In the same treatment, bacteria community diversity was high in H. rufa  and the least in S. arundinaceum  with Proteobacteria, Firmicutes, and Actinobacteria as the dominant phyla. Similarly, in contaminated soil treated with cattle manure, there was a significant increase (p < 0.05) in abundance of rhizobacteria genera with Pseudomonas dominating across phytoremediation species. H. rufa was dominated by Bacillus, Fusibacter, and Rhodococcus; G. barbadense was mainly associated with Luteimonas and Mycobacterium, and T. diversifolia was inhabited by Bacillus and Luteimonas. The rhizosphere of O. longistaminata was dominated by Bacillus, Fusibacter, and Luteimonas, while S. arundinaceum was largely inhabited by Sphingomonas. These rhizobacteria genera ought to be applied in the Sudd region for bioremediation.The Sudd wetland is one of the oil-rich regions of South Sudan where environmental pollution resulting from oil extraction activities has been unprecedented. Although phytoremediation is the most feasible technique, its efficacy reduces at high TPH concentration in soil. This has made rhizoremediation the most preferred approach. Rhizoremediation involves use of a combination of phytoremediation and biostimulation. The process is catalyzed by the action of rhizobacteria. Therefore, the objective of this study is to characterize rhizobacteria communities prevalent in phytoremediation species growing in hydrocarbon-contaminated soils biostimulated with cattle manure. The treatments studied were plant species only (T1), plant species and hydrocarbons (T2), plant species and manure (T3), and plant species, manure, and hydrocarbons (T4). The rhizobacteria communities were determined using pyrosequencing of 16S rRNA. In the treatment with phytoremediation species, hydrocarbons 75 g · kg Globally, crude oil is a critical resource for national development. The world greatly depends on oil and, as a result, vast amount is used, transported, and stored . Crude ord after Nigeria and Angola in Africa [rd among the 171 oil producing countries worldwide [2 that makes up approximately 5% of the total land area of the Republic of South Sudan  [South Sudan is one of the famous crude oil producing countries falling 3n Africa  and 83rdorldwide . For exaorldwide . This poorldwide  includin000 Km2) , 11. The000 Km2) . Owing t000 Km2)  making iIn all crude oil producing countries around the world, during the process of crude oil production and secondary activities such as transportation and storage, several solids, liquids, and gaseous forms of wastes and pollutants are generated . Also, sSoil contamination with PHC is a widespread problem and has hazardous implications on both environment and human health , 18. As in situ vitrification, incineration, and electrical pyrolysis. However, the above-mentioned techniques are not largely used at present because they are associated with various shortcomings such as high cost, handling of the generated excess waste, and secondary contamination [Attempts to remediate hydrocarbon-contaminated soil are a priority in national development plans of many oil-producing countries in order to counteract the harmful effects of PHC , 24. Soumination , 26. Thimination , biorememination .Bioremediation-based rhizoremediation is a biological technology with great potential of restoring PHC contaminated soils , 29. RhiAcinetobacter, Pseudomonas, Gordonia, Rhodococcus, Immundisolibacter, Luteimonas, Alcanivorax, Marinobacter, Mycobacterium, Corynebacterium, Bacillus, Ochrobactrum, Sphingomonas, and Hahella as the most active hydrocarbon degrading genera in rhizosphere. However, the abundance of these genera in the rhizosphere of contaminated soils in the Sudd ecosystems is unknown yet; it is essential for harnessing bioremediation. Furthermore, Mackova and colleagues [Compared to other soil inhabiting microorganisms, bacteria are the most dominant and, therefore, primary microbial communities that play a fundamental role in biodegradation of PHC contaminants. The various genera utilize hydrocarbons as carbon and energy sources . The deglleagues  have sholleagues , 43. Thulleagues . Therefo−1 soil confirmed as economically appropriate [−1 soil were used. The phytoremediation plant species were (i) wild cotton (Gossypium barbadense), (ii) Sudan grass (Sorghum arundinaceum), (iii) wild rice (Oryza longistaminata), (iv) false sunflower (Tithonia diversifolia), and (v) thatching grass (Hyparrhenia rufa). These plant species are abundant in the Sudd region [A screen house pot experiment was conducted at Makerere University Agricultural Research Institute, Kabanyolo (MUARIK), from January 2018 to April 2018 (120 days). The soil used in the experiment was collected from nonhydrocarbon contaminated natural undisturbed land in the Sudd region as composite topsoil samples at a depth of 0–30 cm. Treatments included two rates of partially decomposed (1.7 : 0.6 : 0.8 NPK) cattle manure, two rates of crude oil, and five phytoremediation plant species arranged in Completely Randomised Design (CRD). Cattle manure was applied at rates of 0 and 5 g·kgropriate , while cd region  and wered region .In this study, polypropylene plastic-made pots were used. The soil was apportioned into 5 kg pots. To ensure that soil, cattle manure, and crude oil are mixed thoroughly, a metallic sheet was used as a mixing base. The mixture was returned into the pots with perforated bases to allow aeration. Each pot was labeled with the name of the respective treatment. The labeled pots were left for one week before planting. To cater for any PHC losses, pots were placed on their lids. Any water leachate was used to irrigate the respective pot. The lids were also washed after every two days and the wash water used to irrigate respective pots.oC until extraction of genomic DNA.At 120 days after planting, roots were removed from the pots. The firmly attached soil to the roots was collected from all pots (60 treatments) and transported in an ice cooler to the Biotechnology Laboratory of Makerere University Regional Centre for Crop Improvement at Kabanyolo, Wakiso district, Uganda, and stored at −80μl of Taq buffer 10×, 2.5 mM of MgCl2, 200 μmol of each deoxynucleoside triphosphate (dNTP), 20 pmol each primer, 5 μg of bovine serum albumin, 1% of formamide and 2.5U Taq polymerase , and sterile filtered Milli-Q water to a final volume of 50 μl. The PCR program was as follows: denaturing step of 94°C for 3 min, followed by 35 cycles of 1 min at 94°C, annealing for 1 min at 55°C, and elongation for 1 min at 72°C, followed by a final elongation at 72°C for 10 min.Genomic DNA was extracted from 0.25 g of the rhizosphere soil sample using DNeasy Power Soil® DNA Isolation Kit (Qiagen Company) following the manufacturer's instructions. Bacterial diversity was analysed using culture independent molecular technique16S rRNA gene PCR. The primers used in PCR reactions were 341F and 785R. A GC clamp was added to forward primer (F). These targeted approximately 300 bp of hypervariable V3 region. All PCR amplifications were performed using ThermoHybaid PCR cycler . PCR mixtures were prepared with 5 µl; dNTPs, 50 µM; genomic DNA, 30 ng/µl; and each primer, 10 pmol/µl. The concentration of MgCl2 in the reaction mixture was maintained at 1.5 mM for effective amplification. MgCl2 was a cofactor for Taq enzyme and helped in adding correct dNTPs complementary to the sequence in newly synthesizing strand by binding to dNTPs. A second PCR reaction was performed using 5 ml of the first PCR products as template under the same primers (GC clamp attached to the primer U968f) and conditions specified for the first PCR reaction. The PCR reactions were performed in duplicate, in order to obtain adequate DNA amount for electrophoresis. Amplification products were checked in 1.3% agarose gels stained with ethidium bromide (0.1 mg/ml). This was followed by storage at −20°C and then sending to the LGC Genomics Sequencing Centre in Germany. Purified PCR products were pyrosequenced using Illumina MiSeq by the LGC Genomics Sequencing Centre in Germany (http://www.support.illumina.com/downloads/bcl2fastqconversion-software-v2-19.html).The concentrations used for PCR were as follows: total mixture, 25 After sequencing, demultiplexing of all libraries for each sequencing lane was done using Illumina bcl2fastq 2.17.1.14 software (folder RAW). One to two mismatches were allowed in the barcode and read when the barcode distances between all libraries on the lane allowed for it. The sorting of reads by amplicon inline barcodes (folder RAW) was done through one mismatch that was allowed per barcode. The barcode sequence was then clipped from the sequence after sorting and reads with missing barcodes, one-sided barcodes, or conflicting barcode pairs were discarded.Clipping of sequencing adapter remnants from all reads (folder AdapterClipped) was carried out and reads with final length < 100 bases were discarded. The primer detection and clipping (folder Primer Clipped) was done by allowing three mismatches per primer; pairs of primers (Fw-Rev or Rev-Fw) were present in the sequence fragments. Whenever primer-dimers were detected, the outer primer copies were clipped from the sequence. The sequence fragments were turned into forward-reverse primer orientation after removing primer sequences.filterAndTrim” function. The trimming specifications were as follows. First, truncation length (truncLen) was set to 250 bases for both forward and reverse reads. Secondly, cutoff for maximum expected error calculated from the quality score (maxEE) was set to 3 for both forward and reverse reads for quality plots. The remaining parameters were held as default. The error rate was estimated by function “learnError.” Thirdly, a dereplication process was conducted with function “derepFastq.” All identical sequencing reads were combined into one unique sequence with a corresponding abundance equal to number of reads with that unique sequence.After primer removal and clipping of sequences, read sequences were loaded into R (version 3.6) and run through DADA2 pipeline (version 1.12) . The seqmergePairs.” As defaults in DADA2, merged sequences were only output if forward and reverse reads overlapped with a minimum of 12 bases. The merged sequences were then rearranged in an Amplicon Sequence Variant (ASV) table [makeSequenceTable” and “removeBimeraDenovo,” respectively. For taxonomic classification, recommendations of Callahan and colleagues were used [assignTaxonomy” still in DADA2 package. The ASVs with chimeras were removed from analysis using “subset_taxa” function in “Phyloseq” package (https://github.com/joey711/phyloseq).Before merging, core sample inference algorithm was applied to data . The forV) table  and cleaere used , togetheere used  using fuPhyloseq” package calculated population diversity (Simpson index), evenness (Shannon index), and richness (Chao1). To test effect of treatments on bacterial community structures, PERMANOVA analysis using “adonis” function in “vegan” package was performed. To ensure that PERMANOVA results were not affected by in-group dispersions, an analysis of multivariate homogeneity of group dispersion was conducted for different treatments using “betadisper” function in “vegan” package . Differences in bacterial community dispersion between treatments were assessed using PERMDISP, since a significant PERMANOVA result may indicate either a difference in centroids or an unequal dispersion between treatments. Multivariate analysis using nonmetric multidimensional scaling (nMDS) and principal component analysis (PCA) were used to explore hierarchical structure of bacterial community composition under effects of different treatments. These were calculated from Bray–Curtis matrices using the “metaMDS” function from “vegan” package.All statistical analyses were performed in R software (V2.15.3). To