First identification of Mycobacterium avium subsp. paratuberculosis in wild ruminants in a zoo in Mexico.

Background and Aim: Paratuberculosis (PTB) is an infectious disease that induces chronic enteritis in ruminants. It is caused by Mycobacterium avium subsp. paratuberculosis (MAP). In this study, we evaluated the presence of MAP using bacteriological, molecular, and anatomopathological studies, based on the clinical suspicion of PTB in a zoo, in an area housing 10 scimitar-horned oryx (Oryx dammah), five giraffes (Giraffa camelopardalis), and three blue wildebeests (Connochaetes taurinus). Materials and Methods: From November 2016 to June 2017, fecal samples were collected from individuals of the three species on four occasions, resulting in a total of 56 fecal samples. In addition, five small intestine samples were collected from the necropsies of three adult scimitar-horned oryx females and two oryx calves. MAP identification was performed through isolation in Herrold's medium with egg yolk, mycobactin, and sodium pyruvate, Ziehl-Neelsen staining, IS900 polymerase chain reaction (IS900 PCR), and anatomopathological examination of intestine samples. Results: Diffuse granulomatous enteritis with abundant acid-fast bacilli was found in two out of five intestine samples from adult scimitar-horned oryx females. MAP was isolated in 7/56 (12.5%) of the fecal samples from four scimitar-horned oryx, one giraffe, and two wildebeest samples. Two out of 5 (40%) samples obtained from scimitar-horned oryx tested positive. IS900 PCR yielded five positive samples (two fecal samples and three small intestine samples). MAP isolates were classified as Type C (Cattle) using type-specific PCR. Conclusion: These results demonstrated the presence of MAP in the area evaluated and indicated the importance of both sampling live animals and conducting postmortem examinations. The use of bacteriological and histopathological diagnostic techniques demonstrated in this study will provide insight into the health status and prevalence of paratuberculosis in wild ruminants under human care.

Brucella melitensis biovar 1 isolation in a captive wildlife population in the United Arab Emirates. First isolation in the scimitar-horned Oryx (Oryx dammah).

In 2013, Brucella melitensis biovar 1 was recovered from the stomach contents of a scimitar-horned Oryx - SHO (Oryx dammah) aborted foetus, and from the articular fluid of a sand gazelle (Gazella marica) in a captive wildlife collection near Abu Dhabi, United Arab Emirates. Other evidence of exposure to the pathogen was collected through serological testing (Rose Bengal test) and B. melitensis-specific PCR of samples from captive wildlife kept in six different enclosures. A Multiple Locus Variable Number of Tandem Repeats (VNTR) Analysis (MLVA) using 15 markers showed that the two strains isolated in animals kept in enclosures, located 1300 m apart from each other, shared an identical genotype. The phylogenetic analysis of MLVA-15 profiles retrieved from the public database suggested that these strains belong to the African clade, clustering regionally in the UAE, Oman and Qatar. This is the first confirmed case of B. melitensis in a SHO, an African antelope extinct in the wild and warrants further investigation.

Paratuberculosis in Captive Scimitar-Horned Oryxes (Oryx dammah).

Paratuberculosis, a chronic disease caused by Mycobacterium avium subsp. paratuberculosis (MAP), in ten scimitar-horned oryxes (SHOs) hosted in an Italian zoological park and originating from a Slovakian flock, was documented by pathology, molecular, cultural, and serological testing. The infection origin in this threatened species was also investigated by genomic analyses. Following the death of six of the 10 SHOs, serial investigations of dead and alive animals were performed. Necropsy, carried out on five out of six animals, identified intestinal thickening and mesenteric lymphadenomegaly in one of the animals. Histopathology (5/6) revealed lepromatous (2/5) and tuberculoid (2/5) intestinal forms or lack of lesions (1/5). Ziehl-Neelsen and immunohistochemistry stains identified two multibacillary, two paucibacillary forms, and one negative case. MAP was identified by quantitative PCR (qPCR) in tissue samples in five out of five SHOs and was microbiologically isolated from two of the three animals whose fresh tissue samples were available. Fecal samples were collected in four of the six dead animals: all four resulted positive to qPCR and in MAP was isolated in three. ELISA identified MAP-specific antibodies in three of the five dead animals whose serum was available. qPCR identified MAP in the freshly deposited feces of two out of the four alive animals. From the feces of these two animals, MAP was microbiologically isolated in one case. All isolates were classified as MAP type C and profiled as INMV2 and MVS27 by molecular analysis. Genomic analysis of a field isolate revealed clusterization with a European clade but was more similar to Italian than East European isolates. Our findings underline that paratuberculosis should always be considered in zoological parks in which endangered species are hosted. Infection can be subclinical, and multiple combined testing techniques may be necessary.

Foot-and-mouth disease outbreaks in captive scimitar-horned oryx (Oryx dammah).

