Arsenic speciation and valence / 中国中药杂志

As arsenic widely exists in nature and has been used in the pharmaceutical preparations, the traditional Chinese medicine(TCM) with arsenic include realgar(As_2S_2 or As_4S_4), orpiment(As_2S_3), and white arsenic(As_2O_3). Among the above representative medicine, the TCM compound formulas with realgar are utilized extensively. Just in Chinese Pharmacopoeia(2020 edition), there are 37 Chinese patent medicines including realgar. The traditional element analysis focuses on the detection of the total amount of elements, which neglects the study on the speciation and valence of elements. The activity, toxicity, bioavailability, and metabolic pathways of arsenic in vivo are closely related to the existence of its form, and different forms of arsenic have different effects on organisms. Therefore, the study on the speciation and valence of arsenic is of great importance for arsenic-containing TCMs and their compound formulas. This paper reviewed four aspects of the speciation and valence of arsenic, including property, absorption and metabolism, toxicity, and analytical assay.

SD bioline malaria rapid diagnostic test performance and time to become negative after treatment of malaria infection in southwest Nigerian children

Context and Aim: Given the challenges of microscopy, we compared its performance with SD Bioline malaria rapid diagnostic test (MRDT) and polymerase chain reaction (PCR) and evaluated the time it took for positive results to become negative after treatment of children with acute uncomplicated malaria. Subjects and Methods: We present the report of 485 participants with complete MRDT, microscopy, and PCR data out of 511 febrile children aged 3­59 months who participated in a cohort study over a 12 month period in rural and urban areas of Ibadan, Nigeria. MRDT positive children received antimalaria and tested at every visit over 28 days. Speciation was also carried out by PCR. Results: With microscopy as the gold standard, SD-Bioline™ had 95.2% sensitivity, 66.4% specificity, 67.5% positive predictive value (PPV), and 94.9 negative predictive value (NPV), while with PCR the findings were 84.3% sensitivity, 66.5% specificity, 72.7% PPV, and 80.1% NPV. PCR speciation of malaria parasites revealed 91.6% Plasmodium falciparum, 18.9% Plasmodium malariae, and 4.4% Plasmodium ovale. Among the 47 children with P. malariae infections, 66.0% were coinfected with P. falciparum, while 54.6% cases of P. ovale occurred as coinfections with P. falciparum. The median time to a negative MRDT was 23.2 days, while the median time to a negative malaria microscopy was 3.8 days. The two survival curves were significantly different. Conclusions: The SD BiolineTM MRDT performed well, with remarkable persistence of rapid test-positive for an average of 23 days post treatment. The prevalence of P. malaria is somewhat greater than expected.

Tin speciation in the blood plasma of workers occupationally exposed in a cassiterite ore processing industry / Especiação de estanho no plasma sanguíneo de trabalhadores expostos ocupacionalmente em uma indústria de processamento de minério de cassiterita

ABSTRACT Mining is a high-risk activity due to its dangerous processes. Tin (Sn) is obtained from cassiterite ore and mining activities expose workers to the metal. Chronic exposure to Sn may cause pneumoconiosis, gastrointestinal and hematological effects, among others. This work aimed to assess the exposure of workers to tin in a cassiterite ore processing industry, using the speciation analysis in blood plasma. Twelve subjects donated the blood samples; six were occupationally exposed to Sn. Size exclusion chromatography separated proteins in blood plasma; a graphite furnace atomic absorption spectrometer determined total tin in the plasma and eluted fractions, while SDS-PAGE determined molecular masses of proteins. Tin levels in the workers' plasma were four times higher than in the reference individuals. After fractionation, the metal only appeared in the total inclusion volume, not being possible to confirm the binding of tin to proteins, which certainly modifies their functions and impair workers' health. Despite that, the work process needs to change since Sn levels in the workers' plasma pointed to metal exposure. Further works are necessary to clarify whether the metal is free or bound to small proteins in blood plasma and understand the true impact of tin on workers' health.

RESUMO A mineração é uma atividade de alto risco devido aos seus processos perigosos. O estanho (Sn) é obtido do minério de cassiterita e as atividades da mineração expõem os trabalhadores ao metal. A exposição crônica ao Sn pode causar pneumoconiose, gastrointestinal e hematológica entre outros efeitos. Este trabalho avaliou a exposição de trabalhadores ao estanho em uma indústria de processamento de minério de cassiterita, utilizando a análise de especiação no plasma sanguíneo. Doze indivíduos doaram amostras de sangue, sendo seis expostos ocupacionalmente ao Sn. A SEC separou as proteínas do plasma sanguíneo, a GFAAS determinou a concentração total de estanho no plasma sanguíneo e frações eluídas, enquanto o SDS-PAGE determinou as massas moleculares das proteínas. O plasma dos trabalhadores apresentou níveis quatro vezes maiores do que os indivíduos de referência. Após fracionamento, Sn só apareceu no volume de inclusão, não sendo possível confirmar sua ligação às proteínas. Contudo, o processo de trabalho precisa mudar, pois os níveis de Sn no plasma dos trabalhadores apontam para exposição ao metal. Outros trabalhos são necessários para esclarecer se o metal está livre ou ligado a pequenas proteínas do plasma e entender o verdadeiro impacto do estanho na saúde dos trabalhadores.

