Ticiana Zwarg

Hematological profile of Chelonia mydas (Testudines, Cheloniidae) according to the severity of fibropapillomatosis or its absence

Rossi S., Zwarg T., Sanches T.C., Cesar M.O., Werneck M.R. & Matushima E.R. 2009. Hematological profile of Chelonia mydas (Testudines, Cheloniidae) according to the severity of fibropapillomatosis or its absence. Pesquisa Veterina-

Flow cytometry as a tool in the evaluation of blood leukocyte function in Chelonia mydas (Linnaeus, 1758) (Testudines, Cheloniidae)

Chelonia mydas is a sea turtle that feeds and nests on the Brazilian coast and a disease called fibropapillomatosis is a threat to this species. Because of this, it is extremely necessary to determine a methodology that would enable the analysis of blood leukocyte function in these sea turtles. In order to achieve this aim, blood samples were collected from C. mydas with or without fibropapillomas captured on the São Paulo north coast. Blood samples were placed in tubes containing sodium heparin and were transported under refrigeration to the laboratory in sterile RPMI 1640 cell culture medium. Leukocytes were separated by density gradient using Ficoll-PaqueTM Plus, Amershan Biociences. The following stimuli were applied in the assessment of leukocyte function: Phorbol Miristate-Acetate (PMA) for oxidative burst activity evaluation and Zymosan A (Saccharomyces cerevisiae) Bio Particles, Alexa Fluor 594 conjugate for phagocytosis evaluation. Three cell populations were identified: heterophils, monocytes and lymphocytes. Monocytes were the cells responsible for phagocytosis and oxidative burst.

Hematological and histopathological evaluation of wildlife green turtles (Chelonia mydas) with and without fibropapilloma from the north coast of São Paulo State, Brazil

Blood profiles were determined in 47 juvenile green turtles, Chelonia mydas, from Sao Paulo northern coast, Brazil. Twenty-nine were affected by fibropapillomas and 18 were tumor free. Complete gross and histopathologic examinations of the fibropapillo were performed in 21 green turtles. Biometrical data, size, location and amount of tumors were recorded. The papillomas varied in morphology, location, size, color and texture. We found hyperplastic stroma, rich in blood vessels and connective tissue with increase in thickness of the dermis. The tumors w0ere classified as papillomas or fibropapillomas according to their epithelial and/or stromal proliferation. The lowest Mean Corpuscular Hemoglobin (HCM) values were observed in affected turtles.

Hematologic reference values for clinically healthy captive golden conures (Guaruba guarouba).

SUMMARY. Golden conures or ararajubas (Guaruba guarouba) are endangered parrots endemic to the Brazilian Amazon forest. Body mass, blood cell counts, and total plasma protein were determined for 70 clinically healthy golden conures captive at zoologic parks and private breeder facilities in Brazil. Hematologic results (mean ± SD) were: Erythrocytes 3.6 ± 0.5 × 106 cells/mm3, hemoglobin 12.8 ± 1.4 g/dl, packed cell volume 46 ± 3.8%, mean corpuscular volume 132 ± 20 fl, mean corpuscular hemoglobin (MCH) 36 ± 5.7 pg, mean corpuscular hemoglobin concentration (MCHC) 28 ± 3.5%, thrombocytes 26.3 ± 9.3 × 103 cells/mm3, leukocytes 11.9 ± 4.5 × 103 cells/mm3, heterophils 6284 ± 2715 cells/mm3, lymphocytes 5473 ± 2408 cells/mm3, monocytes 113 ± 162 cells/mm3, eosinophils 10 ± 42 cells/mm3, basophils 27 ± 64 cells/mm3. Body mass was 254 ± 24.9 g and total plasma protein (TPP) was 3.54 ± 0.58 g/dl. No statistical differences were observed between genders within age groups. Differences between juveniles (J) and adults (A) were identified for TPP (J < A), MCH (J > A), and MCHC (J > A). These results provide reliable reference values for the clinical interpretation of hematologic results for the species. Hematology may be an important tool for population health investigations on free-ranging golden conure populations and will also be essential to survey the health of release candidates in future reintroduction programs.

