A molecular and morphological study of Ascaris suum in a human-pig contact scenario in northeastern Brazil / Estudo molecular e morfológico de Ascaris suum em um cenário de contato entre suínos e humanos no nordeste brasileiro

The aim of the present study was to assess morphologic and genetic data on ascariasis in swine (Sus scrofa domesticus) and humans in low-resource rural and periurban communities in the state of Piauí, Brazil. Our cross-sectional survey included 100 fecal samples obtained from swine and 682 samples from humans. Fifteen pigs were necropsied. Human and porcine fecal samples were examined to identify Ascaris eggs. Parasites obtained in the swine necropsies were studied using scanning electron microscopy (SEM), and the mitochondrial gene encoding the cytochrome oxidase 1 (cox1) enzyme was partially amplified and sequenced for molecular taxonomy and phylogenetic analyses. The overall prevalence of Ascaris eggs in the swine fecal samples was 16/100 (16%). No Ascaris eggs were identified in the human fecal samples. SEM of six worms recovered from pigs demonstrated morphological characteristics of A. suum. Cox1 sequences were compatible with A. suum reference sequences. Original and reference (GenBank) nucleotide sequences were organized into clusters that did not segregate the parasites by host species or and region. The largest haplogroups were dominated by haplotypes H01, H02 and H31. In the communities studied, there was no epidemiological evidence of the zoonotic transmission of ascariasis at the human-swine interface.(AU)

O presente estudo teve como objetivo acessar dados morfológicos e genéticos sobre a ascaridíase em suínos (Sus scrofa domesticus) e humanos, em comunidades rurais e periurbanas no estado do Piauí. O estudo transversal incluiu 100 amostras fecais de suínos e 682 amostras obtidas de humanos. Quinze suínos foram necropsiados. Amostras fecais suínas e humanas foram examinadas para detecção de ovos de Ascaris. Os parasitas adultos, obtidos nas necropsias, foram estudados através de microscopia eletrônica de varredura (MEV), e o gene mitocondrial codificante da enzima citocromo oxidase 1 (cox1) foi parcialmente amplificado e sequenciado para análises filogenéticas e de taxonomia molecular. A prevalência de Ascaris em amostras fecais de suínos foi 16/100 (16%), não sendo identificado nenhum caso de infecção por este parasita em humanos. A análise por MEV de parasitas recuperados de suínos demonstrou características morfológicas de Ascaris suum. As sequências nucleotídicas de cox1 foram compatíveis com A. suum. As sequências originais e de referência (obtidas no GeneBank) foram organizadas em clusters que não segregaram os parasitas por hospedeiro ou região geográfica. Os maiores haplogrupos foram dominados pelos haplótipos H01, H02 e H31. Nas comunidades estudadas, não foi evidenciada transmissão zoonótica de A. suum na interface suíno-humana.(AU)

Taxonomia integrativa de espécies da série Shannoni de Psathyromyia (Psathyromyia) Barretto, 1962 (Diptera, Psychodidae, Phlebotominae) do estado de São Paulo / Integrative taxonomy of Psathyromyia (Psathyromyia) Barretto, 1962 Shannoni series species (Diptera, Psychodidae, Phlebotominae) from São Paulo state

Os flebotomíneos são dípteros que habitam ambientes florestais e antrópicos e são encontrados comumente em cavernas. Com exceção da Antártica, habitam todos os demais continentes. Atualmente o grupo é composto por pouco mais de 1.000 espécies. As fêmeas são hematófagas e várias espécies apresentam relevância em saúde pública como vetores de agentes de leishmanioses, arboviroses e bartonelose. A taxonomia dos flebotomíneos vem se desenvolvendo desde o início do século XX amplamente baseada em estudos morfológicos e morfométricos lineares das formas adultas. Atualmente tem sido recomendada a abordagem integrativa na delimitação das espécies, que além dos estudos clássicos, utiliza informações moleculares, das formas imaturas, geometria e ecologia, além de outros parâmetros. O objetivo deste projeto foi contribuir para um melhor conhecimento das quatro espécies do subgênero Psathyromyia que ocorrem no estado de São Paulo, duas das quais foram recentemente descritas (Pa. baratai e Pa. ribeirensis) e as duas outras revalidadas (Pa. bigeniculata e Pa. limai), a fim de encontrar diferenças entre as fêmeas dessas espécies, morfologicamente muito semelhantes. Coleções entomológicas foram consultadas, e para a coleta de espécimes foram amostrados ambientes florestais e antrópicos do estado de São Paulo e Mato Grosso do Sul, utilizando diferentes armadilhas de coleta. Foram realizadas análises discriminantes na morfometria linear de estruturas da cabeça e na morfometria geométrica das asas, além de análises moleculares do fragmento do gene citocromo c oxidase subunidade I (COI) por meio da construção de uma árvore de Neighbor-Joining (NJ) e método de delimitação de espécies ABGD (Automatic Barcode Gap Discovery), com a inclusão de sequências de Pa. abonnenci, Pa. bigeniculata e Pa. shannoni depositadas no GenBank. Como resultados, as análises discriminantes da morfometria linear possibilitaram a separação das espécies, exceto o par Pa. ribeirensis/Pa. limai. Para a morfometria geométrica, o tamanho do centroide evidenciou diferenças entre as asas de Pa. limai e Pa. ribeirensis em relação à de Pa. bigeniculata e a análise de variância de procrustes evidenciou a separação de todas as espécies, mesmo após a validação cruzada, exceto entre Pa. ribeirensis e Pa. limai, sugerindo que essas duas espécies são indistinguíveis morfometricamente. As análises do fragmento do gene COI foram úteis para separar as espécies. Psthyromyia bigeniculata apresentou altos valores de distância intraespecífica (4,7%), que pode representar diversidade críptica. De modo geral, a taxonomia integrativa foi útil para separar três das quatro espécies estudadas e corrobora a retirada de Pa. limai e Pa. bigeniculata da sinonímia de Pa. shannoni, bem como a descrição de Pa. baratai como um táxon diferente. Fêmeas de Pa. ribeirensis permanecem indistinguíveis daquelas de Pa. limai pela morfologia e morfometrias linear e geométrica.