DNA Barcoding unveils cryptic lineages of Hoplias malabaricus from Northeastern Brazil / DNA Barcoding revela linhagens crípticas de Hoplias malabaricus no nordeste do Brasil

Abstract The trahira or wolf fish - Hoplias malabaricus- is a valid species, although recent cytogenetic and molecular studies have indicated the existence of a species complex. In this context, the present study analyzed the mitochondrial COI marker to determine the levels of genetic diversity of specimens from the Brazilian state of Maranhão, and verify the occurrence of distinct lineages within the study area. Samples were collected from the basins of the Turiaçu, Pindaré, Mearim, Itapecuru, and Parnaíba rivers. A 630-bp fragment was obtained from 211 specimens, with 484 conserved and 108 variable sites, and 60 haplotypes (Hd = 0,947; π = 0,033). The phylogenetic analyses indicated the existence of three distinct lineages of H. malabaricus from Maranhão. Genetic distances of 1.5-8.2% were found between all the populations analyzed, while the variation between haplogroups ranged from 2.1% to 7.7%. The AMOVA indicated that most of the molecular variation was found among groups, with high FST values. The high levels of genetic variability found in the present study are supported by the available cytogenetic data. These findings reinforce the need for the development of effective programs of conservation and management independently for each river basin, in order to preserve the genetic variability found in this taxon.

Resumo A traíra - Hoplias malabaricus- é uma espécie válida, embora recentes estudos citogenéticos e moleculares tenham indicado a existência de um complexo de espécies. Neste contexto, o presente estudo analisou o marcador mitocondrial COI para determinar os níveis de diversidade genética dos espécimes do estado do Maranhão e verificar a ocorrência de linhagens distintas dentro da área de estudo. As amostras foram coletadas nas bacias dos rios Turiaçu, Pindaré, Mearim, Itapecuru e Parnaíba. As análises filogenéticas indicaram a existência de três linhagens distintas nas populações do Maranhão. Obteve-se um fragmento de 630 pb de 211 espécimes, com 484 sítios conservados, 108 variáveis e 60 haplótipos (Hd = 0,947; π = 0,033). As análises filogenéticas indicaram a ocorrência de três linhagens distintas de H. malabaricus do Maranhão. Distâncias genéticas de 1.5 a 8.2% foram encontradas entre todas as populações analisadas, enquanto a variação entre os haplogrupos variou de 2.1% a 7.7%. A AMOVA indicou que a maior variação molecular foi entre os grupos, com altos valores de FST. Os altos níveis de variabilidade genética encontrados no presente estudo são suportados pelos dados citogenéticos disponíveis. Essas descobertas reforçam a necessidade de desenvolver programas de conservação e manejo independentemente para cada bacia hidrográfica, a fim de preservar a variabilidade genética encontrada neste táxon.

Cryptic speciation in populations of the genus Aphyocharax (Characiformes: Characidae) from eastern Amazon coastal river drainages and surroundings revealed by single locus species delimitation methods

Recent studies in eastern Amazon coastal drainages and their surroundings have revealed new fish species that sometimes exhibit little morphological differentiation (cryptic species). Thus, we used a DNA-based species delimitation approach to test if populations showing the morphotype and typical character states of the Aphyocharax avary holotype correspond either to A. avary or A. brevicaudatus, two known species from the region, or if they form independent lineages, indicating cryptic speciation. WP and GMYC analyses recovered five lineages (species) in the ingroup, while a bPTP analysis delimited three lineages. ABGD analyses produced two possible results: one corroborating the WP and GMYC methods and another corroborating the bPTP method. All methods indicate undescribed cryptic species in the region and show variation from at least 1 to 4 species in the ingroup, depending on the approach, corroborating previous studies, and revealing this region as a possible hotspot for discovering undescribed fish species.(AU)

Estudos recentes nas drenagens costeiras da Amazônia oriental e seus arredores revelaram novas espécies de peixes que às vezes exibem pouca diferenciação morfológica (espécies crípticas). Assim, usamos uma abordagem de delimitação de espécies baseada em DNA para testar se as populações que apresentam o morfotipo e os estados de caráter típicos do holótipo Aphyocharax avary correspondem a A. avary ou A. brevicaudatus, duas espécies conhecidas da região, ou se formam linhagens independentes, indicando especiação críptica. As análises de WP e GMYC recuperaram cinco linhagens (espécies) no grupo interno, enquanto uma análise de bPTP delimitou três linhagens. As análises ABGD produziram dois resultados possíveis: um corroborando os métodos WP e GMYC e outro corroborando o método bPTP. Todos os métodos indicam espécies crípticas não descritas na região e apresentam variação de pelo menos uma a quatro espécies no grupo interno, dependendo da abordagem, corroborando estudos anteriores, e revelando esta região como um possível "hotspot" para descoberta de espécies de peixes não descritas.(AU)

