Antiviral activity of maca (Lepidium meyenii) against human influenza virus

Objective: To investigate antiviral activity of maca to reduce viral load in kidney (MDCK) cells infected with influenza type A and B viruses (Flu-A and MFalud-inB-, Dreasrpbeyc ctiavneilny)e. Methods: Maca were extracted with methanol (1:2, v/v). The cell viability and toxicity of the eaxgtariancstts Fwluer-eA e avnaldu aFtleud- oBn v MirDusCeKs cwealsls a usssianyge dm uetshinogd aM TteTs ta sfosar yd. eAtenrtmiviinrainl ga ctthiev itiyn hoifb citoimonp oouf nthdes cytopathic effect on cell culture and multiplex RT-PCR. Results: The methanol extract of maca showed low cytotoxicity and inhibited influenza-induced cytopathic effect significantly, while viral load was reduced via inhibition of viral growth in MDCK infected cells. Maca contains potent inhibitors of Flu-A and Flu-B with a selectivity index [cytotoxic concentration 50%/IC50] of 157.4 and 110.5, respectively. Conclusions: In vitro assays demonstrated that maca has antiviral activity not only against Flu-A (like most antiviral agents) but also Flu-B viruses, providing remarkable therapeutic benefits.Financial support of this study was provided by AECID grants (PCI: C/033641/10) and AGAUR (MAT2009-11503, MAT2012-36205, 2009SGR-1208). JDVM support was provided by 1st Concurso Incentivo a la Investigación de la Universidad Peruana de Ciencias Aplicadas, Lima, Peru.Revisión por pare

Carrion's Disease: More Than a Sand Fly-Vectored Illness

Carrion’s disease is a biphasic illness (S1 Fig) caused by an infection of Bartonella bacilliformis, a bacterium that is transmitted through bites of certain phlebotomine sand flies in the Andean valleys of Peru and in some areas of Ecuador and southern Colombia [1,2]. The acute phase, called Oroya fever, is a serious, life-threating illness that mainly affects immunologically naïve populations, such as children. It is also of special concern in pregnant women, because high mortality rates have been described as well as miscarriages, preterm births, and fetal deaths [3]. In this acute phase, the absence or delay of antibiotic treatment may lead to fatal outcomes. In fact, it is considered that, in the pre-antibiotic era, the lethality of this illness ranked between 40% and 88% [1,2]. In the chronic phase, classically considered to occur in previously exposed inhabitants, B. bacilliformis induce endothelial cell proliferation, producing skin lesions called Peruvian warts. In this phase, the lethality is very low [1]. Additionally, the presence of asymptomatic carriers is frequent, although the real numbers remain uncertain because of the difficulty in detecting these subjects

Multi-Locus Sequence Typing of Bartonella bacilliformis DNA Performed Directly from Blood of Patients with Oroya's Fever During a Peruvian Outbreak.

Background Bartonella bacilliformis is the etiological agent of Carrion’s disease, a neglected tropical poverty-linked illness. This infection is endemic of Andean regions and it is estimated that approximately 1.7 million of South Americans are at risk. This bacterium is a fastidious slow growing microorganism, which is difficult and cumbersome to isolate from clinical sources, thereby hindering the availability of phylogenetic relationship of clinical samples. The aim of this study was to perform Multi Locus Sequence Typing of B. bacilliformis directly in blood from patients diagnosed with Oroya fever during an outbreak in Northern Peru. Methodology/Principal Findings DNA extracted among blood samples from patients diagnosed with Oroya’s fever were analyzed with MLST, with the amplification of 7 genetic loci (ftsZ, flaA, ribC, rnpB, rpoB, bvrR and groEL) and a phylogenetic analysis of the different Sequence Types (ST) was performed. A total of 4 different ST were identified. The most frequently found was ST1 present in 66% of samples. Additionally, two samples presented a new allelic profile, belonging to new STs (ST 9 and ST 10), which were closely related to ST1. Conclusions/Significance The present data demonstrate that B. bacilliformis MLST studies may be possible directly from blood samples, being a promising approach for epidemiological studies. During the outbreak the STs of B. bacilliformis were found to be heterogeneous, albeit closely related, probably reflecting the evolution from a common ancestor colonizing the area. Additional studies including new samples and areas are needed, in order to obtain better knowledge of phylogenetic scenario B. bacilliformisThis work has been supported by the Spanish Network for the Research in Infectious Diseases [REIPI RD12/0015],by Generalitat de Catalunya, Departament d’Universitats, Recerca i Societat de la Informació [2014 SGR 26] and by by a grant of the Instituto de Salud Carlos III - Spain (PI11/ 00983) which included FEDER funds (JR). MJP has a postdoctoral fellowship from CONCYTEC/ FONDECYT (grant number: CG05-2013- FONDECYT). JR has a fellowship from the programRevisión por pare

