Ethnobotanical relevance: Piper guineense occurs commonly in West Africa where it is used for fungal infections instead of the costly and not always accessible conventional antifungals. Fungal, yeast-based diseases are common in West-Africa especially among those living with HIV/AIDS, and thus this study was performed in Imo state, South-Eastern Nigeria, where P. guineense is predominantly used for the treatment of fungal diseases, such as skin rashes, oral thrush and vaginosis. Aim of study: The scarce number of previous studies on the documentation of the traditional uses of P. guineense extracts for the treatment of fungal infections in Nigeria prompted this survey. The investigation focused on how traditional healers recognize and diagnose fungal infections, how P. guineense is collected, on the various parts used for the treatments, methods of preparations, administrations and treatments. In addition, an in vitro anti fungal screening of P. guineense fruit and leaf extracts of various polarities, and piperine and piperlongumine, representing the main constituents in these extracts, were performed. Methods: A house to house ethnobotanical survey was conducted using questionnaires. Twenty traditional medical practitioners (TMP) and herb sellers from ten villages were interviewed. Four human pathogenic strains of yeast and Cryptococcus neoformans, a yeast-like basidiomycete causing meningitis in immunocompromised individuals, were used for the antifungal screening. Results: The traditional medical practitioners (TMP) and herb sellers explained that the leaves and fruits are the most commonly used plant parts for the treatments. The oral intake of the extracts in locally produced bamboo alcohol (Kai-kai) is the most common method of administration. In accordance with these recorded traditional uses, we found that extracts of P. guineense were growth inhibitory against the fungal strains with MIC values ranging from 39 to 2500 g/mL. The lowest MIC value of 39 g/mL was recorded for a methanol fruit extract against Candida albicans, C. glabrata and C. tropicalis. In addition, ethanol and hexane fruit extracts were effective against the growth of C. albicans and C. glabrata, respectively, with a MIC of 78 g/mL. Piperlongumine and piperine were active against C. albicans with MIC values of 39 and 78 g/mL respectively. Conclusion: P. guineense fruit and leaf extracts, as well as their piperamide alkaloid constituents piperine and piperlongumine, have interesting antifungal properties and could have potential as new antifungal scaffolds. Our results warrant further in-depth investigations to isolate and characterize piperamide alkaloids and other compounds responsible for the antifungal activity in the extracts.

Diarrheal disease, still a major cause of childhood illness, is caused by numerous, diverse infectious microorganisms, which are differentially sensitive to environmental conditions. Enteropathogen-specific impacts of climate remain underexplored. Results from 15 studies that diagnosed enteropathogens in 64,788 stool samples from 20,760 children in 19 countries were combined. Infection status for 10 common enteropathogens-adenovirus, astrovirus, norovirus, rotavirus, sapovirus, Campylobacter, ETEC, Shigella, Cryptosporidium and Giardia-was matched by date with hydrometeorological variables from a global Earth observation dataset-precipitation and runoff volume, humidity, soil moisture, solar radiation, air pressure, temperature, and wind speed. Models were fitted for each pathogen, accounting for lags, nonlinearity, confounders, and threshold effects. Different variables showed complex, non-linear associations with infection risk varying in magnitude and direction depending on pathogen species. Rotavirus infection decreased markedly following increasing 7-day average temperatures-a relative risk of 0.76 (95% confidence interval: 0.69-0.85) above 28 degrees C-while ETEC risk increased by almost half, 1.43 (1.36-1.50), in the 20-35 degrees C range. Risk for all pathogens was highest following soil moistures in the upper range. Humidity was associated with increases in bacterial infections and decreases in most viral infections. Several virus species' risk increased following lower-than-average rainfall, while rotavirus and ETEC increased with heavier runoff. Temperature, soil moisture, and humidity are particularly influential parameters across all enteropathogens, likely impacting pathogen survival outside the host. Precipitation and runoff have divergent associations with different enteric viruses. These effects may engender shifts in the relative burden of diarrhea-causing agents as the global climate changes. Plain Language Summary Diarrheal disease is a big health problem for children. It can be caused by different bugs, which can be caught more easily in certain weather conditions, though not much is understood about this because the climate varies so much from one place to the next. This study combined data from many different countries where diarrhea-causing bugs were diagnosed in children's stool. Satellites recorded what the weather was like on the day each sample was collected. Rotavirus is easiest to catch in cold weather and when water washes over the ground after rain. Dry weather also makes it and other viruses easy to catch. Bacteria spread best when the air is warm and humid, and the soil moist, though one type of E. coli can also be spread in rainwater. Climate change will make dry places drier, wet places wetter and everywhere warmer. This might lead to more diarrhea caused by bacteria and less by viruses in some places, though places with moist soil might see more of every kind of bug.