estimate coverage and sampling diversity, rarefaction curves were constructed. “n = 60). The tags were obtained with a maximum of 81,480 filtered sequences clustered to 3927 amplicon sequence variants (ASVs) of sixty samples at 3% confidence interval. Rarefaction was conducted to approximate the number of ASVs in random samples. The rarefaction curves  and T2 (S. arundinaceum), respectively  had the highest number of phyla and genera. This was followed by treatments containing plant species, manure, and hydrocarbon (T4), while plant species and hydrocarbon (T2) had the least number of phyla and genera. The most and least abundant rhizobacteria communities were noted in treatments with plant species, manure and hydrocarbon (T4) and plant species and hydrocarbon (T2), respectively. Similarly, most and least diversities were observed in T4 (ectively .p < 0.000) between plant species in all tested parameters except Simpson's index (p < 0.05) decline in abundance as shown by a drop in Chao 1 values across the five phytoremediation species. Addition of cattle manure (T4) to the treatment (plant + TPH) significantly increased (p < 0.05) bacterial abundance. In S. arundinaceum, the communities quadrupled (from 2097 to 8260); in G. barbadense, there was multiple increase (from 2781 to 9540) and a near multiple increase in O. longistaminata (from 3597 to 9168) and H. rufa , while in T. diversifolia, the communities doubled (from 4223 to 9795).The richness and diversity of bacterial communities significantly differed  in diversity in the Shannon index. However, the diversity in the Simpson index was not significant (p < 0.05) across all treatments. The most abundant phylum was Proteobacteria across all plant species with or without manure and the TPH accounting for about 41.6% of all ASVs, followed by Actinobacteria (12.7%) and Firmicutes (9.8%), of all ASVs  was attributed to addition of cattle manure. Largely, manure improves soil physicochemical properties leading to improved conditions for plant and microbial growth. Therefore, the rhizosphere became a hotspot for survival of different bacteria communities thereby accounting for the increased abundance. Moreover, cattle manure contains bacteria strains, which could have enhanced biodegradation of the TPH. In all phytoremediation plant species, there were significant differences   . Comparee manure .Bray–Curtis distance nonmetric multidimensional scaling (NMDS) revealed differences in composition of bacterial communities between hydrocarbon and nonhydrocarbon treatments. In two hydrocarbon-contaminated treatments, one was with plant species and hydrocarbon (T2) and the other with plant species, hydrocarbon, and manure (T4), where bacterial communities clustered in groups  also revealed separate clustering of bacterial communities between hydrocarbon and non-hydrocarbon-containing treatments. Irrespective of phytoremediation plant species, bacterial communities in the treatment containing plant species and hydrocarbon (T2) and one containing plant species, hydrocarbon, and manure (T4) clustered separately from one with plant species only (T1) as well as one with plant species and manure (T3) . Howeverp < 0.000) between rhizobacterial communities of phytoremediation species with and without hydrocarbon contamination. In the treatments with plant species alone (T1), high bacterial diversity was noted. However, when hydrocarbons were introduced (T2), there was a great reduction in diversity. The bacterial community shifts and eventual decrease in richness resulted from perturbations that normally occur in hydrocarbon contaminated soil. Past studies [Luteimonas, Pseudomonas, and Sphingomonas (phylum Proteobacteria), Mycobacteria and Rhodococcus (phylum Actinobacteria) and genera Bacillus and Fusibacter (phylum Firmicutes) were abundant in the treatment with plant and hydrocarbon (T2) and one with plant species, hydrocarbon, and cattle manure (T4).There were significant variations . The inclusion of cattle manure posted both direct and indirect benefits for the survival of bacterial communities. Directly, cattle manure amendments improved soil physicochemical characteristics enabling speedy adaptation by microorganisms. Furthermore, the introduction of cattle manure must have increased on soil fertility by adding soil organic carbon (SOC), total nitrogen (TN), and NPK. This must have improved plant resilience and performance in the PHC contaminated soil. Accumulation of soil organic carbon for example not only results in increased microbial biomass but also affects microbial community structure and functional diversity . TherefoIn the rhizosphere, the bacterial colonize the root surfaces, compete against other microbes and form synergestic interactions with host plants . Phyla PAssimilation is a complex biological oxidation process enhanced by supplementation with fixed nitrogen, phosphate, and other nutrients . For exaThe three phyla are also known for secreting oxygenases. In the test samples, oxygenases both monooxygenases and dioxygenases could have been secreted by the phyla. Monooxygenases catalyze desulfurization, dehalogenation, denitrification, ammonification, hydroxylation, biotransformation, and biodegradation of various aromatic and aliphatic compounds, while dioxygenases introduce molecular oxygen into their substrate . TherefoResults from NMDS and PCA showed clustering of bacteria in hydrocarbon contaminated soil with plant species (T2) and one biostimulated by manure (T4), while in nonhydrocarbon contaminated soil with plant species only (T1) and one with plant species and manure (T3), the communities of bacteria were scattered from each other. The clustering could be associated with catabolic potential of dominant bacteria phyla established by this study. Although Sutton  reasonedAddition of cattle manure to treatments with TPH increased clustering of taxa. Biostimulation with cattle manure boosted growth performance of the rhizosphere of phytoremediation plant species. The associated exudates were colonized by taxa leading to increased clustering as observed in treatments with plant species and hydrocarbon (T2) and plant species, hydrocarbon, and cattle manure (T4) see . This coBacillus, Fusibacter, Luteimonas, Mycobacterium, Pseudomonas Rhodococcus, and Sphingomonas were abundant in rhizosphere of phytoremediation species. In the same vein, the study also established that addition of cattle manure enhanced multiplication of these genera. Therefore, it is concluded that, in order to achieve better bioremediation, TPH contaminated soils should be bio-stimulated with cattle manure to increase rhizobacteria richness. With the exception of Mycobacterium (a genus that includes dangerous pathogens), this study recommends use of the genera listed above as an inoculum during ecorestoration of PHC contaminated soils in Sudd region, South Sudan. Mycobacterium is a carrier of tuberculosis (TB) which is a common cause of death with a prevalence rate of 257 per 100,000 people in South Sudan.Plant species growing in TPH contaminated soil are inhabited by various strains of rhizobacteria because their roots provide excellent living conditions. In this study, rhizobacteria genera


With the increased consumption of mixed dishes, the need for more precise quantitative data on individual food components is becoming more important. This paper aims to examine the consumption of meat, poultry, and fish before and after disaggregation of mixed dishes, and its contribution to energy and nutrient intakes in a representative sample of Australians.This study utilised a 24-h recall of 12,153 people aged two years and over participating in the 2011–12 National Nutrition and Physical Activity Survey. Consumption of meat/poultry/fish was examined before and after disaggregation of mixed dishes where all meat products and mixed dishes were separated into individual food components. Comparison between these two methods was undertaken for consumption data and contribution to energy and nutrient intakes, reported by meat type.Disaggregation of mixed dishes resulted in lower estimated intakes of red meat (9%), poultry (25%), and fish (18%) but higher estimates of processed meat (17%). Meat/poultry/fish contributed approximately 25% of total energy intake, 49% protein, 29% saturated fat, 26% iron, and 38% of zinc intake after disaggregation, which was significantly higher than their contributions reflected in survey data containing mixed dishes. Per-capita consumption of all meat/poultry/fish was 118 g/day for children and 162 g/day for adults, with chicken and beef being the highest contributors.These findings provide a detailed picture of meat/poultry/fish consumption in Australia, and emphasise the need for population studies to disaggregate reported food information to provide a more precise estimate of consumption.The online version of this article (doi:10.1186/s40795-017-0171-1) contains supplementary material, which is available to authorized users. Interest in meat consumption and its effect on health has grown tremendously over the past few years –5. In AuIn order to estimate meat consumption, detailed information is required on intakes of meat consumed as individual foods as well as recipe information for mixed dishes and meat products. For example, meat-based mixed dishes such as stir-fries and curries, typically contain meat, sauce, vegetables and/or cereals such as rice or pasta. Consumption data may be reported in broad food groupings, for example as in the first release of the national nutrition survey results , or as dIn this study, a detailed analysis of meat/poultry/fish consumption is undertaken using the 2011–12 National Nutrition and Physical Activity Survey of Australia (NNPAS), the most recent nationally representative survey and the first since 1995 covering both children and adults. All meat, poultry, fish and processed meat consumed were examined by disaggregating all meat products and mixed dishes into individual meat types and other food components. The aim of this study was to compare the impact of disaggregation on population meat/poultry/fish intake levels and nutrient contribution, as well as an assessment of the consumption of disaggregated meat/poultry/fish types according to gender, age group, and socio-economic status, factors known to affect consumption . These fThis study utilized the 2011–12 National Nutrition and Physical Activity Survey (NNPAS), undertaken by the Australian Bureau of Statistics (ABS) between May 2011 and June 2012. Ethics approval for the survey was granted by the Australian Government Department of Health and Ageing Departmental Ethics Committee in 2011 .. The interviewers used a food model booklet with sample pictures and measurements to assist the respondents with describing the amount of foods and beverages consumed [The NNPAS survey provided food and nutrient information from 24-h recalls and information on selected dietary behaviours by age group and gender at the national level. To take account of possible seasonal effects on health and nutrition characteristics, the NNPAS sample was spread across a 12-month enumeration period. The survey covered a sample of approximately 9500 private dwellings across Australia. Further details about the scope and the methodology of the survey are available from the ABS . A totalconsumed . A seconconsumed .The first release of nutrition data from the NNPAS survey reported food and nutrition intake data based on the food classification system developed by Food Standards Australia and New Zealand (FSANZ) . The sysn = 1545). For example, the proportion of chicken in a green chicken curry was determined from the AUSNUT 2011–13 recipe file, and translated into grams of chicken consumed per participant [In order to capture all meat/poultry/fish consumed on the day prior to the interview, all meat products and mixed