Concentrations of polycyclic aromatic hydrocarbons in liver samples of juvenile green sea turtles from Brazil: Can these compounds play a role in the development of fibropapillomatosis?

Fibropapillomatosis (FP) poses a significant threat to the conservation of green sea turtles (Chelonia mydas). Polycyclic aromatic hydrocarbons-PAHs are considered mutagenic, carcinogenic and toxic, and can act as cofactor of this disease. In order to evaluate possible differences between green sea turtles with and without FP, we monitored 15 PAHs in liver samples of 44 specimens (24 with FP) captured in Brazil. We detected eight PAHs and quantified phenanthrene in all green sea turtles with FP. Specimens without FP presented lower values than the tumored ones (1.48 ng g-1 and 17.35 ng g-1, respectively; p < 0.0001). There were no significant differences between tumored and non-tumored specimens, among studied areas, or Southwest Atlantic Fibropapillomatosis Score. Even though we found higher concentrations in the liver samples of green sea turtles with FP, further studies are necessary to confirm if these pollutants are involved in the pathogenesis of the disease.

Challenges in Evaluating the Severity of Fibropapillomatosis: A Proposal for Objective Index and Score System for Green Sea Turtles (Chelonia mydas) in Brazil.

Fibropapillomatosis (FP) is a neoplastic disease that affects marine turtles worldwide, especially green sea turtles (Chelonia mydas). FP tumors can develop on the body surface of marine turtles and also internally in the oral cavity and viscera. Depending on their quantity, size and anatomical distribution, these tumors can interfere with hydrodynamics and the ability to feed, hence scoring systems have been proposed in an attempt to quantify the clinical manifestation of FP. In order to establish a new scoring system adapted to geographic regions, we examined 214 juvenile green sea turtles with FP caught or rescued at Brazilian feeding areas, counted their 7466 tumors and classified them in relation to their size and anatomical distribution. The patterns in quantity, size and distribution of tumors revealed interesting aspects in the clinical manifestation of FP in specimens studied in Brazil, and that FP scoring systems developed for other areas might not perform adequately when applied to sea turtles on the Southwest Atlantic Ocean. We therefore propose a novel method to evaluate the clinical manifestation of FP: fibropapillomatosis index (FPI) that provides the Southwest Atlantic fibropapillomatosis score (FPSSWA). In combination, these indexing and scoring systems allow for a more objective, rapid and detailed evaluation of the severity of FP in green sea turtles. While primarily designed for the clinical manifestation of FP currently witnessed in our dataset, this index and the score system can be adapted for other areas and compare the characteristics of the disease across regions. In conclusion, scoring systems to classify the severity of FP can assist our understanding on the environmental factors that modulate its development and its impacts on the individual and population health of green sea turtles.

Health evaluation and survey of zoonotic pathogens in free-ranging capybaras (Hydrochoerus hydrochaeris).

Capybaras (Hydrochoerus hydrochaeris) are the world's largest rodents and play an epidemiologic role in the transmission of zoonotic pathogens, including the causative agents of Brazilian spotted fever, leptospirosis, and others. We surveyed the health of 31 free-ranging capybaras at the Alberto Löfgren State Park, São Paulo, Brazil using a variety of diagnostic methods. Hematology and serum chemistry were consistent with mild malnutrition and parasitism but did not indicate severe physiologic imbalance or disease. All animals were serologically negative for Rickettsia rickettsii, Leishmania spp., and Trypanosoma sp., but antibodies against rabies virus (71%), Leptospira sp. (26%), and Toxoplasma sp. (23%) were detected. Salmonella sp. was not cultured from fecal samples. Frequently cultured enterobacteria included Escherichia coli (61%), Enterococcus casseiflavus (35%), Enterococcus faecalis (35%), Enterobacter aerogenes (32%), Klebisella pneumoniae (32%), and Serratia marcescens (32%). No potentially pathogenic fungi were cultured from hair samples. Fecal parasitology revealed infection by Protozoophaga sp. (58%), Viannella spp. (23%), Strongyloides spp. (10%), and Ancilostomatidae (10%). A total of 218 ticks was retrieved from the animals: Amblyomma sp. larvae and nymphs (43%), A. dubitatum adults (52%), and A. cajennense adults (5%). The capybaras were free from most potentially zoonotic pathogens evaluated; however, the presence of Amblyomma spp. ticks (potential vectors of Rickettsia spp.) and indirect evidence of exposure to the rabies virus, Leptospira sp., and Toxoplasma sp. warrant the maintenance of public health programs and wildlife health monitoring.

Enterobacterial colonization in captive red-eared sliders (Trachemys scripta elegans).

