Identification and antimicrobial susceptibility of Staphylococcus from home-treated peritoneal dialysis patients

Staphylococcus aureus is the main agent of infections during peritoneal dialysis (PD). The presence of S. aureus in the nasal cavity has been extensively studied and suggested as a risk factor of dialysis-related infections, whereas coagulase-negative Staphylococcus (CNS) species are frequently considered part of the normal human microbiota. The aim of this study was to identify Staphylococcus in the nasal cavity, pericatheter skin and peritoneal effluent from PD patients, as well as to evaluate the antimicrobial activity evolution in vitro. Thirty-two chronic PD patients were observed during 12 months and had nasal and pericatheter skin samples collected for culture. When peritonitis was detected, samples were also collected from the peritoneal effluent for culture. The activity of several antimicrobial drugs (penicillin G, oxacillin, cephalothin, ofloxacin, netilmicin and vancomycin) against different Staphylococcus species was measured by using the agar drug diffusion assay (Kirby-Bauer method). Staphylococcus was separated into S. aureus, S. epidermidis and other CNS species in order to determine the in vitro resistance level. S. epidermidis resistance to oxacillin progressively increased during the study period (p < 0.05). Resistance to ofloxacin was inexpressive, whereas resistance to netilmicin and vancomycin was not detected. of the oxacillin-resistant species (n = 74), 83% were S. epidermidis, 13% other CNS and 4% S. aureus (p < 0.05). Regarding multidrug resistant strains (n = 45), 82% were S. epidermidis, 13% other CNS, and 5% S. aureus (p < 0.05). This study shows the relevance of resistance to oxacillin and CNS multi-drug resistance, particularly concerning S. epidermidis, in PD patients.

Molecular detection of enterotoxins E, G, H and I in Staphylococcus aureus and coagulase-negative staphylococci isolated from clinical samples of newborns in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Polymerase Chain Reaction Detection of Enterotoxins Genes in Coagulase-Negative Staphylococci Isolated from Brazilian Minas Cheese

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Kinetics of venom and antivenom serum levels, clinical evaluation and therapeutic effectiveness in dogs inoculated with Crotalus durissus terrificus venom

This work evaluated the clinical and therapeutic aspects as well as serum levels of venom and antivenom IgG by enzyme-linked immunosorbent assay (ELISA) in experimental envenomation of dogs with Crotalus durissus terrificus venom. Twenty-eight mixed breed adult dogs were divided into four groups of seven animals each, Group I: only venom; Group II, venom + 50 ml of anti-bothropic-crotalic serum (50mg) + fluid therapy; Group III, venom + 50 ml of anti-bothropic-crotalic serum + fluid therapy + urine alkalination; Group IV, 50 ml of anti-bothropic-crotalic serum. The lyophilized venom of Crotalus durissus terrificus was reconstituted in saline solution and subcutaneously inoculated at the dose of 1mg/kg body weight. The dogs presented clinical signs of local pain, weakness, mandibular ptosis, mydriasis, emesis and salivation. The venom levels detected by ELISA ranged from 0 to 90ng/ml, according to the severity of the clinical signs. Serum antivenom ranged from 0 to 3ug/ml and was detected for up to 138h after treatment. ELISA results showed the effectiveness of the serum therapy for the venom neutralization.

Checklist of helminths from lizards and amphisbaenians (Reptilia, Squamata) of South America

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Genotyping of Brazilian Giardia duodenalis human axenic isolates

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Taxa de desenvolvimento de Chrysomya albiceps(Wiedemann) (Diptera: Calliphoridae) em dieta artificial acrescida de tecido animal para uso forense

Forensic entomology uses biological and ecological aspects of necrophagous insects to help in criminal investigations to estimate the post-mortem interval (PMI) or to determine the cause of death. Recent papers demonstrated that the presence of toxins in decomposing tissues may alter the insect developmental rate of insects exploiting such tissues as food. Thus, preliminary tests with artificial diets in laboratory are necessary to create a database to investigate and quantify the modifications that can occur with the collected insects from a criminal scene, avoiding any errors on the PMI estimates. The present study aimed to evaluate the developmental rate of Chrysomya albiceps (Wiedemann) reared on: a) artificial diets containing animal tissues: bovine liver (D1), raw muscle (D2), stomach (D3), and chicken heart (D4); b) artificial diet without animal tissue (D5); and c) a control group (C), which had only meat. The efficiency of each substrate was assessed by immature weight gain (mg), larval developmental time, larval and pupal survival, emergence interval and adult size. D1 to D4 diets did not restrict C. albiceps development; however, larvae reared on D1 and D2 diets presented a lower adult emergence rate. D3 and control group showed similarities regarding the efficiency parameters (rate and emergence interval). Thus, the use of diet D3, artificial diet with stomach, is the most recommended.