dishes were disaggregated into individual components using the AUSNUT 2011–13 recipe file . The recticipant . Total mThe term ‘meat/poultry/fish’ as used in this study refers to all red meat, poultry, fish/seafood, organ/offal meat and processed meat but excludes any meat alternatives such as eggs, tofu, nuts and seeds, legume and beans. ‘Red meat’ refers to mammalian meat: beef , lamb (including mutton), pork, kangaroo, and game meat  excluding organ/offal meat as described in the Australian Guidelines of Healthy Eating (AGHE) . ‘PoultrThe contribution to energy and nutrient intake was calculated for total meat/poultry/fish intake and by individual categories of meat/poultry/fish for each participant and then averaged across the relevant age/gender groups. The key nutrients examined included protein, total fat, saturated fat, long-chain omega 3 fatty acids, iron and zinc. and per-consumer intakes (average intake among consumers only) of total and individual meat, poultry and fish categories. Per-capita intake was reported as mean and standard deviation (SD) expressed as gram/day. Median intake, 25th and 75th percentiles were reported for per-consumer consumption. Chi-square, ANOVA or non-parametric  tests were performed where appropriate to test for associations between meat intake and gender, age and socio-economic categories.Total meat consumption was calculated before disaggregation (using the survey classification) and after disaggregation, and per-capita intakes were compared using paired sample t-tests. Descriptive statistics were used to report the proportion consuming, per-capita P-value of <0.05 was considered statistically significant.Meat/poultry/fish consumption analyses were reported according to gender, age group as defined by NNPAS  and socio-economic categories (based on the Socio-Economic Index of Disadvantage for Areas (SEIFA), where the first SEIFA quintile indicates the most disadvantaged areas) [Table P < 0.01). The proportion of meat/poultry/fish consumption was 90.4% for children and 91.5% for adults. Red meat was consumed by 48.6% of participants, with beef as the most frequently reported type . Poultry was consumed by 37.7% of participants mainly as chicken . Fish/seafood was consumed by 21.4% of participants . Processed meat was consumed by 37.8% of participants, with higher frequencies reported by males than females . The most frequently reported types of processed meat consumed were ham , bacon , and sausage .After disaggregation of all meat products and mixed dishes, approximately 92.6% males and 90.1% females reported consuming some meat/poultry/fish on the day surveyed (Table P < 0.01), except canned fish and fish/seafood products. Adult males aged between 19 and 30 years were the highest consumers of meat/poultry/fish, and males aged 14–70 years were the highest red meat consumers , followed by beef , finfish , sausage , and lamb  (Table P < 0.01). For adults, the per-capita total meat/poultry/fish intake decreased with advancing age (P < 0.01), due mostly to a decrease in the consumption of poultry (P < 0.01).For children Fig. , the perP < 0.01) but remained unchanged for adults  and 26.4 g/day for adults  (Table P < 0.01) and females (P < 0.01) .No significant differences were observed in the per-capita intakes of fish/seafood by gender or age category. When analyzed according to SEIFA quintiles, fish/seafood intake increased with socio-economic status, for both males , followed by sausage  and finfish  (Table P < 0.01), except seafood and fish/seafood products. For children, per-consumer intake of all meat types in red meat, poultry, and fish/seafood was greater for males than females (P < 0.01). For children, per-consumer intake of processed meat types was similar between males and females, except sausage . No significant differences were observed in per-consumer intakes when analyzed by socio-economic quintiles.For adults, per-consumer intake of all meat types was greater for males than females (P < 0.01), whereas for adults this decreased with advancing age (P < 0.01).Meat/poultry/fish contributed 25.0% of total energy intake, 48.7% of protein intake, 48.9% of total fat intake, 25.8% of iron, and 37.9% of zinc intake .This secondary analysis provides nationally representative data on meat/poultry/fish consumption patterns using disaggregated data for over 1500 recipes and meat products. The use of disaggregated data reflect a more precise consumption profile of this food group and demonstrate the effect of disaggregation on survey data using broad food groupings. Our analysis revealed that the use of broad food groupings to estimate meat consumption tended to overestimate intakes of red meat by 9%, poultry 25%), and fish (18%) but underestimated intakes of processed meat (−17%). The overestimation of red meat, poultry and fish was likely due to the broad food grouping method capturing all food components (such as vegetables and grains) within mixed dishes, (for example curries and stir-fries), where meat/poultry/fish was a major component. The underestimation of processed meats is likely due to the broad food grouping method not capturing meats from mixed dishes where meat is only a minor component, such as pizzas, pies and pasta dishes. Consequently, nutrient contributions from meat/poultry/fish were also underestimated in non-disaggregated survey data, with estimates showing differences in key nutrient contributions up to 50%. These findings confirm results from previous studies , 25, 26 %, and fiBased on the disaggregated data, more than 90% of participants reported consuming meat/poultry/fish on the day prior to the interview, with red meat  consumed by approximately half, poultry and processed meat by two fifths, and fish/seafood by one fifth of the population. Per-capita data revealed total meat/poultry/fish intake was 152 g per day (118 g for children and 162 g for adults). Red meat  was the highest contributing meat category (38%), followed by poultry (30%), processed meat (17%) and fish/seafood (15%). Within the red meat category, beef was the most popular meat type, followed by lamb and pork while kangaroo and game meat were consumed in minimal amounts. In the poultry category, chicken was the major meat type, with other poultry meats such as duck, turkey and quail only contributing less than 2 %. In the fish/seafood category, finfish and canned fish were the highest contributors. Organ and offal meat consumption was negligible. In the processed meat category, sausage followed by ham and bacon contributed most to per-capita intake. Overall, the most popular meat type for children and adults of all ages, was chicken.Comparison of per-capita intakes to other surveys is problematic due to methodological differences. For example, previous analyses of Australian national nutrition surveys have not disaggregated all meat components from mixed dishes as was done in the present analysis , 28. OurRed meat consumption for adult males and females was estimated at 75 g and 62 g per day, respectively. Assuming these mean intakes are representative of daily intakes, the total red meat intake on a weekly basis can be estimated to be 525 g for males and 430 g for females. The recommendation for red meat consumption for Australian adults is set at 455 g per week based on both meeting nutrient requirements and as a limit to avoid excessive consumption associated with increased risk of colorectal cancer risk (> 700 g cooked red meat per week) .Consumption of fish/seafood was 22 g per-capita per day, translating to 154 g per week, or approximately 1.5 serves per week. This is below the recommended dietary guidelines (two serves per week)  but compConsumption of processed meat was relatively common with 38% of the population reporting consuming some type of processed meat on the day prior to the interview, although per-consumer intake was lower than that for non-processed meats (45 g versus 80–100 g). Previous findings that processed pork (including ham and bacon) is the most frequently consumed type of pork in the 2007 Australian National Children’s Nutrition and Physical Activity Survey are reflected in our analysis. Comparisons with other surveys are limited due to the differences in definition of processed meats and reporting methods , 35.Per-consumer data revealed that some meat types were reported in larger amounts per day than others, for example the quantities of lamb, sausages and finfish were larger than those of beef and pork. This is likely to reflect the different portion sizes of various meat cuts consumed, for example two sausages versus a small amount of minced pork in a stir-fry. Most processed meats (except sausage) and seafood were consumed in small quantities (20–50 g) per day.Socio-economic status was positively associated with fish/seafood consumption but not with any other meat/poultry category. Previous analysis in Australia and the US showed only very modest and inconsistent differences of total meat consumption across socio-economic categories , 29. DatThe contribution of meat/poultry/fish to intakes of key nutrients was highlighted in this secondary analysis, particularly for protein, omega-3 fatty acids, iron, and zinc. Beef, chicken, and fish contributed higher amounts of these nutrients to overall intakes when compared with processed meat. Similar findings have been indicated in the U.S. for iron and zinc intake contribution from meat/poultry/fish consumption . In contThe significant discrepancy between the contribution of meat/poultry/fish to intakes of key nutrients, in combination with the high proportion of participants consuming meat/poultry/fish, highlights the importance of recipe disaggregation. Another area that would benefit from the use of disaggregated data is the risk assessment for chemical food contaminants that may occur in meat.Of particular importance to nutrition surveys is a widely observed tendency for people to misreport their food intake. The prevalence of potential under-reporting behaviour in the NNPAS was calculated to be between 19 and 23% using the Goldberg cut-off  by the ABS . The impThe strength of this study included the use of a large representative sample of Australian children and adults. All meat categories and meat types were well described, and stratifications by gender, age, and socio-economic subgroups were undertaken to enable comparison to other studies. A potential limitation was the use of a single 24-h recall to estimate food consumption, although this is a valid method to estimate mean group intakes. The second day was not used as the individual meat types were infrequently consumed (e.g. lamb and pork) and the statistical methods analysing usual intake of such foods requires further dietary assessment on usual frequency of consumption .Our analysis on the basis of recent nationally representative data shows that red meat, poultry and fish intake is over-estimated and processed meat under-estimated compared with more precise disaggregated data. These findings emphasise the need for population studies to disaggregate reported food information to provide a more precise estimate of consumption and are of particular relevance when considering associations between foods/nutrients and health outcomes , monitorAdditional file 1: Table S1.Table S2. Mean per-capita intake of meat/poultry/fish (g) by socio-economic category after disaggregation of mixed dishes. Table S3. Daily energy and key nutrient intakes from meat/poultry/fish consumption – comparison before and after disaggregation of mixed dishes. Table S4. Proportion of persons consuming meat/poultry/fish by gender for children and adults after disaggregation of mixed dishes and per-consumer intake of meat/poultry/fish by gender for children and adults, median (25th and 75th percentile) after disaggregation of mixed dishes, g/day. (DOCX 26 kb)Categorisation of meat/poultry/fish.