Detection of human Metapneumovirus infection in children under 18 years old hospitalized in Lima-Peru

Human Metapneumovirus (hMPV) is a negative single-stranded RNA virus. Infection by hMPV mainly affects the pediatric population and can cause upper or lower respiratory tract pathologies which can develop life threating complications. This study was carried out between 2009 and 2010 in a high complexity national hospital in Lima, Peru. The time frame corresponds to the pandemic of influenza A H1N1. Methods. A prospective study was performed between September 2009 and September 2010. Patients with a clinical diagnosis suggestive of an acute respiratory infection were included. RT-PCR was utilized to attain the amplification and identification of the hMPV. Results.A total of 539 samples were analyzed from patients with a clinical context suggestive of an acute respiratory tract infection. Of these samples 73, (13.54%) were positive for hMPV. Out of the positive cases, 63% were under one year old, and increased to nearly 80% when considering children younger than two years old. Cough was the most frequent symptom presented by our population with a number of 62 cases (84.93%). Viral seasonality was also established, noting its predominance during the months of summer in the southern hemisphere. The infection by hMPV has an important prevalence in Peru. It mainly affects children under one year old and should be considered an important differential diagnosis in a patient with an acute respiratory infection.Postprint (author's final draft

Sera of Peruvians with fever of unknown origins include viral nucleic acids from non-vertebrate hosts

Serum samples collected from 88 Peruvians with unexplained fever were analyzed for viral sequences using metagenomics. Nucleic acids of anelloviruses, pegivirus A (GBV-C), HIV, Dengue virus, and Oropouche virus were detected. We also characterized from two sera the RNA genomes of new species of partitivirus and dicistrovirus belonging to viral families known to infect fungi or arthropod, respectively. Genomic DNA of a putative fungal cellular host could be PCR amplified from the partitivirus-containing serum sample. The detection in human serum of nucleic acids from viral families not known to infect vertebrates may indicate contamination during sample collection and aliquoting or human infection by their presumed cellular host, here a fungus. The role, if any, of the non-vertebrate infecting viruses detected in serum in inducing fever is unknown.Peer reviewe

Unlocking SARS-CoV-2 detection in low- and middle-income countries

Low- and middle-income countries (LMICs) are significantly affected by SARS-CoV-2, partially due to their limited capacity for local production and implementation of molecular testing. Here, we provide detailed methods and validation of a molecular toolkit that can be readily produced and deployed using laboratory equipment available in LMICs. Our results show that lab-scale production of enzymes and nucleic acids can supply over 50,000 tests per production batch. The optimized one-step RT-PCR coupled to CRISPR-Cas12a-mediated detection showed a limit of detection of 102 ge/μL in a turnaround time of 2 h. The clinical validation indicated an overall sensitivity of 80%–88%, while for middle and high viral load samples (Cq ≤ 31) the sensitivity was 92%–100%. The specificity was 96%–100% regardless of viral load. Furthermore, we show that the toolkit can be used with the mobile laboratory Bento Lab, potentially enabling LMICs to implement detection services in unattended remote regions.Fondo Nacional de Desarrollo Científico, Tecnológico y de Innovación TecnológicaRevisión por pare

Development and characterisation of highly antibiotic resistant Bartonella bacilliformis mutants

The objective was to develop and characterise in vitro Bartonella bacilliformis antibiotic resistant mutants. Three B. bacilliformis strains were plated 35 or 40 times with azithromycin, chloramphenicol, ciprofloxacin or rifampicin discs. Resistance-stability was assessed performing 5 serial passages without antibiotic pressure. MICs were determined with/without Phe-Arg-beta-Napthylamide and artesunate. Target alterations were screened in the 23S rRNA, rplD, rplV, gyrA, gyrB, parC, parE and rpoB genes. Chloramphenicol and ciprofloxacin resistance were the most difficult and easiest (>37.3 and 10.6 passages) to be selected, respectively. All mutants but one selected with chloramphenicol achieved high resistance levels. All rifampicin, one azithromycin and one ciprofloxacin mutants did not totally revert when cultured without antibiotic pressure. Azithromycin resistance was related to L4 substitutions Gln-66 --> Lys or Gly-70 --> Arg; L4 deletion Delta62-65 (Lys-Met-Tyr-Lys) or L22 insertion 83::Val-Ser-Glu-Ala-His-Val-Gly-Lys-Ser; in two chloramphenicol-resistant mutants the 23S rRNA mutation G2372A was detected. GyrA Ala-91 --> Val and Asp-95 --> Gly and GyrB Glu474 --> Lys were detected in ciprofloxacin-resistant mutants. RpoB substitutions Gln-527 --> Arg, His-540 --> Tyr and Ser-545 --> Phe plus Ser-588 --> Tyr were detected in rifampicin-resistant mutants. In 5 mutants the effect of efflux pumps on resistance was observed. Antibiotic resistance was mainly related to target mutations and overexpression of efflux pumps, which might underlie microbiological failures during treatments

