Molecular detection of the botanical origin of pollen in honey bee-collected pellets: a comparison of methods

Dupla diplomação com a Université Libre de TunisIdentification of botanical origin of mixed pollen samples has several applications, including assessment of plant-pollinator interactions, botanical origin of honey, monitoring of pesticide use, monitoring of allergy-related airborne pollen sources, among others. Such applications, however, have previously been limited to conventional pollen identification via light microscopy, which usually has low taxonomic resolution and requires expert knowledge. One alternative for botanical identification of mixed pollen samples is to use of DNA metabarcoding high throughput sequencing (HTS), which could overcome these drawbacks. Recent studies demonstrate that the nuclear barcoding marker ITS2 (internal transcribed spacer 2 region of nuclear ribosomal DNA) can be amplified from DNA extracted from mixed pollen samples. The aim of this study was to compare a variety of methods of storage/transportation and DNA extraction that ensure good DNA yield and quality appropriate for botanical identification of mixed pollen samples by means of a DNA metabarcoding approach, combining the amplification of ITS2 with HTS. In the context of the international project “INSIGNIA: environmental monitoring of pesticide use through honeybees”, mixed pollen samples were collected from traps set up in apiaries from several European countries, stored by beekeepers and later transported to the laboratory of CIMO for identification of plant taxa and inference of relative abundances. Four methods of genomic DNA isolation (NucleoSpin Food kit, GF-1 Plant kit, HigherPuritykit, and CTAB- PVP) were compared regarding DNA yield and purity by means of spectrophotometry and standard gel electrophoresis. Additionally, four storage/transportation methods of trap- collected pollen samples (freezing at -20 °C, drying at 25°C for 2 days, drying with silica, and placing in ethanol) were compared to assess their impact on the quality and quantity of extracted DNA. The results demonstrated the superior efficacy of the NucleoSpin DNA extraction method. The different storage/transportation conditions of pollen samples were compared for their impact on DNA quality and quantity using the NucleoSpin as the DNA extraction method. The results showed that the DNA extracted from the pollen samples placed in ethanol had the best quality/yield compared to the DNA extracted from the other samples with different storage conditions. Two primer pairs targeting ITS2 region ITS- S2F/ITS4R and ITS-u3/ITS-u4, were employed to identify plant taxa via metabarcoding HTS. The number of taxa identified in common using these two primers were 48 families, 118 genera, and 204 species, corresponding to 87.2 % , 79.5%, and 68.7%, respectively.The results of identification of taxa we present very similar results, making comparisons difficult, with a slight difference in the number of taxa (ITS-u3/ITS-u4 with higher number of identified taxa) and the abundance (ITS-S2F/ITS4R with higher abundance of taxa identified). This study thus offers improvements in the laboratory workflow ensuring a good DNA quantity and quality for downstream HTS applications.A identificação da origem botânica de amostras de pólen tem várias aplicações, incluindo avaliação das interações planta-polinizador, origem botânica do mel, monitorização do uso de pesticidas, monitorização de fontes de pólen do ar causadoras de alergias, entre outras. Tais aplicações, no entanto, têm sido limitadas pela identificação convencional de pólen por microscopia óptica, que geralmente possui baixa resolução taxonómica e requer conhecimento especializado. Uma alternativa para a identificação botânica de amostras mistas de pólen é o uso de “DNA metabarcoding high throughput sequencing (HTS)”, que pode superar essas desvantagens. Estudos recentes mostram que o código de barras nuclear ITS2 (região espaçadora interna transcrita 2 do DNA ribossómico nuclear) pode ser amplificado a partir de DNA extraído de amostras mistas de pólen. O objetivo deste estudo foi comparar uma variedade de métodos de armazenamento/transporte e extração de DNA que garantam um bom rendimento e qualidade do DNA adequados para a identificação botânica de amostras de pólen misto por meio de uma abordagem de DNA metabarcoding, combinando a amplificação de ITS2 com HTS. No contexto do projeto internacional “INSIGNIA: monitorização ambiental do uso de pesticidas através das abelhas”, foram colhidas amostras de pólen a partir de capta polens montados em apiários em vários países da Europa, armazenadas pelos apicultores e posteriormente foram transportadas para o laboratório do CIMO para identificação botânica e inferência das abundâncias relativas. Quatro métodos de isolamento de DNA genómico (NucleoSpin Food kit, GF-1 Plant kit, HigherPuritykit e CTAB-PVP) foram comparados quanto ao rendimento e pureza do DNA por meio de espectrofotometria e eletroforese em gel de agarose. Adicionalmente, quatro métodos de armazenamento/transporte de amostras de pólen (congelamento a -20 ° C, secagem a 25 ° C por 2 dias, secagem com sílica e colocação em etanol) foram comparados para avaliar o seu impacto na qualidade e quantidade do DNA extraído. Os resultados demonstraram a superioridade do método de extração de DNA NucleoSpin. As diferentes condições de armazenamento/transporte das amostras de pólen foram comparadas quanto ao seu impacto na qualidade e quantidade do DNA, usando o NucleoSpin como método de extração de DNA. Os resultados mostraram que o DNA extraído das amostras de pólen colocadas em etanol apresentou a melhor relação qualidade/rendimento comparado com o DNA extraído das outras amostras submetidas a diferentes condições de armazenamento. Dois pares de primers da região ITS2, ITS- S2F/ITS4R e ITS-u3/ITS-u4, foram utilizados para identificar os taxa representados nas amostras de pólen através do método DNA metabarcoding HTS. O número de taxa identificados em comum usando estes dois pares de primers foi de 48 famílias, 118 géneros e 204 espécies, correspondendo a 87,2%, 79,5% e 68,7%, respectivamente. A análise dos resultados sugere que os dois pares de primers são muito semelhantes, com uma pequena diferença no número de taxa (ITS-u3/ITS-u4 com maior número de taxa identificados) e na abundância (ITS-S2F/ITS4R com maior abundância de taxa identificados). Este estudo contribuiu para a melhoria do fluxo de trabalho laboratorial garantindo uma boa quantidade e qualidade do DNA com vista a aplicações HST a jusante.This work was developed in the framework of the international project “INSIGNIA: environmental monitoring of pesticide use through honeybees”, supported by the Health and Food Safety Directorate General; SANTE/E4/SI2.788418-SI2.788452-INSIGNI

