Rusa deer microbiota: the importance of preliminary data analysis for meaningful diversity comparisons.

The microbiome is an important consideration for the conservation of endangered species. Studies provided evidence of the effect of behavior and habitat change on the microbiota of wild animals and reported various inferences. It indicates the complexity of factors influencing microbiota diversity, including incomplete sampling procedures. Data abnormality may arise due to the procedures warranting preliminary analysis, such as rarefaction, before downstream analysis. This present study demonstrated the effect of data rarefaction and aggregation on the comparison of wild rusa deer's gut microbial diversity. Eighty-five feces samples were collected from 11 deer populations inhabiting three national parks in Java and Bali islands. Using the Illumina Nova-Seq platform, fragments of 16s rRNA gene were sequenced, and raw data of 51,389 reads corresponding to 2 domains, 22 phyla, 45 classes, 83 orders, 182 families, and 460 genera of bacteria were obtained. Data rarefaction was applied at two different library sizes (minimum and fixed) and aggregation (11 populations into 3 research sites) to investigate its effect on the microbial diversity comparison. There are significant differences in alpha diversity between populations, but not research sites, at all library sizes of rarefaction. A similar finding is also found in beta diversity. Moreover, data rarefaction and aggregation result in different values of the diversity metrics. This present study shows that statistical analysis remains a substantial concern in microbiome studies applied to conservation biology. It suggests reporting a more detailed data normalization in microbiome studies as an inherent control of suboptimal sampling, particularly when involving feces.

Selection signatures and formation of the Samosir goat breed through the cultures of the Batak Toba Tribe in Samosir Island, Indonesia.

Background and Aim: The Samosir goat has a high cultural value and is a source of germplasm in Indonesia. This study aimed to reveal the history and selection signatures of the Samosir goat. Materials and Methods: A total of 25 goats were divided into seven subpopulations of Indonesian goat breeds. Deoxyribonucleic acid (DNA) from blood samples was isolated with the use of the gSYNC™ DNA Mini Kit (Geneaid, Taipei, Taiwan). Cytb gene amplification was performed by the polymerase chain reaction (PCR) method, and the PCR products were sequenced. A phylogenetic tree was constructed by the neighbor-joining method using MEGA 11 software. A questionnaire was used to collect information related to the history and breeding practices of the Samosir goat on Samosir Island. Results: Samosir goats are divided into four groups based on their coat color: Completely white, white with brown spots, white with black spots, and white with brown and black spots. The body form of the Samosir goat is similar to that of the Kacang goat. The space below a traditional Toba Batak house is used as a goat pen. The genetic difference between the Samosir goat and the Kacang goat based on the Cytb gene was approximately 0.1%. Conclusion: Phylogenetic analysis between Samosir goats and other indigenous Indonesian goats revealed that Samosir goats form a single clade, with a very close genetic distance from other local goats, such as the Kacang goat. The Toba Batak culture on Samosir Island has significantly influenced the selection and formation of the Samosir goat breed.

Global research alliance in infectious disease: a collaborative effort to combat infectious diseases through dissemination of portable sequencing.

OBJECTIVE: To disseminate the portable sequencer MinION in developing countries for the main purpose of battling infectious diseases, we found a consortium called Global Research Alliance in Infectious Diseases (GRAID). By holding and inviting researchers both from developed and developing countries, we aim to train the participants with MinION's operations and foster a collaboration in infectious diseases researches. As a real-life example in which resources are limited, we describe here a result from a training course, a metagenomics analysis from two blood samples collected from a routine cattle surveillance in Kulan Progo District, Yogyakarta Province, Indonesia in 2019. RESULTS: One of the samples was successfully sequenced with enough sequencing yield for further analysis. After depleting the reads mapped to host DNA, the remaining reads were shown to map to Theileria orientalis using BLAST and OneCodex. Although the reads were also mapped to Clostridium botulinum, those were found to be artifacts derived from the cow genome. An effort to construct a consensus sequence was successful using a reference-based approach with Pomoxis. Hence, we concluded that the asymptomatic cow might be infected with T. orientalis and showed the usefulness of sequencing technology, specifically the MinION platform, in a developing country.

Leptospirosis in Ruminants in Yogyakarta, Indonesia: A Serological Survey with Mixed Methods to Identify Risk Factors.

