SERUM VITAMIN D AND SELECTED BIOMARKERS OF CALCIUM HOMEOSTASIS IN ASIAN ELEPHANTS (ELEPHAS MAXIMUS) MANAGED AT A LOW LATITUDE.

An understanding of species-specific vitamin D metabolism and its role in calcium homeostasis is essential for correct diet formulation and development of husbandry protocols for managed nondomestic species. This study documented serum vitamin D metabolites and other analytes involved in calcium homeostasis in Asian elephants (Elephas maximus) managed at a latitude similar to their wild natural habitat. Serum values for 33 elephants managed at a low latitude were measured in the peak of summer, revealing low vitamin D2 (25(OH)D2 2.3 ± 0.6 ng/ ml and 24,25(OH)2D2 2.17 ± 0.52 ng/ml) and nondetectable vitamin D3. Serum minerals (calcium, phosphorus, magnesium), ionized calcium, and parathyroid hormone were within normal reported ranges. In comparison with previously reported values in elephants managed at a high latitude, 25(OH)D2 (P < 0.001), 24,25(OH)2D2 (P = 0.001), and magnesium (P = 0.013) were significantly lower, and parathyroid hormone was significantly higher (P < 0.001). The lack of D3 production during ample sun exposure at a low latitude suggests that Asian elephants are incapable of cutaneous photobiosynthesis of vitamin D, and that low serum D2 is normal for this species.

Penciclovir pharmacokinetics after oral and rectal administration of famciclovir in African elephants (Loxodonta africana) shows that effective concentrations can be achieved from rectal administration, despite lower absorption.

OBJECTIVE: To evaluate the pharmacokinetics of famciclovir and its metabolite penciclovir following a single dose administered orally and rectally in African elephants (Loxodonta africana). ANIMALS: 15 African elephants (6 males and 9 females) of various ages. METHODS: Famciclovir (15 mg/kg) was administered orally or per rectum once, with at least a three-week washout period between administrations. Blood was collected at 13 different timepoints per administration for 6 elephants, occurring between February and March 2020. An additional 9 elephants were sampled at variable timepoints per administration utilizing a sparse sampling design between July 2020 and January 2021. Plasma famciclovir and penciclovir levels were measured via HPLC and fluorescence detection. Pharmacokinetic analysis was completed in the summer of 2021 using noncompartmental analysis and nonlinear mixed-effects modeling. RESULTS: Famciclovir was not detected in any sample, suggesting complete metabolism. Key pharmacokinetic parameters for penciclovir following oral administration were time to maximum concentration (tmax; 2.12 hours), area under the concentration-versus-time curve (AUC; 33.93 µg·h/mL), maximum observed concentration (Cmax; 3.73 µg/mL), and absorption half-life (t1/2; 0.65 hours). Following rectal administration, the values were: tmax, 0.65 hours; AUC, 15.62 µg·h/mL; Cmax, 2.52 µg/mL; and absorption t1/2, 0.13 hours. CONCLUSIONS: Famciclovir was rapidly metabolized to penciclovir. Oral administration resulted in slower absorption but higher maximum plasma concentration and higher AUC compared to rectal administration. CLINICAL RELEVANCE: African elephants administered famciclovir via oral and rectal routes resulted in measurable serum penciclovir, and these findings may be utilized by clinicians treating viral infections in this species.

EVALUATION OF PLASMA CARDIAC TROPONIN I IN ASIAN ELEPHANTS (ELEPHAS MAXIMUS) USING TWO CLINICAL ANALYZERS.