Sand flies are dipterans that inhabit forest and anthropic environments and are commonly found in caves. Except for Antarctica, they inhabit all other continents. Currently the group comprises about 1,000 species. Females are hematophagous and several species are relevant for public health as leishmaniasis, arboviruses and bartonellosis agent vectors. The taxonomy of sand flies has been developing since the beginning of the 20th century, largely based on morphological and linear morphometric studies of the adult forms. Currently, an integrative approach to species delimitation has been recommended, which, in addition to classical studies, uses molecular information, immature forms, geometry and ecology, among other parameters. The aim of this project was to contribute to a better understanding of the four species of the Psathyromyia subgenus that occur in the state of São Paulo, two of them were recently described (Pa. baratai and Pa. ribeirensis) and the two others revalidated (Pa. bigeniculata and Pa. limai), in order to find differences between the females of the species, which have similar morphological traits. Museum entomological collections were consulted, and wild-caught specimens were collected in forest and anthropic environments in the states of São Paulo and Mato Grosso do Sul, using different collection traps. Discriminant analyzes were performed on the linear morphometry of head structures and on geometric morphometry of wing, in addition to molecular analyzes of the cytochrome c oxidase subunit I (COI) gene fragment by means of the construction of a Neighbor-Joining (NJ) tree and species delimitation method ABGD (Automatic Barcode Gap Discovery), wth the inclusion of Pa. abonnenci, Pa. bigeniculata and Pa. shannoni sequences deposited in GenBank. As a result, the discriminant analyzes of linear morphometry allowed the separation of species, except for the pair Pa. ribeirensis/Pa. limai. For the geometric morphometry, the centroid size showed differences between wings of Pa. limai and Pa. ribeirensis in relation to that of Pa. bigeniculata, while the procrustes analysis of variance showed the separation of all species even after cross-validation, except between Pa. ribeirensis and Pa. limai, suggesting that these two species are morphometrically indistinguishable. COI gene fragment analyzes were useful to separate the species. Psthyromyia bigeniculata showed high intraspecific distance values (4.7%), which may represent cryptic diversity. In general, the integrative taxonomy was useful to separate three of the four species studied and corroborates the resurrection of Pa. limai and Pa. bigeniculata from the synonymy of Pa. shannoni, as well as the description of Pa. baratai as a different taxon. Females of Pa. ribeirensis remain indistinguishable from those of Pa. limai for linear and geometric morphology and morphology.

DNA barcoding revels first records of three rare coleopteran genera in Northern lakes of Egypt / O código de barras de DNA revela os primeiros registros de três gêneros raros de coleópteros nos lagos do norte do Egito

Abstract One aquatic coleopteran species from family Dytiscidae and two aquatic coleopteran genera from family Hydrophilidae were recorded in the summer period and represent first records in the Egyptian lakes. Beetles were collected from two northern lakes, Lake Idku and Lake Burullus. They were identified by morphological characteristics as well as the mtDNA barcoding method. A molecular phylogenetic approach was used to determine the genetic identity of the collected samples based on the mitochondrial cytochrome oxidase I (COI). Prodaticus servillianus (Dytiscidae) from Egypt showed no significant difference in the COI region and they are highly similar to P. servillianus from Madagascar. The phylogenetic analysis revealed that the other two coleopteran genera belong to family Hydrophilidae. Based on COI only, there is no clear evidence for their genetic identity at the species level. So, we defined them to the closest taxon and denoted them as Cymbiodyta type A and B. The results indicated that resolving the molecular identity of the aquatic beetles from northern lakes of Egypt need more considerations in the field of biological conservation. We concluded that utilization of COI as a barcoding region for identifying some coleopteran species is not sufficient and additional molecular markers are required to uncover the molecular taxonomy at deep levels.

Resumo Uma espécie de coleópteros aquático da família Dytiscidae e dois gêneros de coleópteros aquáticos da família Hydrophilidae foram registrados no período de verão e representam os primeiros registros nos lagos egípcios. Os besouros foram coletados em dois lagos do norte, o lago Idku e o lago Burullus, e identificados por características morfológicas e pelo método de código de barras mtDNA. Uma abordagem filogenética molecular foi usada para determinar a identidade genética das amostras coletadas com base no citocromo oxidase I mitocondrial (COI). Prodaticus servillianus (Dytiscidae) do Egito não mostrou diferença significativa na região COI e é altamente semelhante a P. servillianus de Madagascar. A análise filogenética revelou que os outros dois gêneros de coleópteros pertencem à família Hydrophilidae. Com base apenas no COI, não há evidências claras de sua identidade genética no nível da espécie. Assim, nós os agrupamos no táxon mais próximo e os denominamos Cymbiodyta tipo A e B. Os resultados indicaram que a identidade molecular dos besouros aquáticos dos lagos do norte do Egito precisa de mais considerações no campo da conservação biológica. Concluímos que a utilização de COI como região de código de barras para identificar algumas espécies de coleópteros não é suficiente, sendo necessários marcadores moleculares adicionais para descobrir a taxonomia molecular em níveis profundos.