This paper describes three episodes of foot-and-mouth disease (FMD) that were detected during 2013-2015 in scimitar-horned oryx (Oryx dammah) (SHO), a large Sahelo-Saharan antelope extinct in the wild housed in a wild ungulate breeding facility located 50 km east of Abu Dhabi, United Arab Emirates. While no mortality attributable to FMD was noted in the population of nearly 4,000 SHO during two of the three outbreaks, the morbidity varied according to the circulating strains and seroconversion reached a plateau of 78.0% within two weeks and remained at this level for at least nine months. Partial or complete sequencing of the VP1 encoding region demonstrated that the three outbreaks were caused by three different FMDV lineages (O/ME-SA/PanAsia-2, A/ASIA/Iran-05 and O/ME-SA/Ind-2001), consistent with FMD viruses that are circulating elsewhere in the region. These findings demonstrate that SHO are susceptible to FMD and highlight the risks of virus incursion into zoos and captive facilities in the Arabian Peninsula.

Survey of husbandry practices for bovidae in zoos: the importance of parasite management for reintroduction programmes.

Animals from zoological institutions may be used for reintroductions. These individuals are considered healthy, but they are not necessarily free of parasites, despite the minimum husbandry standards required of zoological institutions as described in the European Association of Zoos and Aquaria guidelines. In this sense, parasitism has been identified as the cause of failure, or has added difficulties, in some reintroduction programmes. Here the authors attempt to summarise the risk of parasitism to animals originating from zoological institutions by analysing a questionnaire about parasite prevalence, sampling methods, treatment and control in three ungulates in European zoos. Completed questionnaires were received from 38 institutions (58.5 per cent response rate). Most of the responding institutions (97 per cent) detected the eggs of endoparasites in faeces, but only one reported ectoparasites. Most institutions followed a similar preventive schedule, with ivermectin as the preferred prophylactic treatment for parasites, commonly administered in food every six months. The frequent use of concentrating flotation techniques as the sole method to evaluate the presence of parasite eggs in faecal samples is not recommended because it fails to detect trematode and lung nematode infections, so it would be better to use flotation techniques together with sedimentation procedures or serological and molecular tests. The results suggest that parasite control in zoological institutions can be complicated, indicating the need to implement a specific management schedule for institutions involved in reintroduction projects.

FECAL CORTISOL LEVELS IN SCIMITAR-HORNED ORYX, ORYX DAMMAH, REVEALS DIFFERENCES BETWEEN CAPTIVE ENVIRONMENTS.

Due to the intensive management of the scimitar-horned oryx, Oryx dammah, involving both captivity and reintroductions, understanding the stress associated with environmental situations this endangered species might experience would be particularly helpful. Fecal cortisol levels were measured across seasons, between captive management programs, and among varying reproductive states in animals held at Fossil Rim Wildlife Center (FRWC) and Kansas City Zoo (KCZ). A total of 72 samples were collected from FRWC and 69 samples were collected from KCZ. The herd size and sex ratio changed for both locations in the middle of sampling due to translocations and birth. The herd sizes ranged from 25 to 28 individuals at FRWC and 22 to 24 individuals at KCZ. An ELISA was optimized and utilized to investigate fecal cortisol variances across seasons, captive management programs, and among varying reproductive states. Fecal cortisol levels ranged from 68.9 to 668.7 pg/g throughout this study, but key differences were found in response to reproductive status, environmental conditions, and social status. The highest level of fecal cortisol measured (668.7 pg/g) was in a pregnant female 9 days prior to parturition. During winter months, an increase in fecal cortisol levels occurred in both herds (FRWC 160.3 pg/g to 335.1 pg/g and KCZ 118.8 pg/g to 505.0 pg/g). In addition, when intact males were held together in an enclosure, the dominant males had lower fecal cortisol levels compared with submissive males during three of the four sampling periods. Understanding how these data relate to the physiologic stress response will require further study, but these results can be utilized to help establish expected fecal cortisol ranges in multiple environments and can aid current captive scimitar-horned oryx management programs, as well as future reintroduction efforts.

Fecal parasite identification by microscopy and PCR in scimitar-horned oryx, Oryx dammah, managed at two sites.

