Molecular detection of hemogregarines and haemosporidians in Brazilian free-living testudines.

Morphological and molecular techniques were used to investigate the presence of hemogregarines and haemosporidians in biological samples of free-living Geoffroy's side-necked turtles (Phrynops geoffroanus) and Giant Amazon turtles (Podocnemis expansa) from Brazil. No evolutionary form of haemosporidians or hemogregarines were observed in the blood smears of 83 P. geoffroanus samples, and there were no meronts in the histological sections of 31 necropsied P. geoffroanus samples. All DNA samples extracted from P. geoffroanus tissues and blood aliquots were negative in haemosporidian PCR assays (based on the mitochondrial cytochrome b gene) and hemogregarine PCR assays (based on the 18S rRNA gene). In the analysis of blood smears of all seven Podocnemis expansa evaluated, gametocytes of hemogregarines were observed. The seven P. expansa were negative in the haemosporidian PCR assays. Moreover, hemogregarine DNA was detected in blood samples from all of the sampled P. expansa. The phylogenetic maximum likelihood inference and probabilistic Bayesian inference revealed five closely related genotypes that formed a monophyletic group. There was also a sister group to the lineage that consisted of Haemogregarina spp. of freshwater turtles from Canada, Italy, Mozambique, Kenya, Gabon, Vietnam, and China. The findings suggest that free-living P. expansa were parasitized by a new genotype or even a possible new species of the genus Haemogregarina. Haemosporidians and hemogregarines are not frequently found in P. geoffroanus in the studied region under the local conditions of that period.

Arthropod-borne agents in wild Orinoco geese (Neochen jubata) in Brazil.

Although Orinoco goose (Neochen jubata) is an anatid species widely distributed in South America, scarce are the reports on the occurrence of arthropod-borne pathogens in this avian species. The present work aimed to verify, by serological and molecular methods, the occurrence of haemosporida piroplasmids and Anaplasmataceae agents in wild Orinoco geese captured in Brazil. Between 2010 and 2014, 62 blood samples were collected from free-living geese captured in the Araguaia River, Goiás State, Brazil. Six geese (10%) were seropositive for Anaplasma phagocytophilum, showing titers ranging from 40 and 80. Twenty out of 62 blood samples (32.25%) were positive in nested PCR for hemosporidia (cytochrome b gene). Fifteen and five sequences shared identity with Haemoproteus and Plasmodium, respectively. Six out of 62 blood samples (9.68%) were positive in nested PCR for Babesia spp. (18S rRNA gene); one sequence showed to be closely related to Babesia vogeli. Thirty (48.38%) out of 62 Orinoco geese blood samples were positive in nested cPCR assays for Anaplasmataceae agents (16S rRNA gene): three for Anaplasma spp. and 27 for Ehrlichia. Six geese were simultaneously positive to Haemoproteus and Ehrlichia; three animals were co-positive to different Ehrlichia species/genotypes; and one goose sample was positive for both Anaplasma and Ehrlichia. The present work showed the occurrence of Ehrlichia, Anaplasma, Babesia, Plasmodium, and Haemoproteus species in free-living N. jubata in Brazil. The threat of these arthropod-borne pathogens in Orinoco goose's fitness, especially during the breading season, should be assessed in the future.

Isolation of Leptospira borgpetersenii in synanthropic Didelphis albiventris in Jaboticabal, São Paulo, Brazil / Isolamento de Leptospira borgpetersenii em Didelphis albiventris sinantrópicos em Jaboticabal, São Paulo, Brasil

