Comparison of Application of MD-VF-Auto SEM Method and Plankton Gene Multiplex PCR System in the Diagnosis of Drowning / 法医学杂志

OBJECTIVES@#To compare the application effect of microwave digestion - vacuum filtration - automated scanning electron microscopy (MD-VF-Auto SEM) method and plankton gene multiplex PCR system in the diagnosis of drowning.@*METHODS@#Lung, liver and kidney tissue of 10 non-drowning cases and 50 drowning cases were prepared for further MD-VF-Auto SEM method analysis and plankton gene multiplex PCR system analysis. The positive detection rate of the two methods in each tissue was calculated.@*RESULTS@#The positive rate of the MD-VF-Auto SEM method detecting diatoms in drowning cases was 100%, and few diatoms were detected in the liver and kidney tissues of 6 non-drowning cases. By using the plankton gene multiplex PCR system, the diatom positive rate of drowning cases was 84%, and all the non-drowning cases were negative. There were significant differences in the positive rate of the liver, kidney tissues between MD-VF-Auto SEM method and plankton gene multiplex PCR system (P<0.05), as well as the total positive rate of cases. However, no significant differences were found in the positive rates of lung tissues (P>0.05).@*CONCLUSIONS@#MD-VF-Auto SEM method is more sensitive than plankton gene multiplex PCR system in diatom test. But the plankton gene multiplex PCR system can also detect plankton other than diatoms. Combination of the two methods can provide a more reliable basis for the diagnosis of drowning.

Application of Chlorophyte ChlB Gene and Cyanophyte NIES Gene in the Detection of Drowning-Related Plankton / 法医学杂志

Objective To construct a polymerase chain reaction-capillary electrophoresis （PCR-CE） detection method using ChlB gene and NIES gene, investigate the method's specificity and sensitivity, and to evaluate its application value in drowning diagnosis. Methods The specific primers ChlB and NIES were designed for the conserved sequence of chlorophyte ChlB gene and cyanophyte NIES gene in GenBank to construct PCR-CE detection method; 50 species of standard DNA samples were amplified; the sensitivity was determined by gradient concentration detection of positive standard samples; 25 actual cadaver lung tissue samples （drowned： 20, natural death： 5） were detected, and the simultaneous detection results of microwave digestion-vacuum filtration-automated scanning electron microscopy （MD-VF-Auto SEM） were simultaneously compared. Results The minimum DNA detection concentration of primers ChlB and NIES was 0.161 ng and 0.109 ng, respectively, which could specifically amplify chlorophyte （Chlorella pyrenoidosa） and cyanophyte ［Microcystis aeruginosa （producing and not producing toxin）］ widespread in water. The product fragments were 156 bp and 182 bp, respectively. The results of non-drowning tissues were negative. Conclusion This method has high sensitivity and specificity. It can be applied to the detection of plankton related to drowning and combined with MD-VF-Auto SEM method, can increase the detection range of plankton related to drowning and improve the evidence power of drowning diagnosis.

A molecular method for the detection of sally lightfoot crab larvae (Grapsus grapsus, Brachyura, Grapsidae) in plankton samples

The decapod Grapsus grapsus is commonly found on oceanic islands of the Pacific and Atlantic coasts of the Americas. In this study, a simple, quick and reliable method for detecting its larvae in plankton samples is described, which makes it ideal for large-scale studies of larval dispersal patterns in the species.

Estudio comparativo de la estructura del bacterioplancton en aguas del Mar Argentino mediante el método de pirosecuenciación 454 tag / A comparative study of bacterioplankton structure in Argentinian Sea waters by the 454 - tag pyrosequencing method

El presente estudio brinda la primera información sobre diversidad y abundancia de las comunidades microbianas en dos ambientes del Mar Argentino obtenida mediante la técnica de pirosecuenciación tag ribosomal 454. Dentro del dominio Bacteria, se observaron más de 4 600 secuencias únicas a partir de 36 188 amplicones de tags y se identificaron 280 filotipos. Además, se detectaron cerca de 2 700 secuencias únicas a partir de más de 47 700 tags pertenecientes al dominio Archaea, lo que definió sólo 5 filotipos diferentes. La distancia de Jaccard presentó valores de 0,6 para bacterias y de 0,2 para arqueas, esto indica mayor diferencia entre las bacterias en los dos sitios. En el ambiente marino los filotipos más dominantes fueron Bacteroidetes Flavobacteriaceae, Proteobacteria Gammaproteobacteria, Proteobacteria Rhodobacteraceae y Proteobacteria Rickettsiales SAR11, mientras que en el estuario predominaron Pseudoalteromonadaceae Pseudoalteromonas, Proteobacteria Gammaproteobacteria, Proteobacteria Shewanella y Proteobacteria Rickettsiales SAR11. Los 2 filotipos de arqueas encontrados en mayor proporción fueron Archaea Euryarchaeota y Archaea Crenarchaeota. Las secuencias tag más numerosas representaron taxa caracterizados previamente, aunque también se halló un elevado número de filotipos de gran diversidad y de baja abundancia, que forman parte de la denominada "biosfera rara", aún no explorada, que pueden tener un papel ecológico crucial.

The present study provides the first information about diversity and abundance of microbial communities in two environments of the Argentinian Sea by the 454 - tag pyrosequencing technique. We observed more than 4,600 unique bacterial sequences from 36,188 tag amplicons, forming 280 phylotypes. In addition, nearly 2,700 unique sequences from more than 47,700 tags identified as Archaea, defined only 5 different phylotypes. The Jaccard distance (0.6 for Bacteria and 0.2 for Archaea) indicated higher differences among Bacteria rather than among Archaea in both studied sites. The dominant phylotypes in marine environment were Bacteroidetes Flavobacteriaceae, Proteobacteria Gammaproteobacteria, Proteobacteria Rhodobacteraceae and Proteobacteria Rickettsiales SAR11; and Pseudoalteromonadaceae Pseudoalteromonas, Proteobacteria Gammaproteobacteria, Proteobacteria Shewanella, Proteobacteria Rickettsiales SAR11 in the estuary sampling site. Archaea Euryarchaeota and Archaea Crenarchaeota were the major archaeal phylotypes found. The most abundant tag sequences included previously characterized taxa, although we also retrieved a large number of highly diverse, low-abundant phylotypes which constitute a largely unexplored "rare" biosphere. These microorganisms could have a crucial ecological role.