Antimicrobial resistance patterns of Staphylococcus spp. isolated from clinical specimens of companion animals in Northern Portugal, 2021-2023.

Staphylococcus spp. are growing pathogens in humans and companion animals. The emergence of multidrug-resistant bacterial infections, such as methicillin-resistant Staphylococcus-associated infections, due to zoonotic transmission, is a major public health concern. Domestic animals, such as dogs and cats, are possible reservoirs of multi-resistant bacterial species, which makes it relevant to monitor them due to their proximity to humans. However, there is a lack of information on the real scenario in Europe, especially in Portugal, particularly for animal infections caused by Staphylococcus spp. Therefore, this study aimed to investigate the antimicrobial resistance profile of Staphylococcus spp. isolated from cats and dogs diagnosed with infection in Northern Portugal. During 2021-2023, 96 Staphylococcus isolates from dogs and cats with symptoms of bacterial infection, including animals being treated in veterinary clinics/hospitals and cadavers submitted for necropsy at INIAV were included in the study collection. Of the 96 isolates, 63 were from dogs and 33 were Staphylococcus spp. from cats, most of which were isolated from ear (57% and 18%, respectively), skin (19 % and 27 %, respectively) and respiratory tract infections (6 % and 27 %, respectively). Among all the isolates, 12 different Staphylococcus spp. were identified, with Staphylococcus pseudintermedius being the most identified (61 % from dogs and 30 % from cats). It is noteworthy that 36 % of the isolates were multi-drug resistant and 25 % of the isolates showed a methicillin-resistant phenotype, with the mecA gene having been identified in all these isolates. This study highlights a high occurrence of multidrug-resistant Staphylococcus spp. in companion animals in Northern Portugal. This underlines the potential for cats and dogs to act as reservoirs of antimicrobial resistance, that can be transmitted to humans, posing a serious threat to public health.

An insight into seed priming response of Crotalaria ochroleuca and Crotalaria spectabilis during storage.

The proper establishment of plants is essential for the efficient use of resources such as water and light. Besides, even after seed storage and sowing the uniform establishment of plants is essential for their success. Crotalaria ochroleuca and Crotalaria spectabilis are important medicinal plants with poor seed germination rate, occasionally. The effects of seed priming in both C. ochroleuca and C. spectabilis were evaluated in seed performance even after seeds storage for up 90-days. Experimental assays were performed in a randomized design with gibberellic acid (GA3, 100 ppm), polyethylene glycol (PEG 6000, -0.2 MPa) and PEG (-0.2 MPa) + GA3 (100 ppm) solutions during seed priming in four replicates. Seeds not submitted to priming treatments constituted control. Seeds physiological performance were evaluated immediately and even after 30, 60 and 90-days seed dry-storage. The data obtained in each experiment were submitted to variance analysis (ANOVA) adopting a confidence level of 95%. The effects of seed priming with PEG and GA3 during seed ageing were significant for germination variables of C. ochroleuca and C. spectabilis. During dry storage, seed viability of both species gradually decreased and the first symptoms were delayed seed germination, especially more evident for C. ochroleuca, even in primed or non-primed seeds. Afterwards, C. ochroleuca seeds previously GA3 primed had higher results of root protrusion (86%), hypocotyls elongation (76%) and complete seedlings (75%) than non-primed seeds (control). These findings shown a good potential of hormopriming to attenuate damage during the seed aging of C. ochroleuca.

Comorbilidades en la esclerosis múltiple y su influencia en la elección del tratamiento / Comorbidities in multiple sclerosis and their influence on the choice of treatment

Los estudios clínicos de tratamientos para personas con esclerosis múltiple (pEM) se realizan en poblaciones seleccionadas, que excluyen a pacientes que presenten comorbilidades o medicaciones concomitantes. Sin embargo, un gran porcentaje de las pEM tiene alguna enfermedad adicional, que podría afectar a la respuesta y la elección del tratamiento. El objetivo de esta revisión es valorar cómo pueden las diferentes patologías concurrentes impactar en la elección de las terapias modificadoras de la enfermedad (TME) en las pEM. Se seleccionaron artículos relevantes mediante búsqueda en PubMed. Las comorbilidades se agruparon, a los fines de mejor ordenamiento de los artículos encontrados, en patologías diversas: autoinmunes, infecciones crónicas, cardiovasculares, respiratorias, metabólicas, oncológicas, neuropsiquiátricas y epilepsia. En cuanto a las patologías autoinmunes, es clave tener en cuenta los efectos de las TME sobre ellas y la posibilidad de interacción con sus tratamientos específicos. Las terapias inmunomoduladoras son seguras para personas con infecciones crónicas. Los tratamientos inmunosupresores, en general, están contraindicados en personas con infecciones activas. En las comorbilidades cardiovasculares y metabólicas deben tenerse en cuenta las potenciales reacciones de infusión asociadas a anticuerpos monoclonales, y los fenómenos asociados al inicio de tratamiento con moduladores del receptor de la esfingosina-1-fosfato. Las TME con efecto inmunosupresor están contraindicadas en personas con malignidades activas. Aunque la patología psiquiátrica de por sí no impide el uso de TME, debería tenerse precaución cuando aparecen nuevos síntomas psiquiátricos, y siempre tenerse en cuenta su monitorización y tratamiento. Por este motivo, entre los múltiples factores que deben considerarse a la hora de iniciar o cambiar una TME en pEM, las comorbilidades constituyen un elemento muchas veces decisivo.(AU)