Phylogeny and latitudinal biogeography of the Papillomaviridae family

Introduction: The latitudinal diversity gradient (LDG), a fundamental ecology pattern for higher organisms, has explained the increase of species diversity from the poles towards the tropics. Can we explain the biodiversity of Papillomaviruses based on this pattern? Objective: To analyse the phylogenetic diversity and phylogeography of most known genotypes and species belonging to Papillomaviridae Family isolated and molecularly characterized by distant latitudinal locations around the world. Methods: We collected 238 gene sequences encoding for the L1 and L2 viral capsid proteins from PaVE database. A geographical heat map based on the PV locations allowed to analyse the distribution and the number of PV species per country. Subsequently, a phylogenetic tree based on concatenated amino acid sequences L1 and L2 was constructed using a combination of Bayesian and Maximum Likelihood (ML) methods with Geneious version 11.2.1 and BEAST version 1.8.4. Statistical analysis with Principal Coordinate Analyses (PCA) using the Jaccard similarity index for presence-absence were carried out to test the similarity of the groups in the phylogenetic tree based on their viral species, the host and origin country lists. Distance-based redundancy analysis generated an ordination plot of the similarity of viral species list per group of the phylogenetic tree vs. host species per group, the country list per group and the number of samples per group. Finally, the significance of each model was tested by Analysis of variance (ANOVA). Results: A clear tendency of most of the papillomavirus (PV) species clustering in the Northern regions appeared. Our phylogenetic analyses largely support the taxonomic division into major papillomavirus genera (Alpha, Beta, Gamma, Delta, Lambda, Xi, Upsilon and Chi papillomaviruses), as reported in previous studies. Chelonid PV's, a papillomavirus in turtles, appeared as the oldest group for all papillomaviruses. Minor inconsistencies were found within some of the major taxonomic groups explained by the exhausted phylogenetic analyses. Due to the different sampling data efforts with little replication in certain groups, hosts and countries, numerical models did not fit to the proposed hypothesis. Conclusions: Phylogeographical analysis allowed determining the global distribution of papillomaviruses species diversity per latitudinal location. As most of the papillomavirus (PV) species were clustered in the Northern regions, this may clearly represent a statistical bias because larger efforts in PV studies have historically been done in the US and Europe. The biogeographical trend of a latitudinal diversity gradient does not apply for papillomaviruses. Even though no support to the phylogeographical pattern of LDG for PV was obtained, our results may only be the product of a socio-economical-scientific artefact and not a natural phenomenon.

Introducción: El gradiente de diversidad latitudinal, un patrón ecológico fundamental para los organismos superiores ha servido para explicar el incremento de la diversidad de especies desde los polos hacia los trópicos. ¿Podemos entonces explicar la biodiversidad de papilomavirus (PV) basado en este patrón? Objetivo: Analizar la diversidad filogenética y la filogeografía de los genotipos y especies más conocidas que pertenecen a la Familia Papillomaviridae aislados y molecularmente caracterizados por sus locaciones latitudinales distantes alrededor del mundo. Métodos: Recolectamos 238 secuencias de genes que codifican las proteínas de la cápside viral L1 y L2 a partir de la base de datos PaVE. Un mapa de calor geográfico basado en la ubicación de PV permitió analizar la distribución y el número de especies de PV por país. Posteriormente, un árbol filogenético basado en secuencias de aminoácidos concatenadas L1 y L2 se construyó utilizando una combinación de métodos Bayesianos y de Máxima Verosimilitud con Geneious versión 11.2.1 y BEAST versión 1.8.4. Con respecto al análisis estadístico, se realizaron tres análisis de componentes principales utilizando el índice de similitud de Jaccard para presencia-ausencia, a fin de evaluar la similitud de los grupos en el árbol filogenético en función de sus especies virales, el huésped o las listas de países. El análisis de redundancia basado en la distancia generó una gráfica de ordenación de la similitud de la lista de especies virales por grupo del árbol filogenético vs las especies hospedadoras por grupo, la lista de países por grupo y el número de muestras por grupo. Finalmente, la significancia de cada modelo se evaluó mediante análisis de varianza (ANOVA). Resultados: Existió una tendencia clara de la mayoría de las especies de papilomavirus agrupadas en regiones del norte. Nuestro análisis sustenta en gran medida la división taxonómica en los principales géneros de virus del papiloma (Alfa, Beta, Gama, Delta, Lamda, Xi, Ípsilon y Ji), como se informó en estudios anteriores. Los PV de los quelonios, papilomavirus en tortugas, apareció como el grupo más antiguo para todos los papilomavirus. Inconsistencias menores fueron encontrados dentro de algunos de los principales grupos taxonómicos explicadas por los análisis filogenéticos. Además, una serie de análisis estadísticos fueron realizados para determinar la significancia en la diversidad de especies correlacionada con la locación geográfica. Debido a diferentes esfuerzos en el muestreo de datos con poca replicación en ciertos grupos, hospedadores, y países, los modelos no se ajustaron a la hipótesis propuesta. Conclusiones: Los análisis filogeográficos permitieron determinar la distribución global de la diversidad de especies de PV por locación latitudinal. Como en la mayoría de las especies de PV fueron agrupados en las regiones del norte, esto claramente podría representar un sesgo estadístico porque esfuerzos más grandes en los estudios de PV históricamente han sido realizados en Estados Unidos de América y Europa. La tendencia biogeográfica del gradiente latitudinal de especies no aplica para los virus del papiloma. A pesar de que no se obtuvo apoyo para el patrón filogeográfico de LDG para PV, nuestros resultados podrían ser solo el producto de un artefacto social, económico y científico y no un fenómeno natural.

