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Background Approximately one-third of Asian elephants are managed under human care, participating in educational, cultural, religious, and tourist activities. Management conditions vary considerably among venues, raising questions about whether welfare needs are consistently being met, particularly for Asian tourist camp elephants. To evaluate the well-being of elephants engaged in tourist activities, an evidence-based tool is needed for routine assessments to identify potential welfare risks, aid in the development of better camp standards, and enable caretakers to address specific concerns. While many animal welfare tools exist, none have been designed to consider specific environments and management practices faced by elephants living and working in tourist camps. Methods Using direct observations and interviews, the Elephant Welfare Assessment Tool (EWAT) was developed for tourist camp elephants using the Five Domains Model as a framework. Measures were selected based on peer-reviewed literature, existing standards and guidelines, and opinions from animal welfare experts working with zoo and tourist camp elephants. The EWAT differs from existing tools by including criteria on work activities and restraint methods (e.g., chaining and ankus use), factors common in Asia but not often encountered by western zoo elephants. Measures were scored using a 0–2 Likert Scale. The tool was tested in Thailand and determined by calculating a content validity index (CVI) and conducting inter-rater and test-retest reliability tests. Results The initial tool included 18 animal-based and 21 resource-based measures across four domains: Nutrition (n = 5), Environment (n = 14), Health (n = 10), and Behavior and Mental State (n = 10). Index scores of content validity (CVI) (Item CVI (0.83), Scale CVI/Average (0.98), and Scale CVI/Universal (0.89)) were high. Measures scoring less than 0.83 were removed: the opportunity to mate, the mahout-elephant relationship, and mahout job satisfaction. The final tool consisted of 42 questions related to 36 measures, including 18 animal-based and 18 resource-based measures within the Nutrition (n = 5), Environment (n = 11), Health (n = 10), and Behavior and Mental State (n = 10) domains. Intraclass correlation coefficients (ICC) for inter-rater reliability (0.78–0.90, p < 0.05) and test-retest (0.77–0.91, p < 0.05) analyses conducted at two camps showed good agreement. Conclusions This new assessment tool (EWAT) is a context-specific, holistic method designed to offer a practical means of conducting individual and institutional-level assessments of elephant welfare in tourist camps. It is based on the Five Domains Model using reliable and validated animal- and resource-based measures, data collection through direct observation and interviews, and a numerical scoring system. The tool includes several criteria applicable to tourist rather than zoo venues to make it more relevant to the challenges faced by working elephants in Asia.

Abstract Gut microbiota play an important role in the health and disease of Asian elephants, however, its characteristics at each stage of life have not been thoroughly investigated in maintaining and regulating health of elephants. This study, therefore, aimed to characterize the profiles of the gut microbiota of captive Asian elephants from infants to the elderly. Gut microbiota were identified by 16S rRNA sequencing from the feces of captive Asian elephants with varying age groups, including infant calves, suckling calves, weaned calves, subadult and adult elephants, and geriatric elephants. The diversity of the gut microbiota was lowest in infants, stable during adulthood, and slightly decreased in the geriatric period. The gut microbiota of the infant elephants was dominated by milk-fermenting taxa including genus Bifidobacterium of family Bifidobacteriaceae together with genus Akkermansia. The fiber-fermenting taxa such as Lachnospiraceae_NK3A20_group were found to be increased in suckling elephants in differential abundance analysis by Analysis of Compositions of Microbiomes with Bias Correction (ANCOM-BC). The gut microbiota profiles after weaning until the adult period has been uniform as indicated by no significant differences in beta diversity between groups. However, the composition of the gut microbiota was found to change again in geriatric elephants. Understanding of the composition of the gut microbiota of captive Asian elephants at various life stages could be beneficial for promoting good health throughout their lifespan, as well as ensuring the welfare of captive elephants.

The abdominal testes of Asian elephants show normal spermatogenesis. Heat shock in cryptorchid testes elevates heat shock factor (HSF) expression, leading to germ cell apoptosis, while increased heat shock proteins (HSPs) levels provide protection. To investigate how heat shock affects elephant spermatogenic cells, focusing on heat shock-related molecules and the cell death mechanism, immunohistochemistry and TUNEL staining were employed to assess the immunoexpression of several heat shock-related molecules and the status of apoptosis in elephant fibroblasts (EF) induced by heat shock stimulus. Additionally, the immunoexpression of heat shock-related molecules and cell proliferation status in the elephant spermatogenic cells. Our finding indicated that heat shock-induced HSF1 immunoexpression in EF leads to apoptosis mediated by T-cell death-associated gene 51 (TDAG51) while also upregulating HSP70 to protect damaged cells. In elephant spermatogenic cells, immunostaining revealed a predominance of proliferating cell nuclear antigen (PCNA)-positive cells with minimal TDAG51- and TUNEL-positive cells, suggesting active proliferation and apoptosis suppression during normal spermatogenesis in the abdominal testis. Interestingly, spermatogonia co-immunoexpressed HSF1 and HSP90, potentially reducing apoptosis through protective mechanisms different from those observed in other mammals. Spermatogenic cells did not show immunolocalisation of HSP70, and hence, it may not contribute to protecting the spermatogonia from heat shock because the transcriptional activity of HSF1 is suppressed by HSP90A binding. This study provides insight into the specific heat shock response and defence mechanisms in elephant spermatogenic cells and may contribute to our understanding of species-specific adaptation to environmental stresses of the testis.

Welfare is a fundamental aspect of animal management and conservation. In light of growing public awareness and welfare concerns about captive elephants, there is an urgent need for comprehensive, globally coordinated efforts for Asian elephants (Elephas maximus) that participate in religious, logging, or tourist activities in range countries where the majority reside, and where welfare issues have been identified but not addressed. This review provides a comparative analysis of available animal assessment tools. Each offers distinct features for assessment that allow institutions to select criteria for specific needs and available resources. Most are applied to general animal welfare assessments, although some are tailored to particular species, including elephants. The tools span diverse formats, from digital to primarily paper-based assessments. Assessments operate at individual and institutional levels and across multiple welfare domains. Methodologies rely on keeper ratings or expert evaluations, incorporate numerical scoring and Likert scales for welfare grading, and encompass inputs including behaviors, health, and physiological indicators. For tourist camp elephants, one challenge is that the tools were developed in zoos, which may or may not have application to non-zoological settings. Digital tools and assessment methodologies such as keeper ratings face logistical challenges when applied across tourist venues. As with any tool, reliability, validity, and repeatability are essential and must address the unique welfare challenges of diverse captive settings. We propose that a holistic, context-specific, evidence-based, and practical tool be developed to ensure high elephant welfare standards in non-zoological facilities throughout Asia.

