Diversity and antimicrobial resistance of Salmonella enterica serovars in surface river water and sediment

This study aimed to evaluate the contamination by Salmonella sp. in the Capinzal River, to determine the prevalent serovars, patterns of antimicrobial resistance, and the genetic relationships between the serovars identified. A total of 108 samples were collected from 2016 to 2018. The isolation of Salmonella spp. was conducted according to International Organization for Standardization (ISO) standards. The antimicrobial resistance profile of the Salmonella isolates was evaluated, and isolates were selected for serotyping and verification of genetic similarity using the Pulsed-Field Gel Electrophoresis (PFGE) Technique. Of the 108 samples collected, 35 (32.4%) were positive for Salmonella; 17.2% of the isolates were from the rural area; and 88.6% were from the urban area. Salmonella was isolated from all collect points along the river, with a higher incidence at the beginning of the urban area, indicating that contamination starts in the rural area and intensifies in the urban area of the city. A percentage of 35.1% of the Salmonella isolates were resistant to at least two antibiotics, while 18.9% were considered multidrug-resistant (resistant to at least two antibiotics of different classes). Seven serovars were distinguished from the serotyped isolates, with a prevalence rate of 23.5% for S. Infantis, S. Orion, and S. Javiana; 11.8% for S. Senfterberg, and 5.9% for S. Montevideo, S. Heidelberg, and S. enterica subsp. enterica (O: 6.8). The variability in specific restriction sites generated by PFGE resulted in 10 pulsotypes, separating mainly different serotypes.

Signal-based optical map alignment

In genomics, optical mapping technology provides long-range contiguity information to improve genome sequence assemblies and detect structural variation. Originally a laborious manual process, Bionano Genomics platforms now offer high-throughput, automated optical mapping based on chips packed with nanochannels through which unwound DNA is guided and the fluorescent DNA backbone and specific restriction sites are recorded. Although the raw image data obtained is of high quality, the processing and assembly software accompanying the platforms is closed source and does not seem to make full use of data, labeling approximately half of the measured signals as unusable. Here we introduce two new software tools, independent of Bionano Genomics software, to extract and process molecules from raw images (OptiScan) and to perform molecule-to-molecule and molecule-to-reference alignments using a novel signal-based approach (OptiMap). We demonstrate that the molecules detected by OptiScan can yield better assemblies, and that the approach taken by OptiMap results in higher use of molecules from the raw data. These tools lay the foundation for a suite of open-source methods to process and analyze high-throughput optical mapping data. The Python implementations of the OptiTools are publicly available through http://www.bif.wur.nl/.

High Level of Heterozygous Haplotype of Hemoglobin in Abidjan

Background. Sickle cell disease (SCD) is an hemoglobin disorder that concerns 300,000 newborns each year around the world. Hemoglobin haplotypes can modulate SCD clinical expression. In Côte d’Ivoire, no study has yet investigated the distribution of hemoglobin haplotypes in the population. The goal of this study was to identify hemoglobin haplotypes for people attending dispensary with mild malaria in Abidjan (Ivory Coast) independently of their SCD status. Methods To determine haplotypes, specific restriction enzyme (RE) method is used after PCR amplification with different couples of primers. According to the digestion profile of PCR products, five hemoglobin haplotypes are found over the world. ResultsIn Abidjan, four different “classical” haplotypes of hemoglobin were detected: Benin (56.5%), Bantou (28.5%), Senegal (4%), Cameroun (1%). In parallel 10% of atypical profiles were described. Heterozygous haplotype (69%) was more frequent than homozygous haplotype (31%). ConclusionsBenin haplotype usually associated with more severe of SCD symptoms was predominant in the study population. However this preliminary study highlights also a high prevalence of atypical and heterozygous haplotypes in the population.

The benefit of the diffusion kurtosis imaging in presurgical evaluation in patients with focal MR-negative epilepsy

The effectivity of diffusion-weighted MRI methods in detecting the epileptogenic zone (EZ) was tested. Patients with refractory epilepsy (N=25) who subsequently underwent resective surgery were recruited. First, the extent of white matter (WM) asymmetry from mean kurtosis (MK) was calculated in order to detect the lobe with the strongest impairment. Second, a newly developed metric was used, reflecting a selection of brain areas with concurrently increased mean Diffusivity, reduced fractional Anisotropy, and reduced mean Kurtosis (iDrArK). A two-step EZ detection was performed as (1) lobe-specific detection, (2) iDrArK voxel-wise detection (with a possible lobe-specific restriction if the result of the first step was significant in a given subject). The method results were compared with the surgery resection zones. From the whole cohort (N=25), the numbers of patients with significant results were: 10 patients in lobe detection and 9 patients in EZ detection. From these subsets of patients with significant results, the impaired lobe was successfully detected with 100% accuracy; the EZ was successfully detected with 89% accuracy. The detection of the EZ using iDrArK was substantially more successful when compared with solo diffusional parameters (or their pairwise combinations). For a subgroup with significant results from step one (N=10), iDrArK without lobe restriction achieved 37.5% accuracy; lobe-restricted iDrArK achieved 100% accuracy. The study shows the plausibility of MK for detecting widespread WM changes and the benefit of combining different diffusional voxel-wise parameters.