New record of Haplometroides intercaecalis (Digenea, Plagiorchiidae) infecting a Brazilian snake

Phalotris matogrossensis (Serpentes, Colubridae) was reported as a new host for Haplometroides intercaecalis ( Digenea, Plagiorchiidae). The host snake was obtained from the municipality of Anastacio, state of Mato Grosso do Sul, Brazil. One specimen of H. intercaecalis was recovered from the esophagus of the host and identified by the intercecal position of the vitellaria in the pre-acetabular region. This paper describes the second report of the occurrence of this trematode in fossorial snakes of the genus Phalotris in the state of Mato Grosso do Sul, Brazil.

Diagnosis of Giardia infections by PCR-based methods in children of an endemic area

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Identification and characterization of excretory/secretory products of Giardia duodenalis trophozoites

One of the major questions concerning Giardia is the understanding of pathophysiological processes associated with small intestine abnormalities. There are evidences that Giardia trophozoites contain and/or release proteolytic enzymes that may be implicated in the host intestinal epithelium. The present investigation was undertaken to examine the protease activity in excretory/secretory (E/S) products of Giardia duodenalis trophozoites of an axenic Brazilian strain (BTU-11) and the reference strain Portland 1 (P1). E/S products from trophozoites of each strain in conditioned medium were tested with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for the protein profiles, and the protease activity was analyzed using substrate-impregnated SDS-PAGE (gelatin and collagen) and hemoglobin assay. The proteases characterization was based on inhibition assays including synthetic inhibitors. Electrophoresis analysis of E/S products revealed a banding pattern composed by few bands (4 to 6 bands) in the migration region of 123 to 28 kDa. Proteolytic products were detected in the conditioned medium by trophozoites of both assayed strains. In the gels containing copolymerized gelatin and collagen, E/S products promoted substrate degradation and the most evident proteolysis zones were distributed in the migration regions of 77 to 18 kDa and 145 to 18 kDa, respectively, in the patterns of gelatinolytic and collagenolytic activities. Degradation of hemoglobin was also observed, and the pattern of hydrolysis was similar in both E/S products assayed. Inhibitor assays showed that the main proteolytic activity in both E/S products is due to cysteine proteases, although the presence of serine proteases was also indicated. Degradation of substrates including collagen and hemoglobin could lead us to speculate different functions of Giardia excreted/secreted proteases in vivo, but to confirm this possibility and to elucidate its implication on host-parasite interactions, further experiments applying protocols for the purification of proteases are necessary. Even so, our observations are relevant and hold the perspective for the understanding about protease activity in Giardia trophozoites of axenic strain isolated in an endemic area.

New reports on parasitism by Haplometroides buccicola (Digenea, Plagiorchiidae) in Brazilian snakes

The occurrence of Haplometroides buccicola (Digenea, Plagiorchiidae) in the esophagus of two Brazilian snakes is reported in the present study. The trematodes were collected from one Micrurus corallinus (Elapidae) and one Phalotris lativittatus (Colubridae); both snakes were found in Botucatu city, São Paulo State, Brazil. Morphological and morphometric analyses of the trematodes are presented. For the first time Micrurus corallinus has been recorded as a host for H. buccicola and this is the second time that P. lativittatus has been reported as a host for this trematode species.

Acute neuromuscular manifestations in a patient associated with ingesting octopus (Octopus sp.)

Os autores relatam um quadro manifestado por sintomas neurológicos e musculares em uma mulher de 45 anos, que surgiu após o consumo da carne de polvo comum (Octopus sp.). A paciente apresentou intenso mal estar, parestesias em extremidades e área perioral, fraqueza muscular intensa e hipotensão arterial, seguidos de prurido importante e uma erupção eritêmato-descamativa disseminada tardia. Não foram observadas manifestações gastrintestinais ou febre, o que reduziu a probabilidade de uma intoxicação alimentar por conservação inadequada do molusco. A presença de sintomas neuro-musculares é sugestiva de ação de neurotoxinas, comprovadamente existentes em muitos gêneros de polvos e que podem ter sido ingeridas através do consumo das glândulas salivares ou acúmulo das toxinas na carne, por algum mecanismo ainda desconhecido. As toxinas dos polvos do gênero Octopus são pouco estudadas e julgamos esta comunicação importante por alertar para a possibilidade do envenenamento nos seres humanos que consomem carne de polvos e ainda sua diferenciação das intoxicações alimentares que ocorrem por conservação inadequada do animal.

