Use of reverse transcriptase polymerase chain reaction (RT-PCR) in molecular screening of Newcastle disease virus in poultry and free-living bird populations

The aim of this study was to evaluate a simple molecular method of reverse transcriptase polymerase chain reaction (RT-PCR) to differentiate Newcastle disease virus strains according to their pathogenicity, in order to use it in molecular screening of Newcastle disease virus in poultry and free-living bird populations. Specific primers were developed to differentiate LaSota-LS-(vaccine strain) and Sao Joao do Meriti-SJM-strain (highly pathogenic strain). Chickens and pigeons were experimentally vaccinated/infected for an in vivo study to determine virus shedding in feces. Validation of sensitivity and specificity of the primers (SJM and LS) by experimental models used in the present study and results obtained in the molecular analysis of the primers by BLAST made it possible to generalize results. The development of primers that differentiate the level of pathogenicity of NDV stains is very important, mainly in countries where real-time RT-PCR is still not used as a routine test. These primers were able to determine the presence of the agent and to differentiate it according to its pathogenicity. © 2012 Springer Science+Business Media B.V.

Protection levels of vaccinated pigeons (Columba livia) against a highly pathogenic newcastle disease virus strain

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Spatial variation in post-dispersal seed removal in an Atlantic forest: Effects of habitat, location and guilds of seed predators

Studies of post-dispersal seed removal in the Neotropics have rarely examined the magnitude of seed removal by different types of granivores. The relative impact of invertebrates, small rodents, and birds on seed removal was investigated in a 2,178 ha Atlantic forest fragment in southeastern Brazil. We used popcorn kernels (Zea mays-Poaceae) to investigate seed removal in a series of selective exclosure treatments in a replicated, paired design experiment that included forest understory, gaps, and forest edge sites. We recorded the vegetation around the experimental seed stations in detail in order to evaluate the influence of microhabitat traits on seed removal. Vertebrate granivores (rodents and birds) were surveyed to determine whether granivore abundance was correlated with seed removal levels. Seed removal varied spatially and in unpredictable ways at the study site. Seed encounter and seed use varied with treatments, but not with habitat type. However, seed removal by invertebrates was negatively correlated with gap-related traits, which suggested an avoidance of large gaps by granivorous ants. The abundance of small mammals was remarkably low, but granivorous birds (tinamous and doves) were abundant at the study site. Birds were the main seed consumers in open treatments, but there was no correlation between local granivorous bird abundance and seed removal. These results emphasize the stochastic spatial pattern of seed removal, and, contrary to previous studies, highlight the importance of birds as seed predators in forest habitats. (c) 2007 Elsevier Masson SAS. All rights reserved.

Desenvolvimento e aplicação de métodos sorológicos e moleculares para o diagnóstico da doença de Newcastle em pombos comuns (Columba livia)

Pós-graduação em Microbiologia Agropecuária - FCAV

Caracterização morfológica e fenotípica de leveduras do gênero Cryptococcus com ênfase na pesquisa de Cryptococcus neoormans isoladas de amostras de água de mar e areia de três ambientes costeiros do estado de São Paulo: Canal de São Sebastião, Baixada Santista e Ubatuba

The yeast Cryptococcus neoformans is the etiologic agent of cryptococcosis, an infectious cosmopolitan disease that affects humans. Although rare, this disease is potentially fatal, especially for immunocompromised hosts. This pathogen is frequently isolated from excrements of pigeons and parrots, with many environmental sources such as birds, pigeon droppings, eucalyptus leaves, decaying trees, towers, churches and places of storage of grain (the port area). The isolation of this microorganism has been obtained also from the aquatic environment. The identification of environmental sources is needed to protect human health, especially susceptible populations such as immunocompromised. Therefore, this study investigated the presence of Cryptococcus neoformans in yeast isolates obtained from samples of sea water and sand from three regions of São Paulo: São Sebastião Channel, Santos and Ubatuba. Isolates were analyzed according to micro-and macroscopic characteristics and biochemical tests: microculture, urease, ink nankin, auxanograma, zymogram and phenol. We analyzed 199 isolates, 175 of which had features suggestive for Cryptococcus spp. in microculture. All these 175 isolates were sown in the Christensen urea middle to verify the production of urease and submitted to the technique nankin ink to visualize the capsule. Of these, only 24 were selected for the next test that was the auxanograma (assimilation of carbohydrate and nitrogen). Of the 24, 10 were tested in zymograms (fermented sugar), from which 5 were selected for the phenoloxidase test in medium containing dopamine. None of the 5 isolates tested had black or brown color characteristic of Cryptococcus neoformans. According to these tests, we arrived at 5 isolates identified to the genus Cryptococcus, but not the neoformans specie

