Comparison of Serum Thyroid Hormone Levels in Green-winged Macaws (Ara chloropterus) Using Radio and Chemiluminescent Immunoassays.

This study compared the use of radioimmunoassay (RIA) and chemiluminescent immunoassay (CLIA) to measure serum thyroid hormone levels in green-winged macaws (Ara chloropterus). The sample group comprised 20 male and 13 female (n = 33) healthy, captive green-winged macaws of reproductive age but outside of breeding season. Mean free triiodothyronine (T3), total T3, and free thyroxine (T4) measured by CLIA and RIA corresponded to 5.11 and 5.08 pmol/L (3.33 and 3.31 pg/mL), 1.59 and 1.56 nmol/L (103.5 and 101.5 ng/dL), and 8.25 and 8.82 pmol/L (0.64 and 0.69 ng/dL), respectively. Mean total T4 by RIA corresponded to 6.29 nmol/L (0.49 µg/dL). Mean free T4 and thyroid-stimulating hormone (TSH) levels differed according to immunoassay method, with higher values measured by RIA compared with CLIA. Sex influence was statistically significant in immunoassay results only on free T3 levels. Free T3 levels measured by RIA were higher than levels measured by CLIA in male birds, but this difference was not found with the female bird samples. Conversely, free T3 levels measured by CLIA were higher in the female macaws compared with male birds. Comparative analysis of thyroid hormone measurements in this study revealed that RIA and CLIA are equivalent methods to measure free T4 and total T3 levels but not TSH levels. These findings support the use of CLIA for free T4 and total T3 level determinations in green-winged macaws. However, the CLIA kit used in this study provided invalid total T4 level results for the macaws sampled. Radioimmunoassay and CLIA were equally ineffective for determining TSH levels in this species.

Fecal sex steroids and reproductive behaviors in harpy eagles (Harpia harpyja).

Aiming to improve our reproductive knowledge of large birds of prey, behavioral data and fecal steroids were assessed in captive pairs of Harpy eagles, a keystone species that is monogamous and resides in the Neotropics year-round. Adult individuals exhibited different reproductive outcomes and a breeding season extending beyond summer solstice (5-9 months) suggests that harpy eagles may not be absolutely photorefractory. Comparisons among breeding stages in males revealed that mean androgen levels in courtship were higher than in copulation and incubation, but no differences were detected in fecal progestagens or estrogens. Females had higher mean estrogen concentrations in courtship and copulation, whereas mean progestagen levels peaked during egg laying. Mean androgen concentrations were not significantly different among breeding stages in females. Assessment of six egg-lay cycles from three females demonstrated that fecal estrogens peaked predominantly between 31 and 18 days before oviposition (-31 to -18 days), and then remained low until 45 days after laying the first egg (+45 days). In contrast, fecal progestagens raised mostly between -20 and +1 day, lowering to baseline concentrations by +3 days. To our knowledge, this is the first study to describe in detail endocrine and behavioral data regarding reproduction in tropical eagles, which may serve in the future as a reference to developing breeding programs.

Microscopic and molecular identification of hemotropic mycoplasmas in South American coatis (Nasua nasua).

Hemoplasmas were detected in two apparently healthy captive South American coatis (Nasua nasua) from southern Brazil during an investigation for vector-borne pathogens. Blood was subjected to packed cell volume (PCV) determination, a commercial real-time PCR panel for the detection of Anaplasma spp., Babesia spp., Bartonella spp., Hepatozoon spp., Leishmania spp., Mycoplasma haemofelis, 'Candidatus Mycoplasma turicensis', 'Candidatus Mycoplasma haemominutum', Neorickettsia risticii, Rickettsia rickettsii and Leptospira spp., and a pan-hemoplasma conventional PCR assay. PCV was normal, but both coatis tested positive for hemoplasmas and negative for all the remaining pathogens tested. Using different techniques for microscopy (light, confocal or SEM), structures compatible with hemoplasmas were identified. Sequencing of the 16S rRNA gene identified an organism resembling Mycoplasma haemofelis and another hemotropic Mycoplasma sp., with a sequence identity of 96.8% to a Mycoplasma sp. previously detected in capybaras.