Diversity of Sarcocystis spp shed by opossums in Brazil inferred with phylogenetic analysis of DNA coding ITS1, cytochrome B, and surface antigens.

Although few species of Sarcocystis are known to use marsupials of the genus Didelphis as definitive host, an extensive diversity of alleles of surface antigen genes (sag2, sag3, and sag4) has been described in samples of didelphid opossums in Brazil. In this work, we studied 25 samples of Sarcocystis derived from gastrointestinal tract of opossums of the genus Didelphis by accessing the variability of sag2, sag3, sag4, gene encoding cytochrome b (cytB) and first internal transcribed spacer (ITS1). Reference samples of Sarcocystis neurona (SN138) and Sarcocystis falcatula (SF1) maintained in cell culture were also analyzed. We found four allele variants of cytB, seven allele variants of ITS1, 10 allele variants of sag2, 13 allele variants of sag3, and 6 allele variants of sag4. None of the sporocyst-derived sequences obtained from Brazilian opossums revealed 100% identity to SN138 at cytB gene, nor to SN138 or SF1 at ITS1 locus. In addition, none of the sag alleles were found identical to either SF1 or SN138 homologous sequences, and a high number of new sag allele types were found other than those previously described in Brazil. Out of ten sag2 alleles, four are novel, while eight out of 13 sag3 alleles are novel and one out of six sag4 alleles is novel. Further studies are needed to clarify if such a vast repertoire of allele variants of Sarcocystis is the consequence of re-assortments driven by sexual exchange, in order to form individuals with highly diverse characteristics, such as pathogenicity, host spectrum, among others or if it only represents allele variants of different species with different biological traits.

Detection of bacteria and fungi and assessment of the molecular aspects and resistance of Escherichia coli isolated from confiscated passerines intended for reintroduction programs

Many native bird species are currently considered rare in Brazil because they have been indiscriminately collected by animal traffickers and commercialized, leading to dwindling numbers in their natural habitats. Confiscated animals are at times destined for reintroduction programs that must ensure these animals do not pose a risk to native populations. Healthy or sick wild passerines may carry a great diversity of microorganisms. Therefore, knowledge of the sanitary status of confiscated animals destined for reintroduction is critical to assess whether these animals act as microorganism carriers and to investigate the epidemiology of transmissible diseases, a crucial aspect for animal and human health preservation. This study examined the occurrence of aerobic and facultative anaerobic bacteria and fungi in cloacal swabs collected from wild confiscated passerines intended for reintroduction programs. In vitro susceptibility tests of the most frequent isolates as well as studies of the molecular aspects of Escherichia coli isolates were also performed. There was microorganism growth in 62.5% of 253 samples. The microorganisms that were most frequently isolated were Staphylococcus spp. (15.0%), Micrococcus spp. (11.5%), E. coli (10.7%) and Klebsiella spp. (10.7%). Fifteen bacteria genera and seven fungi genera were isolated. Multidrug-resistance to antimicrobials was observed in Staphylococcus spp., Micrococcus spp., E. coli and Klebsiella spp. isolates. The high occurrence of Enterobacteria observed is possibly related to the sanitary conditions in which confiscated animals are usually kept. One E. coli sample (out of 27 isolates) was positive for the S-fimbrial adhesion encoding gene (sfa). Considering the low occurrence of genes that encode virulence factors, confiscated passerines may represent a low risk for the potential transmission of EPEC, APEC, UPEC and NMEC isolates to other animals or humans. The potential risk of intra- or inter-specific transmission of multidrug-resistant isolates and the introduction of these microorganisms into the environment must be considered, although there are still therapeutic alternatives for treatment of these animals among the antimicrobials which were tested. The stress and poor hygiene conditions imposed on animals during trafficking may have caused their contamination by multidrug-resistant agents transmitted by humans or by the precarious environment to which they were subjected. Risks related to the dissemination of Salmonella spp., Cryptococcus spp. and Candida spp. are low when reintroduction programs are considered.