Background: Ticks of the group Rhipicephalus sanguineus (sensu lato) are distributed worldwide and are major pathogen vectors of both dogs and humans. Previous phylogenetic reconstructions have suggested the existence of two main lineages within this group, "Tropical" and "Temperate". Symbiotic interactions contribute to vector development, survival, reproduction and competence. The diversity of microbial communities associated with different populations of R. sanguineus (s.l.) remains poorly characterized, however, this knowledge will aid in future studies of hosts-microbiota-pathogen interactions. To gain insight into the bacterial communities associated with R. sanguineus (s.l.) ticks, 40 specimens from France, Senegal and Arizona were analyzed by high-throughput 16S amplicon sequencing. All tick specimens were taxonomically classified using the mitochondrial 12S rDNA gene, which provides sufficient phylogenetic resolution to discriminate different lineages of R. sanguineus. Results: Rhipicephalus sanguineus (s.l.) samples from Senegal belonged to the "Tropical" lineage, samples from France belonged to the "Temperate" lineage, whereas both lineages were identified in samples from Arizona. Regardless of origin, each bacterial microbiota was dominated by three genera: Coxiella, Rickettsia and Bacillus. Rickettsia and Coxiella were the two main genera found in females whereas males had a higher proportion of Bacillus. Significant differences of relative abundances were evidenced between specimens from different geographical origins. Conclusions: This study highlights differences in the microbiota composition within R. sanguineus (s.l.) specimens from different genotypes, genders and geographical origins. This knowledge will help in future studies of the symbiotic interactions, biology and vector competence of the R. sanguineus (s.l.) complex.

Background: Enterotoxigenic Escherichia coli (ETEC) is a major pathogen causing travellers' diarrhoea (TD) among visitors to low- and middle-income countries (LMIC). Scant data are available on rates of travel-acquired ETEC producing heat-labile (LT) and/or heat-stable (ST) toxin or its subtypes, STh (human) and STp (porcine) in various geographic regions, and on clinical pictures associated with each toxin. Methods: Using qPCR, we analysed LT, STh, and STp in stools positive for ETEC in a prospective study among 103 Finnish travellers visiting LMIC. They filled in questionnaires and provided stool samples before and after travel. We scrutinized geographic distribution of LT, STh, and STp ETEC findings, and association between these different ETEC subtypes and moderate/severe TD. Results: Among the 103 stool samples positive for ETEC toxins, the rate for LT was 76%, for STh 26%, and STp 41%. The rate for LT-only was 44%, for 5Th-only 6%, STp-only 16%, LT + STh 10%, LT + STp 15%, STh + STp 3%, and LT + STh + STp 8%. Findings varied by destination; the rates of LT, STh, and STp were 79%, 21%, and 57%, respectively, in Southern Asia (n = 14); 85%, 10%, and 20% in South-eastern Asia (n = 20); 84%, 13%, and 29% in Eastern Africa (n = 31); and 56%, 50%, and 63% in Western Africa (n = 32), respectively. Of travellers positive for LT, STh, and STp, 83%, 100%, and 88%, encountered TD; 35%, 55%, and 41% reported moderate/severe TD. STh was associated with moderate/severe TD. Conclusions: Toxin findings varied by destination; multiple toxins were commonly detected. Moderate/severe TD was reported most frequently by subjects with STh-ETEC.

Background and objectives: The global forest economy is threatened by eucalypt pathogens which are often latent or cryptic species that escape common quarantine and detection methods. Plantation forestry using eucalypts is of considerable importance to Paraguay, but knowledge regarding the pests and diseases affecting these plantations is limited. This study identified fungal diseases present in these plantations. Materials and Methods: We surveyed eucalypt plantations in four provinces in Paraguay and collected material from diseased trees for identification of the causal agents. The samples were analyzed using a combination of morphological and molecular methods. Results: Diseases encountered included Botryosphaeria stem canker, Calonectria leaf blight, Chrysoporthe stem canker, myrtle/eucalypt rust, Coniella leaf spot, heartwood rot and Teratosphaeria stem canker. Contrary to expectations, the causal agent of Teratosphaeria stem canker was identified as Teratosphaeria zuluensis (M.J. Wingf., Crous & T.A. Cout.) M.J. Wingf. & Crous and not Teratosphaeria gauchensis (M.-N. Cortinas, Crous & M.J. Wingf.) M.J. Wingf. & Crous, that is commonly documented for the South American region. Conclusions: This study updates the knowledge on forest fungal pathogens in Paraguayan eucalypt plantations and is the first report of T. zuluensis in Paraguay and in South America.

