The Chromosome-Level Genome Assembly of European Grayling Reveals Aspects of a Unique Genome Evolution Process Within Salmonids

Salmonids represent an intriguing taxonomical group for investigating genome evolution in vertebrates due to their relatively recent last common whole genome duplication event, which occurred between 80 and 100 million years ago. Here, we report on the chromosome-level genome assembly of European grayling (Thymallus thymallus), which represents one of the earliest diverged salmonid subfamilies. To achieve this, we first generated relatively long genomic scaffolds by using a previously published draft genome assembly along with long-read sequencing data and a linkage map. We then merged those scaffolds by applying synteny evidence from the Atlantic salmon (Salmo salar) genome. Comparisons of the European grayling genome assembly to the genomes of Atlantic salmon and Northern pike (Esox lucius), the latter used as a nonduplicated outgroup, detailed aspects of the characteristic chromosome evolution process that has taken place in European grayling. While Atlantic salmon and other salmonid genomes are portrayed by the typical occurrence of numerous chromosomal fusions, European grayling chromosomes were confirmed to be fusion-free and were characterized by a relatively large proportion of paracentric and pericentric inversions. We further reported on transposable elements specific to either the European grayling or Atlantic salmon genome, on the male-specific sdY gene in the European grayling chromosome 11A, and on regions under residual tetrasomy in the homeologous European grayling chromosome pairs 9A-9B and 25A-25B. The same chromosome pairs have been observed under residual tetrasomy in Atlantic salmon and in other salmonids, suggesting that this feature has been conserved since the subfamily split.Peer reviewe

Establishing Streptomycin Epidemiological Cut-Off Values for Salmonella and Escherichia coli

This study was conducted to elucidate the accuracy of the current streptomycin epidemiological cut-off value (ECOFF) for Escherichia coli and Salmonella spp. A total of 236 Salmonella enterica and 208 E. coli isolates exhibiting MICs between 4 and 32¿mg/L were selected from 12 countries. Isolates were investigated by polymerase chain reaction for aadA, strA, and strB streptomycin resistance genes. Out of 236 Salmonella isolates, 32 (13.5%) yielded amplicons for aadA (n¿=¿23), strA (n¿=¿9), and strB (n¿=¿11). None of the 60 Salmonella isolates exhibiting MIC 4¿mg/L harbored resistance genes. Of the Salmonella isolates exhibiting MICs 8¿mg/L, 16¿mg/L, and 32¿mg/L, 1.6%, 15%, and 39%, respectively, tested positive for one or more genes. For most monitoring programs, the streptomycin ECOFF for Salmonella is wild type (WT) =32 or =16¿mg/L. A cut-off value of WT =32¿mg/L would have misclassified 13.5% of the strains as belonging to the WT population, since this proportion of strains harbored resistance genes and exhibited MICs =32¿mg/L. Out of 208 E. coli strains, 80 (38.5%) tested positive for aadA (n¿=¿69), strA (n¿=¿18), and strB (n¿=¿31). Of the E. coli isolates exhibiting MICs of 4¿mg/L, 8¿mg/L, 16¿mg/L, and 32¿mg/L, 3.6%, 17.6%, 53%, and 82.3%, respectively, harbored any of the three genes. Based on the European Committee on Antimicrobial Susceptibility Testing guidelines (ECOFF =16¿mg/L), 25% of the E. coli strains presenting MIC =16¿mg/L would have been incorrectly categorized as belonging to the WT population. The authors recommend an ECOFF value of WT =16¿mg/L for Salmonella and WT =8¿mg/L for E. coli

Multiplex PCR for detection of plasmid-mediated colistin resistance determinants, mcr-1, mcr-2, mcr-3, mcr-4 and mcr-5 for surveillance purposes