Clinical trials of disease-modifying therapies (DMTs) for people with multiple sclerosis (pMS) are conducted in selected populations, excluding patients with comorbidities or concomitant medications. However, a large percentage of pMS have some additional disease, which could affect the response and choice of the DMT. The objective of this review is to assess how concurrent pathologies can impact the choice of DMTs. Relevant articles were selected through a systematic search in PubMed. Comorbidities were grouped for better classification into autoimmune, chronic infections, cardiovascular and metabolic, oncological and neuropsychiatric. In autoimmune pathologies, it is key to take into account the effects of TME on them and the possibility of interaction with their specific treatments. Immunomodulatory therapies are safe for people with chronic infections. Immunosuppressive treatments are generally contraindicated in people with active infections. In cardiovascular and metabolic comorbidities, infusion reactions associated with monoclonal antibodies, and the phenomena of starting treatment with S1P modulators, must be taken into account. DMTs with an immunosuppressive effect are contraindicated in people with active malignancies. Although psychiatric pathology per se does not preclude the use of DMTs, caution should be exercised when new psychiatric symptoms appear. For these reasons, among the multiple factors that must be considered when starting or changing a DMT in pMS, comorbidities constitute a decisive element.(AU)

[Comorbidities in multiple sclerosis and their influence on the choice of treatment]. / Comorbilidades en la esclerosis múltiple y su influencia en la elección del tratamiento.

Clinical trials of disease-modifying therapies (DMTs) for people with multiple sclerosis (pMS) are conducted in selected populations, excluding patients with comorbidities or concomitant medications. However, a large percentage of pMS have some additional disease, which could affect the response and choice of the DMT. The objective of this review is to assess how concurrent pathologies can impact the choice of DMTs. Relevant articles were selected through a systematic search in PubMed. Comorbidities were grouped for better classification into autoimmune, chronic infections, cardiovascular and metabolic, oncological and neuropsychiatric. In autoimmune pathologies, it is key to take into account the effects of TME on them and the possibility of interaction with their specific treatments. Immunomodulatory therapies are safe for people with chronic infections. Immunosuppressive treatments are generally contraindicated in people with active infections. In cardiovascular and metabolic comorbidities, infusion reactions associated with monoclonal antibodies, and the phenomena of starting treatment with S1P modulators, must be taken into account. DMTs with an immunosuppressive effect are contraindicated in people with active malignancies. Although psychiatric pathology per se does not preclude the use of DMTs, caution should be exercised when new psychiatric symptoms appear. For these reasons, among the multiple factors that must be considered when starting or changing a DMT in pMS, comorbidities constitute a decisive element.

TITLE: Comorbilidades en la esclerosis múltiple y su influencia en la elección del tratamiento.Los estudios clínicos de tratamientos para personas con esclerosis múltiple (pEM) se realizan en poblaciones seleccionadas, que excluyen a pacientes que presenten comorbilidades o medicaciones concomitantes. Sin embargo, un gran porcentaje de las pEM tiene alguna enfermedad adicional, que podría afectar a la respuesta y la elección del tratamiento. El objetivo de esta revisión es valorar cómo pueden las diferentes patologías concurrentes impactar en la elección de las terapias modificadoras de la enfermedad (TME) en las pEM. Se seleccionaron artículos relevantes mediante búsqueda en PubMed. Las comorbilidades se agruparon, a los fines de mejor ordenamiento de los artículos encontrados, en patologías diversas: autoinmunes, infecciones crónicas, cardiovasculares, respiratorias, metabólicas, oncológicas, neuropsiquiátricas y epilepsia. En cuanto a las patologías autoinmunes, es clave tener en cuenta los efectos de las TME sobre ellas y la posibilidad de interacción con sus tratamientos específicos. Las terapias inmunomoduladoras son seguras para personas con infecciones crónicas. Los tratamientos inmunosupresores, en general, están contraindicados en personas con infecciones activas. En las comorbilidades cardiovasculares y metabólicas deben tenerse en cuenta las potenciales reacciones de infusión asociadas a anticuerpos monoclonales, y los fenómenos asociados al inicio de tratamiento con moduladores del receptor de la esfingosina-1-fosfato. Las TME con efecto inmunosupresor están contraindicadas en personas con malignidades activas. Aunque la patología psiquiátrica de por sí no impide el uso de TME, debería tenerse precaución cuando aparecen nuevos síntomas psiquiátricos, y siempre tenerse en cuenta su monitorización y tratamiento. Por este motivo, entre los múltiples factores que deben considerarse a la hora de iniciar o cambiar una TME en pEM, las comorbilidades constituyen un elemento muchas veces decisivo.