Leptospirosis is a zoonotic disease occurring worldwide with reproductive symptoms and production losses in livestock, while humans can suffer fatal renal failure. In Yogyakarta Special Province, Indonesia, there have been several outbreaks with high case fatality, demonstrating the public health importance, but there is limited understanding of the epidemiology. This study used an EcoHealth approach to ensure transdisciplinarity and community participation. Seroprevalence of Leptospira in animals was studied between October 2011 and May 2013 in 15 villages. Serum samples from 1404 cattle and 60 small ruminants were screened by a Microscopic Agglutination Test (MAT), first in pools, and then the individual positive samples were identified. Focus group discussions including farmers, village officials, and official stakeholders were used to explore knowledge and behavior of zoonotic diseases, particularly leptospirosis. Two small ruminants were seropositive for Leptospira icterohemorrhagiae. From the cattle, 3.7% were seropositive, and the most common serovars were Leptospira hardjo, followed by L. icterohemorrhagiae. Out of all farms, 5.6% had at least one positive cattle. Risk factor analyses showed that the risk of the farm being seropositive increased if the farmer used water from an open source, or if farming was not the main occupation. This study showed the presence of Leptospira spp. in ruminants in Yogyakarta and identified use of open water as a risk factor for the livestock. We also observed that the knowledge related to leptospirosis was low, and risky farm management practices were commonly employed.

Cryptosporidium Infection Increases the Risk for Chronic Diarrhea Among People Living With HIV in Southeast Asia: A Systematic Review and Meta-Analysis.

We conducted a systematic review research and meta-analysis to reveal the relationship between the risk of chronic diarrhea and Cryptosporidium infection in people living with HIV in Southeast Asia. We performed online peer-reviewed literature research from January 2005 to December 2017, which included PubMed, Science Direct, ProQuest, EBSCO, Cochrane, and Web of Science databases. Calculation of size effects in the meta-analysis was performed by STATA 13.0 software to estimate relative risks (RRs) with 95% confidence intervals (CIs) for any associations. Seven cross-sectional research articles were recruited in this study based on the inclusion and exclusion criteria. Our analysis revealed a significant relationship between cryptosporidiosis and the risk of chronic diarrhea in people living with HIV, with RR = 1.325; 95% CI = 1.157 to 1.517; and P < .000. Our results suggested that cryptosporidiosis increases the risk of chronic diarrhea, and low CD4+ lymphocyte cell counts aggravate the degree of diarrhea. Therefore, clinicians should be more aware in treating HIV-positive people, especially those with low CD4+ cell counts, and we suggest that Cryptosporidium laboratory examinations be conducted immediately.

Strengthening multi-sectoral collaboration on critical health issues: One Health Systems Mapping and Analysis Resource Toolkit (OH-SMART) for operationalizing One Health.

Addressing critical global health issues, such as antimicrobial resistance, infectious disease outbreaks, and natural disasters, requires strong coordination and management across sectors. The One Health approach is the integrative effort of multiple sectors working to attain optimal health for people, animals, and the environment, and is increasingly recognized by experts as a means to address complex challenges. However, practical application of the One Health approach has been challenging. The One Health Systems Mapping and Analysis Resource Toolkit (OH-SMART) introduced in this paper was designed using a multistage prototyping process to support systematic improvement in multi-sectoral coordination and collaboration to better address complex health concerns through an operational, stepwise, and practical One Health approach. To date, OH-SMART has been used to strengthen One Health systems in 17 countries and has been deployed to revise emergency response frameworks, improve antimicrobial resistance national action plans and create multi agency infectious disease collaboration protocols. OH-SMART has proven to be user friendly, robust, and capable of fostering multi-sectoral collaboration and complex system-wide problem solving.

Vitamin D, cell death pathways, and tuberculosis.

