Marcela Uhart

Avian influenza virus isolated in wild waterfowl in Argentina: evidence of a potentially unique phylogenetic lineage in South America.

Avian influenza (AI) viruses have been sporadically isolated in South America. The most recent reports are from an outbreak in commercial poultry in Chile in 2002 and its putative ancestor from a wild bird in Bolivia in 2001. Extensive surveillance in wild birds was carried out in Argentina during 2006-2007. Using RRT-PCR, 12 AI positive detections were made from cloacal swabs. One of those positive samples yielded an AI virus isolated from a wild kelp gull (Larus dominicanus) captured in the South Atlantic coastline of Argentina. Further characterization by nucleotide sequencing reveals that it belongs to the H13N9 subtype. Phylogenetic analysis of the 8 viral genes suggests that the 6 internal genes are related to the isolates from Chile and Bolivia. The analysis also indicates that a cluster of phylogenetically related AI viruses from South America may have evolved independently, with minimal gene exchange, from influenza viruses in other latitudes. The data produced from our investigations are valuable contributions to the study of AI viruses in South America.

Worldwide Phylogenetic Relationship of Avian Poxviruses

ABSTRACT Poxvirus infections have been found in 230 species of wild and domestic birds worldwide in both terrestrial and marine environments. This ubiquity raises the question of how infection has been transmitted and globally dispersed. We present a comprehensive global phylogeny of 111 novel poxvirus isolates in addition to all available sequences from GenBank. Phylogenetic analysis of the Avipoxvirus genus has traditionally relied on one gene region (4b core protein). In this study we expanded the analyses to include a second locus (DNA polymerase gene), allowing for a more robust phylogenetic framework, finer genetic resolution within specific groups, and the detection of potential recombination. Our phylogenetic results reveal several major features of avipoxvirus evolution and ecology and propose an updated avipoxvirus taxonomy, including three novel subclades. The characterization of poxviruses from 57 species of birds in this study extends the current knowledge of their host range and provides the first evidence of the phylogenetic effect of genetic recombination of avipoxviruses. The repeated occurrence of avian family or order-specific grouping within certain clades (e.g., starling poxvirus, falcon poxvirus, raptor poxvirus, etc.) indicates a marked role of host adaptation, while the sharing of poxvirus species within prey-predator systems emphasizes the capacity for cross-species infection and limited host adaptation. Our study provides a broad and comprehensive phylogenetic analysis of the Avipoxvirus genus, an ecologically and environmentally important viral group, to formulate a genome sequencing strategy that will clarify avipoxvirus taxonomy.

Hematology, plasma biochemistry, and serosurvey for selected infectious agents in southern giant petrels from Patagonia, Argentina.

In conjunction with reproductive and feeding ecology studies on southern giant petrels (SGP, Macronectes giganteus) blood samples were collected for baseline health evaluations. Twenty-five adult SGP from a breeding colony in Chubut, Argentina, were sampled during two consecutive breeding seasons, 1999–2000 (n=15) and 2000–01 (n=10). Values for hematology, plasma biochemistry, and minerals are described for 20 birds in apparent good physical condition. A serologic survey of exposure to selected infectious agents was also conducted on all 25 birds sampled. Southern giant petrels were serologically negative for evidence of exposure to infectious laryngotracheitis virus, avian encephalomyelitis virus, avian influenza virus, avian reovirus, infectious bursal disease virus, infectious bronchitis virus, paramyxovirus 1, 2, and 3 virus, Chlamydophila, and Aspergillus. Antibodies to avian adenovirus were found in 14% of SGP during the first sampling season, and 60% in the second year. Additionally, all birds were negative for antibodies to Salmonella pullorum at the first sampling date, but 90% had low titers the following breeding season. This study contributes to understanding the health status of South Atlantic seabirds and to establishment of baseline information for SGP. Long-term monitoring of pelagic predator-scavenger seabirds such as SGP should be established for the surveillance of marine ecosystem health.