Evaluation of the antimicrobial activity of Eucalyptus radiata essential oil against Escherichia coli strains isolated from meat products.

The present study sought to evaluate the antimicrobial and anti-adherent potential of Eucalyptus radiata essential oil against food-borne strains of Escherichia coli. The study was performed using the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC). In addition, the disk diffusion technique was used to evaluate the association of Eucalyptus radiata essential oil with synthetic antimicrobials. The Minimum Inhibitory Adherence Concentration (MIC) was also performed. The results revealed that E. radiata showed antimicrobial activity against the E. coli strains tested, with MIC values ranging from 500 µg/mL to 1000 µg/mL and MBC values ranging from 500 µg/mL to 1,024 µg/mL. As for the associations, it was observed that E. radiata oil exhibited a synergistic effect for some antibiotics, especially Ceftriaxone, with greater interference from the essential oil. Furthermore, it was effective in inhibiting the adherence of bacterial strains of E. coli, showing a more significant antibiofilm effect than the antibacterial agent 0.12% chlorhexidine digluconate. In summary, the essential oil of E. radiata showed antimicrobial potential against strains of E. coli of food origin, and can therefore, through in-depth studies, be used alone or in association with synthetic antimicrobials to combat infections caused by this pathogen.

Use of Household Insecticides against Mosquitoes in Dengue-Endemic Areas in Sri Lanka.

The use of household insecticides in the formulation of mosquito coils and vaporizers is common among the public to protect against mosquito-borne diseases. However, information regarding the use of these commonly practiced household insecticides is scarce in Sri Lanka. A questionnaire-based survey was directed to 600 households in Colombo, Gampaha, Kandy, Kalutara, Jaffna, and Galle districts in Sri Lanka. Household insecticides were used by the majority of the respondents (57.0%), and electric liquid vaporizers (46.7%) and mosquito coils (45.7%) were the commonly used forms. The active ingredients in mosquito coils were d-trans-allethrin (w/w 0.12%), metofluthrin (w/w 0.005%), and dimefluthrin (w/w 0.01%); in liquid vaporizers, they were prallethrin (w/v 1.24% and w/v 1.26%) and transfluthrin (w/v 0.9%). The majority of the household insecticide consumers were moderately satisfied with the effectiveness of the products (68.6%), and most of the participants used household insecticides daily (35.5%), especially during the evening (41.6%). Most of the consumers were unaware of the chemicals included in household insecticides (62.8%). Mosquito coils were abundantly used by rural area participants (75.8%), whereas liquid vaporizers were the most common among the urban participants (56.4%). The findings indicate that demographic and socioeconomic factors influence household insecticide practices. To our knowledge, this study reveals the use of household insecticides to control mosquitoes for the first time in Sri Lanka and highlights the importance of awareness programs and the proper utilization of these products.

The impact of sputum quality on Xpert positivity in active case-finding for TB.

BACKGROUND: Studies evaluating sputum quality and Xpert® MTB/RIF positivity in the context of active case finding are scarce. We aimed to determine whether sputum quality is associated with Xpert positivity and whether the association differed according to demographic and clinical characteristics.METHODS: A cross-sectional analysis using data from a mass screening programme in Brazilian prisons was conducted from 2017 to 2021. We administered a standardised questionnaire, obtained a chest X-ray and collected a spot sputum sample for Xpert testing. Sputum quality was classified as 'salivary', 'mucoid/mucopurulent' or 'blood-stained'. We used log binomial regressions to estimate the relationship between sputum quality and Xpert positivity, assessing interactions with participant characteristics.RESULTS: Among 4,368 participants for whom sputum quality was assessed, 957 (21.9%) produced salivary specimens, 3,379 (77.4%) had mucoid/mucopurulent sputum and 32 (0.7%) had blood-stained sputum. Xpert positivity was higher among those with mucoid/mucopurulent sputum than among those with salivary samples (12.0% vs. 3.7%). Mucopurulent sputum independently predicted Xpert positivity among individuals with and without symptoms, current smoking and abnormal chest radiographs on CAD4TB.CONCLUSIONS: In our study, sputum appearance independently predicted Xpert positivity, and could be used together with chest X-ray and symptom screening to inform use of Xpert in individual or pooled testing.