The international travel ban instituted by the Thai government in March 2020 in response to the COVID-19 pandemic greatly affected how tourist camp elephants were managed, with reductions in exercise opportunities, longer chaining hours, and diminished food provisioning. This study was conducted to determine how those changes affected health and welfare biomarkers in individual elephants over the 2 years of the countrywide lockdown (April 2020–April 2022). Blood and fecal samples were collected from 58 elephants at six camps (monthly in Year 1, quarterly in Year 2) and analyzed for stress biomarkers — fecal glucocorticoid metabolites (fGCM), serum oxidative stress [malondialdehyde (MDA) and 8-hydroxy-2′-deoxyguanosine (8-OHdG)], and stress leukograms. Overall, fGCM concentrations increased within the first few months and remained higher than pre-COVID levels, as did the H/L ratio, a measure affected by cortisol. Serum 8-OHdG, an indicator of DNA oxidative damage, also increased over time, while monocytosis and lymphopenia further suggested alterations in immune function as a result of stress. By contrast, another marker of oxidative stress, serum MDA, declined, possibly in response to reduced roughage and supplement intake. A notable finding was a seasonal pattern of fGCM that was significantly different from previous studies. Whereas higher fGCM during the rainy season were observed in this study, previously, concentrations were highest during the winter, high tourist season. Thus, ironically, both the presence and absence of tourists have been associated with increased fGCM concentrations, albeit for different reasons. Camp management factors negatively affecting stress outcomes included shorter chain lengths, longer chain hours, lack of exercise, and reduced roughage and supplements. Overall, it was clear that camps struggled to maintain adequate care for elephants during the COVID-19 pandemic, highlighting the importance of tourist income and need for contingency plans to cope with potential future disruptions to tourism.

The international travel ban initiated in March 2020 due to the COVID-19 pandemic greatly affected how captive elephants were managed in Thailand. A lack of tourists and associated income meant elephants were chained longer with reduced food provisions, had fewer mahouts, and limited exercise like riding, which likely affected health and welfare. Fifty-eight elephants from six tourist camps were assigned a body condition score (BCS) and blood samples were collected monthly for 2 years during the travel ban to measure: (1) muscle enzymes [creatine kinase (CK), aspartate aminotransferase (AST)]; (2) liver enzymes [aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT)]; (3) lipids [total cholesterol (TC), triglycerides (TG), low (LDL) and high (HDL) density lipoproteins]; and metabolic function [glucose, insulin, fructosamine]. Serum CK concentrations were lower at the end of the study, possibly due to no tourist activities like riding. Changes in liver function included increased AST and ALP, also possibly due to physical inactivity. Feeding less bananas and sugar cane was associated with fewer elephants in the obese category and lower TG concentrations. However, increases in glucose, insulin and fructosamine were observed as local people returned to feed elephants after lifting travel restrictions. In sum, changes in several health biomarkers were observed in association with restricted activities and food resources. Camps need better plans to meet the health and welfare needs of elephants during any future disruptions to the tourism industry, although reduced feeding of sweet treats appeared to have positive effects on body condition and metabolic function.

IntroductionOxidative stress refers to an imbalance between oxidant and antioxidant activity and accumulation of reactive oxygen species, which can have detrimental effects on animal health. Annual fluctuations in oxidative stress status can occur, increasing disease susceptibility during certain time periods. However, a full understanding of factors related to oxidative stress in Asian elephants and how to mitigate the negative consequences is lacking.MethodsThis study measured six serum oxidative stress markers [reactive oxygen species (ROS), malondialdehyde (MDA), 8-hydroxydeoxyguanosine (8-OHdG), albumin, glutathione peroxidase (GPx), and catalase] and two stress markers [serum cortisol and fecal glucocorticoid metabolites (fGCM)] in 23 captive Asian elephants in Thailand over a 12 months period to examine relationships with age and season.ResultsSeasonal variations were observed, with several markers exhibiting significantly higher concentrations in the summer (ROS, MDA, 8-OHdG, albumin) and lower values during the rainy/winter seasons (MDA, 8-OHdG, albumin, catalase). By contrast, GPx was the only marker to be highest during the rainy season. For the stress markers, higher fGCM concentrations were noted during the rainy season, which contrasts with earlier studies showing more activity in the winter (tourist season). Positive correlations were found between the temperature-humidity index and ROS, GPx, and fGCM, while a negative correlation was observed with serum albumin. Elephant endotheliotropic herpesvirus (EEHV) shedding events were associated with higher concentrations of ROS and MDA. A moderate negative correlation was observed between 8-OHdG and the PCR threshold cycle of EEHV shedding (Ct), indicating DNA damage may be involved in EEHV shedding in elephants.DiscussionResults revealed significant age and seasonal effects on several oxidative stress markers, indicating those factors should be considered in study design and data interpretation. There also may be physiological adaptations in oxidative stress conditions in relation to environmental changes that could impact health outcomes.

Three cases of double infection with elephant endotheliotropic herpesvirus (EEHV) types 1A and 4 in captive Asian elephants are presented. The first calf was a 4-year-old female that showed initial signs of lethargy and depression. The second calf was a 6-year-old female that displayed signs of depression and diarrhea and died within 48 h of the start of supportive treatment. The third was a 2-year-old male that died suddenly while living with the herd. Necropsies were performed in the first and second elephants, while only a tongue sample was collected from the third calf. EEHV infection was confirmed via quantitative PCR (qPCR) and gene sequencing, revealing double subtypes of EEHV1A and -4 infections. This study describes the hematological and pathological characteristics within the host following double EEHV infection.

Background Elephant endotheliotropic herpesvirus causes a hemorrhagic disease (EEHV-HD) that is a major cause of death in juvenile Asian elephants with EEHV1 and EEHV4 being the most prevalent. Aim To perform a retrospective clinical data analysis. Methods Records of a total of 103 cases in Thailand confirmed by polymerase chain reaction (PCR) on blood and/or tissue samples. Results The severity of clinical signs varied among EEHV subtypes. EEHV1A was the most prevalent with 58%, followed by EEHV4 with 34%, EEHV1B with 5.8% and EEHV1&4 co-infection with 1.9%. Overall case fatality rate was 66%. When compared among subtypes, 100% case fatality rate was associated with EEHV1&4 co-infection, 83% with EEHV1B, 75% with EEHV1A, and the lowest at 40% for EEHV4. Calves 2- to 4-year old were in the highest age risk group and exhibited more severe clinical signs with the highest mortality. Majority of cases were found in weaned or trained claves and higher number of cases were observed in rainy season. A gender predilection could not be demonstrated. Severely affected elephants presented with thrombocytopenia, depletion of monocytes, lymphocytes and heterophils, a monocyte:heterophil (M:H) ratio lower than 2.37, hypoproteinemia (both albumin and globulin), severe grade of heterophil toxicity, and low red blood cell counts and pack cell volumes. Survival was not affected by antiviral drug treatment in the severely compromised animals. Conclusion Early detection by laboratory testing and aggressive application of therapies comprising of supportive and anti-viral treatment can improve survival outcomes of this disease.