International audiencePlasmid-mediated colistin resistance mechanisms have been identified worldwide in the past years. A multiplex polymerase chain reaction (PCR) protocol for detection of all currently known transferable colistin resistance genes (mcr-1 to mcr-5, and variants) in Enterobacteriaceae was developed for surveillance or research purposes. Methods: We designed four new primer pairs to amplify mcr-1, mcr-2, mcr-3 and mcr-4 gene products and used the originally described primers for mcr-5 to obtain a stepwise separation of ca 200 bp between amplicons. The primer pairs and amplification conditions allow for single or multiple detection of all currently described mcr genes and their variants present in Enterobacteriaceae. The protocol was validated testing 49 European Escherichia coli and Salmonella isolates of animal origin. Results: Multiplex PCR results in bovine and porcine isolates from Spain, Germany, France and Italy showed full concordance with whole genome sequence data. The method was able to detect mcr-1, mcr-3 and mcr-4 as singletons or in different combinations as they were present in the test isolates. One new mcr-4 variant, mcr-4.3, was also identified. Conclusions: This method allows rapid identification of mcr-positive bacteria and overcomes the challenges of phenotypic detection of colistin resistance. The multiplex PCR should be particularly interesting in settings or laboratories with limited resources for performing genetic analysis as it provides information on the mechanism of colistin resistance without requiring genome sequencing

The rise and fall of the ancient northern pike master sex determining gene

The understanding of the evolution of variable sex determination mechanisms across taxa requires comparative studies among closely related species. Following the fate of a known master sex-determining gene, we traced the evolution of sex determination in an entire teleost order (Esociformes). We discovered that the northern pike (Esox lucius) master sex-determining gene originated from a 65 to 90 million-year-old gene duplication event and that it remained sex-linked on undifferentiated sex chromosomes for at least 56 million years in multiple species. We identified several independent species- or population-specific sex determination transitions, including a recent loss of a Y-chromosome. These findings highlight the diversity of evolutionary fates of master sex-determining genes and the importance of population demographic history in sex determination studies. We hypothesize that occasional sex reversals and genetic bottlenecks provide a non-adaptive explanation for sex determination transitions

Azithromycin resistance in Escherichia coli and Salmonella from food-producing animals and meat in Europe.

OBJECTIVES To characterize the genetic basis of azithromycin resistance in Escherichia coli and Salmonella collected within the EU harmonized antimicrobial resistance (AMR) surveillance programme in 2014-18 and the Danish AMR surveillance programme in 2016-19. METHODS WGS data of 1007 E. coli [165 azithromycin resistant (MIC > 16 mg/L)] and 269 Salmonella [29 azithromycin resistant (MIC > 16 mg/L)] were screened for acquired macrolide resistance genes and mutations in rplDV, 23S rRNA and acrB genes using ResFinder v4.0, AMRFinder Plus and custom scripts. Genotype-phenotype concordance was determined for all isolates. Transferability of mef(C)-mph(G)-carrying plasmids was assessed by conjugation experiments. RESULTS mph(A), mph(B), mef(B), erm(B) and mef(C)-mph(G) were detected in E. coli and Salmonella, whereas erm(C), erm(42), ere(A) and mph(E)-msr(E) were detected in E. coli only. The presence of macrolide resistance genes, alone or in combination, was concordant with the azithromycin-resistant phenotype in 69% of isolates. Distinct mph(A) operon structures were observed in azithromycin-susceptible (n = 50) and -resistant (n = 136) isolates. mef(C)-mph(G) were detected in porcine and bovine E. coli and in porcine Salmonella enterica serovar Derby and Salmonella enterica 1,4, [5],12:i:-, flanked downstream by ISCR2 or TnAs1 and associated with IncIγ and IncFII plasmids. CONCLUSIONS Diverse azithromycin resistance genes were detected in E. coli and Salmonella from food-producing animals and meat in Europe. Azithromycin resistance genes mef(C)-mph(G) and erm(42) appear to be emerging primarily in porcine E. coli isolates. The identification of distinct mph(A) operon structures in susceptible and resistant isolates increases the predictive power of WGS-based methods for in silico detection of azithromycin resistance in Enterobacterales

Electrophoretic variation in four french populations of domesticated rainbow trout (Salmo gairdneri)