A new giant velvet worm from Costa Rica suggests absence of the genus Peripatus (Onychophora: Peripatidae) in Central America

Introduction: Neotropical onychophoran taxonomy and diversity has been poorly investigated. Recent studies have discovered problems in species classification: they have questioned the accepted genera and the actual number of species. This is true in Costa Rica, where several unidentified species have been reported. Objective: The objective of this investigation was to evaluate the occurrence of the accepted genera in this country, and to describe a new genus and species from Central America. Methods: In 2017, we collected one onychophoran in the Keköldi Indigenous Reserve in Talamanca, Limón, Costa Rica. The specimen gave birth to several offspring. Therefore, seven organisms were analyzed. Light microscopy was used to observe the gross morphology in all samples. The detailed morphology was studied in the biggest specimen with scanning electron microscopy; after that, we performed a phylogenetic analysis with the corresponding sequence of COI. Results: According to our results, a new genus and species of giant onychophoran was found. The genus was identified by its giant size, apical piece of seven scale ranks, large conical primary papillae, dorso-median furrow flanked by two-three accessory papillae, the absence of hyaline organs and a marked sexual dimorphism with respect to the number of legs. The new species presents a particular head pattern, as well as novel structures like cephalic papillae, accessory papillae with rudimentary apical pieces, and a lack of antennal chemoreceptors. Phylogenetic analysis rendered our genus as monophyletic and includes Peripatus solorzanoi, which is grouped within the Central American clade.As our species is clustered inside the Costa Rica-Panamanian group, it is not related to the Caribbean Island nor Guyanan Shield samples, home of Epiperipatus and Peripatus respectively. Therefore, we suggest that those genera do not occur in Central America, and a new genus exists: Mongeperipatus, gen. nov. Conclusion: We concluded that Costa Rica is home to a diversity of undescribed onychophorans that requires specific studies to help clarify the taxonomy and evolutionary relationships of the group to justify their protection.

Introducción: Los onicóforos del neotrópico han sido poco estudiados, especialmente en cuanto a taxonomía y diversidad. Sin embargo, estudios recientes sugieren que los géneros actuales son obsoletos y el número de especies existentes es desconocido. Por ejemplo, en Costa Rica, se han reportado diversos onicóforos que no han sido identificados exitosamente. Objetivo: El objetivo de este trabajo fue evaluar la presencia de los géneros aceptados en el país, y describir un nuevo género y especie para Centroamérica. Métodos: En el 2017, recolectamos un espécimen de onicóforo en la reserva indígena de Keköldi en Talamanca, Limón, Costa Rica, el cual dio a luz a varias crías. Incluyendo a este animal, estudiamos un total de siete especímenes. Mediante el uso de microscopía de luz, observamos las características morfológicas macroscópicas en todas las muestras. Asimismo, utilizamos la microscopía electrónica para analizar detalladamente la morfología en una de estas. Finalmente, realizamos un análisis filogenético con la secuencia correspondiente de COI. Resultados: Un nuevo onicóforo gigante fue descrito. Se propuso un nuevo género, el cual se distingue por su tamaño gigante, el número de filas de escamas en la pieza apical de las papilas primarias, sus grandes papilas primarias cónicas, la línea media flanqueada por dos-tres papilas accesorias, la ausencia de órganos hialinos y el dimorfismo sexual marcado respecto al número de patas. Esta especie presenta un patrón particular de papilas en su cabeza, además de estructuras nuevas como papilas modificadas en la cabeza (papilas cefálicas), papilas accesorias con piezas apicales rudimentarias y la ausencia de quimiorreceptores en las antenas. Los análisis filogenéticos lo sitúan como un género monofilético que incluye a Peripatus solorzanoi, tal clado se encuentra dentro del grupo de especies centroamericanas. Por lo tanto, la nueva especie se agrupa dentro de las muestras pertenecientes a Costa Rica-Panamá. Este grupo no se relaciona con los especímenes de las islas caribeñas ni del Escudo Guyanés, hogares de las especies tipo de Epiperipatus y Peripatus respectivamente. De modo que sugerimos que estos géneros no están presentes en Centroamérica. Así entonces, describimos el nuevo género Mongeperipatus para Costa Rica. Conclusión: Este país alberga una diversidad de onicóforos sin describir, que se requieren estudios puntuales que ayuden a aclarar la taxonomía y relaciones evolutivas del grupo para justificar la protección del filo.

Molecular Speciation and Antifungal Susceptibility Profile of Candida Species in a Tertiary Care Centre in Central Kerala.