Occurrence of hemotropic mycoplasmas in non-human primates (Alouatta caraya, Sapajus nigritus and Callithrix jacchus) of southern Brazil.

Hemoplasmas, the erythrocyte-associated mycoplasmas, have been detected in several primates, causing mostly subclinical infection. This study aimed to determine the prevalence of hemoplasma infection in captive and free-ranging monkeys from southern Brazil, as well as factors and hematological abnormalities associated with infection. Blood samples from 40 non-human primates (NHP) were tested for hemoplasmas and coinfections. An overall of 10/40 (25.0%) NHP tested positive for hemoplasmas using PCR-based assays, including 9/14 (64.3%) black howler monkeys (Alouatta caraya) and 1/24 (4.2%) black-horned capuchin (Sapajus nigritus). Infection was not statistically associated with anemia, but wild-born monkeys and male black howler monkeys were more likely to be positive when compared with captive-born animals and female black howler monkeys, respectively. The sequences from the black howler monkey hemoplasma were similar (94% identity) to the squirrel monkey hemoplasma ("Candidatus Mycoplasma kahanei") and were phylogenetically located in a different cluster when compared to the human hemoplasma ("Candidatus Mycoplasma haemohominis").

Doppler ultrasonography of the pectinis oculi artery in harpy eagles (Harpia harpyja).

Twenty harpy eagles (Harpia harpyja) without systemic or ocular diseases were examined to measure blood velocity parameters of the pectinis oculi artery using Doppler ultrasonography. Pectinate artery resistive index (RI) and pulsatility index (PI) were investigated using ocular Doppler ultrasonography. The mean RI and PI values across all eyes were 0.44±0.10 and 0.62±0.20 respectively. Low RI and PI values found in the harpy eagle´s pectinis oculi artery compared with the American pekin ducks one and other tissue suggest indeed a high metabolic activity in pecten oculi and corroborates the hypothesis of a nutritional function and/or intraocular pressure regulation.

The use of altrenogest to avoid hyperestrogenism after eCG-hCG ovulation induction in southern tigrina (Leopardus guttulus).

The goal of this study was to optimize an ovulation induction protocol for use with artificial insemination in the southern tigrina (Leopardus guttulus). The specific aims were to report the efficacy of using altrenogest, an oral progestin (Regumate, MSD Animal Health, Merck Animal Health), to suppress ovarian activity and prevent follicular hyperstimulation and hyperestrogenism after the administration of exogenous eCG and hCG. To monitor ovarian responses, fecal estrogen and progestogen metabolites were quantified by enzyme immunoassay in females before and after intramuscular administration of 200-IU eCG and 150-IU hCG in two trials, 4 months apart. During the first trial, there was no use of altrenogest, only the eCG-hCG ovulation induction protocol. In the second trial, the ovulation induction protocol was preceded by the administration of oral altrenogest for 14 days (minimum of 0.192 mg per kg per day). Altrenogest administration resulted in a suppression of follicular activity in three out of six females before eCG-hCG administration on the basis of lower mean estrogen concentrations (P < 0.05). It also resulted in four out of six females presenting lower fecal estrogen metabolite concentrations (P < 0.05) after ovulation induction, and two out of six individuals showed a reduction (P < 0.05) in postovulatory fecal progestogen metabolite concentrations, all when compared to the same female's cycles without the progestin. Fecal estrogen metabolite concentrations were closer to baseline in 50% of these individuals after altrenogest and eCG-hCG treatments when compared to basal concentrations before gonadotropins without the use of altrenogest. This study reported that use of altrenogest in southern tigrina can suppress ovarian activity and avoid hyperestrogenism after administration of eCG and hCG treatment. However, not all females responded uniformly, so more studies are needed to increase the efficacy of ovulation induction for use with artificial insemination in this species.

