北京中心城区硅藻分布的调查研究及破机罐消化法在溺死鉴定中的应用
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摘要
[背景]
溺死(death by drowning或drowning)是机械性窒息死亡的一种,溺死的法医学鉴定一直都是科研、检案的重点、难点问题。在法医实践中,诊断溺死的生物学和鉴证化学指标至今没有得到科学的证实与接受。当前,硅藻检验仍然是溺死诊断的“金标准(golden standard)"。在实验室检验硅藻的多种方法中,较为成熟且使用广泛的方法是硝酸乙醇消化法,但该法也有其自身的局限性。近年来新兴的破机罐消化法,对传统的硝酸乙醇消化法进行了改良,提高了实验操作的安全性,明确界定了实验用时、检材和试剂的消耗量,为法医学溺死鉴定的定性和定量检测带来了新的契机。此外,由于各水域环境复杂迥异,而硅藻种类繁多、形态迥异,不同水源的硅藻种属构成亦不相同,因此可以通过将检测出的硅藻种属及构成比,与水域中的硅藻进行对比,来推断死者的落水位置,从而为检案提供帮助。但迄今为止,无论是法医学领域还是环境生态学领域,都未见对北京地区硅藻种属分布进行研究的文献报道。
[目的]
1.研究北京市中心城区(西城区、东城区、朝阳区、海淀区、丰台区、石景山区)的主要河流及湖泊所含硅藻的种属分布；
2.从法医学角度,对北京、贵州省贵阳市、山东省滨州市、上海长江口海域、汉江流域(武汉段)、新疆乌伦古河流域、山西省娘子关泉域的硅藻优势种属进行横向比对；
3.鉴定并详细描述北京中心城区各水域所含硅藻的形态构造、生态习性；
4.摸索应用破机罐消化法检测家兔心脏、肺、肝、肾四种器官硅藻含量的最优实验条件(硝酸用量、消化温度、消化时间)；
5.比较破机罐消化法与硝酸乙醇消化法的差异。
[方法]
1.在北京市西城区、东城区、朝阳区、海淀区、丰台区、石景山区的主要河流及湖泊设置16个采水点,共采集80份水样；取水样置于破机罐中,加浓硝酸后放入鼓风干燥箱100℃消化0.5h,将罐中液体离心、洗涤、涂片；利用全景智能化扫描数字切片工作站(OLYMPUS-DotSlide)记取水样涂片中的硅藻数量,采集图像,对硅藻种属进行鉴别,统计分析各水域硅藻构成比,绘制硅藻分布图,比对北京、贵州省贵阳市、山东省滨州市、上海长江口海域、汉江流域(武汉段)、新疆乌伦古河流域、山西省娘子关泉域的硅藻优势种属差异。
2.空气栓塞处死健康家兔,取家兔的心脏组织,均分为180份,10g/份。将10g心脏组织剪碎,放入破机罐中,再向其中加入2mL梅尼小环藻藻种液待检。将180份待检心脏组织依据不同消化温度(100℃、105℃、110℃),不同硝酸用量(5mL10mL、15mL、20mL),不同消化时间(60min、75min、90min、105min、120min)随机组合分组。肺、肝、肾组织取材及实验分组同心脏。
3.按照实验设置消化组织,消化结束后,称重剩余液体残渣质量；液体离心、洗涤、涂片；筛选出完全消化组(肉眼观液体澄清透明,称量残渣质量<50mg)；对完全消化组定量检测,统计不同条件下的完整硅藻数量、硅藻碎片数量、硅藻总数、硅藻形态破坏程度(硅藻碎片／硅藻总数)。
4.在玻璃容器中盛满混有梅尼小环藻藻种液的蒸馏水,将家兔置于兔笼中,浸入水下40s,提出水面40s后重新浸入水下,重复5次后,彻底溺死家兔；取家兔的心脏、肺、肝、肾组织,每只家兔的脏器均分为A、B两组待检。
5.A组按照最佳实验条件设置行破机罐消化法,B组行硝酸乙醇消化法检测硅藻；称重两组剩余液体残渣质量：液体离心、洗涤、涂片；统计各组完整硅藻数量、硅藻碎片数量、硅藻总数、硅藻形态破坏程度(硅藻碎片／硅藻总数)。
6.所有实验数据采用SPSS17.0统计软件进行统计学分析。
[结果]
1.北京市西城区、东城区、朝阳区、海淀区、丰台区、石景山区主要河流及湖泊共检见10种硅藻藻属,分别隶属2纲5目10属,各水域所含硅藻种类及硅藻相对丰富度有所差异,部分河流可观察到显著的优势藻属；绘制了北京中心城区硅藻分布图；对不同地区(北京、贵州省贵阳市、山东省滨州市、上海长江口海域、汉江流域武汉段、新疆乌伦古河流域、山西省娘子关泉域)的硅藻优势种属进行横向比对,证实不同地区的硅藻分布差异显著,认为气候、地势、水体、周围工业化及社会化程度不同均会影响硅藻种属的构成。
2.心脏组织硅藻检验的最佳实验条件为：消化温度110℃,消化时间90min,组织(g)与硝酸用量(mL)的比例为2：3；肺组织硅藻检验的最佳实验条件为：消化温度100℃,消化时间105min,组织(g)与硝酸用量(mL)的比例为2：3；肝组织硅藻检验的最佳实验条件为：消化温度105℃,消化时间105min,组织(g)与硝酸用量(mL)的比例为2：3；肾组织硅藻检验的最佳实验条件为：消化温度110℃,消化时间120min,组织(g)与硝酸用量(mL)的比例为2：3。
