Molecular Epidemiology of Plasmodium species in Conflicted Federally Administered Tribal Area (FATA) Pakistan

Military conflicts have been significant obstacles in detecting and treating infectious disease diseases due to the diminished public health infrastructure, resulting in malaria endemicity. A variety of violent and destructive incidents were experienced by FATA (Federally Administered Tribal Areas). It was a struggle to pursue an epidemiological analysis due to continuing conflict and Talibanization. Clinical isolates were collected from Bajaur, Mohmand, Khyber, Orakzai agencies from May 2017 to May 2018. For Giemsa staining, full blood EDTA blood samples have been collected from symptomatic participants. Malaria-positive microscopy isolates were spotted on filter papers for future Plasmodial molecular detection by nested polymerase chain reaction (nPCR) of small subunit ribosomal ribonucleic acid (ssrRNA) genes specific primers. Since reconfirming the nPCR, a malariometric study of 762 patients found 679 positive malaria cases. Plasmodium vivax was 523 (77%), Plasmodium falciparum 121 (18%), 35 (5%) were with mixed-species infection (P. vivax plus P. falciparum), and 83 were declared negative by PCR. Among the five agencies of FATA, Khyber agency has the highest malaria incidence (19%) with followed by P. vivax (19%) and P. falciparum (4.1%). In contrast, Kurram has about (14%), including (10.8%) P. vivax and (2.7%) P. falciparum cases, the lowest malaria epidemiology. Surprisingly, no significant differences in the distribution of mixed-species infection among all five agencies. P. falciparum and P. vivax were two prevalent FATA malaria species in Pakistan’s war-torn area. To overcome this rising incidence of malaria, this study recommends that initiating malaria awareness campaigns in school should be supported by public health agencies and malaria-related education locally, targeting children and parents alike.

Benefits and Costs of Mixed-Species Aggregations in Harvestmen (Arachnida: Opiliones)

Many animals form aggregations with individuals of the same species (single-species aggregations, SSA). Less frequently, individuals may also aggregate with individuals of other species (mixed-species aggregations, MSA). Although the benefits and costs of SSA have been intensively studied, the same is not true for MSA. Here, we first review the cases of MSA in harvestmen, an arachnid order in which the records of MSA are more frequent than other arthropod orders. We then propose several benefits and costs of MSA in harvestmen, and contrast them with those of SSA. Second, using field-gathered data we describe gregariousness in seven species of Prionostemma harvestmen from Costa Rica. These species form MSA, but individuals are also found solitarily or in SSA. We tested one possible benefit and one possible cost of gregariousness in Prionostemma harvestmen. Regarding the benefit, we hypothesized that individuals missing legs would be more exposed to predation than eight-legged individuals and thus they should be found preferentially in aggregations, where they would be more protected from predators. Our data, however, do not support this hypothesis. Regarding the cost, we hypothesized that gregariousness increases the chances of parasitism. We found no support for this hypothesis either because both mite prevalence and infestation intensity did not differ between solitary or aggregated individuals. Additionally, the type of aggregation (SSA or MSA) was not associated with the benefit or the cost we explored. This lack of effect may be explained by the fluid membership of the aggregations, as we found high turnover over time in the number of individuals and species composition of the aggregations. In conclusion, we hope our review and empirical data stimulate further studies on MSA, which remains one of the most elusive forms of group living in animals.

Optimized splitting of mixed-species RNA sequencing data

Gene expression studies using xenograft transplants or co-culture systems, usually with mixed human and mouse cells, have proven to be valuable to uncover cellular dynamics during development or in disease models. However, the mRNA sequence similarities among species presents a challenge for accurate transcript quantification. To identify optimal strategies for analyzing mixed-species RNA sequencing data, we evaluate both alignment-dependent and alignment-independent methods. Alignment of reads to a pooled reference index is effective, particularly if optimal alignments are used to classify sequencing reads by species, which are re-aligned with individual genomes, generating [Formula: see text] accuracy across a range of species ratios. Alignment-independent methods, such as convolutional neural networks, which extract the conserved patterns of sequences from two species, classify RNA sequencing reads with over 85% accuracy. Importantly, both methods perform well with different ratios of human and mouse reads. While non-alignment strategies successfully partitioned reads by species, a more traditional approach of mixed-genome alignment followed by optimized separation of reads proved to be the more successful with lower error rates.

