Mendelian inheritance, genetic linkage, and genotypic disequilibrium for nine microsatellite loci in Cariniana estrellensis (Raddi) Kuntze (Lecythidaceae).

Cariniana estrellensis is one of the largest trees found in Brazilian tropical forests. The species is typical of advanced stages of succession, characteristic of climax forests, and essential in genetic conservation and environmental restoration plans. In this study, we assessed Mendelian inheritance, genetic linkage, and genotypic disequilibrium in nine microsatellite loci for a C. estrellensis population. We sampled and genotyped 285 adult trees and collected seeds from 20 trees in a fragmented forest landscape in Brazil. Based on maternal genotypes and their seeds, we found no deviation from the expected 1:1 Mendelian segregation and no genetic linkage between pairwise loci. However, for adults, genotypic disequilibrium was detected for four pairs of loci, suggesting that this result was not caused by genetic linkage. Based on these results, we analyzed microsatellite loci that are suitable for use in population genetic studies assessing genetic diversity, mating system, and gene flow in C. estrellensis populations.

Short Communication Mendelian inheritance, linkage, and genotypic disequilibrium in microsatellite loci of Hymenaea stigonocarpa Mart. ex Hayne (Fabaceae-Caesalpinioideae).

Hymenaea stigonocarpa is a deciduous and monoecious Neotropical tree species pollinated by bats. Due to overexploitation and habitat destruction, the population size has drastically diminished in nature. No previous study has investigated Mendelian inheritance, linkage, and genotypic disequilibrium in the available microsatellite markers in this species. So, our aim was to estimate these parameters using six microsatellite loci in a sample of 470 adults and 219 juveniles from two populations of H. stigonocarpa. In addition, 30 seeds per tree from 35 seed-trees were collected. Each seed was kept record of the seed-trees and fruit origin. Based on the six microsatellite loci, we found that only 10.6% of the cases showed significant deviations from Mendelian segregation and 15.3% showed linkage. We detected no evidence of genotypic disequilibrium between the loci in the adult trees or juveniles. Thus, our results suggest that these loci can be used with great accuracy in future genetic analyses of H. stigonocarpa populations.

Nitrogen and phosphorus flux from the production of Nile tilapia through the application of environmental indicators.

We aimed in this study utilize environmental indicators as a quantitative method to evaluate and discuss the nitrogen (TN) and phosphorus (TP) flux by a production stage grow-out (termination) of Nile tilapia (Oreochromis niloticus) in fishpond. The TN and TP load, the mass balance, the input of TN and TP via feed and the converted nutrients in fish biomass are the environmental indicators applied in this study. During the production cycle (128 days), the system exported 15,931 g TN and 4,189 g TP that were related to the amount of feed supplied (r Pearson = 0.8825 and r = 0.8523, respectively), corroborated by the feed conversion ratio (1.61:1). The indicators showed that 26% TN and 45% TP were reversed into fish biomass, 62% TN and 40% TP were retained in the fishpond, and 12% TN and 15% TP were exported via effluent. The largest contribution of nutrients generated by the system and exported via effluent was observed in phase III and IV. This result is supported by the feed conversion ratio 2.14 and 2.21:1 obtained at this phase, a fact explained by the amount of feed offered and the fish metabolism. Application of environmental indicators showed to be an efficient tool to quantify flux of TN and TP produced during the grow-out period of Nile tilapia and therefore, guide management practices more sustainable. Concerning the environmental sustainability of the activity the implementation of best management practices such as the better control of the feed amount offered would lead to a smaller loss of TN and TP to the water. Furthermore, the use of better quality feeds would allow greater nutrient assimilation efficiency.

Environmental indicators in effluent assessment of rainbow trout (Oncorhynchus mykiss) reared in raceway system through phosphorus and nitrogen.

The phosphorus and nitrogen discharge via effluent of intensive trout farming system was quantified through the use of environmental indicators. The nutrient loads, the mass balance, the estimated amount of nutrients in feed and the amount of nutrients converted in fish biomass were calculated based on the concentrations of phosphorus (P) and nitrogen (N) in the feed and in the water. Of the offered feed, 24.75 kg were available as P and 99.00 kg as N, of these, 9.32 kg P (38%) and 29.12 kg N (25%) were converted into fish biomass and 15.43 kg P (62%) and 69.88 kg N (75%) were exported via effluent. The loads and the mass balance show the excessive discharge of nutrients via effluent, corroborated by the feed conversion ratio (2.12:1) due to the low efficiency of feed utilization, therefore, it is proposed the use of this zootechnical parameter as environmental indicator. In addition, feed management practices are not adequate, highlighting the low frequency of feeding during the day, excessive amount and low quality of feed offered. These results demonstrate the need for adequate feed management and the need for careful monitoring of effluent.

