Is there a relationship between ferric-chelate reductase activity in roots of poncirus trifoliata and leaf chlorophyll contents?

Poncirus trifoliata is a citrus rootstock very sensitive to Fe deficiency. This deficiency is very common in crops grown in calcareous soils due to the detrimental effect of bicarbonate ion. Higher plants have distinct behaviours when faced with Fe chlorosis, and several mechanisms may be activated under Fe shortage. The aim of this study was to investigate the activity of ferric-chelate reductase (FC-R), a key enzyme in Fe uptake, and to verify whether relationships with leaf chlorophyll contents could be established. Plants were grown in nutrient solutions without Fe (0 μM Fe), with 1 μM Fe, with 120 μM Fe and with 120 μM Fe plus CaCO3 (1 g L-1). Total leaf chlorophyll in young and mature leaves was determined using a calibration model based on a relationship between SPAD readings and concentration of chlorophyll (r2=0.95; P<0.01). The activity of FC-R was determined in roots apexes and several biomass parameters in shoots (number of leaves, height, dry and fresh weight) and roots (dry and fresh weight) were evaluated at the end of the experiment. The activity of FC-R increased in plants grown without iron (0 μM Fe). The results about the relations between root FC-R and leaf chlorophyll are discussed.info:eu-repo/semantics/publishedVersio

Equine sarcoids : a therapeutic challenge

Dissertação de Mestrado Integrado em Medicina VeterináriaEquine Sarcoid is the most common skin tumor in horses and other equidea, where bovine papilomavirus (BPV) plays a major role in its etiopathogenesis. Sarcoids are characterized as unique locally invasive, non-metastatic, fibroblastic skin tumors with unpredictable behavior and are considered to cause substantial economic loss to the equine industry. Several methods have been employed in the treatment of this condition, however, no current approach was found to be 100% effective or universal for all tumor types and locations and recurrence after treatment is common. Therefore, successful management remains a challenge. The aim of this thesis is to identify, through a literature review, the most commonly employed treatments for equine sarcoids, to assess the efficacy of recent treatments observed by the author during the practice period in two different European countries, relating them with other methods and to perform an opinion study through an on-line enquire to Portuguese veterinarians in order to have an idea on how sarcoids are managed in this country. Fifteen horses with a total of 64 tumors were enrolled, from which 29 lesions were treated using 5 different therapeutical approaches: imiquimod, bloodroot (Sanguinaria canadensis) extract, imiquimod and surgical excision, ligation and, laser surgery. Cases treated with imiquimod (AldaraTM), and the bloodroot extract (XxterraTM) had an excellent outcome, with 100% of success rate up to 30 months after finishing treatment, for all locations, dimensions, and types of tumors, with Xxterra being approximately 3 times faster to treat when compared to Aldara. Moreover, these 2 methods are simple, safe, non-invasive, and quite inexpensive when compared with other methods. Disadvantages include long duration of treatment, therefore if quicker results are wished, prior surgical excision followed by application of either ointment could be an alternative. Results suggest that these modalities may be a therapy of choice for sarcoids, although more detailed studies are required to corroborate these findings. No conclusion concerning treatment efficacy could be made for tumors treated by ligation or laser surgery. The opinion study about sarcoid gave an idea of what is performed in Portugal and showed that there are no major differences between the management of this disease in Portugal when compared to other countries.RESUMO - SARCOIDES EQUINO: UM DESAFIO TERAPÊUTICO Sarcoides equino é o tumor cutâneo mais comum em cavalos e outros equidea, onde o papilomavirus bovino (BPV) desempenha um importante papel na sua etiopatogenese. Os sarcoides são caracterizados como tumores cutâneos de fibroblastos, ímpares, localmente invasivos mas não metastizaveis e de comportamento imprevisivel, capazes de causar perdas económicas significativas à indústria equina. Vários métodos têm sido aplicados no tratamento desta condição, no entanto, não existe nenhuma abordagem terapêutica 100% eficaz ou universal para todos os possíveis tipos e localizações do tumor e a recorrência após tratamento é comum. Assim sendo, o sucesso da abordagem terapêutica permanece um desafio. Os objectivos desta tese foram identificar, por meio de revisão bibliográfica, os tratamentos mais frequemente utilizados para o sarcoides equino, avaliar a eficácia de tratamentos recentes observados pelo autor durante o seu período de estágio em dois paises Europeus, relacionando-os com outros métodos e realizar um estudo de opinião, através de um inquérito on-line, a veterinários Portugueses de modo a ter uma noção de como os sarcoides são tratados neste país. Neste trabalho, foram incluídos 15 cavalos com um total de 64 tumores, de entre os quais 29 foram tratados utilizando 5 abordagens terapêuticas distintas: imiquimod, extracto de sanguinária (Sanguinaria canadensis), imiquimod e excisão cirúrgica, laqueação e, cirurgia laser. Os casos tratados com imiquimod (AldaraTM) e com o extracto de sanguinária (XxterraTM), apresentaram um excelente resultado, com taxa de sucesso de 100% até 30 meses após o tratamento, para todas as localizações, dimensões e tipos de tumor, sendo o tempo de tratamento com XxterraTM cerca de 3 vezes mais curto do que com AldaraTM. Estes 2 métodos são simples, seguros, não-invasivos e relativamente baratos quando comparados com outras metodologias. As desvantagens incluem a longa duração do tratamento, no entanto se forem desejados resultados mais rápidos, excisão cirúrgica seguida de aplicação de um dos cremes pode ser uma alternativa. Os resultados sugerem que estas modalidades podem ser uma terapêutica de escolha para sarcoides, no entanto estudos mais detalhados são necessários para corroborar estes resultados. Nenhuma conclusão foi retirada quanto à eficácia de tratamento aplicando extracção por laqueação ou cirurgia laser. O estudo de opinião sobre sarcoides deu uma noção sobre como esta doença é tratada em Portugal revelando não existirem diferenças significativas em relação a outros países

