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Multiple primary malignancies (MPMs) are defined as two or more histopathologically distinct malignancies in the same individual. MPMs are classified as synchronous when tumors are diagnosed within six months of each other. The most common malignancies in MPMs are melanoma, breast, lung, and prostate cancer. Synchronous lymphoma and solid tumors are relatively rare. In these cases, a multi-disciplinary approach to treatment is essential. The early detection of additional primary malignancies such as myeloid and lymphatic tumors will enable prompt management with curative intent. The authors present a case of diffuse B-cell non-Hodgkin lymphoma and invasive lobular breast carcinoma presented as a chylous pleural effusion.

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Crop producers are under great pressure to produce more and better food items. Effective control of crop pathogens is fundamental to guaranteeing food safety and reducing economic losses. Therefore, their identification throughout the production chain is of utmost interest. To achieve this goal, genomic analysis tools are currently being developed allowing to control crop production more effectively.Genomic analysis in some samples is difficult, mostly due to the sample's intrinsic characteristics, i.e., high levels of phenols, fatty acids (e.g., oleaginous fruits, such as olives), and carbon hydrates (e.g., honey), among others. Additionally, some samples yield very low DNA recovery with high content of contaminants, imposing protocol improvements to overcome these difficulties.Here we present protocols focused on qPCR and HRM to detect the presence of fungal pathogens collected from plant-derived samples.

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Grapevine is regarded as a highly profitable culture, being well spread worldwide and mostly directed to the wine-producing industry. Practices to maintain the vineyard in healthy conditions are tenuous and are exacerbated due to abiotic and biotic stresses, where fungal grapevine trunk diseases (GTDs) play a major role. The abolishment of chemical treatments and the intensification of several management practices led to an uprise in GTD outbreaks. Symptomatology of GTDs is very similar among diseases, leading to underdevelopment of the vines and death in extreme scenarios. Disease progression is widely affected by biotic and abiotic factors, and the prevalence of the pathogens varies with country and region. In this review, the state-of-the-art regarding identification and detection of GTDs is vastly analyzed. Methods and protocols used for the identification of GTDs, which are currently rather limited, are highlighted. The main conclusion is the utter need for the development of new technologies to easily and precisely detect the presence of the pathogens related to GTDs, allowing to readily take phytosanitary measures and/or proceed to plant removal in order to establish better vineyard management practices. Moreover, new practices and methods of detection, identification, and quantification of infectious material would allow imposing greater control on nurseries and plant exportation, limiting the movement of infected vines and thus avoiding the propagation of fungal inoculum throughout wine regions.

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The year 2020 has been celebrated as the International Year of Plant Health by the United Nations, and it has been a unique opportunity to realise the vital role of producing while preserving our natural and cultural heritage-Sustainable Food and Agriculture [...].

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Olive anthracnose is caused by fungal species within the Colletotrichum acutatum, C. gloeosporioides and C. boninense complexes. Anthracnose causes severe pre- and post-harvest olive drupe fall. This study aimed to design a species-specific qPCR assay, based on klap1 gene, suitable for C. acutatum s.s. quantification in cv. Galega Vulgar fruit samples. The developed qPCR assay presented a detection limit of 10.14 fg/reaction, and a linear cycle threshold of R2 = 0.996. C. acutatum inoculum was detected in pulverized olive fruits, and in early infection stages, before symptom appearance, 16 h after inoculation (Ct values = 28.29 ± 1.1). In olive samples, the derived melting curve was specific presenting a single dissociation peak (Tmelting = 88.7 °C). The designed assay was effectively applied in C. acutatum detection and quantification using infected olive samples, with a LOD of 0.59 ng and a LOQ of 1.8 ng, allowing its application to orchard management.

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The detection of latent Colletotrichum spp infection in olive drupes is crucial, to avoid contamination in the olive oil production chain. In order to detect the presence of C. acutatum in complex olive matrices a real-time PCR assay was developed, using olive drupe and oil samples from C. acutatum susceptible and tolerant olive cultivars (Galega Vulgar, Cobrançosa and Picual) with different infection levels. A C. acutatum specific sequence, belonging to the Internal Transcribed Spacers region, was used to design the real-time PCR detection assay, resulting in an 490 bp amplicon with a consistent melting temperature (Tm = 87.8 °C). The assay allowed a rapid and high-sensitive C. acutatum detection mean, being able to detect the infection in a latent phase, for the first time, in olive drupes, 16 hai, and in olive oils containing 20% of infected olives. This novel method can be used to monitor C. acutatum presence in olive orchards.

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