Cardiac troponin I (cTnI) is specific to myocardial tissue, highly conserved across taxa, and a reliable indicator of myocardial disease in human and veterinary medicine. Biomarkers, like cTnI, may be useful for cardiac evaluation of elephants because the application of other modalities is complicated by the size of the animal. The goal of this study was to establish observed ranges for plasma cTnI in Asian elephants (Elephas maximus) measured by two point-of-care analyzers. Blood was collected from captive juvenile (≤15 yr; n = 9), adult (16-50 yr; n = 42), and geriatric (>50 yr; n = 16) elephants. Following centrifugation, heparinized plasma was stored at 5°C prior to and in between analyses on iSTAT (Abbott Point of Care Inc, Princeton, NJ 08540, USA) and HUBI-QUANpro (Humiasis Co, Ltd, Anyang-si 14042, South Korea) analyzers. With the exception of two results, plasma concentrations of cTnI were below the limit of quantification (LOQ < 0.05 ng/ml) for the HUBI-QUANpro (n = 64), which prohibited comparison between the two analyzers. Observed ranges were determined for plasma cTnI concentrations reported by the iSTAT for the entire population sampled (n = 58; mean 0.011 ng/ml; SD ± 0.013 ng/ml; range 0.00-0.07 ng/ml; 95% CI 0.008-0.015 ng/ml; median 0.01 ng/ml) and with outliers excluded (n = 50; mean 0.007 ng/ml; SD ± 0.007 ng/ml; range 0.00-0.02 ng/ml; 95% CI 0.005-0.009 ng/ml; median 0.01 ng/ml). No significant differences were observed between age classes (P = 0.70) or sexes (P = 0.34). Higher cTnI concentrations were significantly correlated with increasing age (Pearson's r = 0.426; P = 0.002). Future studies are warranted to investigate the diagnostic potential of plasma cTnI in Asian elephants.

The importance of manual white blood cell differential counts and platelet estimates in elephant hematology: blood film review is essential.

Unique features of elephant hematology are known challenges in analytical methodology like two types of monocytes typical for members of the Order Afrotheria and platelet counts of the comparatively small elephant platelet. To investigate WBC differential and platelet data generated by an impedance-based hematology analyzer without availability of validated species-specific software for recognition of elephant WBCs and platelets, compared to manual blood film review. Blood samples preserved in ethylenediaminetetraacetic acid (EDTA) of 50 elephants (n = 35 Elephas maximus and n = 15 Loxodonta africana) were used. A Mann-Whitney test for independent samples was used to compare parameters between methods and agreement was tested using Bland-Altman bias plots. All hematological variables, including absolute numbers of heterophils, lymphocytes, monocytes, eosinophils, basophils, and platelets, were significantly different (p < 0.0001) between both methods of analysis, and there was no agreement using Bland-Altman bias plots. Manual review consistently produced higher heterophil and monocyte counts as well as platelet estimates, while the automated analyzer produced higher lymphocyte, eosinophil, and basophil counts. The hematology analyzer did not properly differentiate elephant lymphocytes and monocytes, and did not accurately count elephant platelets. These findings emphasize the importance of manual blood film review as part of elephant complete blood counts in both clinical and research settings and as a basis for the development of hematological reference intervals.

Effects of physiological changes and social life events on adrenal glucocorticoid activity in female zoo-housed Asian elephants (Elephas maximus).

Ensuring good health and welfare is an increasingly important consideration for conservation of endangered species and includes breeding of individuals managed under human care. Understanding how factors in the captive environment affect individual animal wellbeing can be aided by long-term monitoring of biological functioning. This study involved longitudinal assessments (4 to 28 years) of reproductive and adrenal hormones in zoo-housed female Asian elephants (Elephas maximus) (age range 4 to ~71 years) to elucidate patterns in adrenal glucocorticoid (GC) activity in association with reproductive and demographic factors, and examine individual response to major social changes. Concentrations of serum and urinary cortisol covaried more consistently with physiological changes (ovarian cycle phase, puberty, pregnancy, lactational anestrus, and age) than with social life events (births, deaths, and facility transfers). Cortisol fluctuated across the ovarian cycle with mean concentrations being higher in the follicular than in the luteal phase, and concentrations were highest in lactational anestrous compared to all other reproductive states. The elephants in this study exhibited substantial individuality in adrenal GC response to major social change, reinforcing the need to assess welfare on an individual basis and to consider factors influencing the impact of perceived stressors, such as social relationships, social support, temperament, and life history. Outcomes from this study deepen our understanding of Asian elephant physiology and highlight the importance of taking intrinsic patterns of hormone secretion into account when evaluating the impact of external factors. Finally, a better understanding of the impact of social change and resiliency in response to real and perceived stressors allows us to improve social management to enhance welfare in both captive settings and free-ranging environments.