Experimental infection of Newcastle disease virus in pigeons (Columba livia): Humoral antibody response, contact transmission and viral genome shedding

The aim of this study was to evaluate the humoral antibody response, the genome viral excretion and the contact transmission of pathogenic chicken origin Newcastle disease virus (NDV) from experimentally infected pigeons (Columba livia) to in-contact pigeon. The antibody response to infection was assessed by the hemagglutination inhibition (HI) test and the genome viral excretion was detected by RT-PCR. Viral strain induced high antibody levels, both in inoculated and in sentinel birds. The pathogenic viral strain for chickens was unable to produce clinical signs of the disease in experimentally infected pigeons, although it induced the Immoral antibody response and produced NDV genome shedding. NDV genome was detected intermittently throughout the experimental period, from 5 days post-infection (dpi) to 24 dpi. Therefore, viral genome shedding occurred for 20 days. The viral genome was detected in all birds, between I I and 13 dpi. Furthermore, the high infectivity of the virus was confirmed, as all non-inoculated sentinel pigeons showed antibody levels as high as those of inoculated birds. (C) 2007 Elsevier B.V. All rights reserved.

PREVALENCE of SALMONELLA SPP. ANTIBODIES TO TOXOPLASMA GONDII, and NEWCASTLE DISEASE VIRUS IN FERAL PIGEONS (COLUMBA LIVIA) IN THE CITY of JABOTICABAL, BRAZIL

The rock pigeon (Columba livia) may serve as a reservoir for several pathogenic agents that can be transmitted to poultry, wildlife, domesticated pets, and/or humans via excreta, secretions, or dust from feathers. In addition, ingestion of infected pigeons by wild and domestic animals can also transmit these pathogenic agents. The health status of 126 free-living pigeons in an urban area was evaluated by microbiologic culture for Salmonella and serologic testing for the presence of antibodies for Toxoplasma gondii and for Newcastle disease virus (NDV) from 120 and 109 pigeons, respectively. After drawing blood, the birds were euthanized, and fragments of the liver, spleen, lungs, and gonads, and feces were cultured for Salmonella spp. Salmonella spp. was isolated from 10 birds (7.94%), of which 8 were Salmonella typhimurium, one was Salmonella enterica subsp. enterica serotype 4,12 and one was Salmonella enterica subsp. enterica serotype 4,12,i. Six of 109 pigeons (5.50%) were positive for NDV antibodies when using the hemagglutination inhibition test. Toxoplasma gondii antibodies were detected by immunofluorescence in one of 120 sera tested (0.83%). The results indicate that feral rock pigeons were exposed to NDV and T gondii, although the exposure was low. In addition, these birds had Salmonella spp. and could disseminate this pathogen in the environment.

Chlamydophila psittaci and Toxoplasma gondii infection in pigeons (Columba livia) from São Paulo State, Brazil

Pigeons (Columba livia) cohabit with humans in urban and rural areas, representing a public health problem since microorganisms are transmitted through the inhalation of dust from their dry feces (chlamydiosis) and through ingestion of their undercooked or poorly refrigerated meat (toxoplasmosis). This study aimed to evaluate the presence of Chlamydophila psittaci and Toxoplasma gondii in pigeons from four cities in São Paulo State, Brazil. C psittaci was evaluated through hemi-nested polymerase chain reaction (hnPCR) using cloacal and tracheal swabs, whereas T. gondii specific antibodies were assessed by means of modified agglutination test (MAT), mouse brain and muscle bioassay, and polymerase chain reaction (PCR). To confirm the infection in mice, T. gondii antibodies were assayed by using indirect fluorescent antibody test (IFAT). Considering C. psittaci, 40/238 (16.8%; 95%CI 12.6-22.1%) samples were positive according to hnPCR, especially for the cities of São Paulo (42.5%) and Bauru (35%). As regards T. gondii, 12/238 (5%; 95%CI 2.9-8.6%) serum samples were positive according to MAT. of these, five samples had titer equal to 1:8; six samples, 1:16; and one sample, 1:32. Bioassay, IFAT and PCR were negative for mouse toxoplasmosis. The absence of T. gondii antibodies suggests that pigeons may be infected with a low concentration of the agent, not detected by the antigen test. Thus, C. psittaci represents an actual problem concerning bird health. (C) 2010 Elsevier B.V. All rights reserved.

