Prevalence of HIV type 1 drug resistance mutations in treatment-naïve and experienced patients from resource-limited settings with universal access to antiretroviral therapy: a survey in two small Brazilian cities

Concerns have been raised that universal availability of antiretroviral agents in resource-limited settings might lead to the emergence and spread of resistant strains. We present the largest survey on human immunodeficiency virus type 1 (HIV-1) resistance among treatment-naïve and experienced patients followed in small, relatively underprivileged cities in Brazil with universal availability to standard of care antiretroviral combinations. Samples were collected between 2004 and 2006 from 95 patients followed in the cities of Saquarema and Santo Antonio de Pádua, state of Rio de Janeiro. A proviral fragment encompassing protease and reverse transcriptase (RT) regions was generated and drug susceptibility level was inferred. Among 50 strains from drug-naïve subjects, one (2%) had intermediate-level resistance to RT inhibitors. Among 38 patients on therapy as of sampling, 28 (73.7%) had plasma viral load (PVL) below detection limit (26 of whom without evidence of resistance mutations) and 11 (28.9%) harbored strains with reduced susceptibility. Only two strains harbored both protease and RT inhibitor mutations. Among seven patients who were off-treatment as of sampling, two (28.5%) harbored strains with reduced susceptibility to RT inhibitors. The relatively high frequency of undetectable PVL among patients on treatment and the overall low prevalence of resistance-associated mutations are reassuring. Continued surveillance, however, is necessary.

Protective immunity against Trypanosoma cruzi

Upon infection, Trypanosoma cruzi triggers a strong immune response that has both protective and pathological consequences. In this work, several important questions regarding protective immunity are reviewed. Emphasis is placed on recent studies of the important protective role of CD8+ T cells and on previous studies of immunisation of domestic T. cruzi reservoirs that sought to address practical vaccination problems. Research on the maturation of memory cells and studies indicating that the prevalence of T. cruzi-specific T-cell responses and a high frequency of committed CD8+ T cells are associated with better clinical outcomes are also reviewed. Additionally, animal models in which protection was achieved without immunopathological consequences are discussed.

Prevalence and genotyping of hepatitis C virus in blood donors in the state of Pará, Northern Brazil

Given the scarcity of epidemiological information on hepatitis C virus (HCV) infection in Northern Brazil, we determined the prevalence and genotypic frequency in blood donors in the state of Pará (PA). Blood samples from all of the blood donors at the Fundação HEMOPA (blood bank of PA) from 2004-2006 were screened for the presence of antibodies to anti-HCV and samples seroreactive to anti-HCV were further tested for HCV RNA using real-time PCR. In total, 116 HCV-RNA samples were genotyped, based on maximum likelihood phylogenetic analyses, using BioEdit, Modelgenerator, PHYML and FigTree software. The population consisted of 242,726 volunteers who donated blood from 2004-2006; the most common subgroup was males between the ages of 18-29 years old (37.30%). Within the whole group, 1,112 blood donors (0.46%) had indeterminate or positive serology; among these, 28.78% were males whose ages ranged from 18-29 years. A diagnosis of chronic HCV infection was confirmed for 304 donors (60.20% males; 66.45% were 30-49 years old), resulting in a prevalence of HCV RNA in 0.13% of the samples (304 of 242,726). HCV genotyping revealed a high frequency of genotype 1 (108/116) followed by genotype 3 (8/116). This study found HCV infection to be relatively infrequent in PA; genotype 1 was most commonly isolated. This information can help guide prevention and control policies aimed at efficient diagnosis and control measures.