HMGB1 and inflammatory cytokines in experimental acute lung injury induced in rabbits / [HMGB1 e citocinas inflamatórias em lesão pulmonar aguda experimental induzida em coelhos]

The aim of this work was to measure HMGB1, TNF-alpha, and IL-8 in bronchoalveolar lavage (BAL), serum and TLR2 and TLR4mRNA expression in lung tissue of rabbits with two grades of acute lung injury (ALI). The animals were randomly assigned to groups with severe (S) and mild/moderate (MM) ALI, induced with warm saline, and a control group. HMGB1, TNF-alpha, IL-8, TLR2mRNA and TLR4mRNA were measured after ALI induction. The results showed increased levels of IL-8, TNF-alpha, HMGB1 and TLR4mRNA in the ALI groups. HMGB1, IL-8 and TNF-alpha concentrations in BAL were higher in S compared MM. Increased TLR4mRNA was observed in S and MM versus control. The results suggest an early participation of HMGB1 in ALI together with IL-8 and TNF-alpha and association with severity. TLR4 has early expression and role in ALI pathophysiology but is not associated with severity.(AU)

O objetivo deste trabalho é determinar os níveis de HMGB1, TNF-alfa e IL-8 no lavado broncoalveolar (BAL), bem como quantificar a expressão sérica de TLR2 e TLR4 mRNA em tecido pulmonar de coelhos com dois graus de lesão pulmonar aguda (LPA). Os animais foram distribuídos aleatoriamente em grupos com LPA grave (S) e leve / moderada (MM), induzidas com solução salina morna, e um grupo controle. HMGB1, TNF-alfa, IL-8, TLR2mRNA e TLR4mRNA foram medidos após a indução de LPA e quatro horas de ventilação mecânica. Os resultados mostraram níveis aumentados de IL-8, TNF-alfa, HMGB1 e TLR4mRNA nos grupos com LPA. As concentrações de HMGB1, IL-8 e TNF-alfa no LBA foram maiores no S comparado ao MM. Aumento de TLR4mRNA foi observado em S e MM versus controle. Os resultados sugerem uma participação precoce da HMGB1 na LPA em conjunto com IL-8 e TNF-alfa e associação com a gravidade da LPA. O TLR4 foi expresso na ALI e possivelmente possui papel precoce na fisiopatologia da LPA, mas sem associação com a gravidade.(AU)

An epidemic of yellow fever in Central Brazil, 1972-1973. II. Ecological studies.

Results of ecological studies undertaken during the 1972-1973 epidemic of yellow fever (YF) in Goiás State, Brazil, suggest that mosquitoes of the genus Haemagogus were the only vectors infected with YF virus. Nine isolations of the agent were made from 1,688 Haemagogus spp. caught within or near forests from 27 January to 3 March 1973. Seven of these isolations cane from mosquitoes collected on the forest floor. No YF virus isolation was made from 791 Haemagogus leucocelaenus or 1,096 Sabethes chloropterus. Haemagogus were caught both in and outside houses located 500 m from the forest, although at a lower rate than in the forest. Haemagogus collection rates in a secondary growth forest were similar to or even higher than those in primary forests. YF virus was recovered from a Cebus monkey and hemagglutination-inhibition and/or neutralizing antibody to YF virus was found in seven (30.4%) of 23 primates examined. There was, however, little evidence of sick or dying monkeys. Among other wild-caught animals only a single marsupial (Caluromys) had antibodies to the virus. These data show that sylvan YF is not confined to the forest, nor always associated with conspicuous epizootics.

An epidemic of yellow fever in central Brazil. 1972-1973. I. Epidemiological studies.