Most tick-related studies in Europe have been conducted in nonurban areas, but ticks and tick-borne pathogens also occur in urban green spaces. From a public health perspective, risks regarding tick-borne infections should be studied in these urban areas, where contacts between infected ticks and humans may be more frequent than elsewhere, due to high human activity. We examined the risk of encountering an infected tick in urban green spaces in Helsinki, Finland. We collected ticks at nine sites throughout Helsinki, recorded the prevalence of several pathogens and identified areas with a high potential for contacts between infected ticks and humans. Moreover, we explored the relationship between the density ofBorrelia burgdorferisensu lato-infected ticks and locally diagnosed cases of borreliosis and compared the potential for human-tick encounters in Helsinki to those in nonurban areas in south-western Finland. During 34.8 km of cloth dragging, 2,417Ixodes ricinuswere caught (402 adults, 1,399 nymphs and 616 larvae). From analysed nymphs, we found 11 distinct tick-borne pathogens, with 31.5% of nymphs carrying at least one pathogen. Tick activity was highest in August and September, leading to the density of nymphs infected withB. burgdorferis.l., and concurrently infection risk, to also be highest during this time. Nymph densities varied between the sampling sites, with obvious implications to spatial variation in infection risk. While ticks and tick-borne pathogens were found in both Helsinki and nonurban areas in south-western Finland, the estimates of human activity were generally higher in urban green spaces, leading to a higher potential for human-tick contacts therein. The presence of ticks and tick-borne pathogens and high local human activity in urban green spaces suggest that they form potential foci regarding the acquisition of tick-borne infections. Risk areas within cities should be identified and knowledge regarding urban ticks increased.

BackgroundBovine respiratory disease (BRD) continues to be great challenge in calf rearing units. The urgent need to decrease the use of antibiotics and increase animal welfare in beef production has forced us to introduce new preventive methods. Vaccinations could contribute to the solution, but the high incidence of BRD already at an early age has made it difficult to introduce suitable vaccination programs. Challenge studies have shown promising results in 3-14day old calves vaccinated with intranasal BRD vaccine, but very few field trials are available to assess the efficacy of the intranasal vaccines in field conditions. We evaluated the effect of one dose of commercial intranasal vaccination on calf mortality, daily gain, and treatment incidence for BRD in one calf rearing unit. In total, 497 calves (mean age 19days) were included in our study, 247 of which were vaccinated at the time of arrival to the unit and 250 served as negative controls (unvaccinated). Vaccinated and unvaccinated calves were situated in separate compartments until weaning. Daily gain, treatment incidence, and mortality were recorded until the calves were transported to the finishing unit, which averaged 154.5days from arrival.ResultsAverage daily gain over the complete study period was 1151.9g/day (SD 137.9) for the vaccinated calves and 1139.5g/day (SD 135.9) for the unvaccinated calves. Intranasal vaccination combined with older arrival age (17days or older) resulted in a higher daily gain (47.8g/day) compared with unvaccinated calves (coef. 0.0478, p=0.003). This association was not recorded in calves that were younger than 17days upon arrival. Intranasal vaccination was not significantly associated either with mortality (OR 0.976, p=0.968) or treatment incidence for BRD (OR 1.341, p=0.120). In total, six vaccinated calves (2.43%) and six unvaccinated calves (2.40%) died during the study period.ConclusionsVaccinating arriving calves with intranasal vaccine in the calf rearing unit did not decrease the mortality or treatment incidence for BRD, but it significantly improved the weight gain in calves transported to the unit at the age of 17days or older.

Covariance-based discovery of polymorphisms under co-selective pressure or epistasis has received considerable recent attention in population genomics. Both statistical modeling of the population level covariation of alleles across the chromosome and model-free testing of dependencies between pairs of polymorphisms have been shown to successfully uncover patterns of selection in bacterial populations. Here we introduce a model-free method, SpydrPick, whose computational efficiency enables analysis at the scale of pan-genomes of many bacteria. SpydrPick incorporates an efficient correction for population structure, which adjusts for the phylogenetic signal in the data without requiring an explicit phylogenetic tree. We also introduce a new type of visualization of the results similar to the Manhattan plots used in genome-wide association studies, which enables rapid exploration of the identified signals of co-evolution. Simulations demonstrate the usefulness of our method and give some insight to when this type of analysis is most likely to be successful. Application of the method to large population genomic datasets of two major human pathogens, Streptococcus pneumoniae and Neisseria meningitidis, revealed both previously identified and novel putative targets of co-selection related to virulence and antibiotic resistance, highlighting the potential of this approach to drive molecular discoveries, even in the absence of phenotypic data.