Large-scale patterns in morphological diversity and species assemblages in Neotropical Triatominae (Heteroptera: Reduviidae)

We analysed the spatial variation in morphological diversity (MDiv) and species richness (SR) for 91 species of Neotropical Triatominae to determine the ecological relationships between SR and MDiv and to explore the roles that climate, productivity, environmental heterogeneity and the presence of biomes and rivers may play in the structuring of species assemblages. For each 110 km x 110 km-cell on a grid map of America, we determined the number of species (SR) and estimated the mean Gower index (MDiv) based on 12 morphological attributes. We performed bootstrapping analyses of species assemblages to identify whether those assemblages were more similar or dissimilar in their morphology than expected by chance. We applied a multi-model selection procedure and spatial explicit analyses to account for the association of diversity-environment relationships. MDiv and SR both showed a latitudinal gradient, although each peaked at different locations and were thus not strictly spatially congruent. SR decreased with temperature variability and MDiv increased with mean temperature, suggesting a predominant role for ambient energy in determining Triatominae diversity. Species that were more similar than expected by chance co-occurred near the limits of the Triatominae distribution in association with changes in environmental variables. Environmental filtering may underlie the structuring of species assemblages near their distributional limits.

The use of saliva as a practical and feasible alternative to urine in large-scale screening for congenital cytomegalovirus infection increasesinclusion and detection rates

INTRODUCTION: Although urine is considered the gold-standard material for the detection of congenital cytomegalovirus (CMV) infection, it can be difficult to obtain in newborns. The aim of this study was to compare the efficiency of detection of congenital CMV infection in saliva and urine samples. METHODS: One thousand newborns were included in the study. Congenital cytomegalovirus deoxyribonucleic acid (DNA) was detected by polymerase chain reaction (PCR). RESULTS: Saliva samples were obtained from all the newborns, whereas urine collection was successful in only 333 cases. There was no statistically significant difference between the use of saliva alone or saliva and urine collected simultaneously for the detection of CMV infection. CONCLUSIONS: Saliva samples can be used in large-scale neonatal screening for CMV infection.

Assessment of schistosomiasis in the semi-arid Northeast region of Brazil: the São Francisco River large-scale water transposition project

Abstract: INTRODUCTION Risk of schistosomiasis expansion to semi-arid northeastern Brazil under the influence of the Integration Project of the São Francisco River (IPSFR) was assessed. METHODS: Stool examinations of schoolchildren, epidemiological investigation, and survey of the local host snail Biomphalaria straminea were performed in five IPSFR municipalities. RESULTS Six of 4,770 examined schoolchildren were egg-positive for Schistosoma mansoni. Biomphalaria straminea was widespread, but not naturally infected with S. mansoni. Snails experimentally exposed to two laboratory S. mansoni strains yielded infection indices of 1-4.5%. CONCLUSIONS: There is evidence of active schistosomiasis transmission in the area; thus, intensive surveillance actions are required.

PRELIMINARY RESULTS OF A LARGE-SCALE TREE INVENTORY OF UPLAND RAIN FOREST IN THE CENTRAL AMAZON

A large-scale inventory of trees > 10cm DBH was conducted in the upland "terra firme" rain forest of the Distrito Agropecuário da SUFRAMA (Manaus Free Zone Authority Agricultural District) approximately 65Km north of the city of Manaus (AM), Srasil. Thegeneral appearance and structure of the forest is described together with local topography and soil texture. Thepreliminary results of the Inventory provide a minimum estimate of 698 tree species in 53 families in the 40Km radius sampled, including 17 undescribed species. Themost numerically abundant families, Lecythidaceae, Leguminosae, 5apotaceae and Burseraceae as also among the most species rich families. One aspect of this diverse assemblage is the proliferation of species within certain genera, Including 26 genera In 17 families with 6 or more species or morphospecies. Most species have very low abundances of less than 1 tree per hectare. While more abundant species do exist at densities ranging up to a mean of 12 trees per ha, many have clumped distributions leading to great variation in local species abundance. The degree of similarity between hectare samples based int the Coefficient of Community similarity Index varies widely over different sample hectares for five ecologically different families. Soil texture apparently plays a significant role In determining species composition in the different one hectare plots examined while results for other variable were less consistent. Greater differences in similarity indices are found for comparisons with a one hectare sample within the same formation approximately 40Km to the south. It is concluded that homogeneity of tree community composition within this single large and diverse yet continuous upland forest formation can not be assumed.

