Isolation of fungi in Musca domestica Linnaeus, 1758 (Diptera: Muscidae) captured at two natural breeding grounds in the municipality of Seropédica, Rio de Janeiro, Brazil

The objective of this study was to isolate and identify fungal species found in natural association with adults of Musca domestica. The adult insects were collected from two natural breeding grounds: hog pens and an urban sanitary landfill. The isolated fungi were identified as: Aspergillus flavus (23.8%), A. niger var. niger (14.4%), Penicillium corylophilum (21.4%), P. fellutanum (11.9%), Cladosporium cladosporoides (4.7%), Fusarium sp. (4.7%), Alternaria alternata (11.9%), Curvularia brachyspora (2.4%), Mycelia sterilia (2.4%) and the Mucorales order (2.4%).

Value of morphotyping for the characterization of Candida albicans clinical isolates

Until recently, morphotyping, a method evaluating fringe and surface characteristics of streak colonies grown on malt agar, has been recommended as a simple and unexpensive typing method for Candida albicans isolates. The discriminatory power and reproducibility of Hunter's modified scheme of Phongpaichit's morphotyping has been evaluated on 28 C. albicans isolates recovered from the oral cavity of asymptomatic human immunodeficiency virus-positive subjects, and compared to two molecular typing methods: randomly amplified polymorphic DNA (RAPD) fingerprinting, and contour clamped homogeneous electric field (CHEF) electrophoretic karyotyping. Morphological features of streak colonies allowed to distinguish 11 different morphotypes while RAPD fingerprinting yielded 25 different patterns and CHEF electrophoresis recognized 9 karyotypes. The discriminatory power calculated with the formula of Hunter and Gaston was 0.780 for morphotyping, 0.984 for RAPD fingerprinting, and 0.630 for karyotyping. Reproducibility was tested using 43 serial isolates from 15 subjects (2 to 6 isolates per subject) and by repeating the test after one year storage of the isolates. While genetic methods generally recognized a single type for all serial isolates from each of the subjects studied, morphotyping detected strain variations in five subjects in the absence of genetic confirmation. Poor reproducibility was demonstrated repeating morphotyping after one year storage of the isolates since differences in at least one character were detected in 92.9% of the strains.

Maintenance and breeding of Thrichomys (Trouessart, 1880) (Rodentia: Echimyidae) in captivity

South American histricognath rodents Thrichomys apereoides laurentius and Thrichomys pachyurus are natural hosts of Trypanosoma cruzi, agent of Chagas disease. We established breeding colonies of these species to serve as experimental models in various parasitological studies. Both species of Thrichomys have all the requirements necessary to become excellent laboratory models: they can be easily maintained in the standard laboratory conditions and breed throughout the year and they do not have any special dietary demands and can be fed by standard food pellets designed for laboratory mice. Both species produce precocious offspring that have their eyes and ears open, teeth erupted, fur well developed, and can eat solid food in the first week of life. T. a. laurentius has larger litter sizes and lower body masses at birth and weaning than T. pachyurus. Moreover, females of T. a. laurentius reach puberty earlier and with lower body mass than T. pachyurus.

Effect of food on immature development, consumption rate, and relative growth rate of Toxorhynchites splendens (Diptera: Culicidae), a predator of container breeding mosquitoes

Food utilization by the larvae of Toxorhynchites splendens (Wiedemann) was studied in the laboratory by offering larvae of Aedes aegypti Linnaeus, Anopheles stephensi (Liston), and Culex quinquefasciatus (Say). Quantitative analyses of data indicated that immature development was significantly faster with increase in food availability. The regression analysis showed that the degrees of the relationship between immature duration (Id) and food availability were higher when offered early instars of prey (first and second instars) than late instars. Consumption rate (Cr) of the predator increased with increase in food availability and this relationship was highly significant when larvae of An. stephensi were offered as food. Consumption rate to food level decreased with increase in the age class of the prey. There was a significant negative correlation between Id and Cr. This aspect helps to increase population turnover of T. splendens in a shorter period when the prey is abundant. Conversely, the predator compensated the loss in daily food intake at low food level by extending Id thereby attains the minimum threshold pupal weight for adult emergence. There was an increase in the relative growth rate (RGR) of the predator when An. stephensi was offered as prey and this was related to the high protein content of the prey per body weight. There was a positive correlation between Cr and RGR. This adaptive life characteristic strategy of this predator is useful for mass-rearing for large scale field release programmes in the control of container breeding mosquitoes is discussed.

Beyond sepsis pathophysiology with cytokines: what is their value as biomarkers for disease severity?

Sepsis is a major challenge in medicine. It is a common and frequently fatal infectious condition. The incidence continues to increase, with unacceptably high mortality rates, despite the use of specific antibiotics, aggressive operative intervention, nutritional support, and anti-inflammatory therapies. Typically, septic patients exhibit a high degree of heterogeneity due to variables such as age, weight, gender, the presence of secondary disease, the state of the immune system, and the severity of the infection. We are at urgent need for biomarkers and reliable measurements that can be applied to risk stratification of septic patients and that would easily identify those patients at the highest risk of a poor outcome. Such markers would be of fundamental importance to decision making for early intervention therapy or for the design of septic clinical trials. In the present work, we will review current biomarkers for sepsis severity and especially the use of cytokines as biomarkers with important pathophysiological role.

