musculoskeletal System

We previously applied our method of detecting marine or freshwater bacterioplankton (bacteria) in the blood of immersed victims as a marker of drowning. However, we did not confirm the absence of post-mortem bacterial invasion during immersion. Here we examined the nature of bacterioplankton in blood samples from 21 immersed and 4 non-immersed cadavers. We found only freshwater bacterioplankton in the blood of two victims that were retrieved from the sea or an estuary inhabited by marine bacterioplankton even though one victim was highly putrefied. The results of diatom testing suggested that these two victims had drowned in fresh or brackish water with low salinity and then flowed out to the estuary or the sea. Two others were submerged in water, but representative bacterioplankton were undetectable in their blood although one victim was highly putrefied. Autopsy findings and the results of diatom tests did not indicate that the cause of death was drowning. As in previous studies, we identified freshwater bacterioplankton in the blood of seven other victims that had drowned in freshwater, marine bacterioplankton in the blood of four victims that had drowned in seawater and none in four victims found on land that had died by means other than drowning. Bacterioplankton in the blood of drowned victims appears to reflect the type of water aspirated and blood does not become easily contaminated with bacteria post-mortem even in decomposed bodies.

Topics: Adult; Agar; Aged; Aged, 80 and over; Animals; Bacteria; Cadaver; Culture Media; Diatoms; Drowning; Forensic Pathology; Fresh Water; Humans; Immersion; Kidney; Liver; Lung; Middle Aged; Plankton; Postmortem Changes; Rivers; Seawater

We measured bacterioplankton in blood from cadavers retrieved from the sea (n=12), near estuaries (n=4), rivers (fresh water, n=8) and from bathtubs (n=4) as well as from non-drowned victims (n=10) discovered near aquatic environments. Blood from 11 victims drowned in seawater developed bioluminescent and/or blue colonies (oxidase test positive) on selective media containing 2-4% NaCl. Homology analyses of the 16S rRNA gene showed that all of them were marine bacteria (genera: Photobacterium, Vibrio, Shewanella, Psychrobacter). Blood from all victims drowned in rivers generated blue colonies on plates containing 3%, but not 4% NaCl. Homology analyses showed that the blue colonies were generated from bacteria that inhabit fresh water (Aeromonas). None of the blood samples from victims that drowned in bathtubs generated bioluminescent and blue colonies. However, all cadavers contained bacteria that produced unstained colonies (Staphylococcus, Bacillus, Enterobacter, Escherichia, etc.). Among non-drowned victims, blood from two gave rise to blue colonies on plates containing < or =3% NaCl (Pseudomonas). Of the cadavers found near estuaries, bioluminescent and blue colonies developed from two of them on media containing 2-4% NaCl (Photobacterium, Vibrio, Listonella), but not from two others on plates containing 4% NaCl (at < or =3%; blue colonies, Aeromonas; unstained colonies, Citrobacter, Vagococcus, Proteus, Enterobacter). These results suggested that the presence of numerous bacterioplankton in immersed cadavers could support a conclusion of death by drowning.

Topics: Agar; Bacteria; Baths; Cadaver; Case-Control Studies; Culture Media; Drowning; Forensic Pathology; Fresh Water; Humans; Immersion; Luminescence; Plankton; RNA, Ribosomal, 16S; Seawater