Malaria diagnosis from pooled blood samples: comparative analysis of real-time PCR, nested PCR and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large-scale

Malaria diagnoses has traditionally been made using thick blood smears, but more sensitive and faster techniques are required to process large numbers of samples in clinical and epidemiological studies and in blood donor screening. Here, we evaluated molecular and serological tools to build a screening platform for pooled samples aimed at reducing both the time and the cost of these diagnoses. Positive and negative samples were analysed in individual and pooled experiments using real-time polymerase chain reaction (PCR), nested PCR and an immunochromatographic test. For the individual tests, 46/49 samples were positive by real-time PCR, 46/49 were positive by nested PCR and 32/46 were positive by immunochromatographic test. For the assays performed using pooled samples, 13/15 samples were positive by real-time PCR and nested PCR and 11/15 were positive by immunochromatographic test. These molecular methods demonstrated sensitivity and specificity for both the individual and pooled samples. Due to the advantages of the real-time PCR, such as the fast processing and the closed system, this method should be indicated as the first choice for use in large-scale diagnosis and the nested PCR should be used for species differentiation. However, additional field isolates should be tested to confirm the results achieved using cultured parasites and the serological test should only be adopted as a complementary method for malaria diagnosis.

Short-term storage in vitro and large-scale propagation of grapevine genotypes

The objective of this work was to evaluate the large-scale propagation of grapevine genotypes after short-term storage in vitro. Microshoots from ten grapevine genotypes were used. The following storage temperatures were evaluated: 10, 20, and 25°C. After short-term storage, the shoots were propagated in up to five successive subcultures, to assess the large-scale propagation of the germplasm maintained under conditions of minimal growth. The propagated shoots were rooted in different concentrations of indolbutiric acid (IBA) and acclimatized in greenhouse. The best temperature for short-term storage in vitro and survival of the genotypes was 20°C. In the propagation phase, the highest number of shoots per explant was found in the subcultures 4 and 5, with averages of 4.9 and 4.8 shoots per explant, respectively. In the rooting phase, the best results for number of roots were obtained using a culture medium supplemented with 0.4 µmol L-1 of IBA, with an average of three roots per shoot. During the acclimation phase, a survival rate higher than 95% was achieved after 30 days in the greenhouse. Grapevine genotypes maintained for six months in vitro, at 20ºC, can be micropropagated in large scale.

Large-scale production and purification of recombinant protein from an insect cell/baculovirus system in Erlenmeyer flasks: application to the chicken poly(ADP-ribose) polymerase catalytic domain

A simple and inexpensive shaker/Erlenmeyer flask system for large-scale cultivation of insect cells is described and compared to a commercial spinner system. On the basis of maximum cell density, average population doubling time and overproduction of recombinant protein, a better result was obtained with a simpler and less expensive bioreactor consisting of Erlenmeyer flasks and an ordinary shaker waterbath. Routinely, about 90 mg of pure poly(ADP-ribose) polymerase catalytic domain was obtained for a total of 3 x 109 infected cells in three liters of culture

