Serum chemistry data in elephants can provide valuable information about organ function (liver, kidney, muscles, intestines, pancreas) and disease conditions (inflammation, infection), hormone levels and toxins. Proteins, AST, ALT, GGT, creatinine, bilirubin, CK, LDH, Ca, P, glucose, Na, Cl, K are part of the comprehensive chemistry panel. To lab diagnosis Blood chemistry Reference values blood Serum/plasma chemistry Refractometry Serum/plasma Chemistry Serum quality Ser um quality Serum chemistry data may help determining the function of certain organ systems. The reliability of the results largely depend on the quality of the sample. Before running any biochemistry tests, the blood should be well clotted, preferably without hemolysis (shown as red colorat ion of the serum). Chemistry data are usually obtained from serum. Some tests can also be run usi ng plasma. Before using plasma, this option should be checked with the test instructions. Test tubes to be used: red-topped serum-tubes with or without a clotting activator. The color of the serum should be light yellow. The figure below shows different serum characteristic: Hemolysis: Red to brown color due to hemolysis (destruction of erythrocytes). This can have a pathological origin or can result from poor sampling/handling; the red-brown color can also be caused by myoglobin after massive muscle damage (rhabdomyolysis) Milky white color due to presence of fat particles in the serum (physiologic shortly after eating or pathological condition) Yellow color due to the presence of bilirubin (liver damage ->icterus). Blueish-red color due to methemoglobin (low venous oxygen saturation). Lipemia Lipemia, which can be a natural occurrence if the elephant has just eaten, can alter several test results. Calcium, phosphorus, total bilirubin or hemoglobin may be falsely elevated. When using a refractometer to measure the total protein remember that the serum must be clear. If not the value may be falsely elevated. Albumin, sodium and potassium may be falsely lower. Lipemia also enhances hemolysis which in turn can affect lab results. But there is a solution: If you refrigerate your sample the lipemic portion will separate and you can use the clear aliquot below the lipid layer. Normal and lipemic elephant serum Reference values Normal serum chemistry values are determined by species, age, gender and reproductive status of the elephant. It is important to have an understanding of the limitations of laboratory values. The term “reference value” is now considered a more appropriate term than “normal value.” Ideally reference values should be established from studies using a minimum of 30 healthy animals and stated selection criteria. Few elephant studies have been conducted to meet this standard. One report describes chemistry results for different genders of Asian elephants used for logging in Myanmar (Santo, 2020). Moreover, many reference ranges are laboratory specific. So it is best to use one laboratory that can help to develop reference ranges for your elephants. Another important point is that a test result that falls outside of the reference is not necessarily clinically significant. Lab values are information that must be used with all the other information that you have when you are faced with a sick elephant. Establishing a baseline during health and performing sequential tests during illness will give the most reliable information. You also want to use a lab that has good quality controls. If you change labs it is advisable to get new healthy baselines. V ery odd results should always be double checked at the same lab. To page top One study in 10 healthy Asian elephants showed that most Asian elephant hematology and biochemistry parameters are highly individual, requiring individual normal values for accurate interpretation (Perrin, 2020). Test result units Another complicating factor when looking at serum chemistry values is that there is a lack of uniformity regarding units and this can be confusing. Most U.S labs use conventional units whereas in Europe they use SI units. There are conversion factors to go from one system to the other but the conversion factor is test specific – so each test has a different conversion factor. You can find SI conversion calculators online, i.g. ht tps://www.amamanualofstyle.com/page/si-conversion-calculator. Liver The liver plays an important role in the following processes: Protein synthesis and degradation (albumin, clotting factors) Carbohydrate and lipid metabolism Breakdown of hemoglobin Storage (fat soluble vitamins) Detoxification Liver enzymes: Aspartate aminotransferase (AST; SGOT) Alkaline phosphatase (ALP) γ-glutamyl transferase (GGT) Bilirubin Bile acids (?) BSP excretion (bromsulphthalein) AST (SGOT) Aspartate aminotransferase, previously known as serum glutamic oxaloacetic transaminase (SGOT) occurs in all cells. Highest levels are in the liver, cardiac muscle, and skeletal muscle. If AST is elevated then you should also look at the creatine kinase (CK) value. If the CK is normal then AST is likely of liver origin. If the CK is elevated or there is obvious muscle trauma then AST may be of muscle origin. Also AST may falsely increase if the sample is hemolyzed. ALP Alkaline phosphatase is also found in all cells with the highest levels in liver, bone, kidney, intestine, and placenta. ALP is not a sensitive indicator of liver disease in the horse and this probably holds true for elephants although research would be needed to confirm this assumption. ALP may increase with disorders such as rickets. ALP levels are