Implementing immunocontraception in free-ranging African elephants at Makalali conservancy.

The goal of programmes to provide contraception for elephants should be to formulate an approach that does not require the relocation or immobilisation of the same individual year after year, which would be long-lasting and cause minimal disruption to social and reproductive behaviour. The programmes should be simple to administer, safe and cost-effective, and must meet the objectives defined by managers in the field. An immunocontraceptive programme was initiated in a small free-roaming population of elephants at the Greater Makalali Private Game Reserve in Limpopo Province in 2000 to determine whether the porcine zona pellucida (pZP) vaccine can successfully control population sizes. Further objectives were to determine implementation costs and efficiency through a multi-faceted approach. We have demonstrated that immunocontraception meets the objectives set by managers in the field. Minimal social disruption was observed over the course of treatment, with the mode of delivery (ground or aerial vaccinations) determining the degree of stress within herds and speed of resumption of normal movement patterns. Aerial vaccinations resulted in the least disturbance, with target herds being approachable within a day. In 2005, implementation costs were R880-R1000/elephant/year, inclusive of darts, vaccine, helicopter and veterinary assistance. Irrespective of the source or method of vaccine delivery, a non-pregnant elephant is rendered infertile from 1st vaccine administration. The sooner immunocontraception is implemented, the sooner population growth rates can be controlled. pZP contraception is a realistic alternative management tool, particularly if used as part of a long-term management strategy. Mass-darting from the air eliminates the need for detailed individual histories of each elephant or for employing a person to monitor elephants. Thus, implementation of immunocontraception in larger populations is feasible and practical.

An experimental intratonsilar infection model for bovine tuberculosis in African buffaloes, Syncerus caffer.

An infection model for Mycobacterium bovis in African buffaloes, Syncerus caffer, was developed, using the intratonsilar route of inoculation. Two groups of 11 buffaloes each, aged approximately 18 months, were infected with either 3.2 x 10(2) cfu (low dose) or 3 x 10(4) cfu (high dose) of M. bovis strain isolated from a buffalo. A control group of six buffaloes received saline via the same route. The infection status was monitored in vivo using the comparative intradermal tuberculin test, and in vitro by the modified interferon-gamma assay. All buffaloes were euthanazed 22 weeks post infection and lesion development was assessed by macroscopic examination, culture and histopathology. It was found that the high dose caused macroscopic lesions in nine out of 11 buffaloes. Mycobacterium bovis was isolated from all buffaloes in the high-dose group and from six out of 11 in the low-dose group.

Anaesthesia of nyala (Tragelaphus angasi) with a combination of thiafentanil (A3080), medetomidine and ketamine.

A combination of thiafentanil (A3080), medetomidine hydrochloride (MED) and ketamine hydrochloride (KET) was evaluated in 19 boma-habituated (12 female and 7 males) and 9 free-ranging nyala (7 male and 2 females) (Tragelaphus angasi) to develop a safe and reliable anaesthesia protocol. Wide dosages were used safely during this study with ranges for A3080 of 45 +/- 8 microg/kg with MED of 69 +/- 19 microg/kg and KET of 3.7 +/- 1.0 mg/kg (200 mg/ animal). The dosages developed on boma-habituated nyala proved to be equally effective in 9 adult free-ranging nyala (7 males and 2 females). The optimum dosage for nyala was a combination of A3080 (40-50 microg/kg), MED (60-80 microg/kg) plus 200 mg of KET/animal. The anaesthesia was characterised by a short induction, good muscle relaxation and mild hypoxaemia during monitoring the anaesthesia was rapidly and completely reversed by naltrexone hydrochloride (30 mg/mg of A3080) and atipamezole hydrochloride (5 mg/mg of MED) given intramuscularly. There was no mortality or morbidity associated with this protocol.

Severe hypoxaemia in field-anaesthetised white rhinoceros (Ceratotherium simum) and effects of using tracheal insufflation of oxygen.