Structure determination of a tetrahydro-beta-carboline of arthropod origin: A novel alkaloid-toxin subclass from the web of spider Nephila clavipes

The orb-web spiders are polyphagous animals in which the web plays a very important role in the capture of preys; oily droplets usually cover the capture-web of the spider Nephila clavipes and seem to be of great importance for prey capture. The knowledge of the chemical composition of these droplets is necessary to understand the function of this adhesive material in web mechanics and prey capture. A novel subclass of spider toxins, tetrahydro-beta-carboline, was identified among the weaponry of compounds present inside of oily droplets. This type of alkaloid is not common among the natural compounds of spider toxins. Apparently, when the prey arthropods get caught by the spider web, their bodies are covered with many adhesive oily droplets, which disrupt delivering the tetrahydro-beta-carboline to the direct contact with the prey integument. Toxicity assays demonstrated a potent lethal effect of the alkaloid toxin to the spider preys; topical applications of the teirahydro-beta-carboline at first caused clear signs of neurotoxicity, followed by the death of preys. The structure of the major component, a tetrahydro-beta-carboline, among the alkaloid toxins was elucidated by means of UV spectrophotometry, ESI mass spectrometry, H-1-NMR spectroscopy, and high-resolution mass spectrometry. The structure of the natural toxin was determined as 1-(2-guanidinoethyl)-1,2,3,4-tetrahydro-6-hydroxymethyl)-beta-carboline; the investigation of the pharmacological properties and neurotoxic actions of this compound may be used in the future as reference for the development of new drugs to be applied at level of pest control in agriculture.

Two new bradykinin-related peptides from the venom of the social wasp Protopolybia exigua (Saussure)

Two bradykinin-related peptides (Protopolybiakinin-I and Protopolybiakinin-II) were isolated from the venom of the social wasp Protopolybia exigua by RP-HPLC, and sequenced by Edman degradation method. Peptide sequences of Protopolybiakinin-I and Protopolybiakinin-II were DKNKKPIRVGGRRPPGFTR-OH and DKNKKPIWMAGFPGFTPIR-OH, respectively. Synthetic peptides with identical sequences to the bradykinin-related peptides and their biological functions were characterized. Protopolybiakinin-I caused less potent constriction of the isolated rat ileum muscles than bradykinin (BK). In addition, it caused degranulation of mast cells which was seven times more potent than BK. This peptide causes algesic effects due to the direct activation of B-2-receptors. Protopolybiakinin-II is not an agonist of rat ileum muscle and had no algesic effects. However, Protopolybiakinin-II was found to be 10 times more potent as a mast cell degranulator than BK. The amino acid sequence of Protopolybiakinin-I is the longest among the known wasp kinins. (c) 2006 Elsevier B.V. All rights reserved.

Insect Venom Peptides

The insects of the order Hymenoptera ( bees, wasps, and ants) are classified in two groups, based on their life history: social and solitary. The venoms of the social Hymenoptera evolved to be used as defensive tools to protect the colonies of these insects from the attacks of predators. Generally they do not cause lethal effects but cause mainly inflammatory and/or immunological reactions in the victims of their stings. However, sometimes it is also possible to observe the occurrence of systemic effects like respiratory and/or kidney failure. Meanwhile, the venoms of solitary Hymenoptera evolved mainly to cause paralysis of the preys in order to permit egg laying on/within the prey's body; thus, some components of these venoms cause permanent/transient paralysis in the preys, while other components seem to act preventing infections of the food and future progenies. The peptide components of venoms from Hymenoptera are spread over the molar mass range of 1400 to 7000 da and together comprise up to 70% of the weight of freeze-dried venoms. Most of these toxins are linear polycationic amphipatic peptides with a high content of alpha-helices in their secondary structures. These peptides generally account for cell lysis, hemolysis, antibiosis, and sometimes promote the delivery of cellular activators/mediators through interaction with the G-protein receptor, and perhaps some of them are even immunogenic components. In addition to these peptides, the Hymenopteran venoms also may contain a few neurotoxins that target Na+ and/or Ca+2 channels or even the nicotinic ACh receptor. This review summarizes current knowledge of the biologically active Hymenoptera venoms.