Genomic Profiling of Multidrug-Resistant Swine Escherichia coli and Clonal Relationship to Human Isolates in Peru.

The misuse of antibiotics is accelerating antimicrobial resistance (AMR) in Escherichia coli isolated from farm animals. The genomes of ten multidrug-resistant (MDR) E. coli isolates from pigs were analyzed to determine their sequence types, serotypes, virulence, and AMR genes (ARGs). Additionally, the relationship was evaluated adding all the available genomes of Peruvian E. coli from humans using the cgMLST + HierCC scheme. Two aEPEC O186:H11-ST29 were identified, of which H11 and ST29 are reported in aEPEC isolates from different sources. An isolate ETEC-O149:H10-ST100 was identified, considered a high-risk clone that is frequently reported in different countries as a cause of diarrhea in piglets. One ExPEC O101:H11-ST167 was identified, for which ST167 is an international high-risk clone related to urinary infections in humans. We identified many ARGs, including extended-spectrum ß-lactamase genes, and one ETEC harboring the mcr-1 gene. CgMLST + HierCC analysis differentiated three clusters, and in two, the human isolates were grouped with those of swine in the same cluster. We observed that Peruvian swine MDR E. coli cluster with Peruvian E. coli isolates from healthy humans and from clinical cases, which is of great public health concern and evidence that AMR surveillance should be strengthened based on the One Health approach.

Genetic Analysis of Infectious Bronchitis Virus S1 Gene Reveals Novel Amino Acid Changes in the GI-16 Lineage in Peru.

Infectious bronchitis is a highly contagious viral disease that represents an economic threat for poultry despite the wide use of vaccination. To characterize the virus circulating in Peru, we analyzed 200 samples, including nasopharyngeal swabs and multiple tissues collected from animals suspected of being infected with infectious bronchitis virus (IBV) between January and August in 2015. All animals had at least one positive sample for IBV by RT-PCR. Out of these positive samples, eighteen (18) were selected for viral isolation and a partial S1 sequencing. Phylogenetic analysis showed that sixteen isolates clustered with members of GI-16 lineage, also known as Q1, with nucleotide homology ranging from 93% to 98%. The two remaining isolates grouped with members of the GI-1 lineage. Our study reveals circulation of GI-16 lineage during this period in poultry systems in Peru, along with GI-1 lineage (vaccine-derived). Moreover, those IBV GI-16 isolates showed unique nucleotide and amino acid changes compared to their closest relatives. Altogether, these findings reveal the circulation of GI-16 lineage while describing changes at key regions of the S protein that might be of relevance for vaccine evasion. These results highlight the importance of genetic surveillance for improving vaccination strategies against infectious bronchitis.

Draft Genome Sequences of 10 Multidrug-Resistant Escherichia coli Strains Isolated from Piglets in Peru.

Pathogenic Escherichia coli frequently causes diarrhea on pig farms. Here, we report the draft genome sequences of 10 strains of multidrug-resistant E. coli isolated from piglets.

Phylogenomics reveals multiple introductions and early spread of SARS-CoV-2 into Peru.

Peru has become one of the countries with the highest mortality rates from the current coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). To investigate early transmission events and the genomic diversity of SARS-CoV-2 isolates circulating in Peru in the early COVID-19 pandemic, we analyzed 3472 viral genomes, of which 149 were from Peru. Phylogenomic analysis revealed multiple and independent introductions of the virus likely from Europe and Asia and a high diversity of genetic lineages circulating in Peru. In addition, we found evidence for community-driven transmission of SARS-CoV-2 as suggested by clusters of related viruses found in patients living in different regions of Peru.

Diarrheagenic Escherichia coli isolates from neonatal alpacas mainly display F17 fimbriae adhesion gene.

Neonatal domestic South American Camelid llamas and alpacas suffer from an enteric disease complex characterized by abdominal distention, lethargy, dehydration, and eventual fatal septicemia. Analysis of rectal swabs from neonatal alpacas suffering clinical diarrheas has constantly isolated Escherichia coli, mainly the EPEC and EHEC pathotypes. The present communication reports the results of molecular analysis of 226 E. coli strains from neonatal alpaca rectal swabs. The isolates were initially tested by multiple PCR, to identify E. coli virulence genes eae, bfp, Lt, Stx1, Stx2, sta, stab, and lt genes and a similar test to detect F4, F5, F6, F17, and F41fimbriae adhesin genes. Forty-two of the 226 (18.5%) isolates tested positive for at least one pathogenic gene, 25 of the 42 were classified as EPEC (3 positives for only eae and 22 for both eae and bfp) and the remaining 17 were classified as EHEC. Twenty-four (57%) of the 42 isolates tested positive to F17 adhesin while one was positive for both F6 and F17.

