This study reports avian malaria caused by Plasmodium relictum in Magellanic Penguins (Spheniscus magellanicus) from São Paulo Zoo. The disease was highly infective among the birds and was clinically characterized by its acute course and high mortality. The penguins of São Paulo Zoo were housed for at least 2 years without malaria; however, they had always been maintained in an enclosure protected from mosquito exposure during the night period. When they presented pododermatitis, they were freed at night for a short period. São Paulo Zoo is located in one of the last forest remnants of the city, an area of original Atlantic forest. In the winter, the space destined for Zoo birds is shared with migratory species. Hence the possibility exists that the disease was transmitted to the penguins by mosquitoes that had previously bitten infected wild birds. Avian malaria parasites are transmitted mainly by mosquitoes of the genera Aedes and Culex, common vectors in the Atlantic forest. In this study, one Culex (Cux.) sp. was found, infected with P. relictum. There are diverse problems in housing distinct species of animals in captivity, principally when occupying the same enclosure, since it facilitates the transmission of diseases with indirect cycles, as is the case of Plasmodium spp., because certain species that cause discrete infections in some bird species can become a serious danger for others, especially penguins, which do not possess natural resistance. Thus, serious implications exist for periodically testing and administrating malaria therapy in captive penguins potentially exposed to mosquitoes during the night period, as well as other captive birds from São Paulo Zoo.

São Paulo, a densely inhabited state in southeast Brazil that contains the fourth most populated city in the world, recently experienced its largest yellow fever virus (YFV) outbreak in decades. YFV does not normally circulate extensively in São Paulo, so most people were unvaccinated when the outbreak began. Surveillance in non-human primates (NHPs) is important for determining the magnitude and geographic extent of an epizootic, thereby helping to evaluate the risk of YFV spillover to humans. Data from infected NHPs can give more accurate insights into YFV spread than when using data from human cases alone. To contextualise human cases, identify epizootic foci and uncover the rate and direction of YFV spread in São Paulo, we generated and analysed virus genomic data and epizootic case data from NHPs in São Paulo. We report the occurrence of three spatiotemporally distinct phases of the outbreak in São Paulo prior to February 2018. We generated 51 new virus genomes from YFV positive cases identified in 23 different municipalities in São Paulo, mostly sampled from NHPs between October 2016 and January 2018. Although we observe substantial heterogeneity in lineage dispersal velocities between phylogenetic branches, continuous phylogeographic analyses of generated YFV genomes suggest that YFV

Yellow Fever (YF) is a severe disease caused by Yellow Fever Virus (YFV), endemic in some parts of Africa and America. In Brazil, YFV is maintained by a sylvatic transmission cycle involving non-human primates (NHP) and forest canopy-dwelling mosquitoes, mainly Haemagogus-spp and Sabethes-spp. Beginning in 2016, Brazil faced one of the largest Yellow Fever (YF) outbreaks in recent decades, mainly in the southeastern region. In São Paulo city, YFV was detected in October 2017 in Aloutta monkeys in an Atlantic Forest area. From 542 NHP, a total of 162 NHP were YFV positive by RT-qPCR and/or immunohistochemistry, being 22 Callithrix-spp. most from urban areas. Entomological collections executed did not detect the presence of strictly sylvatic mosquitoes. Three mosquito pools were positive for YFV, 2 Haemagogus leucocelaenus, and 1 Aedes scapularis. In summary, YFV in the São Paulo urban area was detected mainly in resident marmosets, and synanthropic mosquitoes were likely involved in viral transmission.

RESUMO ABSTRACT Introduction:The objectives for this study were to measure the diversity of Culicidae species, describe their abundance and seasonal variation in São José do Rio Preto, SP, and discuss the risk of arbovirus infections. Methods: The collection of larval and adult mosquitoes was conducted monthly from 2006 to 2007 in an urban area and four sections of forested land. In the urban area, larvae were collected from sites where oviposition by Culex mosquitoes was most likely to occur. At two of the four sites in the forested land, adult mosquitoes were collected with the use of CDC traps at night, and a Nasci aspirator was used in the daytime at the two other collection sites. Results: In the urban area, 34 Culicidae species were identified out of a total sample of 8,683 specimens; of these specimens, 80.7% were Culex quinquefasciatus, 9.6% were Culex coronator, 3.2% were Aedes albopictus, and 1.1% were Ochlerotatus fluviatilis. The abundance of Cx. quinquefasciatus larvae was negatively related to rainfal. In the woods, 2,268 mosquitoes were collected, representing 10 genera and 46 species. The most abundant mosquito species were Aedeomyia squamipennis, Culex coronator, Culex (Mel.) Melanoconion section, Culex declarator, Ochlerotatus scapularis, Anopheles triannulatus, Culex bidens/interfor and Culex habilitator/pseudojhantinosoma. Conclusions: The abundance of Cx. quinquefasciatus in the urban area and the presence of other Culicidae species in urban areas and forested land point to the possibility of the transmission of West Nile virus and other arbovirus infections in São José do Rio Preto and other cities. Thus, the enacting of measures aimed at the surveillance of these arbovirus infections is essential.

