Continuing Orthohantavirus Circulation in Deer Mice in Western Montana

Hantavirus pulmonary syndrome (HPS) is an often-fatal disease caused by New World hantaviruses, such as Sin Nombre orthohantavirus (SNV). In the US, >800 cases of HPS have been confirmed since it was first discovered in 1993, of which 43 were reported from the state of Montana. The primary cause of HPS in the US is SNV, which is primarily found in the reservoir host Peromyscus maniculatus (deer mouse). The reservoir host covers most of the US, including Montana, where multiple studies found SNV in local deer mouse populations. This study aimed to check the prevalence of SNV in the deer mice at popular recreation sites throughout the Bitterroot Valley in Western Montana as compared to previous studies in western Montana. We found high prevalence (up to 20%) of deer mice positive for SNV RNA in the lungs. We were unable to obtain a SNV tissue culture isolate from the lungs but could passage SNV from lung tissue into naïve deer mice. Our findings demonstrate continuing circulation of SNV in western Montana.

Screening of Bacteriophage Encoded Toxic Proteins with a Next Generation Sequencing-Based Assay

Bacteriophage vB_EcoM_fHy-Eco03 (fHy-Eco03 for short) was isolated from a sewage sample based on its ability to infect an Escherichia coli clinical blood culture isolate. Altogether, 32 genes encoding hypothetical proteins of unknown function (HPUFs) were identified from the genomic sequence of fHy-Eco03. The HPUFs were screened for toxic properties (toxHPUFs) with a novel, Next Generation Sequencing (NGS)-based approach. This approach identifies toxHPUF-encoding genes through comparison of gene-specific read coverages in DNA from pooled ligation mixtures before electroporation and pooled transformants after electroporation. The performance and reliability of the NGS screening assay was compared with a plating efficiency-based method, and both methods identified the fHy-Eco03 gene g05 product as toxic. While the outcomes of the two screenings were highly similar, the NGS screening assay outperformed the plating efficiency assay in both reliability and efficiency. The NGS screening assay can be used as a high throughput method in the search for new phage-inspired antimicrobial molecules.

Staphylococcus epidermidis Biofilms Have a High Tolerance to Antibiotics in Periprosthetic Joint Infection

Both Staphylococcus aureus and Staphylococcus epidermidis are commonly associated with periprosthetic joint infections (PJIs). The treatment of PJI can be challenging because biofilms are assumed to have an increased intolerance to antibiotics. This makes the treatment of PJI challenging from a clinical perspective. Although S. aureus has been previously demonstrated to have increased biofilm antibiotic tolerance, this has not been well established with Staphylococcus epidermidis. A prospective registry of PJI S. epidermidis isolates was developed. The efficacy of clinically relevant antibiotics was quantified against these isolates. S. epidermidis planktonic minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were collected using clinical laboratory standard index (CLSI) assays for eight antibiotics (doxycycline, vancomycin, daptomycin, clindamycin, rifampin, nafcillin, and trimethoprim/sulfamethoxazole). Mature biofilms were grown in vitro, after which minimum biofilm inhibitory concentration (MBIC) and minimum biofilm bactericidal concentration (MBBC) were quantified. Only rifampin and doxycycline had a measurable MBIC across all tested isolates. Based on MBBC, 64% of S. epidermidis biofilms could be eliminated by rifampin, whereas only 18% by doxycycline. S. epidermidis biofilm was observed to have a high tolerance to antibiotics as compared to planktonic culture. Isolate biofilm antibiotic tolerance varied to a larger degree than was seen in planktonic cultures.

Nasopharyngeal colonization at birth and the development of early-onset neonatal sepsis

Background Neonatal sepsis is one of the major causes of morbidity and mortality in neonates. Exposure to maternal bacteria during pregnancy or delivery allows for colonization of the normal upper airway. Such bacteria become the major ecological species in the infant. If the colonizing bacteria invade the bloodstream, early-onset neonatal sepsis (EONS) could occur. Objective To evaluate for an association between colonization of the newborn nasopharynx and EONS, as well as for agreement between nasopharyngeal swab culture and blood culture isolate results. Methods This prospective cohort study was conducted in Dr. Wahidin Sudirohusodo General Hospital and Ibnu Sina Hospital, Makassar, South Sulawesi. Nasopharyngeal swab culture was taken within 2 hours of life from newborns who met the inclusion criteria, then they were followed up for signs of EONS. Blood culture was taken from subject with EONS. Results Of the 100 newborns, 69 (69%) had nasopharyngeal bacterial colonization, of whom 5.8% (4/69) experienced EONS. Of the remaining 31 (31%) without colonization, 9.7% (3/31) experienced EONS. There was no significant difference in frequency of EONS between newborns with and without nasopharyngeal colonization. Although Gram-negative bacteria were predominant among colonized newborns, there was no significant difference to numbers of Gram-positive bacteria as a causative agent of EONS. Only one patient with EONS had the same bacterial species in both the nasopharynx and blood culture isolate. Conclusion Newborn nasopharyngeal colonization at birth is not associated with EONS.