An epidemic of jungle yellow fever occurred in Goiás State, Brazil, between December 1972 and March 1973. Laboratory confirmed cases were observed in 36 counties located in the central and southern parts of the State. Seventy-one cases were proved, of which 44 were fatal. The diagnosis was made on the basis of pathology, serology, and virus isolation. Besides yellow fever, malaria and viral hepatitis were present, and in two fatal cases there was malarial pigment in the liver in addition to the specific lesions associated with yellow fever virus infection. The fact that male patients strikingly outnumbered females (9:1) and that young adults were predominantly affected indicates that transmission occurred mainly inside or adjacent to the forests. The lack of cases in urban areas can be attributed to the absence of Aedes aegypti in these areas. Yellow fever complement-fixing antibody in high titers was found in 18 of 1,201 (1.4%) persons living in eight counties of the affected area. This finding suggests that at least 21,000 persons out of the 1.5 million rural inhabitants of the three districts where the epidemic occurred had been infected by the virus. The epidemic subsided following an intensive vaccination campaign, and the last four cases were observed in March 1973.

An epidemic of sylvatic yellow fever in the southeast region of Maranhao State, Brazil, 1993-1994: epidemiologic and entomologic findings.

Yellow fever virus transmission was very active in Maranhao State in Brazil in 1993 and 1994. An investigation was carried out to evaluate the magnitude of the epidemic. In 1993, a total of 932 people was examined for yellow fever from Maranhao: 70 were positive serologically, histopathologically, and/or by virus isolation, and another four cases were diagnosed clinically and epidemiologically. In Mirador (17,565 inhabitants), the incidence was 3.5 per 1,000 people (case fatality rate [number of deaths/number of cases diagnosed] = 16.4%), while in a rural yellow fever risk area (14,659 inhabitants), the incidence was 4.2 and the case-fatality rate was 16.1% (10 of 62). A total of 45.2% (28 of 62) asymptomatic infections were registered. In 1994, 49 serum samples were obtained and 16 cases were confirmed (two by virus isolation, two by seroconversion, and 12 by serology). No fatal cases were reported. In 1993, 936 potential yellow fever vectors were captured in Mirador and a single strain was isolated from a pool of Haemagogus janthinomys (infection rate = 0.16%). In 1994, 16 strains were isolated from 1,318 Hg. janthinomys (infection rate = 1.34%) and one Sabethes chloropterus (infection rate = 1.67%). Our results suggest that this was the most extensive outbreak of yellow fever in the last 20 years in Brazil. It is also clear that the lack of vaccination was the principal reason for the epidemic, which occurred between April and June, during the rainy season, a period in which the mosquito population in the forest increases.

Simulium species of the amazonicum group as vectors of Mansonella ozzardi in the Brazilian Amazon.

Experimental infection with Mansonella ozzardi of common haematophagous Diptera collected at a Ticuna Indian village on the upper reaches of the Solimões river in the Brazilian Amazon, showed that Simulium amazonicum and Simulium n.sp. are capable of supporting full development of the parasite. Natural infections with this filaria were found in both species including infective larvae in Simulium n.sp. No development of M. ozzardi occurred in Mansonia amazonensis, Culicoides insinuatus or Lepiselaga crassipes (Tabanidae). The dimensions of developing larvae of M. ozzardi in both species of black-fly were recorded. Infective larvae of this species may easily be distinguished from those of Onchocerca volvulus, also transmitted in the Amazon by a species closely resembling S. amazonicum, by the presence of a bifid tail and higher anal ratio in M. ozzardi.

The concentration of Mansonella ozzardi microfilariae in skin capillaries.

The densities of microfilariae of Mansonella ozzardi in different regions of the body of infected individuals were studied. The density in venous blood closely approximated to that found in finger capillaries, but was significantly lower than the densities in capillaries of the scapular region and buttocks. A possible explanation for this concentration of microfilariae is the lack of glomi in these two sites, which consequently have a lower flow of blood than areas where glomi occur and where microfilarial densities are lower. No relationship was found between blood and skin densities in the buttocks and scapular region, probably due to the inadequacy of the skin snip method used for the detection of this species of filaria. It was concluded that the most acceptable detection method for M. ozzardi remains the sampling of finger capillary blood. It could not be decided, due to the paucity of data, whether any relationship exists between skin microfilarial densities and the biting sites of the vectors.

Experimental Trypanosoma evansi infection in South American coati (Nasua nasua): hematological, biochemical and histopathological changes.

The course of an experimental Trypanosoma evansi infection in coatis (Nasua nasua, carnivora, Procyonidae) was followed for 262 days. Hematological analysis of the infected coatis revealed a marked decline in hemoglobin, packed-cell volume, and total erythrocyte count. An intense anemia followed the first wave of parasitemia and persisted until the end of the experimental period. Biochemical analysis showed increased serum levels of alanine aminotransferase and aspartate aminotransferase and decreased albumin. The main histopathological features consisted of myocarditis with the presence of degenerate cardiac fibers and meningoencephalitis. This study has shown that coatis infected with T. evansi develop a chronic disease.