Efficacy of bakuchiol-garlic combination against virulent genes of Candida albicans

Background Polymicrobial biofilms are notorious for causing intraoral tissue destruction. Streptococcus sanguinis and Streptococcus mitis, commensals of oral cavities, have been found co-existing with C. albicans in resistant oral infections. There is an urgent need to find alternative treatment options. This study aims to assess the efficacy of garlic (G) and bakuchiol (Bk) combination against candida virulent genes and their subsequently secreted proteins. Methods In vitro single species biofilms of C. albicans, and mixed species biofilms formed in combination with streptococci were exposed to bakuchiol and garlic extract (Bk+G). Gene expression of agglutinin-like sequence (ALS1), (ALS3), adhesin-like wall proteins (HWP1) and aspartyl proteinases (SAP5) were determined using qPCR and their subsequent proteins were assessed through Western blotting. Results Virulent genes were significantly downregulated in single species biofilms when they were treated with Bk+G combination. However, Bk+G did not have significant effect on ALS1 and HWP1 gene in polymicrobial biofilms. ALS3 and SAP5 were significantly downregulated in Bk+G treated polymicrobial biofilm. Similar results were portrayed in Western blotting. Conclusion Bk+G combination exhibited antimicrobial effects against single and mixed species biofilms. The findings might provide insights for treating resistant candida infections. This combination could potentially serve as an herbal alternative to traditional antifungals following further research.

Recovery of Candida spp. from a denture rat model

Denture stomatitis is a common infection in denture wearers. This study evaluated the recovery of Candida spp. from the palate of Wistar rats after using an acrylic device with single and mixed-species of Candida spp. After approval of the Ethics Committee, 84 male and female Wistar rats were used. Custom-made acrylic devices were fabricated for each animal and sterilized by microwave irradiation. Single and mixed species biofilms of C. albicans (Ca), C. glabrata (Cg), and C. tropicalis (Ct) were grown on the devices for 48 h at 37°C. Rats were anesthetized and the devices were cemented on the molar teeth (n=5 for each sex and Candida spp.). Rats received a carbohydrate-rich diet. Single and mixed species were inoculated in the oral cavity thrice after three-day intervals. Controls received only dentures without Candida spp. After 4 weeks, the devices were removed, the palates were swabbed, and diluted samples were plated on Agar Sabouraud Dextrose and CHROMAgar Candida for colony counting and presumptive identification, respectively, after 48 h. Data were analyzed by 3way ANOVA (α=5%). There was a significant interaction (p=0.003) between sex and species. For females, all groups recovered significant values (p≤0.027) compared with controls. For males, groups with Ct as single and dual-species showed the lowest values without difference (p≥0.183) with the control. The groups with triple-species showed the highest values but without difference (p≥0.071) with the groups with single and dual-species, except males with Ct. Ct alone showed reduced recovery from palate of male rats.

Does the Introduction of N2-Fixing Trees in Forest Plantations on Tropical Soils Ameliorate Low Fertility and Enhance Carbon Sequestration via Interactions Between Biota and Nutrient Availability? Case Studies From Central Africa and South America

Plant and/or crop growth rely on nutrient dynamics driven by specific soil biota in different environments. This mini-review aims to provide an overview of interactions between soil organisms, nutrient dynamics, and C sequestration. To this end, we investigated published results from three forest plantations (eucalyptus monocultures and mixed plantations with N2-fixing acacia) on tropical nutrient-poor soils. One case study is located in Central Africa (Congolese coastal plains) and two others in South America (Southeastern Brazil). Overall, the studies showed that soil biota activity exerted positive effects on (i) C accretion, as both soil carbon and belowground and aboveground biomass are driven and enhanced by soil biota; and (ii) on nutrient dynamics and biogeochemical cycles in nutrient-poor soil of tropical ecosystems, which are boosted following C accumulation. On the other hand, the pedoclimatic environment may potentially impact soil functioning of mixed-species plantations through its influence on the composition and activity of bacterial communities. Regardless of the potential risk of acacia invasiveness, benefits such as pulp, fuelwood, electric pole and non-timber products supply, have been reported in Central Africa. We, therefore, conclude that including N2 fixing trees in forestry plantations as reported in this mini-review helps strengthen the links between soil biota, nutrient and SOC dynamics in mixed-species plantations on tropical nutrient-poor soils.