Evaluation of the synergistic potential of vancomycin combined with other antimicrobial agents against methicillin-resistant Staphylococcus aureus and coagulase-negative Staphylococcus spp strains

Methicillin-resistant Staphylococcus aureus (MRSA) and coagulase-negative Staphylococcus spp (CNS) are the most common pathogens that cause serious long term infections in patients. Despite the existence of new antimicrobial agents, such as linezolid, vancomycin (VAN) remains the standard therapy for the treatment of infections caused by these multidrug-resistant strains. However, the use of VAN has been associated with a high frequency of therapeutic failures in some clinical scenarios, mainly with decreasing concentration of VAN. This work aims to evaluate the synergic potential of VAN plus sulfamethoxazole/trimethoprim (SXT), VAN plus rifampin (RIF) and VAN plus imipenem (IPM) in sub-minimum inhibitory concentrations against 22 clinical strains of MRSA and CNS. The checkerboard method showed synergism of VAN/RIF and VAN/SXT against two and three of the 22 strains, respectively. The combination of VAN with IPM showed synergistic effects against 21 out of 22 strains by the E-test method. Four strains were analyzed by the time-kill curve method and synergistic activity was observed with VAN/SXT, VAN/RIF and especially VAN/IPM in sub-inhibitory concentrations. It would be interesting to determine if synergy occurs in vivo. Evidence of in vivo synergy could lead to a reduction of the standard VAN dosage or treatment time.

Sequence analysis of coding DNA fragments of pfcrt and pfmdr-1 genes in Plasmodium falciparum isolates from Odisha, India

The global emergence and spread of malaria parasites resistant to antimalarial drugs is the major problem in malaria control. The genetic basis of the parasite's resistance to the antimalarial drug chloroquine (CQ) is well-documented, allowing for the analysis of field isolates of malaria parasites to address evolutionary questions concerning the origin and spread of CQ-resistance. Here, we present DNA sequence analyses of both the second exon of the Plasmodium falciparum CQ-resistance transporter (pfcrt) gene and the 5' end of the P. falciparum multidrug-resistance 1 (pfmdr-1) gene in 40 P. falciparum field isolates collected from eight different localities of Odisha, India. First, we genotyped the samples for the pfcrt K76T and pfmdr-1 N86Y mutations in these two genes, which are the mutations primarily implicated in CQ-resistance. We further analyzed amino acid changes in codons 72-76 of the pfcrt haplotypes. Interestingly, both the K76T and N86Y mutations were found to co-exist in 32 out of the total 40 isolates, which were of either the CVIET or SVMNT haplotype, while the remaining eight isolates were of the CVMNK haplotype. In total, eight nonsynonymous single nucleotide polymorphisms (SNPs) were observed, six in the pfcrt gene and two in the pfmdr-1 gene. One poorly studied SNP in the pfcrt gene (A97T) was found at a high frequency in many P. falciparum samples. Using population genetics to analyze these two gene fragments, we revealed comparatively higher nucleotide diversity in the pfcrt gene than in the pfmdr-1 gene. Furthermore, linkage disequilibrium was found to be tight between closely spaced SNPs of the pfcrt gene. Finally, both the pfcrt and the pfmdr-1 genes were found to evolve under the standard neutral model of molecular evolution.

Elimination of Rhodnius prolixus in El Salvador, Central America

Epidemiological studies performed in El Salvador between 1955-1972 demonstrated a high frequency of two triatomine vectors (Triatoma dimidiata and Rhodnius prolixus). However, the frequency of R. prolixus decreased by 1976 and this species has not been found since 1995. The main factors influencing the elimination of R. prolixus in El Salvador are discussed in this paper.

Plasmodium simium/Plasmodium vivax infections in southern brown howler monkeys from the Atlantic Forest

Blood infection by the simian parasite, Plasmodium simium, was identified in captive (n = 45, 4.4%) and in wild Alouatta clamitans monkeys (n = 20, 35%) from the Atlantic Forest of southern Brazil. A single malaria infection was symptomatic and the monkey presented clinical and haematological alterations. A high frequency of Plasmodium vivax-specific antibodies was detected among these monkeys, with 87% of the monkeys testing positive against P. vivax antigens. These findings highlight the possibility of malaria as a zoonosis in the remaining Atlantic Forest and its impact on the epidemiology of the disease.