Haematological values for captive harpy eagle (Harpia harpyja) / Valores hematológicos para águia hapia (Harpia harpyja) em cativeiro

Decreasing of harpy eagle (Harpia harpyja) populations in natural environments, mainly in non-preserved areas, makes captive population management an important contribution to genetic diversity conservation. The aim of this study is to evaluate hematological parameters for captive harpy eagles maintained at the wild animals breeding center of Itaipu Binacional, Paraná State, Brazil. Fourteen blood samples from nine harpy eagles were collected from animals of both sexes, of different ages and with no clinical signs of disease. Significant variations were found in haematological values of hematocrit, hemoglobin, mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), leukocyte, a relative number of heterophils, absolute and relative number of lymphocytes, monocytes, eosinophils, basophils and plasma protein between groups of young (less than six months old) and adult birds. Comparing males and females there was variation in the values of erythrocytes, hemoglobin, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) on heterophils, absolute and relative number of lymphocytes, eosinophils and basophils. There was also variation in the values of red blood cells, hematocrit, hemoglobin, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), leukocyte count, absolute number of lymphocytes, eosinophils and basophils among birds that study compared to another reference birds. Due to the limited information available on harpy eagle hematology, this study will be useful to the clinical assessment of birds maintained in captivity...

A diminuição de populações de harpia (Harpia harpyja) em ambientes naturais, principalmente em áreas não preservadas, torna a gestão população cativa uma contribuição importante para a conservação da diversidade genética. O objetivo deste estudo é avaliar os parâmetros hematológicos de harpias em cativeiro mantidas para os animais selvagens de reprodução centro da Itaipu Binacional, Paraná, Brasil. Quatorze amostras de sangue de nove harpias foram coletadas de animais de ambos os sexos, de diferentes idades e sem sinais clínicos da doença. Variações significativas foram encontradas nos valores hematológicos de hematócrito, hemoglobina, hemoglobina corpuscular média (HCM), concentração de hemoglobina corpuscular média (CHCM), de leucócitos, um número relativo de heterófilos, absoluta e número relativo de linfócitos, monócitos, eosinófilos, basófilos e proteínas plasmáticas entre grupos de jovens (menos de seis meses de idade) e adultos aves. Comparando homens e mulheres houve variação nos valores de eritrócitos, hemoglobina, volume corpuscular médio (VCM), hemoglobina corpuscular média (HCM) em heterófilos, o número absoluto e relativo de linfócitos, eosinófilos e basófilos. Houve também variação nos valores de células vermelhas do sangue, hematócrito, hemoglobina, volume corpuscular médio (VCM), hemoglobina corpuscular média (HCM), concentração de hemoglobina corpuscular média (CHCM), contagem de leucócitos, número absoluto de linfócitos, eosinófilos e basófilos entre aves que estudam em comparação com outras aves de referência. Devido à pouca informação disponível sobre hematologia harpia, este estudo será útil para a avaliação clínica das aves mantidas em cativeiro...

Comparison of methods for determination of nutrient digestibility of a dry kibble diet for ocelots / Comparação de métodos para estimar coeficientes de digestibilidade de uma ração comercial de gatos-domésticos para jaguatiricas

In this experiment, methods of total fecal collection (TFC) and internal markers (acid-insoluble ash - AIA, crude fiber - CF, and acid-detergent fiber - ADF) were compared for determination of the coefficients of apparent digestibility (CAD) for dry matter (DM), crude protein (CP), ether extract (EE), nitrogen-free extracts (NFE), and gross energy (GE) of commercial feline dry kibble for ocelots (Leopardus pardalis). Six adult animals, weighing 12.45±1.37 kg, gradually received experimental kibble in their usual diet until the beginning of the experiment and were submitted to an adaptation period ten days prior to the collection period. CAD obtained by TFC, AIA, CF, and ADF were, respectively, 73.7, 76.83, 62.01, and 46.03% for dry matter; 81.9, 84.8, 75.8, and 63.8% for crude protein; 85, 86.7, 78.5, and 69.1% for ether extract; 78.52, 79.55, 69.11, and 53.04% for nitrogen-free extracts; and 80.5, 82.2, 71.4, and 58.4% for gross energy. The AIA method showed to be efficient in determining coefficients of apparent digestibility and may contribute to investigations on the digestibility of diets for wild felines. In comparison to the items of ocelot's usual diet, the kibble used in this paper provided an adequate nutritional supply with reduced daily costs per animal.