The COVID-19 pandemic has had a significant impact on the tourism industry, especially in Thailand. Starting in April 2020, the Thai government banned international travel and all elephant tourist camps closed. A wide variety of management changes were implemented because of the lack of income from tourists. This study surveyed 30 camps that cared for >400 elephants in northern Thailand to obtain information on camp, elephant, and mahout management during the COVID-19 pandemic from April 2020 to 2022 compared to the year before. The survey consisted of questionnaires that interviewed elephant camp owners, managers, veterinarians, and mahouts, and captured information on changes in camp operations, including numbers of tourists, elephants and mahouts, elephant and mahout activities, and veterinary care. Results revealed significant changes in camp structure, elephant work activities and general care. Staff layoffs led to a decrease in the ratio of mahouts to elephants from 1:1 to 1:2. Elephant activities, distance walked, and amounts of food were reduced when compared to pre-COVID-19, while chain hours were increased due to reduced activity. Overall, the COVID-19 crisis altered elephant management significantly, potentially affecting animal welfare resulting from changes in nutrition, health, exercise, and numbers of mahouts. We hope to use these data to develop better management plans and guidelines for elephant camps in Thailand so they can cope with the current and potential imminent pandemics that result in decreased tourism income. A follow-up study will measure health and welfare markers in relation to COVID-19 induced changes to determine if any camps adapted management to still meet elephant health and welfare needs, and could serve as models for responding to future pandemics.

Abstract Elephant endotheliotropic herpesvirus-hemorrhagic disease (EEHV-HD) is an acute fatal disease in elephants. Despite the fact that the underlying pathogenesis of EEHV-HD has been proposed, it remains undetermined as to what mechanisms drive these hemorrhagic and edematous lesions. In the present study, we have investigated and explained the pathogenesis of acute EEHV-HD using blood profiles of EEHV-HD and EEHV-infected cases, hematoxylin and eosin (H&E) stain, special stains, immunohistochemistry, quantitative polymerase chain reaction (PCR) and reverse transcriptase polymerase chain reaction (RT-PCR). It was found that EEHV genomes were predominantly detected in various internal organs of EEHV-HD cases. Damage to endothelial cells, vasculitis and vascular thrombosis of the small blood vessels were also predominantly observed. Increases in platelet endothelial cell adhesion molecules-1 (PECAM-1)- and von Willebrand factor (vWF)-immunolabeling positive cells were significantly noticed in injured blood vessels. The expression of pro-inflammatory cytokine mRNA was significantly up-regulated in EEHV-HD cases when compared to EEHV-negative controls. We have hypothesized that this could be attributed to the systemic inflammation and disruption of small blood vessels, followed by the disseminated intravascular coagulopathy that enhanced hemorrhagic and edematous lesions in EEHV-HD cases. Our findings have brought attention to the potential application of effective preventive and therapeutic protocols to treat EEHV infection in Asian elephants.

Oxidative stress is a pathological condition that can have adverse effects on animal health, although little research has been conducted on wildlife species. In this study, blood was collected from captive Asian elephants for the assessment of five serum oxidative status markers (reactive oxygen species (ROS) concentrations; malondialdehyde, MDA; albumin; glutathione peroxidase, GPx; and catalase) in healthy (n = 137) and sick (n = 20) animals. Health problems consisted of weakness, puncture wounds, gastrointestinal distress, eye and musculoskeletal problems, and elephant endotheliotropic herpesvirus hemorrhagic disease (EEHV-HD). Fecal samples were also collected to assess glucocorticoid metabolites (fGCMs) as a measure of stress. All data were analyzed in relation to age, sex, sampling season, and their interactions using generalized linear models, and a correlation matrix was constructed. ROS and serum albumin concentrations exhibited the highest concentrations in aged elephants (>45 years). No sex differences were found for any biomarker. Interactions were observed for age groups and seasons for ROS and catalase, while GPx displayed a significant interaction between sex and season. In pairwise comparisons, significant increases in ROS and catalase were observed in summer, with higher ROS concentrations observed only in the adult female group. Lower catalase activity was exhibited in juvenile males, subadult males, adult females, and aged females compared to subadult and adult elephants (males and females) in winter and the rainy season. There was a positive association between catalase activity and fGCMs (r = 0.23, p < 0.05), and a number of red blood cell parameters were positively associated with several of these biomarkers, suggesting high oxidative and antioxidative activity covary in red cells (p < 0.05). According to health status, elephants with EEHV-HD showed the most significant changes in oxidative stress markers, with MDA, GPx, and catalase being higher and albumin being lower than in healthy elephants. This study provides an analysis of understudied health biomarkers in Asian elephants, which can be used as additional tools for assessing the health condition of this species and suggests age and season may be important factors in data interpretation.

A therapeutic regimen that includes antiviral drugs is critical for the survival of Asian elephant (Elephas maximus) calves infected with elephant endotheliotropic herpesvirus hemorrhagic disease (EEHV-HD), with acyclovir showing considerable promise. The purpose of this study was to determine the pharmacokinetics and bioavailability of acyclovir following intravenous (IV) and oral (PO) administration in Asian elephants. A single dose of acyclovir (15 mg/kg, IV or 45 mg/kg, PO) was administered to four healthy elephant calves, with a minimum 2-week washout period between treatments. Serial plasma samples were collected after each injection for acyclovir analysis using a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) technique. Maximum plasma acyclovir concentrations were 27.02 ± 6.79 µg/mL at 0.94 ± 0.31 h after IV administration, and 1.45 ± 0.20 µg/mL at 3.00 ± 0.70 h after PO administration. The half-life of the elimination phase (T1/2) was 5.84 ± 0.74 and 8.74 ± 2.47 h after IV and PO administration, respectively. After IV administration, acyclovir concentrations were higher than the half-maximal inhibitory concentration (IC50) of those found for herpes simplex virus (HSV) 1 and 2 in humans, and equid alpha herpesvirus-1 (EHV-1) for at least 12 h. By contrast, the bioavailability of oral administration was low, only 6.03 ± 0.87%, so higher doses by that route likely are needed to be effective. Due to the high concentration of plasma acyclovir after IV administration, the dose may need to be adjusted to prevent any negative side effects.

This study investigated the sedative effects of dexmedetomidine in Asian elephants. We hypothesized that 2 µg/kg dexmedetomidine would provide sufficient standing sedation. A crossover design study was performed in three Asian elephants. Each elephant was assigned to 1 of 3 treatment groups—1 (D1), 1.5 (D1.5) or 2 (D2) µg/kg dexmedetomidine (intramuscular injection, IM) with a two-week ‘washout period’ between doses. Elephants were monitored for 120 min. At 120 min (Ta), atipamezole was administered IM. Sedation and responsiveness scores were evaluated. Physiological parameters (pulse rate, respiratory rate, and %SpO2) and clinical observations were monitored during the study and for 3 days post drug administration. D2 provided the longest sedation (approximately 70 min), compared to D1 and D1.5. After Ta, each elephant’s sedative stage lessened within 10–15 min without complications. No significant abnormal clinical observations were noted throughout and during the 3-days post study period. These data suggest that a single 2 µg/kg IM dexmedetomidine injection provides sufficient standing sedation for approximately 70 min in Asian elephants.