3.破机罐消化法组织残渣能够达到完全消化的客观标准,消化程度稳定；硝酸乙醇消化法消化程度不稳定,无法达到完全消化的客观标准；破机罐消化法的硅藻检出率高于硝酸乙醇消化法,其硅藻破坏率低于硝酸乙醇消化法。
[结论]
1.首次将全景智能化扫描数字切片的理论和技术应用于法医学硅藻鉴定,证实了利用该技术鉴定硅藻的可行性。
2.总结了北京市中心城区各水域中硅藻分布的特点,首次建立了北京中心城区水域的硅藻形态及相对构成比数据库,并绘制了相应的北京城区硅藻分布图,为北京地区溺死案件中相应水域的硅藻比对提供了基础检测平台。
3.首次从法医学角度,将不同地区(北京、贵州省贵阳市、山东省滨州市、上海长江口海域、汉江流域武汉段、新疆乌伦古河、山西省娘子关泉域)的硅藻优势种属进行横向比对,进一步证实了硅藻检验在刑事侦查中用以推断死者落水位置的重大意义。
4.鉴定并详细描述了北京中心城区河流及湖泊所含硅藻的形态构造、生态习性,为法医工作者依据硅藻形态比对判断水中尸体落水位置提供帮助。
5.首次证实了机体心、肺、肝、肾组织由于质实程度不同,应用的破机罐消化法需要的硝酸用量、消化温度及消化时间各不相同,每种器官在最优实验条件设置下,能够获得硅藻的高检出率及低形态破坏率的最佳契合点。
6.与硝酸乙醇消化法比较,破机罐消化法具有以下几个优点：①硝酸用量少；②环境污染小；③有机组织消解完全,消化程度具有稳定性；④硅藻破坏程度小；⑤硅藻检出率高；⑥硅藻定性定量分析的准确度高。
[Backgound]
Drowning is defined as death from asphyxia within24hours of submersion in water, and always a problem in judicial practice and scientific research in forensic pathology. It has not been accepted and unified that some biological and chemical indexes were sufficiently specific or sensitive for diagnosis of drowning. At present, the detection of foreign plant elements such as diatoms is assessed and the value of diatom test as the "golden standard" is evaluated. In addition, it enables an estimate of position falled in water by testing the diatom species and proportions in bodies and comparing to which are observed among the different waters, because of various shapes, structures, taxonomic characters of each diatom and different features, species compositions of distribution of diatoms in different regions. So the research on diatom test has important value for the key evidence in case of drowning. Nitric acid added ethanol method is the most well rounded and widely used method at present, despite its limitations, among the many diatom laboratory examination methods. Disorganization can method is a new technology for testing diatom developed in recent years, with specific mensurable criteria of time consuming, amout of samples and volume of reagents. It was modified on nitric acid added ethanol method, and had been providing new opportunities in forensic expertise.