Health Evaluation of Pampas Deer (Ozotoceros bezoarticus celer) at Campos del Tuyú Wildlife Reserve, Argentina

Samples from 14 free-ranging pampas deer (Ozotoceros bezoarticus celer) were collected in 1995 and 1998, at Campos del Tuyú Wildlife Reserve, Buenos Aires, Argentina. Hematology, serum chemistries, minerals and metals, and fecal parasites were analyzed. In addition, fecal ova and parasites were evaluated seasonally during 1998–2000. Serology for infectious diseases included bluetongue, brucellosis, bovine respiratory syncytial virus infection, bovine viral diarrhea/mucosal disease, infectious bovine rhinotracheitis, Johnes disease (paratuberculosis), foot and mouth disease (FMD), leptospirosis (eight serovars), epizootic hemorrhagic disease, and parainfluenza-3 (PI-3). Three (21%) pampas deer had antibodies to Leptospira spp. and six (43%) to PI-3 virus. Serologic results for all other infectious agents were negative. Domestic cattle (n=27) included in this study for comparison had antibodies to Leptospira, infectious bovine rhinotracheitis virus, bovine viral diarrhea virus, and PI-3 virus (74–100% of tested animals) and one animal (4%) to Brucella sp. All cattle had antibodies to FMD virus attributable to vaccination. This study provides the first data on the health status of the southernmost subspecies of pampas deer.

Disease Survey of Free-ranging Grey Brocket Deer (Mazama gouazoubira) in the Gran Chaco, Bolivia

Samples from 17 free-ranging hunter-killed grey brocket deer (Mazama gouazoubira) in the Gran Chaco, Bolivia, were collected during June–August 1999. All 17 deer appeared to be in good condition at the time of death. Gross necropsies were performed, serum was collected for serologic evaluation of selected infectious disease agents, and feces and ectoparasites were collected for evaluation of internal and external parasites. Serologic tests were positive for antibodies against bovine respiratory syncytial virus and four Leptospira interrogans serovars, with questionable results for epizootic hemorrhagic disease virus serotypes 1 and 2. No antibodies were detected to Anaplasma marginale, Babesia bigemina, Babesia bovis, Babesia odocoilei, bluetongue virus (serotypes 2, 10, 11, 13, and 17), bovine viral diarrhea virus, Brucella abortus, foot-and-mouth disease virus, infectious bovine rhinotracheitis virus, Mycobacterium avium subsp. paratuberculosis, and parainfluenza-3 virus. Sixty-four percent (7/11) of the deer had en-doparasites. Amblyomma spp. ticks were found on seven deer, flies of the family Hippoboscidae on six deer, and lice on six deer.

Recent and Chronic Exposure of Wild Ducks to Lead in Human-modified Wetlands in Santa Fe Province, Argentina

Poisoning of waterfowl due to ingestion of lead pellets is a worldwide problem in areas that are subject to hunting. No studies have assessed exposure of waterbirds to this heavy metal in Argentina, in spite of intense hunting activity, and the fact that only lead ammunition is commercially available. The objective of this study was to evaluate duck exposure to lead by examining gizzard and bone samples collected from 30 wild ducks, 16 Rosy-billed Pochard (Netta peposaca), and 14 Fulvous Whistling-Duck (Dendrocygna bicolor), provided by hunters in northern Santa Fe Province, Argentina, in July 2007. Radiographs, followed by dissection of the gizzards, showed that 31% of the Rosy-billed Pochards and 29% of the Fulvous Whistling-Ducks had ingested lead pellets (between one and four per animal). Lead in bone was found at concentrations associated with detrimental health effects. In spite of the small number of samples in this project, these results indicate high levels of lead exposure (both recent and chronic) in these species. This is the first report of a problem in Argentina that could represent a threat to the health and conservation of native aquatic species, their predators, and the wetlands they inhabit.

Sex determination of adult Rock Shags by molecular sexing and morphometric parameters

Abstract Male and female Rock Shags (Phalacrocorax magellanicus) are not obviously sexually dimorphic in plumage or size and are thus difficult to distinguish in the field. We evaluated the utility of two different DNA-based techniques for sexing adult Rock Shags. We found that the primer set 2550F/2718R (originally tested in three individuals of P. carbo), with minor differences in the forward primer, provided a consistent and simple sexing method for Rock Shags. Moreover, we obtained three reliable discriminant functions for sexing adults from three different colony sites between 42° to 47°S in coastal Patagonia, Argentina. Discriminant analysis of five external characters of adult birds indicated that head, bill, and wing lengths were the most accurate variables for use in a discriminant function model, predicting the sex of 80–86% of the birds. Males were significantly larger than females for all body measurements except for bill depth. Rock Shags showed less marked sexual dimorphism than other phalacrocoraciid species.