Abstract Background Osteoarthritis (OA), the most common form of arthritic disease, results from destruction of joint cartilage and underlying bone. It affects animals, including Asian elephants (Elephas maximus) in captivity, leading to joint pain and lameness. However, publications regarding OA pathogenesis in this animal are still limited. Therefore, this study aimed to investigate the effect of proinflammatory cytokines, including interleukin-1 beta (IL-1β), IL-17A, tumor necrosis factor-alpha (TNF-α), and oncostatin M (OSM), known mediators of OA pathogenesis, and lipopolysaccharides on the expression of cartilaginous degrading enzymes, matrix metalloproteinase (MMP)-3 and MMP-13, in elephant articular chondrocytes (ELACs) cultures. Anti-arthritic drugs and the active compounds of herbal plants were tested for their potential attenuation against overproduction of these enzymes. Results Among the used cytokines, OSM showed the highest activation of MMP3 and MMP13 expression, especially when combined with IL-1β. The combination of IL-1β and OSM was found to activate phosphorylation of the mitogen-activated protein kinase (MAPK) pathway in ELACs. Lipopolysaccharides or cytokine-induced expressions were suppressed by pharmacologic agents used to treat OA, including dexamethasone, indomethacin, etoricoxib, and diacerein, and by three natural compounds, sesamin, andrographolide, and vanillylacetone. Conclusions Our results revealed the cellular mechanisms underlying OA in elephant chondrocytes, which is triggered by proinflammatory cytokines or lipopolysaccharides and suppressed by common pharmacological or natural medications used to treat human OA. These results provide a more basic understanding of the pathogenesis of elephant OA, which could be useful for adequate medical treatment of OA in this animal.

Non-thermal plasma (NTP) is being developed for a wide-range of medical applications such as improvement of wound healing, elimination of infective microorganisms, and treatment of cancer. This study investigated the effect of culture medium exposed to NTP on the proliferation in-vitro of skin fibroblasts from Asian elephants. Dulbecco"s Modified Eagle"s Medium (DMEM) was used as culture medium and was exposed to NTP with three different intensities. The NTP reactive species Nitrite (NO2-) was measured in the treated medium before addition to cells. Fibroblasts were incubated for 24 h with NTP-treated complete medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% antibiotic/antimycotic. Cell proliferation, the number of cells and viability rate were analysed using flow cytometry 24, 48 and 72 h after the start of the incubation. The proliferation rate of fibroblasts incubated with NTP treated medium was significantly higher (P

Gait analysis is a method of gathering quantitative information to assist in determining the cause of abnormal gait for the purpose of making treatment decisions in veterinary medicine. Recent technology has offered the wearable wireless sensor of an inertial measurement unit (IMU) for determining gait parameters. This study proposed the use of a triaxial IMU, comprising an accelerometer, a gyroscope, and a magnetometer, for detecting three-dimensional limb segment motion (XYZ axis) during the gait cycle in Asian elephants. A new algorithm was developed to estimate the kinematic parameter that represents each limb segment of the forelimbs and hindlimbs while walking at a comfortable speed. For future use, this study aimed to create a new prototype of the IMU with a configuration that is tailored to the elephant and apply machine learning in an effort to achieve greater precision.

Disease caused by elephant endotheliotropic herpesvirus (EEHV) infection is the most highly fatal hemorrhagic disease in Asian elephant calves worldwide. To date, adult elephants that have been infected with EEHV have predominantly displayed mild clinical signs, while they are believed to serve as EEHV shedders to other elephants. Hence, the diagnostic tools employed for monitoring EEHV-active infection are considered vitally important. In this study, partial EEHV-DNA polymerase (DNApol) nonstructural proteins (NSPs) were used to detect anti-EEHV antibodies through the use of an in-house indirect enzyme-linked immunosorbent assay (ELISA). The results were then compared to those obtained from a PCR test. In this study, a total of 175 serum samples were collected from Asian elephants living in elephant camps located in Chiang Mai and Lampang Provinces, Thailand. The elephants were aged between 2 and 80 years old. The overall percentages of positive samples by the PCR and EEHV-DNApol ELISA tests were 4% (21/175) and 12% (21/175), respectively. The ELISAs demonstrated values of 77.9% (95% posterior probability interval (PPI) = 52.5–95%) sensitivity and 87.7% (PPI = 82.5–91.9%) specificity, respectively. Accordingly, the sera obtained from the elephants exhibiting no clinical signs of EEHV infection, and those who were negative according to PCR tests, revealed a value of 14% seropositivity for EEHV-DNApol. Our results indicate that these asymptomatic, active EEHV-infected elephants could likely serve as a source of EEHV shedding within elephant herds. Consequently, the developed EEHV-DNApol NSPs-based ELISA test employed in the present study may be of use for routine monitoring and identification of EEHV shedders in elephant herds, and could be an alternative diagnostic tool for EEHV detection in Asian elephants.

Currently, species identification of stranded marine mammals mostly relies on morphological features, which has inherent challenges. The use of genetic information for marine mammal species identification remains limited, therefore, new approaches that can contribute to a better monitoring of stranded species are needed. In that context, the ISSR-HRM method we have proposed offers a new approach for marine mammal species identification. Consequently, new approaches need to be developed to identify individuals at the species level. Eight primers of the ISSR markers were chosen for HRM analysis resulting in ranges of accuracy of 56.78–75.50% and 52.14–75.93% in terms of precision, while a degree of sensitivity of more than 80% was recorded when each single primer was used. The ISSR-HRM primer combinations revealed a success rate of 100% in terms of discrimination for all marine mammals included in this study. Furthermore, ISSR-HRM analysis was successfully employed in determining marine mammal discrimination among varying marine mammal species. Thus, ISSR-HRM analysis could serve as an effective alternative tool in the species identification process. This option would offer researchers a heightened level of convenience in terms of its performance and success rate. It would also offer field practice to veterinarians, biologists and other field-related people a greater degree of ease with which they could interpret results when effectively classifying stranded marine mammals. However, further studies with more samples and with a broader geographical scope will be required involving distinct populations to account for the high degree of intraspecific variability in cetaceans and to demonstrate the range of applications of this approach.

The aim of this study was to examine relationships among serum progestagens, cortisol, and prolactin in pregnant and normal cycling Asian elephants living in tourist camps in northern Thailand. Samples were collected twice a month for 22 months from nine elephants. Of those, four were pregnant (24.3 ± 2.9 years of age; range 21–28 years) and five (20.2 ± 9.6 years; range 8–34 years) exhibited normal ovarian cycles based on serum progestagen analyses. Gestation was divided into three periods: 1st (week 1–31), 2nd (week 32–62), and 3rd (week 63 to parturition), while the estrous cycle was divided into the follicular and luteal phases. Serum progestagens were higher during the luteal phase of the cycle (p < 0.003), whereas cortisol and prolactin were similar. In pregnant elephants, there were no differences in serum progestagens or cortisol concentrations across the three gestational periods, whereas prolactin concentrations increased significantly during the 2nd and 3rd periods (p < 0.0001). By contrast, prolactin concentrations in nonpregnant elephants were consistently low throughout the ovarian cycle. In one cycling female, prolactin concentrations were similar to pregnant elephants, perhaps because she was an allomother to two calves. Another cycling female exhibited consistently elevated cortisol concentrations, 5 to 10 times higher than the other elephants. There were no correlations between serum progestagens, cortisol, and prolactin throughout gestation; however, serum progestagens and cortisol were positively related in cycling elephants (r = 0.386, p < 0.001). From our results, there were a number of individual differences in reproductive hormonal patterns, so it is important to develop personalized monitoring programs for each elephant to enhance breeding success and create sustaining captive populations of elephants in Asia.