Pathogenomics insights for understanding Pasteurella multocida adaptation.

Pasteurella multocida is an important veterinary pathogen able to infect a wide range of animals in a broad spectrum of diseases. P. multocida is a complex microorganism in relation to its genomic flexibility, host adaptation and pathogenesis. Epidemiological analysis based on multilocus sequence typing, serotyping, genotyping, association with virulence genes and single nucleotide polymorphisms (SNPs), enables assessment of intraspecies diversity, phylogenetic and strain-specific relationships associated with host predilection or disease. A high number of sequenced genomes provides us a more accurate genomic and epidemiological interpretation to determine whether certain lineages can infect a host or produce disease. Comparative genomic analysis and pan-genomic approaches have revealed a flexible genome for hosting mobile genetic elements (MGEs) and therefore significant variation in gene content. Moreover, it was possible to find lineage-specific MGEs from the same niche, showing acquisition probably due to an evolutionary convergence event or to a genetic group with infective capacity. Furthermore, diversification selection analysis exhibits proteins exposed on the surface subject to selection pressures with an interstrain heterogeneity related to their ability to adapt. This article is the first review describing the genomic relationship to elucidate the diversity and evolution of P. multocida.

Pan-genomic approach shows insight of genetic divergence and pathogenic-adaptation of Pasteurella multocida.

Pasteurella multocida is a gram-negative, non-motile bacterial pathogen, which is associated with chronic and acute infections as snuffles, pneumonia, atrophic rhinitis, fowl cholera and hemorrhagic septicemia. These diseases affect a wide range of domestic animals, leading to significant morbidity and mortality and causing significant economic losses worldwide. Due to the interest in deciphering the genetic diversity and process adaptive between P. multocida strains, this work aimed was to perform a pan-genome analysis to evidence horizontal gene transfer and positive selection among 23 P. multocida strains isolated from distinct diseases and hosts. The results revealed an open pan-genome containing 3585 genes and an accessory genome presenting 1200 genes. The phylogenomic analysis based on the presence/absence of genes and islands exhibit high levels of plasticity, which reflects a high intraspecific diversity and a possible adaptive mechanism responsible for the specific disease manifestation between the established groups (pneumonia, fowl cholera, hemorrhagic septicemia and snuffles). Additionally, we identified differences in accessory genes among groups, which are involved in sugar metabolism and transport systems, virulence-related genes and a high concentration of hypothetical proteins. However, there was no specific indispensable functional mechanism to decisively correlate the presence of genes and their adaptation to a specific host/disease. Also, positive selection was found only for two genes from sub-group hemorrhagic septicemia, serotype B. This comprehensive comparative genome analysis will provide new insights of horizontal gene transfers that play an essential role in the diversification and adaptation mechanism into P. multocida species to a specific disease.

Draft Genome Sequences of Enterohemorrhagic and Enteropathogenic Escherichia coli Strains Isolated from Alpacas in Peru.

The draft genome sequences of two strains of Escherichia coli, isolated from alpacas in Peru, are reported here. ECA1 has been determined to be a strain of enterohemorrhagic E. coli and ECB1 a strain of enteropathogenic E. coli These pathogens are responsible for hemolytic-uremic syndrome in humans and diarrhea in different mammals, respectively.

Draft Genome Sequence of a Virulent Strain of Pasteurella Multocida Isolated From Alpaca.

Pasteurella multocida is one of the most frequently isolated bacteria in acute pneumonia cases, being responsible for high mortality rates in Peruvian young alpacas, with consequent social and economic costs. Here we report the genome sequence of P. multocida strain UNMSM, isolated from the lung of an alpaca diagnosed with pneumonia, in Peru. The genome consists of 2,439,814 base pairs assembled into 82 contigs and 2,252 protein encoding genes, revealing the presence of known virulence-associated genes (ompH, ompA, tonB, tbpA, nanA, nanB, nanH, sodA, sodC, plpB and toxA). Further analysis could provide insights about bacterial pathogenesis and control strategies of this disease in Peruvian alpacas.