Candidiasis is a common life-threatening condition with various clinical manifestations. It can cause significant morbidity and mortality, especially in critically ill patients. Though mainly caused by C. albicans, it has shown a change in the epidemiological pattern with an increase in the prevalence of non albicans Candida (NAC) in the recent years. This change has also reflected in the profile of antifungal susceptibility since many of these species show high level of antifungal resistance associated with treatment failures. Thus, it is important to know the regional distribution of Candida species and also find their antifungal susceptibility profile to the commonly used antifungal agents.METHODSThis is a descriptive study conducted for a period of two years on the clinical isolates of Candida species which satisfied the inclusion and exclusion criteria. Speciation was done by phenotypic methods which include germ tube test, chromogenic medium, corn meal agar and VITEK-2 system and by molecular methods using multiplex PCR. Antifungal susceptibility testing was done using automated method by VITEK-2 compact system.RESULTSAmong the 80 isolates studied, on speciation C .albicans 25(31.25%) was the most common, followed by C. tropicalis 23 (28.75%), C. parapsilosis 13 (16.25%), C. krusei 12 (15%), C. pelliculosa 4 (5%), C. auris 2 (2.5%) and C. glabrata 1 (1.25%). Most isolates were susceptible to almost all the antifungal agents tested, but C. krusei showed high level of resistance.CONCLUSIONSC. albicans was the commonest individual species, but there was an overall predominance of non-albicans Candida (NAC). Speciation of Candida isolates is important as there is wide variation in their antifungal resistance pattern. Knowledge about the prevalent species and their antifungal susceptibility will help in early initiation of appropriate treatment, thus reducing the morbidity and mortality associated with Candida infections.

Differentiation and Characterization of Mycobacterium tuberculosis Complex Causing Pulmonary Tuberculosis across North Central Nigeria

Aims: To differentiate and CharacterizeMycobacterium tuberculosiscomplex causing pulmonary tuberculosis across North Central NigeriaStudy Design:This was a simple descriptive health-based study that involved clinically suspected tuberculosis patients who were referred to two selected General Hospitals for diagnosis in each of the states across North Central Nigeria.Place and Duration of Study:This study was carried out in the North Central zone of Nigeria and it included all the seven States across North Central Nigeria using two General Hospitals Methodology:A total of 371 GeneXpert positive sputa for TB were decontaminated using N Acetyl Cysteine and Sodium Hydroxide under a level 3 Bio-safety cabinet and the resulting sediment was cultured on Lowestein-Jensen (LJ) media containing glycerol and pyruvate at 37ºC in a slanted position, SD-Bioline (TB Ag MPT 64) for the differentiation of MTBC from NTM was carried out using the isolatesfrom LJ culture. Evaluation of speciation was done using Line Probe Assay to determine the predominant species of MTBC. All the protocols used in this study followed the manufacturer’s manual strictly.Results:A total of 371 decontaminated positive GeneXpert sputa derived from 2800 suspected PTB participants were cultured on Lowestein-Jensen (LJ) medium and 302(81.40%) was found positive while 69(18.60%) were found negative. Out of the culture positive isolates, 288 (95.36%) isolates were detected on SD-BIOLINE TB Ag MPT 64 ® assay for MTBC and 14 (4.64%) as NTM. Of the 288 MTBC, three different species were identified; 272 (94.64%) were M. tuberculosis/M.Canetti,7 (2.43%) were M. africanumand 9(3.13) showed a no MTBC reaction band on all the samples that were analysed.Conclusion: Differentiations of MTBC from NTM has help to re-confirm that not all symptoms of pulmonary infection are caused by MTBC but NTM are implicated due to their distribution in the environment, however, molecular characterisation method has narrow our findingsdown to M.tuberculosis/M.canettiias the predominant specie of MTBC circulating in the region, although, M.africaumwas also detected and these two species of MTBC are the leading cause of pulmonary tuberculosis across all the North Central state of Nigeria.per state. The study included 371 positive sputum samples drawn from 2800 suspected pulmonary TB patients between 2017 and 2018.Original ResearchArticle

Determination of arsenic speciation in 17 commonly used traditional Chinese herbal medicines by HPLC-ICP-MS / 中国中药杂志

The element speciation analysis for heavy metals in herbal medicines is still in the beginning stage. In this study,the total amount of arsenic( As) in 103 batches of 17 commonly used Chinese medicines( including 16 plant medicines and 1 medicinal fungus) was detected by inductively coupled plasma mass spectrometry( ICP-MS). Furthermore,based on HPLC-ICP-MS,the simultaneous detection methods of six As speciation kinds in traditional Chinese medicines were established. An AS7 anion exchange column was selected and the As forms in 17 traditional Chinese medicines was systematically analyzed. The results showed that the method of pretreatment of medicinal materials by microwave digestion and the detection of total amount of As by ICP-MS was stable and reliable. As for the speciation analysis of As,the high-speed ultrasonic extraction method was adopted,and it showed that the linear relationship of the six As speciation was satisfied with the correlation coefficient R2>0. 999 9. The LOQ of six kinds of As speciation were 0. 20,0. 10,0. 15,0. 10,0. 25,0. 10 μg·L~(-1) for arsenic betaine( As B),arsenious acid [As( Ⅲ) ],dimethyl arsenic( DMA),arsenic choline( As C),monomethyl arsenic( MMA),arsenic acid[As( Ⅴ) ],respectively. The recoveries were between 84. 24% and 121. 5%,and the relative standard deviations were 2. 7% to 11%. Among the 103 batches of medicinal materials,only one batch of sample As exceeded the Chinese Pharmacopoeia limit standard; As( Ⅲ) and As( Ⅴ) had high detection rate in 103 batches of Chinese herbal medicines,within which As( Ⅴ) was the main detected form,and inorganic As accounted for the ratio reached 80. 90%-98. 73%; some samples detected DMA,MMA and As B,As C was not detected in any batch. This study established an analytical method suitable for the speciation of As in Chinese herbal medicines,and provided basic data for As residual residue in Chinese herbal medicines,which can provide important reference for the risk assessment and quality standards.