Hematological parameters and the variations resulting from stress of Alouatta caraya during a wildlife rescue program in Brazil

Deforestation and habitat fragmentation are major threats to the conservation status of New World primates, such as the howler monkey (Alouatta caraya) in Brazil, where vegetation destruction is often associated with projects such as Small Hydro Power Plant (SHP) construction. The resulting stress from the implementation of this type of enterprise may be a factor that influences individual susceptibility against pathogens and can determine the success or failure of mitigation measures proposed by responsible and/or requested companies by environmental agencies. To perform health monitoring and to understand physiological changes that ensued from the stress of capturing and keeping primates in captivity, we determined the blood profile of A. caraya rescued at the time of capture (N = 15) and in captivity (N = 11). Complete blood counts (CBCs) performed at the time of capture showed hematological changes compatible with acute stress, such as evident neutrophilic leukocytosis (WBCs; females = 20.48 ± 7.26; males = 18.78 ± 10.39 × 109/L). In captivity, monocytosis was observed in both sexes (females = 1.34 ± 0.26; males = 0.39 ± 0.32 × 109/L; U‐test, P = 0.01), suggesting chronic stress. In females, there was also statistically significant eosinopenia (0.28 ± 0.10 × 109/L; U‐test, P = 0.01) and hematocrit increases (39.00 ± 1.41%; U‐test, P = 0.01). Thus, data shows the impact of both capture related acute stress as well as captivity chronic stress, suggesting that primate management in the implementation of these projects causes significant changes in physiological parameters and, consequently, animal health. Whereas chronic stress is an inducer of immunosuppression and susceptibility to pathogen factors, monitoring hematological parameters in captive animals can act as an indicator of health status, contributing to the success of management and conservation wildlife programs. Am. J. Primatol. 77:246–253, 2015.

Normal Plasma Cholinesterase Activity of Neotropical Falconiformes and Strigiformes

Abstract Raptors are highly susceptible to poisoning by anticholinesterase drugs and pesticides, such as organophosphates and carbamates, due to their carnivorous and/or insectivorous diets. Measurement of plasma cholinesterase inhibition is a proxy indicator of exposure to these agents; however, prior knowledge of the normal plasma cholinesterase activity of healthy birds is required for interpretation of results. Such normal values have not yet been established for most species of neotropical raptors. We collected blood samples from 133 raptors, including eleven species of Falconiformes (n  =  45) and seven species of Strigiformes (n  =  88). The birds had no clinical signs or history of possible exposure to anticholinesterase drugs and were housed in rehabilitation centers (free-ranging animals) and zoos (captive animals) in the state of São Paulo, southeastern Brazil. We found that plasma cholinesterase activity, as measured by the Ellman method, had a logarithmic distribution with a median of 0.58 µmols/mL for Falconiformes (quartiles: Q1  =  0.43; Q3  =  0.73) and 0.96 µmols/mL for Strigiformes (Q1  =  0.78; Q3  =  1.25). There were no significant differences between sex or age classes within Falconiformes or Strigiformes. Captive Strigiformes in zoos had lower plasma cholinesterase activity values than birds housed in the rehabilitation centers, a pattern that did not occur in Falconiformes. These results provide reference values of plasma cholinesterase activity for the future interpretation of suspected pesticide poisoning in neotropical raptors.

Parotid Salivary Gland Basal Cell Adenocarcinoma in a Big-eared Opossum (Didelphis aurita)

The opossum (family Didelphidae) is a marsupial endemic to the Americas. Apart from the South American short-tailed opossum (Monodelphis domestica) and the Virginia opossum (Didelphis virginiana), there is considerable lack of knowledge about the health and diseases of most opossum species. Among these, the big-eared opossum (Didelphis aurita) is found in Argentina, Brazil and Paraguay. Natural and experimental studies have shown this species to be susceptible to infectious agents with zoonotic potential and the animals may play a role in transmission of such agents. However, neoplasia appears to be uncommon in this species. We describe the gross, microscopical and immunohistochemical features of a parotid salivary gland basal cell adenocarcinoma in a free-living big-eared opossum. This case represents the first report of salivary gland neoplasia in opossums.