-In 2000, an outbreak of sylvatic yellow fever possibly occurred in gallery forests of the Grande river in the Paraná basin in the northwestern region of São Paulo state. The aim of this study was to obtain information on the bionomics of Haemagogus and other mosquitoes inside tree holes in that area. Eighteen open tree holes were sampled for immature specimens. Adults were collected twice a month in the forest in Santa Albertina county from July 2000 to June 2001. The seasonal frequency of fourth instars was obtained by the Williams geometric mean (Mw), while the adult frequency was estimated either by hourly arithmetic or the Williams' means. Cole's index was applied to evaluate larval inter-specifi c associations. Among the ten mosquito species identifi ed, the most abundant was Aedes terrens Walker followed by Sabethes tridentatus Cerqueira and Haemagogus janthinomys Dyar. Larval and adult abundance of these species was higher in summer than in winter. Although larval abundance of Hg. janthinomys peaked in the rainy season, correlation with rainfall was not signifi cant. Six groups of larval associations were distinguished, one of which the most positively stable. The Hg. janthinomys and Ae. terrens association was signifi cant, and Limatus durhamii Theobald was the species with most negative associations.

This study compares the distribution of anopheline mosquitoes in a malaria-endemic municipality (MAL) and a malaria-free municipality (FREE) in an area of the Atlantic Forest in Brazil. Simultaneous quarterly nightly captures were made in three locations in each municipality. One Shannon light trap (Shannon light traps were home made according to specifications published in Am. J. Trop. Med. 1939; 19: 131-140) (SLT) and five CDC light traps (a kind of automatic trap fed by batteries of 12 V and 7 amp/h, with dry ice as a source of CO2; John W. Hock Company, Gainesville, FL) (CLT) (two in the canopy and three at ground level) were operated from 6:00 p.m. to 6:00 a.m. More specimens were captured in MAL (362 in SLTs and 126 in CLTs) than in FREE (66 in SLTs and 59 in CLTs). For the SLTs, Simpson's dominance index was similar in MAL and FREE (D = 0.15 versus D = 0.203, P > 0.7), but Shannon's diversity index was higher for MAL = 1.969 versus H = 1.641, P < 0.01). For the CLTs, Simpson's dominance index was higher in MAL (D = 0.416 versus 0.2688, P < 0.001), and the Shannon diversity index was higher in FREE (H = 1.5222 versus H = 1.115, P < 0.01). In SLTs, Anopheles (Kerteszia) cruzii s.l. frequencies were higher in MAL (chi2 = 23.39; P = 0.000001). In CLTs, An. cruzii s.l. was present in all strata in MAL but only in the canopy inside the forest in FREE (17 specimens). An. cruzii s.l. represented a higher proportion of anophelines in MAL (chi2 = 31; P < 0.000001). The factors that differed in these two areas were anopheline species density and An. cruzii s.l. abundance and distribution.

BACKGROUNDIn southeastern Brazil, autochthonous cases of malaria can be found near Atlantic Forest fragments. Because the transmission cycle has not been completely clarified, the behaviour of the possible vectors in those regions must be observed. A study concerning the entomological aspects and natural infection of anophelines (Diptera: Culicidae) captured in the municipalities of the mountainous region of Espírito Santo state was performed in 2004 and 2005. Similarly, between 2014 and 2015, 12 monthly collections were performed at the same area of the study mentioned above.METHODSCenter for Disease Control (CDC) light traps with CO2 were set in open areas, at the edge and inside of the forest (canopy and ground), whereas Shannon traps were set on the edge.FINDINGSA total of 1,414 anophelines were collected from 13 species. Anopheles (Kerteszia) cruzii Dyar and Knab remained the most frequently captured species in the CDC traps set in the forest canopy, as well as being the vector with the highest prevalence of Plasmodium vivax/simium infection, according to molecular polymerase chain reaction techniques.CONCLUSIONS P. vivax/simium was found only in abdomens of the mosquitoes of the subgenus Nyssorhynchus, weakening the hypothesis that this subgenus also plays a role in malaria transmission in this specific region.

Culex is the largest subgenus within the genus Culex that includes important vectors of diseases. The correct identification of mosquitoes is critical for effective control strategies. Wing geometric morphometrics (WGM) has been used to identify mosquito species alongside traditional identification methods. Here, WGM was used for eleven Culex species from São Paulo, Brazil, and one from Esquel, Argentina. Adult mosquitoes were collected using CDC (Centers for Disease Control) traps, morphologically identified and analyzed by WGM. The canonical variate analysis (CVA) was performed and a Neighbor-joining (NJ) tree was constructed to illustrate the patterns of species segregation. A cross-validated reclassification test was also carried out. From 110 comparisons in the cross-validated reclassification test, 87 yielded values higher than 70%, with 13 comparisons yielding 100% reclassification scores. Culexquinquefasciatus yielded the highest reclassification scores among the analyzed species, corroborating with the results obtained by the CVA, in which Cx. quinquefasciatus was the most distinct species. The high values obtained at the cross-validated reclassification test and in the NJ analysis as well as the segregation observed at the CVA made it possible to distinguish among Culex species with high degrees of confidence, suggesting that WGM is a reliable tool to identify Culex species of the subgenus Culex.