Ovarian cyclicity and prolactin status of African elephants (Loxodonta africana) in North American zoos may be influenced by life experience and individual temperament.

Hyperprolactinemia is an endocrine disorder associated with infertility in many species, including elephants. In a recent survey of zoos accredited by the Association of Zoos and Aquariums (AZA), over half of African elephant females (N = 101) were not cycling normally, 30% of which exhibited hyperprolactinemia. We examined whether life experience and temperament predict ovarian cyclicity and circulating prolactin status in individual African elephant females. We hypothesized that, similar to humans, acyclicity and hyperprolactinemia in elephants will be associated with an apprehensive or fearful, anxious temperament, and an increased number of potentially challenging life events (transfers, deaths and births). Ninety-five adult African elephant females housed at 37 AZA institutions were included in this study. Blood samples were collected twice a month for 1 year to determine ovarian cycle (cycling, n = 44; irregular, n = 13; non-cycling, n = 38) and prolactin (normal, n = 44; low; n = 23; high; n = 28) status. Keeper ratings on a 6-point scale were obtained on 32 temperament traits in 85 of these elephants. We determined that giving birth and being exposed to herd mates entering the facility were positively associated with normal ovarian cycle and prolactin profiles. By contrast, age, serum cortisol, and an increased number of herd mates leaving a facility were negatively associated with both. Contrary to our hypothesis, hyperprolactinemia was associated with a popular and caring temperament rating, whereas consistently low prolactin was associated with a fearful, apprehensive temperament. These findings indicate that pituitary-ovarian function may be impacted by life history (cyclicity) and temperament (prolactin), which should be taken into consideration when making management decisions.

Anthropometric and blood data on a hand-reared captive Asian elephant (Elephas maximus) calf: A retrospective case report.

The anthropometric and blood data of an unsuccessfully hand-reared Asian elephant (Elephas maximus) calf were retrospectively compared with the data for calves raised by their real mothers or allomothers, to identify potential reasons for poor outcomes in the hand-reared case. The hand-reared calf grew normally in terms of body weight and withers height. However, blood biochemical data suggested reduced bone metabolism, low immune status, and malnutrition during its life. Blood bone markers were measured to determine whether a skeletal disorder was present in the Asian elephant calf, which was not clear from the anthropometric data. Monitoring these parameters in hand-reared Asian elephant calves, with the aim of keeping them within the normal range, may increase the success rate of hand-rearing of Asian elephant calves.

SEX DIFFERENCES IN THE REFERENCE INTERVALS OF HEALTH PARAMETERS IN SEMICAPTIVE ASIAN ELEPHANTS (ELEPHAS MAXIMUS) FROM MYANMAR.

The reference intervals of health parameters are valuable tools for veterinarians and conservationists to monitor the health status and viability of endangered species. Natural variation in the health of the long-lived Asian elephant (Elephas maximus) is poorly understood, particularly in relation to differences between males and females. Longitudinal health data were collected from clinical examination, hematology, and serum chemistry analyses over 3 yr from 227 healthy individually marked Asian elephants varying in age and sex. The study population was semicaptive and used in Myanmar's timber industry, but maintained natural feeding and breeding behavior. Body condition score (BCS) and blood pressure were investigated in clinical examinations. Hematological parameters included hematocrit, hemoglobin, total white blood cell count, and differential blood cell counts. Serum chemistry parameters included blood urea nitrogen, creatinine, total protein, albumin, globulins, aspartate aminotransferase, alkaline phosphatase, triglycerides, creatine kinase, glucose, calcium, potassium, sodium, and chloride. To the knowledge of the authors, this is the first description of BCS in an elephant population outside of zoos, and of blood pressure in this species using a novel adaptation of the Intelli Wrap Cuff pressure monitor. Several differences between the sexes were observed, with females generally having higher BCS and triglycerides, and males displaying higher alkaline phosphatase and glucose levels. This study provides important clinical tools that can be used to assess the health status and improve management in this endangered species.