Cross-scale determinants of palm species distribution

We investigate palm species distribution, richness and abundance along the Mokoti, a seasonally-dry river of southeastern Amazon and compare it to the patterns observed at a large scale, comprising the entire Brazilian territory. A total of 694 palms belonging to 10 species were sampled at the Mokoti River basin. Although the species showed diverse distribution patterns, we found that local palm abundance, richness and tree basal area were significantly higher from the hills to the bottomlands of the study region, revealing a positive association of these measures with moisture. The analyses at the larger spatial scale also showed a strong influence of vapor pressure (a measure of moisture content of the air, in turn modulated by temperature) and seasonality in temperature: the richest regions were those where temperature and humidity were simultaneously high, and which also presented a lower degree of seasonality in temperature. These results indicate that the distribution of palms seems to be strongly associated with climatic variables, supporting the idea that, by 'putting all the eggs in one basket' (a consequence of survival depending on the preservation of a single irreplaceable bud), palms have become vulnerable to extreme environmental conditions. Hence, their distribution is concentrated in those tropical and sub-tropical regions with constant conditions of (mild to high) temperature and moisture all year round.

Evaluation of temporal, seasonal and geographic stability of the molluscicidal property of Euphorbia splendens latex

Laboratory tests with aqueous solutions of Euphorbia splendens var. hislopii latex have demonstrated seasonal stability of the molluscicidal principle, with LD90 values of 1.14 ppm (spring), 1.02 ppm (fall), 1.09 ppm (winter), and 1.07 ppm (summer) that have been determined against Biomphalaria tenagophila in the field. Assays on latex collected in Belo Horizonte and Recife yielded LD90 values similar to those obtained with the reference substance collected in Rio de Janeiro (Ilha do Governador), demonstrating geographic stability of the molluscicidal effect. The molluscicidal action of aqueous dilutions of the latex in natura, centrifuged (precipitate) and lyophilized, was stable for up to 124 days at room temperature (in natura) and for up to 736 days in a common refrigerator at 10 to 12ºC (lyophilized product). A 5.0 ppm solution is 100% lethal for snails up to 13 days after preparation, the effect being gradually lost to almost total inactivity by the 30th day. This observation indicated that the active principle is instable. These properties together with the wide distribution of the plant, its resistance and adaptation to the tropical climate, its easy cultivation and the easy obtention of latex and preparation of the molluscicidal solution, make this a promising material for large-scale use in the control of schistosomiasis

Scientific authorships and collaboration network analysis on Chagas disease: papers indexed in PubMed (1940-2009)

Chagas disease is a chronic, tropical, parasitic disease, endemic throughout Latin America. The large-scale migration of populations has increased the geographic distribution of the disease and cases have been observed in many other countries around the world. To strengthen the critical mass of knowledge generated in different countries, it is essential to promote cooperative and translational research initiatives. We analyzed authorship of scientific documents on Chagas disease indexed in the Medline database from 1940 to 2009. Bibliometrics was used to analyze the evolution of collaboration patterns. A Social Network Analysis was carried out to identify the main research groups in the area by applying clustering methods. We then analyzed 13,989 papers produced by 21,350 authors. Collaboration among authors dramatically increased over the study period, reaching an average of 6.2 authors per paper in the last five-year period. Applying a threshold of collaboration of five or more papers signed in co-authorship, we identified 148 consolidated research groups made up of 1,750 authors. The Chagas disease network identified constitutes a "small world," characterized by a high degree of clustering and a notably high number of Brazilian researchers.

On the influences of the El Niño, La niña and Atlantic Dipole Paterni on the Amazonian Rainfall during 1960-1998

The influence of the large-scale climatic variability dominant modes in the Pacific and in the Atlantic on Amazonian rainfall is investigated. The composite technique of the Amazon precipitation anomalies is used in this work. The basis years for these composites arc those in the period 1960-1998 with occurrences of extremes in the Southern Oscillation (El Niño or La Niña) and the north/south warm (or cold) sea surface temperature (SST) anomalies dipole pattern in the tropical Atlantic. Warm (cold) dipole means positive (negative) anomalies in the tropical North Atlantic and negative (positive) anomalies in the tropical South Atlantic. Austral summer and autumn composites for extremes in the Southern Oscillation (El Niño or La Niña) and independently for north/south dipole pattern (warm or cold) of the SST anomalies in the tropical Atlantic present values (magnitude and sign) consistent with those found in previous works on the relationship between Amazon rainfall variations and the SST anomalies in the tropical Pacific and Atlantic. However, austral summer and autumn composites for the years with simultaneous occurrences of El Niño and warm north/south dipole of the SST anomalies in the tropical Atlantic show negative precipitation anomalies extending eastward over the center-eastern Amazon. This result indicates the important role played by the tropical Atlantic in the Amazon anomalous rainfall distribution.

