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Balanced medical and criminal justice approaches addressing determinants of alcohol and other drugs (AOD) use are central to achieving the United Nations Sustainable Development Goals 3 and 16. However, data on AOD use in resource-poor settings are characterised by relative paucity and anecdote. This study aimed to describe and analyse AOD use susceptibility and its determinants among school-attending adolescents in the capital Bissau, Guinea-Bissau. Survey data were collected through a locally adapted Planet Youth questionnaire in June 2017 across 16 schools in Bissau. The 2039-strong sample was selected through a multi-stage, random cluster process. Participants’ lifetime alcohol use was 27.3%, smoking 10.8%, and cannabis use 3.6%. In each instance, the ‘15 years and above’ age bracket was the most common initiation period, signifying increased instigation in later adolescence. Subsequent Varimax Principal Component Analysis (PCA) of 312 variables captured data-driven models of AOD use susceptibility, implying imitated, polydrug use among initiates. Linear regression analysis revealed drinking, group violence, male gender, school dismissal and relationship breakdown as explanatory variables. Overlapping predictive variables suggest singular interventions may pre-empt myriad antisocial behaviours. PCA offers alternatives to singular, potentially myopic quantifications of drug use. PCA facilitates the creation of context-sensitive composite variables, enabling the identification of related outcome behaviours relevant to studies’ sociocultural settings.

Digital technology plays an important role in achieving many of the Sustainable Development Goals. However, access is uneven, with 80% of those in high-income countries being online compared to 20% of those in the 47 least developed countries. This study aimed to describe and analyse adolescents&rsquo, access to and usage of digital technology in Guinea-Bissau and its implications. In June 2017, a survey with a locally adapted Planet Youth questionnaire was implemented in the capital, Bissau, whereby classes in 16 secondary schools were surveyed on a variety of issues. In total, 2039 randomly selected students participated, the survey included ten questions specifically on the access to and use of digital technology. Half of the respondents had access to desktop/laptops, and one-third used mobile internet daily, about two-thirds had an experience of social media. Explanatory variables included educational institution, parental education, economic situation, and gender. Furthermore, students&rsquo, experience of social media was significantly linked to bullying, anxiety, depression, smoking and alcohol consumption. Many adolescents in Bissau have no experience of using digital technology, including for schoolwork. Access improvements are necessary so that young Bissau-Guineans are not to be left behind in developing their capabilities and can benefit from proficiency in the use of digital technologies. At the same time, potential harmful usage of the media requires the implementation of preventive measures.