BIOLOGICAL VARIATION OF HEMATOLOGY AND BIOCHEMISTRY PARAMETERS FOR THE ASIAN ELEPHANT (ELEPHAS MAXIMUS), AND APPLICABILITY OF POPULATION-DERIVED REFERENCE INTERVALS.

The aim of this study was to objectively evaluate the biological variation of healthy Asian elephant (Elephas maximus) hematology and biochemistry parameters, therefore enabling evidence-based clinical decision-making to improve patient management. Ten clinically healthy elephants had blood samples collected weekly for 5 wk under standardized conditions. The analytical, between- and within-individual variation, index of individuality, and reference change values were calculated using previously reported methods. Large between-individual variation and small within-individual variation for almost all parameters indicated that individual normal values should be used for interpreting blood results from Asian elephants.

USING IN-HOUSE HEMATOLOGY TO DIRECT DECISION-MAKING IN THE SUCCESSFUL TREATMENT AND MONITORING OF A CLINICAL AND SUBSEQUENTLY SUBCLINICAL CASE OF ELEPHANT ENDOTHELIOTROPIC HERPESVIRUS 1B.

A 3.5-yr-old asymptomatic female Asian elephant (Elephas maximus) with a high load of circulating EEHV1B DNA on qPCR on a routine blood sample, showed progressive depletion of monocytes, lymphocytes, and platelets. Twice daily IV ganciclovir, plasma transfusions, and fluid therapy coincided with a decreasing viral load, which may support potential efficacy of this antiviral drug. An increase in lymphocytes followed initial treatment and preceded the onset of clinical signs. Administration of short-acting glucocorticosteroids for two consecutive days preceded a reduction of lymphocytes, recovery and maturation of monocytes, and gradually decreasing clinical signs, illustrating the potential value of glucocorticosteroids in treatment of clinical EEHV. Three subsequent subclinical episodes with high monocyte and platelet counts did not require intervention. Decision-making was led not just by quantification of viral load and clinical signs, but more specifically by interpretation of the hematological changes using easily accessible, in-house blood smear analysis.

Seroprevalence of Mycobacterium tuberculosis Complex in Free-ranging African Elephants (Loxodonta africana) in Kruger National Park, South Africa.

Tuberculosis (TB) is a pathogenic disease that affects a range of wildlife species, including African elephants (Loxodonta africana). The recent discovery of fatal disease caused by infection with Mycobacterium tuberculosis in a bull elephant in the Kruger National Park (KNP), which is a bovine TB endemic area, emphasizes the importance this disease could have on both wild and captive elephant populations globally. Elephants with culture-confirmed TB have previously been shown to produce strong antibody-responses before the mycobacteria can be isolated. Therefore, we used two serologic assays that detect TB antibodies to retrospectively screen a cohort of 222 free-ranging African elephants sampled between 2004 and 2018 in KNP. The estimated TB seroprevalence for this free-roaming elephant population was between 6% (95% confidence interval [CI], 2-12%) and 9% (95% CI, 6-15%) based on the two tests. Overall, males had a higher TB seroprevalence than females, and adults (≥25 yr) had a higher TB seroprevalence than younger elephants (≤24 yr) on both rapid tests. The relatively high TB seroprevalence that we found highlighted the value of conducting retrospective studies in free-ranging wildlife populations in order to better understand the potential risk of disease.

Oncotic pressure and the effects of water deprivation in healthy captive Asian elephants.

