Comparing the respiratory tract microbiome in captive elephants and humans in Chitwan National Park: Implications for conservation medicine.

In order to ascertain the comprehensive prevalence of gastrointestinal parasites among captive Asian elephants in Chitwan National Park, a cross-sectional investigation was conducted. A total of 103 samples was purposefully collected. Demographic details encompassing age and gender, along with epidemiological information concerning deworming status, timing intervals, and nutritional condition for both government-owned and privately-owned elephants, were procured through a structured questionnaire survey. The process involved microscopic identification and quantification of gastrointestinal parasites through sedimentation, centrifugal floatation, and MacMaster Egg Per Gram (EPG) count methods. The resultant data indicated an overall prevalence of gastrointestinal parasites at 47.57% (49 out of 103 samples). The dominant class of parasites observed was Nematodes (n=30, 61.22%), followed by Trematodes (n=14, 28.57%) and Cestodes (n=5, 10.20%). Six distinct parasite genera were identified with positive results: Strongylus (26.53%), Trichostrongylus (24.48%), Fasciola (16.35%), Paramphistomum (12.24%), Anoplocephala (10.20%), and Ascaris (10.20%). Notably, the prevalence was markedly higher in females (39.80%) in comparison to males (7.76%), with the disparity being statistically significant (p>0.05). Additionally, a noteworthy correlation was observed between parasite prevalence, age groups, and deworming history, with statistical significance (p<0.05). The Egg Per Gram (EPG) count analysis demonstrated that the majority (87.75%) of the positively identified samples exhibited mild infection (100-500 eggs), while a relatively low percentage (6.12%) displayed heavy infection (1000-1500 eggs). The mean EPG was calculated as (248.39 ± 54.25). Consequently, the heightened prevalence of gastrointestinal parasites in captive elephants within Chitwan National Park underscores the necessity for targeted interventions to mitigate the risk of parasitic infestations.

BackgroundThe gut microbiome is important to immune health, metabolism, and hormone regulation. Understanding host–microbiome relationships in captive animals may lead to mediating long term health issues common in captive animals. For instance, zoo managed African elephants (Loxodonta africana) and Asian elephants (Elephas maximus) experience low reproductive rates, high body condition, and gastrointestinal (GI) issues. We leveraged an extensive collection of fecal samples and health records from the Elephant Welfare Study conducted across North American zoos in 2012 to examine the link between gut microbiota and clinical health issues, reproductive hormones, and metabolic hormones in captive elephants. We quantified gut microbiomes of 69 African and 48 Asian elephants from across 50 zoos using Illumina sequencing of the 16S rRNA bacterial gene.ResultsElephant species differed in microbiome structure, with African elephants having lower bacterial richness and dissimilar bacterial composition from Asian elephants. In both species, bacterial composition was strongly influenced by zoo facility. Bacterial richness was lower in African elephants with recent GI issues, and richness was positively correlated with metabolic hormone total triiodothyronine (total T3) in Asian elephants. We found species-specific associations between gut microbiome composition and hormones: Asian elephant gut microbiome composition was linked to total T3 and free thyroxine (free T4), while fecal glucocorticoid metabolites (FGM) were linked to African elephant gut microbiome composition. We identified many relationships between bacterial relative abundances and hormone concentrations, including Prevotella spp., Treponema spp., and Akkermansia spp.ConclusionsWe present a comprehensive assessment of relationships between the gut microbiome, host species, environment, clinical health issues, and the endocrine system in captive elephants. Our results highlight the combined significance of host species-specific regulation and environmental effects on the gut microbiome between two elephant species and across 50 zoo facilities. We provide evidence of clinical health issues, reproductive hormones, and metabolic hormones associated with the gut microbiome structure of captive elephants. Our findings establish the groundwork for future studies to investigate bacterial function or develop tools (e.g., prebiotics, probiotics, dietary manipulations) suitable for conservation and zoo management.