The scimitar-horned oryx, Oryx dammah, an endangered species extinct in the wild, is managed in various captive management programs and is the focus of reintroduction efforts. Management variability can contribute to substantial parasite load differences, which can affect deworming programs and potentially transfer parasites to different regions with translocations. Parasite studies in O. dammah are lacking. In this study, we determined fecal egg/oocyst counts of O. dammah in two captive herds, Fossil Rim Wildlife Center (FRWC) and Kansas City Zoo (KCZ). Fecal egg counts (FEC) were performed on O. dammah feces collected seasonally using the modified McMaster method, and microscopy provided additional identification of parasite genera ova and oocysts. To identify parasites to species level, homogenized fecals provided DNA subjected to the polymerase chain reaction (PCR) using genus specific primers. Microscopy and sequencing results indicated the presence of Strongylus (Strongylus vulgaris, Angiostrongylus cantonensis), Trichostrongylus (Haemonchus contortus, Camelostrongylus mentulatus), Trichuris (T. leporis, T. ovis, and T. discolor), Isospora (Isospora gryphoni) and Eimeria (E. zuernii and E. bovis), with Strongylus being the most common. Nematodirus was identified through microscopy at FRWC. Fecal egg counts were significantly higher in (FRWC) than in (KCZ) in all samplings (P = <0.001). No significant difference was seen between parasite load and seasons (P = 0.103), nor site and season (P = 0.51). Both study sites maintained most animals within commonly accepted FEC levels found in domestic livestock. Individuals with high numbers of EPG or OPG were subordinate males, pregnant females, or neonates. Several significant interactions were found between genera of parasites, age, sex, season, and pregnancy status in the FRWC herd. Sampling limitations prevented further analysis of the KCZ herd. Understanding interactions between parasite loads and physiological, environmental, and regional differences can help determine inter-specific transfer of parasites, and establish appropriate anthelmintic programs for O. dammah herds.

Resource partitioning between ungulate populations in arid environments.

Herbivores are major drivers of ecosystem structure, diversity, and function. Resilient ecosystems therefore require viable herbivore populations in a sustainable balance with environmental resource availability. This balance is becoming harder to achieve, with increasingly threatened species reliant on small protected areas in increasingly harsh and unpredictable environments. Arid environments in North Africa exemplify this situation, featuring a biologically distinct species assemblage exposed to extreme and volatile conditions, including habitat loss and climate change-associated threats. Here, we implement an integrated likelihood approach to relate scimitar-horned oryx (Oryx dammah) and dorcas gazelle (Gazella dorcas) density, via dung distance sampling, to habitat, predator, and geographic correlates in Dghoumes National Park, Tunisia. We show how two threatened sympatric ungulates partition resources on the habitat axis, exhibiting nonuniform responses to the same vegetation gradient. Scimitar-horned oryx were positively associated with plant species richness, selecting for vegetated ephemeral watercourses (wadis) dominated by herbaceous cover. Conversely, dorcas gazelle were negatively associated with vegetation density (herbaceous height, litter cover, and herbaceous cover), selecting instead for rocky plains with sparse vegetation. We suggest that adequate plant species richness should be a prerequisite for areas proposed for future ungulate reintroductions in arid and semi-arid environments. This evidence will inform adaptive management of reintroduced ungulates in protected environments, helping managers and planners design sustainable ecosystems and effective conservation programs.

Incidental findings of Cysticercus tenuicollis metacestodes in five oryx species

Objective: To determine the prevalence of Cysticercus tenuicollis (C. tenuicollis) metacestodes in five oryx species kept in Al Ain Zoo animal collection. Methods: This study was based on a retrospective analysis of post-mortem records covering a four year period (July 2010 to July 2014). Results: A total of 213 individual animals were recorded dead during the four year period (July 2010 to July 2014). Out of this, 12 (5.6%) were recorded with C. tenuicollis. More females (8) than males (4) were recorded to have C. tenuicollis, although this was not statistically significant (P = 0.373. 7). Conclusions: This study shows that, Arabian oryx, beisa oryx, fringe-eared oryx, gemsbok and scimitar-horned oryx are susceptible to C. tenuicollis. Based on the epidemiology and the life cycle of this parasite, it is possible that these captive animals ingested the parasite through contaminated feed which could have happened in the pasture land or stray dogs and wild canidae (e.g. fox) visited the zoo contaminating the oryx feed. Stray dogs and wild canidae should be prevented from visiting pasture land and a captive animal facility.

Incidental findings of Cysticercus tenuicollis metacestodes in five oryx species

Objective: To determine the prevalence of Cysticercus tenuicollis (C. tenuicollis) met-acestodes in five oryx species kept in Al Ain Zoo animal collection. Methods: This study was based on a retrospective analysis of post-mortem records covering a four year period (July 2010 to July 2014). Results: A total of 213 individual animals were recorded dead during the four year period (July 2010 to July 2014). Out of this, 12 (5.6%) were recorded with C. tenuicollis. More females (8) than males (4) were recorded to have C. tenuicollis, although this was not statistically significant (P = 0.3737). Conclusions: This study shows that, Arabian oryx, beisa oryx, fringe-eared oryx, gemsbok and scimitar-horned oryx are susceptible to C. tenuicollis. Based on the epidemiology and the life cycle of this parasite, it is possible that these captive animals ingested the parasite through contaminated feed which could have happened in the pasture land or stray dogs and wild canidae (e.g. fox) visited the zoo contaminating the oryx feed. Stray dogs and wild canidae should be prevented from visiting pasture land and a captive animal facility.