The identification of differing physical characteristics of dogs is an uncomplicated and straightforward way to categorize dog breeds. However, many dog owners and veterinarians still struggle to distinguish between pure breed and mixed variations in certain breeds of dogs. Presently, the absence of the tools and methods needed to confirm a pure breed dog is a significant problem since the only method available to validate pure or mongrel breeds is the official pedigree system. Inter-simple sequence repeat markers have been successfully used to assess genetic variations and differentiations. Notably, inter-simple sequence repeat markers coupled with high resolution melting analysis were effectively used for the breed identification of 43 breeds of dogs (total 463 dogs). The 10 primers chosen for analysis resulted in a range of 31–78.6% of breed discrimination when using one primer, while a combination of two primers was able to successfully discriminate between all of the 43 dog breeds (100%). Shannon’s index information (I = 2.586 ± 0.034) and expected heterozygosity (He = 0.908 ± 0.003) indicated a high level of genetic diversity among breeds. The fixation index (Fst) revealed a value of 10.4%, demonstrating that there was a high level of genetic subdivision between populations. This study showed that inter-simple sequence repeat marker analysis was effective in demonstrating high genetic diversity among varying breeds of dogs, while a combination of Inter-simple sequence repeat marker analysis and high resolution melting analysis could provide an optional technique for researchers to effectively identify breeds through genetic variations.

The Asian elephant population is continuously declining due to several extrinsic reasons in their range countries, but also due to diseases in captive populations worldwide. One of these diseases, the elephant endotheliotropic herpesvirus (EEHV) hemorrhagic disease, is very impactful because it particularly affects Asian elephant calves. It is commonly fatal and presents as an acute and generalized hemorrhagic syndrome. Therefore, having reference values of coagulation parameters, and obtaining such values for diseased animals in a very short time, is of great importance. We analyzed prothrombin time (PT), activated partial thromboplastin time (aPTT), and fibrinogen concentrations using a portable and fast point-of-care analyzer (VetScan Pro) in 127 Asian elephants from Thai camps and European captive herds. We found significantly different PT and aPTT coagulation times between elephants from the two regions, as well as clear differences in fibrinogen concentration. Nevertheless, these alterations were not expected to have biological or clinical implications. We have also sequenced the coagulation factor VII gene of 141 animals to assess the presence of a previously reported hereditary coagulation disorder in Asian elephants and to investigate the presence of other mutations. We did not find the previously reported mutation in our study population. Instead, we discovered the presence of several new single nucleotide polymorphisms, two of them being considered as deleterious by effect prediction software.

Background: Elephants in Thailand have changed their roles from working in the logging industry to tourism over the past two decades. In 2020, there were approximately 2700 captive elephants participating in activities such as riding and trekking. During work hours, riding elephants carry one or two people in a saddle on the back with a mahout on the neck several hours a day and over varying terrain. A concern is that this form of riding can cause serious injuries to the musculoskeletal system, although to date there have been no empirical studies to determine the influence of weight carriage on kinematics in elephants. Methods: Eight Asian elephants from a camp in Chiang Mai Province, Thailand, aged between 21 and 41 years with a mean body mass of 3265 ± 140.2 kg, were evaluated under two conditions: walking at a normal speed without a saddle and with a 15% body mass load (saddle and two persons plus additional weights). Gait kinematics, including the maximal angles of fore- and hindlimb joints, were determined using a novel three-dimensional inertial measurement system with wireless sensors. Results: There were no statistical differences between movement angles and a range of motion of the fore- and hindlimbs, when an additional 15% of body mass was added. Conclusion: There is no evidence that carrying a 15% body mass load causes significant changes in elephant gait patterns. Thus, carrying two people in a saddle may have minimal effects on musculoskeletal function. More studies are needed to further test longer durations of riding on different types of terrain to develop appropriate working guidelines for captive elephants. Nevertheless, elephants appear capable of carrying significant amounts of weight on the back without showing signs of physical distress.

Elephant endotheliotropic herpesvirus hemorrhagic disease (EEHV-HD) is a virulent disease that causes severe hemorrhage and sudden death in Asian elephant calves. A change in hematology profiles is one indicator of infection before clinical signs appear; however, to be effective, individual baselines and age-matched reference values are needed. Stress has been speculated to be a factor in clinical EEHV cases, but relationships have not been demonstrated empirically. This study evaluated blood hematology and several stress response markers—salivary cortisol, fecal glucocorticoid metabolites (FGM), salivary Immunoglobulin A (SIgA), and fecal IgA (FIgA) in samples collected for 1 year from three healthy calves with no EEHV history (non-EEHV), and six that had previously been infected, developed clinical signs and survived (prior-EEHV). Hematology values between non-EEHV and prior-EEHV elephants were not different and within published reference ranges. Concentrations of salivary cortisol, FGM, SIgA, and FIgA also were variable and showed seasonal differences, but no relationships to prior EEHV status. One of the prior EEHV calves became re-infected, developed hemorrhagic disease (HD), and died during the study period. That calf exhibited lymphocytopenia, monocytopenia, and thrombocytopenia. Additionally, all stress biomarker concentrations were lower in the 12 days before viremia was observed. Thus, as in other studies, changes in hematology occur with EEHV infection, while preliminary data in one calf suggests that stress-response measures might also be informative and should be studied further.

The aims of this study were to investigate the coding sequence and the deduced amino acid sequence of Asian elephant"s biglycan gene as well as its expression in different tissues and conditions using wound healing as a model. The results showed that Asian elephant biglycan coding sequence was 1,110 base pair (bp) long (accession number: JQ753329), encoding 369 amino acids. The coding and amino acid sequences between Asian and African elephants revealed 99% and 98% similarity, respectively. The conserved domains of biglycan protein were also observed. In addition, its expression was found in 15 tissues with a predominant expression in cartilage and spleen. For expression analysis in the wound healing process, it was found that the level of biglycan mRNA was influenced by many factors, including age, type of wound and stage of wound healing.

Background Variation in management across elephant camps likely has differential effects on the well-being of elephants. Methods This study calculated body condition, foot health and skin wound scores (WSs) for 122 elephants from 15 elephant camps in Chiang Mai province, and examined relationships to management factors using a multi-variable modeling approach. Results The majority of elephants had high body condition scores (BCS) indicative of being overweight or obese, mild foot problems, but few visible wounds. Females had higher BCSs than males, as did elephants provided a water source at night. Increasing age was associated with higher foot and WSs. Higher WSs were observed in about a quarter of the cases where mahouts carried a hook. Wounds related to saddle riding were rare. Elephants that rested on sand floors at night had a decreased risk of high WSs compared to elephants that rested on compact dirt floors. Discussion Findings emphasize the need for elephant camps to adjust management activities that negatively affect body condition (e.g., feeding too many sweet treats), foot health (e.g., hard substrates) and wounding (e.g., misuse of equipment) to improve health and welfare of this population.

