Epidemiology of neglected tropical diseases in transplant recipients: review of the literature and experience of a Brazilian HSCT center

The rising success rate of solid organ (SOT) and haematopoietic stem cell transplantation (HSCT) and modern immunosuppression make transplants the first therapeutic option for many diseases affecting a considerable number of people worldwide. Consequently, developing countries have also grown their transplant programs and have started to face the impact of neglected tropical diseases (NTDs) in transplant recipients. We reviewed the literature data on the epidemiology of NTDs with greatest disease burden, which have affected transplant recipients in developing countries or may represent a threat to transplant recipients living in other regions. Tuberculosis, Leprosy, Chagas disease, Malaria, Leishmaniasis, Dengue, Yellow fever and Measles are the topics included in this review. In addition, we retrospectively revised the experience concerning the management of NTDs at the HSCT program of Amaral Carvalho Foundation, a public transplant program of the state of São Paulo, Brazil.

SAINT LOUIS ENCEPHALITIS VIRUS IN MATO GROSSO, CENTRAL-WESTERN BRAZIL

The dengue virus (DENV), which is frequently involved in large epidemics, and the yellow fever virus (YFV), which is responsible for sporadic sylvatic outbreaks, are considered the most important flaviviruses circulating in Brazil. Because of that, laboratorial diagnosis of acute undifferentiated febrile illness during epidemic periods is frequently directed towards these viruses, which may eventually hinder the detection of other circulating flaviviruses, including the Saint Louis encephalitis virus (SLEV), which is widely dispersed across the Americas. The aim of this study was to conduct a molecular investigation of 11 flaviviruses using 604 serum samples obtained from patients during a large dengue fever outbreak in the state of Mato Grosso (MT) between 2011 and 2012. Simultaneously, 3,433 female Culex spp. collected with Nasci aspirators in the city of Cuiabá, MT, in 2013, and allocated to 409 pools containing 1-10 mosquitoes, were also tested by multiplex semi-nested reverse transcription PCR for the same flaviviruses. SLEV was detected in three patients co-infected with DENV-4 from the cities of Cuiabá and Várzea Grande. One of them was a triple co-infection with DENV-1. None of them mentioned recent travel or access to sylvatic/rural regions, indicating that transmission might have occurred within the metropolitan area. Regarding mosquito samples, one pool containing one Culex quinquefasciatus female was positive for SLEV, with a minimum infection rate (MIR) of 0.29 per 1000 specimens of this species. Phylogenetic analysis indicates both human and mosquito SLEV cluster, with isolates from genotype V-A obtained from animals in the Amazon region, in the state of Pará. This is the first report of SLEV molecular identification in MT.

DENGUE OUTBREAK IN MATO GROSSO STATE, MIDWESTERN BRAZIL

Dengue virus (DENV) is the most frequent arbovirus worldwide. In this study, we report a large outbreak in Mato Grosso State (MT). Serum samples from 604 patients with acute febrile illness for less than five days were inoculated in C6/36 cells, then infected cells were subjected to an indirect immunofluorescence test for DENV serotypes and yellow fever virus. Serum samples were submitted to a multiplex-semi-nested-RT-PCR for 11 flaviviruses. DENV-4 was isolated in 150/604 (24.8%) and DENV-1 in 19/604 (3.1%) specimens. By RT-PCR, 331 (54.8%) samples tested positive for DENV; 321 had single infections (DENV-4 n = 305; DENV-1 n = 15; DENV-3 n = 1), nine had co-infections of DENV-1/DENV-4, and one of DENV-2/DENV-4. DENV-4 was detected in 315/331 (95.2%) positive patients from 17 municipalities, and DENV-1 in 24/331 (7.2%) patients from five cities in north-central MT and the city of Cuiaba. The incidence of infection was higher in patients aged 20-39 (142/331; 42.9%). The NS5 partial nucleotide sequence of DENV-1 was most similar to that of genotype V, DENV-2 to Southeast Asian/American, DENV-3 to genotype III, and DENV-4 to genotype II strains, considered the most frequent strains in Brazil. This outbreak coincided with the introduction of DENV-4 in the state. Cuiaba was hyperendemic for the four DENV serotypes, highlighting the necessity for arbovirus surveillance in MT.

Software for pattern recognition of the larvae of Aedes aegypti and Aedes albopictus

Software for pattern recognition of the larvae of mosquitoes Aedes aegypti and Aedes albopictus, biological vectors of dengue and yellow fever, has been developed. Rapid field identification of larva using a digital camera linked to a laptop computer equipped with this software may greatly help prevention campaigns.