Effects of forest structure components on the occurence, group size and density of groups of bare-face tamarin (Saguinus bicolor - primates: Callitrichinae) in Central Amazonia

This study analyzed the influence of forest structural components on the occurence, size and density of groups of Bare-face Tamarin (Saguinus bicolor) - the most threatened species in the Amazon - and produced the first map of distribution of groups in large-scale spatial within the area of continuous forest. Population censuses were conducted between November 2002 and July 2003, covering 6400 hectares in the Ducke Reserve, Manaus-AM, Brazil. Groups of S. bicolor were recorded 41 times accordingly distributed in the environments: plateau (20); slopes (12); and lowlands (09). The mean group size was 4.8 indiv./group, and ranged from 2 to 11 individuals. In the sites where the groups were recorded, and in an equivalent number of sites where no tamarins were found located at least 500 m from those where they had been recorded, we placed 50 m x 50 m plots to record the following forest structural components: abundance of trees; abundance of lianas; abundance of fruiting trees and lianas; abundance of snags; abundance of logs; percentage of canopy opening; leaf litter depth; and altitude. Bare-face Tamarin more often uses areas with lower abundance of forest logs, smaller canopy opening and with higher abundance of snags, areas in the forest with smaller canopy opening present higher density of S. bicolor groups. Apparently this species does not use the forest in a random way, and may select areas for its daily activities depending on the micro-environmental heterogeneity produced by the forest structural components.

A determinação dos números de indivíduos mínimos necessários na experimentação genética

