Medicina de espécies zoológicas

O presente relatório refere-se ao estágio curricular realizado no Badoca Safari Park, no âmbito do mestrado integrado em medicina veterinária da Universidade de Évora, na área de medicina de espécies zoológicas Este encontra-se dividido em duas partes, uma relativa às atividades realizadas e casuística, acompanhada de aprofundamento teórico, e outra relacionada com a pesquisa de hemoparasitas em ungulados, através de esfregaços sanguíneos, efetuada no decorrer do estágio. As hemoparasitoses são infeções com potencial zoonótico, transmitidas por vetores, associadas a importantes perdas económicas. O controlo destas afeções tem como obstáculos fatores económicos e sociais, a heterogenicidade de hospedeiros que estes hemoparasitas e respetivos vetores apresentam, assim como o facto das espécies de animais selvagens poderem atuar como hospedeiros reservatórios. A entrada de animais exóticos e saída de autóctones, em zonas endémicas, provoca desequilíbrios na relação entre o parasita e hospedeiro que poderão despoletar episódios de doença; Abstract: Zoo and Wildlife medicine This report refers to the internship held at Badoca Safari Park, as part of the integrated master’s degree in veterinary medicine, at the University of Évora, in the clinic area of wild species and wild animals. This thesis is devided into two parts, one on the activities and cases, accompanied by theoretical studies, and the other related to hemoparasites research on ungulates through blood smears, performed during the intership. The hemoparasitoses are infections with zoonotic potential, transmited by vectors associated with significant economic losses. The control of these affections has as main obstacles the economic and social factors, the heterogeneity of hosts these hemoparasites and respective vectors present, as well as the fact that the wildlife species can act as reservoir hosts. The entry of exotic animals and the disappearance of natives, in endemic areas, causes imbalances in the relationship between parasite and host that can trigger episodes of illness.

SURVEY OF FELINE LEUKEMIA VIRUS AND FELINE CORONAVIRUSES IN CAPTIVE NEOTROPICAL WILD FELIDS FROM SOUTHERN BRAZIL

A total of 57 captive neotropical felids (one Leopardus geoffroyi, 14 Leopardus pardalis, 17 Leopardus wiedii, 22 Leopardus tigrinus, and three Puma yagouaroundi) from the Itaipu Binacional Wildlife Research Center (Refugio Bela Vista, Southern Brazil) were anesthetized for blood collection. Feces samples were available for 44 animals, including one L. geoffroyi, eight L. pardalis, 14 L. wiedii, 20 L. tigrinus, and one P. yagouaroundi. Total DNA and RNA were extracted from blood and feces, respectively, using commercial kits. Blood DNA samples were evaluated by polymerase chain reaction (PCR) for feline leukemia virus (FeLV) proviral DNA, whereas reverse transcriptase-PCR was run on fecal samples for detection of coronavirus RNA. None of the samples were positive for coronaviruses. A male L. pardalis and a female L. tigrinus were positive for FeLV proviral DNA, and identities of PCR products were confirmed by sequencing. This is the first evidence of FeLV proviral DNA in these species in Southern Brazil.

Survey for natural Neospora caninum infection in wild and captive birds

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Intraspecific Variation of Biological Activities in Venoms from Wild and Captive Bothrops jararaca

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Molecular and Serologic Detection of Ehrlichia spp. in Endangered Brazilian Wild Captive Felids

Ehrlichiosis, an emergent tickborne disease that affects both humans and animals, may represent a threat to the survival and preservation of wild felids in Brazil There are few studies of ehrlichiosis in wild felids in Brazil, but Ehrlichia spp are present in domestic cats Antibodies to Ehrlichia canis have been reported in a puma (Puma concolor) In this study we assessed the presence of these hemoparasites in the blood of Brazilian wild captive felids of the 72 animals tested, 5 (7%) were seropositive for the E cams antigen, and L1 (15%) were positive for E emirs DNA sequences We also performed sequence alignment to establish the identity of the parasite species infecting these animals using 16S rRNA and omp-1 genes Sequences based on 16S rRNA were similar to those found in dogs and cats from Thailand, Brazil, China, and Taiwan and with E canis obtained from a single individual (human) in Venezuela Ehrlichia sp sequence from sampled felines based on omp-1. gene was similar to the p28 and p30 multigene family of E canis To our knowledge, this is the first study of molecular detection of Ehrlichia sp in Brazilian wild feline species

Antibodies to Toxoplasma gondii and Neospora caninum in Captive Neotropical and Exotic Wild Canids and Felids

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Immunohistochemical study of pituitary cells in wild and captive Salminus hilarii (Characiformes: Characidae) females during the annual reproductive cycle