White rhinoceros anaesthetised with etorphine and azaperone combination develop adverse physiological changes including hypoxia, hypercapnia, acidosis, tachycardia and hypertension. These changes are more marked in field-anaesthetised rhinoceros. This study was designed to develop a technique to improve safety for field-anaesthetised white rhinoceros by tracheal intubation and oxygen insufflation. Twenty-five free-ranging white rhinoceros were anaesthetised with an etorphine and azaperone combination for translocation or placing microchips in their horns. Once anaesthetised the rhinoceros were monitored prior to crating for transportation or during microchip placement. Physiological measurements included heart and respiratory rate, blood pressure and arterial blood gas samples. Eighteen rhinoceros were intubated using an equine nasogastric tube passed nasally into the trachea and monitored before and after tracheal insufflation with oxygen. Seven rhinoceros were not intubated or insufflated with oxygen and served as controls. All anaesthetised rhinoceros were initially hypoxaemic (percentage arterial haemoglobin oxygen saturation (%O2Sa) = 49% +/- 16 (mean +/- SD) and PaO2 = 4.666 +/- 1.200 kPa (35 +/- 9 mm Hg)), hypercapnic (PaCO2 = 8.265 +/- 1.600 kPa (62 +/- 12 mm Hg)) and acidaemic (pHa = 7.171 +/- 0.073 ). Base excess was -6.7 +/- 3.9 mmol/l, indicating a mild to moderate metabolic acidosis. The rhinoceros were also hypertensive (systolic blood pressure = 21.861 +/- 5.465 kPa (164 +/- 41 mm Hg)) and tachycardic (HR = 107 +/- 31/min). Following nasal tracheal intubation and insufflation, the %O2Sa and PaO2 increased while blood pHa and PaCO2 remained unchanged. Tracheal intubation via the nose is not difficult, and when oxygen is insufflated, the PaO2 and the %O2Sa increases, markedly improving the safety of anaesthesia, but this technique does not correct the hypercapnoea or acidosis. After regaining their feet following reversal of the anaesthesia, the animals' blood gas values return towards normality.

The gamma-interferon test: its usefulness in a bovine tuberculosis survey in African buffaloes (Syncerus caffer) in the Kruger National Park.

A survey to determine the bovine tuberculosis status of buffalo herds north of the Olifants River in the Kruger National Park was conducted, using a new diagnostic approach. Diagnosis of Mycobacterium bovis infection was accomplished using the gamma-interferon assay technique in 608 adult buffaloes out of a total of 29 discreet herds. The animals were immobilized in groups of 10-15, bled, individually marked and then revived and released on site. As soon as test results were available (after 26-36 h), the same buffalo herd was relocated by tracking the frequency of a radio-collar previously fitted to one adult cow per group during the initial operation. Bovine reactors were identified, darted and euthanased from the helicopter. Necropsy and culture findings of all culled buffaloes showed excellent correlation with the results of the ante-mortem gamma-interferon test. The survey revealed that over and above the two positive herds that had been identified during a previous survey carried out in 1996, there were three additional, but previously unidentified, infected herds in the region north of the Olifants River.

Field applications of immunocontraception in African elephants (Loxodonta africana).

The primary aim of the Makalali elephant immunocontraception programme is to test the efficacy of porcine zona pellucida (PZP) vaccine for practical population control of elephants in small, enclosed reserves, with the goal of stabilizing the current growth rate and reducing it to the 5-10% per annum displayed currently in the Kruger National Park. A secondary aim is to test the hypothesis that PZP treatment does not affect patterns of elephant social behaviour. Eighteen sexually mature cows (age > 12 years) were vaccinated in May 2000 using remote darts. Behavioural observations before, during and after vaccination included noting the activity of individual animals every minute for 15 min. No changes in general behaviour patterns have been noted to date although the animals' spatial use of the reserve was erratic during the period of vaccination, indicating irregular or disturbed patterns associated with vaccination. Normality was resumed on completion of the vaccinations. No aggressive or indifferent behaviour related to nursing, calf proximity or bull-cow interactions have been noted. Ten of the females were in various stages of pregnancy when treated. Subsequently, seven of them gave birth to healthy calves and the other three females are expected to calve shortly. It is too early in the study to draw conclusions about stabilization of growth rates.

Anaesthesia of roan antelope (Hippotragus equinus) with a combination of A3080, medetomidine and ketamine.

A dose range was determined for anaesthesia of 20 recently boma-captured roan antelope (Hippotragus equinus) with the synthetic opiate A3080 combined with medetomidine and ketamine. A dose of 10-30 micro/kg A3080 (x = 20+/-8 microg/kg) combined with 5-21 microg/kg medetomidine (x = 13+/-7 microg/kg) plus 0.29-1.11 mg/kg ketamine (x = 0.71+/-0.24 mg/kg) was found to be safe and effective for the field conditions in this study. The anaesthesia produced by this drug combination was predictable and characterised by a short induction time, good muscle relaxation, and acceptable physiological parameters for anaesthesia periods ranging from 49-103 min (x = 64+/-19 min). The wide range (3-4-fold) of doses with acceptable results is also an indication that this drug combination has a wide margin of safety in roan antelope, making it desirable for field use. When 2 dose levels (2-3-fold dif ference) were retrospectively evaluated, no statistical difference was found in induction times, and no observable clinical differences in the anaesthetic episodes were seen. Based on this study, the recommended dose range in roan antelope for this combination is 10-13 microg/kg A3080, 5-6 microg/kg medetomidine and 0.3-0.6 mg/kg ketamine. The anaesthesia produced by this combination was rapidly and completely reversed by i.m. or i.v. injections of naltrexone at 30 times the A3080 dose (x = 0.60+/-0.25 mg/kg) and atipamezole at 3 times the medetomidine dose (x = 38+/-20 microg/kg). No residual effects from ketamine were noted following reversal of A3080 and medetomidine. No mortality was associated with this protocol.