Avistamientos de aves y análisis bioinformático en entornos rurales con estudiantes de la Institución educativa Ferralarada en Choachí (Colombia) / Bird sightings and bioinformatic analysis in rural settings with students from the educational institution Ferralarada in Choachí (Colombia)

Objetivo: La ejecución de esta investigación presenta dos objetivos principales: primero. Realizar avistamientos de aves en entornos rurales; con estudiantes de primaria, por medio del dibujo como herramienta descriptiva. Segundo. Desarrollar una metodología sencilla para realizar un análisis bioinformático rápido, utilizando el criterio de parsimonia. Materiales y métodos: El presente trabajo se desarrolló con la comunidad educativa de la Institución Educativa Departamental Técnica Agropecuaria Ferralarada; ubicada en el municipio de Choachí, Colombia. Como respuesta a la necesidad de continuar los procesos pedagógicos de la institución en materia ambiental y a los retos actuales de la educación presentados por la emergencia sanitaria del COVID-19. Se realizó una serie de actividades experimentales, que surgen de las aves como elemento motivacional. Resultados. Las actividades realizadas por los docentes de primaria a propósito de los avistamientos, han generado registros de dibujo y escritos, los cuales expresan las representaciones que tienen los estudiantes a propósito de su entorno natural. Se construyo un árbol filogenético basado en el criterio de parsimonia utilizando secuencias del gen CO1 extraídas de Genbank, y analizadas con los programas ClustalX, Nona y Asado. Conclusión: Las actividades ejecutadas a propósito de los avistamientos, han resultado ser viables y de fácil desarrollo con la comunidad educativa. La propuesta bioinformática se configura como una herramienta de análisis sencillo con validez procedimental, sin embargo, requiere avances conceptuales frente al manejo de programas y plataformas, así como disponibilidad de recursos tecnológicos para su implementación.

Aim: The execution of this proposal has two main aim: 1. Perform bird watching in rural settings; with primary school students, using drawing as a descriptive tool. 2. Develop a simple methodology to perform a rapid bioinformatic analysis, using the parsimony criterion. Materials and methods: This work was developed with the educational community of the Institución Educativa Departamental Técnica Agropecuaria Ferralarada; located in the municipality of Choachí, Colombia. In response to the need to continue the pedagogical processes of the institution in environmental matters and to the current challenges of education presented by the health emergency of COVID-19. A series of experimental activities were carried out, arising from birds as a motivational element. Results. The activities carried out by the primary school teachers regarding the sightings have generated drawing and written records, which express the representations that students have of their natural environment. A phylogenetic tree was built based on the parsimony criterion using sequences of the CO1 gene extracted from Genbank, and analyzed with the ClustalX, Nona and Asado programs. Conclusion: The activities carried out in connection with the sightings have turned out to be viable and easy to develop with the educational community. The bioinformatics proposal is configured as a simple analysis tool with procedural validity, however, it requires conceptual advances in the management of programs and platforms, as well as the availability of technological resources for its implementation.

Genetic variability of populations of the white-eared opossum, Didelphis albiventris Lund 1840 (Didelphimorphia; Didelphidae) in Brazil / Variabilidade genética de populações de gambá de orelha branca, Didelphis albiventris Lund 1840 (Didelphimorphia; Didelphidae) no Brasil

Abstract Didelphis albiventris are found throughout Northeast and Central Brazil to central-southern Uruguay and it was subject of few studies in a population level. Given this, the present study investigated the genetic variability of the species using the mitochondrial molecular marker cytochrome oxidase c subunit I. We analyzed samples from the different biomes within three Brazilian regions: Northeast (Caatinga , Cerrado, and Atlantic Forest), Southeast (Cerrado , Atlantic Forest, Cerrado/Atlantic Forest, and Cerrado/Caatinga ecotones) and South (Pampa and Atlantic Forest). Software BAPs retrieved five distinct demes: dm 1, dm 2, and dm 5 that occurs in South, Northeast and Southeast regions respectively and the dm 3 and dm 4 are wide distributed in Northeast and Southeast. Population analysis performed with AMOVA, haplotype network and Mantel test estimated the veracity of the demes. The FST shows structuring for the five demes, with dm 1 (South region) isolated from the others, however the other analysis showed the Northeast/Southeast demes (dm 2-5) united, diagnosing gene flow between them, mainly at the transitional zones, in areas as far away as areas with similar latitude interval (Southeast vs South) that was not detected gene flow. In the haplotype network, the mutational steps was conclusive in split dm1 from dm 2-5 with 15 mutational steps and the Mantel test was moderated, which is explained by genetic similarity despite the great geographic distances (Northeast/Southeast). Thus, our analysis recognized two different lineages (South and Northeast/Southeast) and indicate that the biomes were not decisive in their isolation. The sharing of demes at the transitional zones and in areas with high latitudinal intervals reflects a recent ancestral polymorphism for D. albiventris. The plasticity in the occupation of the space by this species contributes in its wide dispersion capability, that is, geographical distribution. Our results revealed important implications for the management of D. albiventris in these transitional zones areas where demes were shared.