Impact de la gouvernance et de l'audit sur la performance de l'entreprise

Cet article traite de l'impact des mécanismes de gouvernance et de l'audit sur la performance de l'entreprise. Le nombre important de scandales financiers qui ont ébranlés le monde des affaire ces dernières années, un peu partout dans le monde, a conduit à la promulgation de diverses reformes et lois sur la sécurité financière. Suite auxquelles, l'importance de la gouvernance et de l'audit s'est accrue. Nous avons utilisé les modèles de Panel pour l'estimation empirique sur un échantillon correspondant à la population des entreprises tunisiennes cotées sur la BVMT entre 2002 et 2005. Pour l'estimation de la performance, nous avons retenu trois mesures le ROA, le Q de Tobin et le ratio de Marris. Pour appréhender la gouvernance nous avons retenu le conseil d'administration, sa taille, son indépendance et la concentration du capital. Pour l'estimation de la qualité de l'audit, nous avons retenu l'indépendance de l'auditeur et sa compétence.gouvernance; audit; mécanismes de contrôle; performance de l'entreprise

Book Review: Inequality in America: Causes and Consequences

Inequality in America: Causes and Consequences is a recent book authored by Rycroft (economics, Univ. of Mary Washington, Fredericksburg, VA) and Kinsley (business law, Univ. of Mary Washington, Fredericksburg, VA) and was written by scholars, advanced graduate students, and expert practitioners. Their book attempts to bridge the gap in scholarship and practice by incorporating the causes and consequences of history, the current state of inequality in the United States, and future consequences if left unresolved. This book is structured according to fundamental topics such as education, healthcare, housing, environmental issues, and the justice system. Using an effective approach to teaching the reader about the concepts, the book then teaches how to practically apply the concepts in real-world situations for optimal results. While the book lacks specificity in the discussion of a global perspective or the impact of leadership approach styles on inequality through history until now, it does provide comprehensive explanations combined with practical instructions for students and practitioners

Twin deliveries in Sacred Heart Catholic Hospital Obudu: experience from a secondary health facility in South-South Nigeria