Fine-scale monitoring of insecticide resistance in Aedes aegypti (Diptera: Culicidae) from Sri Lanka and modeling the phenotypic resistance using rational approximation.

BACKGROUND: The unplanned and intensified use of insecticides to control mosquito-borne diseases has led to an upsurge of resistance to commonly used insecticides. Aedes aegypti, the main vector of dengue, chikungunya, and Zika virus, is primarily controlled through the application of adulticides (pyrethroid insecticides) and larvicides (temephos). Fine spatial-scale analysis of resistance may reveal important resistance-related patterns, and the application of mathematical models to determine the phenotypic resistance status lessens the cost and usage of resources, thus resulting in an enhanced and successful control program. METHODS: The phenotypic resistance for permethrin, deltamethrin, and malathion was monitored in the Ae. aegypti populations using the World Health Organization (WHO) adult bioassay method. Mosquitoes' resistance to permethrin and deltamethrin was evaluated for the commonly occurring base substitutions in the voltage-gated sodium channel (vgsc) gene. Rational functions were used to determine the relationship between the kdr alleles and the phenotypic resistant percentage of Ae. aegypti in Sri Lanka. RESULTS: The results of the bioassays revealed highly resistant Ae. aegypti populations for the two pyrethroid insecticides (permethrin and deltamethrin) tested. All populations were susceptible to 5% malathion insecticide. The study also revealed high frequencies of C1534 and G1016 in all the populations studied. The highest haplotype frequency was detected for the haplotype CC/VV, followed by FC/VV and CC/VG. Of the seven models obtained, this study suggests the prediction models using rational approximation considering the C allele frequencies and the total of C, G, and P allele frequencies and phenotypic resistance as the best fits for the area concerned. CONCLUSIONS: This is the first study to our knowledge to provide a model to predict phenotypic resistance using rational functions considering kdr alleles. The flexible nature of the rational functions has revealed the most suitable association among them. Thus, a general evaluation of kdr alleles prior to insecticide applications would unveil the phenotypic resistance percentage of the wild mosquito population. A site-specific strategy is recommended for monitoring resistance with a mathematical approach and management of insecticide applications for the vector population.

Relationship between sensory processing patterns and gross motor function of children and adolescents with Down syndrome and typical development: a cross-sectional study.

BACKGROUND: Children and adolescents with Down syndrome (DS) may experience impairments in sensory and motor skills that can be interrelated. The purposes of this study were (i) to compare the sensory processing patterns and gross motor function between children and adolescents with DS and with typical development (TD) and (ii) to explore associations between these areas in both DS and TD groups. METHOD: This cross-sectional study involved a sample size of 25 participants with DS (mean age 10.24 ± 2.04 years) and 25 participants with TD (mean age 10.04 ± 2.82 years). The sensory processing patterns were assessed using the Sensory Profile Second Version questionnaire, and the gross motor function was measured with the Gross Motor Function Measure (GMFM-88) dimensions (D) standing, and (E) walking, running, and jumping. Differences between groups were tested using the Mann-Whitney test, and the relationship between the variables was examined using Spearman's correlation tests, with a significance level set at 5%. RESULTS: Children with DS showed greater difficulties with sensory processing than TD children in Seeker (P < 0.001), Avoider (P < 0.001), Sensitivity (P < 0.001), Registration (P < 0.001), Auditory (P < 0.001), Touch (P = 0.001), Movements (P = 0.001), Oral (P = 0.028), Conduct (P = 0.005), Socioemotional (P < 0.001), and Attentional (P < 0.001) domains. Additionally, children with DS presented lower gross motor function than TD in GMFM-88, standing (P < 0.001) and walking, running, and jumping (P < 0.001). Correlations were found between greater difficulties with sensory processing in Touch and lower gross motor function in walking, running, and jumping for the DS group. CONCLUSION: Our results suggest there are more difficulties in sensory processing patterns and gross motor function in children with DS than in TD. Also, there is a single association between more difficulties in sensory processing and less well-developed motor function in the DS group. Therefore, a comprehensive assessment of all these aspects should be performed in children and adolescents with DS, along with the provision of relevant interventions addressing specific needs.

Sustainable permeable biobarriers for atrazine removal in packed bed biofilm reactors.