We evaluated the oncotic pressure (plasma colloid osmotic pressure, πc) in a group of healthy, captive Asian elephants (Elephas maximus; n = 21) with a colloid osmometer with a membrane cutoff of >20,000 daltons. The median πc for these elephants was 26.3 mm Hg with an interquartile interval of 25.5-26.8 mm Hg. The mean πc value was 26.0 mm Hg ± SD 1.1. We found moderate correlation between albumin measured by electrophoresis and πc (r = 0.622; p = 0.003). After a 16-h water deprivation test in a subset of elephants (n = 16), a difference in πc was not detected, despite a significant increase in serum total proteins, urea, and osmolality. These results indicate that πc is not a sensitive indicator of hydration status in elephants after a short period of water deprivation. Use of oncotic pressure as a diagnostic tool in diseased Asian elephants warrants further investigation.

Hyperprolactinemic African elephant (Loxodonta africana) females exhibit elevated dopamine, oxytocin and serotonin concentrations compared to normal cycling and noncycling, low prolactin elephants†.

Many zoo elephants do not cycle normally, and for African elephants, it is often associated with hyperprolactinemia. Dopamine agonists successfully treat hyperprolactinemia-induced ovarian dysfunction in women, but not elephants. The objective of this study was to determine how longitudinal dopamine, serotonin, and oxytocin patterns in African elephants are related to ovarian cycle function. We hypothesized that dopamine concentrations are decreased, while oxytocin and serotonin are increased in non-cycling, hyperprolactinemic African elephants. Weekly urine and serum samples were collected for eight consecutive months from 28 female African elephants. Females were categorized as follows: (1) non-cycling with average prolactin concentrations of 15 ng/ml or greater (HIGH; n = 7); (2) non-cycling with average prolactin concentrations below 15 ng/ml (LOW; n = 13); and (3) cycling with normal progestagen and prolactin patterns (CYCLING; n = 8). Both oxytocin and serotonin were elevated in hyperprolactinemic elephants. Thus, we propose that stimulatory factors may play a role in the observed hyperprolactinemia in this species. Interestingly, rather than being reduced as hypothesized, urinary dopamine was elevated in hyperprolactinemic elephants compared to CYCLING and LOW prolactin groups. Despite its apparent lack of regulatory control over prolactin, this new evidence suggests that dopamine synthesis and secretion are not impaired in these elephants, and perhaps are augmented.

Determination of serum bone-specific alkaline phosphatase isoenzyme activity in captive Asian elephants (Elephas maximus) using an agarose gel electrophoresis method.

The bone-specific alkaline phosphatase (ALP) isoenzyme activity was measured in 51 serum samples from four captive Asian elephants (Elephas maximus) using a conventional method with wheat germ lectin precipitation and a commercial agarose gel electrophoresis (AGE) kit; the isoenzymes were designated as bone-specific ALP (BAP) and ALP isoenzyme 3 (ALP3), respectively. This study examined the suitability of the AGE kit for analyzing blood biochemistry in Asian elephants. The serum ALP3 and BAP activities were strongly positively correlated and met the evaluation criteria for agreement using Bland-Altman analysis. The results indicate that the AGE kit can be used to examine the blood biochemistry in Asian elephants instead of the conventional method.

Variation in metabolic factors and gonadal, pituitary, thyroid, and adrenal hormones in association with musth in African and Asian elephant bulls.