This study investigated how camp management and tourist activities affect body condition, adrenocortical function, lipid profiles and metabolic status in female tourist elephants. We compared twice monthly serum insulin, glucose, fructosamine, total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL), high density lipoprotein (HDL), and fecal glucocorticoid metabolite (FGM) concentrations to body condition scores (BCS) at five camps with different management styles (e.g., tourist activities, work type, diet) between the High (November-February) and Low (March-October) tourist seasons. There were significant camp effects on health parameters, with BCS, TC, HDL, insulin and glucose being among the highest, and G:I being the lowest (less heathy) in elephants at an observation camp compared to those at camps where elephants received exercise by providing rides to tourists. Differences between High and Low tourist season months also were found for all measures, except TG and FGM concentrations. Both work time and walking distance were negatively correlated to glucose, fructosamine and insulin, while walking distance was negatively related to FGM concentrations. By contrast, positive associations were found between tourist number and BCS, TG, and insulin, perhaps related to tourists feeding elephants. Quantity of supplementary diet items (e.g., bananas, sugar cane, pumpkin) were positively correlated with FGM concentrations, glucose, fructosamine, and insulin. This study provides evidence that body condition, adrenal activity, metabolic markers, and lipid profiles in captive elephants may be affected by visitor numbers, work activities, and the amount of supplementary foods offered by tourists. Some activities appear to have negative (e.g., feeding), while others (e.g., exercise) may have positive effects on health and welfare. We conclude that camps adopting a more hands-off approach to tourism need to ensure elephants remain healthy by providing environments that encourage activity and rely on more natural diets or foraging.

We previously found relationships between body condition and physiological function affecting health and welfare of female tourist camp elephants in Thailand, and used that approach to conduct a similar study of bull elephants in the same camps (n = 13). A body condition score (BCS) was done every other month, and fecal glucocorticoid metabolite (FGM) concentrations were measured twice monthly for 1 year. Effects of season, camp management and tourist activity on lipid profiles [total cholesterol (TC), low density lipoproteins (LDL), high density lipoproteins (HDL), triglycerides (TG)] and metabolic factors [insulin, glucose, fructosamine, glucose to insulin ratio (G:I)] were determined and correlated to measures of body condition, testosterone and FGM. Positive correlations were found between BCS and TG, between FGM and TG, HDL and glucose, and between testosterone and HDL, whereas BCS and testosterone were negatively associated with the G:I. There was a significant positive relationship between FGM and testosterone. Elevated FGM concentrations were associated with altered lipid and metabolic profiles and were higher in winter compared to summer and rainy seasons. Insulin and glucose levels were higher, while the G:I was lowest in the winter season. Strong positive associations were found between TC and HDL, LDL and HDL and glucose, and glucose and insulin. By contrast, negative relationships were found between the G:I and HDL and glucose, and between insulin and G:I. Differences also were found between High and Low tourist season months for FGM, insulin, and G:I. Last, there was notable variation among the camps in measured parameters, which together with tourist season effects suggests camp management may affect physiological function and welfare; some negatively like feeding high calorie treats, others positively, like exercise. Last, compared to females, bull elephants appear to be in better physical health based on normal BCSs, lower insulin levels and higher G:I ratios.

Elephant camps are among the most popular destinations in Thailand for tourists from many countries. A wide range of management strategies are used by these camps, which can have varied impacts on health and welfare of elephants. The objectives of this study were to examine relationships between FGM (fecal glucocorticoid metabolite) concentrations and camp management factors (work routine, walking, restraint, rest area, foraging), and to other welfare indicators (stereotypic behaviors, body condition, foot health, and skin wounds). Data were obtained on 84 elephants (18 males and 66 females) from 15 elephant camps over a 1-year period. Elephants were examined every 3 months and assigned a body condition score, foot score, and wound score. Fecal samples were collected twice monthly for FGM analysis. Contrary to some beliefs, elephants in the observation only program where mahouts did not carry an ankus for protection had higher FGM concentrations compared to those at camps that offered riding with a saddle and shows. Elephants that were tethered in the forest at night had lower FGM concentrations compared to elephants that were kept in open areas inside the camps. There was an inverse relationship between FGM concentrations and occurrence of stereotypy, which was not anticipated. Thus, assessing adrenal activity via monitoring of FGM concentrations can provide important information on factors affecting the well-being of elephants. Results suggest that more naturalistic housing conditions and providing opportunities to exercise may be good for elephants under human care in Thailand, and that a no riding, no hook policy does not necessarily guarantee good welfare.

Elephant endotheliotropic herpesvirus-hemorrhagic disease (EEHV-HD) is the primary cause of acute, highly fatal, hemorrhagic diseases in young Asian elephants. Although monocytopenia is frequently observed in EEHV-HD cases, the role monocytes play in EEHV-disease pathogenesis is unknown. This study seeks to explain the responses of monocytes/macrophages in the pathogenesis of EEHV-HD. Samples of blood, frozen tissues, and formalin-fixed, paraffin-embedded (FFPE) tissues from EEHV1A-HD, EEHV4-HD, co-infected EEHV1A and 4-HD, and EEHV-negative calves were analyzed. Peripheral blood mononuclear cells (PBMCs) from the persistent EEHV4-infected and EEHV-negative calves were also studied. The results showed increased infiltration of Iba-1-positive macrophages in the inflamed tissues of the internal organs of elephant calves with EEHV-HD. In addition, cellular apoptosis also increased in the tissues of elephants with EEHV-HD, especially in the PBMCs, compared to the EEHV-negative control. In the PBMCs of persistent EEHV4-infected elephants, cytokine mRNA expression was high, particularly up-regulation of TNF-α and IFN-γ. Moreover, viral particles were observed in the cytoplasm of the persistent EEHV4-infected elephant monocytes. Our study demonstrated for the first time that apoptosis of the PBMCs increased in cases of EEHV-HD. Furthermore, this study showed that monocytes may serve as a vehicle for viral dissemination during EEHV infection in Asian elephants.