The synergy between two supported bacterial biofilms of S. equisimilis and P. putida and a sustainable biocarrier (raw pine) was studied, working both as biobarriers for the treatment of water contaminated with atrazine. Firstly, the effects of ATZ exposure on bacterial growth were evaluated, with Gram-positive S. equisimilis being a more tolerant bacterium to higher amounts of the herbicide. The bioremoval of ATZ by S. equisimilis concentrated biomass was then assessed, reaching around 83.5% after 15 days due to the potential degradation by the biomass and biosorption by the solids, with overlapping of both mechanisms. The optimization of bacterial biofilm attachment onto raw pine prior to bioremoval assays in lab-scale packed bed biofilm reactors was performed by varying initial biomass concentration, inocula growth time and hydrodynamic conditions. Lastly, the optimized biosystems were tested as sustainable remediation designs to treat water contaminated with the selected herbicide. Results reveal an added beneficial effect towards the bioremoval of atrazine using supported biofilms onto raw pine, reaching 90.42% and 79.71% by S. equisimilis and P. putida biofilms, respectively, over 58.31% increase when compared to sorption on fixed bed of pine. The coupling of biosorption/biodegradation favors the bioremoval process significantly.

Antimicrobial potential of extracts from leaves and culms of an Amazonian native bamboo.

Antibiotics have shown less efficiency against resistance of pathogenic microorganisms. As a result, research centers have sought therapeutic alternatives against multidrug resistance of bacteria to antibiotics, one of which is using plant extracts. Bamboo extracts are used for several medicinal purposes. This study aimed to evaluate the antibacterial potential of hydroalcoholic extracts of culms and leaves of the species Guadua aff. lynnclarkiae on strains of Staphylococcus aureus, Streptococcus pneumoniae, and Klebsiella pneumoniae. We evaluated the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC). Only the leaves of G. aff. lynnclarkiae showed a bactericidal effect for all tested strains with MBC ranging from 1.55 mg ml-1 to 25 mg ml-1. The culms had bacteriostatic action with MIC ranging from 1.55 mg ml-1 to 6.25 mg ml-1, and bactericidal action at the concentration of 6.25 mg ml-1 only for S. aureus. This study provides bases for the use of this Amazonian native bamboo in bioprospecting.

Systematic review of motivational interventions to improve adherence to medication in patients with hypertension and meta-analysis / Revisión sistemática de intervenciones motivacionales para mejorar la adherencia a la medicación en pacientes con hipertensión y metanálisis

Introduction: Antihypertensive medication non-adherence is an important cause of poor control in hypertension. The role of motivational interventions to increase antihypertensive medication adherence remains unclear. Objective: To systematically review RCTs of motivational interventions for improving medication adherence in hypertension. Methods: EMBASE and Pubmed were searched from inception to February 2019 for RCTs of motivational interventions for improving medication adherence in hypertension vs. usual care. Inclusion criteria: RCTs with motivational intervention to improve medication adherence in adults with hypertension. A blinded review was conducted by 2 reviewers. Disagreements were resolved by consensus/a third reviewer. Data extraction and quality appraisal was performed using the risk of bias tool from cochrane collaboration. The meta-analyses of blood pressure control used random-effects models to report mean difference and 95% CIs. Primary outcome was medication adherence and second outcome was blood pressure control. Results: The search methodology yielded 10 studies comprising 1171 participants. Medication adherence improved significantly in 5 studies. We could not perform pool analysis for this outcome due to different measurements of medication adherence. Seven trials reported significant results regarding blood pressure control. On pooled analysis, motivational interventions were not significantly associated with a systolic blood pressure (mean difference, −0.06; 95% CI, −0.05 to 0.18; p=0.63; I2=0.0%) or diastolic blood pressure (mean difference, −0.11; 95% CI, −0.10 to 0.31; p=0.28; I2=23.8%) decrease or blood pressure control. (AU)