Longitudinal analyses of serum testosterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH), prolactin, glucose, insulin, triglycerides, cholesterol, total and free thyroxine (T4), total triiodothyronine (T3), thyroid stimulating hormone (TSH), and cortisol were conducted to investigate pituitary, metabolic, and adrenal changes related to testicular function and musth status in zoo-housed elephant bulls. Blood samples were collected twice a month for 12 months from 14 African and 12 Asian bulls at 17 facilities in North America. Building on previous studies, our results show that musth is associated with increased testosterone, LH, FSH, and cortisol secretion, and a decrease in thyroid hormone (total and free T4) production. In addition, glucose and triglycerides were higher during musth than non-musth periods, indicative of altered sugar and fat metabolism. There were significant differences associated with age for LH, FSH and testosterone, all increasing, whereas the glucose-to-insulin ratio (G:I) decreased with age. A species comparison found African and Asian elephants differed in measures of insulin, prolactin, cholesterol and the G:I. Across all hormones, high inter-individual variability was observed, making it difficult to define a general musth endocrine profile or to assess musth status from single samples. These results highlight the need for facilities hosting bulls to closely and consistently monitor each individual from an early age and throughout musth and non-musth periods to determine the pattern for each male.

Rapid and sensitive detection of elephant endotheliotropic herpesvirus 1 (EEHV1) in blood by loop-mediated isothermal amplification (LAMP).

Elephant endotheliotropic herpesvirus type 1 (EEHV1) is the most important causative agent of an acute fatal hemorrhagic disease in Asian elephants (Elephas maximus). We employed loop-mediated isothermal amplification (LAMP) to develop a rapid and simple detection method for EEHV1 in blood. When used to test 21 clinical samples collected in Japan, the EEHV1 assay correctly identified one positive and 20 negative clinical samples. It was observed that when samples were spiked with synthetic DNA plasmids including EEHV1 polymerase gene, the detection limit of the LAMP assay was 101.2 copies/µl and 100-fold higher than that of conventional PCR. These advantages of the LAMP assay for EEHV1 detection may facilitate better veterinary practices for treating elephants suffering from the acute disease.

EFFECTS OF GONADOTROPIN-RELEASING HORMONE ANTAGONIST DEGARELIX ON MUSTH AND SERUM TESTOSTERONE CONCENTRATIONS IN ASIAN ELEPHANTS ( ELEPHAS MAXIMUS).

Two male Asian elephants (bulls 1 and 2) in musth were subcutaneously injected with a long-acting gonadotropin-releasing hormone (GnRH) antagonist, degarelix acetate (240 µg/kg; total dose of 960 mg). Musth behavior (MB) and temporal gland secretions (TGS) were monitored and serum testosterone concentrations were determined. In bull 1, MB and TGS ceased on day 1 and reappeared 5.5 mo after the treatment (day 0). During the subsequent musth cycle, MB and TGS ceased on day 1 and did not appear for 4 mo. In bull 2, MB and TGS ceased at day 7 after the treatment. Musth behavior and TGS recurred on Day 11 and continued for 1 wk, then disappeared for 8 mo. Serum testosterone concentrations decreased ( P < 0.05) in all occasions from day 0 (29.8 ± 15.8 ng/ml; mean ± SEM) to day 1 (2.2 ± 1.1 ng/ml), suggesting a sudden drop in circulating testosterone in musth elephants after the GnRH-antagonist treatment.

THROMBOELASTOGRAPHY IN THE HEALTHY ASIAN ELEPHANT ( ELEPHAS MAXIMUS): REFERENCE INTERVALS AND EFFECTS OF STORAGE.

Hemorrhagic disease associated with elephant endotheliotropic herpesvirus infection is the most-frequent cause of mortality in captive Asian elephants ( Elephas maximus). Survival relies on intensive monitoring of hemostatic status. Thromboelastography (TEG) utilizes whole blood samples containing all the blood components of hemostasis and is therefore a sensitive indicator of the clinical status in the patient. This study was performed to assess the practicability of TEG in Asian elephants in a zoo environment. Citrated stabilized whole blood samples were obtained from 44 healthy Asian elephants. Kaolin-activated TEG was performed on whole blood at 60 min and 24 hr postsampling (to replicate shipment to an external laboratory) as well as on freeze-thawed plasma samples, 12-14 mo postsampling. Reference intervals were calculated for fresh whole blood and freeze-thawed plasma samples. In the 24-hr analysis, storage artifacts, likely due to cellular degeneration, resulted in a hypercoagulable thromboelastogram and thus reduced sensitivity for detecting coagulopathies. Therefore, delayed analysis of whole blood samples is not recommended.