Neste experimento foram comparados os métodos de coleta total de fezes (CT) e de indicadores internos (cinza insolúvel em ácido - CIA, fibra bruta - FB e fibra em detergente ácido - FDA) na determinação dos coeficientes de digestibilidade aparente (CDA) da matéria seca (MS), proteína bruta (PB), extrato etéreo (EE), extrativo não nitrogenado (ENN) e energia bruta (EB) de uma ração comercial de gatos-domésticos para jaguatiricas (Leopardus pardalis). Seis animais adultos com peso de 12,45 ± 1,37kg receberam gradativamente a ração experimental na dieta habitual até o início do experimento e foram submetidos a um período de adaptação de 10 dias anteriores ao período de coleta. Os CDA obtidos pela CT, CIA, FB e FDA foram, respectivamente de 73,70; 76,83; 62,01 e 46,03% para matéria seca, 81,9; 84,8; 75,8 e 63,8% para proteína bruta, 85,0; 86,7; 78,5 e 69,1% para extrato etéreo, 78,52; 79,55; 69,11 e 53,04% para extrativo não nitrogenado e de 80,5; 82,2; 71,4 e 58,4% para energia bruta. O método de CIA mostrou-se eficiente na determinação dos coeficientes de digestibilidade aparente e pode contribuir com as investigações sobre a digestibilidade em dietas com felídeos selvagens. A ração utilizada permitiu aos animais um adequado aporte nutricional e apresentou menores custos/animal/dia em comparação aos itens que compunham a dieta habitual.

Hemotropic mycoplasma in a free-ranging black howler monkey (Alouatta caraya) in Brazil.

Hemotropic mycoplasmas are bacteria that infect erythrocytes and cause subclinical infections to life-threatening disease. We describe hemotropic mycoplasma infection in a free-ranging black howler monkey (Alouatta caraya). This is the first molecular detection of a hemotropic mycoplasma in a nonhuman primate from Brazil.

Serologic survey for Leptospira spp. in captive neotropical felids in Foz do Iguaçu, Paraná, Brazil.

Leptospirosis is a bacterial zoonosis of worldwide distribution and is endemic in tropical countries, where rodents and other wild mammals are abundant and may act as reservoirs. Leptospirosis has become a concern in captive wild animals, due mostly to their exposure to contaminated urine or environment. Although domestic cats (Felis catus) have been reported refractory to leptospirosis, serology and disease in captive wild felids is still unclear. In this study 57 adult, clinically healthy felids, including 1 Geoffroy's cat (Leopardus geoffroyi), 3 jaguarundis (Puma yagouaroundi), 17 margays (Leopardus wiedii), 22 little spotted cats (Leopardus tigrinus), and 14 ocelots (Leopardus pardalis) kept in captivity at the Sanctuary at the Itaipu Binacional hydroelectric power plant (Bela Vista Biological Sanctuary), Foz do Iguacu City, Paraná State, Brazil, were serologically surveyed for the presence of antibodies against 28 serovars of Leptospira spp. by microagglutination test (MAT). Two animals (3.5%) were seropositive: one male ocelot to the serovar Cynopteri (titer 100) and one female margay to Autumnalis (100) and Butembo (200). The captive-born, 5-yr-old ocelot had been solitary housed in an individual cage. The approximately 21-yr-old wild-caught margay was also kept individually. None of the tested animals showed signs ofleptospirosis. During a study conducted 4 yr previously in the same facility, this particular margay also tested positive for the same two serovars, among others. The present study indicates that the felids tested for Leptospira spp. by MAT were exposed to serovars, but did not demonstrate clinical signs of disease. Comparison with a previous study suggests that serovar titers may vary over time and that leptospirosis dynamics remains unclear in wild felids.

Anti-Rickettsia spp. antibodies in free-ranging and captive capybaras from southern Brazil / Anticorpos anti-Rickettsia spp. em capivaras de vida livre e de cativeiro no Sul do Brasil