Emergent arboviruses in Brazil

Brazil is a large tropical country (8,514,215km²) with 185,360,000 inhabitants. More than one third of its territory is covered by tropical forests or other natural ecosystems. These provide ideal conditions for the existence of many arboviruses, which are maintained in a large variety of zoonotic cycles. The risk that new arboviruses might emerge in Brazil is related to the existence of large, densely populated cities that are infested by mosquitoes such as Culex and the highly anthropophilic Aedes aegypti. Infected humans or animals may come into these cities from ecological-epidemiological settings where arbovirus zoonoses occur. This study analyzes the risk of emergence of the alphaviruses Mayaro, Venezuelan equine encephalitis, Eastern equine encephalitis and Chikungunya; the flaviviruses yellow fever, Rocio, Saint Louis encephalitis and West Nile; and the orthobunyavirus Oropouche.

Concurrent dengue and malaria in the Amazon region

INTRODUCTION: The Amazon region has extensive forested areas and natural ecosystems, providing favorable conditions for the existence of innumerous arboviruses. Over 200 arboviruses have been isolated in Brazil and about 40 are associated with human disease. Four out of 40 are considered to be of public health importance in Brazil: Dengue viruses (1-4), Oropouche, Mayaro and Yellow Fever. Along with these viruses, about 98% of the malaria cases are restricted to the Legal Amazon region. METHODS: This study aimed to investigate the presence of arboviruses in 111 clinical serum samples from patients living in Novo Repartimento (Pará), Plácido de Castro (Acre), Porto Velho (Rondônia) and Oiapoque (Amapá). The viral RNA was extracted and RT-PCR was performed followed by a Multiplex-Nested-PCR, using Flavivirus, Alphavirus and Orthobunyavirus generic and species-specific primers. RESULTS: Dengue virus serotype 2 was detected in two patients living in Novo Repartimento (Pará) that also presented active Plasmodium vivax infection. CONCLUSIONS: Despite scant data, this situation is likely to occur more frequently than detected in the Amazon region. Finally, it is important to remember that both diseases have similar clinical findings, thus the diagnosis could be made concomitantly for dengue and malaria in patients living or returning from areas where both diseases are endemic or during dengue outbreaks.

Atypical lymphocytosis in leptospirosis: a cohort of hospitalized cases between 1996 and 2009 in State of Rio de Janeiro, Brazil

INTRODUCTION: Leptospirosis is a zoonotic disease found in tropical and temperate countries, and its clinical diagnostic confusion with arboviruses (dengue fever, oropouche fever and yellow fever), Brazilian spotted fever, viral hepatitis and hantaviruses has been an ongoing public health concern. The aim of this observational study was to demonstrate an association between findings of atypical lymphocytosis and the progression of endemic leptospirosis. METHODS: A retrospective analysis was performed on the demographic, epidemiological, clinical and laboratory aspects of 27 human leptospirosis cases that occurred over a period of 13 years (1996-2009) with no reported epidemic outbreaks in Rio de Janeiro, Brazil. RESULTS: The overall mortality rate was 11.1% in our cohort of hospitalized cases. However, there was no mortality among patients with atypical lymphocytosis (OR = 11.1; 95% CI = 1.12-110.9; p = 0.04). Two patients who were in the septicemic phase showed signs of expansion of γδ T cell responses in peripheral blood. CONCLUSIONS: Atypical lymphocytosis may be observed in patients with leptospirosis. Our observations suggest that these atypical leukocyte subsets are associated with partial protection during the disease course of leptospirosis.

Bioactivity of plant extracts on the larval and pupal stages of Aedes aegypti (Diptera, Culicidea)

Introduction Aedes aegypti is responsible for the transmission of the dengue and yellow fever viruses. This study evaluated the effects of extracts from Cnidosculos phyllacanthus, Ricinus communis, and Coutarea hexandra on the developmental periods of A.aegypti larvae and pupae. Crude extracts of C. phyllacanthus and C. hexandra and oil from R. communis and C. phyllacanthus were used. Methods Bioassays of the larvicidal and pupicidal effects of these products at different concentrations and times of exposure were evaluated. The lethal and sublethal effects were determined using different concentrations in larvicidal tests. Mortality data were evaluated by Probit analysis to determine the LC50 and LC90 values. Results The vegetable oils from C. phyllacanthus and R. communis demonstrated greater efficiency for larval control with an LC50=0.28µl/mL and an LC90=1.48µl/mL and LC50=0.029µl/mL and a LC90=0.26µl/mL, respectively. In pupal tests toxic effects for all insects were verified after exposure to the products at significant LC50 and LC90 values for 24 and 48h. The effects of sublethal concentrations of C. phyllacanthus (oil) were more effective on the insects. Conclusions The vegetables oils from C. phyllacanthus and R. communis demonstrated greater potential from the control of different developmental periods in the life cycle of this insect.