Resumo Didelphis albiventris é encontrada em todo o Nordeste e região central do Brasil até o centro-sul do Uruguai e foi alvo de poucos estudos em nível populacional. Dessa forma, o presente estudo, investiga a variabilidade genética da espécie usando o marcador molecular citocromo c oxidase subunidade I. Analisou-se amostras de diferentes biomas de três regiões brasileiras: Nordeste (Caatinga, Cerrado e Floresta Atlântica), Sudeste (Cerrado, Floresta Atlântica, ecótonos Cerrado/Floresta Atlântica e Cerrado/Caatinga) e Sul (Pampa e Floresta Atlântica). O software BAPs recuperou cinco demes distintos: dm 1, dm 2 e dm 5, que ocorrem nas regiões Sul, Nordeste e Sudeste, respectivamente, e os dm 3 e dm 4, que são amplamente distribuído no Nordeste e Sudeste. Análises populacionais realizadas com AMOVA, rede de haplótipo e teste de Mantel estimaram a veracidade das demes. O FST mostrou estruturação para as cinco demes, com dm 1 (região Sul) isolada das demais, entretanto as outras análises mostraram as demes Nordeste/Sudeste (dm 2-5) unidos, diagnosticando fluxo gênico entre elas, principalmente em zonas de transição, em áreas tão distante quanto áreas com similar intervalo de latitude (Sudeste e Sul), onde não foram detectado fluxo gênico. Na rede de haplótipo, os passos mutacionais foram conclusivos em separar dm 1 do dm 2-5 com 15 passos mutacionais, e o teste de Mantel foi moderado, o que é explicado pela similaridade genética apesar da grande distância geográfica (Nordeste/Sudeste). Assim, duas linhagens diferentes (Sul e Sudeste/Nordeste) foram encontradas, indicando que os biomas não foram decisivos em seus isolamentos. Os compartilhamentos das demes, em zonas de transição e em áreas com elevados intervalos de latitude, refletem um polimorfismo ancestral recente para D. albiventris. A plasticidade na ocupação do espaço por esta espécie contribui em sua ampla capacidade de dispersão, ou seja, distribuição geográfica. Nossos resultados revelam importantes implicações para o manejo de D. albiventris nessas áreas de zonas de transição, onde as demes são compartilhadas.

Molecular phylogenetic analysis of Cysticercus ovis from Egypt based on MT-CO1 gene sequences / Análise filogenética molecular de Cysticercus ovis do Egito com base nas sequências do gene MT-CO1

Abstract Cysticercus ovis or sheep measles is the larval stage of Taenia ovis, which is the intestinal tapeworm of dogs. It is found in the cardiac and skeletal muscles of sheep and can be the cause of partial or total condemnation of carcasses at abattoirs. The aim of the current work was to determine the prevalence of C. ovis among sheep in Upper Egypt and to present the molecular and phylogenetic analysis of this using the amplified Mitochondrial Cytochrome Oxidase subunit 1 (MT-CO1) gene. A total of 1885 sheep slaughtered at local abattoirs of 4 different governorates of Upper Egypt (Asuit, Sohag, Qena and Aswan) were carefully examined for C. ovis. The overall prevalence of infection was 2.02%. The highest rate of infection was observed in adult animals over 4 years of age (44.73%). There was no significant effect of animal sex on infection rates. The phylogenic analysis of C. ovis Egyptian isolates showed very close similarity to the New Zealand isolate (AB731675). This is the first report showing the genetic analysis of C. ovis in Egypt, which provides a very powerful tool for taxonomy and definitive diagnosis of C. ovis, which could be helpful for preventive and control programs.

Resumo Cysticercus ovis "sheep measles" é o estágio larval da Taenia ovis, encontrada nos músculos de carneiros, causado pela ingestão de ovos de Taenia ovis, parasita de cães. O objetivo do presente trabalho foi determinar a prevalência de C. ovis entre ovinos no Alto Egito e apresentar as análises moleculares e filogenéticas, utilizando o gene da subunidade mitocondrial citocromo-oxidase amplificada 1 (MT-CO1). Um total de 1885 ovinos abatidos em matadouros locais de 4 províncias diferentes do Alto Egito (Asuit, Sohag, Qena e Aswan) foram cuidadosamente examinados para C. ovis. A prevalência geral de infecção foi de 2,02%. A maior taxa de infecção foi observada em animais adultos com mais de 4 anos de idade (44,73%). Não houve efeito significativo do sexo nas taxas de infecção. A análise filogenética de isolados egípcios de C. ovis mostrou uma similaridade muito próxima ao isolado da Nova Zelândia (AB731675). Este é o primeiro relato mostrando a análise genética de C. ovis no Egito, fornecendo uma ferramenta para taxonomia e diagnóstico definitivo de C. ovis, podendo ser útil para programas preventivo e de controle.