Cashew (

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Cashew (Anacardium occidentale L.) is a cash crop with a highly significant economic importance in West Africa, particularly in Guinea-Bissau (Monteiro et al. 2015, 2017). In October 2018, dieback-like symptoms such as wilt and necrosis of apical shoots were observed in 10% of the cashew trees grown in a 100-plant orchard in Bolama Island at Bijagos archipelago, Guinea-Bissau. Six symptomatic apical shoots from individual plants were collected for fungal isolation and identification. Tissue pieces (3 × 2 mm) from healthy to diseased margins were surface sterilized with 1% sodium hypochlorite, washed twice with sterilized water, placed on potato dextrose agar (PDA, Difco Laboratories) supplemented with potassium thiocyanate (50 µg/ml), and incubated at 24 ± 1°C in the dark for 7 days. Four fungal colonies were isolated (67%) and purified through hyphal tip removal, displaying rapid growth rate, and aerial mycelia that initially was white, turning later to dark greenish on PDA. Pycnidia produced on 1.5% water agar with sterilized pine needles (∼25°C; near-UV light) were solitary, covered by mycelium, obpyriform to ampulliform (152.5 ± 41.6 × 135.2 ± 30.8 μm, n = 30). Conidia were unicellular, hyaline, smooth, fusoid to ovoid, thin-walled, measuring 16.21 ± 1.52 × 5.84 ± 0.66 μm (n = 50, length/width ratio = 2.8). Such morphological features are characteristic of Neofusicoccum spp. (Phillips et al. 2013). For molecular identification, genomic DNA was extracted from a representative isolate (GB160), and partial regions of ribosomal internal transcriber spacer (ITS) (ITS1/ITS4; White et al. 1990), translation elongation factor 1-α (EF1-α) (EF1-688F/EF1-1251R; Alves et al. 2008), and β-tubulin (β-tub) (Bt2a/Bt2b; Glass and Donaldson 1995) genes were amplified as previously described, respectively, with bovine serum albumin (50 mg/ml). Amplicons were sequenced and deposited in GenBank (ITS, MN952993; EF1-α, MN952204; β-tub, MN952208). BLAST analysis of ITS, EF1-α, and β-tub gene sequences showed 100% identity with Neofusicoccum batangarum reference strain CBS124923 (FJ900608, FJ900654, and FJ900635, respectively). Maximum-likelihood and Bayesian inference analyses from the concatenated dataset placed GB160 isolate within the N. batangarum cluster (bootstrap support value = 72%; Bayesian posterior probability = 0.95). For pathogenicity assessment, 3-month-old cashew ‘Caju di Terra’ plants (n = 8) grown in a greenhouse under controlled conditions were inoculated following a randomized block design as described by Lima et al. (2013). Briefly, 3-mm-diameter stem tissue bark was removed and replaced with a 3-mm-diameter PDA plug retrieved from the colony margin. Inoculation wound was covered with sterilized wet cotton and sealed with Parafilm. Eight control plants were only treated with PDA plugs and the wound covered and sealed as described. After 15 days, all inoculated plants displayed similar symptoms to those observed in the field and vascular lesions (10.8 ± 4.0 cm), whereas control plants remained symptomless. Koch’s postulates were fulfilled by successful reisolation of the pathogen from all inoculated stems and identification by morphology and gene sequencing. N. batangarum was identified associated with Anacardium spp. in Brazil (Netto et al. 2017) and recently reported as causing grapevine dieback in Brazil (Rego et al. 2020). To our knowledge, this is the first report of N. batangarum causing cashew dieback in Guinea-Bissau and in West Africa. Occurrence of this disease may represent a significant impact for cashew production because this crop is the major agricultural commodity in Guinea-Bissau.

Inequality in access to quality education is wide-ranging across and within countries. This study examines determinants for adolescents’ school enrolment in private and public schools in Bissau, Guinea-Bissau, including 2,039 randomly selected school-attending adolescents. The survey methodology was built on the Planet Youth collaboration and is the first of its kind in the setting. The critical determinant for attending a private school rather than a public one reflects parental socioeconomic background, i.e., education, employment status, and financial resources, not gender. Overage class enrolment is common, particularly in public schools and among boys rather than girls. National and international collective action and innovative education policies are needed for Bissau-Guinean adolescents to enjoy their right to the quality education they are entitled to.

Since 2013, there have been reports of gummosis on cashew (Anacardium occidentale L.) in Guinea-Bissau; however, identification of the causal agent remains to be addressed (Monteiro et al. 2015, 2017). During harvest season in May 2018, cashew trunks with dark cankers, cracks, and resin-like gum were observed in orchards located in Bolama Island of Bijagos archipelago and in the continental Tombali region of Guinea-Bissau. Samples from healthy to diseased margins of trunk lesions were collected, surface disinfected with 7% sodium hypochlorite, washed twice with sterilized water, placed on potato dextrose agar (DIFCO, Sparks, MD) supplemented with potassium thiocyanate (50 µg/ml, Sigma, Alges, Portugal), and incubated at 24 ± 1°C. Fungi with mycelium immersed and superficial, first white and later dark greenish to grayish black, were consistently isolated after 7 days, developing pycnidia with conidia initially hyaline, later becoming one-septate, dark brown with longitudinal striations and thick-walled. These morphological characteristics fit the Lasiodiplodia species description (Phillips et al. 2013). Pure cultures from two isolates from Bolama (GB78 and GB134) and one from Tombali (GB32) were used for molecular identification. Genomic DNA was extracted from mycelia using a Qiagen Plant DNA Kit (Qiagen), and partial regions of nuclear ITS rDNA (ITS1F, Gardes and Bruns 1993; ITS4R, White et al. 1990), translation elongation factor 1-α (TEF1- α, Alves et al. 2008), and β-tubulin (β-tub, Glass and Donaldson 1995) genes were amplified following previously described conditions (Coutinho et al. 2017) modified with the addition of bovine serum albumin (BSA, 50 mg/ml). Sequences generated were deposited in GenBank under the accession numbers MN952990 to MN952992 for ITS, MN952201 to MN952203 for β-tub, and MN952205 to MN952207 for TEF1-α. Phylogenetic analyses from the combined dataset using maximum-likelihood and Bayesian inference methods placed each isolate in well-supported clades, namely GB78 as L. theobromae, GB32 as L. pseudotheobromae, and GB134 as L. caatinguensis. Pathogenicity tests were performed on 3-month-old cashew plants (eight plants per isolate). Inoculation followed the method of Lima et al. (2013), consisting of excising a 3-mm-diameter tissue bark out and replacing it with a 3-mm-diameter PDA plug removed from the colony margin. The inoculation wound was covered with sterilized wet cotton and sealed with parafilm. Plants inoculated only with PDA plugs with the wound covered and sealed as previously described were used as controls. After 15 days, all inoculated plants displayed similar symptoms to those observed in the field, whereas control plants remained symptomless. The pathogens were successfully reisolated from symptomatic stems, fulfilling Koch’s postulates. Lasiodiplodia spp. have been described as a causal agent of cashew gummosis, an important disease impacting cashew production in northeastern Brazil (Coutinho et al. 2017). Here we report for the first time three Lasiodiplodia species (L. theobromae, L. pseudotheobromae, and L. caatinguensis) as causal agents of cashew gummosis in Guinea-Bissau, where this crop is the most important agricultural commodity, and given its high economic value, a potential reduction in production would negatively impact smallholder livelihoods and annual revenues.