Introducción: La falta de adherencia a la terapia farmacológica es una de las principales razones del descontrol de la hipertensión arterial. Se desconoce el papel de las intervenciones motivacionales en el aumento de la adherencia. Objetivo: Realizar una revisión sistemática de ensayos clínicos aleatorizados (ECA) dirigidos a mejorar la adherencia a la medicación en hipertensión arterial. Métodos: Se buscaron ECA de intervenciones motivacionales vs. atención habitual en las bases de datos Embase y PubMed desde su inicio hasta febrero de 2019. Criterios de inclusión: ECA de intervenciones motivacionales para aumentar la adherencia a la terapia con medicamentos en adultos con hipertensión. Dos revisores realizaron una revisión ciega y sus desacuerdos se resolvieron por consenso/por un tercer revisor. La extracción de datos y la evaluación de la calidad se realizaron mediante la herramienta Cochrane de evaluación del riesgo de sesgo. El metaanálisis del control de la presión arterial utilizó modelos de efectos aleatorios para informar la diferencia en las medias y los intervalos de confianza de 95% (IC 95%). El outcome primario fue la adherencia a la medicación y el secundario fue el control de la presión arterial. Resultados: Se obtuvieron 10 estudios con 1.171 participantes. La adherencia mejoró significativamente en cinco estudios. No fue posible realizar un análisis agrupado de la adherencia debido al uso de diferentes medidas de cumplimiento. Siete estudios mostraron una diferencia significativa en el control de la presión arterial. En el análisis conjunto, las intervenciones motivacionales no se asociaron a una disminución significativa de la presión arterial sistólica (diferencia de medias, -0,06; IC 95%, -0,05-0,18; p=0,63; I2=0%) o de la presión arterial diastólica (diferencia de medias, -0,11; IC 95%, -0,10-0,31; p=0,28; I2=23,8%) o a mejora en control de la misma.(AU)

Agronomic parameters of sugarcane under planting densities in different cultivation cycles.

Low density sugarcane plantation (LDSP) has been implemented by some sugarcane producers in Brazil, aiming to save seeds and operational costs. The study was carried out in the municipality of Areia, Paraíba, Brazil. Five planting densities were used, varying from 5 to 25 m-2 of buds arranged in randomized blocks, with four replications. Data were measured annually over three cultivation cycles (2017 to 2020), during which the field was fertilized with NPK and the harvests were carried out manually without prior burning. The lower planting density presents higher productivity only in the cane plant (101.03 t ha-1) due to the higher plant height (2.37 m) and the higher number of stalks (11 stalks m-2), suggesting that these variables are due to the greater availability of light, water and photosynthate. However, there is a drastic reduction in sugarcane yield for this lower population in the 2nd ratoon by up to 65.62%, which is correlated with number of stalks per meter. We demonstrate the agronomic viability of LDSP in the population of 10 buds m-2 in relation to conventional planting of sugarcane until the 2nd ratoon. Data are important for future studies to present additional considerations for other production factors, such as the effects of mechanized harvesting and the management of nutrients and water, assessing the sustainability of this large-scale planting system.

Genetic diversity and phylogeography of Phlebotomus argentipes (Diptera: Psychodidae, Phlebotominae), using COI and ND4 mitochondrial gene sequences.

BACKGROUND: Phlebotomus argentipes complex is the primary vector for cutaneous leishmaniasis, a burgeoning health concern in contemporary Sri Lanka, where effective vector control is important for proper disease management. Understanding the genetic diversity of the P. argentipes population in Sri Lanka is vital before implementing a successful vector control program. Various studies have indicated that genetic divergence, caused by genetic drift or selection, can significantly influence the vector capacity of arthropod species. To devise innovative control strategies for P. argentipes, exploring genetic diversity and phylogeography can offer valuable insights into vector competence, key genetic trait transfer, and impact on disease epidemiology. The primary objective is to analyze the genetic diversity and phylogeography of the P. argentipes complex in Sri Lanka, based on two mitochondrial genomic regions in modern representatives of P. argentipes populations. METHODOLOGY: A total of 159 P. argentipes specimens were collected from five endemic areas of cutaneous leishmaniasis and identified morphologically. Two mitochondrial regions (Cytochrome c oxidase subunit I (COI) and NADH dehydrogenase subunit 4 (ND4) were amplified using the total DNA and subsequently sequenced. Partial sequences of those mitochondrial genes were utilized to analyze genetic diversity indices and to explore phylogenetic and phylogeographic relationships. PRINCIPAL FINDINGS: Among five sampling locations, the highest genetic diversity for COI and ND4 was observed in Hambantota (Hd-0.749, π-0.00417) and Medirigiriya (Hd-0.977, π-0.01055), respectively. Phylogeographic analyses conducted using COI sequences and GenBank retrieved sequences demonstrated a significant divergence of P. argentipes haplotypes found in Sri Lanka. Results revealed that they have evolved from the Indian ancestral haplotype due to historical- geographical connections of the Indian subcontinent with Sri Lanka. CONCLUSIONS: Utilizing high-mutation-rate mitochondrial genes, such as ND4, can enhance the accuracy of genetic variability analysis in P. argentipes populations in Sri Lanka. The phylogeographical analysis of COI gene markers in this study provides insights into the historical geographical relationship between India and P. argentipes in Sri Lanka. Both COI and ND4 genes exhibited consistent genetic homogeneity in P. argentipes in Sri Lanka, suggesting minimal impact on gene flow. This homogeneity also implies the potential for horizontal gene transfer across populations, facilitating the transmission of genes associated with traits like insecticide resistance. This dynamic undermines disease control efforts reliant on vector control strategies.