Identity of Spirometra theileri from a Leopard (Panthera pardus) and Spotted Hyena (Crocuta crocuta) in Tanzania

In the present study, a Spirometra species of Tanzania origin obtained from an African leopard (Panthera pardus) and spotted hyena (Crocuta crocuta) was identified based on molecular analysis of cytochrome c oxidase I (cox1) and NADH dehydrogenase subunit I (nad1) as well as by morphological observations of an adult tapeworm. One strobila and several segments of a Spirometra species were obtained from the intestine of an African male leopard (Panthera pardus) and spotted hyena (Crocuta crocuta) in the Maswa Game Reserve of Tanzania. The morphological characteristics of S. theileri observed comprised 3 uterine loops on one side and 4 on the other side of the mid-line, a uterine pore situated posterior to the vagina and alternating irregularly either to the right or left of the latter, and vesicular seminis that were much smaller than other Spirometra species. Sequence differences in the cox1 and nad1 genes between S. theileri (Tanzania origin) and S. erinaceieuropaei were 10.1% (cox1) and 12.0% (nad1), while those of S. decipiens and S. ranarum were 9.6%, 9.8% (cox1) and 13.0%, 12.6% (nad1), respectively. The morphological features of the Tanzania-origin Spirometra specimens coincided with those of S. theileri, and the molecular data was also consistent with that of S. theileri, thereby demonstrating the distribution of S. theileri in Tanzania. This places the leopard (Panthera pardus) and spotted hyena (Crocuta crocuta) as new definitive hosts of this spirometrid tapeworm.

Reverse Warburg Effect-Related Mitochondrial Activity and ¹⁸F-FDG Uptake in Invasive Ductal Carcinoma / 대한핵의학회잡지

PURPOSE: We evaluated the relationship between fluorine-18 fluoro-2-deoxy-glucose (¹⁸F-FDG) uptake and mitochondrial activity in cancer cells and investigated the prognostic implications of this relationship in patients with invasive ductal carcinoma of the breast (IDCB).METHODS: One hundred forty-six patients with primary IDCB who underwent preoperative ¹⁸F-FDG PET/CT followed by curative surgical resection were enrolled in the current study. Mitochondrial activity of cancer cells was assessed based on translocase of outer mitochondrial membrane 20 (TOMM20) expression and cytochrome C oxidase (COX) activity. A Pearson's correlation analysis was used to assess the relationship between the maximum standardized uptake value of the primary tumour (pSUVmax) and mitochondrial activity. Clinicopathological factors, including pSUVmax, histological grade, oestrogen receptor (ER), progesterone receptor (PR), and TOMM20 expression; and COX activity, were assessed for the prediction of disease-free survival (DFS) using the Kaplan–Meier method and Cox proportional hazards model.RESULTS: Fourteen of the 146 subjects (9.6%) showed tumour recurrence. There was a significant positive correlation between ¹⁸F-FDG uptake and the mitochondrial activity of cancer cells in patients with IDCB, and increased ¹⁸F-FDG uptake and mitochondrial activity were significantly associated with a shorter DFS. Additionally, results from the receiver-operating curve analysis demonstrated that the cut-off values of pSUVmax, TOMM20 expression, and COX activity for the prediction of DFS were 7.76, 4, and 5, respectively. Further, results from the univariate analysis revealed that pSUVmax, TOMM20 expression, PR status, and histologic grade were significantly associated with DFS; however, the multivariate analysis revealed that only pSUVmax was associated with DFS (HR, 6.51; 95% CI, 1.91, 22.20; P = 0.003).CONCLUSIONS: The assessment of preoperative ¹⁸F-FDG uptake and post-surgical mitochondrial activity may be used for the prediction of DFS in patients with IDCB.

Cytochrome C Oxidase Subunit 1, Internal Transcribed Spacer 1, Nicotinamide Adenine Dinucleotide Hydrogen Dehydrogenase Subunits 2 and 5 of Clonorchis sinensis Ancient DNA Retrieved from Joseon Dynasty Mummy Specimens

We analyzed Clonorchis sinensis ancient DNA (aDNA) acquired from the specimens of the Joseon mummies. The target regions were cytochrome C oxidase subunit 1 (CO1), internal transcribed spacer 1 (ITS1), nicotinamide adenine dinucleotide hydrogen (NADH) dehydrogenase subunits 2 (NAD2) and 5 (NAD5). The sequences of C. sinensis aDNA was completely or almost identical to modern C. sinensis sequences in GenBank. We also found that ITS1, NAD2 and NAD5 could be good markers for molecular diagnosis between C. sinensis and the other trematode parasite species. The current result could improve our knowledge about genetic history of C. sinensis.

Ten Cases of Taenia saginata Infection Confirmed by Analysis of the Internal Transcribed Spacer 1 rDNA Region in the Republic of Korea

From October 2015 to August 2018, tapeworm proglottids were obtained from 10 patients who were residents of Daegu and Gyeongbuk provinces and had a history of raw beef consumption. Most of them had no overseas travel experience. The gravid proglottids obtained from the 10 cases had 15–20 lateral uterine branches. A part of internal transcribed spacer 1 (ITS1) DNA of the 10 cases, amplified by polymerase chain reaction (PCR) and digested with AleI restriction enzyme, produced the same band pattern of Taenia saginata, which differentiated from T. asiatica and T. solium. Sequences of ITS1 and cytochrome c oxidase subunit 1 (cox1) showed higher homology to T. saginata than to T. asiatica and T. solium. Collectively, these 10 cases were identified as T. saginata human infections. As taeniasis is one of the important parasitic diseases in humans, it is necessary to maintain hygienic conditions during livestock farming to avoid public health concerns.