[EN] Background: Phlomis plants are a source of biological active substances with potential applications in the control of phytopathogens. Phlomis purpurea (Lamiaceae) is autochthonous of southern Iberian Peninsula and Morocco and was found to be resistant to Phytophthora cinnamomi. Phlomis purpurea has revealed antagonistic effect in the rhizosphere of Quercus suber and Q. ilex against P. cinnamomi. Phlomis purpurea roots produce bioactive compounds exhibiting antitumor and anti-Phytophthora activities with potential to protect susceptible plants. Although these important capacities of P. purpurea have been demonstrated, there is no transcriptomic or genomic information available in public databases that could bring insights on the genes underlying this anti-oomycete activity. Results: Using Illumina technology we obtained a de novo assembly of P. purpurea transcriptome and differential transcript abundance to identify putative defence related genes in challenged versus non-challenged plants. A total of 1,272,600,000 reads from 18 cDNA libraries were merged and assembled into 215,739 transcript contigs. BLASTX alignment to Nr NCBI database identified 124,386 unique annotated transcripts (57.7%) with significant hits. Functional annotation identified 83,550 out of 124,386 unique transcripts, which were mapped to 141 pathways. 39% of unigenes were assigned GO terms. Their functions cover biological processes, cellular component and molecular functions. Genes associated with response to stimuli, cellular and primary metabolic processes, catalytic and transporter functions were among those identified. Differential transcript abundance analysis using DESeq revealed significant differences among libraries depending on post-challenge times. Comparative cyto-histological studies of P. purpurea roots challenged with P. cinnamomi zoospores and controls revealed specific morphological features (exodermal strips and epi-cuticular layer), that may provide a constitutive efficient barrier against pathogen penetration. Genes involved in cutin biosynthesis and in exodermal Casparian strips formation were up-regulated. Conclusions: The de novo assembly of transcriptome using short reads for a non-model plant, P. purpurea, revealed many unique transcripts useful for further gene expression, biological function, genomics and functional genomics studies. The data presented suggest a combination of a constitutive resistance and an increased transcriptional response from P. purpurea when challenged with the pathogen. This knowledge opens new perspectives for the understanding of defence responses underlying pathogenic oomycete/plant interaction upon challenge with P. cinnamomi., This work was supported by the project PTDC/AGR-CFL/100217/2008 and the grant SFRH/BD/66016/2009, both funded by Fundacao para a Ciencia e Tecnologia (FCT). The APC charge was supported by the project UID/BIA/04325/2013 - MEDTBIO (FCT). The COST Office and the European Council provided financial support to the Short Term Scientific Mission (COST-STSM-FP0801-10084).