normally higher in young animals including elephants. ALP may indicate colostrum absorption. Non-steroidal anti-inflammatory drugs may cause ALP to elevate. GGT Gamma glutamyl transferase (GGT) is liver specific in horses and pigs. Whether it is liver specific in elephants is unknown. GGT is an indicator of cholestasis (the interruption of bile excretion). GGT has been shown to increase in musth bulls and it has been used together with ALP as an indicator to evaluate passive transfer of antibodies to neonates via colostrum. Bilirubin There is not much information about bilirubin in elephants. There are two forms: unconjugated and conjugated bilirubin. The unconjugated is the main form in horses so this may be true for elephants but we don’t know for sure. Unconjugated (indirect) bilirubin is mainly hemoglobin released from old erythrocytes. It is bound to albumin and transported to the liver where it is conjugated. Conjugated (direct) bilirubin is secreted into bile, transferred to the intestine, converted to urobilinogen by intestinal bacteria, and excreted. Elevated bilirubin may be caused by hemolysis; hepatocellular disease that results in reduced functional mass; and intra - or extrahepatic cholestasis or bile duct obstruction. Unconjugated bilirubin predominates in horses with hyperbilirubinemia regardless of etiology whereas in ruminants unconjugated bilirubin is typical. In one report, elevated total bilirubin (4.94 mg/dl) was observed in a female Asian elephant with colic caused by over-zealous feeding of produce. Values for indirect and direct bilirubin were 3.7 mg/dl and 1.2 mg/dl respectively. Tests of hepatic uptake, conjugation and excretion of bilirubin. Diagram from Lattimer, K.S., Mahaffey, E.A., and Prasse, K.W. 2003. Clinical Pathology 4th edition. Blackwell. P.199. Bile acids Bile acids assist with fat digestion. In most species, bile acids are stored in the gall bladder and released into the intestine. However, elephants do not have a gall bladder. There is controversy whether elephants have bile acids. In several cases bile acids were shown to increase in elephants with TB. Bromsulphthalein (BSP) excretion test In the BSP test a dye is injected IV and measured at several points in time post-injection. In the horse, the half-life is 3.5 minutes. Slow clearance time may indicate cholestasis. Although levels have been shown to increase with liver flukes in elephants, it is not a very practical test. Kidney Blo od Urea Nitrogen ( BUN) and creatinine are the main enzymes used to evaluate kidney function in mammals. BUN Elevations in blood urea nitrogen concentration may be due to prerenal causes like inadequate renal perfusion, shock, or diminished blood volume; renal causes like glomerular-nephritis; or postrenal causes like urinary tract obstruction. Blood Urea Nitrogen makes up approximately 75% of the total non-protein nitrogen (NPN) fraction of the blood. BUN is the major end product of protein nitrogen metabolism. It is synthesized by the urea cycle in the liver from ammonia which is produced by amino acid deamination. Urea is excreted mostly by the kidneys, but minimal amounts are also excreted in sweat and degraded in the intestines by bacterial action. Creatinine Creatinine only elevates when disease is severe and there is marked kidney damage. Unfortunately, in elephants these enzymes are not always useful to predict kidney disease. Creatinine may be lower in young elephants; higher in musth bulls. Serum osmolality/urine osmolality. The osmolality reflects the total number of electrolytes in a fluid. To keep the number constant, the kidney excretes the surplus that is present in the liquid fraction of the blood and the osmolality ratio between these two fluids should be <1. If the kidneys fail to maintain this equilibrium, the serum-urine osmolality ratio becomes > 1. Serum dimethyl arginine (sDMA). In domestic animals sDMA is a marker for endothelial dysfunction and early diagnose of renal disease (declining glomerular filtration rate). In one case report it was associated with kidney failure (polycystic kidney disease). To page top These values are from an Asian bull elephant with capture myopathy. The elevations in SGOT and CK are dramatic. ALT (SGPT) did not increase as much but is probably significant. The elevations in BUN and creatinine may have been related to capture myopathy however this bull was chained to a tree and not given access to food or water post-capture so these changes may reflect dehydration. Muscles Muscle enzymes are: Creatine Kinase (CK) Lactate Dehydrogenase (LDH) Aspartate Aminotransferase (AST; SGOT) Alanine Aminotransferase (ALT; SGPT) Evaluating muscle enzymes can help to diagnose muscle pathology. Conditions that may cause elevated muscle enzyme levels include prolonged recumbency, rhabdomyolysis (also called over-exertion, or tying up syndrome), and clostridial myositis. Bacterial endocarditis, and aortic thrombosis are other causes as well as Vit E/Se deficiency and capture myopathy. Working elephants may be at risk for muscle over-exertion disorders especially as the planet heats up. In all of these conditions, muscle cell membranes rupture and enzymes are released into the blood. Conditions that may lead to increased muscle enzymes in serum are: Prolonged recumbency Rhabdomyolysis Clostridial myositis Bacterial endocarditis Aortic thrombosis Vitamin E/ selenium deficiency Capture myopathy Creatine kinase (CK) CK is critical to muscle energy production. Highest levels are in skeletal muscle, cardiac muscle, and brain. Most CK in the serum is of muscle origin