Morphological and Molecular Identification of Stellantchasmus dermogenysi n. sp. (Digenea: Heterophyidae) in Thailand

We tried a series of morphological and molecular approaches to identify a new species of Stellantchasmus (Digenea: Heterophyidae) originating from the wrestling half-beaked fish, Dermogenys pusillus of Thailand. Adult worm samples of the new species were recovered from hamsters experimentally infected with the metacercariae from D. pusillus in Thailand. Two isolates (Thai and Korean) of Stellantchasmus falcatus were used as comparative control groups. Worm samples of 3 Stellantchasmus groups were morphologically observed and molecularly analyzed with the mitochondrial cytochrome c oxidase 1 gene. The morphological characteristics of S. dermogenysi n. sp. are similar to S. falcatus originating from brackish water fish, but minor difference was noted including the absence of the prepharynx, position of the ovary near the ceca end, smaller body size, and shorter esophageal length. A phylogenetic tree derived from neighbor-joining and maximum-likelihood methods suggests that S. dermogenysi n. sp. is separated from S. falcatus supported by high bootstrap values. The relative divergences persist between these host-specific trematodes, which we suggest should be recognized as 2 distinct species. Comparisons of S. dermogenysi n. sp. with S. falcatus isolated from mullets in Thailand and Korea indicate a genetic divergence of mitochondrial DNA of 19.4% and 21.7%, respectively. By the present study, a new species, Stellantchasmus dermogenysi n. sp. (Digenea: Heterophyidae), is proposed in Thailand based on molecular evidences, in addition to minor morphological differences between S. falcatus and the new species.

Four Taeniasis saginata Cases Diagnosed at a University Hospital in Korea

In recent years, the taeniasis has been rarely reported in the Republic of Korea (Korea). But in this study, we intend to report 4 taeniasis cases caused by Taenia saginata during a 5-month period (February to June 2018) at a unversity hospital in Gwangju, Korea. Worm samples (proglottids) discharged from all cases were identified by phenotypic and molecular diagnostics. Mitochondrial cytochrome c oxidase subunit I sequences showed 99.4–99.9% identity with T. saginata but, differed by 4% from T. asiatica and by 7% from T. multiceps, respectively. We found that tapeworms in 2 cases (Cases 2 and 3) yielded exactly the same sequences between them, which differed from those in Cases 1 and 4, suggesting intra-species variation in tapeworms. These taeniasis cases by T. saginata infection in this study, which occurred within a limited time period and region, suggest the possibility of a mini-outbreak. This study highlights the need for further epidemiological investigation of potentially overlooked cases of T. saginata infection in Korea.

Molecular characterization of Plasmodium juxtanucleare in Thai native fowls based on partial cytochrome C oxidase subunit I gene / 대한수의학회지

Avian malaria is one of the most important general blood parasites of poultry in Southeast Asia. Plasmodium (P.) juxtanucleare causes avian malaria in wild and domestic fowl. This study aimed to identify and characterize the Plasmodium species infecting in Thai native fowl. Blood samples were collected for microscopic examination, followed by detection of the Plasmodium cox I gene by using PCR. Five of the 10 sampled fowl had the desired 588 base pair amplicons. Sequence analysis of the five amplicons indicated that the nucleotide and amino acid sequences were homologous to each other and were closely related (100% identity) to a P. juxtanucleare strain isolated in Japan (AB250415). Furthermore, the phylogenetic tree of the cox I gene showed that the P. juxtanucleare in this study were grouped together and clustered with the Japan strain. The presence of P. juxtanucleare described in this study is the first report of P. juxtanucleare in the Thai native fowl of Thailand.

Analysis of COI and ITS2 regions of DNA obtained from Paragonimus westermani eggs in ancient coprolites on Joseon dynasty mummies

The genetic information of ancient Paragonimus westermani, the oriental lung fluke infecting over 20 million people worldwide, has not been thoroughly investigated thus far. We analysed genetic markers (COI and ITS2) of P. westermani from coprolite specimens (n = 6) obtained from 15th to 18th century Korean mummies. Our results indicated that all P. westermani sequences were generally distinct from the other species of the genus Paragonimus. The sequences were clustered into three groups: Group I for East Asia; Group II for South and Southeast Asia; and Group III for India and Sri Lanka. In this study, we found that ancient P. westermani sequences in Korea belong to Group I, adding invaluable information to the existing knowledge of Paragonimus paleogenetics.

Especies Crípticas Lutzomyia longipalpis (Diptera: Phlebotominae) y sus Implicaciones en la Transmisión de Leishmaniasis en Panamá / Cryptic Species Lutzomy ia longipalpis (Diptera: Phlebotominae) and its Implications in the Transmission of Leishmaniasis in Panama

Lutzomyia longipalpis es el principal v ector de una importante enfermedad desatendida en América. La diversidad genética de este vector se estimó en la población colectada en dos áreas geográficas separadas por hasta 37 km. Analizamos la secuencia CB3­PDR / N1N­PDR de 22 individuos obte­ niendo un parámetro de: h = 0.43 y π = 0.0017 (Bona), h = 0.89, π = 0.004 (El Limón) con una dife­ renciación genética de kst = 0.03; p> 0.05 entre ellos. Ocho haplotipos fueron detectados, de los cuales fue compartido. Se detectó una diferenciación significativa entre las poblaciones Panamá­ Colombia (kst = 0.98), Panamá­Costa Rica (kst = 0.98) y Panamá­Brasil (kst = 0.72) bajo el modelo de aislamiento. Las inferencias genéticas de esta población pueden complementar la información de la capacidad de dispersión y brindar pistas importantes para comprender la ecología de Lutzom­yia longipalpisen Panamá.