Speciation analysis of trace mercury and arsenic in 31 kinds of animal drugs and discussion about the limit standards / 药学学报

The toxicity of heavy metals and harmful elements is close related to their speciation. In the present study, the methods for mercury and arsenic speciation analysis based on high-performance liquid chromatography conjunction with inductively coupled plasma mass spectrometry (HPLC-ICP-MS) were established and applied to the determination of 31 kinds of animal drugs, 29 of which were included in the Chinese Pharmacopeia (2015 edition). The results showed that the LODs for all the speciation were within 0.1-0.65 μg·kg-1, and the recoveries were within 86.9%-116.6% with the RSD of 1.49%-4.23%. Inorganic mercury (Hg2+) was detected in all the 87 batches of samples that came from 31 kinds of animal drugs, and the contents were 2.39-6567 μg·kg-1. Methylmercury (MeHg) was detected in 33 batches of samples that came from 12 kinds of animal drugs, and the contents were 2.83-319.7 μg·kg-1. Ethylmercury (EtHg) were detected in none of the samples. The detection rates of As(Ⅲ), As(V), monomethylarsononous acid (MMA), dimethylarsinic acid (DMA), arsenobetaine (AsB) and arsenocholine (AsC) in the 31 batches of animal drugs was 96.77%, 100%, 45.16%, 90.32%, 93.55% and 22.58%, respectively. According to the toxic level of different speciation, the animal drugs with high risks of mercury were Agkistrodon, Bungarus Parvus, Zaocys, and Scolopendra; the animal drugs with high risks were Pheretima, Agkistrodon, Zaocys, and Aspongopus. This study can provide important evidence for the risk assessment, setting and revision of the limit standards of heavy metals and harmful elements.

Research progress of chromium and its speciation analysis / 中国中药杂志

Chromium has been discovered for more than two hundred years, and it has advantages of wear-resisting, high temperature, corrosion resistance and so on. Chromium has been widely used in industrial production and received extensive attention in China and abroad. Detailed limit standards have been set for chromium in food and fishery product, and chromium can also be enriched in many traditional Chinese medicines. Besides, the toxicities of different chromium speciation are quite different, and thus morphological analysis is necessary. However, the transformation or migration is easily happened among different speciation which brings difficulties to research and analysis. This article summarizes the research status of chromium and its aims to provide reference for scientific research and pollution prevention of chromium.

Estimation of genetic variation in closely related cycad species in Ceratozamia (Zamiaceae: Cycadales) using rapds markers / Variación genética en especies de cícadas estrechamente relacionadas en Ceratozamia (Zamiaceae: Cycadales) mediante el uso de RAPDs

AbstractThe Ceratozamianorstogii complex from Southern Mexico is made up of four closely related taxa and occurs in similar habitats (Quercus forest). All have linear-lanceolate leaflets with great similarity between them, especially in juvenile stages, but differentiate with age. There has been debate regarding delimitation of species due to character loss in herbarium specimens. The aim of this study was to determine the genetic variation, and to measure genetic similarity between the four taxa. We studied populations in Cintalapa (Chiapas) for C. alvarezii and C. norstogii; the Sierra Atravesada (Oaxaca) for C. chimalapensis, and Villa Flores (Chiapas) for C. mirandae. One population for each taxon was sampled (only one population is known for C. alvarezii) 11-15 randomly chosen adult individuals were sampled. Twenty-eight primers were tested of which five were polymorphic using the RAPD'S technique. The data were analyzed using Bayesian methods. Results revealed low genetic diversity, and a differentiation was found between species, suggesting a recent divergence. A previous morphological and anatomical study on the complex has found the taxa to be distinct. However, the results of this study have shown that the C. norstogii species complex is in a divergence process, probably through genetic drift and founder effects. Rev. Biol. Trop. 65 (1): 305-319. Epub 2017 March 01.

ResumenLos cuatro taxa que componen el complejo Ceratozamia norstogii de especies en el sur de México están estrechamente relacionados y se dan en hábitats similares (bosque de Quercus). Todos tienen folíolos linear-lanceolados con gran similitud entre ellos, sobre todo en las etapas juveniles, pero se diferencian con la edad. Ha habido un debate en relación con la delimitación de especies debido a la pérdida de caracteres en especímenes de herbario. Los objetivos de este estudio son determinar la variación genética y medir la similitud genética entre los cuatro taxones en el complejo. Las poblaciones estudiadas están en; Cintalapa, Chiapas para C. alvarezii y C. norstogii, la Sierra Atravesada, Oaxaca para C. chimalapensis y Villa Flores, Chiapas para C. mirandae. Se tomaron muestras de una población de cada taxón (sólo una población es conocida para C. alvarezii) 11-15 individuos adultos elegidos al azar fueron muestreados. Veintiocho primers fueron probados, de los cuales cinco fueron polimórficos mediante la técnica RAPD's. Los datos fueron analizados utilizando métodos bayesianos. Los resultados revelaron baja diversidad genética y la diferenciación encontrada entre las especies sugiere una divergencia reciente. Un estudio morfológico y anatómico anterior en el complejo encontró que los taxa son distintos. Sin embargo, los resultados del presente estudio han demostrado que el complejo C. norstogii aun se encuentra en un proceso de divergencia, probablemente a través de deriva genética y efectos de fundador.