Lonicera japonica is a traditional medicinal plant well known for its anti-inflammatory effect. The complete chloroplast genome sequence of L. japonica collected from Korea was obtained by de novo assembly using whole genome sequence data. The chloroplast genome is 155,060 bp in length, containing 88,853 bp in a large single copy (LSC), 18,653 bp in a small single copy (SSC) and 23,777 bp in a pair of inverted repeats (IRs). A total of 112 genes including 78 protein-coding genes and 34 structural RNA genes were identified. The sequence comparison of two L. japonica collected from Korea and China revealed 48 single nucleotide polymorphisms (SNPs) and 45 insertions/deletions (InDels). In addition, phylogenetic analysis represented intraspecific diversity within L. japonica species collected in Korea and China. Lonicera genus belongs to the Caprifoliaceae family which is close to the Rubiaceae family and consists of approximately 200 species that are mainly distributed in East Asia. Among them, about 100 inhabit in China, about 25 in Japan and about 30 in Korea. Many Lonicera species have been used as herbal medicines. L. japonica, called golden-and-silver honeysuckle or Japanese honeysuckle, has been widely used in traditional herbal medicine  has been reported  and compared it with the Chinese L. japonica chloroplast genome.The de novo using CLC genome assembler  as previously described , followed by manual confirmation using Artemis programme  was 155,060 bp in length, and separated into four distinct regions: a large single copy (LSC) , a small single copy (SSC) , and a pair of inverted repeats (IRs) . In Korea J. japonica collection chloroplast genome, a total of 112 genes including 78 protein-coding genes, 30 tRNA genes, and 4 rRNA genes were identified.Complete chloroplast genome of L. japonica collections revealed that the Korea collection was 18 bp shorter than China collection, 5 bp and 19 bp shorter in LSC and SSC regions, respectively, but 3 bp longer in each IR region. We identified 48 SNPs and 45 InDels between these two L. japonica collections that show richer intra-species diversity such as Panax ginseng and Brassica species  has a long branch and it is thought more diversified from their common ancestor than L. japonica (Korea collection).A phylogenetic tree based on complete chloroplast genome sequences was constructed with seven other species including two in Caprifoliaceae and five in Adoxaceae . As expe


Enterococcus faecium (VREfm), an increase in certain strain types marked by sequence type (ST) and cluster type (CT) has been reported in Germany over the past few years. Outbreak analyses at Charité - Universitätsmedizin Berlin revealed the frequent occurrence of VREfm ST117 CT71 isolates in 2017 and 2018. To investigate whether ST117 CT71 have emerged in recent years or whether these strains have been circulating for a longer time, we retrospectively analyzed non-outbreak strains that occurred between 2008 and 2018 to identify frequent STs and CTs.In addition to an overall rise in vancomycin-resistant In total, 120 VREfm isolates obtained from clinical and surveillance cultures from the years 2008, 2013, 2015, and 2018 were analyzed. Thirty isolates per year comprising the first 7–8 non-outbreak isolates of each quarter of the respective year were sequenced using whole genome sequencing. MLST and cgMLST were determined as well as resistance genes and virulence factors. Risk factors for VREfm ST117 were analyzed in a multivariable analysis with patient characteristics as possible confounders.p = 0.012). In 2008, vanA genotype accounted for 80% of all ST117 isolates compared to 6% in 2018. VanB CT71 first appeared in 2018 and predominated over all other ST117 at 43% (p < 0.0001).The percentage of VREfm of type ST117 increased from 17% in 2008 to 57% in 2018 , dfrG, aac(6′)-Ii, gyrA, parC and pbp5) and virulence factors  in CT71 was also found in other ST117 non-CT71 strains, mainly in CT36. The study population did not differ among the different calendar years analyzed in terms of age, gender, length of stay, or ward type (each p > 0.2).The set of resistance genes (vanB genotype in 2018. We did not detect resistance or virulence traits in CT71 that could confer survival advantage compared to other CTs among ST117 strains. To date, it is not clear why ST117 and in particular strain type ST117 CT71 predominates over other strains.This study revealed an increase in ST117 strains from 2008 to 2018, accompanied by a shift toward CT71 strains with the  Consequently a variety of resistance and virulence genotypes have emerged [In recent years, Vancomycin-resistant enterococci (VRE) have been on the rise among hospitalized patients in Germany . Infectiromosome , 6. Vanc emerged .In recent years, the German National Reference Centre for Staphylococci and Enterococci has reported the frequent occurrence of VREfm strains of sequence type ST117 as determined by multi-locus sequence typing (MLST). Of their collection of 91 isolates from blood samples in 2016, more than half were ST117 . FurtherUnpublished outbreak analyses at Charité - Universitätsmedizin Berlin (Charité) also revealed the frequent occurrence of VREfm ST117 CT71 isolates in 2017 and 2018. In order to understand the local epidemiology and strain characteristics, we investigated whether ST117  has only emerged in recent years or whether these strains have instead been circulating for a longer time and have been identified more often through the broad use of molecular typing methods.Thus, we retrospectively analyzed trends in ST and CT types among non-outbreak strains in the past decade.We retrospectively analyzed VREfm cultures, clinical and surveillance, routinely collected at Charité at five-year intervals in 2008, 2013, and 2018. The first 7–8 consecutive non-outbreak isolates per quarter from all Charité wards were included, comprising VRE positive clinical or screening cultures from individual patients. Because a recent publication suggested an increase in CT71 in Germany between 2015 and 2016 [We retrieved epidemiological data from the patient data management system and included patient age and sex, date of sampling, ward, length of stay (LOS) at the time of specimen collection, and specimen collection site.n = 76, 63%), blood cultures , urine samples , wound swabs , stool samples , nasal- and throat swabs , and other clinical cultures . Rectal swabs, throat swabs, and nasal swabs were considered screening specimens, all others were considered clinical specimens. Wards were divided into four categories: intensive care unit (ICU), hematology/oncology, surgery, and others. The latter category included standard care wards such as nephrology, cardiology and gastroenterology. Multiple assignment of one sample to different categories of ward type was not allowed.In our analysis, we included both clinical and screening cultures. Due to Data Protection Regulations, we lack the necessary data to diagnose associated infections. VREfm samples comprised rectal swabs  using multiplex PCR according to the study of Patel et al. [vanA and vanB. Vitek®2 System  or MALDI-TOF MS  were used for identification and antimicrobial susceptibility testing since 2013. Moreover, we performed an alcohol tolerance assay as described in Pidot et al. [Determination of VRE was performed using chromID® VRE agar plates  and disc diffusion method with 5 μg vancomycin and 30 μg teicoplanin from MASTDISCS®  as recommended by the European Committee on Antimicrobial Susceptibility Testing (EUCAST). In addition, we tested for the presence of l et al. . Since 2t et al.  for a spE.faecium cgMLST task template with default parameters  and E.faecium reference genome NC_017022.1 (GenBank accession number: GCA_000250945.1) was used to extract MLST and cgMLST data as described previously [https://github.com/s-andrews/FastQC), and Illumina Analysis Viewer . The ResFinder and VirulenceFinder web server (http://www.genomicepidemiology.org) was used to identify resistance genes and virulence factors, using ResFinder3.2 with the setting searching for acquired antimicrobial resistance genes as well as chromosomal mutations associated with antibiotic resistance, and VirulenceFinder2.0 (respectively threshold of 90% minimum sequence identity and 60% minimum length identity cut-off).VREfm isolates were stored as cryocultures, subcultured on blood agar and incubated overnight at 37 °C. DNA extraction was performed using the UltraClean Microbial DNA isolation kit following the manufacturer’s instructions . The quantity and purity of the DNA was measured by QuantiFluor ONE dsDNA System  and Eppendorf Biophotometer . Short read sequencing libraries were generated from genomic DNA using the Nextera XT DNA library preparation kit  and were sequenced on the MiSeq system  with 250-cycle paired-end chemistry according to the manufacturer’s instructions. Isolates were sequenced to reach 100-fold coverage. After sequencing quality-trimming, de novo assembly with the Velvet assembler and gene-by-gene comparison approach using the SeqSphere+ software version 4.1.9  were performed. For the gene-by-gene comparison, the eviously . The qua3P analysis pipeline was used [PubMLST.org database. Antibiotic resistance genes and virulence factors were analyzed with the use of CARD database [In parallel, the recently published ASAwas used . Brieflywas used  and de nwas used . Contigswas used  and annowas used . The ST database  and VFDBdatabase . Additiodatabase  was creadatabase . The Prodatabase . The resp ≤ 0.05 were entered into the model. All tests of significance were two-tailed, and p < 0.05 was considered statistically significant. All statistical analyses were performed using SPSS  and SAS .Either number and percent or median and interquartile range were calculated for descriptive analysis. Differences were tested using the Chi-square test or Wilcoxon rank-sum test. To analyze risk factors for the occurrence of ST117, we used a logistic regression model for multivariable analysis with patient age and sex, calendar year, type of ward, length of stay (LOS) at the time of specimen collection, and specimen collection site as possible confounders. Parameters with a significance of The bacterial isolates were collected in the course of routine active surveillance and infection prevention control according to guidelines of National Healthcare Authorities. Personal data were anonymized and handled in compliance with the General Data Protection Regulation and medical-ethical guidelines of Charité - Universitätsmedizin Berlin for anonymized use of patient materials. Therefore, ethical