BACKGROUND: Mycobacterium tuberculosis induces cellular necrosis that could promote spread of infection. The aim of this study is to analyze the effects of Vitamin D3 supplementation to improve the effectiveness of 2nd-line anti-tuberculosis (TB) drug therapy, especially in relation with cell death pathways. METHODS: Mus musculus C3HeB/FeJ was randomly divided into four groups containing eight animals each. The 1st group (G1), consisting of mice that were intratracheally infected with multidrug-resistant strain of M. tuberculosis and sacrificed on 2-week postinfection to confirm successful infection. (G2) was a group of TB mice without therapy. Then, (G3) was a group of mice with the 2nd-line anti-TB therapy. The last group (G4) was a group of mice receiving not only the 2nd-line anti-TB therapy but also daily oral Vitamin D3 supplementation. Immunohistochemistry was used to measure expression of nuclear Vitamin D receptor, apoptosis marker cleaved caspase-3, cathelin-related antimicrobial peptide (CRAMP) and LC3B autophagy markers, necrosis marker RIPK3, and collagenase matrix metalloproteinase-1 (MMP1). The number of bacteria in the lung was calculated by colony forming units. The partial least square structural equation modeling with SmartPLS 3.2.6 software was used to analyze structural models among the variables. RESULTS: Supplementation of Vitamin D3 on the 2nd-line anti-TB therapy increases Vitamin D3 receptor, CRAMP, LC3B, caspase-3 (P = 0.026, P = 0.000, P= 0.001), presses MMP1, and the number of bacteria (P = 0.010 and P= 0.000, respectively). The structural equation modeling analysis shows that increasing autophagy pathways reduces necrosis by lowering MMP1, whereas apoptosis reduces necrosis by decreasing the number of bacteria (each with indirect effects - 0.543 and - 0.544). CONCLUSION: A comprehensive analysis with the partial least square structural equation modeling shows decreasing necrosis requires increasing autophagy and apoptosis.

Piloting the One Health Systems Mapping and Analysis Resource Toolkit in Indonesia.

As a global network, countries are being asked to meet goals set forth in the Global Health Security Agenda (GHSA) for a workforce capable of effective and efficient prevention, detection and response to infectious disease threats. There is great need for a cross-sectoral workforce that can innovate and problem-solve. To achieve GHSA goals, countries need a way to visualize their existing system, identify opportunities for improvement, and achieve improved cross-sectoral interactions. The One Health Systems Mapping and Analysis Resource Toolkit (OH-SMART) was successfully piloted in West Sumatra, Indonesia, and was used to enhance multi-agency collaboration around infectious disease outbreaks and proved to be an adaptable, scalable process requiring minimal resources. The authors present OH-SMART as a potential tool to help countries analyze their existing health system and create relevant action steps to improve cross-sectoral collaborations.

Native early antigen of Epstein-Barr virus, a promising antigen for diagnosis of nasopharyngeal carcinoma.

The Epstein-Barr virus (EBV) early antigen (EA) complex consists of multiple proteins with relevance for diagnosis of acute, chronic and malignant EBV related diseases, including nasopharyngeal carcinoma (NPC). In a recent study, it was found that the molecular diversity of EBV-specific IgG and IgA antibody responses in NPC patients and demonstrated that these reflect independent B-cell triggering leading to distinct EBV antigen-recognition profiles. The fine-specificity of NPC-related IgG and IgA responses was explored further against defined recombinant and synthetic EBV-EA antigens using immunofluorescence, immunoblot and ELISA techniques and determined their diagnostic value in a large panel of sera from NPC (n = 154), non-NPC tumor patients (n = 133), acute mononucleosis patients (n = 70) and healthy EBV carriers (n = 259). Individual recombinant EBV-EA markers yielded sensitivity/specificity values not exceeding 86%, whereas selected EA-specific peptide epitopes were rather poorly recognized by IgG and IgA antibodies in NPC sera. Surprisingly, we found that a "low salt" native EA-protein extract reproducibly prepared from purified nuclei of EA-induced HH514 cells, and containing characteristic EA(D)-polypeptides, such as p47-54 (BMRF1), p138 (BALF2), p55-DNAse (BGLF5), and p65-TK (BXLF1), but without viral capsid (VCA) or nuclear antigen (EBNA) reactivity, gave highest sensitivity (90.4%) and specificity (95.5%) values for NPC diagnosis in both IgG and IgA ELISA. The data support further the notion that EBV-EA reactive IgG and IgA antibodies in NPC patients are directed against distinct conformational and-in part-linear epitopes on EBV-specific proteins, barely recognized in other EBV-related syndromes. The use of a defined native EBV EA-specific antigen opens the way to further improve serological diagnosis of NPC.