Anaesthesia of gemsbok (Oryx gazella) with a combination of A3080, medetomidine and ketamine.

An effective anaesthesia protocol was developed for adult free-ranging gemsbok (Oryx gazella) using a combination of A3080, medetomidine and ketamine. A short induction time; good muscle relaxation, adequate oxygenation and stable heart rate and respiration rate characterised this anaesthetic regime. Equal doses of A3080 and medetomidine (22-45 microg/kg) plus 200 mg of ketamine were administered to each animal. The anaesthesia was rapidly and completely reversed by intramuscular naltrexone at a dose of X = 0.9 +/- 0.2 mg/kg and atipamezole at a dose X +/- 90 +/- 20 microg/kg. No mortality or morbidity occurred with this protocol.

Use of medetomidine and ketamine for immobilization of free-ranging giraffes.

OBJECTIVE: To develop a dosage correlated with shoulder height (SH) in centimeters for effective immobilization of free-ranging giraffes, using a combination of medetomidine (MED) and ketamine (KET) and reversal with atipamezole (ATP). DESIGN: Prospective study. ANIMALS: 23 free-ranging giraffes. PROCEDURE: The drug combination (MED and KET) was administered by use of a projectile dart. Quality of induction, quality of immobilization, and time to recovery following injection of ATP were evaluated. Physiologic variables measured during immobilization included PaO2, PaCO2, oxygen saturation, end-tidal CO2, blood pH, indirect arterial blood pressure, heart and respiratory rates, and rectal temperature. RESULTS: Sixteen giraffes became recumbent with a dosage (mean +/- SD) of 143 +/- 29 microg of MED and 2.7 +/- 0.6 mg of KET/cm of SH. Initially, giraffes were atactic and progressed to lateral recumbency. Three giraffes required casting with ropes for data collection, with dosages of 166 +/- 5 microg of MED and 3.2 +/- 0.6 mg of KET/cm of SH. Four giraffes required administration of etorphine (n = 2) or were cast with ropes (2) for capture but remained dangerous to personnel once recumbent, precluding data collection. In giraffes successfully immobilized, physiologic monitoring revealed hypoxia and increased respiratory rates. Values for PaCO2, end-tidal CO2, and heart rate remained within reference ranges. All giraffes were hypertensive and had a slight increase in rectal temperature. Atipamezole was administered at 340 +/- 20 microg/cm of SH, resulting in rapid and smooth recoveries. CONCLUSIONS AND CLINICAL RELEVANCE: Medetomidine and KET was an effective immobilizing combination for free-ranging giraffes; however, at the dosages used, it does not induce adequate analgesia for major manipulative procedures. Quality of induction and immobilization were enhanced if the giraffe was calm. Reversal was rapid and complete following injection of ATP.

The rise and fall of tuberculosis in a free-ranging chacma baboon troop in the Kruger National Park.

A single troop of free-ranging chacma baboons (Papio ursinus) was found to be infected with tuberculosis caused by Mycobacterium bovis. It is assumed that some members of the troop originally became infected when feeding on a tuberculous carcass in the veld or on tuberculous material scavenged at a nearby post mortem facility. Subsequently, apparent aerosol transmission took place while sleeping in an unused room. Oral transmission probably also occurred due to continuous contamination of the floor of this room and the common, narrow access (a train bridge crossing the Sabi River) to it with faeces and urine. A macroscopic prevalence of 50 % was found and the disease was noted to progress rapidly in infected baboons. A variety of organs had typical tuberculous lesions, of which the spleen, lungs and mesenteric lymph nodes were consistently, grossly affected. Using Restriction Fragment Length Polymorphism analysis, all but one of the baboon isolates were found to be identical to the most common African buffalo (Syncerus caffer) isolate (genotype 1) in this Park. The opportunistic sleeping facility was made inaccessible to the troop, which was forced to revert to sleeping in trees. A follow-up survey six months after closure, demonstrated that the disease had disappeared from the troop, and that no spillover infection had occurred into neighbouring troops.