Pigeons (Columba livia) are a suitable experimental model for Neospora caninum infection in birds

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Diet of Eared Doves (Zenaida auriculata, Aves, Columbidae) in a sugar-cane colony in South-eastern Brazil

Agricultores no médio Vale do Paranapanema têm relatado problemas com bandos de pombas (Zenaida auriculata) que se alimentam de cotilédones de soja na época do plantio. Na região do município de Tarumã, SP, essas aves se reproduzem em uma colônia situada em um canavial, e sua dieta é composta de 70% do peso seco por 4 grãos cultivados (em ordem de importância: milho, trigo, arroz e soja). As sementes de três invasoras (Euphorbia heterophylla, Brachiaria plantaginea e Commelina benghalensis) são importantes. Essa informação sugere que as pombas se adaptaram particularmente bem à paisagem criada pelas práticas agrícolas da região, aproveitando vários alimentos oferecidos.

Frugivory on Margaritaria nobilis L.f. (Euphorbiaceae): poor investment and mimetism.

Dehiscent fruits of Euphorbiaceae usually have two stages of seed dispersal, autochory followed by myrmecochory. Two stages of Margaritaria nobilis seed dispersal were described, the first stage autochoric followed by ornithocoric. Their dehiscent fruits are green and after they detached from the tree crown and fall on the ground, they open and expose blue metallic cocas. We studied the seed dispersal system of Margaritaria nobilis in a semi-deciduous forest in Brazil. In 80 h of focal observations, we recorded only 12 visits of frugivores, however the thrush Turdus leucomelas was the only frugivore that swallowed the fruits on the tree crown. Pitylus fuliginosus (Fringilidae) and Pionus maximiliani (Psittacidae) were mainly pulp eaters, dropping the seeds below the tree. on the forest floor, after fruits dehiscence, jays (Cyanocorax chrysops), guans (Penelope superciliaris), doves (Geotrygon montana) and collared-peccaries (Pecari tajacu) were observed eating the blue diaspores of M. nobilis. Experiments in captivity showed that scaly-headed parrots (Pionus maximiliani), toco toucans (Ramphastos toco), jays (Cyanochorax chrysops), and guans (Penelope superciliaris) consumed the fruits and did not prey on the seeds before consumption. The seeds collected from the feces did not germinate in spite of the high viability. The two stages of seed dispersal in M. nobilis resembles the dispersal strategies of some mimetic species. However M. nobilis seeds are associated with an endocarp, it showed low investment in nutrients, and consistent with this hypothesis, M. nobilis shared important characteristics with mimetic fruits, such as bright color display, long seed dormancy and protection by secondary compounds.

CHOLECYSTOKININ-LIKE IMMUNOREACTIVE RETINAL GANGLION-CELLS PROJECT TO THE VENTRAL LATERAL GENICULATE-NUCLEUS IN PIGEONS

A subpopulation of retinal ganglion cells projecting to the pigeon ventral lateral geniculate nucleus was shown to contain cholecystokinin-like immunoreactivity. These ganglion cells were mainly distributed in the peripheral retina, and their somata sizes were medium to large (14-23-mu-m). Taken together with previous findings, these results indicate that the retinal input to the ventral geniculate is chemically heterogeneous.

EFFECTS OF MASSIVE TELENCEPHALIC LESIONS ON THE ORGANIZATION OF BEHAVIOR IN PIGEONS (COLUMBA-LIVIA)

The effects of massive lesions of the telencephalon on the behavioral repertoire of captive pigeons were investigated. The behavior of four birds with an intact telencephalon was compared to that of four birds submitted to ablation of telencephalic structures. Behavioral audio recording was done according to previously defined categories in three daily sessions for fifteen days. Experimental birds were followed up for one month. Immediately after the lesion, operated birds showed decreases in the occurrence of different behaviors such as coordinated movements, feeding, interaction, preening, maintenance, exploration and vocalization and increases in locomotion and discrete movements of the body when compared to the control birds (P < 0.05). Recovery of exploration (P < 0.05), feeding and localization was observed during the follow-up period. These data are interpreted as suggestive of a functional role of the telencephalon in the organization of behavior and a long-term recovery of behavior after detelencephalation.

A liquid-phase-blocking concanavalin A enzyme-linked immunosorbent assay for the detection of antibodies against Newcastle disease virus in serum of free-ranging pigeons

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Genetic characterization of Toxoplasma gondii isolates from eared doves (Zenaida auriculata) in Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)