Leptospirosis is a waterborne disease and, therefore, stands out for the possibility of environmental contamination, the cross transmission between domestic and wild animals and humans. Opossum species are important reservoirs of this disease making them potential pathogen spreaders. Aiming to verify the presence of Leptospira spp. and the antibodies against Leptospira spp. in the Campus of São Paulo State University, in Jaboticabal, São Paulo, Brazil, freeliving wild life opossum (Didelphis albiventris) were captured for blood and urine sampling. Serological analysis was performed Microscopic Agglutination Test (MAT). Aliquots of urine were seeded in media Ellinghausen-McCullough- Johnson-Harris (EMJH) and Fletcher without antibiotics. The samples in which there was growth of leptospires were forwarded to the Leptospirosis Laboratory of the Institute of Pathobiology in the National Institute of Agricultural Technology, Buenos Aires, Argentina and were genotyped using Multiple Locus Variable number tandem repeat Analysis (MLVA). Of the 15 analyzed animals, nine (60.0%) were reactant to Patoc serovar. The pathogenic specie Leptospira borgpetersenii was isolated and identified in three Didelphis albiventris. The isolation findings of pathogenic specie Leptopsira borgpetersenii in the urine culture of three Didelphis albiventris in a university campus are a major discovery in the area of preventive veterinary medicine and public health and open a discussion about the important role of free-living wild animals as reservoirs of this agent to domestic animals and humans, a condition that serves as a warning for the improvement of health practices...

A leptospirose é uma zoonose de veiculação hídrica e, portanto, se destaca pela possibilidade de contaminação ambiental, o que facilita a transmissão cruzada entre animais domésticos, selvagens e humanos. Espécies de gambás são importantes reservatórios dessa enfermidade, tornando-os potenciais disseminadores do agente. Com o objetivo de verificar a presença de Leptospira spp. e de anticorpos contra Leptospira spp. no Campus da Universidade Estadual Paulista, em Jaboticabal, foram capturados gambás (Didelphis albiventris) de vida livre para a colheita de amostras de sangue e de urina. As análises sorológicas foram efetuadas pela técnica de Soroaglutinação Microscópica (SAM). Alíquotas de urina foram semeadas nos meios Ellinghausen-McCullough-Johnson-Harris (EMJH) e Fletcher sem antibióticos. As amostras que apresentaram crescimento de espiroquetas foram levadas ao Laboratório de Leptospirose do Instituto de Patobiologia, no Instituto Nacional de Tecnologia Agropecuaria, Buenos Aires, Argentina e foram genotipadas com a técnica de Múltiplos Locus de Números Variáveis de Repetição em Tandem (MLVA). Dos 15 animais examinados pela SAM, nove (60,0%) foram reagentes à sorovariedade Patoc. Foi isolada e identificada a espécie patogênica Leptospira bosrpetersenii de três Didelphis albiventris. Os achados de isolamento da espécie patogênica Leptospira borgpetersenii na cultura de urina de três Didelphis albiventris são um grande descobrimento para as áreas da medicina veterinária preventiva e da saúde pública e reforçam a discussão sobre o importante papel dos animais selvagens de vida livre como reservatórios desse agente para animais domésticos e seres humanos, situação que serve de alerta para melhorias nas práticas sanitárias...

Association of acepromazine with propofol in giant Amazon turtles Podocnemis expansa reared in captivity.

PURPOSE: To evaluate the effects of different concentrations of an anesthetic association in giant amazon turtles (Podocnemis expansa). METHODS: Twenty healthy P. expansa of both sexes weighing between 1.0 and 1.5kg commercially bred in the Araguaia River Valley, Goias, Brazil, were separated into two groups (G1 n=10 and G2 n=10). Each group received a respective protocol: P1= acepromazine (0.5 mg/kg IM) and propofol (5 mg/kg IV) and P2 = acepromazine (0.5 mg/kg IM) and propofol (10 mg/kg IV). The acepromazine was administered in the left thoracic member and the propofol in the cervical vertebral sinus. Assessments were made of the anesthetic parameters of locomotion, muscle relaxation, response to pain stimuli in the right thoracic and pelvic members and heartbeat. RESULTS: The anesthetic induction time was the same for both protocols (P1 and P2); however the P2 effects were of a longer duration. CONCLUSION: The sedation achieved with both protocols (P1 and P2) were satisfactory for the biological sample collection, physical examinations and minor surgeries on this species.