and it is the most sensitive indicator of muscle damage. CK rises quickly – within hours. It also returns to normal quickly as long as there is no on-going damage. Levels that remain high indicate an on-going disease process. Hemolysis interferes with the test and causes falsely elevated values. Lactate d ehydrogenase (LDH) LDH is present in all tissues. Muscle, liver, and red blood cells are the usual sources. LDH is not as useful for determining muscle damage because it is not muscle specific. If LDH is elevated and there is no muscle injury then liver problems should be considered. Like CK, LDH will be falsely elevated in the presence of hemolysis. Aspartate aminotransferase (AST) AST was previously known as serum glutamic oxaloacetic transaminase (SGOT). Muscle and liver are the major sources. It is another enzyme to check if muscle damage is suspected. Alanine aminotransferase (ALT) ALT, also known as serum glutamic pyruvic transaminase or SGPT, is considered muscle specific in large domestic animals. Increased levels have been associated with myopathies in a number of species. ALT will increase in recumbent elephants that are down for a long time. Severe muscle damage can occur in case of Capture Myopathy: To page top Calcium Ionized calcium This electrolyte that is involved in many chemical reactions in the body. The active form is ionized calcium (Ca2+) and this parameter gives the best impression of the available calcium. To measure ionized calcium, special heparinized tubes are needed . Ionized calcium should be > 1.5 mmol/L (>5 mg/dL). Low ionized calcium levels are associated with prolonged parturition and dystocia. Total calcium Non-ionized calcium is predominantly conjugated with albumins and expressed as total calcium. Higher levels of total calcium have been reported in very young elephants and in elephants with TB and severe kidney disease. Many pregnant captive elephants develop subclinical hypocalcemia if calcium is not supplemented during pregnancy. When parturition starts, the demand of calcium is high as it essential for uterine contractions that open the cervix and help expel the calf. Calcium is also high in milk. Calcium supplementation during pregnancy is recommended to form a stock supply. However, releasing stock-calcium ions from the bones is a very slow process, so the blood itself should contain enough calc ium conjugated to albumin to supply the uterus and body muscles for these hours of high demand. Total calcium levels lower than 1.5 mmol/l (6 mg/dl) result in recumbency and levels less than 1 mmol/l (4 mg/dl) will result in death. Low total calcium levels are associated with prolonged parturition and dystocia. Total calcium should be 2.8 +/- 0.2 mmol/L (11 +/- 0.8 mg/dL). Other minerals and electrolytes Sodium (Na) Potassium (K) Chloride (Cl) The reference ranges of these electrolyte in elephants are similar to other species and can best be compared with horse values. To page top Proteins Elephants have a higher to tal protein serum level than most mammals; albu min is lower, and globulins are higher. The ratio of albumin to globulin was shown to be lower in one study that compared elephants with and without TB however the number of TB positive elephants in the study was low. Protein electrophoresis separates the albumin and globulin fractions. There are not many reports in elephants. Protein electrophoresis may be useful to monitor inflammatory conditions or problems like TB, herpes, and others. Acute Phase Proteins Acute phase proteins (APPs) are components of the innate immune system that are markers for infection, inflammation, neoplasia, and tissue injury in humans and domestic animals. APPs are produced by the liver in response to cytokines released from leukocytes. They are initially released into serum 24–96 hr following an acute inflammatory stimulus, where they function to promote healing, reestablish homeostasis, and inhibit microbial growth. The main ones are C-reactive protein (CRP) , serum amyloid (SAA) and haptoglobin (HA) . In one study serum samples from 35 healthy Asian elephants were analyzed for these 3 APPS and levels between the values in healthy and unhealthy elephants were compared (Isaza et al., 2014). From this and other studies it seems that SAA may be the most responsive APP in elephants. The APP values in this study in Asian elephants are shown here: C-reactive protein 12.4-122 nmol/l (1.3-12.8 mg/l) Serum amyloid 0-47 mg/l Haptoglobin 0-1.1 mg/l Glucose Glycolysis of glucose in serum or plasma will decrease the glucose level starting shortly after blood collection. Serum or plasma should therefore b e separated from the red blood cells within 30 minutes after collection. Glycolysis can be prevented by using a Na-fluoride tube for blood collection. Significantly lower levels have been noted in one study comparing TB culture positive and negative elephants. In horses, hypoglycemia may be seen with hepatic failure or bacteremia. There is one report of diabetes mellitus in a 50-yr-old Asian elephant (van der Kolk 2011). Hyperadrenocorticism and hyperthyroidism have not been reported in elephants. Increased blood glucose in elephants is likely to be transient. Some of the causes of transient hyperglycemia could be a cute severe colic acute stress, p ost-postprandial, certain drugs (s teroids, x ylazine, p henothiazine) . Hypoglycemia can occur in n eonatal elephants and in cases associated with m alnutrition, m alabsorption, s epsis, e xtreme physical exertion, a dvanced liver disease or n eoplasia. Amylase and lipase Ranges for these enzymes vary tremendously