Comparing two in-house developed SNP assays for inferring population structure in the honey bee (Apis mellifera L.)

The honey bee, Apis mellifera L., is under pressure globally due to several factors, one of them is the large-scale introduction of foreign queens and/or colonies which act as vectors of pathogens, and also threaten the genetic integrity of native populations. Different molecular tools have been developed to monitor the genetic integrity of the populations. SNPs (Single Nucleotide Polymorphism) have been preferred because are easily transferred between laboratories, have a low genotyping error, provide high-quality data, and are suitable for automation. Here, we compared the genotyping results obtained with two medium-density-SNP assays previously developed. One of assays was designed from 88 whole genomes of Apis mellifera iberiensis and 44 C-lineage individuals (the main ancestry of commercial bees) using fixed SNPs (FST=1) distributed in the 16 honey bee chromosomes. The other assay was designed from variation in immune genes using a discovery panel of 123 whole genomes, representing seven subspecies (A. m. iberiensis, A. m. mellifera, A. m. intermissa, A. m. sahariensis, A. m. ligustica, A. m. carnica, A. m. siciliana and three lineages (A, M and C). All the samples are from the native range of each subspecies and they were taken from inside the hives, placed in absolute ethanol and stored at -20ºC until DNA extraction. The tools were compared using 473 samples from the Azores, which harbour a genetically complex honey bee population. The samples were genotyped using the iPLEX MassARRAY® MALDI-TOF system. The membership proportions of each individual (Qvalue) were calculated using ADMIXTURE considering two genetic groups (K=2), with 10,000 iterations in 20 independent runs. Our results show that both assays provide similar Q-values, with a Pearson’s correlation of 0.89. Only 9.5% of the samples have an absolute Q-value difference > 0.10. The choice of the best SNP assay depends on the subspecies and the aim of the project. While the immune assays can be applied in different subspecies the other assay was specifically designed for A. m. iberiensis. Furthermore, if there is disease data available, the immune assay caninfo:eu-repo/semantics/publishedVersio

