Morphological, phytochemical and molecular characterization of five common Jatropha species in the Niger Delta Region of Nigeria

The economic and medicinal important genus Jatropha contains many distinctly different species. To elucidate the genetic relationship of five common occurring Jatropha species namely J. multifida, J. podagrica, J.tanjorernsis, J. curcas, and J. gossypifolia, thirty-nine morphological, six phytochemical features and one arbitrary marker was used to screen and explore their similarity. Morphological data was obtained from the measurement of vegetative and reproductive parts while the presence of five phytochemicals was determined using differentphytochemical tests. The DNA of all five Jatropha species were amplified and sequenced using Ribolose 1, 5- biphosphate carboxylase molecular marker. The DNA sequences were then aligned using the Basic Local Alignment Search Tool for nucleotide 2.8.0 version of the National Center for Biotechnology Information database and phylogenetic trees were constructed using Paleontological Statistical software and Molecular Evolutionary Genetics Analysis version 7.0.26 software. From the results of the classical and phylogenetic cluster analysis, the five Jatropha species was separated into two major clusters. The highly distinctive J. gossypifolia was the only species that clustered separately from the other Jatropha species. Although, J. tanjorensis has been reported to be a hybrid from J. curcas and J. gossypifolia, the species did not segregate and cluster with these species, but segregated with J. multifida, and J.podagrica, indicating that this species is more closely related to J. multifida, and J. podagrica than J. curcas and J.gossypifolia. The result therefore provide information that would be useful in the plant improvement programs for the genus Jatropha.

The Evolutionary History of Vertebrate Adhesion GPCRs and Its Implication on Their Classification

Adhesion G protein-coupled receptors (aGPCRs) form a structurally separate class of GPCRs with an unresolved evolutionary history and classification. Based on phylogenetic relations of human aGPCRs, nine families (A–G, L, V) were distinguished. Taking advantage of available genome data, we determined the aGPCR repertoires in all vertebrate classes. Although most aGPCR families show a high numerical stability in vertebrate genomes, the full repertoire of family E, F, and G members appeared only after the fish–tetrapod split. We did not find any evidence for new aGPCR families in vertebrates which are not present in the human genome. Based on ortholog sequence alignments, selection analysis clearly indicated two types of tetrapod aGPCRs: (i) aGPCR under strong purifying selection in tetrapod evolution (families A, B, D, L, V); and (ii) aGPCR with signatures of positive selection in some tetrapod linages (families C, E, G, F). The alignments of aGPCRs also allowed for a revised definition of reference positions within the seven-transmembrane-helix domain (relative position numbering scheme). Based on our phylogenetic cluster analysis, we suggest a revised nomenclature of aGPCRs including their transcript variants. Herein, the former families E and L are combined to one family (L) and GPR128/ADGRG7 forms a separate family (E). Furthermore, our analyses provide valuable information about the (patho)physiological relevance of individual aGPCR members.

#65: Characterization of Resistance Patterns Among Isolates of E. coli and Klebsiella sp. from Wastewater, and Clinical Samples in a Tertiary Care Children’s Hospital in Lima, Peru

Background Environmental monitoring of enterobacteria in hospital wastewater could be a useful tool to understand the composition of the microbiota in patients, their visits, and healthcare personnel, but it also may be useful for monitoring antimicrobial resistance among healthcare-associated infections in hospitalized patients. The aim of this study was to characterize and describe the phenotypic and genotypic antimicrobial resistance of Escherichia coli and Klebsiella sp. strains from wastewater and from clinical isolates in a tertiary care children’s hospital in Lima, Perú. Methods We systematically collected 70 isolates of Escherichia coli and Klebsiella sp. from wastewater and 24 isolates of the same enterobacteria from blood and urine cultures at Instituto Nacional de Salud del Niño San Borja (INSN-SB) in Lima from December 2018 to May 2019. Susceptibility profiles were evaluated following CLSI criteria. We used the Jarlier method for the detection of extended-spectrum beta-lactamases (ESBL). For detection of AmpC beta-lactamases, we used discs of cefoxitin, ceftazidime, and ceftriaxone combined with cloxacillin and cefotaxime. For the detection of carbapenemase production, we used EDTA and phenylboronic acid inhibitors between meropenem and imipenem disks. We performed genomic sequencing for the detection of resistance genes and to perform a phylogenetic cluster analysis. Results We collected a total of 94 isolates (70 from wastewater and 24 from clinical samples). Among the total isolates, 19 (20.2%) were ESBL producers. The frequency of ESBL producers in wastewater was 7.1% (5/24), whereas the frequency of ESBL producers in clinical samples was 58.3% (14/70). The most frequent resistance genes were tet (variant 34 and A) and blaTEM-1. The frequency of tet(34) and tet(A) were 10% and 7%, respectively, in isolates from wastewater, whereas the frequency of these genes were 0% and 6%, respectively, in clinical isolates. The gene blaCTX-M-15 was present in isolates from wastewater (1%) and clinical samples (2%). Phylogenetic cluster analysis found no similarities between isolates from wastewater and to those from clinical samples, suggesting that the population of these enterobacteria were different in wastewater compared with clinical samples. Conclusions Enterobacteria from hospital wastewater may not reflect the profile of infections caused by these microorganisms. However, they may reflect the microbiological microbiota among patients, their visits, and hospital healthcare personnel. Further studies that compare the phenotypic and genotypic characteristics among isolates from wastewater and the enteric microbiota from these individuals would be necessary to assess this hypothesis.