Multi-locus sequence typing and phylogenetics of Cryptococcus neoformans AD hybrids.

Hybrid AD strains of the human pathogenic Cryptococcus neoformans species complex have been reported from many parts of the world. However, their origin, diversity, and evolution are incompletely understood. In this study, we analyzed 102 AD hybrid strains representing 21 countries on five continents. For each strain, we obtained its mating type and its allelic sequences at each of the seven loci that have been used for genotyping haploid serotypes A and D strains of the species complex by the Cryptococcus research community. Our results showed that most AD hybrids exhibited loss of heterozygosity at one or more of the seven analyzed loci. Phylogenetic and population genetic analyses of the allelic sequences revealed multiple origins of the hybrids within each continent, dating back to one million years ago in Africa and up to the present in other continents. We found evidence for clonal reproduction and long-distance dispersal of these hybrids in nature. Comparisons with the global haploid serotypes A and D strains identified new alleles and new haploid multi-locus genotypes in AD hybrids, consistent with the presence of yet-to-be discovered genetic diversity in haploid populations of this species complex in nature. Together, our results indicate that AD hybrids can be effectively genotyped using the same multi-locus sequencing type approach as that established for serotypes A and D strains. Our comparisons of the AD hybrids among each other as well as with the global haploid serotypes A and D strains revealed novel genetic diversity as well as evidence for multiple origins and dynamic evolution of these hybrids in nature.

Cardiac Lesions of Natural and Experimental Infection by Parrot Bornaviruses.

Lymphoplasmacytic inflammation associated with bornavirus N protein occurs in the epicardial ganglia, myocardium and endocardium of birds diagnosed with proventricular dilatation disease (PDD). These pathological findings suggest that sudden death in psittacine birds might stem from cardiac compromise due to parrot bornavirus (PaBV) infection. Therefore, we investigated cardiac lesions in cases of PDD, searching databases from 1988 to 2019, and reviewed three experimental studies of PaBV infection. Fifty cases of PDD in birds infected naturally with PaBV and 27 cases of PDD in birds infected experimentally with PaBV (all having descriptions of inflammatory cardiac lesions) were reviewed. For each case, five regions of the heart were evaluated by light microscopy and immunohistochemistry (IHC). These regions were the epicardial ganglia/nerves, the endocardium, the myocardium, the Purkinje fibres and the great vessels. Sudden death was documented in 17/50 naturally infected cases, while 23/50 had digestive signs, and only 12/50 had neurological signs. Grossly, only five naturally-infected and five experimentally-infected cases had cardiomegaly or hydropericardium. Epicardial ganglioneuritis was the most consistent microscopical finding in natural (46/50) and experimental cases (26/27), followed by myocarditis (34/50) for naturally-infected and endocarditis for experimentally-infected birds (6/27). PaBV-2 antigen was detected most frequently by IHC in the epicardial ganglia (54/77) compared with the other tissues. This retrospective study demonstrates the presence of PaBV protein and inflammation in the heart of birds infected with PaBV and suggests a link between PaBV and cardiac disease and sudden death in psittacine birds.

Isolation of Cryptococcus gattii molecular type VGIII, from Corymbia ficifolia detritus in Colombia.

An environmental sampling survey was carried out in different areas of Bogotá, Colombia, to obtain isolates of members of the Cryptococcus neoformans/C. gattii species complex from Corymbia ficifolia trees. During a 6-month period in 2007, 128 samples consisting of bark, soil around trunk bases, detritus, seeds and flowers were collected from 91 trees and processed according to standard procedures. The molecular type was determined using URA5 restriction fragment length polymorphism (RFLP) analysis and the mating type was established by PCR using specific primers for Mfalpha and Mfa C. gattii was isolated from 15 of the 128 (11.7%) samples, of which three (20%) were recovered from the red flower extract and the remaining 12 from C. ficifolia detritus. URA5 RFLP analysis revealed that all 15 isolates belonged to the molecular type VGIII and mating type specific PCR revealed that all were mating type a. The isolation of C. gattii from C. ficifolia represents an important finding since this is the first report revealing C. ficifolia as a habitat for C. gattii and adds additional information to the ever growing spectrum of tree species from which C. gattii can be recovered.

Experiencia del Hospital Shriners para Niños Lisiadios, Unidad de México, en alargamientos óseos de extremidades inferiores con el método de Wagner / Experience of the Shriners Hospital for Carippled Children of Mexico in lower extremities bone lengthening by the Wagner technique

Se efectuaron 82 alargamientos con la técnica de Wagner en 33 pacientes que presentaban una discrepancia mínima de 30 milímetros y máxima de 150 milímetros, con una ganancia mínima de 10 milímetros y máxima de 67. Se presentaron múltiples complicaciones, las más frecuentes, infección del trayecto de los clavos, retardo en la consolidación y contractura en flexión de las partes blandas.