Lasting Developmental Effects of Neonatal Fentanyl Exposure in Preweanling Rats

The present study aimed to determine whether neonatal treatment with fentanyl has lasting effects on stressed developing brain. Six-day-old rats were assigned to one of three groups (10 males/group): (1) fentanyl (incision+fentanyl), (2) saline (incision+0.9% saline), and (3) unoperated (unoperated sham). Pups with a plantar paw incision received repetitive subcutaneous injections of fentanyl or vehicle through postnatal days (PNDs) 6 to 8. A nonoperated sham group served as nonstressed control. Studies included assessment of development from PND 6 to PND 21 (growth indices and behavioral testing). Fentanyl administered twice daily for three days after surgical incision had no impact on early growth and development, as measured on PND 9, but showed a lasting impact on later growth, enhanced behavioral development, and lower anxiety, as measured through PNDs 10–21. While this does not completely support a benefit from such treatment, our findings may contribute to support the neonatal use of fentanyl, when indicated, even in premature newborns

Mitochondrial SNP markers to monitor evolutionary lineage ancestry in Apis mellifera mellifera conservation programs

The European dark honey bee, Apis mellifera mellifera , is threatened inmost of its native range, in part, due to introgressive hybridization with bees from the highly divergent C-lineage, mainly Apis mellifera carnica and Apis mellifera ligustica (De la Rúa et al. 2009; Pinto et al. 2014). Yet, the maintenance of locally adapted genetic diversity is critical for the population long-term survival and sustainability (De la Rúa et al. 2009; Meixner 2010). The growing awareness that genetic diversity is important for sustainable beekeeping led to implementation of different conservation and breeding programs throughout Europe, which are in need of reliable and costefficient molecular tools to accurately monitor Clineage introgression into A. m. mellifera (De la Rúa et al. 2009; Henriques et al. 2018a, b; Meixner 2010). The large mating flight distances and the polyandrous mating system make it challenging to preserve honey bee subspecies in an open conservation area where intruders can fly in (Neumann et al. 1999). It is therefore necessary to regularly control the genetic ancestry of new or superseded colonies.This work was financed by FEDER (Fundo Europeu de Desenvolvimento Regional) through the program COMPETE 2020–POCI (Programa Operacional para a Competitividade e Internacionalização) and by Portuguese funds through FCT (Fundação para a Ciência e a Tecnologia) in the framework of the project BeeHappy (POCI-01-0145-FEDER-029871). Melanie Parejo was supported by a mobility fellowship awarded from the Swiss National Science Foundation (SNSF).info:eu-repo/semantics/publishedVersio

Anestesia Pediátrica em Período de Imaturidade Cerebral e Perturbações do Neurodesenvolvimento

INTRODUCTION: Several experimental and clinical studies suggest that drugs used in pediatric anesthesia may exert undesirable effects on the developing central nervous system. The objective of this review was to assess the results and conclusions of published studies on long lasting neurodevelopment disorders following exposure to anesthetics in children in a phase of brain immaturity. MATERIAL AND METHODS: We performed a literature search in several sources (PubMed, SciELO and Cochrane Library) using the terms 'Pediatric anesthesia OR Pediatric anesthetic OR Developing brain anesthetic OR Developing brain anesthesia AND behavior disorders'. We selected human studies, referring to long lasting neurodevelopment effects after exposure to anesthetics in the first four years of life. RESULTS: Ten retrospective studies met the inclusion criteria. Of these, seven suggest risk of neurobehavioral disorders after exposure of small children to anesthetics, as opposed to the results obtained by the other three. DISCUSSION: Although mostly using large databases, the studies found are retrospective, vary in test groups, include sometimes avoidable confounders and some present inaccuracies in the choice of the test and control populations that can compromise the reliability of the results. CONCLUSION: Because of the numerous limitations of the few studies available, the reported results are still deemed insufficient to change current clinical practice. However, although it is undisputable that anesthesia should be provided when needed, regardless of age, the warnings found in literature are worrisome, therefore whenever surgery is unavoidable in small children, alternatives that may help reduce the risks of anesthetic exposure should be sought.info:eu-repo/semantics/publishedVersio

