First evidence of the emergence of novel putative infectious bronchitis virus genotypes in Cuba

The emergence of new infectious bronchitis virus (IBV) genotypes or serotypes along with the poor cross-protection observed among IBV serotypes have complicated the avian infectious bronchitis (IB) control programs in different geographic regions. In Cuba, the lack of genetic information regarding IBV and the increasing epidemiological importance of this virus in Cuban chicken flocks demand further characterization of IBV isolates. In the present work, studies of genetic diversity and phylogenetic relationships among recent IBV isolates from Cuban chicken flocks showing respiratory disorders were performed. Two putative genotypes genetically different to the Massachusetts genotype H120 strain used in the Cuban vaccination program were found in the flocks assessed. In addition, a potential nephropathogenic IBV isolate was found by first time in Cuba. (C) 2012 Elsevier Ltd. All rights reserved.

WATER VAPOR CONDUCTANCE OF THE LOWER CRETACEOUS DINOSAURIAN EGGS FROM SANAGASTA, LA RIOJA, ARGENTINA: PALEOBIOLOGICAL AND PALEOECOLOGICAL IMPLICATIONS FOR SOUTH AMERICAN FAVEOLOOLITHID AND MEGALOOLITHID EGGS

The water vapor conductance (G(H20)) of the neosauropod eggs from the Lower Cretaceous Sanagasta nesting site in La Rioja Province, Argentina, was examined and compared with other Cretaceous Argentinean oological material. The 2900 mgH(2)O/day.Torr G(H2O) of the Sanagasta eggshells confirms an extremely moist nesting environment and supports field observations of dug-out nests in a geothermal setting. The observed thinning of the outer eggshell surface during incubation increases gas conductance and concomitantly decreases eggshell mechanical resistance during the late ontogenetic stages, thus facilitating embryonic development and hatching. The Sanagasta and Entre Rios Province faveoloolithid eggs display the highest and comparable 61120 values and share several morphological and diagenetic characters, indicating comparable nesting strategy in geothermal settings. However, the faveoloolithid Yamintie and La Pampa Province specimens cluster together with lower G(H20) values closer to the megaloolithid eggs. The Gnu) of the megaloolithid egg Megaloolithus patagonicus was reconsidered and new results are now congruent with other reported megaloolithid GH2O values. Additionally, we hypothesize that V-shaped pore canals of M. patagonicus, which upper sections reach only the top third or half eggshell thickness and, a wider section in the middle would not compromise the overall egg mechanical resistance like vertical pores connecting directly the outer to the inner eggshell surfaces. Such pore spatial arrangement and geometry would enhance, as the eggshell thins during incubation, a greater G(H2O), G(O2) and G(CO2) and facilitate embryonic development in high moisture nesting contents. Overall, data suggests that neosauropod nesting and brooding behaviors were dependent on elevated moisture nesting environments.

Purificação e caracterização da VDAC de mitocôndrias corticais aviares: identificação de modificações pós-traducionais nas porinas neuronais murinas e aviares

A VDAC é uma porina presente na MME cuja função é crucial no metabolismo energético, sobrevivência e morte celular. A caracterização da VDAC torna-se importante para a compreensão das inter-relações da mitocôndria com os diferentes componentes citosólicos, tais como a HK. A ligação HK-VDAC favorece a utilização do ATP intramitocondrial em células neuronais, a HK cerebral pode interagir de formas diferentes com a VDAC, o que resulta em diferentes sítios de ligação (sítios A e B). Os variados papéis metabólicos das isoformas da VDAC podem ser explicados pela presença de alterações pós-traducionais. No presente trabalho purificamos a VDAC1 mitocondrial neuronal proveniente de cérebro aviar. Paralelamente, comprovamos que a presença de múltiplas formas das VDACs 1 e 2 em cérebros murino e aviar, seja devida à presença de modificações pós-traducionais, nomeadamente a fosforilação. A proteína isolada apresentou peso molecular de 30KDa. Quando submetida à eletroforese e posteriormente à coloração para a identificação de fosfoproteínas, a mesma mostrou-se desfosforilada. O conhecimento da presença, ou ausência de fosforilação das VDACs, reside na importância de estabelecer-se as bases moleculares ligadas à existência de sítios A e B nas mitocôndrias neuronais.