The gut microbiome affects the physical and mental wellbeing of an animal. Several factors, including diet, host physiology, age, sex, lifestyle, and environmental factors, influence the dynamic gut microbiome. We studied the gut microbiome composition of the endangered Asian elephants (Elephas maximus) kept under prolonged captive conditions (15.8 ± 3.9years) and their wild counterparts, as both were exposed to two different environmental pressures. A total of 648,581 high-quality sequences were obtained, comprising 208 microbial families from 22 phyla and 97 orders, as determined by high-throughput 16S rRNA gene sequencing. Among them, 90% of the microbes belonged to the phyla Firmicutes, Proteobacteria and Bacteroidetes. Our analysis revealed a distinct variation in the gut microbiome between captive and wild elephants. The captive elephants had a higher abundance of the microbial phyla Kiritimatiellaeota, Tenericutes, Euryarchaeota, and Verrucomicrobia, which suggests that captivity alters the gut microbiome. These findings reveal distinct patterns of gut microbiome diversity between captive and wild elephants, underscoring the role of diet and environmental conditions in shaping the elephant gut microbiome.

Abstract. Gautum S, Koju NP. 2022. Demographic and health status of captive elephants around Chitwan National Park, Nepal. Biodiversitas 23: 1621-1627. Elephants have been captive from ancient times. As a keystone species, it requires proper care and management to sustain its viable population in the wild as well as a captive. However, there is a lack of study on captive elephants in Nepal. This study aimed to explore the status of captive elephants in Sauraha, Chitwan National Park, the top tourist destination of Nepal. Field survey, Key-Informant Interview (KII), and questionnaire survey were carried out for the primary source of data collection. During which, 78 mahouts were interviewed. The study revealed 97 (16 males and 81 females) captive elephants in Sauraha during the study period. Among them, 17 were born by captive mothers. The distance traveled by an elephant has recorded 16 km for minimum and 84 km in maximum per day and carries the average weight of 450 kg. Captive elephants belonging to the private company (N=60) were used only in tourism, while captive elephants from National Trust for Nature Conservation (NTNC) and Governments were used in patrolling and transportation. The most common disease in captive elephants is tuberculosis (27%). Similarly, parasitic infections, diseases like foot problems, and colic were also recorded. An effective policy and plan are urgently required to conserve this endangered umbrella species in captivity.

The fatty acid components of the plasma triglycerides and the phospholipid fractions of the red blood cells of a captive group of two African (Loxodonta africana) and four Asian (Elephas...

IntroductionSevere acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has infected a substantial portion of the world’s population, and novel consequences of COVID-19 on the human body are continuously being uncovered. The human microbiome plays an essential role in host health and well-being, and multiple studies targeting specific populations have reported altered microbiomes in patients infected with SARS-CoV-2. Given the global scale and massive incidence of COVID on the global population, determining whether the effects of COVID-19 on the human microbiome are consistent and generalizable across populations is essential.MethodsWe performed a synthesis of human microbiome responses to COVID-19. We collected 16S rRNA gene amplicon sequence data from 11 studies sampling the oral and nasopharyngeal or gut microbiome of COVID-19-infected and uninfected subjects. Our synthesis included 1,159 respiratory (oral and nasopharyngeal) microbiome samples and 267 gut microbiome samples from patients in 11 cities across four countries.ResultsOur reanalyses revealed communitywide alterations in the respiratory and gut microbiomes across human populations. We found significant overall reductions in the gut microbial diversity of COVID-19-infected patients, but not in the respiratory microbiome. Furthermore, we found more consistent community shifts in the gut microbiomes of infected patients than in the respiratory microbiomes, although the microbiomes in both sites exhibited higher host-to-host variation in infected patients. In respiratory microbiomes, COVID-19 infection resulted in an increase in the relative abundance of potentially pathogenic bacteria, including Mycoplasma.DiscussionOur findings shed light on the impact of COVID-19 on the human-associated microbiome across populations, and highlight the need for further research into the relationship between long-term effects of COVID-19 and altered microbiota.