To investigate the effectiveness of marine bacteria as a new marker of drowning in seawater, we determined the optimal conditions of media required to selectively detect marine bacteria and applied the technique to drowned cadavers. We incubated model blood samples (n=20 per group) mixed with seawater, river, tap or muddy water on agar plates (Todd Hewitt, TH; Marine 2216, M2216) and determined the NaCl concentration required to selectively detect marine bacteria. We also used TCBS agar plates without manipulation to isolate Vibrio spp. Among the culture media, TH agar was superior. Bioluminescent colonies were detected only in blood mixed with seawater. Blue colonies stained using the cytochrome oxidase test (COT), were detected in blood mixed with both sea and river water. However when the NaCl concentration was above 4%, COT stained colonies were detectable only in blood mixed with seawater. We subsequently used 2, 3 and 4% NaCl in TH and TCBS agar to examine blood from victims who had drowned in seawater (n=8) and in fresh water (n=7), as well as from victims who died near aquatic environments (non drowned; dry-land control, n=7). Bioluminescent colonies were detectable on 2-4% NaCl TH agar only from two victims that drowned in seawater. Bioluminescent colonies did not grow on TCBS agar. Blue colonies from all cadavers that had drowned in seawater (8/8) and in four of those that had drowned in fresh water (4/7) proliferated on TH agar containing 2% and/or 3% NaCl, but at 4% NaCl such colonies were detected only from cadavers that had drowned in seawater (8/8). Colonies from only one cadaver from seawater grew on TCBS agar. Furthermore, neither bioluminescent nor blue colonies were detected on TH agar containing 4% NaCl in samples from two cadavers found in an estuary (brackish water) who were thought to have been carried from areas of fresh water. Homologous analyses of the 16S rRNA gene revealed that the dominant colonies on TH agar containing 4% NaCl were marine bacteria (Photobacterium, Vibrio, Shewanella, Psychrobacter). Thus, proliferating bioluminescent and/or blue colonies detected in the blood of immersed cadavers using 4% NaCl TH agar, could help to establish drowning in seawater.

Topics: Agar; Bacteriological Techniques; Blood; Cadaver; Colony Count, Microbial; Drowning; Forensic Pathology; Fresh Water; Gammaproteobacteria; Humans; RNA, Bacterial; RNA, Ribosomal, 16S; Seawater; Sodium Chloride

Tissue Services (within NHS Blood and Transplant) plans to bring deceased donors to its state of the art retrieval suite at its new centre in Speke, Liverpool in air-conditioned transport at circa 20 degrees C but without dedicated active cooling. The aim of this study was to determine how quickly a refrigerated body would warm at different ambient temperatures using a gel-filled model. Two models of a human body were prepared consisting of neoprene wetsuits filled with approximately 7 or 18 l of a viscous solution, which once set has similar properties to ballistics gel. This gel consisted of 47.5% distilled water, 47.5% glycerol and 5% agar. Final "dummy" weights were 7.4 and 18.6 kg respectively, representing "virtual" weights of approximately 40 kg and 70 kg. A K-class thermocouple probe was then inserted into a "rectal" position within each model and the models were cooled to a series of different core temperatures: 5 degrees C, 10 degrees C and 15 degrees C and then were placed in an orbital incubator set at 20 degrees C or 30 degrees C ambient temperature. The rate of temperature increase, in the dummy, was measured, until the model's core temperature was close to the ambient temperature. This was done in triplicate for each size model and ambient temperature. Data indicate that increase in core temperature depends on the size of the model and the initial core temperature. For an equivalent donor weight of 70 kg and background temperature of 20 degrees C, core temperature rises from 5 degrees C to 9.2 degrees C; 10 degrees C to 13.3 degrees C and 15 degrees C to 15.5 degrees C after 2 h. The final core temperatures after 2 h are likely to retard bacterial growth, movement or contamination during transport. Cooling rate data indicated that a 70 kg donor equivalent cooled from 37 degrees C to 15 degrees C within 6 h in a cold room at 4 degrees C. This work has shown that a body can be transported without refrigeration and not cause further tissue deterioration as a result.

Topics: Agar; Body Temperature; Cadaver; Cold Temperature; Gels; Glycerol; Heating; Humans; Models, Anatomic; Water

Topics: Agar; Angiography; Arteries; Cadaver; Contrast Media; Ethylene Glycols; Histological Techniques; Humans; Methods; Staining and Labeling

Topics: Agar; Blood; Cadaver; Culture Media; Humans