Gastrointestinal parasites in wild and exotic animals from a Zoobotanical Park in Northeast of Brazil

One of the obstacles for ex situ conservation of wild and exotic animals are the diseases that affect them in captivity and, among them, the endoparasitoses, which are very frequent. The objective of this study is to report the occurrence of endoparasites in the fecal samples of animals from the Arruda Câmara Zoobotanical Park, at João Pessoa, Paraiba State, Brazil, as well as to identify significant statistical differences between the percentages of each parasite species found. To this end, 66 fecal samples were obtained from 50 species of animals including mammals, birds, and reptiles. Statistical differences between the percentages of each parasite species were obtained using the binomial test at 5% Research, Society and Development, v. 10, n. 13, e486101321255, 2021 (CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v10i13.21255 2 significance level. A total of 54.5% (36/66) of the fecal samples were positive (p=0.539), among which 80.5% (29/36) presented nematodes, followed by cestodes 19.4% (07/36), protozoans 13.9% (05/36), and mites 16.7% (06/36). This study reports for the first time the association of Balantidium sp. with Tapirus terrestris, Bertiella sp. with Alouatta caraya, Hymenolepis spp., and Aspiculuris spp. as spurious parasites in reptiles; and Entamoeba coli and Eimeria spp. with Iguana iguana in the northeastern part of the country. This study provides knowledge about some of the endoparasites that may occur in zoos in the northeastern region of the country, as well as in expanding the ecological data on wild and exotic animals.


Introduction
Brazil is the fifth largest country in the world with great diversity of animal species. It is estimated that approximately 20% of all animal species live in this region, in different biomes such as the Atlantic Forest, Pampa, Cerrado, Pantanal, Amazon, and the Caatinga. Among the various types of association existing in nature, parasitism is common, and it is well known that at least half of all the animal taxa is parasitic (Instituto Chico Mendes de Conservação da Biodiversidade, 2017; Lima et al., 2017).
One of the obstacles for ex situ conservation of wild and exotic animals are the diseases that affect them in captivity.
Among them, gastrointestinal endoparasitoses are very frequent, and the disease may present itself with or without evident clinical signs. The costs to the host may be negligible, substantial, or even unbearable, depending on the number of parasites, the species, the level of severity of the lesions they inflict, as well as on the vigor and nutritional status of the host. Knowing this, the costs of parasitism may include the loss of resources extracted by the parasite directly from the host and the energy spent by the host to maintain its vital actions. The consequences of these associations are usually linked to the decreased Research, Society andDevelopment, v. 10, n. 13, e486101321255, 2021 (CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v10i13.21255 3 reproductive and survival capacity of the hosts, therefore affecting their mortality and birth rates (Oliveira et al., 2011;Reed et al., 2012;Lima et al., 2017).
Parasitic diseases present a high prevalence in captive wild and exotic animals, since there is a high environmental contamination that result from keeping animals in confined areas. In addition, the stress caused by the captivity may decrease the resistance of these animals to parasitic diseases. In cases in which animals show signs of parasitic disease, these may range from lack of appetite, weight loss, disorders in the gastrointestinal tract, anemia, and hyperthermia, among others (Fagiolini et al., 2010;Oliveira et al., 2011;Reed et al., 2012).
Among the endoparasites that affect zoo animals, there are also many zoonotic species, therefore representing a public health problem, especially for the professionals and keepers who manage these specimens. Many nematodes that can be found in these animals are of medical concern, such as Ancylostoma sp., which can cause cutaneous larva migrans; and Toxocara sp., responsible for visceral larva migrans, ocular larva migrans, and cerebral toxocariasis in humans; both being endoparasites widely reported in carnivores. Further, it is also important to highlight the presence of cestodes such as Hymenolepis spp., Taenia spp., and Bertiella spp., which can be found both in primates and rodents, or even as spurious parasites in reptiles, for example. All of these parasites present medical interest and may cause diarrhea, abdominal pain, irritability, and weight loss (Fagiolini et al., 2010;Oliveira et al., 2011;Reed et al., 2012;Lima et al., 2017).
Thus, these sites should have efficient diagnostic techniques to carry out coproparasitological research in wild and exotic animals in zoos. Among these techniques, the Mini-FLOTAC allows the simultaneous diagnosis of helminth eggs/larvae and oocysts/cysts of protozoans, offering an advantage over other coproparasitological techniques. In addition, Mini-FLOTAC also allows the diagnosis of yeasts, such as Macrorhabdus ornithogaster, in bird feces (Cringoli et al., 2017).
All this knowledge provides new data on the parasites of these host species, contributing to a better understanding of the parasite-host relationships that occur in these environments. In addition, it also helps in the implementation of management, treatment, and control activities for the parasites that occur in these host species, contributing to the reduction in the number of zoonotic diseases in these parks (Fagiolini et al., 2010).
The present work aimed to report the occurrence of endoparasites in the feces of captive animals and in the animals that attended the Arruda Câmara Zoobotanical Park, João Pessoa, Paraíba, Brazil, as well as to identify the statistical differences between the percentages of the parasites found.