Signal-based optical map alignment

In genomics, optical mapping technology provides long-range contiguity information to improve genome sequence assemblies and detect structural variation. Originally a laborious manual process, Bionano Genomics platforms now offer high-throughput, automated optical mapping based on chips packed with nanochannels through which unwound DNA is guided and the fluorescent DNA backbone and specific restriction sites are recorded. Although the raw image data obtained is of high quality, the processing and assembly software accompanying the platforms is closed source and does not seem to make full use of data, labeling approximately half of the measured signals as unusable. Here we introduce two new software tools, independent of Bionano Genomics software, to extract and process molecules from raw images (OptiScan) and to perform molecule-to-molecule and molecule-to-reference alignments using a novel signal-based approach (OptiMap). We demonstrate that the molecules detected by OptiScan can yield better assemblies, and that the approach taken by OptiMap results in higher use of molecules from the raw data. These tools lay the foundation for a suite of open-source methods to process and analyze high-throughput optical mapping data. The Python implementations of the OptiTools are publicly available through http://www.bif.wur.nl/.

Rapture facilitates inexpensive and high-throughput parent-based tagging in salmonids

Accurate methods for tracking individuals are crucial to the success of fisheries and aquaculture management. Management of migratory salmonid populations, which are important for the health of many economies, ecosystems, and indigenous cultures, is particularly dependent on data gathered from tagged fish. However, the physical tagging methods currently used have many challenges including cost, variable marker retention, and information limited to tagged individuals. Genetic tracking methods combat many of the problems associated with physical tags, but have their own challenges including high cost, potentially difficult marker design, and incompatibility of markers across species. Here we show the feasibility of a new genotyping method for parent-based tagging (PBT), where individuals are tracked through the inherent genetic relationships with their parents. We found that Rapture sequencing, a combination of restriction-site associated DNA and capture sequencing, provides sufficient data for parentage assignment. Additionally, the same capture bait set, which targets specific restriction-site associated DNA loci, can be used for both Rainbow Trout Oncorhynchus mykiss and Chinook Salmon Oncorhynchus tshawytscha. We input 248 single nucleotide polymorphisms from 1,121 samples to parentage assignment software and compared parent-offspring relationships of the spawning pairs recorded in a hatchery. Interestingly, our results suggest sperm contamination during hatchery spawning occurred in the production of 14% of offspring, further confirming the need for genetic tagging in accurately tracking individuals. PBT with Rapture successfully assigned progeny to parents with a 98.86% accuracy with sufficient genetic data. Cost for this pilot study was approximately $3 USD per sample. As costs vary based on the number of markers used and individuals sequenced, we expect that when implemented at a large-scale, per sample costs could be further decreased. We conclude that Rapture PBT provides a cost-effective and accurate alternative to the physical coded wire tags, and other genetic-based methods.

Rapid generation of rotavirus single-gene reassortants by means of eleven plasmid-only based reverse genetics

Reassortment is an important mechanism in the evolution of group A rotaviruses (RVAs), yielding viruses with novel genetic and phenotypic traits. The classical methods for generating RVA reassortants with the desired genetic combinations are laborious and time-consuming because of the screening and selection processes required to isolate a desired reassortant. Taking advantage of a recently developed RVA reverse genetics system based on just 11 cloned cDNAs encoding the RVA genome (11 plasmid-only system), we prepared a panel of simian SA11-L2 virus-based single-gene reassortants, each containing 1 segment derived from human KU virus of the G1P[8] genotype. It was shown that there was no gene-specific restriction of the reassortment potential. In addition to these 11 single-gene reassortants, a triple-gene reassortant with KU-derived core-encoding VP1–3 gene segments with the SA11-L2 genetic background, which make up a virion composed of the KU-based core, and SA11-L2-based intermediate and outer layers, could also be prepared with the 11 plasmid-only system. Finally, for possible clinical application of this system, we generated a series of VP7 reassortants representing all the major human RVA G genotypes (G1–4, G9 and G12) efficiently. The preparation of each of these single-gene reassortants was achieved within just 2 weeks. Our results demonstrate that the 11 plasmid-only system allows the rapid and reliable generation of RVA single-gene reassortants, which will be useful for basic research and clinical applications.

EFFECT OF THE SUN ON EARTHʼS SEISMICITY

For the experimental study of the relationship between earthquakes and solar activity, we introduce the idea of the statistical sum Z of earthquake ensemble. A number of numerical parameters of seismicity (average planetary magnitude, entropy, fluctuations of magnitude) are expressed through Z. A severe restriction is, however, imposed on the magnitude during the formation of the ensemble. We propose an alternative method. It does not have this specific restriction, although it allows us to calculate only one numerical parameter, namely the global daily magnitude Mg. Over the 20-year period from 1980 to 1999, 7300 values of Mg have been calculated. The comparison between the Mg values and Wolf numbers W made it possible to determine the effect of the Sun on earthquakes at a statistically significant level.

EFFECT OF THE SUN ON EARTHʼS SEISMICITY

For the experimental study of the relationship between earthquakes and solar activity, we introduce the idea of the statistical sum Z of earthquake ensemble. A number of numerical parameters of seismicity (average planetary magnitude, entropy, fluctuations of magnitude) are expressed through Z. A severe restriction is, however, imposed on the magnitude during the formation of the ensemble. We propose an alternative method. It does not have this specific restriction, although it allows us to calculate only one numerical parameter, namely the global daily magnitude Mg. Over the 20-year period from 1980 to 1999, 7300 values of Mg have been calculated. The comparison between the Mg values and Wolf numbers W made it possible to determine the effect of the Sun on earthquakes at a statistically significant level.