Electrophoretic variation of 12 enzyme systems, representing 26 loci in four strains of domesticated rainbow trout (Salmo gairdneri Richardson) were studied. The variances among and within strains, estimated from observed frequencies, were compared to data of other studies of biochemical polymorphism carried out in this species. These preliminary results show that a large proportion of the variability in rainbow trout is present in the French hatchery stocks studied and that isolation of these groups for several generations has not caused a significant genetic differentiation as compared to all the domesticated strains studied.Nous avons examiné les variations électrophorétiques de 12 systèmes enzymatiques, représentant 26 locus chez 4 populations domestiques de Truite Arc-en-ciel (Salmo gairdneri Richardson). Les variabilités inter et intra-populations, estimées à partir des fréquences observées, ont été comparées aux données provenant d'autres études de polymorphisme biochimique réalisées sur la même espèce. De cette étude préliminaire, il ressort qu'une large part de la variabilité de l'espèce Truite Arc-en-ciel est présente dans les stocks piscicoles français étudiés et que l'isolement de ces groupes durant plusieurs générations ne s'est pas traduit par une différenciation génétique significative vis-à-vis de l'ensemble des populations domestiques étudiées

Genetic differences for behaviour in juveniles from two strains of brown trout suggest an effect of domestication history

Because captivity constitutes a drastic environmental change, domestication is expected to induce a rapid genetic selection for behavioural traits. In this study, we searched for genetic differences in behaviour among brown trout juveniles from two strains differing for their domestication history, i.e. an almost pure native wild Mediterranean population (W) and an Atlantic domesticated strain (D). In order to assess pure genetic differences among strains, males from the two origins were mated with Mediterranean females to produce two experimental crosses (WW and WD). The swimming activity characteristics of individual WW and WD juveniles were compared before and after the application of a stress (light switched off suddenly, followed by a 5-min period of darkness). For each of the fish observed, mating type origin (WW or WD) was unambiguously reassigned by genotyping. Behavioural responses differed between WD and WW fish. Angular velocity and the time spent immobile were greater for WW fish both before and after the short period of darkness, indicating higher reactivity. Once the light had been turned on again, mean velocity and total distance travelled were higher in WD than in WW fish. WD fish tended to recover levels of swimming activity higher than those before the dark period. This study therefore demonstrates an impact of genetic origin and domestication on swimming activity repertoire (higher reactivity in WW fish), a behavioural trait of particular importance for individual ecological performance. Owing to the contrasted domestication history of the two strains used in the comparison, we assume that the domestication level largely contributes to the behavioural changes observed

Genetic differences for behaviour in juveniles from two strains of brown trout suggest an effect of domestication history

Because captivity constitutes a drastic environmental change, domestication is expected to induce a rapid genetic selection for behavioural traits. In this study, we searched for genetic differences in behaviour among brown trout juveniles from two strains differing for their domestication history, i.e. an almost pure native wild Mediterranean population (W) and an Atlantic domesticated strain (D). In order to assess pure genetic differences among strains, males from the two origins were mated with Mediterranean females to produce two experimental crosses (WW and WD). The swimming activity characteristics of individual WW and WD juveniles were compared before and after the application of a stress (light switched off suddenly, followed by a 5-min period of darkness). For each of the fish observed, mating type origin (WW or WD) was unambiguously reassigned by genotyping. Behavioural responses differed between WD and WW fish. Angular velocity and the time spent immobile were greater for WW fish both before and after the short period of darkness, indicating higher reactivity. Once the light had been turned on again, mean velocity and total distance travelled were higher in WD than in WW fish. WD fish tended to recover levels of swimming activity higher than those before the dark period. This study therefore demonstrates an impact of genetic origin and domestication on swimming activity repertoire (higher reactivity in WW fish), a behavioural trait of particular importance for individual ecological performance. Owing to the contrasted domestication history of the two strains used in the comparison, we assume that the domestication level largely contributes to the behavioural changes observed