Trapping Microparticles in a Structured Dark Focus.

We experimentally demonstrate stable trapping and controlled manipulation of silica microspheres in a structured optical beam consisting of a dark focus surrounded by light in all directions-the dark focus tweezer. Results from power spectrum and potential analysis demonstrate the nonharmonicity of the trapping potential landscape, which is reconstructed from experimental data in agreement to Lorentz-Mie numerical simulations. Applications of the dark tweezer in levitated optomechanics and biophysics are discussed.

Regulation of dormancy break and germination of safflower seeds: the role of GA3, light and cold temperatures.

The safflower crop is considered a great alternative for crop rotation since drought tolerance and low production cost are attractive for its choice. However, safflower seeds show dormancy soon after dispersal from the mother plant, making it difficult to successfully establish plants using newly harvested seeds. The influence of temperature, gibberellin and light/dark on dormancy break of safflower seeds during storage were investigated. In a completely randomized design, freshly harvested seeds or stored for 100 and 200 days (paper bag, 20 °C/ 60% UR) were treated with GA3 (0 and 100 µM), at 4, 10 and 25 °C, in the presence and absence of light, during the germination test. Seeds were evaluated for germination percentage, germination speed and seedling survival after 21 days. The temperature of 10 °C, in combination with GA3 (0/100 µM), or light/dark, provided the highest seed germination results, for freshly-harvested seeds and stored seeds. Collectively, these observations indicate that dormancy was not affected by gibberellic acid (100 µM GA3) and the germination results at 21 days were significantly higher, in relation to the use of GA3, under light or dark. Recently harvested seeds could efficiently germinate at 10 °C in the dark, while seeds dry-stored at 20 °C had decreased germination percentages.

Comparison of non-aureus staphylococcal and mammaliicoccal species found in both composite milk and bulk-tank milk samples of dairy cows collected in tandem.

Non-aureus staphylococci and the closely related mammaliicoccal species (NASM) are the most common causes of bovine subclinical mastitis on modern dairy farms and are highly prevalent in bulk-tank milk. The purpose of this study was to determine the distribution of NASM in both composite cow milk (CCM) and bulk-tank milk (BTM) samples collected in tandem in commercial Flemish dairy herds and to estimate the origin of the different (subgroups of) NASM species present in BTM by applying strain typing (random amplification of polymorphic DNA or random amplified DNA [RAPD]). A single cross-sectional sampling was performed over 5 herds that volunteered to participate in the study. Composite cow milk samples (n = 356) were collected from all lactating cows (except those with clinical mastitis) during a milking in tandem with 6 BTM samples per herd sequentially collected immediately post that milking (n = 30). In total, 421 and 80 NASM isolates were recovered and identified by MALDI-TOF mass spectrometry from the CCM and BTM samples, respectively and a total of 21 and 12 different NASM species were identified from CCM and BTM samples, respectively. Staphylococcus cohnii was the most prevalent NASM species found in BTM followed by Staphylococcus haemolyticus, Staphylococcus epidermidis, Mammaliicoccus lentus, and Staphylococcus equorum, whereas from CCM samples the most common species were S. hemolyticus, S. cohnii, S. equorum, S. epidermidis, and Staphylococcus chromogenes. The prevalent NASM species in both CCM and BTM samples was distinct for each herd, corroborating other studies observing a herd-specific NASM microbiota. Random amplified DNA analysis was performed on 9 NASM species (S. chromogenes, S. epidermidis, S. haemolyticus, S. equorum, Mammaliicoccus sciuri, Staphylococcus xylosus, S. cohnii, Staphylococcus debuckii, and M. lentus) because these species were isolated from both sample types in a herd. The same RAPD types were found in both sample types for all NASM species selected for strain typing in varying degrees. When assessing the distribution of NASM species, differences within NASM species should be examined meaning a closer look should be taken at the strain level rather than at the species level only.

Age-related changes in circulating INSL3 concentrations and their associations with ovarian conditions in Japanese Black beef cattle.