depending on the methodology and the lab so at this point in time they are not very useful tests for elephants. Amylase may increase in case of pancreas, gastro-intestinal, liver and kidney disease Lipase may increase in case of pancreas or kidney disease. One case of pancreatitis in an elephant has been described so far (pers. comm. Susan Mikota, 2023). Lactate Lactate is an important serum parameter to monitor severe, life-threatening conditions in elephants, like septicemia, Disseminated Intravascular Coagulopathy in a EEHV-HD case. Normal values are between 0--0.11 mmol/L (0-1 mg/dL). Values >0.44 mmol/L (4 mg/dL) are indicative for perfusion problems due to DIC. EEHV-HD patients often have lactate value > 0.22 mmol/L (2 mg/dL) (Wiedner, pers. comm. 2022). A comprehensive elephant serum chemistry panel should include: Total protein Albumin BUN Creatinine AST (sGOT) ALT (sGPT) GGT Bilirubin Bile acids (?) CK LDH Na Cl K Ca P Lactate To page top References Steyrer C, Miller M, Hewlett J, Buss P and Hooijberg EH (2021) Reference Intervals for Hematology and Clinical Chemistry for the African Elephant (Loxodonta africana). Front. Vet. Sci. 8:599387 Santos DJ, Franco dos J, John, Nyein UK, and Lummaa VM. 2020. Sex differences in the reference intervals of health parameters in semi-capt ive Asian elephants ( Elephas maximus ) from Myanmar. J.Zoo&Wildl Med 51(1): 25–38 Debbie JG and Clausen B. 1975. Some hematological values of free-ran ging African elephants. Journal of Wildlife Diseases, 11(1):79-82. Isaza R, Wiedner E, Hiser S, Cray C. 2014. Reference intervals for acute phase protein and serum protein electrophoresis values in captive Asian elephants (Elephas maximus ). J. of Vet. Diagn. Invest. 1-6 Perrin, KL, Kristensen AT, Gray C, Nielsen SS, Bertelsen MF, Kjelgaard-Hansen M. 2020. Biological variation of hematology and biochemistry parameters for the Asian elephant (Elephas maximus ), and applicability of population derived reference intervals. Journal of Zoo and Wildlife Medicine, 51(3) : 643-651. Van der Kolk JH, Hoyer MJ, Verstappen FALM, Wolters SABI, Treskes M, Grinwis GCM and Kik MJL (2011). Diabetes mellitus in a 50-year-old captive Asian elephant (Elaphas maximus ) bull, Veterinary Quarterly, 31:2, 99-101. Https://doi.org/10.1080/01652176.2011.585793 To page top

Esophageal spasms are rarely seen in elephants. This case reports describes this condition in an Asian elephant. Water regurgitation indicated the blockage of water, while the animal was unable to swallow any food. A home-made endoscope greatly facilitated the visualization of the esophagus and stomach wall. A standing sedation using detomidine and butorphanol was used during the treatment procedure. No mouth gag was needed to open the mouth. Continue To non-infectious diseases Case report Esophagus spasm Place: Selwo Zoo, Spain Date: 2019 Data provided by: Cecilia Sierra Arqueros, DVM History Species: Asian elephant Accommodation: Zoo Age, gender: 54 years, female For several years this female Asian elephant had episodes of rhythmic contractions in the ventral area of the neck at the entrence to the thorax (video 1). These contractions were only observed during in the cold seasons of the year. At the age of 54 years, she suddenly became unable to swallow her food and water (video 2). Video 1. Rhythmic contractions in the ventral neck area of an Asian elephant Day 1: The elephant tried to drink water. After 10-20 seconds the water came out her mouth again (regurgitation). The regurgitated water was clear and had no abnormal smell (no stomach smell). Appetite: in the morning she ate horse pellets and some roughage, but then she refused bread and apple slices (her favorites!). She tried to eat fresh gras, but after chewing on it, it came out; no smell of stomach contents. Refused to eat anymore. Regular defecation, though the fecal balls became smaller during the following day. The digestion of the fibers had not changed. Water regurgitation Day 2: No change. Oral inspection: 2 small (5 mm Ø) ulcerations on the tongue base, that were not there the day before. The animal did not cooperate as good as she did on the first day. A standing sedation was performed using detomidine and butorphanol. A 2.4 m plastic tube and flexible endoscope could be advanced into the esophagus, reaching the stomach. No mouth opener or gag was used. Gastric fluids were seen, but no obstruction in the esophagus was encountered. Video 2. Regurgitation of water. Endoscope Plastic tube with endoscope advanced into the esophagus. Treatment Rectal fluids Antibiotic + flunixin meglumine + Vit E/selenium Day 3: Standing sedation using detomidine and butorphanol. Treatment: antibiotics, dexamethasone, vitamin B complex, 240 L rectal fluids Day 4: In the morning, the elphant was able to drink water. Nevertheless another standing sedation was performed using detomidine and butorphanol. 3.5 meter tube inserted in esophangus with mini-camera. Antibiotic + dexametasone +vit- sel + complex B + Buscopan Thirty minutes after finishing the procedure, the elephant started to drink and she ate a melon. From that moment on her appetite came back and she did not regurgitate anymore. Differential diagnosis: Esophagus spasm Esophagus constriction: unlikely because this would have been confirmed by endoscopic examination. Botulism . Botulism had occured in the same environment 200 kilometers from this place, resulting in paralysis of the entire body and the death of the 4 affected elephants. Comments: Esophagus spasms in elephants have not been described before. There is only anecdotal evidence of this phenomenon in horses (van der Kolk, pers. comm. 2021). Hypocalcemia is high on the list of the differential diagnoses. Hypocalcemia has also been associated with " hiccups " in Asian elephants, occuring in the cold seasons. The total calcium concentration in this elephant was 2.7 - 2.9 mmol/l while the hematocrite was 48-50% (average normal value 35%), which is an indication that the elephant was dehydrated. The actual total serum calcium concentration in non-dehydrated condition was probably lower: 35/50 x 2.7 = 1.89 mmol/l. Conclusion: Hypocalcemia may have played a major role in this case of esophagus spasm. To page top