approval by an institutional board was not necessary.p = 0.839), gender , LOS , or the above mentioned ward type. Between 2008 and 2018, there was no difference in intensive care unit, hematology/oncology, surgery, or others . In contrast, LOS at the time of specimen collection and site of specimen collection differed in various calendar years. In 2008, most of the samples were obtained as clinical cultures , while in 2013 , 2015 , and 2018  screening cultures were more frequent. Urine accounted for the majority of clinical cultures. The LOS at the time of specimen collection decreased from 2008  to 2018  and is consequently of significance, p = 0.011.All VREfm isolates from the years 2008, 2013, 2015 and 2018 were analyzed and came to a total of 120 cases. We investigated patient and isolate details, including age, sex, LOS, LOS at the time of specimen collection, type of ward, and site of specimen collection. In single quarters of each year, a variety of different wards were represented. The VREfm cases did not differ among calendar years with regard to age . In total, 43 (35.8%) of the 120 E. faecium isolates were classified as ST117. In addition to ST117 strains, we detected strains assigned to ST203 (11.7%), ST80 (7.5%), ST78 (9.2%), ST192 (8.3%), ST17 (6.7%), and others (each ≤5%) . Of all 120 isolates of the years studied, CT71 (11%) was the most common CT, followed by CT36 and CT162 (both 8%), and CT164 and CT894 (both 6%).When classifying the strains with a higher resolution into cgMLST, there was a clear shift of dominant CTs from CT164 in 2008 to CT71 in 2018. We did not find any CT71 isolates in 2008, 2013, or 2015 although there was a variety of other CTs such as CT24, CT36 and CT190 , urine samples  and stool samples . Regarding the rectal swabs, there is a great increase from 2008 (n = 4) to 2013 (n = 26), whereas the number remained approximately constant in the following years (2015 n = 24 and 2018 n = 22). Urine samples accounted for 12% of the total number of samples, including 6 samples (20%) in 2008, 1 sample (3%) in 2013, 3 samples (10%) in 2015, and 4 samples (13%) in 2018. Stool samples only occurred in 2008 (n = 9) and no samples in 2013, 2015 or 2018. Most CT71 strains  were obtained from rectal swabs.Table p = 0.002). A similar association was found for urine as specimen collection site and ST117 strains . CT71 was not an independent risk factor for ST117. Because CT71 did not appear until 2018, the calendar year could not be estimated in the model.The multivariable analysis supported a strong association of ST117 with the calendar year  and efmA) as well as for trimethoprim (dfrF and dfrG) and aminoglycosides (aac(6′)-aph(2″), aph(3′)-III, ant(6)-Ia and aac(6′)-Ii) were detected in all ST117 strains. Moreover, genes for ciprofloxacin resistance  and ampicillin resistance (pbp5) were identified in all ST117 strains, which are intrinsic genes and expected to be present in all isolates. While almost all non-CT71 strains had several genes conferring resistance to aminoglycosides, we found only one such gene (aac(6′)-Ii) in CT71 strains, which is also an intrinsic gene. Some non-CT71 isolates featured resistances to chloramphenicol (cat) and tetracycline (tet(M)) which were absent in CT71 strains. Among all ST117 strains, 74% (32/43 isolates) displayed the vanB genotype and the vanA in 26% (11/43 isolates). There was a shift from vanA to vanB between 2008 and 2018, with 80% (4/5 isolates) and 67% (6/9 isolates) vanA in 2008 and 2013, to 100% (11/11 isolates) and 94% (16/17 isolates) vanB in 2015 and 2018, respectively  and sgrA to stimulate surface adhesion. Moreover, nearly all ST117 strains were characterized by the presence of ecbA , the E. faecium collagen binding protein A, as well as the enterococcal surface protein esp  to promote biofilm formation. The presence of a putative glycoside hydrolase hylEfm was detected in more than half of the ST117 strains . Occasionally, the virulence factor scm , the second collagen adhesin of E. faecium, occurred. All CT71 strains presented the same set of virulence factors: acm, esp, hylEfm, ecbA, and sgrA . This set of virulence factors was also detected in another 15 non-CT71 strains, a total of 28 isolates out of 43 (65%). These 15 strains belonged mainly to CT36 . The virulence factors of non-ST117 strains showed no striking difference from ST117 strains.All ST117 strains carried the virulence factor Regarding the alcohol tolerance assay, we tested five CT71 strains and one VREfm ATCC strain (ATCC 700221). At an isopropanol concentration of 23 and 60%, we could not detect any growth in any of the strains.n = 43). The isolates shared a core genome of 2166 genes (42 < = strains <=43), a soft-core genome of 130 genes (40 < = strains < 42), a shell genome of 1338 genes (6 < = strains < 40) and 1618 cloud genes (strains < 6). In summary, the core genome comprises 42% of the pangenome (5252 genes). The pangenome matrix (presence and absence of genes) revealed that the distribution of genes among strains was relatively similar. With regard to the accessory genome profile of the CT71 strains compared to the other CT strains, there seemed to be a set of genes more frequently present in CT71 strains than in the other strains  , dfrG, and aac(6′)-Ii) and virulence factors  of CT71 strains have already been reported in connection with high risk Enterococcus strains [In this study, different CTs of ST117 strains were identified; in 2018, however, CT71 clearly predominated. The analysis was comprised of non-outbreak isolates only, which were collected in different wards. Recent publications indicate the spread of CT71 throughout Germany in hospitals in different geographical regions without any presumed patient transfer . The rec strains , 30, 31.acm and sgrA in connection with the predominance of an outbreak strain type ST173 [acm, hylEfm, and esp as well as the efaAfm gene in CT71 strains, the latter of which contributes to cell wall adherence [Lee et al. has also reported the detection of the virulence factors pe ST173 . Surprisdherence . CT71 stThe analysis of patient characteristics did not provide an explanation for the predominance of either ST117 or CT71 strains. Only the year of specimen collection and urine as the site of specimen collection increased the chance of finding ST117.Even though urine as sample collection site is generally regarded as a clinical culture, the sampling site itself without any further clinical information did not necessarily prove that the patients had a urinary tract infection.vanB clones in different hospitals across the Rhine-Main area of Germany [In contrast to most previous publications, we aimed to investigate isolates outside of reported outbreaks. Even though certain clones predominated in different years, continuous outbreak scenarios seem unlikely because the strains were found in three geographically distinct hospital buildings without an apparent epidemiological link between the patients. There may be ongoing inter-hospital spread, which would corroborate recent findings of Falgenhauer et.al., who also detected near-identical isolates  of ST117 CT71  Germany . Raven e Germany . Inter-hvanA-plasmid that was transferred via horizontal gene transfer to already existing hospital-adapted vancomycin-susceptible E.faecium and, thus, generated new VREfm [vanB resistance in Germany [Horizontal gene transfer may facilitate the spread of VREfm through the exchange of mobile genetic elements such as transposons or plasmids, or a crossover between chromosomal and plasmid DNA through insertion elements . Zhou etew VREfm . The exc Germany .E. faecium strains with increased tolerance to handwash alcohols [Besides the ST117 strain type there are also reports of other emerging dominant strains like ST796  and ST20alcohols , whereasMarkwart et al. indicate specialist care hospitals and prevention and rehabilitation care centres as risk factors for a higher VRE proportion as well as patients older than 40 years . These fvanB-type strains [vanA to vanB genotype, which has been reported since 2015/2016 [vanB CT36 in 2015 and vanB CT71 in 2018.The general increase of VREfm in Germany was accompanied by an increase in  strains  and by a015/2016 , 35. We https://www.nrz-hygiene.de/surveillance/kiss/) the percentage of ICUs with active surveillance screening for VRE increased from 50% in 2013 to 86% in 2018. Surveillance cultures and early screening (decreased LOS at specimen collection) as opposed to clinical cultures and screening later during the patients’ stay on the ward may reveal different strains.Limitations of the study are the relatively small number of samples and the lack of information on patient factors both outside and inside of the hospital, such as comorbidities, antibiotic therapy, and admission to other health care institutions. In addition, the change in the numbers of active surveillance cultures for VRE and the days of LOS at specimen collection between 2008 and 2018 may have resulted in a systematic selection bias. The numbers of clinical and screening samples varied between the study years. Clinical cultures dominated in 2008, including isolates from patients with suspected or existing infection, and may therefore have more pathogenic properties. In 2013, 2015 and 2018 most samples were obtained from screening cultures. In general, there has been an increase in screening cultures throughout Germany. According to the German national nosocomial infection surveillance system .Additional file 2: Table S2. Resistance genes and virulence factors of 43 ST117 strains. Resistance genes: msrC: Macrolide, Lincosamide and Streptogramin B resistance; efmA, erm(B): Macrolide resistance; dfrF, dfrG: Trimethoprim resistance; aac(6′)-aph(2″), aph(3′)-III, ant(6)-Ia, aac(6′)-Ii: Aminoglycoside resistance; cat: Chloramphenicol resistance; tetM: Tetracycline resistance; gyrA, parC: Ciprofloxacin resistance; pbp5: Ampicillin resistance. Virulence factors: acm: Cell wall-anchored collagen adhesin; esp: Enterococcal surface protein; hylEfm: Putative glycoside hydrolase; ecbA: E. faecium collagen binding protein A; sgrA: surface adhesion; scm: second collagen adhesin of E. faecium.Additional file 3: Figure S1. Proportion of vanA and vanB genotype of ST117 strains from 2008 to 2018.Additional file 4: Figure S2. Phylogenetic approximately maximum-likelihood tree with all ST117 strains (n = 43) created via FastTreeMP and visualized with Microreact, coloured by CT71 (n = 13).Additional file 5: Figure S3. Results of pangenome analysis of all ST117 strains (n = 43) using roary_plots python script.