Trypanosoma evansi experimental infection in the South American coati (Nasua nasua): clinical, parasitological and humoral immune response.

The course of Trypanosoma evansi infection in coatis (Carnivora, Procionidae) was followed for 262 days. Parasites were detected in all infected animals from day 2 post infection until the end of the study. No correlation between temperature and parasitemia was observed. Animals of the infected group demonstrated depression, weakness, lethargy and pale mucous membranes. Indirect fluorescent antibody tests detected anti-T. evansi antibodies within 7 to 14 days post infection and showed high levels until the end of the experimental period. The persistent parasitemia in coati and their relative tolerance to clinical signs suggested that this species develops a chronic disease and plays an important role in the epidemiology of trypanosomosis due to T. evansi in enzootic regions.

Dissemination of Leishmania (Viannia) braziliensis.

Destructive human mucocutaneous leishmaniasis may appear many years after the primary cutaneous infection with Leishmania (Viannia) braziliensis. Hamsters (Mesocricetus auratus) were infected with metacyclic L. braziliensis promastigotes. It was found that secondary metastatic visceral lesions could arise from a primary cutaneous lesion, or secondary cutaneous lesions from a primary visceral lesion. Parasites in the viscera were shown to be viable, multiplying and capable of metastasis to either secondary visceral or cutaneous sites. The finding of an early metastasis in the wall of a small cutaneous vessel indicates that dissemination can occur by the haematogenous route. Slow growing organisms in viscera may thus be a source for late metastasis to mucocutaneous sites or for systemic relapse after immunosuppression.

[Histopathology of the localized form of cutaneous leishmaniasis caused by Leishmania (Leishmania) amazonensis]. / Histopatologia da forma localizada de leishmaniose cutânea por Leishmania (Leishmania) amazonensis.

The microscopic changes found in the localized form of the human cutaneous leishmaniasis due to Leishmania (Leishmania) amazonensis are reported. In this form, less known than the diffuse one caused by the same species, the clinical manifestations are identical to those produced by other Leishmania species of the subgenus Viannia. There is, however, in the localized infection by L. (L.) amazonensis, a peculiar feature, only recently discovered: about 50% of the affected individuals are Montenegro-negatives. The main histologic change observed in the skin sections was the presence of groups of macrophages with a large vacuole in the cytoplasm, containing many amastigotes. The microscopic picture is similar to that found in the diffuse form of the disease, the difference being only quantitative. When in large numbers, the macrophages suffers necrosis, which generally starts at the center of the groups. First, in this process, the membrane of the parasitized cells ruptures, and the amastigotes become free; later, both cells and parasites are destroyed. The picture can be seen either in Montenegro-negative or in Montenegro-positive patients. The macrophages with amastigotes may persist in tissues for as long as 6-7 months, while in the infections due to L. (V.) braziliensis the parasites usually disappear in a few weeks.

[Histoplasmoma with pleural effusion. Report of a case]. / Histoplasmoma com derrame pleural. Relato de um caso.

Pleural effusions are rare events accompanying histoplasmosis, and in general they are associated with acute forms of the disease. It is described here a case of histoplasmoma associated with pleuritic pain and pleural effusion.

[Experimental cutaneous leishmaniasis. III. Histopathological aspects of the developmental behavior of the cutaneous lesion induced in Cebus apella (Primates: Cebidae) by Leishmania (Viannia) lainsoni, L. (V.) braziliensis and L. (Leishmania) amazonensis]. / Leishmaniose cutânea experimental. III--Aspectos histopatológicos do comportamento evolutivo da lesão cutânea produzida em Cebus apella (Primates: Cebidae) por Leishmania (Viannia) lainsoni, L. (V.) braziliensis E L. (Leishmania) amazonensis.