Molecular Surveillance of the Newcastle Disease Virus in Domestic and Wild Birds on the North Eastern Coast and Amazon Biome of Brazil

Brazil is one of the world's largest countries with a rich diversity of wildlife, including resident and migratory wild birds, which may be natural reservoirs of the Newcastle disease virus (NDV). Because Brazil is a major global exporter of chicken meat, the emergence of such a disease may have a huge negative impact not only on the economy due to trade restrictions and embargoes, but also on the quality of life of the population. Samples were collected from 1,022 asymptomatic domestic and wild birds from the Brazilian coast and the Amazon region using tracheal/cloacal swabs and tested by RT-qPCR. The results showed 7 (0.7%) birds were positive for NDV. The positive samples were then isolated in embryonated chicken eggs and their matrix protein genes were partially sequenced, revealing a low-pathogenicity NDV. This study confirms the maintenance of the velogenic-NDV free status of Brazil.

Hematologic and Plasma Biochemical Values of Hyacinth Macaws (Anodorhynchus hyacinthinus)

The hyacinth macaw (Anodorhyncus hyacinthinus), considered the largest psittacine bird species in the world, is an endangered species, with a remaining population of approximately 6500 birds in the wild. To establish hematologic and plasma biochemical reference ranges and to verify differences related to sex, samples from 29 hyacinth macaws (14 males. 15 females) were obtained from birds apprehended from illegal wildlife trade and subsequently housed at the Sorocaba Zoo, Brazil. No significant differences in hematologic or plasma biochemical values were found between females and males. Compared with published reference values, differences were found in mean concentrations of total red blood cell count, corpuscular volume, corpuscular hemoglobin level, total white blood cell count, aspartate aminotransferase level, creatine kinase concentration, alkaline phosphatase concentration, and phosphorus level. Baseline hematologic and plasma biochemical ranges were established, which may be useful as reference values for clinicians working with this endangered species in captivity or rehabilitation centers.

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/mm(3), 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/mm(3), leukocytes 11.9 +/- 4.5 x 10(3) cells/mm(3), heterophils 6284 +/- 2715 cells/mm(3), lymphocytes 5473 +/- 2408 cells/mm(3), monocytes 113 +/- 162 cells/mm(3), eosinophils 10 +/- 42 cells/mm(3), basophils 27 +/- 64 cells/mm(3). 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 < 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.

A NEW GENUS AND SPECIES OF PHILOPTERIDAE (PHTHIRAPTERA: ISCHNOCERA) FROM THE TRUMPETERS (AVES: GRUIFORMES: PSOPHIIDAE)

A new chewing louse genus and species belonging to the Philopteridae, namely, Palmaellus inespectatus n. gen., n. sp., is described. The new genus is distinguished from the other ischnoceran genera hitherto described by its peculiar characters of the dorsal anterior head plate with 2 postero-lateral projections, pterothorax and abdomen with scarce chaetotaxy, male genitalia with simple mesomere and paramere lacking inner digitiform projection, and the genital region of female with postero-vulvar plates bearing setae. It is a parasite of the trumpeters, an avian family endemic to South America's Amazon Basin.

The legs: a key to bird evolutionary success

Birds are the most diverse and largest group of extant tetrapods. They show marked variability, yet much of this variation is superficial and due to feather and bill color and shape. Under the feathers, the skeleto-muscular system is rather constant throughout the bird group. The adaptation to flight is the explanation for this uniformity. The more obvious morphological adaptations for flight are the wings, but the trunk is always rigid, the tail is short and the neck is flexible, since all these features are correlated with flying behaviour. Unrelated to the exigencies of flight, the legs always have three long bones, and all the birds walk on their toes. This leg structure is a striking plesiomorphic feature that was already present in related dinosaurs. The multi-purpose potential of the legs is the result of the skeletal architecture of a body with three segmented flexed legs. This configuration provides mechanical properties that allow the use of the legs as propulsive, paddling, foraging or grooming tools. It is the association of diverse modes of locomotion-walking, running, hopping, flying and swimming-that have enabled the birds to colonize almost all the environments on Earth.

Life-cycle and host preference of Amblyomma ovale (Acari: Ixodidae) under laboratory conditions

This study evaluated for the first time the life cycle of Amblyomma ovale in the laboratory. For this purpose, larvae and nymphs were exposed to Gallus gallus (chickens), Cavia porcellus (guinea pigs), Rattus norvegicus (wistar rats), Oryctolagus cuniculus (domestic rabbits), Calomys callosus (vesper mouse), and Didelphis albiventris (white-eared opossum). Nymphs were also exposed to Nectomys squamipes (South American water rat). Adult ticks were fed on dogs. The life-cycle of A. ovale in laboratory could be completed in an average period of ca. 190 days, considering prefeeding periods of 30 days for each of the parasitic stages. Vesper mice were the most suitable host for A. ovale larvae, whereas water rats were the most suitable host for A. ovale nymphs. Our results, coupled with literature data, strongly indicate that small rodents have an important role in the life history of A. ovale. Chickens (the only avian host used in the present study) showed to be moderately suitable hosts for subadult A. ovale ticks, indicating that wild birds might have a secondary role in the life history of A. ovale. Domestic dogs showed to be highly suitable for the adult stage of A. ovale, in agreement with literature data that indicate that the domestic dog is currently one of the most important hosts of A. ovale adult ticks in Latin America.