Background: Due to the frequency and severity of cold symptoms in children, and the risk of associated complications, effective treatments are urgently needed. Here we evaluated the safety profile and treatment benefits of Echinacea in children with acute cold and flu symptoms. Methods: A total of 79 children (4–12 years) were randomized to a treatment regimen of three or five times daily Echinaforce Junior tablets  for the prospective treatment of upcoming cold and flu episodes at first signs. Parents recorded respiratory symptoms daily during episodes in their child and physicians and parents subjectively rated tolerability. Results: EFJ was used to treat 130 cold episodes in 68 children and was very well tolerated by more than 96% positive physician’s ratings. EFJ-treated cold episodes lasted 7.5 days on average, with nine out of 10 episodes being fully resolved after 10 days. Five EFJ tablets daily reduced the average episode duration by up to 1.7 days (p < 0.02) in comparison to three EFJ tablets daily regimen. Effective symptom resolution finally contributed to a low antibiotic prescription rate in this study of 4.6%. Conclusions: EFJ tablets present a valuable option for the treatment of acute cold episodes in children showing a wide safety margin and increased therapeutic benefits at five tablets daily. Acute respiratory tract infections (RTIs), like the common cold, constitute the most frequent illnesses in Western civilization ,2. They Children are particularly susceptible to colds, experiencing 6–12 episodes per year compared to 2–4 annual episodes among adults ,6. ChildConventional therapeutics largely rely on antipyretics, decongestants, and cough suppressants, which are often, administered “off-label” as over-the-counter remedies. Although such medications are generally expected to rapidly improve the comfort of an ill child, no effect on illness duration has been proven ,11. MoreEchinacea purpurea), a medical plant originating in North America, and preparations thereof have a long history for the acute treatment and prevention of colds and flus [Echinacea purpurea (95% herb and 5% roots) exhibit strong inactivation activity against viruses involved in RTIs, without the risk of resistance development [Echinacea [The purple coneflower  tablets, an Echinacea extract-containing remedy that has been formulated to be suitable for use by patients below 12 years of age. The goal of this clinical study was to assess the safety and to systematically investigate whether an increased daily EFJ dose could reduce the duration and severity of cold episodes in children under real-life conditions. Evidence of effects was obtained by comparing two dosage groups (3 × 1 EFJ tablets/day vs. 5 × 1 EFJ tablets/day) to elaborate any dose response effects. No child was exposed to the risk of non-treatment and all participants were optimally supplied as stipulated by Gómez-Díaz et al. (2011) [In the present study, we tested the safety and treatment effects of Echinaforce. (2011) .Principally healthy children of 4–12 years of age were recruited and included upon receipt of informed consent signed by their parents . The following exclusion criteria applied: Age of ≥13 years or <4 years; presence of or predisposition to complicated respiratory infections at inclusion visit; participation in clinical trials within the last 30 days; intake of antimicrobial, antiviral, or immune-suppressive medication; intake of salicylic substances or broncho-vaxom; surgical interventions within the last 3 months; known diabetes mellitus, allergic rhinitis, atopy, asthma, cystic fibrosis, bronchopulmonary dysplasia, immune system diseases, degenerative disorders , metabolic or resorption disorders, liver or kidney disorders, other serious health conditions; known allergy to composite plants (chamomile or dandelion) or any compound of the test medication. Trial registration: Clinicaltrials.gov: NCT02198391.® (EFJ) tablet contains 400 mg of hydro-ethanolic extract (65% v/v) of freshly harvested Echinacea purpurea. Tinctures from the herb (drug extraction ratio 1:12) and from the roots (drug extraction ratio 1:11) are combined at a ratio of 95% to 5%, supplemented with excipients, and manufactured into EFJ tablets under GMP conditions. EFJ tablets were packaged into dark brown glass bottles containing either 40 or 60 tablets per bottle. One patient box contained three glass bottles, each sufficient for 10 days of treatment, together with a thermometer . EFJ tablets were batch released for low-dose (No. 040641A) and high-dose (No. 040641B) groups, and delivered to the study centers in multiples of randomization packages, containing four patient boxes each, all by A. Vogel AG .One Echinaforce JuniorThis study was designed as a randomized, parallel grouped, open-label multicenter clinical trial according to ICH-GCP guidelines and reported adhering to the CONSORT guidelines.The study was advertised in local newspapers. In order to increase the incentive of potentially interested parents/children to participate in this study, a prospective comparison of two different dosages of the study treatment was chosen instead of a direct comparison with no-treatment or placebo. Eligible children/parents were recruited by 10 pediatricians at eight participating pediatric practices across Switzerland. After informed consent form (ICF) was signed by at least the parents, children were randomly allocated to either the EFJ “low-dose” group (3 × 1 tablets daily) or “high-dose” group (5 × 1 tablets daily). Eligible children underwent a physical examination, and concomitant medication/disease assessment within the frame of a general anamnesis. Included children/parents received an access password and instructions on how to use individual online e-diaries  to record daily symptoms. Included children/parents obtained the investigational medicinal product (IMP), and children with acute respiratory symptoms at inclusion started treatment immediately. In all other cases, parents were asked to contact the permanently accessible study coordinator  for confirmation of cold diagnosis in their child.Each day during active cold episodes, the symptoms of running nose, stuffed nose, sneezing, cough, shivering, malaise, headache/limb pain, and sore throat were to be rated as “severe” (3), “moderate” (2), “mild” (1), “absent” (0), or “not assessable” (X). Additional variables-including sleep quality and additional care effort were rated following the same criteria and the body temperature was recorded in °C by means of an electric thermometer. For analysis, ratings of “not assessable” (X) were replaced with “mild” (1). A daily symptom score (SSc) was recorded for single symptoms, and a total daily symptom score (TSSc) was calculated by combining all symptom scores. Concomitant treatments, concomitant conditions, and incidence of adverse events (AEs) were retrieved via the study coordinator during routine contacts, from the investigator during interim visits, from close-out visits, or via assessment of e-diaries. All findings and AEs were coded according to the MedDRA (version 17.0GE). Concomitant medication/therapies were coded according to the WHO ATC (2013).At the final visit 2, returned study medication was counted to assess compliance, and the children underwent a physical investigation and final assessment of the safety and effectiveness variables. After each treatment cycle and at final visit 2, global subjective tolerability was rated by physicians and parents/children, and subjective effectiveness was rated by parents/children. Both could be rated as “very good” (1), “good” (2), “moderate” (3), or “poor” (4). Acceptance was determined by asking parents/children at visit 2 whether they would like to use the medication again. The applied methods were adapted from Taylor et al., who described a valid method for evaluating viral respiratory infections in children below 12 years old .A randomization list with a block size of 4 was prepared by d.s.h. statistical services  using the validated Rancode Professional 3.6 software . The randomization ratio was 1:1 with respect to low-dose and high-dose groups. At the study sites, the medication was distributed to each participant in accordance with this randomization list.For effectiveness analysis, sample size was calculated for an estimated difference of 1 score point in the TSSc between the two EFJ dosages, and a standard deviation of 1.5–2.5 points. A sample size of 100 children per dosage group was estimated to achieve a statistical power of 80–99% using a two-sided t-test with α = 0.05 ).The investigator assessed tolerability on final visit 2 as the primary endpoint. Kaplan–Meier analysis at episode level was performed to compare the episode duration and using Wilcoxon rank-sum test (time until patients were free of symptoms) between dosing groups. The end of an episode was defined as the day from which on all symptoms were rated “absent” (0) or “mild” (1) without re-increase over at least 4 days according to Kurugöl et al. . When ep® system (Version 9.3) and Testimate 6.5 .Baseline characteristics were compared between the two dosage groups using Mantel–Haenszel (MH) tests for ordinal variables, Wilcoxon and Fisher’s exact (FE) tests for dichotomous or categorical variables, and t-tests for continuous variables. Safety variables were analyzed in the safety collective (SAF). Analyses of effectiveness parameters were conducted in the intention-to-treat (ITT) collective and the as-treated collective (AT). The AT collective was formed from the ITT collective to adjust for the effective daily IMP intake deduced from compliance. Consequently, for AT analysis, 5 patients with significantly higher IMP intake (>160 % compliance) were transferred from the low dose to the high dose group. All statistical tests were two-sided at a 5% level of significance, and were interpreted in a descriptive-exploratory way. All statistical analyses were performed using the SASThis study was conducted from October 2014 to August 2015. A total of 79 children were recruited, enrolled, and randomized . Two outp = 0.231). Similar positive ratings were obtained in the ITT collective.p = 0.497). Of 68 safety ratings, only one (1.5%) in the higher dose group was attributed as “poor”.Parents/children confirmed the physicians’ overall positive tolerability assessment, with >99% giving ratings of “good” or “very good” in both dosage groups . Correction for the actual study medication intake (AT collective) further accentuated the treatment benefits for the higher dosage from 1.2 days to 1.7 days .Among EFJ-treated children, cold episodes lasted an average of 7.5 ± 3.5 days . Comparep = 0.005).After 10 days EFJ treatment, 102 of the total 120 cold episodes (85%) in the ITT collective resolved completely overall. Twelve episodes (19.7%) in the low-dose group and six episodes (10.2%) in the high-dose group continued with at least moderate symptoms beyond 10 days. Again, the dose response was more pronounced in the AT collective, where 12 episodes (23.5%) in the low-dose group remained unresolved after treatment compared to six episodes (8.7%) in the high-dose group . The average cumulative number of sick days per patient during the observation period were slightly reduced by 1.4 days in the ITT collective  and up to 2.9 days per patient in the AT collective, with five tablets daily in comparison to three tablets daily (p = 0.048). The dose response effect was again more pronounced in the AT collective regarding all assessed variables .EFJ-treated children in this study experienced an average of 1.9 episodes, which is below the reported average of 3.1 episodes during past seasons  without p = 0.396) and acceptance  were identified.In the ITT collective, 99% of parents/children subjectively rated the effectiveness of EFJ treatment as “good” or “very good” immediately after resolution of episodes. EFJ treatment was well accepted, as more than 82% of parents would use EFJ tablets again to treat cold symptoms in their children. No significant differences between dosage groups in terms of subjective effectiveness . None of the AEs were serious (SAE), and none showed any causal relationship to the study medication (ADR). A total of 13 out of the 20 AEs were cases of scarlet fever, chicken pox, herpes zoster, urinary tract infections, conjunctivitis, bronchitis, otitis media or tonsillitis. Three out of 20 AE were cases of gastrointestinal complaints such as vomiting, gastrointestinal pain, and celiac disease. Two out of 20 AEs were cases of cold associated complaints including fatigue and flu-like unspecific illness. One out of 20 AEs was due to feelings of dizziness and one out of 20 AEs was due to a disease of the Eustachian tuba.In 130 treatment cycles, 13 children reported 20 mostly mild and self-limiting adverse events (AEs), corresponding to one patient experiencing a single AE per 10 treated episodes. We recorded eight AEs among six children (16.7%) in the low-dose group, and 12 AEs from seven children (21.9%) in the high-dose group . Only six cold episodes (4.62%) necessitated antibiotic treatment: Three in the low-dose group and three in the high-dose group .Concomitant medication used for cold treatment-including analgesics/antipyretics, and prescribed antibiotics was reported for seven children (21.9%) in the high-dose group and eight children (22.2%) in the low-dose group  daily for up to 10 days. A total of 130 cold episodes were reported in 68 children overall during the 5.3 months of observation, encompassing a full cold and flu season.We found that EFJ tablets had an excellent safety profile for the acute treatment of RTIs. Both EFJ dosages elicited exceptionally positive tolerability ratings (“good” or “very good”) from physicians and from parents/children. This matched well with the low occurrence of AEs, of which none was related with the treatment, despite the close-meshed control instances in this trial.This study tested EFJ under real life conditions comparing dose-dependent effects of two regulatory approved dosages. In this regard, EFJ-treated cold episodes lasted, on average, 7.5 ± 3.5 days. Cold durations reported in literature are subject to variability due to heterogeneity in terms of recruitment setting, time-point of presentation, studied cold symptoms, applied treatment, and endpoint definitions. We have chosen Grüber et al.  and ThomBy day 10 of EFJ-treatment, most children in this study had fully recovered from cold symptoms . Again, Echinacea extracts may have a positive effect on the infection recurrence rate [During the study observation (5.3 months), an average of 1.9 cold episodes per patient was recorded. This is approximately 29% fewer cold episodes compared to past seasons (3.1–3.2 colds/year). Since acute colds were treated with EFJ, any preventive benefit was expected to unfold on recurrent infections and relapses only. This observation is in line with recent findings that acute treatment of respiratory infections with nce rate . Effectsnce rate ,27. For nce rate . This isnce rate .Mainly, children treated with the higher dosage exhibited a milder overall course . For exaEchinacea-treated children, as has previously been reported for adults with the same treatment [Primary care physicians in Switzerland apply antibiotics to 20.4% of respiratory tract infections, ,29, in creatment ,30.Echinacea under real life conditions using a pragmatic approach. Due to the high relevance for individual patients, society, and healthcare systems, we emphasize that the presented findings should be further confirmed by clinical studies that include a distinctly larger pediatric patient collective following a rigorous study design.For unknown reasons, we clearly fell short of the target sample size of recruited children; however, the number of recorded cold episodes was sufficient to reach acceptable statistical power. The two dosage groups significantly differed in their age and body variables by co-incidence, but had similar cold susceptibilities (as per history). Hence, the children in both treatment groups were comparable with respect to the studied indication. The authors state that this study does not represent a randomized controlled and double-blinded study, but it displays treatment benefits between two approved dosages of The data presented in this study underpins the advantageous safety profile of Echinaforce Junior tablets and provides evidence for clinical effects to treat common cold symptoms in children. An increase of the daily EFJ dose from three to five tablets seemed to shorten the average cold duration by up to 1.7 days and after 10 days resolved >90% of episodes completely. The overall need for antibiotics under the study conditions was low. Pediatricians may consider EFJ as a valuable and safe option for the acute treatment of respiratory tract infections in their young clients.


Clinical, disease activity and quality-of-life indices  were estimated. The sample consisted of 45 PsA patients with a mean age of 50.1 ± 11.5 years. The prevalence of sacroiliitis was 37.8% (n = 17), 47% of which was unilateral. The kappa coefficient was 0.64. Only 5 (29.4%) of the 17 patients with sacroiliitis on MRI had back pain. The most prevalent acute and chronic changes on MRI were, respectively, subchondral bone edema (26.7%) and enthesitis (20%), periarticular erosions (26.7%) and fat metaplasia (13.3%). CRP levels were higher among sacroiliitis patients (p = 0.028), and time of psoriasis was positively associated with chronic lesions (p = 0.006). Sacroiliitis on MRI was highly prevalent in our sample of PsA patients. Raised CRP levels were significantly associated with sacroiliitis, and longer time of psoriasis was predictive of chronic sacroiliitis lesions. Most sacroiliitis patients displayed no clinical symptoms.Sacroiliac joint involvement is one of the earliest manifestations of psoriatic arthritis (PsA). Magnetic resonance imaging (MRI) is a useful tool in the early diagnosis of axial disease due to its sensitivity for detecting acute and chronic changes associated with sacroiliitis. In this study, we evaluated the prevalence of sacroiliitis, acute and structural image changes on MRI in PsA patients and identified predictive clinical, laboratory and disease activity factors. Cross-sectional study on PsA patients submitted to MRI of the sacroiliac joints. The scans were evaluated by two blinded radiologists and the level of agreement was calculated ( According to clinical or radiographic evidence, the prevalence of axial disease is 25–70% in patients with peripheral PsA5 and 7–17% in patients without peripheral involvement6.Psoriatic arthritis (PsA) is an immune-mediated chronic inflammatory joint pathology, one of several spondyloarthropathies. Potentially destructive and progressive, it affects 20–30% of patients with cutaneous psoriasis. The clinical spectrum of PsA includes peripheral arthritis, axial disease, enthesitis or dactylitis, and skin and/or nail involvement7. Another study revealed associations between radiological sacroiliitis and clinical PsA patterns and concluded that 25% of the patients had radiological sacroiliitis . Sacroiliitis was significantly associated with peripheral joint erosion (p = 0.043), high psoriasis activity and severity scores (PASI) (p = 0.041) and early onset of PsA (p ≤ 0.001)2. However, only little has been published on MRI-detected changes of sacroiliitis in PsA patients or on factors predictive of sacroiliitis in this population8.Magnetic resonance imaging (MRI) is an excellent diagnostic tool for axial disease in spondyloarthropathies due to its sensitivity for detecting inflammatory lesions. It is currently used with the Assessment of Spondyloarthritis International Society (ASAS) classification criteria for axial spondyloarthritis, identifying the acute and chronic changes of sacroiliitis, and plays an important role in the follow-up of patients treated with immunobiological drugs. According to some authors, radiological involvement of the sacroiliac joint is one of the earliest signs of PsA, preceding clinical manifestations in approximately 30% of patientsThus, in this study we evaluated the prevalence of sacroiliitis, acute and structural image changes on MRI in patients with PsA and identified predictive clinical, laboratory and disease activity factors.Plataforma Brasil), the study protocol was approved by the research ethics committee of the HGF and filed under entry #2837133. All patients gave their written informed consent, and all study activities were conducted in accordance with the approved protocols and guidelines.In this observational, cross-sectional and quantitative study, semi-structured questionnaires were administered to patients attending the rheumatology service of the General Hospital of Fortaleza (HGF), Brazil, between August and October 2018. Submitted through an online national research database , moderate (> 3.2–5.0) and high (> 5.1). PASI (range 0–72) was calculated using GRAPPA for IOS and Android, whereas MASES (range 0–13) was determined by summing up points. BASDAI was dichotomized into inactive (< 4) and active (≥ 4), while HAQ was calculated for 8 domains, each with a range of 0–3, with higher scores indicating greater deficiency.Information was collected on sociodemographic and clinical variables, and the MRI scans were analyzed by two independent radiologists blinded to all clinical and laboratory data. The MRI scans were performed during the study period, regardless of the presence of low back pain symptoms. The sociodemographic variables included sex, age (years), level of schooling, city of origin, address, and household income. The clinical variables included the use of NSAIDs, disease-modifying anti-rheumatic drugs and immunobiological products. The patients were also scored with the disease activity score of 28 joints (DAS28)10, PASI10, the Maastricht ankylosing spondylitis enthesis score (MASES)10 and the Bath ankylosing spondylitis disease activity index (BASDAI)10. In addition, the health assessment questionnaire (HAQ)Blood drawn from the patients was submitted to determination of erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP). HLA-B27 was performed whenever available.All patients were submitted to MRI (Philips Achieva 1.5T) of the