Description of Culicoides (Mataemyia) felippebauerae sp. n., Forcipomyia musae immatures, and occurrence of F. genualis, breeding in banana stems in Brazilian Amazonia (Diptera: Ceratopogonidae)

The following three species of Ceratopogonidae were collected breeding in the rhizomatous herb Phenakospermum guyannense Endl., 1833 in the vicinity of Manaus, Brazil, a new species, Culicoides (Mataemyia) felippebauerae Spinelli, Forcipomyia (Forcipomyia) genualis (Loew), and F. (Phytohelea) musae Clastrier & Dellécole. C. (M.) felippebauerae is described and illustrated as adult, pupa, and fourth instar larva, the adult compared with the adult of C. barthi Taveres and Souza and larva and pupa with those of C. dicrourus Wirth & Blanton and C. macieli Tavares & Ruiz, the only species with known immatures in the subgenus. The pupa and fourth instar larva of F. (P.) musae are described and illustrated and compared with immatures of F. (P.) edwardsi Saunders.

Usefulness of PCR-based assays to assess drug efficacy in Chagas disease chemotherapy: value and limitations

One major goal of research on Chagas disease is the development of effective chemotherapy to eliminate the infection from individuals who have not yet developed cardiac and/or digestive disease manifestations. Cure evaluation is the more complex aspect of its treatment, often leading to diverse and controversial results. The absence of reliable methods or a diagnostic gold standard to assess etiologic treatment efficacy still constitutes a major challenge. In an effort to develop more sensitive tools, polymerase chain reaction (PCR)-based assays were introduced to detect low amounts of Trypanosoma cruzi DNA in blood samples from chagasic patients, thus improving the diagnosis and follow-up evaluation after chemotherapy. In this article, I review the main problems concerning drug efficacy and criteria used for cure estimation in treated chagasic patients, and the work conducted by different groups on developing PCR methodologies to monitor treatment outcome of congenital infections as well as recent and late chronic T. cruzi infections.

Breeding sites of Culicoides pachymerus Lutz in the Magdalena River basin, Colombia

The breeding sites of Culicoides pachymerus are described for the first time in western Boyacá Province, Colombia, where this species is a public health problem. In addition to being a nuisance due to its enormous density and its high biting rates, C. pachymerus cause dermatological problems in the human population. Analysis of microhabitats by the sugar flotation technique and the use of emergence traps allowed us to recover 155 larvae of Culicoides spp and 65 adults of C. pachymerus from peridomiciliary muddy substrates formed by springs of water and constant rainwater accumulation. These important findings could aid in the design of integrated control meas-ures against this pest.

Tire-breeding mosquitoes of public health importance along an urbanisation gradient in Buenos Aires, Argentina

Used vehicle tires are a source of mosquito vectors and a means of their introduction and expansion. With the aim of assessing the effects of urbanisation on the main mosquito vectors in temperate Argentina, the infestation levels of Aedes aegypti (L.) and Culex pipiens L. were studied in used tires from highly urbanised cities to low-urbanised small towns in Buenos Aires. Immatures of both species accounted for 96% of the 9,722 individuals collected; the total individuals collected represented seven species. The percentage of water-filled tires containing mosquitoes [container index (CI)] was 33% and the percentage of infested sites [site index (SI)] was 65.2%. These indexes decreased significantly from low to high urbanisation levels for both mosquito species. The relative abundance (RA) of Ae. aegypti immatures was slightly higher toward large cities, but showed no difference for Cx. pipiens. The CI of shaded tires was significantly higher than the CI of exposed tires for both mosquito species. There was no difference in RA values between shaded and sunlit tires. The CI and the SI were highest during the summer across the urbanisation levels, except for Cx. pipiens, which continued to increase during the autumn in small towns. Results related to urbanisation gradient, sunlit exposure and seasonality are discussed.

Identification of the natural breeding sites of sandflies (Diptera: Psychodidae: Phlebotominae), potential vectors of leishmaniasis, in the province of Chaco, Argentina

The aim of this work was to identify the natural breeding sites of sandflies in the province of Chaco, Argentina, for the first time. Preliminary studies were conducted in two different phytogeographic regions: dry Chaco (Parque Provincial Pampa del Indio), in January 2010, and humid Chaco (Resistencia, Margarita Belén and Colonia Benítez), from May-September 2010. A total of 127 samples were collected (Pampa del Indio: 15, Resistencia: 37, Margarita Belén: 36, Colonia Benítez: 39). A female of Migonemyia migonei was found in Pampa del Indio at the base of a bromeliad in the summer (January) and a pupal exuvium of a phlebotomine fly was found in Resistencia, in a place where dogs rested, in the winter (July). These findings highlighted these two sites as potential breeding sites. Because the existence of potential natural breeding sites for sandflies has been demonstrated in both forest and periurban areas, expanding the search efforts and characterising these sites will enable the development of specific study designs to gain insight into the spatial distribution of the risks posed by these vectors. The resulting information will serve as a basis for proposing and evaluating vector control measures.