[Objectives]
1. Study the quantity and distribution of diatoms in Xicheng, Dongcheng, Chaoyang, Haidian, Fengtai, Shijingshan District of Beijing City, and draw the map of distribution of diatoms species in central city of Beijing;
2. Contrast the dominant population of diatoms about Beijing, Guiyang Municipality, Binzhou, the Changjiang River estuary, Hanjiang catchment, Ulungur River Valley in Xinjiang, Niangziguan Karst spring.
3. Present a review on species composition, shapes, structures, taxonomic characters for each taxon, and describes the distribution of diatoms on central city of Beijing;
4. To study the optimal experimental conditions (nitric acid consumption, temperature and time for complete digestion) to detect ratio of diatom of rabbit heart, lungs, liver and kidneys by using disorganization can;
5. To compare the difference between disorganization can and nitric acid added ethanol method.
[Methods]
1.80water samples collected from16waters of central city of Beijing were examined by the method of disorganization can, which digested by nitric acid at100℃for0.5h, then centrifuging, washing and conventional smear. To identify the diatom species and proportions of water samples, collect image data under the DotSlide microscope, and draw the map of distribution of diatoms species in central city of Beijing. Moreover, to contrast the dominant population of diatoms about Beijing, Guiyang Municipality, Binzhou, the Changjiang River estuary, Hanjiang catchment, Ulungur River Valley in Xinjiang, Niangziguan Karst spring.
2. Rabbits were put to death by air embolism. Four kinds of rabbit tissue of heart, lung, liver and kidney were sampled, and each kind of tissue was divided into180parts, which put in disorganization cans with diatoms for subsequent test. All the cans were detected in groups randomly according to3indexes of experimental condition, including nitric acid consumption (5mL、10mL、15mL、20mL), temperature (100℃、105℃、110℃) and time (60min、75min、90min、105min、120min) for complete digestion.
3. Four kinds of rabbit tissue of heart, lung, liver and kidney were digested by disorganization can with different experimental conditions, respectively. All the groups weighted the surplus of organs, selected the groups digested completely (the surplus was less than50mg), and recorded the clarity of background. For the groups digested completely, the statistical quantity of unbroken diatoms, diatom debris, and demolition rate were counted.
4. Rabbits were put to death by drowning in a large, glass container with distilled water, mixed diatoms. Four kinds of rabbit tissue of heart, lung, liver and kidney were sampled, and tissues of every rabbit were divided into two groups randomly.
5. Four kinds of rabbit tissue of heart, lung, liver and kidney were completely digested by disorganization can with optimal experimental conditions for group A and nitric acid added ethanol method for group B, respectively. Both groups weighted the surplus of organs, and counted the statistical quantity of unbroken diatoms, diatom debris, and demolition rate.
6. The data obtained were statistical analyzed by SPSS17.0software.
[Results]
1. Having analyzed80water samples collected from16waters of Xicheng, Dongcheng, Chaoyang, Haidian, Fengtai, Shijingshan District of Beijing in September and October2011, and mapped the distribution of diatoms species in central city of Beijing, a total of10taxa belonging to10genera,5orders,2classes were identified. Significant differences of diatom species and proportions were observed among the different waters. After contrasting the dominant population of diatoms about Beijing, Guiyang Municipality, Binzhou, the Changjiang River estuary, Hanjiang catchment, Ulungur River Valley in Xinjiang, Niangziguan Karst spring, it is confirmed the distribution of diatoms species was affected by the differences of climate, terrain, water.