The main object of the present paper consists in giving formulas and methods which enable us to determine the minimum number of repetitions or of individuals necessary to garantee some extent the success of an experiment. The theoretical basis of all processes consists essentially in the following. Knowing the frequency of the desired p and of the non desired ovents q we may calculate the frequency of all possi- ble combinations, to be expected in n repetitions, by expanding the binomium (p-+q)n. Determining which of these combinations we want to avoid we calculate their total frequency, selecting the value of the exponent n of the binomium in such a way that this total frequency is equal or smaller than the accepted limit of precision n/pª{ 1/n1 (q/p)n + 1/(n-1)| (q/p)n-1 + 1/ 2!(n-2)| (q/p)n-2 + 1/3(n-3) (q/p)n-3... < Plim - -(1b) There does not exist an absolute limit of precision since its value depends not only upon psychological factors in our judgement, but is at the same sime a function of the number of repetitions For this reasen y have proposed (1,56) two relative values, one equal to 1-5n as the lowest value of probability and the other equal to 1-10n as the highest value of improbability, leaving between them what may be called the "region of doubt However these formulas cannot be applied in our case since this number n is just the unknown quantity. Thus we have to use, instead of the more exact values of these two formulas, the conventional limits of P.lim equal to 0,05 (Precision 5%), equal to 0,01 (Precision 1%, and to 0,001 (Precision P, 1%). The binominal formula as explained above (cf. formula 1, pg. 85), however is of rather limited applicability owing to the excessive calculus necessary, and we have thus to procure approximations as substitutes. We may use, without loss of precision, the following approximations: a) The normal or Gaussean distribution when the expected frequency p has any value between 0,1 and 0,9, and when n is at least superior to ten. b) The Poisson distribution when the expected frequecy p is smaller than 0,1. Tables V to VII show for some special cases that these approximations are very satisfactory. The praticai solution of the following problems, stated in the introduction can now be given: A) What is the minimum number of repititions necessary in order to avoid that any one of a treatments, varieties etc. may be accidentally always the best, on the best and second best, or the first, second, and third best or finally one of the n beat treatments, varieties etc. Using the first term of the binomium, we have the following equation for n: n = log Riim / log (m:) = log Riim / log.m - log a --------------(5) B) What is the minimun number of individuals necessary in 01der that a ceratin type, expected with the frequency p, may appaer at least in one, two, three or a=m+1 individuals. 1) For p between 0,1 and 0,9 and using the Gaussean approximation we have: on - ó. p (1-p) n - a -1.m b= δ. 1-p /p e c = m/p } -------------------(7) n = b + b² + 4 c/ 2 n´ = 1/p n cor = n + n' ---------- (8) We have to use the correction n' when p has a value between 0,25 and 0,75. The greek letters delta represents in the present esse the unilateral limits of the Gaussean distribution for the three conventional limits of precision : 1,64; 2,33; and 3,09 respectively. h we are only interested in having at least one individual, and m becomes equal to zero, the formula reduces to : c= m/p o para a = 1 a = { b + b²}² = b² = δ2 1- p /p }-----------------(9) n = 1/p n (cor) = n + n´ 2) If p is smaller than 0,1 we may use table 1 in order to find the mean m of a Poisson distribution and determine. n = m: p C) Which is the minimun number of individuals necessary for distinguishing two frequencies p1 and p2? 1) When pl and p2 are values between 0,1 and 0,9 we have: n = { δ p1 ( 1-pi) + p2) / p2 (1 - p2) n= 1/p1-p2 }------------ (13) n (cor) We have again to use the unilateral limits of the Gaussean distribution. The correction n' should be used if at least one of the valors pl or p2 has a value between 0,25 and 0,75. A more complicated formula may be used in cases where whe want to increase the precision : n (p1 - p2) δ { p1 (1- p2 ) / n= m δ = δ p1 ( 1 - p1) + p2 ( 1 - p2) c= m / p1 - p2 n = { b2 + 4 4 c }2 }--------- (14) n = 1/ p1 - p2 2) When both pl and p2 are smaller than 0,1 we determine the quocient (pl-r-p2) and procure the corresponding number m2 of a Poisson distribution in table 2. The value n is found by the equation : n = mg /p2 ------------- (15) D) What is the minimun number necessary for distinguishing three or more frequencies, p2 p1 p3. If the frequecies pl p2 p3 are values between 0,1 e 0,9 we have to solve the individual equations and sue the higest value of n thus determined : n 1.2 = {δ p1 (1 - p1) / p1 - p2 }² = Fiim n 1.2 = { δ p1 ( 1 - p1) + p1 ( 1 - p1) }² } -- (16) Delta represents now the bilateral limits of the : Gaussean distrioution : 1,96-2,58-3,29. 2) No table was prepared for the relatively rare cases of a comparison of threes or more frequencies below 0,1 and in such cases extremely high numbers would be required. E) A process is given which serves to solve two problemr of informatory nature : a) if a special type appears in n individuals with a frequency p(obs), what may be the corresponding ideal value of p(esp), or; b) if we study samples of n in diviuals and expect a certain type with a frequency p(esp) what may be the extreme limits of p(obs) in individual farmlies ? I.) If we are dealing with values between 0,1 and 0,9 we may use table 3. To solve the first question we select the respective horizontal line for p(obs) and determine which column corresponds to our value of n and find the respective value of p(esp) by interpolating between columns. In order to solve the second problem we start with the respective column for p(esp) and find the horizontal line for the given value of n either diretly or by approximation and by interpolation. 2) For frequencies smaller than 0,1 we have to use table 4 and transform the fractions p(esp) and p(obs) in numbers of Poisson series by multiplication with n. Tn order to solve the first broblem, we verify in which line the lower Poisson limit is equal to m(obs) and transform the corresponding value of m into frequecy p(esp) by dividing through n. The observed frequency may thus be a chance deviate of any value between 0,0... and the values given by dividing the value of m in the table by n. In the second case we transform first the expectation p(esp) into a value of m and procure in the horizontal line, corresponding to m(esp) the extreme values om m which than must be transformed, by dividing through n into values of p(obs). F) Partial and progressive tests may be recomended in all cases where there is lack of material or where the loss of time is less importent than the cost of large scale experiments since in many cases the minimun number necessary to garantee the results within the limits of precision is rather large. One should not forget that the minimun number really represents at the same time a maximun number, necessary only if one takes into consideration essentially the disfavorable variations, but smaller numbers may frequently already satisfactory results. For instance, by definition, we know that a frequecy of p means that we expect one individual in every total o(f1-p). If there were no chance variations, this number (1- p) will be suficient. and if there were favorable variations a smaller number still may yield one individual of the desired type. r.nus trusting to luck, one may start the experiment with numbers, smaller than the minimun calculated according to the formulas given above, and increase the total untill the desired result is obtained and this may well b ebefore the "minimum number" is reached. Some concrete examples of this partial or progressive procedure are given from our genetical experiments with maize.