New classification of natural breeding habitats for Neotropical anophelines in the Yanomami Indian Reserve, Amazon Region, Brazil and a new larval sampling methodology

Here we present the first in a series of articles about the ecology of immature stages of anophelines in the Brazilian Yanomami area. We propose a new larval habitat classification and a new larval sampling methodology. We also report some preliminary results illustrating the applicability of the methodology based on data collected in the Brazilian Amazon rainforest in a longitudinal study of two remote Yanomami communities, Parafuri and Toototobi. In these areas, we mapped and classified 112 natural breeding habitats located in low-order river systems based on their association with river flood pulses, seasonality and exposure to sun. Our classification rendered seven types of larval habitats: lakes associated with the river, which are subdivided into oxbow lakes and nonoxbow lakes, flooded areas associated with the river, flooded areas not associated with the river, rainfall pools, small forest streams, medium forest streams and rivers. The methodology for larval sampling was based on the accurate quantification of the effective breeding area, taking into account the area of the perimeter and subtypes of microenvironments present per larval habitat type using a laser range finder and a small portable inflatable boat. The new classification and new sampling methodology proposed herein may be useful in vector control programs.

Bonafide, type-specific human papillomavirus persistence among HIV-positive pregnant women: predictive value for cytological abnormalities, a longitudinal cohort study

This study investigated the rate of human papillomavirus (HPV) persistence, associated risk factors, and predictors of cytological alteration outcomes in a cohort of human immunodeficiency virus-infected pregnant women over an 18-month period. HPV was typed through L1 gene sequencing in cervical smears collected during gestation and at 12 months after delivery. Outcomes were defined as nonpersistence (clearance of the HPV in the 2nd sample), re-infection (detection of different types of HPV in the 2 samples), and type-specific HPV persistence (the same HPV type found in both samples). An unfavourable cytological outcome was considered when the second exam showed progression to squamous intraepithelial lesion or high squamous intraepithelial lesion. Ninety patients were studied. HPV DNA persistence occurred in 50% of the cases composed of type-specific persistence (30%) or re-infection (20%). A low CD4+T-cell count at entry was a risk factor for type-specific, re-infection, or HPV DNA persistence. The odds ratio (OR) was almost three times higher in the type-specific group when compared with the re-infection group (OR = 2.8; 95% confidence interval: 0.43-22.79). Our findings show that bonafide (type-specific) HPV persistence is a stronger predictor for the development of cytological abnormalities, highlighting the need for HPV typing as opposed to HPV DNA testing in the clinical setting.

Breeding sites of Neotropical Drosophilidae (Diptera): IV. living and fallen flowers of Sessea brasiliensis and Cestrum spp. (Solanaceae)

The flowering phenologies of Sessea brasiliensis and three species of Cestrum (C. amictum, C. schlechtendalii and C. sendtnerianum), all belonging to the Solanaceae, were monitored in a fragment of the montane Atlantic rain forest within São Paulo city from July 1993 through June 1995. A total of 11,021 flies belonging to five species of the flavopilosa group of Drosophila (D. cestri, D. cordeiroi, D. hollisae, D. incompta and D. mariaehelenae), plus 1,244 flies belonging to four species of Zygothrica (Z. dispar, Z. vittimaculosa, Z. vittinubila and Z. aff. vittipunctata) emerged from 33,955 sampled flowers. Cestrum amictum, C. sendtnerianum and Sessea brasiliensis are recorded for the first time as host plants for the species of the Neotropical flavopilosa group ofDrosophila. Additionally, at least ten less abundant taxa belonging also to the Drosophilidae, plus 1,073 microhymenopteran parasitoids (Braconidae, Eucoilidae, and Pteromalidae), emerged from the sampled flowers of S. brasiliensis e Cestrum spp. The present study raised from nine to eleven the number of species of Cestrum previously recorded as breeding sites for these flies. A comprehensive list of species of Solanaceae known to host six species of Drosophila and four species of Zygothrica in southeastern and southern Brazil is included.

Breeding sites of drosophilids (Diptera) in the Brazilian Savanna. I. Fallen fruits of Emmotum nitens (Icacinaceae), Hancornia speciosa (Apocynaceae) and Anacardium humile (Anacardiaceae)

In this study, drosophilids that bred in fruits of three common plant species of the Brazilian Savanna were investigated: Emmotum nitens, Hancornia speciosa and Anacardium humile, along with the temporal and spatial distribution of these insects among fruits obtained from six individuals of E. nitens. Fallen fruits were collected in natural environments, placed on moist sand in individual containers and all drosophilids that emerged from these resources during 15 days were collected, counted and identified. From 3,651 fruits collected (3,435 of E. nitens, 179 of A. humile and 37 of H. speciosa) 4,541 flies emerged and were classified into 19 species of Drosophilidae. Their distribution was unequal among the three resources, also over time, and among the six individuals of E. nitens. Such fluctuations probably reflect the availability of resources in time and space and probably the action of selective factors such as larval competition.