Association of acepromazine with propofol in giant amazon turtles Podocnemis expansa reared in captivity / Associação da acepromazina com propofol em tartarugas-da-amazônia Podocnemis expansa criadas em cativeiro

PURPOSE: To evaluate the effects of different concentrations of an anesthetic association in giant amazon turtles (Podocnemis expansa). METHODS: Twenty healthy P. expansa of both sexes weighing between 1.0 and 1.5kg commercially bred in the Araguaia River Valley, Goias, Brazil, were separated into two groups (G1 n=10 and G2 n=10). Each group received a respective protocol: P1= acepromazine (0.5 mg/kg IM) and propofol (5 mg/kg IV) and P2 = acepromazine (0.5 mg/kg IM) and propofol (10 mg/kg IV). The acepromazine was administered in the left thoracic member and the propofol in the cervical vertebral sinus. Assessments were made of the anesthetic parameters of locomotion, muscle relaxation, response to pain stimuli in the right thoracic and pelvic members and heartbeat. RESULTS: The anesthetic induction time was the same for both protocols (P1 and P2); however the P2 effects were of a longer duration. CONCLUSION: The sedation achieved with both protocols (P1 and P2) were satisfactory for the biological sample collection, physical examinations and minor surgeries on this species.

OBJETIVO: Avaliar os efeitos de uma associação anestésica com diferentes concentrações em tartarugas-da-amazônia (Podocnemis expansa). MÉTODOS: Vinte P. expansa, hígidas, de ambos os sexos, com massa corporal entre 1,0 e 1,5 kg, de um criatório comercial localizado no vale do rio Araguaia, Goiás, Brasil, foram distribuídas em dois grupos (G1 n=10 e G2 n=10). Cada grupo recebeu um protocolo sendo: P1 = acepromazina (0,5 mg/kg IM) e propofol (5 mg/kg IV) e P2 = acepromazina (0,5 mg/kg IM) e propofol (10 mg/kg IV), aplicados nos grupos G1 e G2, respectivamente. A acepromazina foi aplicada no membro torácico esquerdo e o propofol no seio vertebral cervical. Foram avaliados os parâmetros anestésicos: locomoção, relaxamento muscular, resposta aos estímulos dolorosos no membro torácico direito e nos membros pelvinos e frequência cardíaca. RESULTADOS: O tempo de indução anestésica foi o mesmo para ambos os protocolos (P1 e P2), porém o P2 apresentou efeitos mais duradouros. CONCLUSÃO: As sedações obtidas por esses protocolos (P1 e P2) foram satisfatórias para a colheita de amostras biológicas, exames físicos e realização de pequenos procedimentos cirúrgicos nesta espécie.

Migratory and carnivorous birds in Brazil: reservoirs for Anaplasma and Ehrlichia species?

In order to investigate new hosts for Anaplasmataceae agents in Brazil, we collected blood samples from 21 wild birds. Using molecular techniques, we detected the presence of Anaplasma phagocytophilum, Ehrlichia chaffeensis, and an Ehrlichia species closely related to Ehrlichia canis in carnivorous avian blood samples. In addition, an Ehrlichia species closely related to an Ehrlichia species found in wild felines in Brazil was also detected in a goose blood sample. Wild birds may play a role as carriers of Anaplasmataceae agents in Brazil.

Association of midazolam with ketamine in giant Amazon river turtles Podocnemis expansa breed in captivity.

PURPOSE: Evaluate the effects of two anesthetic associations in giant Amazon river turtles (P. expansa). METHODS: Twenty P. expansa, healthy, of both sexes, with weights between 1.0 and 1.5 kg of a commercial breeding facility located in the valley of the Araguaia River, Goiás, Brazil, were divided into two groups ( G1 n = 10 and G2 n = 10). Each group received a protocol being: P1 = midazolam (2 mg/kg IM) and ketamine (20 mg/kg IM) and P2 = midazolam (2 mg/kg IM) and ketamine (60 mg/kg IM), applied on G1 and G2, respectively. The drugs were applied in the left forelimb. The clinical parameters evaluated were: locomotion, muscle relaxation, response to pain stimuli in the right thoracic and pelvic members and heart rate. These assessments were made at time 0 (immediately after injection) and times of 5, 10, 20, 30, 45, 60, 90, 120, 150 and 180 minutes after the injections. RESULTS: Group 2 showed a higher heart rate than G1 and more rapid and prolonged immobilization. CONCLUSION: The sedation scores obtained by these protocols (P1 and P2) were satisfactory, with possible pharmacological contention for collecting biological samples and physical examination in P. expansa.