The current status of the Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae) species complex

Lutzomyia longipalpis s.l. is a complex of sibling species and is the principal vector of American visceral leishmaniasis. The present review summarises the diversity of efforts that have been undertaken to elucidate the number of unnamed species in this species complex and the phylogenetic relationships among them. A wide variety of evidence, including chemical, behavioral and molecular traits, suggests very recent speciation events and complex population structure in this group. Although significant advances have been achieved to date, differential vector capacity and the correlation between structure of parasite and vector populations have yet to be elucidated. Furthermore, increased knowledge about recent epidemiological changes, such as urbanisation, is essential for pursuing effective strategies for sandfly control in the New World.

DNA Barcoding unveils cryptic lineages of Hoplias malabaricus from Northeastern Brazil

Abstract The trahira or wolf fish - Hoplias malabaricus- is a valid species, although recent cytogenetic and molecular studies have indicated the existence of a species complex. In this context, the present study analyzed the mitochondrial COI marker to determine the levels of genetic diversity of specimens from the Brazilian state of Maranhão, and verify the occurrence of distinct lineages within the study area. Samples were collected from the basins of the Turiaçu, Pindaré, Mearim, Itapecuru, and Parnaíba rivers. A 630-bp fragment was obtained from 211 specimens, with 484 conserved and 108 variable sites, and 60 haplotypes (Hd = 0,947; = 0,033). The phylogenetic analyses indicated the existence of three distinct lineages of H. malabaricus from Maranhão. Genetic distances of 1.5-8.2% were found between all the populations analyzed, while the variation between haplogroups ranged from 2.1% to 7.7%. The AMOVA indicated that most of the molecular variation was found among groups, with high FST values. The high levels of genetic variability found in the present study are supported by the available cytogenetic data. These findings reinforce the need for the development of effective programs of conservation and management independently for each river basin, in order to preserve the genetic variability found in this taxon.

Resumo A traíra - Hoplias malabaricus- é uma espécie válida, embora recentes estudos citogenéticos e moleculares tenham indicado a existência de um complexo de espécies. Neste contexto, o presente estudo analisou o marcador mitocondrial COI para determinar os níveis de diversidade genética dos espécimes do estado do Maranhão e verificar a ocorrência de linhagens distintas dentro da área de estudo. As amostras foram coletadas nas bacias dos rios Turiaçu, Pindaré, Mearim, Itapecuru e Parnaíba. As análises filogenéticas indicaram a existência de três linhagens distintas nas populações do Maranhão. Obteve-se um fragmento de 630 pb de 211 espécimes, com 484 sítios conservados, 108 variáveis e 60 haplótipos (Hd = 0,947; = 0,033). As análises filogenéticas indicaram a ocorrência de três linhagens distintas de H. malabaricus do Maranhão. Distâncias genéticas de 1.5 a 8.2% foram encontradas entre todas as populações analisadas, enquanto a variação entre os haplogrupos variou de 2.1% a 7.7%. A AMOVA indicou que a maior variação molecular foi entre os grupos, com altos valores de FST. Os altos níveis de variabilidade genética encontrados no presente estudo são suportados pelos dados citogenéticos disponíveis. Essas descobertas reforçam a necessidade de desenvolver programas de conservação e manejo independentemente para cada bacia hidrográfica, a fim de preservar a variabilidade genética encontrada neste táxon.

Study on Copper Absorption, Accumulation and Tolerance Mechanism of Alfalfa / 分析化学

To reveal the mechanism of Cu enrichment of alfalfa, in-situ micro-X-ray fluorescence spectrometry (μ-XRF) and fractional extraction were used to explore in-situ Cu distribution information in alfalfa seedlings and different combination forms of Cu in organs.The results showed that alfalfa roots were enriched with Cu up to 12.06 mg/g, which was 8 times of stem and 4.9 times of leaves.The in-situ μ-XRF result showed that the root of alfalfa was the main site of enrichment of Cu, and there was a barrier of Cu at the rhizome junction to alleviate the toxic effect of excess Cu on the shoots.Excess Cu (more than 50 μmol/L) also inhibited the uptake of Zn and Ca in alfalfa, enhanced the uptake of Fe in alfalfa, but had no obvious effect on the uptake of K and Mn.Plant fractional extraction showed that the Cu in root cells was mainly fixed in the form of insoluble residues (41%) and cell wall chelate (20%), while in the stem, Cu existed as four forms including hydrophobic protein binding, cell wall binding state, residual state and water-soluble state, which further reduce the excess transport of Cu to the leaves.In the leaves, excess Cu in the leaf cells was mainly in the vacuole and insoluble residue to achieve tolerance and detoxification to Cu.