Next-Gen sequencing of novel pandemic swine flu [A(H1N1)pdm09] virus in India revealed novel mutations across the genome

The Influenza A H1N1 virus of 2009 was the first pandemic flu virus of the 21st century. Identifying the emergence of mutations in rapidly mutating Influenza viruses that allow increased transmission or confer resistance are invaluable to global outbreak response. Here we recovered 5 complete Influenza A genomes from 4 oropharygeal swabs and one cell culture isolate from a severe Indian outbreak of flu in early 2015. Multiple amino acids substitutions including those known to confer resistance to Oseltamivir and increased pathogenecity in mice were found in the Neuraminidase gene. Additional mutations both reported and novel were found throughout the genome compared to the vaccine strain (California/04/2009). All eight segments of the complete genomes were found to be genetically related to the 2009 pandemic strain, A(H1N1)pdm09 and belonging to the emerging genogroup 6B. This group was found to be of south East Asian origin by time scale phylogentic analysis. A phylogeographic analysis revealed 39 significant migration events among globally circulating viruses. This study is the first extensive complete genome and phylogeographic analysis of 2015 Indian A(H1N1) pdm09 viruses. We report several novel mutations in the 2015 Indian strains which need to be evaluated for effect on viral replication, transmission and resistance to therapy. The identification of mutant A(H1N1)pdm09 from India warrants continuous monitoring of viral evolution for implementation of suitable medical countermeasures.

Determination of Biofilm Formation between Different Strains of Propionibacterium acnes Using Biofilm Assay

Propionibacterium acnes (P. acnes) a member of the normal flora of the skin has constantly been associated with deep tissue infections especially during medical processes. P. acnes have been isolated in deep tissues and are believed to be an aetiological agent in these infections, contributing to the progression of some of these diseases. The biofilm formation ability between different strains of P. acnes was determined. Ten (10) P. acnes clinical isolates were considered, two (2) from acne vulgaris and eight (8) [two (2) per recA types 1A1, 1B, II and III] from lumber herniation tissues. Semi quantitative biofilm analysis using the microtiter plate assay was used with some modification. The semi quantitative biofilm assay was done in triplicates. The result obtained from the biofilm triplicates from 4 days’ incubation using 3 days’ culture showed that isolates 17(IB), 82(IB) and 55(II) showed very high biofilm production for 2 replicates which implies that they are real biofilm producing isolates. Using overnight cultures, higher biofilm production was witnessed with isolates 1(III), Lesion 7 and 84 (IA1) being the highest biofilm producers. Although with 3 days’ culture, isolate 1(III) could easily be discarded as a-non biofilm producer, while lesion 7 and 84 (IA) has been associated to biofilm formation in 3 days’ culture. The production of biofilms by isolates supports the theory that the ability of P. acnes to form biofilms enables it to attach to medical implants hence causing deep tissue infections.

“EFFECT OF ASPERGILLUS SPECIES ON SKIN INFECTION”

We present such a case, manifested by ulceration on skin due to A. niger, which remained undiagnosed for a prolonged period. Although the patient had associated severe fungal infection. Recurrence of the lesion occurred despite repeated anti-fungal therapies. Anti fungal testing was done based on the broth dilution (M-38A, AIIMS, INDIA) method. The culture isolate was found to be sensitive to fluconazole and amphotericin B. Continuation of antifungal therapy improved the symptoms, reducing the size of the lesion. Key words: Aspergillus niger, Skin infection, ulcer, Aspergillus lentulus., Broth dilution.

Complete Genome Sequence of Porcine respirovirus 1 Strain USA/MN25890NS/2016, Isolated in the United States

ABSTRACT A porcine parainfluenza virus type 1 (species Porcine respirovirus 1) cell culture isolate, USA/MN25890NS/2016, was obtained from porcine nasal swabs, and its complete genome sequence (GenBank accession number MF681710) was determined to help further characterize this virus.

Molecular Characterization of a Catalase-Negative Staphylococcus aureus Blood Culture Isolate: TABLE 1

Here we report a catalase-negative methicillin-sensitiveStaphylococcus aureusisolate collected from a blood culture. Sequencing through the gene encoding catalase,katA, demonstrated a 2-bp insertion. The resulting frameshift mutation generates a protein that has lost 26 amino acids (aa) at its C-terminal domain.