Ethical aspects
The project was submitted to the Ethics Committee on the Use of Animals in Research of the Universidade Federal Rural do Semi-Arido (UFERSA) (CEUA-UFERSA) and approved (Nº of opinion 11/2020). All the handling procedures of the animals followed the specific guidelines of the Brazilian College of Animal Experimentation.

Study area
The research was conducted at the Arruda Câmara Zoobotanical Park (ACZP), (CNPJ: 08.806.721/0001-03). This park is registered at the Brazilian Institute for the Environment and Renewable Natural Resources under the Registration No.
236567. It is popularly known as Bica, being located between the coordinates 292670mE and 293330mE, and between 9213107mN and 9214100mN, more precisely in the northern part of the municipality of João Pessoa. This park is set in a 26.4hectare Atlantic Forest fragment, being the home to native and exotic flora and fauna species, distributed among different areas available for visitation ( Figure 1).

Sampled animals
We analyzed 66 fecal samples from animals that were kept at the ACZP, João Pessoa-PB, as well as from wild animals that were attended, totaling 50 species. The fecal samples spontaneously eliminated by the animals were collected individually or in pools from the floor of the enclosure to avoid stressing the hosts and endangering the handlers of the animals.

Laboratory Analysis
The fecal samples were preserved in 5% formalin and sent to the Laboratory of Animal Parasitology (LAP) at the Universidade Federal Rural do Semi-Árido (UFERSA) for coproparasitological analysis. The techniques used were the direct method (Hoffmann, 1987), zinc sulfate flotation (Willis, 1921), spontaneous sedimentation (Hoffmann et al., 1934), and Mini-FLOTAC (Cringoli et al., 2012;2013;. The Mini-FLOTAC technique (Cringoli et al., 2012;2013;  Photographs of the endoparasites found were taken with a digital camera and the classification was made according to specific literature, such as Foreyt (2002).

Statistical analysis
The data were collected in a table and then transferred to the statistical program SPSS (Statistical Page for Social Sciences) version 23.0. They were expressed as simple frequency and percentage of parasites and compared among them for significant statistical differences using the binomial test for homogeneous proportions. The significance level was set at 5%.