Elephants are endangered species and threatened with extinction. They are monogastric herbivorous, hindgut fermenters and their digestive strategy requires them to consume large amounts of low quality forage. The gut microbiome is important to their metabolism, immune regulation, and ecological adaptation. Our study investigated the structure and function of the gut microbiota as well as the antibiotic resistance genes (ARGs) in captive African and Asian elephants on the same diet. Results showed that captive African and Asian elephants had distinct gut bacterial composition. MetaStats analysis showed that the relative abundance of Spirochaetes (FDR = 0.00) and Verrucomicrobia (FDR = 0.01) at the phylum level as well as Spirochaetaceae (FDR = 0.01) and Akkermansiaceae (FDR = 0.02) at the family level varied between captive African and Asian elephants. Among the top ten functional subcategories at level 2 (57 seed pathway) of Kyoto Encyclopedia of Genes and Genomes (KEGG) database, the relative gene abundance of cellular community-prokaryotes, membrane transport, and carbohydrate metabolism in African elephants were significantly lower than those in Asian elephants (0.98 vs. 1.03%, FDR = 0.04; 1.25 vs. 1.43%, FDR = 0.03; 3.39 vs. 3.63%; FDR = 0.02). Among the top ten functional subcategories at level 2 (CAZy family) of CAZy database, MetaStats analysis showed that African elephants had higher relative gene abundance of Glycoside Hydrolases family 28 (GH 28) compared to Asian elephants (0.10 vs. 0.08%, FDR = 0.03). Regarding the antibiotic resistance genes carried by gut microbes, MetaStats analysis showed that African elephants had significantly higher relative abundance of vanO (FDR = 0.00), tetQ (FDR = 0.04), and efrA (FDR = 0.04) than Asian elephants encoding resistance for glycopeptide, tetracycline, and macrolide/rifamycin/fluoroquinolone antibiotic, respectively. In conclusion, captive African and Asian elephants on the same diet have distinct gut microbial communities. Our findings established the ground work for future research on improving gut health of captive elephants.

Elephant Endotheliotropic Herpesvirus (EEHV) is a highly destructive viral disease affecting both Asian and African elephants, posing a threat in captivity and the wild. Despite habitat loss, poaching, and human-elephant conflict being major concerns, EEHV emerges as a significant threat to elephant species' long-term survival. Until now, no reported cases of EEHV have been documented in Malaysia, particularly in Peninsular Malaysia. This raises questions about whether the absence of report is due to the virus's actual absence or a lack of awareness. Wildlife disease surveillance programmed initiated EEHV screening in Asian elephant at National Elephant Conservation Centre (PKGK) which one of establish ex-situ facilities for elephant conservation in Peninsular Malaysia. Molecular detection approach by utilizing polymerase chain reaction amplification targeting polymerase U38 gene region revealed 12 individuals out of 33 captive Asian elephants was found EEHV-positive, spanning ages from 2 months to 15 years and both sexes. Phylogenetic analysis revealed distinct EEHV subtypes identified in this this study. Despite being EEHV-positive, the captive Asian elephants displayed good health, indicating asymptomatic infections of EEHV1A, EEHV4, and EEHV5. This marks the initial identification of EEHV and subclinical infections in captive elephants in Peninsular Malaysia, emphasizing the need for urgent and enhanced health care management to safeguard these magnificent animals in Peninsular Malaysia for better conservation management strategies in the future

Background and Aim:Brucellosis is considered as an important zoonotic disease caused by various strains of Brucella in numerous host species. Although brucellosis has been reported in almost animal species, the relevance of brucellosis infection and diagnostic technique in Asian elephant (Elephas maximus) has been limited. The present serological investigation aimed to investigate the antibody response to Brucella abortus in captive Asian elephants in North Thailand. Moreover, further serological survey was also conducted to detect the antibody response to Brucella canis in stray dogs cohabiting the same area as the elephant herd.Materials and Methods:Serum samples were collected from 40 captive Asian elephants and submitted for serological analysis based on B. abortus antigen using Rose Bengal plate test (RBPT) in combination with ethylenediaminetetraacetic acid-tube agglutination test (EDTA-TAT) as a supplementary test and by commercial indirect enzyme-linked immunosorbent assay (iELISA). In addition, serum samples were also obtained from 16 stray dogs that live nearby the elephant-raising area and were tested using commercial Dot-ELISA based on B. canis antigen.Results:Serological analysis in captive Asian elephants showed 100% seronegative (40/40) from all serological tests response to B. abortus. For stray dogs, 12.5% (2/16) had a low positive reaction response to B. canis.Conclusion:The serological survey for brucellosis in Asian elephant was adapted and applied using RBPT, EDTA-TAT, and iELISA in the present study. For future evaluation, we recommended the use of a combination of serological tests with validation together with comparing by direct detection such as bacterial isolation to provide an appropriate brucellosis surveillance program in Asian elephants. In addition, the surveillance of stray dogs or multispecies habitation should be kept into considerations.