Natural transovarial transmission of dengue virus 4 in Aedes aegypti from Cuiabá, State of Mato Grosso, Brazil

INTRODUCTION: Dengue is the most prevalent arboviral disease in tropical areas. In Mato Grosso, outbreaks are reported every year, but studies on dengue in this state are scarce. METHODS: Natural transovarial infection of Aedes aegypti by a flavivirus was investigated in the Jardim Industriário neighborhood of Cuiabá, Mato Grosso. Eggs were collected with ovitraps during the dry, intermediate, and rainy seasons of 2012. After the eggs hatched and the larvae developed to adulthood, mosquitoes (n = 758) were identified and allocated to pools of 1-10 specimens according to the collection location, sex, and climatic period. After RNA extraction, multiplex semi-nested RT-PCR was performed to detect the four dengue virus (DENV) serotypes, yellow fever virus, West Nile virus and Saint Louis encephalitis virus. RESULTS: DENV-4 was the only flavivirus detected, and it was found in 8/50 pools (16.0%). Three of the positive pools contained females, and five contained males. Their nucleotide sequences presented 96-100% similarity with DENV-4 genotype II strains from Manaus, Amazonas. The minimum infection rate was 10.5 per 1000 specimens, and the maximum likelihood estimator of the infection rate was 11.6 (95% confidence interval: 4.8; 23.3). CONCLUSIONS: This study provides the first evidence of natural transovarial infection by DENV-4 in Ae. Aegypti in Mato Grosso, suggesting that this type of infection might serve as a mechanism of virus maintenance during interepidemic periods in Cuiabá, a city where dengue epidemics are reported every year. These results emphasize the need for efficient vector population control measures to prevent arbovirus outbreaks in the state.

Arboviral diseases in the Western Brazilian Amazon: a perspective and analysis from a tertiary health & research center in Manaus, State of Amazonas

The Fundação de Medicina Tropical Dr. Heitor Vieira Dourado (FMT-HVD), located in Manaus, the capital of the State of Amazonas (Western Brazilian Amazon), is a pioneering institution in this region regarding the syndromic surveillance of acute febrile illness, including arboviral infections. Based on the data from patients at the FMT-HVD, we have detected recurrent outbreaks in Manaus by the four dengue serotypes in the past 15 years, with increasing severity of the disease. This endemicity has culminated in the simultaneous circulation of all four serotypes in 2011, the first time this has been reported in Brazil. Between 1996 and 2009, 42 cases of yellow fever (YF) were registered in the State of Amazonas, and 71.4% (30/42) were fatal. Since 2010, no cases have been reported. Because the introduction of the yellow fever virus into a large city such as Manaus, which is widely infested by Aedes mosquitoes, may pose a real risk of a yellow fever outbreak, efforts to maintain an appropriate immunization policy for the populace are critical. Manaus has also suffered silent outbreaks of Mayaro and Oropouche fevers lately, most of which were misdiagnosed as dengue fever. The tropical conditions of the State of Amazonas favor the existence of other arboviruses capable of producing human disease. Under this real threat, represented by at least 4 arboviruses producing human infections in Manaus and in other neighboring countries, it is important to develop an efficient public health surveillance strategy, including laboratories that are able to make proper diagnoses of arboviruses.

Zika virus in Brazil and the danger of infestation by Aedes (Stegomyia) mosquitoes

Abstract Zika virus, already widely distributed in Africa and Asia, was recently reported in two Northeastern Brazilian: State of Bahia and State of Rio Grande do Norte, and one Southeastern: State of São Paulo. This finding adds a potentially noxious virus to a list of several other viruses that are widely transmitted by Aedes (Stegomyia) aegypti and Aedes (Stegomyia) albopictus in Brazil. The pathology and epidemiology, including the distribution and vectors associated with Zika virus, are reviewed. This review is focused on viruses transmitted by Aedes (Stegomyia) mosquitoes, including dengue, Chikungunya, Zika, Mayaro, and yellow fever virus, to emphasize the risks of occurrence for these arboviruses in Brazil and neighboring countries. Other species of Aedes (Stegomyia) are discussed, emphasizing their involvement in arbovirus transmission and the possibility of adaptation to environments modified by human activities and introduction in Brazil.