Results
Of the total number of analyzed fecal samples ( Research, Society andDevelopment, v. 10, n. 13, e486101321255, 2021 (CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v10i13.21255 8 In the positive samples, the highest prevalence (p=0.001) was of Nematoda, with 80.5% (29/36). Among these worms, parasitic forms of the orders Strongylida (27.5%), Trichinellida (27.5%), and Rhabditida (37.9%) were found. Due to the similarity between the eggs of the order Strongylida, and since we used only morphological traits in this study, we decided in some cases to make an approximate identification of specimens from Ancylostomatidae and Strongylida. Trichinellida eggs of the genera Capillaria and Trichuris were observed. Among the Rhabditida, the following parasites were found: rhabditoid larvae; eggs of the superfamily Ascarididoidea, such as eggs of Toxascaris leonina, Toxocara sp, and unidentified eggs obtained from Crax fasciolata (suggested as Heterakis spp. or Ascaridia spp.); eggs of the superfamily Oxyuroidea, such as eggs of Syphacia sp. and Aspiculuris sp.; and eggs of the superfamily Strongyloidoidea, in Salvator merianae (suggested as Strongyloides sp. or Rhabdias sp.). In addition, eggs of Nematoda were found in Saimiri sciureus ( Table 2).
Cestodes were found in 19.4% of the animals analyzed (07/36), all of them belonging to the order Cyclophyllidea.
Thus, we recovered eggs from the Hymenolepididae family, such as Hymenolepis spp; from the Anoplocephalidae family, such as Bertiella spp; and eggs of unidentified Cestoda (suggested as Hymenolepis spp. and Paratriotaenia spp.) (Table 2, Figure   2). Balantidium, both being reported for the first time in Brazil. Generally, infections by endoparasites in free-living tapirids are asymptomatic, but signs of parasitic disease have been observed in captive animals. We also highlight the first record of Bertiella spp. in Alouatta caraya in the Northeast, Brazil. Clinical signs of this parasitic disease have not been demonstrated for this primate species, but the importance of this zoonosis has been reported, which may occur mainly in patients with direct or indirect contact with these animals, resulting in abdominal discomfort, diarrhea, gastroenteritis, and anorexia (Oliveira et al., 2011;Fernandes-Santos et al., 2020).
Further, this is the first occurrence of Hymenolepis spp. and Aspiculuris spp. as spurious parasites from reptiles in Brazil, exhibiting pseudoparasitism in these animals from the moment they ingest rodents, which are the definitive hosts.
Although these pseudoparasites do not infect them, reptiles can be regarded as dispersers of viable parasitic forms that may cause infection in their respective hosts, such as rodents, non-human primates, and humans. In addition, this study also highlights the parasitism by Entamoeba coli and Eimeria spp. in Iguana iguana. Although infection by the latter is usually asymptomatic, generally when Entamoeba coli affects the gastrointestinal system, it may cause necrotic enteritis, hepatitis and liver abscesses, lethargy, diarrhea, regurgitation, convulsions, depression, hematochezia, and death in Squamata (Rinaldi et al., 2012;Lima et al., 2017;2021).
Despite the great relevance of Zoological Parks and the struggle of these institutions to maintain the health of their animals, implementing prevention, diagnosis, and treatment programs, it is well known that many captive animals are housed close to each other, making parasitic infections inevitable. In addition, captive animals are often under considerable stress, which decreases their immunity and makes them more susceptible to infections. These parasites can pose a serious threat to the captive animals, occasionally causing fatalities. Many of these diseases, besides interfering with the welfare of the host species and affecting considerably their birth and mortality rates, may be zoonotic, posing a risk to animal handlers and animal care workers. In the present case, there was still the aggravating factor of the enclosures being within a forest reserve, which favors a greater contact of the animals with the droppings of other individuals, or even with intermediate hosts (Fagiolini et al., 2010;Oliveira et al., 2011;Snak et al., 2014;Schieber, M.C.;Štrkolcová, 2019;Dashe;Behanu, 2020;Patra et al., 2020).
The parasitological diagnosis of wild and exotic animals kept in captivity is essential to assist decisions related to their treatment, since they provide important information about the health of the herd and the immune resistance of the hosts. In addition, this information collaborates with the scientific community and contributes to provide ecological data for each of the species herein studied, favoring their conservation and preservation (Barros et al., 2017).
Although there are previous works that have carried out parasitological surveys in animals from zoos in Brazil, these studies, besides being very scarce, are much spaced. Typically, these investigations did not evaluate most of the animals kept at those zoobotanic parks. The present research was the pioneer, in Brazil, to perform the parasitological diagnosis of most of the vertebrate animals of the same zoobotanic park, with species from the Mammalia, Aves, Archelosauria, and Lepidosauria taxa.

Conclusions
A total of 54.5% of the tested animals were parasitized, which represents a risk to the health of both humans and animals, since many of these pathogens are also of medical importance. This research helps to broaden the ecological data and assists in the ex-situ conservation of wild and exotic animals. Based on the knowledge generated by this study, additional work is possible in order to understand the ecology of these endoparasites in the animals herein studied; investigate the potential consequences that these hosts may face in the wild; and the way parasitism might affect the welfare of host species in captivity and in the wild. Finally, this study contributes to the knowledge of endoparasites that may occur in zoos in Brazil and specifically in the Northeast region of this country, as well as for the species studied here.