Copper poisoning in wild ruminants in the Kruger National Park: geobotanical and environmental investigation.

A geobotanical and environmental investigation was undertaken to investigate the potential for copper poisoning in wild ruminants within the Phalaborwa area in the Kruger National Park and to confirm that environmental copper pollution associated with smelting operations at a nearby mine was the source of copper responsible for the poisoning. The study area selected was divided into high, moderate and low-risk zones and a control area based on initial topsoil copper concentrations and impala (Aepyceros melampus) liver copper concentrations in relation to distance from the copper smelter. Samples collected revealed that topsoil copper concentrations were significantly higher than subsoil copper concentrations at the same sites. There was a significant linear decrease in topsoil to subsoil copper concentration relative to distance from the copper smelter and thus from the high-risk zone to the control area. Copper concentrations of unwashed plant material were significantly higher than washed plant material at the same sites, indicating the deposition of copper on the plant surfaces. Copper deposits in dust fall buckets were significantly higher downwind than upwind from the smelter stack and the presence of atmospheric copper was also confirmed with the aid of low-volume air sampling monitors. The investigation confirmed that the emissions from the copper smelter were sufficient in amount and appropriate in direction to have contributed significantly to the topsoil copper concentrations, unwashed plant copper concentrations and dust fall results.

Sperm abnormalities associated with high copper levels in impala (Aepyceros melampus) in the Kruger National Park, South Africa.

The morphology of spermatozoa may be affected by very low concentrations of pollutants in the environment, and sublethal effects of toxic substances can be detected by studying the sperm of an animal. Sperm were collected from 50 impala (Aepyceros melampus) from a conservation area in South Africa and studied ultramicroscopically. Thirty-two samples were obtained from animals close to a copper mine, while 18 represented animals from an unpolluted area and held in captivity. Liver samples from all these animals were analyzed for their copper content. The findings from 12 animals kept in captivity and fed on a diet to which copper was added correlated with the findings from the field study. The general condition of the animals as expressed by the fat content of the bone marrow from the area exposed to copper contamination was affected detrimentally in comparison to the animals from the uncontaminated area. High percentages of sperm with neck vacuoles were found in all animals with high liver copper levels. A statistically significant correlation is demonstrated between the percentage of impala sperm with vacuoles in the neck region and the copper concentrations in this group of animals.

Copper poisoning in the Kruger National Park: field investigation in wild ruminants.

Chronic copper poisoning was investigated in ruminants within the Phalaborwa area of the Kruger National Park (KNP). Exposure of ruminants to environmental copper pollution resulting form copper smelting operations of a mine in the area was examined by comparing impala faecal copper concentrations in dung heaps and tissue (liver, lung and kidney) copper concentrations of organs collected from impala and buffalo culled within three risk zones (high, moderate and low) of the study area in relation to the distance from the smelter over a period of 4 years. An additional area within the KNP not exposed to the environmental copper pollution from the mine served as control. Tissue copper accumulation was also determined in tracer impala placed in the highest risk zone. The results of this study confirmed the occurrence of chronic copper poisoning in impala and indicated an inverse relationship in extent of impala faecal copper elimination and in tissue copper accumulation in impala and buffalo with distance from the copper smelter. Impala liver copper concentrations were shown to be a reliable indicator of copper accumulation for these ruminants. The presence lung copper concentrations, indicating the exposure to airborne copper were the highest in impala culled in the zone closest to the smelter. Liver copper concentrations above the diagnostic limit of 150 ppm for chronic copper poisoning in domestic sheep were consistently found in impala within the highest risk zone. Clinical pathological measurements suggested that AST activity could possibly be used as an indicator for chronic copper poisoning in impala. It is concluded that, in addition to the environmental and geo-botanical evidence previously reported, the copper smelter of a nearby copper mine is the most likely source of copper pollution responsible for chronic copper poisoning in impala and the occurrence of high copper concentrations in buffalo in the Phalaborwa area of the KNP.

Attempted induction of chronic copper poisoning in boma confined impala.

Induction of chronic copper poisoning in ten boma-confined impala was attempted in a randomized, single dose, parallel designed, titration study using five increasing oral doses, ranging between 125 mg/kg to 1000 mg/kg, of copper oxide needles. Two untreated impala were kept as controls. Impala (n = 1) from each treatment group were culled 52 d and 105 d after treatment and examined for tissue copper accumulation and signs of chronic copper poisoning. Despite the high doses of copper administered to the impala and liver copper concentrations above 150 ppm WM achieved in two animals, no clinical signs related to chronic copper poisoning were observed. Faecal copper concentrations indicated that the major portion of copper oxide particles was excreted in the faeces.