Ixodidas brasileiros e de alguns paizes limitrophes

The knowledge of the Ixodidae becomes every day, more and more important owing to the fact of the increasing number of diseases of man and animals they can transmit. In Brasil besides transmitting treponemosis, piroplasmosis and anaplasmosis to several domestic animals, the ticks are also responsible fo the transmission of the brazilian rocky mountain spotted fever (A. cajennense and Amblyomma striatum) and they can also harbour the virus of the yellow fever and even to transmit it in laboratory experiments (A. cajennense, O. rostratus). The Brazilian fauna of ticks is a small one and has no more than 45 well-established species belonging to the genus Argas, Ornithodoros, Ixodes, Haemaphysalis, Rhipicephalus, Boophilus, Amblyomma and Spaelaeorhynchus. The genus Amblyomma is the best represented one, with 67% of all species of ticks known in Brazil. One of the most important species in the Amblyomma cajennense owing to its abundance and its wide parasitism in many vertebrates: reptiles, birds and mammals, incluing man, who is much attacked by the larva, the nymph and the adult of this species. The other ticks who attack the man are the Amblyomma brasiliense (the pecari tick), in the forests, and the Ornithodoros, especially the species. O. rostratus and brasiliensis. Other species can bite the man, but only occasionally, like Amblyomma fossum, striatum, oblongogutatum etc. Argas persicus, Rhipicephalus sanguineus and Boophilus are very important species not only as parasites but specially because they transmit several diseases to animals. Some of the ticks of the brazilian wild animals are now also parasites of the domestic ones and vice-versa. Arga persicus var. dissimilis is very common among the poultry and transmits the Treponema anserinum (gallinarum). Boophilus microplus is very abundant on our domestic and wild ruminants (Bos, Cervus, Mazama etc.) and can also ben found on horse, dogs, Felis onca, Felis concolor etc., and it transmits to cattle piroplasmosis and anaplasmosis. Rhipicephalus sanguineus (an introduced species) is now very common on the dog, over all the country. The author recommend to give popular names to some brazilian ticks in order to make them more acquainted with the non scientific people. The author gives a classification of the superfamilia Ixoidoidea and keys to the determination of the different species of brazilian ticks. He creates a new family of Nuttallielidae to the so interesting tick, described by Bedford with the name of Nuttaliella namaqua in South Africa, a new variety of Argas persicus, the Argas persicus var. dissimilis nov. var. owing to the differences on the segment and on the size and morphology of the peritrema. He describes also the female of Amblyomma fuscum Nn. A great part of the author's work deals with the biology, life conditions and parasitism of many of the brazilian ticks in accordance with his personal and from other author's researches, especially in reference to Argas persicus, Ornithodoros rostratus, O. brasiliensis, Boophilus microplus, Rhipicephalus sanguineus, Amblyomma cajennense, A. pseudoconcolor, A. auriculare, A. rotundatum (= A. agamum) etc. The author gives a detailed report upon the parthenogenesis of A. rotundatum (A. agamum) that he first described in 1912 and gives also many references to other species of brazilian ticks, to teratological forms etc. He also gives a detailed report of the geographical distribution of brazilian ticks and of the peculiar conditions of its parasitism. The last part of this article deals with references to the species of ticks of some of the South American Republics namely Argentina, Bolivia, Colombia, Paraguay and Venezuela. Amblyomma testudinis Conil, A. neumanni Ribaga 1902 (= A. furcula Dõnitz 1909) and A. parvitarsum Nn. 1899 (= A. altiplanum Dios 1917), are found only in Argentina. It is given a special bibliography dealing with the brazilian ticks and four text figures and one plate.