Association of midazolam with ketamine in giant Amazon river turtles Podocnemis expansa breed in captivity / Associação de midazolam com cetamina em tartarugas da Amazônia Podocnemis expansa de cativeiro

PURPOSE: Evaluate the effects of two anesthetic associations in giant Amazon river turtles (P. expansa). METHODS: Twenty P. expansa, healthy, of both sexes, with weights between 1.0 and 1.5 kg of a commercial breeding facility located in the valley of the Araguaia River, Goiás, Brazil, were divided into two groups ( G1 n = 10 and G2 n = 10). Each group received a protocol being: P1 = midazolam (2 mg/kg IM) and ketamine (20 mg/kg IM) and P2 = midazolam (2 mg/kg IM) and ketamine (60 mg/kg IM), applied on G1 and G2, respectively. The drugs were applied in the left forelimb. The clinical parameters evaluated were: locomotion, muscle relaxation, response to pain stimuli in the right thoracic and pelvic members and heart rate. These assessments were made at time 0 (immediately after injection) and times of 5, 10, 20, 30, 45, 60, 90, 120, 150 and 180 minutes after the injections. RESULTS: Group 2 showed a higher heart rate than G1 and more rapid and prolonged immobilization. CONCLUSION: The sedation scores obtained by these protocols (P1 and P2) were satisfactory, with possible pharmacological contention for collecting biological samples and physical examination in P. expansa.

OBJETIVO: Avaliar os efeitos de duas associações anestésicas em tartarugas da Amazônia em (Podocnemis expansa). MÉTODOS: Vinte P. expansa, hígidas, de ambos os sexos, com massa corporal entre 1,0 e 1,5 kg, de um criatório comercial localizado no vale do rio Araguaia, Goiás, Brasil, foram distribuídas em dois grupos (G1 n=10 e G2 n=10). Cada grupo recebeu um protocolo sendo: P1 = midazolam (2 mg/kg IM) com cetamina (20 mg/kg IM) e P2 = midazolam (2 mg/kg IM) com cetamina (60 mg/kg IM), aplicados nos grupos G1 e G2, respectivamente. Os fármacos foram aplicados no membro torácico esquerdo. Os parâmetros clínicos avaliados foram: locomoção, relaxamento muscular, resposta aos estímulos dolorosos nos membros torácico direito e pelvinos e freqüência cardíaca. Essas avaliações foram feitas no tempo 0 (imediatamente após a injeção) e nos tempos 5, 10, 20, 30, 45, 60, 90, 120, 150 e 180 minutos após as injeções. RESULTADOS: O G2 apresentou maior freqüência cardíaca que o G1 e imobilização mais rápida e prolongada. CONCLUSÃO: As sedações obtidas por esses protocolos (P1 e P2) foram satisfatórias, sendo possível a contenção farmacológica para a coleta de amostras biológicas e exame físico em P. expansa.

The use of rocuronium in giant Amazon turtle Podocnemis expansa (Schweigger, 1812) (Testudines, Podocnemididae).