Heat stress impairs performance and induces intestinal inflammation in broiler chickens infected with Salmonella Enteritidis

Stressful situations reduce the welfare, production indices and immune status of chickens. Salmonella spp. are a major zoonotic pathogens that annually cause over 1 billion infections worldwide. We therefore designed the current experiment to analyse the effects of 31 +/- 1 degrees C heat stress (HS) (from 35 to 41 days) on performance parameters, Salmonella invasion and small intestine integrity in broiler chickens infected with Salmonella Enteritidis. We observed that HS decreased body weight gain and feed intake. However, feed conversion was only increased when HS was combined with Salmonella Enteritidis infection. In addition, we observed an increase in serum corticosterone levels in all of the birds that were subjected to HS, showing a hypothalamus-pituitary-adrenal axis activation. Furthermore, mild acute multifocal lymphoplasmacytic enteritis, characterized by foci of heterophil infiltration in the duodenum, jejunum and ileum, was observed in the HS group. In contrast, similar but more evident enteritis was noted in the heat-stressed and Salmonella-infected group. In this group, moderate enteritis was observed in all parts of the small intestine. Lastly, we observed an increase in Salmonella counts in the spleens of the stressed and Salmonella-infected chickens. The combination of HS and Salmonella Enteritidis infection may therefore disrupt the intestinal barrier, which would allow pathogenic bacteria to migrate through the intestinal mucosa to the spleen and generate an inflammatory infiltrate in the gut, decreasing performance parameters.

Multiple independent origins of mitochondrial control region duplications in the order Psittaciformes

mitochondrial genomes are generally thought to be under selection for compactness, due to their small size, consistent gene content, and a lack of introns or intergenic spacers. As more animal mitochondrial genomes are fully sequenced, rearrangements and partial duplications are being identified with increasing frequency, particularly in birds (Class Ayes). In this study, we investigate the evolutionary history of mitochondrial control region states within the avian order Psittaciformes (parrots and cockatoos). To this aim, we reconstructed a comprehensive multi-locus phylogeny of parrots, used PCR of three diagnostic fragments to classify the mitochondrial control region state as single or duplicated, and mapped these states onto the phylogeny. We further sequenced 44 selected species to validate these inferences of control region state. Ancestral state reconstruction using a range of weighting schemes identified six independent origins of mitochondrial control region duplications within Psittaciformes. Analysis of sequence data showed that varying levels of mitochondrial gene and tRNA homology and degradation were present within a given clade exhibiting duplications. Levels of divergence between control regions within an individual varied from 0-10.9% with the differences occurring mainly between 51 and 225 nucleotides 3' of the goose hairpin in domain I. Further investigations into the fates of duplicated mitochondrial genes, the potential costs and benefits of having a second control region, and the complex relationship between evolutionary rates, selection, and time since duplication are needed to fully explain these patterns in the mitochondrial genome. (C) 2012 Elsevier Inc. All rights reserved.

Real-time PCR assay targeting the actin gene for the detection of Cryptosporidium parvum in calf fecal samples

Cryptosporidium parvum infection is very important with respect to public health, owing to foodborne and waterborne outbreaks and gastrointestinal illness in immunocompetent and immunocompromised persons. In cattle, infection with this species manifests either as a subclinical disease or with diarrheal illness, which occurs more often in the presence of other infectious agents than when alone. The aim of this study was to develop a real-time polymerase chain reaction (PCR) assay for the detection of C. parvum in calf fecal samples and to compare the results of this assay with those of the method routinely used for the diagnosis of Cryptosporidium spp., nested PCR targeting the 18S rRNA gene. Two hundred and nine fecal samples from calves ranging in age from 1 day to 6 months were examined using real-time PCR specific for the actin gene of C. parvum and by a nested PCR targeting the 18S rRNA gene of Cryptosporidium spp. Using real-time PCR detection, 73.2% (153 out of 209) of the samples were positive for C. parvum, while 56.5% (118 out of 209) of the samples were positive for Cryptosporidium spp. when the nested PCR amplification method was used for the detection. The analytical sensitivity of the real-time PCR was approximately one C. parvum oocyst. There was no significant nonspecific DNA amplification of any of the following species and genotype: Cryptosporidium andersoni, Cryptosporidium baileyi, Cryptosporidium bovis, Cryptosporidium canis, Cryptosporidium galli, Cryptosporidium ryanae, Cryptosporidium serpentis, or avian genotype II. Thus, we conclude that real-time PCR targeting the actin gene is a sensitive and specific method for the detection of C. parvum in calf fecal samples.