Capybaras (Hydrochaeris hydrochaeris) are among the main hosts of Amblyomma spp. ticks, which is able to transmit Rickettsia species to human beings and animals. Since they are often infested with potential vector ticks, capybaras may be used as sentinels for rickettsiosis, such as the Brazilian Spotted Fever. The aim of the present study was to determine the prevalence of antibodies against Rickettsia spp. using the indirect immunofluorescence assay (IFA) in 21 free-ranging and 10 captive animals from the Zoological Park of the 'Bela Vista Biological Sanctuary' (BVBS), Itaipu Binational, Foz do Iguaçu, Southern Brazil. Antigens of six rickettsial species already identified in Brazil (Rickettsia rickettsii, R. parkeri, R. bellii, R. rhipicephali, R. amblyommii and R. felis) were used for IFA. Ticks from each capybara were collected for posterior taxonomic identification. A total of 19 (61.3 percent) samples reacted to at least one of tested species. Seropositivity was found in 14 (45.2 percent), 12 (38.7 percent), 5 (16.1 percent), 4 (12.9 percent), 3 (9.7 percent) and 3 (9.7 percent) animals for R. rickettsii, R. bellii, R. parkeri, R. amblyommii, R. felis and R. rhipicephali, respectively. Two captive capybaras presented suggestive titers of R. rickettsii infection and one sample showed homologous reaction to R. parkeri. Only one free-ranging capybara presented evidence R. bellii infection. Ticks collected on capybaras were identified as Amblyomma dubitatum e Amblyomma sp. Results evidenced the rickettsial circulation in the area, suggesting a potential role of capybaras on bacterial life cycle.

As capivaras (Hydrochaeris hydrochaeris) estão entre os principais hospedeiros do carrapato Amblyomma spp., o qual pode transmitir algumas espécies de riquétsias para seres humanos e animais. Como são frequentemente infestadas por carrapatos vetores potenciais, as capivaras podem ser usadas como sentinelas para riquetsioses, como a Febre Maculosa Brasileira. O objetivo do presente estudo foi determinar a soroprevalência de Rickettsia spp. por meio da reação de Imunofluorescência Indireta (RIFI) em 21 capivaras de vida livre e 10 capivaras de cativeiro do Zoológico do Refúgio Biológico Bela Vista, Itaipu Binacional, Foz do Iguaçu, Brasil. Antígenos de seis espécies de riquétsias já identi[1]icadas no Brasil (Rickettsia rickettsii, R. parkeri, R. bellii, R. rhipicephali, R. amblyommii e R. felis) foram utilizados para a RIFI. Carrapatos de cada capivara foram coletados para posterior identi[1]icação taxonômica. Um total de 19 (61,3 por cento) amostras reagiu a pelo menos uma das espécies testadas. Foi encontrada soropositividade em 14 (45,2 por cento), 12 (38,7 por cento), 5 (16,1 por cento), 4 (12,9 por cento), 3 (9,7 por cento) e 3 (9,7 por cento) animais para R. rickettsii, R. bellii, R. parkeri, R. amblyommii, R. felis e R. rhipicephali, respectivamente. Duas capivaras de cativeiro apresentaram títulos sugestivos de infecção por R. rickettsii e uma amostra apresentou reação homóloga frente à R. parkeri. Apenas uma capivara de vida livre apresentou evidência de infecção por R. bellii. Os carrapatos coletados sobre as capivaras foram identificados como Amblyomma dubitatum e Amblyomma sp. Os resultados evidenciam a circulação de riquétsias na região, sugerindo uma potencial participação da capivara no ciclo de vida desta bactéria.

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 (Refúgio 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.

Detection of a novel hemoplasma based on 16S rRNA gene DNA in captive and free-ranging capybaras (Hydrochaeris hydrochaeris).

Two different species of hemoplasmas, Mycoplasma coccoides and M. haemomuris, are known to infect small rodents such as mice and rats. However, there are no previous reports of hemoplasma infection in capybara (Hydrochaeris hydrochaeris). The aim of our study was to determine whether these hemoplasmas might infect capybaras from Southern Brazil. Blood samples from 31 animals: 10 captive and 21 free-ranging capybaras were collected and packed cell volume and total plasma protein were measured. DNA was extracted and PCR assays for M. coccoides and M. haemomuris were performed. Using the M. coccoides-PCR assay 64% of the capybaras were positive, 80% free-ranging and 30% from captive animals. The prevalence of infection between the groups was significantly different (p=0.001). Sequencing of the nearly entire 16S rRNA gene from the positive samples suggested a novel hemoplasma isolate with identity of 92% with M. coccoides and 86% with M. haemomuris. All capybara samples were negative for M. haemomuris infection. DNA of a housekeeping gene was successfully amplified from all samples. This is the first evidence of a hemoplasma infection in capybaras.