sacroiliac joints, with or without endovenous administration of paramagnetic contrast agents (gadolinium), in the oblique coronal plane  and axial plane in the following sequence: T1-weighted in turbo spin-echo (TSE) , STIR , T2-weighted with fat suppression , and enhanced T1-weighted in TSE  with fat suppression. The latter images were obtained after endovenous administration of 0.1 mmol gadobenate dimeglumine per kg of body weight. The equipment was configured to 20 cm field of view, 4 mm slice thickness and 2 excitations. Imaging lasted 7 min (STIR), 4 min (fat-suppressed T2 TSE), 6 min (T1 TSE) or 6 min (fat-suppressed enhanced T1 TSE).Sacroiliitis was classified as symmetric or asymmetric. The sacroiliac joints were evaluated for signs of involvement of the joint space, surrounding bone, bone marrow adjacent to the joint in each image sequence, enhancement, subchondral bone marrow edema, synovitis, enthesitis, capsulitis, cartilage abnormalities, periarticular erosions, fat infiltration of the subchondral bone marrow, and ankylosis. Chronic disease was suspected based on low-signal changes in T1 and T2-weighted images, periarticular erosions, subchondral bone sclerosis, joint space narrowing, bone bridges, and ankylosis. Active inflammatory disease was diagnosed based on high-signal erosions in STIR and fat-suppressed T2-weighted images, subchondral edema, synovitis, enthesitis, capsulitis and enhancement inside or adjacent to the joint.11, according to which active inflammatory lesions in sacroiliac joints are described as bone marrow edema (STIR images) or osteitis (post-gadolinium T1 images)11.In our evaluation of the MRI images we employed the definition of sacroiliitis in axial spondyloarthritis given by the ASAS/OMERACT MRI group12. The strength of the observed associations was expressed as prevalence ratios and their respective confidence intervals (95% CI). The level of statistical significance was set at 5% (p < 0.05).All analyses were performed with the software IBM SPSS Statistics v. 23.0. The data and results were managed with Excel 2018 spreadsheets and graphs. Qualitative variables were expressed as absolute and relative frequencies, while quantitative variables were expressed as mean values ± standard deviation or median values (1st–3rd interquartile range). The normality of the quantitative data was verified with the Shapiro–Wilk test, while the binomial test was used to evaluate differences in the prevalence of sacroiliitis between our study and the literature. In the bivariate analysis, quantitative variables were compared using the Mann–Whitney test, while qualitative variables were compared with the chi-squared test or Fisher’s exact test. We also calculated the percentage variation between the groups (duration of arthritis and psoriasis with acute and chronic changes in sacroiliitis). The inter-examiner agreement was determined by calculating kappa coefficients The sample consisted of 45 PsA patients aged 50.1 ± 11.5 years on the average who met the inclusion criteria. The two sexes were nearly equally represented , the vast majority were classified as non-white (95.6%), the predominant monthly income bracket (53.3%) was 0–3 minimum wages , and the average time of schooling was 9.5 ± 3.8 years.  for symmetric polyarticular arthritis, 22% (n = 10) for asymmetric oligoarthritis, and 22.2% (n = 10) for spondylitis. Among other clinical manifestations, the most prevalent was nail involvement , followed by enthesitis , dactylitis  and uveitis  , PASI (2.00 ± 2.9) and MASES (1.90 ± 3.2) scores  used NSAIDs, 35.6% (n = 16) received immunobiologicals, and 8.9% (n = 4) used corticoids. Historically, 73.3% (n = 33) had never used corticoids.In our sample of 45 PsA patients, the prevalence of sacroiliitis on MRI was 37.8% n = 17), 47.0% (n = 8) of which was unilateral. Only one fifth  presented symptoms of inflammatory lumbar pain. Low back pain was observed in 29.4% (n = 5) of the patients with sacroiliitis. The most prevalent acute structural change observed on MRI was subchondral bone edema , followed by enthesitis , capsulitis  and synovitis  . The corresponding value for psoriasis patients was 6–17 years (median 9). A significant association was observed between time of psoriasis and acute and chronic changes of sacroiliitis , can help prevent loss of quality of life from limited physical function, psychological distress and inability to perform daily activities due to pain and discomfort associated with joint and skin symptoms17. Loss of quality of life can also be prevented with emerging biological therapy18.To our knowledge, this is the first Brazilian study to evaluate the prevalence of sacroiliitis on MRI in PsA patients and correlate positivity for sacroiliitis with acute and chronic changes and clinical, laboratory and epidemiological variables. The joint analysis of these variables made it possible to identify predictors of sacroiliitis in our sample. Early diagnosis of axial involvement on MRI, prior to the emergence of structural damage . A similar discrepancy was observed in relation to a recent study from the US involving 1,567 patients of which only 9% were non-white20. This may be explained by differences in sample size or by regional differences . The broad demographic spectrum of the Brazilian population is the most likely explanation for the particularly non-white ethnic composition of our sample.Sociodemographically, our sample was similar to that of another Brazilian cohort of 175 PsA patients2, but is somewhat higher than the prevalence of sacroiliitis in PsA patients reported in another Brazilian study (24.4%)19. The higher prevalence observed in the current study may be explained by our recruitment from a referral center for patients with more advanced and long-standing disease, or by regional, genetic and environmental factors.The prevalence of sacroiliitis on MRI in our sample of PsA patients (37.8%) matches studies reporting prevalences of axial disease in PsA between 25 and 50%23. In fact, another study documented clinical features of sacroiliitis in 14 (33%) of 42 patients with normal MRI scans and in 10 (38%) of 26 patients with abnormal scans . Moreover, the presence of sacroiliitis on MRI was associated with restricted spinal movements (p = 0.004) and the duration of PsA (p = 0.04), but no association between sacroiliitis on MRI and BASDAI or BASFI or HLA-B27 was observed, leading the authors to conclude that clinical assessment of sacroiliitis and HLA-B27 are poor predictors of sacroiliitis diagnosed by MRI in PsA patients24. Finally, a study using MRI reported inflammatory and chronic changes to defined areas in the sacroiliac joints in patients with early-stage versus late-stage PsA and concluded that whereas the iliac and the sacral side of the sacroiliac joints are almost equally affected, the dorsocaudal synovial part of the joint is involved significantly more often than the ventral part, especially in early disease. Sacroiliac enthesitis is not a special feature of early sacroiliac inflammation25.Interestingly, inflammatory lumbar pain was only observed in 29.4% of the patients with sacroiliitis and the most prevalent findings on MRI were subchondral bone edema and enthesitis (acute changes) and periarticular erosions and fat metaplasia (chronic changes), proving that changes on MRI may occur in the absence of clinical symptoms, and reinforcing the role of MRI in the prevention of distant structural damage and in the evaluation of therapy for axial spondyloarthritis. Despite the regional and ethnic peculiarities of our sample, our results are supported by the literature showing that patients with sacroiliitis do in fact display early structural damage in the sacroiliac joints on MRI before the emergence of clinical symptomsp = 0.006), matching the results of another Brazilian study reporting a mean psoriasis duration of 15.4 ± 11.7 years26. In a controlled study involving patients with cutaneous psoriasis but no arthritis or spondylitis, the evidence of subclinical axial disease with sacroiliac involvement on MRI was limited. The authors admitted their findings were insufficient to warrant routine MRI screening of patients with long-standing psoriasis27.In our sample, the time of psoriasis (12.5 ± 9.6 years) was significantly associated with the presence of chronic sacroiliitis damage , in disagreement with the literature. Thus, two studies evaluating the type and frequency of sacroiliitis in psoriasis patients found, respectively, that 26% of 133 psoriasis patients presented sacroiliac involvement, more than half of which was bilateralp = 0.003), contrasting with our findings 2.Moreover, the presence of sacroiliitis was not significantly associated with oligoarthritis, mutilating arthritis, dactylitis, nail involvement or enthesitis. However, an earlier study found a higher prevalence of uveitis in patients with than without sacroiliitis . An earlier study2 also found significant correlations between CRP and sacroiliitis in PsA patients (p = 0.006), but no other disease activity index was significantly associated with sacroiliitis . The same was true for the quality-of-life index (HAQ p = 0.420) and clinical scores . Likewise, current PASI values were non-significant (p = 0.495), matching the literature (p = 0.23)2.In this study, the increased CRP levels resulting from disease activity were significantly associated with the presence of sacroiliitis  and early-onset arthritis2. On the other hand, the risk factors for sacroiliac involvement also include severe psoriasis associated with HLA-B0801 positivity and reduced HLA-B27 frequency, suggesting HLA-B27 is a less important predictor of sacroiliitis. In our study, sacroiliitis was not associated with ESR (p = 0.494) or with clinical predictors such as joint involvement pattern (p > 0.05) and enthesitis (p = 0.848). A similar lack of correlation between sacroiliitis and enthesitis was reported in another recent study30.Approximately 15% of patients with peripheral PsA may be expected to develop axial disease in the course of 10 years. The possible risk factors include severe peripheral arthritis, early-onset arthritis, increased ESR, presence of HLA-B27, severe ungual dystrophy and inflammatory bowel diseaseOur study has some limitations: (i) the relatively small sample of patients recruited from a single center, (ii) the unavailability of HLA-B27 testing in some cases, (iii) the cross-sectional study design, making it more difficult to draw conclusions regarding the ability of the selected clinical and laboratory variables to predict sacroiliitis in PsA patients, and (iv) the absence of analysis of the possible impact of racial type on PsA diagnosis, therapy and outcome.Sacroiliitis on MRI was highly prevalent in our sample of PsA patients, but most sacroiliitis patients displayed no clinical symptoms. Raised CRP levels were significantly associated with sacroiliitis, and the time of psoriasis was predictive of chronic sacroiliitis lesions. Prospective studies on larger samples, including HLA-B27 testing, are necessary to identify predictive clinical, laboratory and image changes on MRI in PsA patients.