PURPOSE: To determine whether rocuronium would provide safe, short-term immobilization in Podocnemis expansa. METHODS: Twenty P. expansa, weighing on average 1.59 +/- 0.28 kg, were subjected to two protocols: G1 0.25 mg/kg IM of rocuronium and 0.07 mg/kg IM of neostigmine, while G2 received 0.50 mg/kg IM of rocuronium and 0.07 mg/kg IM of neostigmine. The drugs were applied, respectively, in the left and right thoracic members. Assessments were made of the anesthetic parameters of respiratory frequency, heartbeat, righting reflex, cloacal relaxation, palpebral and pupilar reflexes, easy handling, muscle relaxation, locomotion, response to pain stimuli in the right thoracic members, pelvic members and tail, ambient humidity and temperature. RESULTS: They were not found statistical differences between the dosages for the majority of the assessments. G1 was as efficient as G2. A consistent neuromuscular blockade effect was recorded 12 +/- 4.21 minutes in G1 and G2. All the animals were recovered in 150 minutes. CONCLUSIONS: Administration of rocuronium at dose of 0.25 to 0.5 mg/kg IM is a safe and effective adjunct to clinical proceedings or pre-anesthetics in P. expansa. Because rocuronium does not provide any analgesic or sedative effects, the duration of neuromuscular blockade without anesthesia should be minimized to avoid undue stress.

The use of rocuronium in giant Amazon turtle Podocnemis expansa (Schweigger, 1812) (Testudines, Podocnemididae) / Uso do rocurônio em tartaruga da Amazônia Podocnemis expansa (Schweigger, 1812) (Testudines, Podocnemididae)

PURPOSE: To determine whether rocuronium would provide safe, short-term immobilization in Podocnemis expansa. METHODS: Twenty P. expansa, weighing on average 1.59 ± 0.28 kg, were subjected to two protocols: G1 0.25 mg/kg IM of rocuronium and 0.07 mg/kg IM of neostigmine, while G2 received 0.50 mg/kg IM of rocuronium and 0.07 mg/kg IM of neostigmine. The drugs were applied, respectively, in the left and right thoracic members. Assessments were made of the anesthetic parameters of respiratory frequency, heartbeat, righting reflex, cloacal relaxation, palpebral and pupilar reflexes, easy handling, muscle relaxation, locomotion, response to pain stimuli in the right thoracic members, pelvic members and tail, ambient humidity and temperature. RESULTS: They were not found statistical differences between the dosages for the majority of the assessments. G1 was as efficient as G2. A consistent neuromuscular blockade effect was recorded 12 ± 4.21 minutes in G1 and G2. All the animals were recovered in 150 minutes. CONCLUSIONS: Administration of rocuronium at dose of 0.25 to 0.5 mg/kg IM is a safe and effective adjunct to clinical proceedings or pre-anesthetics in P. expansa. Because rocuronium does not provide any analgesic or sedative effects, the duration of neuromuscular blockade without anesthesia should be minimized to avoid undue stress.

OBJETIVO: Determinar se o rocurônio promove imobilização segura e de curta duração em Podocnemis expansa. MÉTODOS: Vinte P. expansa com média de peso 1,59 ± 0,28 kg, foram submetidas a dois protocolos: G1 recebeu rocurônio 0,25 mg/kg IM e neostigmina 0,07 mg/kg IM enquanto G2 rocurônio 0,50 mg/kg IM e neostigmina 0,07 mg/kg IM, aplicados no membro torácico esquerdo e direito, respectivamente. Observaram-se os parâmetros anestésicos: freqüência respiratória e cardíaca, reflexo de endireitamento, relaxamento do esfíncter da cloaca, reflexo palpebral e pupilar, facilidade de manipulação, relaxamento muscular, locomoção, resposta aos estímulos dolorosos no membro torácico direito, nos membros pelvinos e na cauda, temperatura e umidade ambiental. RESULTADOS: Não foram encontradas diferenças estatísticas entre as doses para a maioria dos parâmetros e o G1 foi tão eficiente quanto o G2. Um bloqueio neuromuscular consistente foi observado aos 12 ± 4,21 minutos tanto no G1 como no G2. A recuperação de todos os animais ocorreu em até 150 minutos. CONCLUSÕES: Administração de rocurônio nas doses 0,25 e 0,50 mg/kg IM é segura e efetiva para os procedimentos clínicos ou pré-anestésicos em P. expansa. Como o rocurônio não produz efeitos sedativos ou analgésicos, a duração do bloqueio neuromuscular sem anestesia deverá ser minimizado para evitar estresse.