Uma estação biológica para o estudo dos mosquitos e dos outros animaes silvestres relacionados com a febre amarela

The outbreak of the jungle or forest yellow fever, through the adapta¬tion, quite recently of the yellow fever virus o the forest mosquitoes, brou¬ght the necessity of ecological researches on hese mosquitoes, as well as on the wild animals they bite, some of them being susceptible to the desease. This has been done by the special yellow fever Service of the State of Sao Paulo, in a special Biological Station in Perús, São Paulo, which has been built in the midst of the jungle. This station was made with plain materials, and covered with straw, but was confortable enough for the technical work, i nthe early months of 1938. During the months in which the investigations were being carried on, the following interesting results were obtained: 1. As we have already pointed out in other places, the forest mosquitoes biting us during daytime, are always new born insects, having not yet sucked blood, as it is the general rule with all mosquitoes, and therefore also, with the anopheles and stegomyia, and this explains why nobody gets malaria or yellow fever, transmitted by anofeles or by aedes aegypti during the day. We think therefore, the jungle yellow fever, got during daytime is not due to the infected jungle or forest mosquito biting, but to infection through the human skin coming into close contact with tre virus, which the forest mosquitoes lay with their dejections, on the leaves of the trees where they remain sitting du¬ring the day. 2. As it is the rule with anopheles, stegomyia and other mosquitoes, the insects once having sucked blood, take nocturnal habits and, therefore, bite us, only during the night, so it happens with the forest mosquito, and insects with developped eggs and blood in stomach have been caught within the sta¬tion house, during the night. During the day, these mosquitoes do not bite, but remain quite still on the leaves of the trees, in the damp parts of the woods. 3. Jungle or forest mosquitoes can easely bite wild animals, some with more avidity then ethers, as it has bee npointed out to the opossum (didei-phis) and other animals. They also bite birds having very thin skin and only exceptionally, cold bloods animals. 5. Is has hot been possible to ascertain how forest mosquitoes are able to live, from onde season to another, through winter, when temperature drops near and even below zero. They have not been found in holes of the terrain, of trees and of animals, as it is the rule in cold countries. During winter, in the forest, it is possible to find larvs in the holes of bambus and trees full of water. As wild animals do not harbour the yellow fever virus for a long time in their body, it is diffcult to explain how the desease lasts from one season to another. Many ecological features on the mosquito, remains yet to be explained and therefore it in necessary to go on with the investigations, in bio¬logical stations, such as that one built up in Perús, São Paulo.

Estudos dos agrupamentos vegetativos relacionados com as áreas onde foram efetuadas as pesquisas sôbre a febre amarela silvestre no Município de Passos, Estado de Minas Gerais