Association between perineal trauma and pain in primiparous women

Objective To identify the association between perineal trauma and pain in 473 primiparous women. Method Cross-sectional study in which pain was measured by the numerical pain scale (0 to 10 - 0 being no pain and 10 maximal pain). Results The prevalence and mean intensity of pain were 33.0% and 4.7 points (standard deviation = 2.0) in the numeric scale, respectively. Episiotomy represented the most frequent trauma (46.7%). The occurrence and intensity of the pain were associated with perineal trauma and postpartum time. Having perineal trauma tripled the chance of pain. Each hour elapsed following the birth reduced the chance of pain by 4.8%. Conclusion Primiparous women are subject to a high frequency of perineal trauma, with episiotomy being the most prominent. Perineal pain affects approximately one-third of primiparous women and is associated with the postpartum time and perineal traumas.

Rate of development of forensically-important Dipterain southern Brazil

Dipteran larvae were collected from rabbit (Oryctolagus cunniculus L.) carcasses during the four seasons in 2005 in the southernmost state of Rio Grande do Sul, Brazil. The larvae were fed ground beef at ambient temperatures following collection from carcasses. The development of each species under these conditions was estimated. The most abundant species in the carcasses were Lucilia eximia (Wiedemann) and Chrysomya albiceps (Wiedemann) (Calliphoridae), and they were found in all seasons. The data were fitted to a linear model that describes the relationship between temperature and linear developmental rating. These two species are primary forensic indicators in southern Brazil. Other species such as Hemilucilia semidiaphana (Rondani) (Calliphoridae), Synthesiomyia nudiseta (Wulp), Muscina stabulans (Fallen) (Muscidae), and Fannia pusio (Wiedemann) (Fanniidae) were forensically less important because they only occurred in high frequency in certain seasons and during the first days of carcass decomposition.

Nesting and use of pollen resources by Tetrapedia diversipes Klug (Apidae) in Atlantic Forest areas (Rio de Janeiro, Brazil) in different stages of regeneration

Nesting and use of pollen resources by Tetrapedia diversipes Klug (Apidae) in Atlantic Forest areas (Rio de Janeiro, Brazil) in different stages of regeneration. The nesting in trap-nests and use of pollen sources in larval food by Tetrapedia diversipes Klug, 1810 (Apidae) was compared between regenerating areas of Atlantic Forest. The study was conducted between April 2008 and October 2009 at União Biological Reserve, Rio de Janeiro, Brazil. T. diversipes nested in 66 trap-nests and showed a peak of nesting during the months of highest rainfall. The most frequent pollen type in brood cells during the wet season was Dalechampia sp. 1. During the dry season, the type Ludwigia sp. was the most frequent, followed by Dalechampia sp. 2. The high frequency of Dalechampia and Ludwigia species in the larval food, observed in both habitats and in the two seasons could be considered relevant for T. diversipes, suggesting highly selective diet based primarily on two plant species unrelated, but similar in size of pollen grains.

Community structures of soil animals and survival of land snails on an island of the Ogasawara Archipelago

On Chichijima, one of the Ogasawara (Bonin) Islands located in the Western Pacific Ocean, land snails have declined, the suggested cause being predation pressure by an invasive flatworm (Platydemus manokwari). Soil fauna were investigated in areas where the snail survives, and where it has become extinct. Much of the fauna, dominated by introduced earthworms and ants, was undiminished, however, one undescribed but endemic carabid (Badister sp.), which selectively feeds on land snails, was absent in snail-extinct areas. The invasive flatworm P. manokwari has been reported to feed also on the carcasses of earthworms, as well as on live snails, and is therefore expected to occur in most parts of Chichijima Island. Among other groups, the density of isopods (also dominated by exotic species) was very low, in comparison with the reported ones 30 years ago. Community structure is currently reflected by dominance of earthworms and ants, decline of endemic isopods, and a high frequency of introduced or alien species.