Elephant endotheliotropic herpesvirus (EEHV) infection is known to cause acute fatal hemorrhagic disease, which has killed many young Asian elephants (Elephas maximus). Until recently, in vitro isolation and propagation of the virus have not been successful. This study aimed to isolate and propagate EEHV using continuous cell lines derived from human and/or animal origins. Human cell lines, including EA. hy926, A549, U937, RKO, SW620, HCT-116 and HT-29, and animal cell lines, including CT26.CL25 and sp2/0-Ag14, were investigated in this study. Mixed frozen tissue samples of the heart, lung, liver, spleen and kidney obtained from fatal EEHV1A- or EEHV4-infected cases were homogenized and used for cell inoculation. At 6, 24, 48 and 72 h post infection (hpi), EEHV-inoculated cells were observed for cytopathic effects (CPEs) or were assessed for EEHV infection by immunoperoxidase monolayer assay (IPMA) or quantitative PCR. The results were then compared to those of the mock-infected controls. Replication of EEHV in the tested cells was further determined by immunohistochemistry of cell pellets using anti-EEHV DNA polymerase antibodies or re-inoculated cells with supernatants obtained from passages 2 or 3 of the culture medium. The results reveal that no CPEs were observed in the tested cells, while immunolabeling for EEHV gB was observed in only U937 human myeloid leukemia cells. However, quantitation values of the EEHV terminase gene, as well as those of the EEHV gB or EEHV DNA polymerase proteins in U937 cells, gradually declined from passage 1 to passage 3. The findings of this study indicate that despite poor adaptation in U937 cells, this cell line displays promise and potential to be used for the isolation of EEHV1 and EEHV4 in vitro.

Asian elephants have been an important part of wildlife ecotourism in Thailand for over two decades. Elephants in tourist camps are exposed to a variety of management styles and daily activities that can potentially affect health and welfare. This study investigated relationships between a novel welfare biomarker, immunoglobulin A (IgA), and daily camp activities, and compared results to glucocorticoid (GC) measures. Often no-riding camps are portrayed as providing better welfare than camps that offer riding. Therefore, we predicted that elephants at no-riding camps would have lower GC and higher IgA concentrations, and a low GC/IgA ratio. Forty-four female elephants from six elephant camps were divided into three groups based on riding activities: saddle-riding, bareback-riding, and no-riding. Fecal and salivary samples were collected monthly for 1 year along with evaluations of body condition, foot health, and wounding. Camp environment and management varied among camps, although the major difference was in riding activities. Concentrations of GCs and IgA varied among the working groups, but not always consistently between sample matrices. Overall fecal glucocorticoid metabolite concentrations were lowest in the saddle-riding group. Only in one bareback-riding camp did the elephants exhibit a potentially positive welfare response with a low GC/IgA ratio over time. Other results varied between the two biomarkers, with considerable variability across camps, suggesting there is more to good welfare than whether elephants participate in riding or not. Several other human-induced stressors, like chaining, ankus use, and limited social opportunities are likely to be impacting well-being and should be considered to ensure management practices meet physical and psychological welfare needs.

Thailand is the epicenter of elephant tourism and visiting an elephant camp is a popular activity according to the Tourist Authority of Thailand. However, the welfare of these elephants has been questioned by animal activist groups, international tour operators, and the public. Conclusions that the vast majority of captive elephants are abused often are based on anecdotal evidence and not solid science. So, it is difficult to tease apart emotion, opinion, and fact with regard to what practices are good or bad for elephant welfare. The aim of this paper was to: (1) describe the unique status of captive elephants in Thailand and associated regulations, (2) summarize current issues and challenges facing elephant tourism, (3) review studies conducted on welfare of tourist elephants in Thailand, and (4) offer recommendations for how elephants can be properly cared for under captive conditions in tourist camps. We conclude there are many ways to manage these elephants, and that not all tourist activities are bad for welfare. However, it is essential they be managed in a way that meets physical, physiological and psychological needs, and that management decisions are based on objective data.

This review paper is a synthesis of results from multiple studies that we have conducted over the past several years using similar methodologies to identify factors related to welfare of captive populations of elephants in North American zoos and Thailand tourist camps. Using multiple conservation physiology tools, we found that, despite vastly disparate management systems, there are commonalities in how environmental and husbandry factors affect physical and physiological outcomes. Elephants appear to have better welfare, based on fecal glucocorticoid metabolite (FGM) analyses, when housed under conditions that provide a more enriched, stimulating, and less restrictive environment. We also found it is essential to balance diet and exercise for good body condition and metabolic function. In Thailand, use of tools to control elephants, such as the ankus (i.e., guide, hook) and chains, did not equate to poor welfare per se, nor did riding; however, improper uses were associated with higher wound scores and FGM concentrations. Foot health was good overall in both regions, with cracks being the most common problem, and better foot scores were found in elephants kept on softer substrates. Based on these findings, science-based guidelines are being developed in Thailand, while in North America, changes are being incorporated into elephant standards and husbandry resource guides. Management across venues can be improved by encouraging elephant exploration and exercise, establishing socially compatibility groups, ensuring proper use of tools, and providing balanced diets. We contend there is no “one-size-fits-all” management strategy to guarantee good welfare for elephants, but there are essential needs that must be met regardless of where or how they are managed. Future studies are needed to find ways to better socialize elephants; determine how temperament affects coping styles and resilience; study the importance of good handler-elephant relationships; identify more ways for elephants to engage with the environment; and assess the effect of life history on subsequent physiological and psychological well-being.

Salivary immunoglobulin A (sIgA) has been proposed as a potential indicator of welfare for various species, including Asian elephants, and may be related to adrenal cortisol responses. This study aimed to distinguish circadian rhythm effects on sIgA in male and female Asian elephants and compare patterns to those of salivary cortisol, information that could potentially have welfare implications. Subjects were captive elephants at an elephant camp in Chiang Mai province, Thailand (n = 5 males, 5 females). Salivette® kits were used to collect saliva from each elephant every 4 h from 06:00 to 22:00 h for 3 consecutive days (n = 15 samples/elephant). Enzyme immunoassays were used to quantify concentrations of IgA and cortisol in unextracted saliva. Circadian rhythm patterns were determined using a generalized least-squares method. Both sIgA and cortisol followed a circadian rhythm, although the patterns differed. sIgA displayed a daily quartic trend, whereas cortisol concentrations demonstrated a decreasing linear trend in concentrations throughout the day. There was no clear relationship between patterns of sIgA and salivary cortisol, implying that mechanisms of control and secretion differ. Results demonstrate for the first time that circadian rhythms affect sIgA, and concentrations follow a daily quartic pattern in Asian elephants, so standardizing time of collection is necessary.

Background Elephant camps are among the most attractive destinations in Thailand for tourists from many countries. A wide range of management strategies are used by these camps, which can have varied impacts on health and welfare of elephants. Methods This study surveyed 33 camps with 627 elephants in northern Thailand to quantify the types of management practices and work activities experienced by captive elephants. The survey consisted of an interview with camp owners, and direct observations of camp operations. Results Data revealed considerable variation in elephant demographics, work activities, elephant care (i.e., housing, restraint, nutrition, health care, and breeding), and mahout management among the camps. In general, older camps (those in existence for >16 years) were involved in more intensive activities, like riding with saddles and shows. By contrast, newer camps provided more one-on-one activities for tourists and elephants, and emphasized more intimate, relaxing experiences (e.g., feeding, bathing, walking) than entertainment. A demographic shift also was observed, with elephants 20 years of age and younger having a sex ratio closer to 1:1 compared to elephants in older age categories (1:4.1–1:9.8). Discussion Shifts in elephant management to less intensive activities were observed, which could have positive implications for elephant welfare. The shifting sex ratio suggests successful captive breeding is resulting in the birth of more males, which could present new welfare challenges in the future, because bulls can be more difficult to manage and socialize, and are more likely to be kept isolated during musth. Ultimately, the goal is to understand how camp activities affect welfare, and to develop science-based guidelines and standards to aid in the management of both male and female elephants used in tourism.