Metabolic health assessment of zoo elephants: Management factors predicting leptin levels and the glucose-to-insulin ratio and their associations with health parameters.

Screening for metabolic-related health problems can enhance animal welfare, so the purpose of this study was to conduct the first metabolic health assessment of zoo elephants and use epidemiological methods to determine how factors in the captive environment were associated with metabolic hormone concentrations. In addition, we examined relationships between metabolic status and several fitness parameters: foot health, musculoskeletal health, reproductive cyclicity, and body condition. Two blood samples were collected 2 weeks apart from 87 Asian (Elephas maximus) and 105 African (Loxodonta africana) elephants managed by zoos accredited by the Association of Zoos and Aquariums for analysis of serum leptin, insulin, glucose and the glucose-to-insulin ratio (G:I). In females, mean (± SD) leptin concentrations and the G:I were lower (P<0.05) in Asian (3.93 ± 2.21 ng/ml and 110 ± 86 units) compared to African (4.37 ± 2.89 ng/ml and 208 ± 133 units) elephants, respectively. For males, mean leptin and the G:I were 4.99 ± 3.61 ng/ml and 253 ± 181 units for Asian, and 3.72 ± 2.00 ng/ml and 326 ± 231 units for African elephants, respectively, with no differences between species (P>0.05). As mean leptin concentration increased there was an increase in the odds of a female being non-cycling (P = 0.0083). The G:I was associated inversely with body condition (P = 0.0002); as the G:I increased there was a decreased risk of BCS = 4 or 5 as compared to the ideal, or BCS = 3. Neither leptin nor G:I were predictive of foot or musculoskeletal health scores. Factors related to walking and feeding practices were most influential in predicting metabolic status, whereas social and housing factors showed smaller, but significant effects. The metabolic health benefits of walking were detected if the time spent in staff-directed walking was 7 hours or more per week. The most protective feeding practices included implementing a random rather than predictable feeding schedule and limiting the number of methods presentation methods. Results indicate that leptin levels and G:I can be used as predictors of both ovarian cycle function and body condition, and are affected by zoo management in elephants.

ASSESSMENT OF A LANCET-AND-SWAB BLOOD SAMPLING TECHNIQUE FOR SURVEILLANCE OF ELEPHANT ENDOTHELIOTROPIC HERPESVIRUS INFECTION.

Lancing a finger elicits minimal pain in humans and is applied routinely to obtain small volumes of blood for clinical diagnostics. A modified lancet bleeding method and several blood sampling matrices were evaluated in this study for the purpose of routine elephant endotheliotropic herpesvirus (EEHV) surveillance in Asian elephants (Elephas maximus). The procedure enabled weekly sampling from elephants as young as 9 mo of age. The blood sampling matrices were evaluated for their sensitivity measuring ß-actin, tumor necrosis factor α, and/or EEHV-1 by quantitative polymerase chain reaction assays. Foam and flocked swabs produced significantly (P < 0.05) lower quantitation cycles, ie, increased analytical sensitivity, than filter papers, Whatman® FTA cards, or conventional cotton-tipped swabs. The two swab types also demonstrated comparable analytical sensitivity to that of a similar volume of EDTA whole blood for the detection of EEHV-1 DNA. This lancet-and-swab technique proved satisfactory for the detection of EEHV-1 viremia in two Asian elephant calves, and in one instance viremia could be detected 5 days prior to the development of clinical signs. Low blood yield from the lancet application may reduce sensitivity and compromise early detection of viremia. Therefore, standard venipuncture remains the recommended blood sampling method, and training for consistent and regular vein access should continue to be the priority for collections holding elephants. However, if appropriate measures are taken to collect an optimum blood volume, this lancet-and-swab technique offers a suitable alternative for EEHV surveillance in situations where venipuncture may not be practical.