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Dystocia is quite common in older elephants and can be associated with old age at first calving, uterine inertia, fetal overweight, intra-uterine death, malposition, malformation, psychological and physical factors or tumors. Retention of the fetus has been described. Dystocia Text by Willem Schaftenaar Obstructed labor, also known as labor dystocia, can be defined as the condition in which the fetus is unable to pass through the birth canal because the passage is physically blocked. As per definition, the birth process has already started, indicated by the drop of blood progesterone to base-line level. This hormonal change should be followed by labor activities within about 3 days. The longer this period takes, the higher the chances are that a dystocia will develop. Monitoring the birth process using transrectal ultrasonographic examination is of great importance, especially in older elephants. Click here to read more about the normal birth process. In elephants, the main causes for dystocia are: Insufficient dilatation of the vaginal vestibulum due to lack of elasticity of the skin that covers the vaginal vestibulum; Malposition Intra-uterine fetal death Relatively too large fetus Uterine inertia (physical and psychological) Too small pelvis of the dam Intra-uterine death Retained fetus T umors/cysts 1. Insufficient dilatation of the vaginal vestibulum due to lack of elasticity of the skin that covers the vaginal vestibulum. This is probably the major cause of dystocia and stillbirth in older elephants getting their first calf. Whether this is caused by insufficient production of estrogens, is unknown. Once the major part of the calf's body has entered the vertical part of the birth canal, the contractions of the uterus have very little (if any) influence on the propulsion of the calf. Gravity force has taken over the forces produced by uterine contractions. In this position, the umbilical cord may have ruptured already or might be compressed, as the calf is literally squeezed into a narrow space. The author has assisted in at least 5 dystocia cases caused by this phenomenon. Too narrow birth canal in an aged nulliparous elephant resulted in the suffocation of the calf in the vertical part of the vaginal vestibulum. Courtesy: Dak Lak Elephant Conservation Center. 2. Malposition is another important cause of dystocia similar to other mammalian species. The malposition may be caused after the calf died in the uterus. This happened in a 29 yr-old Asian elephant when the umbilical cord was twisted twice around the first presenting leg of her calf. As a result, the calf got stuck in a horizontal position, unable to make the necessary longitudinal twist-movement when entering the vaginal vestibulum. Click here to read this case report. 3. Intra-uterine fetal death can be the result of a fatal infection, e.g. cowpox, salmonellosis or any other form of septicemia. Usually there has been a known episode of illness of the dam during her pregnancy. 4. A relatively too large fetus can result in stagnation of the parturition. In several zoos the birth weight of the calves is too high. The body weight of a newborn calf should not exceed 120 kg. Lack of movement and/or an unbalanced diet are the main reasons for overweight of the new born. 5. Uterine inertia can play an important role in stagnation of the birth process. Exhaustion of the uterine muscle is a form of physiological uterine inertia. It can be induced by repeated administration of (high) doses of oxytocin, which has been reported anecdotally several times. Like in horses, elephants may react on environmental disturbances during the birth process by ceasing labor activities. Hypocalcemia can result in a decrease of the uterine muscle contractions. The author has noticed at multiple occasions, that intravenous infusion of 1-2 liter calcium-magnesium borogluconate increased the strength of the contractions. This was confirmed by transrectal ultrasound examination: the position of the uterus in the abdomen was higher than before the infusion. Monitoring blood calcium levels, and more specifically the ionized calcium concentration, should be part of the monitoring of the birth process. The ionized calcium levels should be around 1.25–1.30 mmol/L (van der Kolk, 2008). Finally, excessive body weight and/or lack of exercise of the dam can possibly play a negative role on the uterine fitness. 6. If the p elvis of the dam too small , the fetus may get stuck even before entering the birth canal. The author assisted a unsuccessful fetotomy in an Asian elephant with a relatively small pelvic diameter. The procedure failed, as the fetus's body could not be reached through the episiotomy -opening. 