Information on circulating levels of insulin-like peptide 3 (INSL3) in female domesticated animals is limited, as their concentrations are significantly lower than in males. The objectives of the present study were to 1) develop a sandwich time-resolved fluorescence immunoassay (TRFIA) with higher detectability to measure blood INSL3 concentrations in female cattle, 2) determine INSL3 concentrations in female cattle among age groups and reproductive conditions, and 3) explore associations between INSL3 levels and ultrasonographic ovarian measurements. Blood was collected repeatedly from Japanese Black beef female calves (n = 12; 0－8 mo), heifers (n = 10; 10－26 mo), and cows (n = 20; 27－200 mo). Blood was taken from the cows (n = 13) at follicular, post-ovulatory, and luteal phases, and from cows with follicular cysts (n = 12). Ultrasonography of ovaries was conducted in the calves (n = 12) and the cows without ovarian diseases (n = 9). The ovarian area, as well as the number and diameters of antral follicles ≥ 2 mm, were determined in each ovary. The proposed method detected a difference in plasma INSL3 between calves (0.01 ng/mL) and heifers (0.18 ng/mL). However, the conventional assay showed similar levels for calves and heifers (1.82 vs 2.07 ng/mL). Plasma INSL3 and testosterone concentrations increased from calves to heifers (P < 0.0001), but only INSL3 rose from heifers to cows (P < 0.0001). INSL3 and testosterone concentrations did not change across the estrus cycle in cows, and the levels of both hormones in follicular cystic cows did not differ from those in the follicular phase. Ovarian area, maximal and average follicular diameters, and total volume of all follicles per animal were higher in cows than calves (P < 0.001). Plasma INSL3 concentrations correlated positively with the total volumes of all follicles in calves (P < 0.05) and cows (P < 0.05), whereas testosterone concentrations did not correlate with ovarian follicular measurements. In conclusion, plasma INSL3 concentrations measured by the proposed sandwich TRFIA showed a clear increase from female calves to cows in beef cattle. These results suggest that circulating levels of INSL3, but not of testosterone, are associated with the total volume of all antral follicles in both ovaries per animal in female cattle.

Genetic diversity, phylogenetic and phylogeographic analysis of Anopheles culicifacies species complex using ITS2 and COI sequences.

Anopheles culicifacies is the major vector of malaria in Sri Lanka and the Indian subcontinent which is characterized as a species complex with five sibling species provisionally designated as A, B, C, D and E. The current study was carried out to understand the phylogenetic and phylogeographic relationships between the sibling species of the species complex while observing their genetic diversity and genetic differentiation. Thirty-five ITS2 and seventy-seven COI sequences of An. culicifacies species complex reported from different geographical locations of Asia and China at the NCBI public database were used for the analysis. Bayesian likelihood trees were generated for the phylogenetic analysis. The divergence of the species complex was obtained from the Bayesian phylogeographic model in BEAST. There were two clades of the sibling species of An. culicifacies species complex as A, D and B, C and E in both phylogenetic and phylogeographic analysis using ITS2 sequences. Based on the highly divergent COI sequences and the high mutation rate of the mitochondrial genome, there were four and three clades in both phylogenetic and phylogeographic analysis using COI sequences. The diversification of An. culicifacies species complex was obtained as ranging from 20.25 to 24.12 Mya and 22.37 to 26.22 Mya based on ITS2 and COI phylogeographic analysis respectively. There was a recent diversification of the sibling species A and D than the sibling species B, C and E. Low haplotype diversity was observed in the sequences reported from Sri Lanka in both ITS2 and COI analysis that can be due to bottlenecks resulting from the intense malaria control efforts. A high genetic differentiation was achieved for some populations due to the large geographical distance. The high genetic diversity based on the five sibling species implies the possibility of maintaining a relatively high effective population size despite the vector control efforts.

Helminth community structure of the white-bellied woolly mouse opossum Marmosa constantiae Thomas, 1904 in Central-West Brazil.

Marmosa constantiae is a species of marsupial restricted to the central portion of South America. In Brazil, it occurs in the northwestern region including five states of the Amazon, Cerrado, and Pantanal biomes. However, there is no study of the helminth fauna or helminth community structure for this marsupial. The aims of this study were to describe the species composition and to analyse the structure of the helminthic community of M. constantiae in an area of the Amazon Arc in Sinop, north of the state of Mato Grosso, Brazil. Parasites were searched in 53 specimens of this marsupial, among which 44 were infected with at least one helminth species. Parasitic helminths were counted and identified. Nine species were collected: seven nematodes, one cestode, and one acanthocephalan. The most abundant species were Gracilioxyuris agilisis, Travassostrongylus scheibelorum, Pterygodermatites sinopiensis, and Subulura eliseae. These species were the only dominant ones in the component community. No significant differences were observed in the abundance and prevalence of helminths between male and female hosts. Host body size significantly influenced helminth abundance in males. The pattern of community structure considering the infracommunities in this locality indicated more species replacement than species loss along the environmental gradient. This is the first study to report the helminth fauna and the helminth community structure of M. constantiae.