Plasmid DNA damage induced by singlet molecular oxygen released from the naphthalene endoperoxide DHPNO2 and photoactivated methylene blue

To investigate oxidative lesions and strand breaks induction by singlet molecular oxygen (¹O2), supercoiled-DNA plasmid was treated with thermo-dissociated DHPNO2 and photoactivated-methylene blue. DNA lesions were detected by Fpg that cleaves DNA at certain oxidized bases, and T4-endoV, which cleaves DNA at cyclobutane pyrimidine dimers and apurinic/apyrimidinic (AP) sites. These cleavages form open relaxed-DNA structures, which are discriminated from supercoiled-DNA. DHPNO2 or photoactivated-MB treatments result in similar plasmid damage profile: low number of single-strand breaks or AP-sites and high frequency of Fpg-sensitive sites; confirming that base oxidation is the main product for both reactions and that ¹O2 might be the most likely intermediate that reacts with DNA.

Ocorrência de Pratylenchus brachyurus e Meloidogyne incognita na cultura do algodoeiro no Estado do Mato Grosso

In a survey carried out in the state of Mato Grosso, Brazil, the lesion nematode, Pratylenchus brachyurus, and the root-knot nematode, Meloidogyne incognita, were found, respectively, in 94% and 3.7% of a total of 623 root and soil samples representing 21,793 ha. No visible aboveground symptoms in cotton (Gossypium hirsutum) plants infected by P. brachyurus were observed, except for typical root lesions. In contrast, plants with M. incognita usually exhibited pronounced symptoms of damage. The high frequency (94%) of P. brachyurus was unexpected and is of concern considering the fact that soybean (Glycine max) and corn (Zea mays), are both susceptible to this nematode and are used in crop rotation programs.

Water storage in wetted strips under irrigated coffee trees with different criteria of irrigation management

The increasing demand for water resources accentuates the need to reduce water waste through a more appropriate irrigation management. In the particular case of irrigated coffee planting, which in recent years presented growth with the predominance of drip irrigation, the improvement of drip irrigation management techniques is a necessity. The proper management of drip irrigation depends on the knowledge of the spatial pattern of soil moisture distribution inside the wetted strip formed under the irrigation lines. In this study, grids of 24 tensiometers were used to determine the water storage within the wetted strip formed under drippers, with a 3.78 L h-1 discharge, evenly spaced by 0.4 m, subjected to two different management criteria (fixed irrigation interval and 60 kPa tension). Estimates of storage based on a one-dimensional analysis, that only considers depth variations, were compared with two-dimensional estimates. The results indicate that for high-frequency irrigation the one-dimensional analysis is not appropriate. However, under less frequent irrigation, the two-dimensional analysis is dispensable, being the one-dimensional sufficient for calculating the water volume stored in the wetted strip.

Anaplasma marginale infection in cattle from south-western Amazonia

The present study provides the first epidemiological data regarding infection by Anaplasma marginale in cattle reared in south-western Brazilian Amazonia. One simple procedure was adapted for the extraction of DNA from blood clots collected in seven microregions of Rondônia State and two mesoregions of Acre State. PCR method was used to asses the frequency of A. marginale infections in 4 to12-month-old cattle. The cattle infection was investigated by polymerase chain reaction (PCR) using the specific primer "msp5" for A. marginale. The DNA amplifications revealed that the mean frequency of A. marginale infection was 98.6% (1,627/1,650) in samples from Rondonia, and 92.87% (208/225) in samples from Acre. The high frequency of A. marginale infections in 4 to 12-month-old cattle indicate a situation of enzootic stability in the studied areas and are comparable to those detected by immunodiagnosis in different endemic regions in Brazil. The DNA extraction of clotted blood method described here can be used for epidemiological studies on anaplasmosis and other bovine hemoparasites.