7. Intra-uterine death of the fetus can result in abnormal position of the fetus. Like in other mammalian species, the fetus should actively collaborate with the labor activities of the dam. If a leg or the head is not stretched at the right time, this may cause complete obstruction of the passage. The author has witnessed that rotation along the longitudinal axis of a dead calf (after creating access through episiotomy) resulted in the easy passage of the calf. Figure 1 shows that the calf rotated along its longitudinal axis on its way through the birth canal. Figure 1. Natural rotation along the longitudinal axis facilitated the birth of this healthy Asian elephant calf. 8. Retained fetus Retention of a dead fetus is not uncommon in elephants. It has been reported in African and Asian elephants kept in captivity. No study has been done on the mechanisms that lead to fetal retention. When the blood progesterone level drops to its baseline at the end of pregnancy, parturition should follow within a few days. The length of the period between baseline progesterone values and the actual birth of the calf is not known. However, following the expected physiological pattern of the birth process in other mammalians, birth should take place in about three days, though there are exceptions of calves being born alive after as long as 14 days. If the period of low progesterone levels exceeds 2 weeks, we can call it a prolonged pregnancy or fetal retention. The chances for the calf to survive a prolonged pregnancy are assumed to be low. To date, reported cases of prolonged pregnancies range from 3 to 60 months. Fetal membranes may or may not remain intact. In contrast to other mammalian species, the fetus is not always affected by microbial contamination. Sterile mummification can result in the conservation of the dead fetal carcass, due to the specific anatomy of the birth canal: the total length of the vaginal vestibulum ranges from 1-1,4 m, including a long vertical part and long horizontal part. In one case (Thitaram, 2006) the hind legs were macerated, while the anterior part of the calf (in anterior presentation!) was well conserved. Labour activities occurring around the expected calving date may have been missed. In 1 case (zoo-born 37 yr-old Asian elephant, 6th calf), vague signs of a birth process, including colic-like symptoms were observed 1 day after progesterone blood levels returned to its baseline level (Schaftenaar, 2013). From that moment the birth canal was ultrasonographically monitored. No changes were noted until 13 days later, when ultrasound examination confirmed partial relaxation of the cervix (13.7 mm, figure 2). In the following days the cervix closed again and remained closed until the final birth process started 13 months later. The mummified calf was removed by total fetotomy . The cause of the stagnation of the birth process could be explained by the congenital arthrogryposis of the calf's legs. This dam survived in good health for another 15 years, when it was euthanized because of chronic degenerative joint disease. At necropsy, the wall of the uterus appeared very thick and there was a known fistula (1 mm diameter) in the skin covering the vaginal vestibulum. No other remnants of the fetotomy were observed. Figure 2. Transrectal ultrasonographic examination of the Asian elephant shows the partial relaxation of the cervix uteri, with a 13.7 mm diameter of the cervix canal. Interestingly, the mother of the previous elephant also suffered a fetal retention of her 5th calf at the age of 38 years. This fetus probably died around 18 months of gestation due to severe illness of the dam. A pyometra was suspected based on the white-yellowish vaginal discharge, which at times contained pieces of (assumed) macerated tissue. The dam was humanely euthanized. Post-mortem examination showed massive adhesions between the uterus and the abdominal wall and the spleen, which probably prevented the uterus from contracting and expelling the dead foetus. After its death the fetus remained in the uterus for another 33 months (Pers. comm. Carsten Gröhndal, 2005). Repeated fetal retentions occurred in a herd of African elephants kept in one zoo. Over a period of 11 years, one female retained four female fetuses, sired by the same bull, while another female from the same herd, but sired by a different bull, retained one female fetus. The retention period ranged from 6-19 months. The calf on this photo was 6 months overdue. (Courtesy Colchester Zoo) One episiotomy or vaginal vestibulotomy after fetal retention in a 32 yr-old nulliparous Asian elephant in Thailand has been published (Thitaram, 2006). Four months before the expected birth, loss of vaginal fluid with necrotic pieces of amnion tissue was noticed. Twelve months later, the animal showed clear signs of signs of labor. As the fetus got stuck in the birth canal, a successful episiotomy was performed. Two cases of fetal retention that were successfully handled by episiotomy with manual traction in Nepal were mentioned in a short communication in the journal Gajah. No further details were provided (Mandal, 2013). 