2. The most appropriate conditions for diatom test in heart tissue were digested at110℃, lasted90min, and the tissues (g) and nitric acid (mL) was at the ratio of2:3. The most appropriate conditions for diatom test in lung tissue were digested at100℃, lasted105min, and the tissues (g) and nitric acid (mL) was at the ratio of2:3. The most appropriate conditions for diatom test in liver tissue were digested at105℃, lasted105min, and the tissues (g) and nitric acid (mL) was at the ratio of2:3. The most appropriate conditions for diatom test in renal tissue were digested at110℃, lasted120min, and the tissues (g) and nitric acid (mL) was at the ratio of2:3.
3. The surplus of organs came up to the standard of complete digestion, and degree of digestion had greater stability by using disorganization can, but nitric acid added ethanol method was not. Moreover, disorganization can had preferable detection rate and lower demolition rate compare to nitric acid added ethanol method.
[Conclusion]
1. This study confirmed the feasibility of using DotSlide microscope.
2. The features of distribution of diatoms in central city of Beijing were summarized, and the map of distribution of diatoms species in central city of Beijing was drew for the first time. We also discussed its meaning in criminal detection. Furthermore, these theoretical and experimental investigations have laid a solid foundation for establishing the diatom database in central city of Beijing.
3. This strdy first contrasted the dominant population of diatoms about Beijing, Guiyang Municipality, Binzhou, the Changjiang River estuary, Hanjiang catchment, Ulungur River Valley in Xinjiang, Niangziguan Karst spring, it is confirmed the value of examination of diatom in forensic science.
4. This paper presents a review on species composition, shapes, structures, taxonomic characters for each taxon, and describes the distribution of diatoms on central city of Beijing. And the study has laid a solid foundation for determining the position of drowning based on the structures of diatoms.
5. The optimal experimental conditions (nitric acid consumption, temperature and time for complete digestion) varied in different tissues because of the quality of heart, lungs, liver and kidneys, which ensured preferable detection rate.
6. In contrast with nitric acid added ethanol method, the features of disorganization can are shown as follows:①Less amount of nitric acid;②Little environmental pollution;③Digestion is complete and stabilized;④Diatom damage small;⑤High detection rate;⑥High accuracy of diatom qualitative/quantitative analysis.
溺死(death by drowning或drowning)是机械性窒息死亡的一种,溺死的法医学鉴定一直都是科研、检案的重点、难点问题。在法医实践中,诊断溺死的生物学和鉴证化学指标至今没有得到科学的证实与接受。当前,硅藻检验仍然是溺死诊断的“金标准(golden standard)"。在实验室检验硅藻的多种方法中,较为成熟且使用广泛的方法是硝酸乙醇消化法,但该法也有其自身的局限性。近年来新兴的破机罐消化法,对传统的硝酸乙醇消化法进行了改良,提高了实验操作的安全性,明确界定了实验用时、检材和试剂的消耗量,为法医学溺死鉴定的定性和定量检测带来了新的契机。此外,由于各水域环境复杂迥异,而硅藻种类繁多、形态迥异,不同水源的硅藻种属构成亦不相同,因此可以通过将检测出的硅藻种属及构成比,与水域中的硅藻进行对比,来推断死者的落水位置,从而为检案提供帮助。但迄今为止,无论是法医学领域还是环境生态学领域,都未见对北京地区硅藻种属分布进行研究的文献报道。