Background: Twin pregnancy is associated with increased risk of obstetric complications as well as increased perinatal mortality rate. The study aims to determine the incidence and outcome of twin pregnancy in a rural/semi urban hospital which provide secondary level obstetric services in South- south Nigeria.Method: This was a retrospective review of all twin deliveries at the Sacred Heart Hospital, Obudu.The case files of all patients who had twin deliveries from 1st January 2009 to 31st December 2014 were studied. The data extracted were age, parity; presentation of foetuses, mode of delivery, gestational age at delivery, maternal and foetal complications, Apgar score and foetal outcome. The data was analysed using IBM SPSS version 20 package. A p-value of < 0.05 was considered significant.Results: The incidence of twin pregnancy was 1:32 deliveries. The mean age of the women was 27.0 +- 5.3 years. The highest incidence was among 25-29 years age group.The median parity was 2,range 0-7.About 78.1% (139) of the women were booked, while 21.9% were unbooked.Twin pregnancy was diagnosed before the onset of labour in 73.6% of the women.The mean gestational age at delivery was 37.1 +- 3.4 weeks, with 19.1% (34) delivered preterm.Preterm delivery was the commonest complication (39.9%), followed by retained second twin. Abnormal presentation of the leading twin was the most common indication for Caesarean section (41.9%), followed by retained second twin (12.9%).About 66.8% of the patients had the leading twin in cephalic presentation, cephalic: cephalic presentation was the most common presentation (43.8%).The mean birthweight of the first twin was 2.4+-0.6kg and the second twins 2.3+-0.6kg. The first twin had fewer babies with 5-minute Apgar score less than 7 compared to the second,there were also more stillbirths among the second twins, P< 0.0001.The overall stillbirth rate was 112.7/1000 births compared with 63.1/1000 for all singleton deliveries in the centre.Conclusion: Twin pregnancy is common in our secondary health facility and is associated with high maternal and foetal complications and stillbirth rate. More in-service training of doctors and midwives on skills of twin delivery and early decisions on referral for specialised care are recommended.Keywords: Twin deliveries, secondary health facility, foetal outcom

Impact of the Nigerian Land Use Act on Economic Development in the Country

This paper appraises the Nigerian Land Use Act and its capacity to engender economic development and prosperity. The Land Use Act was enacted in 1978 with the aim of energizing economic development by ensuring effective and equitable utilization of land and land resources in the country. As this paper has found out, however, achieving this lofty objective has been hampered by two major obstacles. The first is the inherent contradictions and defects in the law, and the second is institutional weakness and lack of political will to implement the Act fairly and equitably. The paper has further discovered that the result of these anomalies is a failure of the Act to accomplish some of its major objectives. To re-focus the Act and secure economic development and easy access to land by both government and the citizens, the paper has recommended the amendment of certain provisions of the Act in addition to mustering the political will by government to implement the amended version of the Act in a fair and equitable way.&nbsp

BUSINESS ANALYSIS AND FUNCTIONALITY DESIGN FOR CONNECT COOPERATIVE

This thesis was aimed at establishing what were the business requirements for a small start-up company in the name of Connect Cooperative. The commissioning body of the thesis is HAMK UAS indirectly doing so on behalf of the Connect Consult start-up which itself generated from the Connect project in which students and staffs of HAMK contributed by running various sub researches and information. From this business requirement a set of technical requirements were being generated with a major restriction placed on the project or research scope limiting the project to the soft-component of the system in accordance to Sun Java’s guide on deployment planning clearly stating the importance on placing boundaries so as to avoid infinite scenarios or situations. The search starts by clearly identifying the adopted organization structure of Connect cooperative and presenting and classifying the proposed start-up’s services. The thesis further goes on to articulate how this organization structure and proposed services could be integrated into an online or virtual portal aimed at various purposes but with a major aim of assisting the start-up deliver value and services to its customers. Information gathering was done verbally by attending various meetings with the start-up’s members and also requesting information from the start-up directly such as their previous research results like the one they did in order to identify their human resources. In order to achieve results of standards, guides were being used so as to give a step by step method used in result achievements with a major use of the Sun java’s enterprise deployment plan –white paper guide. The business requirements were identified as the business constrains and a technical requirement with respect to the project from which the use cases were used representing and designing the system's functions