9. Tumors/cysts Tumors in the reproductive organs of female elephants are not uncommon. Leiomyomas are the most frequently found tumors. As they are associated with the uterine muscles, large leiomyomas can reduce the power of the uterine contractions. Due to the space they may occupy in the uterus, these tumors usually reduce the chances for a pregnancy and may result in early stage abortion rather than palying a role in retention of the fetus. In nulliparous cows >30 years of age, vaginal cysts can become so extensive that they fill the entire vaginal lumen, which may block the passage of the fetus. A 26 kg lipoma, situated on the peritoneal side of the uterus in a 22 yr-old Asian elephant in a European zoo had restricted the intra-uterine movement of the hind legs of the fetus, which resulted in ankylosis of those legs. As a consequence, the fetus could not be delivered and remained stuck inside the uterus. During the birth process the uterus ruptured and the dam was humanely euthanized (Figure 3 and 4; the lipoma was located outside within the red circle; photos taken by the author). Figure 3 Figure 4 No doubt that there are more conditions that are resulting in dystocia. Much is still unknown and many cases have not been reported. More information has been provided by Hermes et al. (Hermes, 2008). Dealing with dystocia Much depend on the degree of training of the elephant. If the animal is not trained for any medical intervention (e.g. blood collection, rectal examination), any diagnostic procedure or curative treatment must be done under standing sedation . Elephants that are trained for rectal examination should be monitored by ultrasound examination on a daily base once the progesterone level has dropped to baseline value. Ultrasound examination is the most important diagnostic tool to get data of the phase of the birth process. Click here to read more about the parturition process and the options for treatment of dystocia. References Hermes R, Saragusty J, Schaftenaar W, Göritz F, Schmitt DL, Hildebrandt TB. 2008. Obstetrics in elephants. Theriogenology 70 (2008) 131–144. van der Kolk JH, van Leeuwen JPTM, van den Belt AJM, van Schaik RHN, Schaftenaar W. Subclinical hypocalcaemia in captive Asian elephants (Elephas maximus ). Vet Rec 2008;162(15):475–9. Madal RK, and Khadka KK. 2013. Health Status of Captive Asian Elephants in Chitwan National Park, Nepal. 2013. Rabindra Kumar Mandal et al. Gajah 39, page 38. Schaftenaar W. 2013. Delayed postpartum fetotomy in an Asian elephant (Elephas maximus ). Journal of Zoo and Wildlife Medicine 44(1): 130–135, 2013. Thitaram C, Pongsopawijit P, Thongtip N, Angkavanich T, Chansittivej S, Wongkalasin W, Somgird C, Suwankong N, Prachsilpchai W, Suchit K, Clausen B, Boonthong P, Nimtrakul K, Niponkit C, Siritepsongklod S, Roongsri R, Mahasavankul S. 2006. Dystocia following prolonged retention of a dead fetus in an Asian elephant (Elephas maximus ) Theriogenology 66 (2006) 1284–1291. To page top

This chapter describes cases reports submitted by veterinarians worldwide. Case reports Dentistry Back to Top Tusk fracture in 36 months old African elephant Tusk fracture repair in 9 yr-old Asian elephant Tusk fracture repair in 4 yr-old African elephant Tusk (crack) fissure repair in 22 yr-old Asian elephant Tusk sulcus infection in adult Asian elephant T usk sulcus trauma after tusk fracture (2x) Tush loss in female Asian elephant Mandibular fracture Weight loss due to abnormal molar change Supernumerary tusk in an African elephant Dermatology Skin wounds in adult Asian elephant Wound treatment after fetotomy Wound treatment after vaginal vestibulotomy Temporal gland impaction: non-surgical treatment Temporal gland impaction: surgical treatment Back to Top Reproduction Vaginal vestibulotomy (1996) Vaginal vestibulotomy (2014) Fetotomy Back to Top Ophthalmology Bilateral corneal opacity Back to Top Orthopedic problems Partial pad and nail loss in a 54 yrs-old female Asian elephant Back to Top Infectious diseases Salmonellosis in a group of African elephants Salmonellosis in 7 yr-old Asian elephant Colic and Salmonellosis in an adult Asian elephant Salmonella septicemia in an adult Asian elephant Elephant Endotheliotropic Herpes Virus-Hemorrhagic Disease (EEHV-HD) Fasciolasis in a group of African elephants Rabies in an Asian elephant Back to Top Non-infectious diseases Clostridium botulinum in a herd of elephants Back to Top Clostridium perfringens enterotoxemia in a 6 weeks-old African elephant Intoxication Dieffenbacchia intoxication Paraquat intoxication Back to Top Gastro-intestinal problems Esophagus spasm in an adult Asian elephant Esophagus impaction in a 4.5-yrs-old African elephant Hernia mesenterialis and intestinal rupture in an Asian elephant calf (1 year old) Back to Top Miscellaneous Hiccup Asian elephant with suspected hypocalcemia Hiccup Asian elephant with polycystic nephritis Perineal hernia Colic and abdominal surgery Allonursing in an African elephant Back to Top

Leptospirosis in elephants has been described in Asian elephants. This page describes clinical cases and treatment as well as serological evidence of leptospirosis in apparently healthy elephants and the presence of leptospiresin elephant urine. Leptospirosis General information Leptospirosis is a bacterial infection caused by Leptospira spp . It is a common disease in many species of domestic animals. Leptospirosis is zoonotic. The bacteria are dispersed by urine of rodents, especially rats. The organism can survive many weeks in a slightly alkaline moist environment. The route of infection is by percutaneous inoculation of wounds or through mucous membranes. Several serovars have been associated with disease. Usually the liver is the target organ of Leptospira sp . Icterus, anemia, weight loss, (ventral) edema and general malaise are the main clinical signs. Ocular involvement (uveitis and hypopyon) may also occur. In elephants only a few clinical cases of leptospirosis have been reported. In one study in Sri Lanka, urine samples from 13 healthy domesticated elephants were collected on three consecutive days and analyzed for leptospiral DNA (Athapattu, 2019). Four elephants (31%) were confirmed to shed pathogenic leptospires in their urine. DNA