Lutzomyia longipalpis is the main vector of an important neglected disease in America. The genetic div ers ity of this vector was estimated in the population collected in two geographical areas separated by up to 37 km. We analyzed the sequence CB3­PDR / N1N­PDR of 22 individuals obtaining a parameter of: h = 0.43 and π = 0.0017 (Bona), h = 0.89, π = 0.004 (The Lemon) with a genetic differentiation of kst = 0.03; p> 0.05 between them. Eight haplotypes were detected, of which it was shared. A significant differentiation was detected between the Panama­Colombia (ks t = 0.98), Panama­Costa Rica (kst = 0.98) and Panama­Brazil (kst = 0.72) populations under the isolation model. The genetic inferences of this population can complement the dispersion information and provide important clues to understand the ecology of Lutzomyia longipalpis in Panama.

The Taxonomic Status of Spermophilus in the Plague Area of Dingbian County, Shaanxi Province, China / 生物医学与环境科学（英文）

This study was conducted to define the taxonomic status of Spermophilus in the plague area of Dingbian County in Shaanxi Province, China, through the two-factor variance analysis of morphological characteristics, DNA barcoding, and chromosome karyotype analysis. The Spermophilus samples collected from Dingbian and Zhengxiang Baiqi Counties exhibited significant differences in their morphological measurements. All Spermophilus samples form two distinct branches in neighbor-joining (NJ) tree. One branch included the Spermophilus samples collected from Inner Mongolia, and the other branch included samples collected from the plague foci of Shaanxi Province and the Ningxia Region. The Spermophilus samples collected from Dingbian County had a chromosome number of 2n = 38 in 84.40% of all their cells. The Spermophilus species collected from the plague area of Dingbian County was categorized as Spermophilus alashanicus (S.alashamicus). The findings reported in this study are epidemiologically significant for monitoring plague in this region of west-central China.

Molecular Characterization of Hard Ticks by Cytochrome c Oxidase Subunit 1 Sequences

Although widely studied, the natural diversity of the hard tick is not well known. In this study, we collected 194 sequences from 67 species, covering 7 genera of hard tick. The 5′ region of the mitochondrial cytochrome c oxidase subunit 1 region (586 bp) has been used to investigate intra- and inter-species variation and the phylogenetic tree of neighbor joining method has been used for assessment. As a result, by comparing the K2P-distance of intra- and interspecies, 30 samples (15.2%) shown that interspecies distance was larger than the minimum interspecfic distance. From the phylogenetic analysis, 86.8% (49) of the species were identified correctly at the genus level. On deeper analysis on these species suggested the possibility of presence cryptic species. Therefore, further work is required to delineate species boundaries and to develop a more complete understanding of hard tick diversity over larger scale.

Molecular Genetic Findings of Spirometra decipiens and S. ranarum in Korea

The taxonomy of Spirometra species has been controversial despite the medical and veterinary importance. Currently, only a few Spirometra species are considered valid species in the genus Spirometra. In the present study, the distribution of Spirometra species obtained from animals in Korea were identified by molecular analysis of the mitochondrial cytochrome c oxidase I (cox1) gene. A total of 28 Spirometra species specimens were analyzed. These were all collected between 1973 and 2008 in the Republic of Korea. Mitochondrial cox1 sequences were examined for a total of 28 specimens comprising 14 S. decipiens and 14 S. ranarum. The difference in partial cox1 sequences (316 bp) between S. erinaceieuropaei (KJ599680) and S. ranarum (this study) was 9.3%, while that between S. decipiens (KJ599679) and S. ranarum (this study) was 2.2%. Genetic analyses identified 2 Spirometra species in animals such as cat, leopard cat, dog, duck and snake in Korea as S. decipiens and S. ranarum. S. decipiens and S. ranarum were present in Gyeongnam Province (P), Jeonnam P, Gangwon P, Chungbuk P, and Seoul. S. decipiens was found in tadpoles, snakes, ducks, cats, leopard cats and dogs, while S. ranarum was found in cats and dogs. The ratio of S. decipiens:S. ranarum calculated from the molecular data was 14:14 (or 1:1). These results indicate that S. decipiens and S. ranarum are sympatrically distributed in Korea.

Especies de flebotomíneos (Diptera: Psychodidae) recolectados en reservas naturales de las regiones del Darién y del Pacífico en Colombia / Species of sand flies (Diptera: Psychodidae) collected from natural reserves in the Pacific and Darien regions of Colombia

Resumen Introducción. Los departamentos de Chocó y Antioquia en Colombia presentan condiciones climáticas y de vegetación que favorecen el establecimiento de especies de vectores del género Lutzomyia y la transmisión de Leishmania spp. a poblaciones humanas que ingresan a ambientes selváticos conservados. Objetivo. Reportar las especies de flebotomíneos presentes en tres reservas naturales de las regiones del Darién y del Pacífico en Colombia. Materiales y métodos. Los flebotomíneos se recolectaron en las reservas naturales El Aguacate (Acandí, Chocó), Nabugá (Bahía Solano, Chocó) y Tulenapa (Carepa, Antioquia). La recolección se hizo con trampas de luz CDC, mediante búsqueda activa en sitios de reposo y con trampas Shannon. La determinación taxonómica de especies se basó en las claves taxonómicas. En algunas especies de interés taxonómico, se evaluó el uso de secuencias parciales de la región 5' del gen COI. Resultados. Se recolectaron 611 flebotomíneos adultos: 531 en Acandí, 45 en Carepa y 35 en Bahía Solano. Se identificaron 17 especies del género Lutzomyia, tres del género Brumptomyia y una del género Warileya. Las distancias genéticas (K2P) y los soportes de agrupación (>99 %) en el dendrograma de neighbor joining correspondieron a la mayoría de unidades taxonómicas operacionales moleculares (Molecular Operational Taxonomic Units, MOTU) establecidas para el grupo Aragaoi y confirmaron claramente la identidad de Lu. coutinhoi. Conclusión. Se identificaron especies que tienen importancia en la transmisión de la leishmaniasis en Acandí, Bahía Solano y Carepa. Se confirmó la presencia de Lu. coutinhoi en Colombia.