Small Circular Rep-Encoding Single-Stranded DNA Genomes in Peruvian Diarrhea Virome

Metagenomic analysis of diarrhea samples revealed the presence of numerous human enteric viruses and small circular Rep-encoding single-stranded DNA (CRESS-DNA) genomes. One such genome was related to smacoviruses, while eight others were related to genomes reported in the feces of different mammals. The tropism of these CRESS-DNA viruses remains unknown.Revisión por pare

Identification of human papillomavirus as a preventive strategy for cervical cancer in asymptomatic women in the Peruvian Andes

Objective: To detect the most prevalent human papillomavirus (HPV) genotypes samples of asymptomatic Peruvian women by analyzing the correlation betwe ienn c Pearvpiacnailc somlaeoaur (PAP)-stained cervical tests and PCR-sequencing. Methods: A total of 254 women attending routine gynecological examinations were included in pthaitsh ostluogdiys.t Tahned scalmaspsliefise dw ebrye tahnea Blyeztehde sbdya PsAysPt etmec.h HnPiqVu ea manpdli feicxaatmioinn ewda su nddoenre au msinicgr othsceo pprei mbeyr as specific for E1 region and positive specimens were confirmed by direct sequencing. Results: The prevalence of HPV was investigated in 254 cervical scrape samples by PCR. PAP smear showed that 94.9% cases had normal morphology and 5.1% had an inflammatory pattern; 2p0r.e5v%a lwenert eg efonuontydp teo ibne c ionrfreeclatetido nw iwthit hH PchVa, ncgoems pinri scienrgv i2c0a dl icfyfetorelongt yg.enotypes. HPV16 was the most Conclusions: Our results suggest the HPV is very frequent even in women with negative PAP, eannddo PceCrRvi csaele smasm tpol ebs.e Itdheen tbifeicsat toiopnt ioofn t htoe HdePtVe rgmeinnoety tphee inc aaussyamtipvteo magateinc tw oofm HePnV m ianyf eaclltoiown t hine nimatpulreaml henisttaotriyo no fo tfh ea pdpisroepasreia aten dp rthope hsyulbascetqicu emnet adseuvreelso pwmheicnht omf acye rhviacvael ma adliirgencatn [email protected] work has been partially supported by Universidad Peruana de Ciencias Aplicadas (UPC), Instituto de Investigación Nutricional and Instituto de Investigación de Efrnomfe rtmheed pardoegsr aImnf ecciosas, Lima, Peru. JR has a fellowship I3, of the ISCIII (Grant No. CES11/012), and LJDV from the Generalitat de Catalunya (2009SGR1208).Revisión por pare

Antibacterial activity of Myrciaria dubia (Camu camu) against Streptococcus mutans and Streptococcus sanguinis

AbstractObjectiveTo evaluate the antibacterial and cytotoxic effect of Myrciaria dubia (Camu camu) (M. dubia) methanol extract, against Streptococcus mutans (ATCC 25175) (S. mutans) and Streptococcus sanguinis (ATCC 10556) (S. sanguinis).MethodsTwo methanol extracts of M. dubia were prepared in vitro, from the seeds and pulp. Ten independent tests were prepared for each type of extract, using 0.12% chlorhexidine solution as positive control. Agar diffusion test was used by preparing wells with the experimental solutions cultivated in anaerobic conditions for 48 h at 37 °C. Meanwhile, the minimum inhibitory concentration and the cytotoxic effect over MDCK cell line was found.ResultsA higher antibacterial effect was observed with the methanol seed extract with an inhibitory halo of (21.36 ± 6.35) mm and (19.21 ± 5.18) mm against S. mutans and S. sanguinis, respectively. The methanol extract of the pulp had an effect of (16.20 ± 2.08) mm and (19.34 ± 2.90) mm, respectively. The minimum inhibitory concentration of the pulp extract was 62.5 μg/mL for both strains, whereas for the seed antibacterial activity was observed even at low concentrations. The CC50 of the seeds extract was at a higher concentration than 800 μg/mL and 524.37 μg/mL for the pulp extract.ConclusionsThe experimental findings demonstrated the antibacterial effect of the methanol extract of M. dubia against S. mutans and S. sanguinis. These extracts were not cytotoxic at high concentrations