Distribution and Speciation of Lead in Moss Collected from a Lead-Zinc Mining Area by Micro-X-Ray Fluorescence and X-Ray Absorption Near Edge Structure Analysis / 分析化学

Moss has high tolerance and accumulating capacity to heavy metal.In this study, the distribution of heavy metal elements in moss sampled from lead-zinc mine was analyzed by X-ray fluorescence spectrometry.The speciation of lead was analyzed by X-ray absorption near-edge spectroscopy.Research showed that the contents of Pb, Zn, Cd and As in the moss of the mining area were extremely high, and their maximum concentration were 1.06 mg/g , 1.23 mg/g, 30.5 μg/g, 13.2 μg/g, respectively.Besides, the shoots especially the new tissue of the moss were the major sites for accumulation and storage of heavy metals.The micro-distribution characteristics varied among Hypnum plumaeforme and Brachytheciumprocumbens, indicating the difference of different species of moss in absorption pathway, accumulation and tolerance mechanisms for heavy metal.Linear combination fitting results indicated that the main lead speciation in moss was lead phosphate (78%) and lead oxide (22%), which suggested that the precipitation of lead phosphate might be the main mechanism of tolerance for moss.

Comparison of Speciation Analysis of 7 Trace Elements in Opercalina turperthum by Water Extraction and Semi-bionic Extraction / 中国药房

OBJECTIVE:To study the extraction rate and speciation of antivitiligo-related elements in Opercalina turperthum by water extraction and semi-bionic extraction. METHODS:Water extraction and semi-bionic extraction were respectively used. Wa-ter-soluble state and suspension state in extract of O. turperthum were separated by microporous filtering film;organic and inorgan-ic trace elements in water-soluble state were separated by macroporous resin. The contents of Cu,Zn,Fe,Ca,Mg,Mn and Sr were detected by flame atomic adsorption spectrophotometry,and speciation analysis was conducted. RESULTS:After water extrac-tion,extraction rates of 7 trace elements were 40.47%-72.49%;ratio of suspended particles was 3.69%-8.78%;ratio of organic state/inorganic state was 104.36% in water-soluble state of Sr and 3.94%-48.39% in water-soluble state of Cu,Zn,Fe,Ca,Mg, Mn. After semi-bionic extraction,except for Mn,extraction rates of trace elements were higher than water extraction,extraction rates of Cu,Zn,Fe,Ca,Mg,Sr were 77.69%-90.19%;ratio of suspended particles was 0.39%-8.57%;the ratio of organic state/inorganic state was 72.74%-180.79% in water-soluble state of elements. CONCLUSIONS:Cu,Zn,Fe,Ca,Mg,Mn and Sr in O. turperthum are dissolved easily,mainly existing in the form of inorganic state. After semi-bionic extraction,except for Mn,the dis-solution rate of other elements and the proportion of organic trace elements after dissolution increase significantly. Both dissolution and effective utilization rate of trace elements by semi-bionic extraction are higher than water extraction.

Influence of different processing methods on contents and speciation of heavy metals in Cortex Eucommiae / 国际药学研究杂志

Objective To investigate the influence of three different processing methods on contents and speciation of heavy metals Pb, Cd, Hg and Cu and a harmful nonmetallic element As in Cortex Eucommiae. Methods The five elements were measured by atomic absorption spectrometry. The determination wavelengthes of Pb and Cd was 283.3 and 228.8 nm using graphite furnace atomic absorption spectrometry; the determination wavelength of Cu was 324.8 nm using flame atomic absorption spectrometry; the determination wavelengthes of As and Hg were 193.7 and 253.6 nm using flow injection-hydride generation method. Results Heavy metal’s contents in Cortex Eucommiae stir-fried with salt were increased. Contents of heavy metals in Cortex Eucommiae stir-fried and baked were decreased. Heavy metal morphology had no differences between processed products and raw materials. The experimental results showed that main existing form of Pb was in organic matter-bound, and mercury existed in exchange condition and organic matterbound. Copper existed in organic matter-bound and carbonate bound and arsenic mainly existed with residual forms. Cadmium mainly exited in carbonate exchange condition. Conclusion There is a certain effect on the heavy metal content but less effect on the form of heavy metals by different processing methods.

Phylogenetic analysis reveals cryptic species diversity within minute intestinal fluke, Stellantchasmus falcatus Onji and Nishio, 1916 (Trematoda, Heterophyidae)

Objective To examine the taxonomic boundaries in the Stellantchasmus populations in different hosts and reveal the cryptic speciation of the Stellantchasmus in Thailand based on both morphological and molecular approaches. Methods Stellantchasmus falcatus (S. falcatus) s.l. was collected from different hosts throughout Thailand. The worms were examined and measured. The nuclear ITS2 gene and the mitochondrial COI gene were used to investigate the S. falcatus s.l. from Dermogenus pusillus and Liza subviridis. Results Stellantchasmus was one of food-borne trematode that widely distribute throughout Thailand. The comparison of S. falcatus s.l. from Dermogenus pusillus and Liza subviridis indicated a genetic divergence of nuDNA with 3.6% and mtDNA with 19.3%, respectively. Morphological characteristics were fairly different in the ratio of body size, length of prepharynx, and ratio of organ size. Conclusions Our results provide initial evidence that S. falcatus s.l. from different hosts tend to be a different species based on both molecular and morphological characters. Cryptic species complexes are generally found among parasites that tend to have large populations and/or rapid evolution. The degree of genetic diversity existing would suggest the practice of targeted regimes to design and minimize these lections of anthelmintic resistance in the future.