Development of a new approach based on NADH dehydrogenase subunit 1(ND1) marker and high resolution melting (HRM) analysis towards the authentication of the geographical origin of honey

In the present study, an approach for verifying the geographical origin of honey based on its entomological origin is proposed. The method was developed based on the polymerase chain reaction (PCR) amplification of a new markercontaining different single nucleotide polymorphisms characteristic of honeybees of different mitochondrial (mtDNA) lineages, therefore generating different fluorescent curves on HRM analysis. The method was successfully applied to honeys from Portugal and Italy, expected to be from lineages A and C, respectively, demonstrating their origin compliance.The authors are grateful to Programa Nacional Apícola 2020-2022 for funding the project “AUTENT+ Desenvolvimento de abordagens inovadoras com vista à valorização e exploração do potencial de mercado do mel Português”, to the Foundation for Science and Technology (FCT, Portugal) for financial support by national funds FCT/MCTES to CIMO UIDB/00690/2020, to Fenapícola and Capemel for supplying the Portuguese honeys and to Dr. Antonio Nanetti (CREA-AA) for the Italian honeys. D. Henriques is supported by the project Bee Happy (POCI-01-0145-FEDER-029871) funded by FEDER, COMPETE 2020-POCIand FCT and A.R. Lopes by the PhD scholarship funded by the FCT SFRH/BD/143627/2019.info:eu-repo/semantics/publishedVersio

Colonization patterns of Nosema ceranae in the Azores archipelago

Nosema ceranae and Nosema apis are pathogens of honey bees that cause nosemosis, a disease implicated in colony losses worldwide. They are obligate intracellular pathogens infecting the midgut epithelial cells of adult honey bees. Although N. ceranae was originally a pathogen specific of Apis cerana, currently, it is found in Apis mellifera throughout most of the world. Due to their confined environments, which limited pathogen transmission and dissemination, islands are unique places for epidemiological studies. There are only a few Varroa destructor-free and possibly even fewer N. ceranae-free honey bee sanctuaries in the world, with the Azores being one. Even though with the exception of Santa Maria and Flores, nosemosis has been present in the Azores since 2008, the causal species has yet to be identified. Hence, this study aimed to determine the prevalence and infection levels of Nosema spp. in the Azorean honey bees. In 2014/2015, 474 colonies were sampled on Faial, Flores, Pico, Graciosa, São Jorge, São Miguel, Santa Maria, and Terceira. Additionally, São Jorge, Santa Maria, Faial, and Terceira were re-sampled in 2020 with a total of 91 colonies. DNA was extracted, and the diagnosis and Nosema spp. loads were obtained by multiplex PCR and RT-qPCR. The findings indicate that N. ceranae appears to be the dominant species in the Azores. N. apis was only detected in 2014/2015 with a very low prevalence (5.1%). N. ceranae prevalence varied between 2.7%, on São Jorge, and 50.7%, on Pico. In 2020, N. ceranae positive colonies increased significantly on Terceira (57.1%) and São Jorge (50.0%). N. ceranae was not detected on Santa Maria in both periods. Average infection levels in positive N. ceranae colonies were usually medium to high (>10-7 ng/μl), with São Jorge colonies displaying the greatest infection intensity (>10-5 ng/ μl). This study highlights the Azores archipelago as a unique place for beekeeping, with islands free of N. ceranae and V. destructor, which are two important stressors that afflict honey bees in the world.Financial support was provided by Portuguese funds through FCT (Fundação para a Ciência e a Tecnologia) in the framework of the project BeeHappy (POCI-01-0145-FEDER-029871). ARL was supported by a PhD scholarship (SFRH/BD/143627/2019) from the FCT. FCT provided financial support by national funds (FCT/MCTES) to CIMO (UIDB/00690/2020).info:eu-repo/semantics/publishedVersio