Background Elephants are the largest and heaviest living terrestrial animals, but information on their histology is still lacking. This study provides a unique insight into the elephant’s organs and also provides a comparison between juvenile Asian elephants and adult Asian elephants or other species. Here we report on the histological structure of 24 organs, including the skin, brain (cerebrum, cerebellar hemisphere, vermis, thalamus, midbrain), spinal cord, sciatic nerve, striated skeletal muscle, cardiac muscle, bone (flat bone and long bone), cartilage (hyaline cartilage and fibrocartilage), heart (right atrium, right ventricle), blood vessels (aorta, pulmonary artery and caudal vena cava), trunk, trachea, lung, tongue, esophagus, stomach, small intestine (duodenum, jejunum, ileum), large intestine (cecum, colon, rectum), liver and pancreas, kidney, ovary, uterus (body and horn) and spleen of two juvenile Asian elephants. Methods Tissue sections were stained with Harris’s hematoxylin and eosin Y. Results While almost all structures were similar to those of other species or adult elephants, some structures were different from other mammalian species, such as: plexiform bone was found in flat bone only; a thin trachealismuscle was observed in the trachea; and no serous or mucinous glands were found in the submucosa of the trachea. Discussion Histological information from various organs can serve as an important foundation of basal data for future microanatomical studies, and help in the diagnosis and pathogenesis in sick elephants or those with an unknown cause of death.

Studies in western zoo elephants have found relationships between body condition and physiological function, and identified mitigating management strategies to optimize health and welfare. A similar methodological approach was used in this study, which evaluated a body condition score (BCS; 1 = thinnest, 5 = fattest) every other month and fecal glucocorticoid metabolite (FGM) concentrations twice monthly in 33 tourist camp elephants in Thailand for a 1-year period to assess seasonal variations, and determine how lipid profiles [total cholesterol (TC), low density lipoproteins (LDL), high density lipoproteins (HDL), triglycerides (TG)] and metabolic parameters [insulin, glucose, fructosamine, glucose to insulin ratio (G:I)] related to measures of body condition and adrenal function. The most prevalent BCS was 3-3.5 (60.6%), with 27.3% at BCS = 4 (overweight) and 12.1% at BCS = 4.5-5 (very overweight); no elephants had a BCS

Background Primary cultures from Asian elephants (Elephas maximus) allow scientists to obtain representative cells that have conserved most of their original characteristics, function, physiology and biochemistry. This technique has thus gained significant importance as a foundation for further cellular, cell biology and molecular research. Therefore, the aim of this study was to describe conditions for the successful establishment of primary adult fibroblasts from Asian elephant carcasses. Methods Ear tissue sample collection from Asian elephant carcasses and our recommendations are given. We describe here a simple modified protocol for successful isolation and maintenance of primary adult fibroblasts from elephant ear skin. Ear samples from each individual (five 3 × 3 cm2 pieces) were brought to the laboratory within 3 h after collection, kept in transportation medium at 0–4 °C. The ear tissues were prepared by a combination of 10% collagenase type II digestion procedure together with a simple explant procedure. Primary fibroblasts were cultured at 37 °C in Dulbecco’s modified Eagle’s medium (DMEM) with 20% fetal calf serum (FCS) in a humidified atmosphere containing 5% CO2. After the third passage, fibroblasts were routinely trypsinized with 0.25% trypsin/EDTA and cultured in DMEM with 10% FCS at 37 °C and 5% CO2. Traditional cell counting method was used to measure cell viability and growth curve. Long-term storage of cells used freezing medium consisting of 40% FCS (v/v). Results We explored the most suitable conditions during sample collection (post-mortem storage time and sample storage temperature), which is the most important step in determining primary outgrowth. Our study successfully established and cultured primary adult skin fibroblasts obtained from post-mortem E. maximus ear skin tissues from six carcasses, with a success rate of around 83.3%. Outgrowth could be seen 4–12 days after explantation, and epithelial-like cells were found after 4–7 days of culture, while fibroblasts appeared at around day 7–10. The fibroblasts had viability and post-freezing recovery rates of around 97.3 ± 4.3% and 95.5 ± 7.3%, respectively, and doubling time was about 25 h (passage 6). Discussion To our knowledge, this report is the first to describe primary cell cultures derived from adult Asian elephant skin. Future studies should benefit from the information and useful suggestions herein, which may be used as a standard method for establishing primary skin fibroblast cultures in future experiments.

Mineralized tissues accumulate elements that play crucial roles in animal health. Although elemental content of bone, blood and teeth of human and some animal species have been characterized, data for many others are lacking, as well as species comparisons. Here we describe the distribution of elements in horn (Bovidae), antler (Cervidae), teeth and bone (humerus) across a number of species determined by handheld X-ray fluorescence (XRF) to better understand differences and potential biological relevance. A difference in elemental profiles between horns and antlers was observed, possibly due to the outer layer of horns being comprised of keratin, whereas antlers are true bone. Species differences in tissue elemental content may be intrinsic, but also related to feeding habits that contribute to mineral accumulation, particularly for toxic heavy metals. One significant finding was a higher level of iron (Fe) in the humerus bone of elephants compared to other species. This may be an adaptation of the hematopoietic system by distributing Fe throughout the bone rather than the marrow, as elephant humerus lacks a marrow cavity. We also conducted discriminant analysis and found XRF was capable of distinguishing samples from different species, with humerus bone being the best source for species discrimination. For example, we found a 79.2% correct prediction and success rate of 80% for classification between human and non-human humerus bone. These findings show that handheld XRF can serve as an effective tool for the biological study of elemental composition in mineralized tissue samples and may have a forensic application.

Chinese goral (Naemorhedus griseus) are a threatened species in Thailand and the focus of captive breeding for possible reintroduction. However, little is known of their biology or what factors in the captive environment affect welfare. Our objective was to determine the impact of gender, season, and management on goral adrenal activity. We hypothesized that differences in fecal glucocorticoid concentrations would be related to animal density. Fecal samples were collected 3 days/week for 1 year from 63 individuals (n = 32 males, 31 females) at two facilities that house the majority of goral in Thailand: Omkoi Wildlife Sanctuary (Omkoi), an off-exhibit breeding center that houses goral in individual pens (16 pens; n = 8 males, 8 females) and in small family groups (8 pens; n = 8 males, 8 females); and the Chiang Mai Night Safari (NS), a zoo that maintains 31 goral (n = 17 males, 14 females) in one large pen. Glucocorticoid metabolite concentrations were higher in male than female goral at Omkoi throughout the year, and there was a seasonal effect on adrenal activity (p