Abstract Introduction: The departments of Chocó and Antioquia in Colombia show climatic and vegetation conditions favoring the establishment of vector species of the genus Lutzomyia and the transmission of Leishmania spp. to human populations entering conserved forest environments. Objective: To report the species of Phlebotomine sandflies present in three natural reserves in the Darien and Pacific regions of Colombia. Materials and methods: Sand flies were collected specifically in the natural reserves El Aguacate (Acandí, Chocó), Nabugá (Bahía Solano, Chocó) and Tulenapa (Carepa, Antioquia). Sand flies were collected with CDC light traps, active search in resting places and Shannon traps. The taxonomic determination of species was based on taxonomic keys. For some species of taxonomic interest, we evaluated the partial sequences of the 5' region of COI gene. Results: A total of 611 adult sand flies were collected: 531 in Acandí, 45 in Carepa and 35 in Bahía Solano. Seventeen species of the genus Lutzomyia, three of the genus Brumptomyia and one of the genus Warileya were identified. The genetic distances (K2P) and grouping supported (>99%) in the neighbor joining dendrogram were consistent for most established molecular operational taxonomic units (MOTU) of the Aragaoi group and clearly confirmed the identity of Lu. coutinhoi. Conclusion: Species that have importance in the transmission of leishmaniasis in Acandí, Bahía Solano and Carepa were identified. The presence of Lu. coutinhoi was confirmed and consolidated in Colombia.

Infección de Aedes albopictus (Skuse, 1894) con el genotipo asiático-americano del virus del dengue serotipo 2 en Medellín y su posible papel como vector del dengue en Colombia / Aedes albopictus (Skuse, 1894) infected with the American-Asian genotype of dengue type 2 virus in Medellín suggests its possible role as vector of dengue fever in Colombia

Resumen Introducción. Aedes aegypti y Ae. albopictus son reconocidos vectores de arbovirus como los del dengue, la fiebre amarilla, el chikungunya y el Zika, en regiones tropicales y subtropicales del mundo. En Colombia, la distribución geográfica de Ae. albopictus ha sufrido un incremento y hoy incluye ciudades como Cali y Medellín. Hasta ahora, sin embargo, no se ha recabado información concluyente sobre su infección viral y su capacidad de transmisión a los humanos. Objetivo. Determinar la infección natural por dengue en ejemplares de Ae. albopictus recolectados en un área urbana de Medellín. Materiales y métodos. Se recolectaron individuos de Ae. albopictus en el campus de la Universidad Nacional de Colombia, sede Medellín. Se confirmó su clasificación taxonómica mediante el análisis del gen citocromo oxidasa I (COI), y se extrajo el ARN total para la identificación del virus del dengue y de los respectivos serotipos. La presencia del genotipo DENV se infirió mediante el análisis del gen NS3. Resultados. El análisis del COI corroboró el estatus taxonómico de Ae. albopictus. Uno de los mosquitos procesados fue positivo para DENV-2 y el análisis del NS3 mostró una gran similitud con el genotipo asiático-americano. Conclusión. Se reporta la infección con DENV-2 en Ae. albopictus en Medellín, Colombia. La presencia del genotipo asiático-americano en una zona urbana sugiere su posible circulación entre humanos y en Ae. albopictus, lo cual alerta sobre su eventual papel en la transmisión del DENV-2, y sobre la necesidad de incluir esta especie en la vigilancia entomológica en Colombia.

Abstract Introduction: Aedes aegypti and Ae. albopictus are recognized vectors of dengue, yellow fever, chikungunya and Zika arboviruses in several countries worldwide. In Colombia, Ae. albopictus geographical distribution has increased to include highly populated cities such as Cali and Medellín. Although this species has been frequently found in urban and semi-urban zones in the country, its role as vector of the dengue fever is poorly known. Objective: To identify the presence of Ae. albopictus specimens naturally infected with dengue virus collected in Medellín. Materials and methods: Insects were collected in the Universidad Nacional de Colombia campus in Medellín. Individuals were classified as Ae. albopictus and confirmed by DNA barcode region analysis. Mosquitoes were processed for dengue virus identification, and a fragment of the NS3 gen was sequenced and compared with DENV-2 genotypes reported in the literature. Results: Sequence analysis of COI indicated Ae. albopictus individuals were similar to those recently reported in Colombia, and genetically close to those from other regions worldwide. Among the pools tested one was positive for DENV-2, and the NS3 analysis indicated it belonged to the Asian-American clade. Conclusion: We report the presence Ae. albopictus naturally infected with the Asian-American genotype of DENV-2 in Colombia. The presence of Ae. albopictus specimens carrying the most common genotype infecting humans in a highly populated city such as Medellín indicates its potential role as dengue vector in Colombia and highlights the relevance of including it in current vector surveillance strategies.