The effect of premedication with ketamine, alone or with diazepam, on anaesthesia with sevoflurane in parrots (Amazona aestiva)

Abstract Background Premedication is rarely used in avian species. The aim of this study was to evaluate the effect of premedication on the quality of sevoflurane induction and anaesthesia in parrots. We hypothesised that premedication would facilitate handling and decrease the minimum anaesthetic dose (MAD). Thirty-six adult parrots were randomly distributed in three groups: group S (n = 12) was premedicated with NaCl 0.9%; group KS (n = 12) was premedicated with 10 mg.kg-1 ketamine; and group KDS (n = 12) was premedicated with 10 mg.kg-1 ketamine and 0.5 mg.kg-1 diazepam, delivered intramuscularly. After induction using 4.5% sevoflurane introduced through a facemask, the MAD was determined for each animal. The heart rate (HR), respiratory rate (RR), systolic arterial blood pressure (SAP), and cloacal temperature (CT) were recorded before premedication (T0), 15 minutes after premedication (T1), and after MAD determination (T2). Arterial blood gas analyses were performed at T0 and T2. The quality of anaesthesia was evaluated using subjective scales based on animal behaviour and handling during induction, maintenance, and recovery. Statistical analyses were performed using analysis of variance or Kruskal-Wallis tests followed by Tukey’s or Dunn’s tests. Results The minimal anaesthetic doses obtained were 2.4 ± 0.37%, 1.7 ± 0.39%, and 1.3 ± 0.32% for groups S, KS, and KDS, respectively. There were no differences in HR, RR, or CT among groups, but SAP was significantly lower in group S. Sedation was observed in both the premedicated S-KS and S-KDS groups. There were no differences in the quality of intubation and recovery from anaesthesia among the three groups, although the induction time was significantly shorter in the pre-medicated groups, and the KS group showed less muscle relaxation. Conclusions Ketamine alone or the ketamine/diazepam combination decreased the MAD of sevoflurane in parrots (Amazona aestiva). Ketamine alone or in combination with diazepam promoted a good quality of sedation, which improved handling and reduced the stress of the birds. All protocols provided safe anaesthesia in this avian species.

Comparação quantitativa da comunidade de aves de um fragmento de floresta semidecidual do interior do Estado de São Paulo em intervalo de 30 anos

Few studies have evaluated long-term changes in avian abundance in forest remnants. To compare both species richness and abundance of the bird community in a forest fragment located in the municipality of Gália, state of São Paulo, southeastern Brazil, we surveyed forest birds using transect counts. We compared our results with a survey conducted 30 years earlier at the same locality and further classified bird species according to their food habits to eventually predict fluctuations of specific abundance. Although species with population declines predominated in the community, all trophic categories had species which increased their abundances. Most species prone to move around remnants decreased in abundance. We suggest that, regarding specific abundances, trophic categories may be equally affected as a result of fragmentation processes and that the forest regeneration of this remnant may have led to the loss of edge species. Species that suffered from abundance loss during this time period may become locally extinct in the near future.

Survey on Chlamydophila psittaci in captive ramphastids in São Paulo State, Brazil

Chlamydophila psittaci (C. psittaci) has been detected in 460 avian species, among them the most frequent are the Psittaciformes, Columbiformes, Anseriformes and raptors. In Brazil, the main avian species recognized as healthy carriers belong to the order Psittaciformes and Columbiformes, but very few studies have been done in other bird families. Reports of the occurrence of this disease in the clinical form are rare in the Ramphastids; consequently, they are not commonly evaluated for this agent. The present study reports the investigation of C. psittaci in 25 captive ramphastids from a zoological park in São Paulo State, Brazil. Swabs samples from the cloaca were submitted to semi-nested polymerase chain reaction (semi-nested PCR) for direct detection of the microorganism. Additionally, blood samples obtained from these birds were submitted to the Complement Fixation Test (CFT) for detection of antibodies anti-C. psittaci. The presence of C. psittaci was not detected in the cloacal swab samples tested by the PCR. Nevertheless, 16% (4/25) of the bird's sera were positive by the CFT. Among the species with positive results, there are the saffron toucanet (Pteroglossus bailloni) and black-necked-aracari (Pteroglossus aracari), two species with no descriptions of the survey of C. psittaci published in the literature. Intermittent elimination of C. psittaci is a feature of chronically infected birds; however the absence of a positive-antigen sample did not guarantee that the bird is Chlamydophila-free. The serological results obtained show that the ramphastids tested were previously exposed to the pathogen and developed immune response, but showed no clinical signs of the disease and didn't eliminate regularly the organism in their feces in the moment of the sample collection.