Pharmacological restraint of captivity giant Amazonian turtle Podocnemis expansa (Testudines, Podocnemididae) with xylazine and propofol.

PURPOSE: Identify a technique to induce brief sedation and hypnosis in Podocnemis expansa. METHODS: Twenty commercially bred P. expansa, weighing on average 1.2 +/- 0.24 kg, were subjected to two protocols: G1 was given 1.5 mg/kg IM of xylazine and 5 mg/kg IV of propofol, while G2 received 1.5 mg/kg IM of xylazine and 10 mg/kg IV of propofol. The drugs were applied, respectively, in the left thoracic member and in the cervical vertebral sinus. Assessments were made of the anesthetic parameters of locomotion, muscle relaxation, response to pain stimuli in the right thoracic members, pelvic members and tail, easy handling and heartbeat, as well as ambient temperature and glycemic level. RESULTS: A consistent hypnotic effect was recorded 49.6 +/- 22.1 seconds in G2 and after 58.2 +/- 55.1 in G1. All the animals of G2 recovered in 198 minutes, and those of G1 in 156 minutes. CONCLUSION: The hypnosis achieved with these associations was satisfactory, and G1 was as efficient as G2, allowing for the pharmacological restraint for the collection of biological samples, physical examinations and minor surgeries on these species.

Pharmacological restraint of captivity giant Amazonian turtle Podocnemis expansa (Testudines, Podocnemididae) with xylazine and propofol / Contenção farmacológica de tartaruga da Amazônia Podocnemis expansa (Testudines, Podocnemididae) de cativeiro com xilazina e propofol

PURPOSE: Identify a technique to induce brief sedation and hypnosis in Podocnemis expansa. METHODS: Twenty commercially bred P. expansa, weighing on average 1.2 ± 0.24 kg, were subjected to two protocols: G1 was given 1.5 mg/kg IM of xylazine and 5 mg/kg IV of propofol, while G2 received 1.5 mg/kg IM of xylazine and 10 mg/kg IV of propofol. The drugs were applied, respectively, in the left thoracic member and in the cervical vertebral sinus. Assessments were made of the anesthetic parameters of locomotion, muscle relaxation, response to pain stimuli in the right thoracic members, pelvic members and tail, easy handling and heartbeat, as well as ambient temperature and glycemic level. RESULTS: A consistent hypnotic effect was recorded 49.6 ± 22.1 seconds in G2 and after 58.2 ± 55.1 in G1. All the animals of G2 recovered in 198 minutes, and those of G1 in 156 minutes. CONCLUSION: The hypnosis achieved with these associations was satisfactory, and G1 was as efficient as G2, allowing for the pharmacological restraint for the collection of biological samples, physical examinations and minor surgeries on these species.

OBJETIVO: Identificar uma técnica para se induzir sedação e hipnose em Podocnemis expansa. MÉTODOS: Vinte Podocnemis expansa de criatório comercial, com média de peso 1,2 ± 0,24 kg, foram submetidas a dois protocolos: G1 recebeu xilazina 1,5 mg/kg IM e propofol 5 mg/kg IV e o G2 xilazina 1,5 mg/kg IM e propofol 10 mg/kg IV. As drogas foram aplicadas no membro torácico esquerdo e no seio vertebral cervical, respectivamente. Observaram-se os parâmetros anestésicos: locomoção, relaxamento muscular, resposta aos estímulos dolorosos no membro torácico direito, nos membros pelvinos e na cauda, facilidade de manipulação e freqüência cardíaca, além da temperatura ambiental e glicemia. RESULTADOS: Um efeito hipnótico consistente foi observado aos 49,6 ± 22,1 segundos no G2 e 58,2 ± 55,1 segundos no G1. A recuperação de todos os animais do G2 ocorreu em 198 minutos, e em 156 minutos no G1. CONCLUSÃO: A hipnose obtida por essas associações foi satisfatória e o Grupo 1 foi tão eficiente quanto o Grupo 2, o que possibilita a contenção farmacológica para coleta de amostras biológicas, exames físicos e realização de pequenas cirurgias nesta espécie.