[目的]
1.研究北京市中心城区(西城区、东城区、朝阳区、海淀区、丰台区、石景山区)的主要河流及湖泊所含硅藻的种属分布；
2.从法医学角度,对北京、贵州省贵阳市、山东省滨州市、上海长江口海域、汉江流域(武汉段)、新疆乌伦古河流域、山西省娘子关泉域的硅藻优势种属进行横向比对；
3.鉴定并详细描述北京中心城区各水域所含硅藻的形态构造、生态习性；
4.摸索应用破机罐消化法检测家兔心脏、肺、肝、肾四种器官硅藻含量的最优实验条件(硝酸用量、消化温度、消化时间)；
5.比较破机罐消化法与硝酸乙醇消化法的差异。
[方法]
1.在北京市西城区、东城区、朝阳区、海淀区、丰台区、石景山区的主要河流及湖泊设置16个采水点,共采集80份水样；取水样置于破机罐中,加浓硝酸后放入鼓风干燥箱100℃消化0.5h,将罐中液体离心、洗涤、涂片；利用全景智能化扫描数字切片工作站(OLYMPUS-DotSlide)记取水样涂片中的硅藻数量,采集图像,对硅藻种属进行鉴别,统计分析各水域硅藻构成比,绘制硅藻分布图,比对北京、贵州省贵阳市、山东省滨州市、上海长江口海域、汉江流域(武汉段)、新疆乌伦古河流域、山西省娘子关泉域的硅藻优势种属差异。
2.空气栓塞处死健康家兔,取家兔的心脏组织,均分为180份,10g/份。将10g心脏组织剪碎,放入破机罐中,再向其中加入2mL梅尼小环藻藻种液待检。将180份待检心脏组织依据不同消化温度(100℃、105℃、110℃),不同硝酸用量(5mL10mL、15mL、20mL),不同消化时间(60min、75min、90min、105min、120min)随机组合分组。肺、肝、肾组织取材及实验分组同心脏。
3.按照实验设置消化组织,消化结束后,称重剩余液体残渣质量；液体离心、洗涤、涂片；筛选出完全消化组(肉眼观液体澄清透明,称量残渣质量<50mg)；对完全消化组定量检测,统计不同条件下的完整硅藻数量、硅藻碎片数量、硅藻总数、硅藻形态破坏程度(硅藻碎片／硅藻总数)。
4.在玻璃容器中盛满混有梅尼小环藻藻种液的蒸馏水,将家兔置于兔笼中,浸入水下40s,提出水面40s后重新浸入水下,重复5次后,彻底溺死家兔；取家兔的心脏、肺、肝、肾组织,每只家兔的脏器均分为A、B两组待检。
5.A组按照最佳实验条件设置行破机罐消化法,B组行硝酸乙醇消化法检测硅藻；称重两组剩余液体残渣质量：液体离心、洗涤、涂片；统计各组完整硅藻数量、硅藻碎片数量、硅藻总数、硅藻形态破坏程度(硅藻碎片／硅藻总数)。
6.所有实验数据采用SPSS17.0统计软件进行统计学分析。
[结果]
1.北京市西城区、东城区、朝阳区、海淀区、丰台区、石景山区主要河流及湖泊共检见10种硅藻藻属,分别隶属2纲5目10属,各水域所含硅藻种类及硅藻相对丰富度有所差异,部分河流可观察到显著的优势藻属；绘制了北京中心城区硅藻分布图；对不同地区(北京、贵州省贵阳市、山东省滨州市、上海长江口海域、汉江流域武汉段、新疆乌伦古河流域、山西省娘子关泉域)的硅藻优势种属进行横向比对,证实不同地区的硅藻分布差异显著,认为气候、地势、水体、周围工业化及社会化程度不同均会影响硅藻种属的构成。
2.心脏组织硅藻检验的最佳实验条件为：消化温度110℃,消化时间90min,组织(g)与硝酸用量(mL)的比例为2：3；肺组织硅藻检验的最佳实验条件为：消化温度100℃,消化时间105min,组织(g)与硝酸用量(mL)的比例为2：3；肝组织硅藻检验的最佳实验条件为：消化温度105℃,消化时间105min,组织(g)与硝酸用量(mL)的比例为2：3；肾组织硅藻检验的最佳实验条件为：消化温度110℃,消化时间120min,组织(g)与硝酸用量(mL)的比例为2：3。
3.破机罐消化法组织残渣能够达到完全消化的客观标准,消化程度稳定；硝酸乙醇消化法消化程度不稳定,无法达到完全消化的客观标准；破机罐消化法的硅藻检出率高于硝酸乙醇消化法,其硅藻破坏率低于硝酸乙醇消化法。
[结论]
1.首次将全景智能化扫描数字切片的理论和技术应用于法医学硅藻鉴定,证实了利用该技术鉴定硅藻的可行性。
2.总结了北京市中心城区各水域中硅藻分布的特点,首次建立了北京中心城区水域的硅藻形态及相对构成比数据库,并绘制了相应的北京城区硅藻分布图,为北京地区溺死案件中相应水域的硅藻比对提供了基础检测平台。
3.首次从法医学角度,将不同地区(北京、贵州省贵阳市、山东省滨州市、上海长江口海域、汉江流域武汉段、新疆乌伦古河、山西省娘子关泉域)的硅藻优势种属进行横向比对,进一步证实了硅藻检验在刑事侦查中用以推断死者落水位置的重大意义。
4.鉴定并详细描述了北京中心城区河流及湖泊所含硅藻的形态构造、生态习性,为法医工作者依据硅藻形态比对判断水中尸体落水位置提供帮助。
5.首次证实了机体心、肺、肝、肾组织由于质实程度不同,应用的破机罐消化法需要的硝酸用量、消化温度及消化时间各不相同,每种器官在最优实验条件设置下,能够获得硅藻的高检出率及低形态破坏率的最佳契合点。
6.与硝酸乙醇消化法比较,破机罐消化法具有以下几个优点：①硝酸用量少；②环境污染小；③有机组织消解完全,消化程度具有稳定性；④硅藻破坏程度小；⑤硅藻检出率高；⑥硅藻定性定量分析的准确度高。
[Backgound]
Drowning is defined as death from asphyxia within24hours of submersion in water, and always a problem in judicial practice and scientific research in forensic pathology. It has not been accepted and unified that some biological and chemical indexes were sufficiently specific or sensitive for diagnosis of drowning. At present, the detection of foreign plant elements such as diatoms is assessed and the value of diatom test as the "golden standard" is evaluated. In addition, it enables an estimate of position falled in water by testing