Differentially represented proteins in response to infection with Mycobacterium tuberculosis identified by quantitative serum proteomics in Asian elephants

Tuberculosis is a major global concern. Tuberculosis in wildlife is a risk for zoonotic transmission and becoming one of the challenges for conservation globally. In elephants, the number of cases is likely rising. The aim of this study was to identify proteins related to tuberculosis infection in elephants, which could then be used for the development of diagnostic tools and/or vaccines. A serum proteomics approach was used to characterize differentially represented proteins in response to Mycobacterium tuberculosis in Asian elephants (Elaphas maximus). Blood samples were collected from eight elephants, four of which were antibody positive for tuberculosis and four were antibody negative. Proteomics analysis identified 26 significantly dysregulated proteins in response to tuberculosis. Of these, 10 (38%) were identified as immunoglobulin and 16 (62%) as non-immunoglobulin proteins. The results provided new information on the antibody response to mycobacterial infection and biomarkers associated with tuberculosis and protective response to mycobacteria in Asian elephants. Protective mechanisms included defense against infection (Alpha- 1-B glycoprotein A1BG, Serpin family A member 1 SERPINA1, Transthyretin TTR), neuroprotection (TTR), and reduced risks of inflammation, infections, and cancer (SERPINA1, Keratin 10 KRT10). Using a translational biotechnology approach, the results provided information for the identification of candidate diagnostic, prognostic, and protective antigens for monitoring and control of tuberculosis in Asian elephants.La tuberculosis es una de las principales preocupaciones a nivel mundial. La tuberculosis en la fauna salvaje es un riesgo de transmisión zoonótica y se está convirtiendo en uno de los retos de la conservación a nivel mundial. En los elefantes, es probable que el número de casos aumente. El objetivo de este estudio era identificar las proteínas relacionadas con la infección por tuberculosis en los elefantes, que podrían utilizarse para el desarrollo de herramientas de diagnóstico y/o vacunas. Se utilizó un enfoque de proteómica sérica para caracterizar las proteínas representadas diferencialmente en respuesta a Mycobacterium tuberculosis en elefantes asiáticos (Elaphas maximus). Se recogieron muestras de sangre de ocho elefantes, cuatro de los cuales dieron positivo a los anticuerpos de la tuberculosis y cuatro fueron negativos a los anticuerpos. El análisis proteómico identificó 26 proteínas significativamente desreguladas en respuesta a la tuberculosis. De ellas, 10 (38%) se identificaron como inmunoglobulinas y 16 (62%) como proteínas no inmunoglobulínicas. Los resultados aportaron nueva información sobre la respuesta de los anticuerpos a la infección micobacteriana y los biomarcadores asociados a la tuberculosis y la respuesta protectora a las micobacterias en los elefantes asiáticos. Los mecanismos de protección incluían la defensa contra la infección (glicoproteína alfa-1-B, miembro 1 de la familia A de las serpinas SERPINA1, transtiretina TTR), la neuroprotección (TTR) y la reducción del riesgo de inflamación, infecciones y cáncer (SERPINA1, queratina 10 KRT10). Utilizando un enfoque de biotecnología traslacional, los resultados proporcionaron información para la identificación de antígenos candidatos de diagnóstico, pronóstico y protección para el seguimiento y control de la tuberculosis en elefantes asiáticos

Supplementary material for Phylogenomic analysis supports Mycobacterium tuberculosis transmission between humans and elephants

Supplementary information: Table S1: Detailed information on the 94 whole genomes retrieved from GenBank and used in this study.Peer reviewe

Phylogenomic analysis supports Mycobacterium tuberculosis transmission between humans and elephants