Numerical and Spatial Responses of Geoffroy's Cat (Oncifelis geoffroyi) to Prey Decline in Argentina

Abstract We examined the numerical and spatial responses of Geoffroys cats (Oncifelis geoffroyi) to a prey decline in central Argentina between April 2002 and November 2003. The 2nd year of the study coincided with a severe drought. Relative abundance of brown hares (Lepus europaeus) declined from 5.6 individuals/10 km during the predrought period to about 0.6 individuals/10 km during the drought. Small-rodent biomass also showed the lowest level for the study area during the drought of 2002–2003 (134.5 g/ha). During the predrought and drought periods, 3 male and 1 female, and 1 male and 9 female Geoffroys cats, respectively, were radiotagged and monitored. Home ranges for males of the predrought period averaged 202.8 ha ± 156.8 SD and that of the single female was 27.3 ha. During the drought period, 4 females occupied an average home range of 254.9 ± 254.1 ha, and the home-range size of the single predrought female increased by a factor of 2. No obvious change in mean daily distance traveled between the 2 periods was observed. Geoffroys cats predominantly used habitats of dense cover during the predrought period, but they became more habitat generalists during the drought. Recruitment of juveniles was only recorded during the predrought period, and all monitored Geoffroys cats dispersed or died of starvation after the prey decline. Consequently, density of Geoffroys cats dropped from 2.9 individuals/10 km2 before the drought to 0.3 individuals/10 km2, probably because of food scarcity. This is the 1st study to examine the spatial ecology of a small wild cat species under nutritional (energetic) stress in South America.

Avian Pox in Magellanic Penguins (Spheniscus magellanicus)

Avian pox is an enveloped double-stranded DNA virus that is mechanically transmitted via arthropod vectors or mucosal membrane contact with infectious particles or birds. Magellanic Penguins (Spheniscus magellanicus) from two colonies (Punta Tombo and Cabo Dos Bahías) in Argentina showed sporadic, nonepidemic signs of avian pox during five and two of 29 breeding seasons (1982– 2010), respectively. In Magellanic Penguins, avian pox expresses externally as wart-like lesions around the beak, flippers, cloaca, feet, and eyes. Fleas (Parapsyllus longicornis) are the most likely arthropod vectors at these colonies. Three chicks with cutaneous pox-like lesions were positive for Avipoxvirus and revealed phylogenetic proximity with an Avipoxvirus found in Black-browed Albatross (Thalassarche melanophrys) from the Falkland Islands in 1987. This proximity suggests a long-term circulation of seabird Avipoxviruses in the southwest Atlantic. Avian pox outbreaks in these colonies primarily affected chicks, often resulted in death, and were not associated with handling, rainfall, or temperature.

Phylogenetic Analysis of H6 Influenza Viruses Isolated from Rosy-Billed Pochards (Netta peposaca) in Argentina Reveals the Presence of Different HA Gene Clusters

ABSTRACT Until recently, influenza A viruses from wild waterfowl in South America were rarely isolated and/or characterized. To explore the ecology of influenza A viruses in this region, a long-term surveillance program was established in 2006 for resident and migratory water birds in Argentina. We report the characterization of 5 avian influenza viruses of the H6 hemagglutinin (HA) subtype isolated from rosy-billed pochards (Netta peposaca). Three of these viruses were paired to an N2 NA subtype, while the other two were of the N8 subtype. Genetic and phylogenetic analyses of the internal gene segments revealed a close relationship with influenza viruses from South America, forming a unique clade and supporting the notion of independent evolution from influenza A viruses in other latitudes. The presence of NS alleles A and B was also identified. The HA and NA genes formed unique clades separate from North American and Eurasian viruses, with the exception of the HA gene of one isolate, which was more closely related to the North American lineage, suggesting possible interactions between viruses of North American and South American lineages. Animal studies suggested that these Argentine H6 viruses could replicate and transmit inefficiently in chickens, indicating limited adaptation to poultry. Our results highlight the importance of continued influenza virus surveillance in wild birds of South America, especially considering the unique evolution of these viruses.