the diatom species and proportions in bodies and comparing to which are observed among the different waters, because of various shapes, structures, taxonomic characters of each diatom and different features, species compositions of distribution of diatoms in different regions. So the research on diatom test has important value for the key evidence in case of drowning. Nitric acid added ethanol method is the most well rounded and widely used method at present, despite its limitations, among the many diatom laboratory examination methods. Disorganization can method is a new technology for testing diatom developed in recent years, with specific mensurable criteria of time consuming, amout of samples and volume of reagents. It was modified on nitric acid added ethanol method, and had been providing new opportunities in forensic expertise.
[Objectives]
1. Study the quantity and distribution of diatoms in Xicheng, Dongcheng, Chaoyang, Haidian, Fengtai, Shijingshan District of Beijing City, and draw the map of distribution of diatoms species in central city of Beijing;
2. Contrast the dominant population of diatoms about Beijing, Guiyang Municipality, Binzhou, the Changjiang River estuary, Hanjiang catchment, Ulungur River Valley in Xinjiang, Niangziguan Karst spring.
3. Present a review on species composition, shapes, structures, taxonomic characters for each taxon, and describes the distribution of diatoms on central city of Beijing;
4. To study the optimal experimental conditions (nitric acid consumption, temperature and time for complete digestion) to detect ratio of diatom of rabbit heart, lungs, liver and kidneys by using disorganization can;
5. To compare the difference between disorganization can and nitric acid added ethanol method.
[Methods]
1.80water samples collected from16waters of central city of Beijing were examined by the method of disorganization can, which digested by nitric acid at100℃for0.5h, then centrifuging, washing and conventional smear. To identify the diatom species and proportions of water samples, collect image data under the DotSlide microscope, and draw the map of distribution of diatoms species in central city of Beijing. Moreover, to contrast the dominant population of diatoms about Beijing, Guiyang Municipality, Binzhou, the Changjiang River estuary, Hanjiang catchment, Ulungur River Valley in Xinjiang, Niangziguan Karst spring.
2. Rabbits were put to death by air embolism. Four kinds of rabbit tissue of heart, lung, liver and kidney were sampled, and each kind of tissue was divided into180parts, which put in disorganization cans with diatoms for subsequent test. All the cans were detected in groups randomly according to3indexes of experimental condition, including nitric acid consumption (5mL、10mL、15mL、20mL), temperature (100℃、105℃、110℃) and time (60min、75min、90min、105min、120min) for complete digestion.