[Introduction]: Tuberculosis is an infectious disease caused by a group of acid-fast bacilli known as Mycobacterium tuberculosis complex (MTC), which has a major impact on humans. Transmission of MTC across the human-animal interface has been demonstrated by several studies. However, the reverse zoonotic transmission from humans to animals (zooanthroponosis) has often been neglected.[Methods]: In this study, we used Nanopore MinION and Illumina MiSeq approaches to sequence the whole genome of M. tuberculosis strains isolated from two deceased Asian elephants (Elephas maximus) and one human in Chitwan, Nepal. The evolutionary relationships and drug resistance capacity of these strains were assessed using the whole genome data generated by the stand-alone tool Tb-Profiler. Phylogenomic trees were also constructed using a non-synonymous SNP alignment of 2,596 bp, including 94 whole genome sequences representative of the previously described M. tuberculosis lineages from elephants worldwide (lineages 1 and 4) and from humans in Nepal (lineages 1, 2 and 3).[Results and discussion]: The new genomes achieved an average coverage of 99.6%, with an average depth of 55.67x. These M. tuberculosis strains belong to lineage 1 (elephant DG), lineage 2 (elephant PK) and lineage 4 (human), and none of them were found to have drug-resistant variants. The elephant-derived isolates were evolutionarily closely related to human-derived isolates previously described in Nepal, both in lineages 1 and 2, providing additional support for zooanthroponosis or bidirectional transmission between humans and elephants. The human-derived isolate clustered together with other published human isolates from Argentina, Russia and the United Kingdom in the lineage 4 clade. This complex multi-pathogen, multi-host system is challenging and highlights the need for a One Health approach to tuberculosis prevention and control at human-animal interface, particularly in regions where human tuberculosis is highly endemic.This work was supported by National Funds through FCT-Fundação para a Ciência e a Tecnologia in the scope of the project UIDP/50027/2020.Peer reviewe

Natural Clerodendrum-derived tick repellent: learning from Nepali culture

Ticks attaching to ear canals of humans and animals are the cause of otoacariasis, common in rural areas of Nepal. The plant Clerodendrum viscosum is used in multiple indigenous systems of medicine by ethnic communities in the Indo-Nepali-Malaysian region. Visiting the Chitwan National Park, we learned that in indigenous medicine, flower extract of C. viscosum is utilized to treat digestive disorders and extracts from leaves as tick repellent to prevent ticks from invading or to remove them from the ear canal. The objective of our study was to provide support to indigenous medicine by characterizing the in vivo effect of leave extracts on ticks under laboratory conditions and its phytochemical composition. We collected plant parts of C. viscosum (leaves and flowers) and mango (Mangifera indica) leaves at the Chitwan National Park, previously associated with repellent activity to characterize their effect on Ixodes ricinus ticks by in vivo bioassays. A Q-ToF high-resolution analysis (HPLC-ESI-QToF) was conducted to elucidate phenolic compounds with potential repellent activity. Clerodendrum viscosum and M. indica leaf extracts had the highest tick repellent efficacy (%E = 80–100%) with significant differences when compared to C. viscosum flowers extracts (%E = 20–60%) and phosphate-buffered saline. Phytochemicals with tick repellent function as caffeic acid, fumaric acid and p-coumaric acid glucoside were identified in C. viscosum leaf extracts by HPLC-ESI-QToF, but not in non-repellent flower extracts. These results support the Nepali indigenous medicine application of C. viscosum leaf extracts to repel ticks. Additional research is needed for the development of natural and green repellent formulations to reduce the risks associated with ticks resistant to acaricides.Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature.Peer reviewe

Prevalence of viral load suppression and acquired drug resistance among people living with HIV in Nepal: a nationally representative surveillance study

ABSTRACT: Objectives: This is the first nationally representative study to estimate the prevalence of viral load (VL) suppression and acquired HIV drug resistance (ADR) among people living with HIV (PLHIV) in Nepal. Methods: A cross-sectional study recruited 1418 PLHIV from 20 ART centres in Nepal, using a two-stage cluster design. Participants were eligible if they were HIV-positive individuals on ART for 9–15 months or at least 48 months. Plasma specimens were collected and tested for the quantification of HIV-1 RNA. Specimens with a VL ≥1000 copies/mL were further processed for sequencing of PR and RT genes of HIV-1. The sequences were then analysed to detect mutations causing HIV drug resistance. Results: The prevalence of ADR was 3.7% (95% confidence interval [CI]: 1.8–7.6) and 3.0% (95% CI: 1.8–5.2) among PLHIV who received ART for 9–15 months and 48 months or more, respectively. The prevalence of VL suppression was 95.3% (95% CI: 91.7–97.4) among those on ART for 9–15 months, and 96.5% (95% CI: 94.7–97.7) among those on ART for at least 48 months. The prevalence of any detectable acquired resistance to antiretroviral drugs was 80.7% (95% CI: 58.6–92.5) among those on ART for 9–15 months with VL ≥1000 copies/mL and 81.6% (95% CI: 55.4–94.0) among those on ART for at least 48 months with VL ≥1000 copies/mL. Conclusion: This study suggests that improved accessibility to VL monitoring and timely assessment of drug resistance in routine HIV programs are crucial in Nepal to ensure access to HIV treatment for all in need