The work reported here was carried-out on the invitation of Dr. Henry Kumm, Director of the Rockefeller Foundation, and by appointment from Dr. Henrique Aragão, Director of the Instituto Oswaldo Cruz. It was done during the investigation of sylvan yellow fever, in June 1947, with a view to establishing the phyto-ecological conditions of the county of Passos. The pe¬riod was, however, too short for definite conclusions to be reached. Thanks are due to Dr. O. R. Causey, Chief of Research on Yellow Fever for transpor¬tation and other help. THE REGIONAL VEGETATION. Aerial photographs of the county of Passos shoto that it is covered by three great types of vegetation: Rain Forest, Secondary Pasture Land and Scrub.1 Detailed investigation, however, brings out the fact that these correspond to different seres; furthermore, each type presents not only the specific, characteristics of the biological form dominant for the climate, but also are at various stages, which express HABITATS differing from those of the normal sere. The phytogeographic survey of the region shows that most of it is now covered by secondary pasture land (disclimax) in which Melinis minutiflora, v. "fat grass" (fig. 1), predominates. The mosaic of Rain Forest and of small patches of Scrub reveals the effects of human intervention (BARRETO, H. L. de Mello 1); consequently, all the formations have to be regarded as secon¬dary, though some of them probably include relicts of the primitive climax (WARMING, E. 2). On close examination, the Scrub cannot be considered as the climax, because of the following facts: 1. In the zone of Rain-Forest stretches of forest are present in very varied topographic conditions and the reconstitution of the associations show that man has destroyed an ecological unit (fig. 2). 2. In the zone of Scrub the characteristic patches are small. The banks of rivers and brooks, the valleys and ravine and whatever the soil has retained some humidity, is being invaded fry Rain Forest, which seems to be growing under optimum conditions. The Scrub is thus limited to small belts on the calcareous mountains and on sandy soils with alcaline depths (pH abo¬ve 7) which do not retain enough moisture for the Rain Forest that is progres¬sively restricting the area occupied by Scrub. In view of the topographic and present climatic conditions the Rain Forest must consequently be regarded as the regional climax. The presence of ecologically contradictory elements and associations shows that the real problem is that of the fluctuations of the climate of Passos or even of Minas Geraes during the quaternary and recent periods (DAN-SEREAU, P. : 3), a subject on which little is known and which is tied to the evolution of the climate of Brazil (OLIVEIRA, E. : 4) . The transformation of Scrub into Rain Forest has been - observed by the author before, in other parts of Brazil (VELOSO, PL P.: 5) . It seems probable that the Rio Grande has also greatly influenced the change of the regional vegetation, by invading areas of Scrub and dislocating the limit of the Pluvial climate towards the Canastra Range, though there are remnants of Scrub (postclimax) transfor¬med into secondary open country (disclimax, fig. 5) by human devastation and the setting of fire to the land. VEGETATION GROUPS OF THE PLUVIAL TYPE. The map of the region also shows that at the present time the small patches of forest (whether devasted or intact) occupy the least accessible places, such as valleys, peaks and abrupt slopes (fig. 2). Even these are now being destroyed, so that in the near future this forested region will be en¬tirely reduced to poor pasture land unless energetic measures of conservation are undertaken in time. The Special Service for Prophylaxis against Yellow Fever installed two of their four Stations for the Capture of Mosquitos in this area, one of them at Batatal and the other at Cachoeira, which have separate formations each of them composed of several associations. Other vegetation formations were also analysed, from the synecological point of view, so as to ascertain of which degree of succession their associations belong. These phytosociological sur¬veys give an idea of the principal characteristics of each station. BATATAL FORMATION. The abrupt nature of the valley has rendered this location inappropriate for agricultural purposes since colonial times. The relict of the primitive forest climax saved by this circumstance has expanded gradually to zones whose paedologic conditions favour the eatablishment of mesophilous species. The aerial photograph shows two small stretches of forest, one apparently primi¬tive, the other composed of associations belonging to the subclimax of the subsere. CACHOEIRA FORMATION. Aerial photographs show that this station is crossed by a small river, which divides it into two separate parts. The first, which presents ecological conditions similar, though not identical to those of Batatal, is favoured by topography and apparently remains primitive forest. Though the topography of the other, on the whole, favours the establishment of groups belonging to the normal sere of the climax, is has been partly devastated recently and the aspect of the associations has been completely modified. It was is this part that the four posts for the capturing of mosquitos were set up. The first forest is favoured by deposition of organic matter, washed out from the nearby devasted areas by torrential rains, and thus provides, an appropriate HABITAT for the climax species with certain hygrophilous trends of the ecological quasiclimax type. This association seems to have reached a biological equilibrium, as the dominates. Gallesia gorarema and Cariniana legalis (fig. 10), present an optimum vitality with a vigorous habit and a normal evolutionary cycle. The Cariniantum legalis Gallesiosum equilibrium, corresponds however, to a provisory association, because if the moving of soil by torrential rains should cease it would become possible…

Factors influencing the development of Plasmodium gallinaceum in Aedes fluviatilis

Aedes fluviatilis is susceptible to infection by Plasmodium gallinaceum and is a convenient insect host for the malaria parasite in countries where Aedees aegypti cannot be maintained in laboratories. In South America, for instance, the rearing of A. aegypti the main vector of urban yellow fever, is not advaisable because of the potential health hazard it represents. Our results of the comparative studies carried out between the sporogonic cycle produced with two lines of P. gallinaceum parasites into A. fuviatilis were as follows. As proved for A. aegypti, mosquito infection rates were variable when A. fluviatilis blood-fed on chicks infected with and old syringe-passaged strain of P. gallinaceum. Oocysts developed in 41% of those mosquitos and the mean peak of oocyst production was 56 per stomach. Salivary gland infections developed in about 6% of the mosquitos. The course of sporogony was unrelated to the size of the inoculum administered to chicks or to the route by which the birds were infected. The development of infected salivary glands was unrelated to oocyst production. Sporogony of P. gallinaceum was more uniform when mosquitos blood-fed on chicks infected with a sporozoite-passaged strain. Oocysts developed in about 50% of those mosquitoes and the mean peak of oocyst production was 138 per stomach, with some individuals having as many as 600-800 oocysts. Infected salivary glands developed in a mean of 27% of the mosquitos but, in some batches, was a high as 50%. Patterns of salivary gland parasitism were similar to those of oocyst production. The course of sporogony of P. gallinaceum in A. fluviatilis is analized in relation to degree of parasitemia and gametocytemia in the vertebrate host.