The handling of turtles and other reptiles can be associated with risk of pathogenic enterobacteria transmission, mainly Salmonella spp. The aim of this study was to identify the enterobacteria in cloacal swabs of 39 red-eared sliders (Trachemys scripta elegans). Cloacal swabs from 39 captive individuals were analyzed. After sample enrichment in brain-heart infusion broth and 1% peptone water, bacterial isolation was performed through cultivation in blood, MacConkey and xylose lysine desoxycholate agar. Bacterial identification was achieved through conventional tests and automated turbidity analysis. The results indicated the growth of Kluyvera ascorbata (38/39), Leclercia adecarboxylata (37/39), Raoultella planticola (30/39), Citrobacter freundii (20/39), Proteus spp. (15/39), and Escherichia coli (5/39). Salmonella spp. were not detected. The intestinal enterobacteria identified inthis study differed from that reported in the literature for other reptiles.

Genetic similarity between APEC and Escherichia coli strains isolated from Guaruba guarouba in a survey on healthy captive psittacine birds / Similaridade genética entre APEC e cepas de Escherichia coli isoladas de Guaruba guarouba em um estudo com psitacídeos hígidos de cativeiro

The role of psittacine birds as a reservoir of avian pathogenic Escherichia coli (APEC) is not known but would be helpful in understanding the human – animal interface, since the enteric microbiota of these birds consists of Gram positive bacteria. The aim of this study was to identify the presence of APEC in feces of clinically healthy Guaruba guarouba. To do this, we isolated and analyzed E. coli from cloacal fecal samples taken from 87 psittacine birds from six zoologic parks, three commercial breeders and one conservation breeder. Of the 87 birds examined, 46 (52.87%) presented E. coli in feces. The presence of the following eight virulence genes was determined by the polymerase chain reaction (PCR): irp2, iucD, iss, vat, cvi/cva, tsh, astA, and papC, and 29 (63.04%) of 46 E. coli isolates tested were positive at least one of the eight genes studied. The frequency of virulence genes observed in isolates of E. coli were 32.6% (15/46) irp2, 26% (12/46) iucD, 19.5% iss (9/46), 17.4% vat (8/46), 17.4% cvi/cva (8/46), 8.7% tsh (4/46), 4.4% astA (2/46) and 0% papC (0/46). The isolates were grouped in 13 genotypic profiles according to virulence gene combinations, but only 2 isolates were classified as APEC, with the pattern iuc, iss, cvi/cva, irp + and iuc, iss, cvi/cva, irp, tsh, vat +. This study reveals the presence of APEC in clinically healthy captive G. guarouba, suggesting that these psittacine birds may act as reservoir for pathogenic microorganisms. Epidemiological studies are needed to determine the relevance of this species as a reservoir and the implications for conservation of endangered species G. guarouba.

O papel dos psitacídeos como reservatório de Escherichia coli patogênicas para aves (APEC) não é conhecido, mas será útil para a compreensão da interface humano-animal, uma vez que a microbiota entérica destas aves é composta por bactérias Gram-positivas. O objetivo deste estudo foi identificar a presença de APEC em fezes de Guaruba guarouba clinicamente saudáveis. Para isso, foram isoladas e analisadas E. coli presentes em fezes cloacais coletadas de 87 psitacídeos, alojados em seis zoológicos, três criatórios comerciais e um criatório conservacionista. Das 87 aves examinadas, 46 (52,87%) apresentaram E. coli nas fezes. A presença de oito genes de virulência foi determinada pela reação em cadeia pela polimerase (PCR): irp2, iucD, iss, vat, cvi/cva, tsh, astA, e papC, e 29 (63,04%) dos 46 isolados foram positivos para pelo menos um dos oito genes estudados. A frequência dos genes de virulência observada nos isolados de E. coli foi 32.6% (15/46) irp2, 26% (12/46) iucD, 19.5% iss (9/46), 17.4% vat (8/46), 17.4% cvi/cva (8/46), 8.7% tsh (4/46), 4.4% astA (2/46) e 0% papC (0/46). Os isolados foram agrupados em 13 perfis genotípicos de acordo com combinações de genes de virulência, mas apenas duas amostras foram classificadas como APEC, com o perfil iuc, iss, cvi/cva, irp + e iuc, iss, cvi/cva, irp, tsh, iuc, iss, +. Este estudo revela a presença de APEC em aves de cativeiro (G. guarouba) clinicamente saudáveis, sugerindo que estes psitacídeos possam atuar como reservatórios de micro-organismos patogênicos. Estudos epidemiológicos são necessários para determinar a relevância desta espécie como reservatório e as implicações para a conservação da espécie ameaçada G. guarouba.

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

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 x 10(6) 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 x 10(3) cells/mm3, leukocytes 11.9 +/- 4.5 x 10(3) 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.