The Asiatic elephant (Elephas maximus indicus) is an endangered species that plays a critical role in wildlife conservation and landscape management across India. In many protected areas, captive elephants are indispensable for anti-poaching patrols, eco-tourism activities, human–wildlife conflict mitigation, and routine forest operations. Despite their importance, foot disorders remain among the most frequent health problems affecting their welfare, mobility, and overall working efficiency. Although captive elephants are widely used throughout the protected areas of Madhya Pradesh, no systematic, science-based assessment of foot affections had previously been undertaken. In this context, the present study evaluated 59 captive Asiatic elephants across the Kanha, Bandhavgarh, Panna, Pench, and Satpura Tiger Reserves to assess foot health and identify associated pathogens over a one-year period (April 2024–March 2025). Notably, Shigella flexneri was isolated from four foot-wound samples, with most cases detected in Bandhavgarh Tiger Reserve. The isolates were identified as gram-negative, non-motile rods exhibiting multidrug-resistant profiles, including resistance to Clindamycin, Penicillin-G, Gentamicin, Cefazolin and Cefoxitin, while retaining sensitivity to Ciprofloxacin, Ceftriaxone, Imipenem, Meropenem, and several other antibiotics. The calculated MAR index (0.129) suggests notable antimicrobial exposure within the system. To our knowledge, this is the first documented report of Shigella flexneri isolated from foot wounds in captive Asiatic elephants. This finding highlights the bacterium’s potential pathogenic role in promoting inflammation, tissue destruction, and delayed wound healing, underscoring the need for enhanced surveillance and improved foot-care management in captive elephant populations

Ethno-veterinary practices in Southern India for captive Asian elephant ailments

Parasites can influence the fitness of individuals particularly of small populations of endangered species. An island-wide, cross sectional, coprological survey was carried out from 03 January to 30 October 2015, to determine the gastrointestinal (GI) parasites of the Sri Lankan Elephant Elephas maximus maximus. Fresh fecal samples from wild, captive and semi-captive elephants were collected and analyzed using a modified salt floatation, Sheather’s sucrose floatation, direct iodine smears, and sedimentation methods. Species identification was done morphologically. Intensity of parasite infections was determined using McMaster technique. A total of 85 fecal samples (wild = 45, semi-captive = 20, captive = 20) were analysed; 58 (68.2%) samples were positive for GI parasites. Overall, helminth infections (60.0%) were more common than protozoan (37.6%) infections (Chi square test, χ2 = 8.499; p < 0.001). In the captive elephants, however, more protozoan infections were observed than helminthes, which could be due to anthelminthic treatment. A significantly higher prevalence of infection was observed in the wild elephants (93.3%) compared to semi-captive elephants (55.0%; χ2 = 13.516; p < 0.001) and captive elephants (25.0%; χ2 =32.289; p < 0.001) but there was no significant difference in the prevalence between captive and semi-captive elephants (χ2 =3.750; p = 0.053). Ten types of GI parasites were observed, nine of which were recorded in wild elephants. Among them the most common infection was strongyles (34.1%) with high intensity (440.1±295.2 EPG). Semi-captive elephants harbored five types of GI parasites, while captive elephants had only three types. One captive elephant at the Temple of the Tooth was infected with the tapeworm Anoplocephala sp. at low intensity of 50 EPG. Some of the GI parasites recorded are highly pathogenic while others are incidental.

We conducted a tuberculosis (TB) serosurveillance program of captive elephants in Nepaland compared hematology and biochemistry parameters between seropositive and seronegativeelephants. A total of 153 elephants (male=20, female=133) from four national parks weretested for TB using the ElephantTB STAT-PAK® Assay (ChemBio Diagnostic Systems,Inc., Medford, NY, USA). The mean reported age for 138 elephants was 38.5 years (range2–71 years). Seroprevalence for TB was 21.56% (33/153). The majority of seropositiveelephants were female (n=30) and from Chitwan National Park (n=29). The occurrence of TBseropositive cases in other more remote national parks suggests TB may be widespread amongthe captive elephant population of Nepal. Hematology and biochemistry analyses wereperformed on 13 and 22 seropositive elephants, respectively and, nine elephants from aseronegative TB herd for comparison. Hematology parameters (hemoglobin, packed cellvolume, platelet, white blood cells, and erythrocyte sedimentation rate) were comparablebetween the two groups. Total protein, globulin, and lactate dehydrogenase weresignificantly higher in seronegative elephants, and bilirubin was significantly higher inseropositive elephants whereas blood urea nitrogen, creatinine, glutamic oxaloacetictransaminase/aspartate aminotransferase (GOT/AST), glutamic pyruvic transaminase/alanineaminotransferase (GPT/ALT), gamma glutamyl transferase (GT), and albumin were notsignificantly different. The range of biochemical parameters that were significantlydifferent between seropositive and seronegative elephants had narrow ranges. Thus, thepotential of these parameters as a direct biomarker for TB diagnosis is limited based onthe findings in this study. We recommend including blood parameters in future TBsurveillance studies.