Homology of annotated Phlomis purpurea unigenes to genes from plants. (DOCX 144 kb)

Frequency distribution of lengths of transcript contigs resulting from Illumina HiSeq™ 2000 sequencing. (DOCX 61 kb)

Gene ontology assignments for Phlomis purpurea transcripts. Distribution of Phlomis purpurea contigs into functional sub-categories of Gene Ontology (GO). (DOCX 4917 kb)

Statistics of histological measurements. (DOCX 15 kb)

GC content analysis of Phlomis purpurea transcripts. The average of GC content for P. purpurea was calculated to evaluate the percentage of transcripts with GC content within a range. (DOCX 54 kb)

In order to contribute to the knowledge on the genes involved in the early steps of ectomycorrhiza development, the transcriptional response of Castanea sativa (European chestnut) during the initial contact (6 and 12 h) with Pisolithus tinctorius was analysed by microarray. This study revealed that among the regulated plant genes, a substantial number of up-regulated transcripts showed homology with genes encoding for proteins involved in stress and defense responses, (a cystatin, a cystatin-like protein, a defensin and a universal stress protein). Early contact with the fungal mycelium also altered expression of genes that are putatively involved in cellular processes like signal transduction and communication (receptor kinase-related protein), protein fate (papain-like cystein proteinase), and water transport facilitation (water channel MipK protein). Expression profiling of the early contact between C. sativa and P. tinctorius revealed that changes in gene expression occur few hours after contact, long before the development of a functional mycorrhiza. The induction of genes involved in stress and defense suggests that the host plant reacts rapidly to the presence of the mycobiont eliciting a defense programme similar to that described for pathogenic interactions. Another plant response was the repression of genes normally implicated in water stress accounting for a water stress relief due to the initial contact with the ectomycorrhizal fungus.

In order to elucidate the molecular events associated with pear (Pyrus communis L. cv. Rocha) fruit development and climacteric ripening, high-density cDNA microarrays were constructed. cDNA clones (1364 total) were isolated from a fruit cDNA library and from a subtractive library and arrayed. The expression of these ESTs was monitored from early fruit development, through ripening, until complete fruit senescence, allowing a global coverage of the entire fruit life. A comparison was made with fruit that failed to ripe (FR) due to precocious harvesting. Based on the similarities in transcript expression profiles ESTs were grouped in different clusters. Transcripts encoding kinases and phosphatases were induced specifically during early developmental stages of pear fruit. Another set of genes was activated at the onset of the climacteric period when fruit softening rates also increased. Among these were transcripts encoding for cell wall modifications and pigment and aroma biosynthesis. Transcripts putatively involved in defence response, oxidative stress, primary and secondary metabolism, signalling and transcription regulation were also isolated. According to our results main changes in expression profiles occur in two phases: the cessation of growth and entrance in the initial maturation stages (pre-climacteric), and the entrance into the climacteric period.

Transcriptional changes in Pisolithus tinctorius leading to ectomycorrhizal formation in P. tinctorius– Castanea sativa were investigated using a 12-h fungal interaction in vitro system. Using a 3107-cDNA clone microarray, 34 unique expressed sequence tags (ESTs) were found to be differentially expressed. These ESTs represent 14 known genes, 5 upregulated and 9 downregulated, and 20 orphan sequences. Some transcripts of upregulated genes (with unknown function) were previously identified in other mycorrhizal Pisolithus spp. associations. ESTs for S-adenosyl-l-homocysteine hydrolase and several orphan sequences were identified in our system. The identified transcript of downregulated genes involved hydrophobins, 5S, 18S, and 28S ribosomal RNA genes, large subunits of ribosomal RNA (mitochondrial gene), and two types of heat shock proteins. This study demonstrates the high complexity of molecular events involved in the preinfection steps and suggests the utilization of different fungal gene repertories before ectomycorrhizal formation. These data constitute a first contribution for the molecular understanding of early signaling events between P. tinctorius and C. sativa roots during ectomycorrhizal formation.

A PCR-based method for the synthesis of high quality cDNA is described. This method combines the reverse transcriptase template switching with the assisted recombination cloning. It enables 5′ end enrichment, cDNA synthesis from limiting starting material, elimination of digestion and subcloning steps and substantial time and cost savings.

GC content distribution of Phlomis purpurea transcripts. (DOCX 13 kb)