3. Four kinds of rabbit tissue of heart, lung, liver and kidney were digested by disorganization can with different experimental conditions, respectively. All the groups weighted the surplus of organs, selected the groups digested completely (the surplus was less than50mg), and recorded the clarity of background. For the groups digested completely, the statistical quantity of unbroken diatoms, diatom debris, and demolition rate were counted.
4. Rabbits were put to death by drowning in a large, glass container with distilled water, mixed diatoms. Four kinds of rabbit tissue of heart, lung, liver and kidney were sampled, and tissues of every rabbit were divided into two groups randomly.
5. Four kinds of rabbit tissue of heart, lung, liver and kidney were completely digested by disorganization can with optimal experimental conditions for group A and nitric acid added ethanol method for group B, respectively. Both groups weighted the surplus of organs, and counted the statistical quantity of unbroken diatoms, diatom debris, and demolition rate.
6. The data obtained were statistical analyzed by SPSS17.0software.
[Results]
1. Having analyzed80water samples collected from16waters of Xicheng, Dongcheng, Chaoyang, Haidian, Fengtai, Shijingshan District of Beijing in September and October2011, and mapped the distribution of diatoms species in central city of Beijing, a total of10taxa belonging to10genera,5orders,2classes were identified. Significant differences of diatom species and proportions were observed among the different waters. After contrasting the dominant population of diatoms about Beijing, Guiyang Municipality, Binzhou, the Changjiang River estuary, Hanjiang catchment, Ulungur River Valley in Xinjiang, Niangziguan Karst spring, it is confirmed the distribution of diatoms species was affected by the differences of climate, terrain, water.
2. The most appropriate conditions for diatom test in heart tissue were digested at110℃, lasted90min, and the tissues (g) and nitric acid (mL) was at the ratio of2:3. The most appropriate conditions for diatom test in lung tissue were digested at100℃, lasted105min, and the tissues (g) and nitric acid (mL) was at the ratio of2:3. The most appropriate conditions for diatom test in liver tissue were digested at105℃, lasted105min, and the tissues (g) and nitric acid (mL) was at the ratio of2:3. The most appropriate conditions for diatom test in renal tissue were digested at110℃, lasted120min, and the tissues (g) and nitric acid (mL) was at the ratio of2:3.
3. The surplus of organs came up to the standard of complete digestion, and degree of digestion had greater stability by using disorganization can, but nitric acid added ethanol method was not. Moreover, disorganization can had preferable detection rate and lower demolition rate compare to nitric acid added ethanol method.
[Conclusion]
1. This study confirmed the feasibility of using DotSlide microscope.
2. The features of distribution of diatoms in central city of Beijing were summarized, and the map of distribution of diatoms species in central city of Beijing was drew for the first time. We also discussed its meaning in criminal detection. Furthermore, these theoretical and experimental investigations have laid a solid foundation for establishing the diatom database in central city of Beijing.
3. This strdy first contrasted the dominant population of diatoms about Beijing, Guiyang Municipality, Binzhou, the Changjiang River estuary, Hanjiang catchment, Ulungur River Valley in Xinjiang, Niangziguan Karst spring, it is confirmed the value of examination of diatom in forensic science.
4. This paper presents a review on species composition, shapes, structures, taxonomic characters for each taxon, and describes the distribution of diatoms on central city of Beijing. And the study has laid a solid foundation for determining the position of drowning based on the structures of diatoms.
5. The optimal experimental conditions (nitric acid consumption, temperature and time for complete digestion) varied in different tissues because of the quality of heart, lungs, liver and kidneys, which ensured preferable detection rate.
6. In contrast with nitric acid added ethanol method, the features of disorganization can are shown as follows:①Less amount of nitric acid;②Little environmental pollution;③Digestion is complete and stabilized;④Diatom damage small;⑤High detection rate;⑥High accuracy of diatom qualitative/quantitative analysis.
引文
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