Contraceptive potential of the porcine zona pellucida vaccine in the African elephant (Loxodonta africana).

Immunocontraception has been successful in controlling free-roaming equids; however, what is the potential for the immunocontraceptive control of the African elephant (Loxodonta africana)? The porcine zona pellucida (pZP) glycoproteins share antigenic domains with the African elephant zona pellucida (elZP) glycoproteins, and anti-zona pellucida serum antibodies have been successfully stimulated. To determine the cross-reactivity of the pZP and elZP, immunocytochemistry was evaluated by light and electron microscopy. Specifically, the binding of polyclonal antibodies against total heat-solubilized-porcine zona pellucida to fixed elephant ovary sections was evaluated. The elZP of primary, secondary and tertiary follicles was recognized by the rabbit-anti-pZP serum, but there was no apparent recognition of the primordial follicles. The ability of anti-pZP antibodies to recognize the elZP demonstrates that there is molecular homology between the pZP and elZP glycoproteins. This homology makes the African elephant a candidate for pZP immunocontraception. Three captive elephants were vaccinated with 400 micrograms pZP with a synthetic trehalose dicorynomycolate (S-TDCM) adjuvant. The elephants received 2 boosters of 600 micrograms pZP at 4 wk and 10 m.o. after the primary vaccination. The vaccinated female elephants developed significant (P < 0.05) titers to pZP over prevaccination levels. These levels persisted for 12 to 14 m.o. after the third vaccination. This preliminary evidence shows that the female elephant can develop significant serum antibody levels to pZP. These levels of antibodies are comparable to those required in horses for successful immunocontraception. Thus, porcine zona pellucida immunocontraception might be used to control elephant populations.

Ulcerative pododermatitis in free-ranging African elephant (Loxodonta africana) in the Kruger National Park.

The occurrence of severe lameness in adult African elephant bulls in a shrub Mopane (Colophospermum mopane) ecosystem was investigated. Large ulcers in the soles of at least one front foot were seen in each of the recorded cases. Microscopically, the lesion can be described as a severe, chronic-active, ulcerative, bacterial pododermatitis (complicated by hypersensitivity/septic vasculitis). A variety of bacteria were isolated from these lesions as well as from regional lymph nodes. Streptococcus agalactiae was the most consistent isolate, while Dichelobacter nodosus, the only organism known to be involved with foot disease in domestic ruminants, was isolated from two cases. Contributory factors such as body mass, portal of entry and origin of potential pathogens may have predisposed to the development of the lesions.

An outbreak of encephalomyocarditis-virus infection in free-ranging African elephants in the Kruger National Park.

A cluster of four deaths in late December 1993, marked the onset of an outbreak of disease of African elephants (Loxodonta africana) in the Kruger National Park (KNP) in South Africa, which has an estimated population of 7,500 elephants. Mortalities peaked in January 1994, with 32 deaths, and then declined steadily to reach pre-outbreak levels by September, but sporadic losses continued until November. During the outbreak altogether 64 elephants died, of which 53 (83%) were adult bulls. Archival records revealed that, in addition to the usual losses from known causes such as poaching and intraspecific fighting, sporadic deaths from unexplained causes had, in fact, occurred in widely scattered locations from at least 1987 onwards, and from that time until the perceived outbreak of disease there had been 48 such deaths involving 33 (69%) adult bulls. Carcases had frequently become decomposed or had been scavenged by the time they were found, but seven of eight elephants examined early in 1994 had lesions of cardiac failure suggestive of encephalomyocarditis (EMC)-virus infection, and the virus was isolated from the heart muscles of three fresh carcases. The results of tests for neutralizing antibody on 362 elephant sera collected for unrelated purposes from 1984 onwards and kept frozen, indicated that the virus had been present in the KNP since at least 1987. Antibody prevalences of 62 of 116 (53%) 18 of 139 (13%) and seven of 33 (21%) were found in elephants in three different regions of the KNP in 1993 and 1994. Studies had been conducted on myomorph rodents in the KNP for unrelated purposes since 1984, and trapping attempts were increased during the perceived outbreak of disease in elephants. There was a striking temporal correlation between the occurrence of a population explosion (as evidenced by markedly increased catch rates per trap-night) and a surge in prevalence of antibody to EM virus in rodents, and the occurrence of the outbreak of disease in elephants.