Freshwater fish that live exclusively in rivers are at particular risk from fragmentation of the aquatic system, mainly the species that migrate upriver for reproduction. That is the case of Salminus hilarii, an important migratory species currently classified as “almost threatened” in the São Paulo State (Brazil), facing water pollution, dam construction, riparian habitat destruction and environmental changes that are even more serious in this State. Additionally, this species show ovulation dysfunction in captivity. Our studies focused on the identification and distribution of the pituitary cell types in the adenohypophysis of S. hilarii females, including a morphometric analysis that compares pituitary cells from wild and captive broodstocks during the reproductive annual cycle. The morphology of adenohypophysial cells showed differences following the reproductive cycle and the environment. In general, optical density suggested a higher cellular activity during the previtellogenic (growth hormone) and vitellogenic (somatolactin) stages in both environments. Additionally, the nucleus/cell ratio analysis suggested that growth hormone and somatolactin cells were larger in wild than in captive females in most reproductive stages of the annual cycle. In contrast, prolactin hormone showed no variation throughout the reproductive cycle (in both environments). Morphometrical analyses related to reproduction of S. hilarii in different environmental conditions, suggest that somatolactin and growth hormone play an important role in reproduction in teleost and can be responsible for the regulation of associated processes that indirectly affect reproductive status.

STANDARDIZATION OF SOME ELECTROCARDIOGRAPHIC PARAMETERS OF CAPTIVE LEOPARD CATS (LEOPARDUS TIGRINUS)

Thirty-three captive leopard cats, Leopardus tigrinus, were anesthetized with xylazine (1-2 mg/kg) and ketamine (10 mg/kg), and electrocardiograph (ECG) tests were recorded in all leads with 1 cm = 1 mV sensibility and 25 mm/sec speed repeating DII lead at 50 mm/sec speed with the same sensibility. Results expressed by mean and standard deviation were: heart rate (HR) = 107 +/- 17 (bpm); P-wave = 0.048 +/- 0.072 (s) x 0.128 +/- 0.048 (mV); PR interval = 0.101 +/- 0.081 (s); QRS compound = 0.053 +/- 0.012 (s) x 1.446 +/- 0.602 (mV); QT interval = 0.231 +/- 0.028 (s); R-wave (CV(6)LL) = 1.574 +/- 0.527 (mV); R-wave (CV(6)LU) = 1.583 +/- 0.818 (mV); heart rhythm: normal sinus rhythm (15.2%), sinus rhythm with wandering pacemaker (WPM) (60.6%), sinus arrhythmia with WPM (24.2%); electric axis: between +30 degrees and +60 degrees (6.1%), +60 (6.1%), between +60 degrees and +90 degrees (57.6%), +90 degrees (9%), between +90 degrees and +120 degrees (21.2%); ST segment: normal (75.7%), elevation (18.2%), depression (6.1%); T-wave polarity (DII): positive (100%); T-wave (V(10)): absent (6.1%), negative (63.6%), positive (18.2%), and with interference (12.1%). Through ECG data comparison with other species, unique features of Leopardus tigrinus` (leopard cat) ECG parameters were detected. Some of the study animals presented with an R-Wave amplitude that was indicative of left ventricle overload according to patterns for normal domestic cats (Felis cati). Echocardiographic exams revealed normal heart cavities` function and morphology. The aim of this study was to establish some electrocardiographic parameters of captive L. tigrinus.

Distribution of the entodiniomorphid ciliate Troglocorys cava Tokiwa, Modrý, Ito, Pomajbíková, Petrželková, & Imai, , (Entodiniomorphida: Blepharocorythidae) in wild and captive chimpanzees.

Trophozoites of Troglocorys cava were detected in all but one of the wild chimpanzee populations from Rubondo Island (Tanzania), with a prevalence ranging between 20% and 78%. However, the ciliate was absent in all captive groups. Prevalence appeared to increase with the number of sequential samples taken from a particular individual and reached 95.5% in wild individuals sampled at least 4 times.

Evaluation of two minimally invasive techniques for electroencephalogram recording in wild or freely behaving animals.

Insight into the function of sleep may be gained by studying animals in the ecological context in which sleep evolved. Until recently, technological constraints prevented electroencephalogram (EEG) studies of animals sleeping in the wild. However, the recent development of a small recorder (Neurologger 2) that animals can carry on their head permitted the first recordings of sleep in nature. To facilitate sleep studies in the field and to improve the welfare of experimental animals, herein, we test the feasibility of using minimally invasive surface and subcutaneous electrodes to record the EEG in barn owls. The EEG and behaviour of four adult owls in captivity and of four chicks in a nest box in the field were recorded. We scored a 24-h period for each adult bird for wakefulness, slow-wave sleep (SWS), and rapid-eye movement (REM) sleep using 4 s epochs. Although the quality and stability of the EEG signals recorded via subcutaneous electrodes were higher when compared to surface electrodes, the owls' state was readily identifiable using either electrode type. On average, the four adult owls spent 13.28 h awake, 9.64 h in SWS, and 1.05 h in REM sleep. We demonstrate that minimally invasive methods can be used to measure EEG-defined wakefulness, SWS, and REM sleep in owls and probably other animals.