ABSTRACTElephant endotheliotropic herpesvirus (EEHV) can cause lethal hemorrhagic disease in juvenile Asian elephants, an endangered species. One hypothesis to explain this vulnerability of some juvenile elephants is that they fail to mount an effective T cell response to the virus. To our knowledge, there have been no studies of Asian elephant T cell responses to EEHV. To address this deficiency, we validated the gamma interferon (IFN-γ) enzyme-linked immunospot assay for tracking antigen-directed T cell activity by monitoring rabies-specific responses in vaccinated elephants. In addition, we generated monoclonal antibodies to Asian elephant CD4 and CD8 to facilitate phenotypic T cell profiling. Using these tools, we screened healthy elephants with a history of EEHV infection for reactivity against nine EEHV proteins whose counterparts in other herpesviruses are known to induce T cell responses in their natural hosts. We identified glycoprotein B (gB) and the putative regulatory protein E40 as the most immunogenic T cell targets (IFN-γ responses in five of seven elephants), followed by the major capsid protein (IFN-γ responses in three of seven elephants). We also observed that IFN-γ responses were largely from CD4+ T cells. We detected no activity against the predicted major immediate early (E44) and large tegument (E34) proteins, both immunodominant T cell targets in humans latently infected with cytomegalovirus. These studies identified EEHV-specific T cells in Asian elephants for the first time, lending insight into the T cell priming that might be required to protect against EEHV disease, and will guide the design of effective vaccine strategies.IMPORTANCE Endangered Asian elephants are facing many threats, including lethal hemorrhagic disease from elephant endotheliotropic herpesvirus (EEHV). EEHV usually establishes chronic, benign infections in mature Asian elephants but can be lethal to juvenile elephants in captivity and the wild. It is the leading cause of death in captive Asian elephants in North America and Europe. Despite the availability of sensitive tests and protocols for treating EEHV-associated illness, these measures are not always effective. The best line of defense would be a preventative vaccine. We interrogated normal healthy elephants previously infected with EEHV for T cell responses to nine EEHV proteins predicted to induce cellular immune responses. Three proteins elicited IFN-γ responses, suggesting their potential usefulness as vaccine candidates. Our work is the first to describe T cell responses to a member of the proposed fourth subfamily of mammalian herpesviruses, the Deltaherpesvirinae, within a host species in the clade Afrotheria. An EEHV vaccine would greatly contribute to the health care of Asian and African elephants that are also susceptible to this disease.

Elephant endotheliotropic herpesviruses (EEHVs) are important causes of death in both captive and wild Asian elephants (Elephas maximus). Nothing is known about the prevalence of EEHVs in wild or domestic elephants in China. To determine if EEHVs are present in elephants in China, 126 wild elephants from three populations and 202 captive individuals from zoos (n = 155) and the Wild Elephant Valley (n = 47) were screened using semi-nested polymerase chain reaction assays with EEHV-redundant and EEHV1/4/5-specific primers. EEHV1B and EEHV4 were detected in samples from both wild (EEHV1B:8/126; EEHV4:2/126) and captive (EEHV1B:5/155; EEHV4:9/155) elephants, while EEHV1A (six cases) and EEHV5 (one case) were only present in the captive elephants from the Wild Elephant Valley. EEHV1 was detected in blood and trunk and oral swabs; EEHV4 was detected in trunk and oral swabs as well as feces; EEHV5 was found in trunk and oral swabs. No significant age or sex association with EEHV1A, EEHV1B, or EEHV5 positivity was observed. An age association with EEHV4 positivity was found, with all unweaned elephants being EEHV4 positive, but an association with the sex of the elephant was not observed. These findings represent the first documentation of EEHV presence in captive and wild elephants in China. These findings also document EEHV1B and EEHV4 shedding in feces and demonstrate the utility of fecal screening as a tool for investigating EEHV4 infection in wild populations of elephants. It is recommended that EEHV testing be included in surveillance programs for captive and wild elephants in China.