Dried blood spots collected on filter paper: an international resource for the diagnosis and genetic characterization of human immunodeficiency virus Type-1

The collection of dried blood spots (DBS) on filter paper provides a powerful approach for the development of large-scale, population-based screening programs. DBS methods are particularly valuable in developing countries and isolated rural regions where resources are limited. Large numbers of field specimens can be economically collected and shipped to centralized reference laboratories for genetic and (or) serological analysis. Alternatively, the dried blood can be stored and used as an archival resource to rapidly establish the frequency and distribution of newly recognized mutations, confirm patient identity or track the origins and emergence of newly identified pathogens. In this report, we describe how PCR-based technologies are beginning to interface with international screening programmes for the diagnosis and genetic characterization of human immunodeficiency virus type 1 (HIV-1). In particular, we review recent progress using DBS specimens to resolve the HIV-1 infection status of neonates, monitor the genetic evolution of HIV-1 during early infancy and establish a sentinel surveillance system for the systematic monitoring of HIV-1 genetic variation in Asia.

Trypanorhynch Cestodes of Commercial Fishes from Northeast Brazilian Coastal Waters

A large scale investigation on trypanorhynch cestode infestation of tropical marine fishes was carried out along the Northeast Brazilian coast in the summer of 1991 and 1993. A total of 798 fish specimens belonging to 57 species and 30 families were examined. Metacestodes of 11 different trypanorhynchs were found: Callitetrarhynchus gracilis, Dasyrhynchus giganteus, Grillotia sp., Nybelinia edwinlintoni, N. indica, N. senegalensis, Nybelinia c.f. lingualis, Otobothrium cysticum, Pseudolacistorhynchus noodti, Pseudotobothrium dipsacum and Pterobothrium kingstoni. Scanning electron microscopy was used to clarify details of the tentacular armature of some species. Rose-thorn shaped hooklets, regularly arranged like microtriches, are described from the bothridial surface of N. edwinlintoni. Of the 57 fish species, 15 harboured trypanorhynch cestodes. Of these the mullid Pseudupeneus maculatus was the most heavily infested fish species, harbouring 5 different trypanorhynch species. P. noodti in P. maculatus had the highest prevalence (87%) and intensity (maximum = 63) of infestation. C. gracilis was the parasite with the lowest host-specificity. It could be isolated from 10 fish species. The cestode fauna of the Northeast Brazilian coast appears to be similar to that of the West African coast. Five of the trypanorhynch cestodes found during this study are common to both localities. The two single cases of intra musculature infestation in Citharichthys spilopterus and Haemulon aurolineatum by trypanorhynch cestodes indicate that marketability of the investigated commercially exploited fish species is inconsequential.