Study of hantavirus infection in captive breed colonies of wild rodents

Wild sigmondontine rodents are known to be the reservoir of several serotypes of New World hantaviruses. The mechanism of viral transmission is by aerosol inhalation of the excreta from infected rodents. Considering that the captive breed colonies of various wild mammals may present a potencial risk for hantaviral transmission, we examined 85 speciemens of Thrichomys spp. (Echimyidae) and 17 speciemens of Nectomys squamipes (Sigmodontinae) from our colony for the presence of hantavirus infections. Blood samples were assayed for the presence of antibodies to Andes nucleocapsid antigen using enzyme-linked immunosorbent assay (ELISA). Additionally, serum samples from workers previously exposed to wild rodents, in the laboratories where the study was conducted, were also tested by ELISA to investigate prevalence of anti-hantavirus IgG antibodies. All blood samples were negative for hantavirus antibodies. Although these results suggest that those rodent's colonies are hantavirus free, the work emphasizes the need for hantavirus serological monitoring in wild colonized rodents and secure handling potentially infected rodents as important biosafety measures.

Myocardial stereology in captive Callithrix kuhlii (Callitrichidae, Primates): healthy animals versus animals affected by wasting marmoset syndrome (WMS)

This study comprised 12 hearts of Wied´s black-tufted-ear marmoset, Callithrix kuhlii (Coimbra-Filho 1985), 6 with Wasting Marmoset Syndrome (WMS) and 6 non-affected. Biometry was performed after death. After necropsy, the hearts were weighed, dissected, fixed in 10% formalin solution (pH 7.2), and processed for optical microscopy at 5µm sections stained with Haematoxylin-Eosin. Quantitative analysis was performed by stereological techniques. The statistical differences between the biometrical and stereological parameters were assessed by the Mann-Whitney test. The morphometric results showed that WMS causes a significant reduction in body and cardiac weights, and also in the volume density of vessels in those animals. Further studies are necessary to understand some of the results shown here.

Diagnosis of Leishmania infantum infection by Polymerase Chain Reaction in wild mammals

Visceral leishmaniasis is a chronic infectious disease caused by Leishmania infantum (synonym: Leishmania chagasi) and transmitted by the sandfly Lutzomyia longipalpis in Brazil. It is an endemic zoonosis in several regions of the country, including Belo Horizonte (State of Minas Gerais). In urban areas, the domestic dog is susceptible and considered the most important animal reservoir. However, L. infantum has been previously diagnosed in other species, including captive primates and canids. This study aimed to evaluate the presence of the agent DNA in captive animals as well as some free ranging animals from the Zoo-Botanical Foundation of Belo Horizonte by Polymerase Chain Reaction. Eighty one blood samples from primates, carnivores, ruminants, edentates, marsupial, and a monogastric herbivore were analyzed. Three primates Alouatta guariba (brown howler monkey), and two canids Speothos venaticus (bush dog) were positive, demonstrating the importance of leishmaniasis control in endemic areas for preservation of wildlife species in captivity.

Comparison of survival rates of captive-reared and wild-bred Mauritius kestrels (Falco punctatus) in a re-introduced population

Re-introduction is a technique widely used in the conservation of threatened bird species. With advances in aviculture the use of captive-produced individuals as the release stock is becoming more commonplace, and ideally, survival of captive-produced, released individuals should be no different from their wild-bred counterparts. During the late 1980s the Critically Endangered Mauritius kestrel (Falco punctatus) was successfully re-introduced into the Bambous mountain range, Mauritius, some 30 years after its local extinction. Between 1987 and 2001 the developing population was closely monitored enabling us to construct re-sighting histories for 88 released and 284 wild-bred kestrels. We used age-structured models in the survival analysis software program MARK to determine if an individual's origin influenced its subsequent survival. Our analysis indicated no compelling evidence for reduced survival among juvenile captive-reared and released individuals, relative to their wild-bred counterparts, across the majority of cohorts and only limited evidence of a cohort-specific effect. This study illustrates that despite the lack of a formal experimental approach it is still feasible to conduct an assessment of re-introduction outcomes and techniques.

Comparison of survival rates of captive-reared and wild-bred Mauritius kestrels (falco punctatus) in a re-introduced population

Re-introduction is a technique widely used in the conservation of threatened bird species. With advances in aviculture the use of captive-produced individuals as the release stock is becoming more commonplace, and ideally, survival of captive-produced, released individuals should be no different from their wild-bred counterparts. During the late 1980s the Critically Endangered Mauritius kestrel (Falco punctatus) was successfully re-introduced into the Bambous mountain range, Mauritius, some 30 years after its local extinction. Between 1987 and 2001 the developing population was closely monitored enabling us to construct re-sighting histories for 88 released and 284 wild-bred kestrels. We used age-structured models in the survival analysis software program MARK to determine if an individual's origin influenced its subsequent survival. Our analysis indicated no compelling evidence for reduced survival among juvenile captive-reared and released individuals, relative to their wild-bred counterparts, across the majority of cohorts and only limited evidence of a cohort-specific effect. This study illustrates that despite the lack of a formal experimental approach it is still feasible to conduct an assessment of re-introduction outcomes and techniques. (C) 2003 Elsevier Ltd. All rights reserved.