We have studied the histopathological aspects related to the evolution of cutaneous lesions experimentally produced in the monkey Cebus apella (Primates: Cebidae) by Leishmania (V.) lainsoni, L. (V.) braziliensis and L. (L.) amazonensis. Microscopical examination of a series of biopsies obtained from these animals showed the kinetics of the cutaneous lesions regarding three species of Leishmania inoculated, as follows: 1) an initial non-specific chronic inflammatory infiltrate; 2) macrophagic nodules; 3) necrosis of parasitized phagocytic cells; 4) epitheliode granuloma; 5) absorption of the necrotic area (sometimes forming "foreign-body granuloma"); 6) a non-specific residual inflammatory infiltration; and 7) cicatrization. These pathological processes are, of course, responsible for both development and resolution of the leishmaniotic lesion. We also discuss some immunopathological mechanisms probably related with the sequential events, and that could be also responsible for the different clinical aspects found in man.

[A fatal case of human pulmonary adiaspiromycosis]. / Caso fatal de adiaspiromicose pulmonar humana.

A fatal case of human pulmonary adiaspiromycosis is reported. The patient, a male farm laborer from Pernambuco, in the Northeastern part of Brazil, had moved to Planaltina, DF., one year prior the onset of his illness. The main clinical manifestations consisted in fever, myalgia, cough and dyspnea. After being sick for five weeks, the patient developed severe respiratory failure and died. Grossly, disseminated nodular lesions, a few millimeters in diameter, were observed throughout the lobes of both lungs. Microscopically, round structures, up to 600 microns in diameter, with a thick membrane, identified as adiaconidia of Chrysosporium parvum var. crescens, were seen in the center of the nodular lesions. These adiaconidia appeared predominantly inside microabscesses or necrotic areas, both surrounded by a granulomatous reaction. The pulmonary alveoli not affected by the nodular lesions contained an inflammatory exudate composed of macrophages and neutrophils. The finding of several non-fatal cases of the disease in the area of the Federal District suggests that adiaspiromycosis is endemic in the Central Brazilian Plateau region, where the dry, warm and windy climate, particularly from August to October, may provide the appropriate environmental conditions for the dissemination of C. parvum conidia.

[Nasofacial zygomycosis. Report of a case of Para state, Brazil]. / Zigomicose nasofacial. Relato de um caso do estado do Pará, Brasil.

A case of nasofacial zygomycosis in a 64-year-old female patient from Barcarena, State of Pará, is reported. This is the second time that the disease is recognized in Northern Brazil. The diagnosis was initially suspected by the direct observation of broad coenocytic hyphae in biopsy specimens of the nasal cavity mucosa. Later, the cause of the disease was confirmed by the mycologial and histopathological findings. Rapid response to the therapy with potassium iodide was achieved, without however a complete resolution of the lesions. The patient still remains under treatment, with itraconazole and iodide.

[Human lagochilascariasis. 3 cases encountered in the Federal District, Brazil]. / Lagoquilascaríase humana. Sobre três casos encontrados no Distrito Federal, Brasil.

Three new cases of human infection with Lagochilascaris minor are reported. All the patients were from the state of Pará (Brazil), living in rural areas or close to the forest. They were admitted to hospitals in the Federal District due to the presence of abscesses in the region of the neck, ear, mastoid process and, in two of them, to the involvement also of the central nervous system. Microscopic examination of tissue samples taken from the lesions showed sections of eggs and worms--in different evolutive stages--identified as Lagochilascaris minor. Larvae and adult worms obtained from existing fistulae proved also to be of the same species. The three patients were treated with anthelmintic drugs and surgical excision of the lesions, with good clinical results. In two of them, however, relapsing occurred, suggesting that the drugs do not destroy the worm eggs, in spite of the apparent healing of the lesions.

[Lymphadenopathies in American cutaneous leishmaniasis: comments on 2 cases]. / Linfadenopatias na leishmaniose tegumentar americana: considerações sobre dois casos.

Two patients, with cervical lymphadenopathies and a presumptive diagnosis of lymphoma or scrofula, were submitted to biopsies of the affected lymph nodes. Unexpectedly, the histological picture revealed a necrotic-granulomatous reaction and the presence of leishmania (amastigotes) in some vacuolated macrophages. The patients were from different endemic areas of leishmaniasis in Brazil, and had no perceptible cutaneous or mucosal lesions. Later, however, one of them developed such lesions, probably as an effect of a treatment for toxoplasmosis. These findings support the idea that the agents of the disease, once in the host organism, would invade the organs of the phagocytic-mononuclear system, there remaining for a long time, maybe for the rest of the host life. Eventually, under the action of several factors, that could modify the host resistance, the parasites migrate to the skin or the mucosal membranes, causing secondary or reactivating lesions.