Detection and genetic characterization of Pasteurella multocida from alpaca (Vicugna pacos) pneumonia cases.

Pasteurella multocida is a common constituent of upper respiratory tract microbiota but is frequently isolated of alpaca lung tissues from pulmonary infections. Despite its importance, very little is known about this bacteria at molecular level. In order to characterize P. multocida isolates, 24 isolates recovered from 46 mortal acute cases in young alpacas with suspected pneumonia were analyzed, using biochemical and molecular tests for capsule and LPS typing, virulence factors detection, and ERIC-PCR genetic diversity analysis. All the P. multocida isolates belonged to the capsular type A, LPS genotype L6 (related to serotypes 10, 11, 12, and 15), and possessed virulence factors gene toxA and tbpA. ERIC-PCR analysis revealed two electrophoretic profiles, and the majority of isolates (23/24) shared the same fingerprint, indicating strong evidence that there was a common source of infection for all the affect animals. This study revealed the detection of P. multocida type A, LPS genotype L6, and toxA+ and tbpA+ from dead young alpacas with pneumonia in Peru.

Salicola marasensis gen. nov., sp. nov., an extremely halophilic bacterium isolated from the Maras solar salterns in Peru.

Six strains of extremely halophilic bacteria were isolated from several crystallizer ponds of the Maras solar salterns in the Peruvian Andes. On the basis of 16S rRNA gene sequence similarity, G+C contents and DNA-DNA hybridization results, the six isolates constituted a genomically homogeneous group affiliated with the Gammaproteobacteria. The closest relatives were members of the halophilic genera Halovibrio and Halospina, which showed 16S rRNA gene sequence similarities below 97 % and whole-genome hybridization levels below 33 % for the type strain, 7Sm5(T). From the genomic and phenotypic properties of the six novel isolates and phylogenetic reconstruction based on 16S rRNA gene sequence analysis, they can be considered to represent a novel genus within the Gammaproteobacteria. On the basis of the taxonomic study, a novel genus, Salicola gen. nov., is proposed containing the single species Salicola marasensis sp. nov., which is the type species. The type strain of Salicola marasensis is 7Sm5(T) (=CECT 7107(T)=CIP 108835(T)).

Microbial diversity in Maras salterns, a hypersaline environment in the Peruvian Andes.

Maras salterns are located 3,380 m above sea level in the Peruvian Andes. These salterns consist of more than 3,000 little ponds which are not interconnected and act as crystallizers where salt precipitates. These ponds are fed by hypersaline spring water rich in sodium and chloride. The microbiota inhabiting these salterns was examined by fluorescence in situ hybridization (FISH), 16S rRNA gene clone library analysis, and cultivation techniques. The total counts per milliliter in the ponds were around 2 x 10(6) to 3 x 10(6) cells/ml, while the spring water contained less than 100 cells/ml and did not yield any detectable FISH signal. The microbiota inhabiting the ponds was dominated (80 to 86% of the total counts) by Archaea, while Bacteria accounted for 10 to 13% of the 4',6'-diamidino-2-phenylindole (DAPI) counts. A total of 239 16S rRNA gene clones were analyzed (132 Archaea clones and 107 Bacteria clones). According to the clone libraries, the archaeal assemblage was dominated by microorganisms related to the cosmopolitan square archaeon "Haloquadra walsbyi," although a substantial number of the sequences in the libraries (31% of the 16S rRNA gene archaeal clones) were related to Halobacterium sp., which is not normally found in clone libraries from solar salterns. All the bacterial clones were closely related to each other and to the gamma-proteobacterium "Pseudomonas halophila" DSM 3050. FISH analysis with a probe specific for this bacterial assemblage revealed that it accounted for 69 to 76% of the total bacterial counts detected with a Bacteria-specific probe. When pond water was used to inoculate solid media containing 25% total salts, both extremely halophilic Archaea and Bacteria were isolated. Archaeal isolates were not related to the isolates in clone libraries, although several bacterial isolates were very closely related to the "P. halophila" cluster found in the libraries. As observed for other hypersaline environments, extremely halophilic bacteria that had ecological relevance seemed to be easier to culture than their archaeal counterparts.