sequencing followed by phylogenetic distance measurements revealed that all positive elephants were infected with L. interrogans. This study reveals the possibility that elephants may act as a source of infection for humans and recommends that all domesticated elephants that are in close contact with humans be screened to detect leptospiral shedding. In a study in India, serum samples were collected from 51 captive elephants kept in three different forest ranges (Shivraj, 2009). The samples were subjected to screening for leptospirosis using the microscopic agglutination test (MAT). It was found that out of the 51 samples seven elephant sera (13.72%) showed antibody titers against two serovars of Leptospira interrogans (L. australis and L. canicola) by MAT indicating the presence of infection or due to the past exposure of captive elephants to leptospiral antigens. In Thailand serum from 113 Asian elephants residing in 10 different tourist camps were tested using the microscopic agglutination test against 22 serovars of Leptospira interrogans (Oni, 2007). A seroprevalence of 58 % was found. The prevalent serovars were L. interrogans Sejroe, L. interrogans Tarassovi, L. interrogans Ranarum and L. interrogans Bataviae and L. interrogans Shermani . These results were similar to studies in domestic livestock and stray dogs in the Bangkok district. Because of the potential risk of indirect transmission of Leptospira spp from elephants to humans, 24 environmental samples were collected from an elephant camp area in western Thailand (Chaiwattanarungruengpaisan, 2019). Eighteen samples (75%) were culture-positive for Leptospira spp . The recovered leptospires were mostly derived from water and soil samples from a river and a mud pond, the main areas for recreational activities. The majority of the isolates were classified into the “Pathogens” clade (89%, 16/18) and more than half of the isolates (61%, 11/18) contained species of the “Saprophytes” clade. Notably, two soil isolates from the river beach sampling area were found to contain leptospiral DNA with high similarity to the pathogenic L. interrogans and L. santarosai . The evidence of diverse Leptospira spp ., particularly those belonging to the “Pathogens” clade, suggest that the shared environments of an elephant camp can serve as potential infection sources and may pose a risk to the elephant camp tourists and workers. It was not clear from this study whether the elephants were the source of these Leptospira spp. Leptospirosis in elephants One clinical case described the following clinical signs: chronic weight loss (400 kg) over a 4-month period (Fowler, 2006). Anorexia was profound. Leptospirosis was included in the differential diagnosis when the elephant developed uve itis and hypopyon . Titers for multiple serovars of Leptospira reached 1:12,800. The liver was the organ system infected. Icterus was marked. The sclera and hypopyon were both bright yellow. Total bilirubin reached 160 μmol/L (9.4 mg/dl), and liver enzymes were elevated. Ventral edema became pronounced, accompanied by ulcerating lesions of the vulva and various areas of the skin. The tip of the tail necrosed from vasculitis. Blood urea nitrogen and creatinine levels remained normal throughout the course of the disease, indicating that the urinary tract was not involved. Diagnosis was based on elevated titers for Leptospira serovars plus hypopyon and uveitis. The organism was not isolated nor could antigens be detected by PCR. Two other elephants cohabitating with the ill elephant developed low titers (1:200–400) for Leptospira icterohemorrhagica , but they did not develop clinical disease. Another case was described by Govindarjan (2006): A 16 yr-old Asian elephant bull was off food for one week. He developed icteric mucous membranes and his urine stained yellow (the author of this report did not mention the color in comparison with the color of normal urine, which is yellow as well). An 8-fold increase of antibodies against L. pyrogenes was observed within a 20-day interval. Diagnosis of leptospirosis is based on serological assays (Micro Agglutination Test MAT, ELISA), PCR, dark field microscopy, and silver impregnation staining. Treatment of elephants with clinical leptospirosis consists of the administration of antibiotics (tetracycline, doxycycline). In the case described above by Govindarjan, amoxicillin was given at a dose of 30 g/day IV for 14 days. References Fowler M. 2006. Infectious diseases. In: Biology, Medicine and Surgery of Elephants, Ed. Fowler and Mikota, 2006, 148. O. Oni, K. Sujit, S. Kasemsuwan, T. Sakpuaram, D. U. Pfeiffer. 2007. Seroprevalence of leptospirosis in domesticated Asian elephants (Elephas maximus) in north and west Thailand in 2004 Veterinary Record (2007) 160, 368-371. Sjivraj, M.D. Venkatesha, Sanjeevkumar, B.M.Chandranaik, Rajkumari Sanjukta, P.Giridhar, and C. Renukaprasad (2009). Detection of leptospiral antibodies in thesera of captive elephants. Veterinary World, Vol.2(4): 2009, 133-134. T. P. J. Athapattu, B. R. Fernando, N. Koizumi and C. D.Gamage. 2019. Detection of pathogenic leptospires in the urine of domesticated elephants in Sri Lanka. Acta Tropica Vol. 195 Pages 78-82 Chaiwattanarungruengpaisan, S., Thepapichaikul, W., Paungpin, W., Ketchim, K., Suwanpakdee, S., Thongdee, M., 2020. Potentially Pathogenic Leptospira in the Environment of an Elephant Camp in Thailand. Tropical Medicine and Infectious Disease 5, 183.. doi:10.3390/tropicalmed5040183 Leptospirosis AAZV fact sheet (2018) Leptospirosis EAZWV fact sheet (2003) To infectious diseases