Understanding Mycobacterium tuberculosis complex in elephants through a One Health approach: a systematic review

[Background]: Mycobacterium tuberculosis complex (MTC) that causes the chronic infectious disease- tuberculosis (TB), often presents with a complicated epidemiological pattern where the transmission chain may include humans, domestic animals and wildlife, including elephants. TB has been reported globally in both captive and wild elephants. The One Health approach might be the most effective way of understanding the shared MTC infection dynamics in captive and wild animals like Asian elephants. This systematic review accumulates evidence on occurrence, transmission pathways, and preventive measures of TB in elephants from a One Health perspective.[Results]: The prevalence of TB reported in elephant populations ranges from 0 to 23.33% and high prevalence's are reported for elephants that are in close proximity to infected humans. The risk of elephant to human infection transmission increased significantly with exposure duration and contact with infected elephants. Some studies described the plausible TB transmission to captive elephants from other animals (wild and domestic), suggesting inter- and intra-species transmission. The results of this systematic review based on 27 relevant published works, suggest three overarching interrelated transmission pathways for M. tuberculosis infections in Asian elephants- i) humans and elephants, ii) other animals (wild or domestic) and elephants and iii) unclear sources of infection.[Conclusions]: The progress made with new TB diagnostic tools provides multiple methods to choose from. However, lack of harmonization of TB testing in elephants and their human contacts remains a challenge to prevent TB in those animals. Routine TB screening among elephants and caretakers by setting up an occupational health program for early diagnosis of infection through combined efforts of public health, veterinary medicine, and occupational health experts is suggested. This implies the need for a One Health approach to elephant TB control. This review reveals the need for more research on Mycobacterium tuberculosis complex transmission pathways at the human-animal interface.Peer reviewe

Surveillance of Influenza A Virus and Its Subtypes in Migratory Wild Birds of Nepal

Nepal boarders India and China and all three countries lie within the Central Asian Flyway for migratory birds. Novel influenza A H7N9 caused human fatalities in China in 2013. Subclinical infections of influenza A H7N9 in birds and the potential for virus dispersal by migratory birds prompted this study to assess avian H7N9 viral intrusion into Nepal. Surveillance of influenza A virus in migratory birds was implemented in early 2014 with assistance from the Food and Agricultural Organization (FAO). Of 1811 environmental fecal samples collected from seven wetland migratory bird roosting areas, influenza A H9N2 was found in one sample from a ruddy shelduck in Koshi Tappu Wildlife Reserve located in southern Nepal. Avian H7N9 and other highly pathogenic avian influenza viruses were not detected. This study provides baseline data on the status of avian influenza virus in migratory bird populations in Nepal

Collection sites and sample sizes for Influenza A virus surveillance of migratory birds in Nepal.

<p>Collection sites and sample sizes for Influenza A virus surveillance of migratory birds in Nepal.</p

Phylogenetic tree of the HA gene of LPAI H9N2 virus from Nepal.

<p>The tree is generated by neighbor-joining method in MEGA 6. Numbers at the branches indicate bootstrap values; only values > 70 are shown.</p

General flyways used by migratory shorebird species.

<p>Reprinted from Food and Agriculture Animal Production and Health Manual No. 5[<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133035#pone.0133035.ref027" target="_blank">27</a>] under a CC BY license, with permission from FAO copyright unit (2007).</p