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Purpose. Chemokines are important in the immune response against viral infections, and may play a role in human T-lymphotropic virus 1 (HTLV-1)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) pathogenesis. Polymorphisms in the Duffy antigen receptor for chemokines (DARC), such as rs12075 (A>G; FY*B>FY*A ) and rs281477 (-46T>C; GATA-1 box) may influence circulating concentrations of proinflammatory chemokines. We investigate whether Duffy genotypes influence the HTLV-1 proviral load (PVL) level, HTLV-1 infection outcome and chemokine concentrations in HTLV-1 asymptomatic carriers (AC=162), HAM/TSP patients (HAM=135) and seronegative individuals (SN=71). Methodology. Quantification of plasmatic IL8, CCL2 and CCL5 were performed by flow cytometry and Duffy genotypes were investigated by real-time PCR. HTLV-1 PVL was quantified in peripheral blood. To control for spurious association, individual ancestry profiles in AC and HAM groups were investigated. Results/Key findings. PVL and IL8 level were significantly higher in the HAM group than in the AC group, but were not associated with Duffy genotypes. The highest CCL2 and CCL5 levels were seen in the SN group, and there was no difference when comparing the infected groups. The level of CCL5 was not associated with Duffy genotypes. The polymorphism -46 C/C that abrogates the DARC expression on the erythrocytes was significantly associated with lower levels of CCL2, neutrophil and white blood cell (WBC) counts in HTLV-1-infected individuals. Conclusion. We conclude that although the Duffy null genotype was associated with leukopenia, neutropenia and lower levels of CCL2, the data do not suggest the influence of Duffy genotypes on the neurologic outcome of HTLV-1 infection, but may be a confounding factor in comparison HTLV-1-infected populations with different ancestries, especially when defining inflammatory biomarkers.

Since potential changes in the dynamics and mobility of drugs upon complexation for delivery may affect their ultimate efficacy, we have investigated the dynamics of two local anesthetic molecules, bupivacaine (BVC, C 18 H 28 N 2 O) and ropivacaine (RVC, C 17 H 26 N 2 O), in both their crystalline forms and complexed with water-soluble oligosaccharide 2-hydroxypropyl-β-cyclodextrin (HP-β-CD). The study was carried out by neutron scattering spectroscopy, along with thermal analysis, and density functional theory computation. Mean square displacements suggest that RVC may be less flexible in crystalline form than BVC, but both molecules exhibit very similar dynamics when confined in HP-β-CD. The use of vibrational analysis by density functional theory (DFT) made possible the identification of molecular modes that are most affected in both molecules by insertion into HP-β-CD, namely those of the piperidine rings and methyl groups. Nonetheless, the somewhat greater structure in the vibrational spectrum at room temperature of complexed RVC than that of BVC, suggests that the effects of complexation are more severe for the latter. This unique approach to the molecular level study of encapsulated drugs should lead to deeper understanding of their mobility and the respective release dynamics., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Although the presence of endosymbiotic rickettsial bacteria, specifically Candidatus Megaira, has been reported in diverse habitats and a wide range of eukaryotic hosts, it remains unclear how broadly Ca. Megaira are distributed in a single host species. In this study we seek to address whether Ca. Megaira are present in most, if not all isolates, of the parasitic ciliate Ichthyophthirius multifiliis . Conserved regions of bacterial 16S rRNA genes were either PCR amplified, or assembled from deep sequencing data, from 18 isolates/populations of I. multifiliis sampled worldwide (Brazil, Taiwan, and USA). We found that rickettsial rRNA sequences belonging to three out of four Ca. Megaira subclades could be consistently detected in all I. multifiliis samples. I. multifiliis collected from local fish farms tend to be inhabited by the same subclade of Ca. Megaira, whereas those derived from pet fish are often inhabited by more than one subclade of Ca. Megaira. Distributions of Ca. Megaira in I. multifiliis thus better reflect the travel history, but not the phylogeny, of I. multifiliis . In summary, our results suggest that I. multifiliis may be dependent on this endosymbiotic relationship, and the association between Ca. Megaira and I. multifiliis is more diverse than previously thought.

HTLV-1 proviral load (pvl) is an important risk marker for HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP), but its value as prognostic marker is not well defined. Long-term prospective cohort studies are necessary to clarify this question. Here, we analyzed HTLV-1 pvl in the peripheral blood of 82 asymptomatic carriers (AC; 351 samples), 12 HAM/TSP patients (HAM; 46 samples), and six incident cases of HAM/TSP (iHAM), with serial samples collected before (n = 10) and after (n = 20) the disease onset. The mean interval of follow-up was 10 years in the AC group and 8 years in HAM and iHAM groups. pvl was not significantly different between the first and last measurements in the three groups, but there was a trend to decrease over time. Coefficient of variation of pvl was significantly lower in the AC group than in HAM (p = 0.015) and iHAM (p = 0.022) patients. AC and HAM individuals showed a significant and strong positive correlation between the first and last measurements of pvl, but not iHAM subjects. All individuals who developed HAM/TSP during the follow-up had high pvl level (>1 %) before the onset of disease, but a typical increase in pvl was not observed in that period. The data suggest that there is a trend to reach an equilibrium plateau of pvl over time, characteristic of each individual. A significant rate of AC keeps high pvl levels for a long time without developing clinical symptoms associated to HTLV-1 infection. Thus, serial quantification of pvl in the peripheral blood does not seem to be a good prognostic marker for HAM/TSP.

Pseudocapillaria tomentosa is a common pathogen of zebrafish (Danio rerio) in research facilities. We developed a method to collect and concentrate the nematode eggs using a modified sugar centrifugation method and documented their normal development. Embryonating stages with blastomere formation followed by elongation of the embryo prior to larva formation cumulated in developed larvae inside the eggs and hatching after 5-10 day. We then evaluated the efficacy of heat and chlorine to kill them based on a larva development assay. Eggs were exposed to 40, 50, 60 °C for 30 min and 1 h. Chlorine treatment was performed at 100, 250, 500, 1000, 3000 and 6000 ppm for 10 min. Samples exposed to 40 °C for 30 min or 1 h showed incidences of larvated eggs similar to controls. In contrast, no larvation occurred with eggs exposed to either 50 or 60 °C for 30 min or 1 h. Remarkably, in repeated assays, samples exposed to low doses of chlorine (100, 250, 500 and 1000 ppm for 10 min) showed significantly higher incidence of larvation than controls. Eggs treated with 3000 ppm for 10 min did not develop larvae, and no eggs were found after 6000 ppm treatment., (© 2016 John Wiley & Sons Ltd.)

Parasitic isopods of fishes usually show wide distribution and low host specificity. This study investigated the occurrence of gill parasites in 120 specimens of spotted goatfish Pseudupeneus maculatus, marine fish of great economic importance for fishery community. The fish were captured monthly in the Coast of Pernambuco State, Northeast Brazil, between October 2012 and September 2013. Eleven isopods were found and identified as Rocinela signata. We observed lower infections in the gills of P. maculatus (prevalence 8.3%, mean intensity 1.6±0.3 and mean abundance 0.1±0.3) when compared to other studies of different host fishes from geographically close locations. This parasite has been reported from a wide variety of fish species, not only in Brazil, but also in the eastern Atlantic and the Pacific oceans. This is the first report of R. signata in spotted goatfish.

The present study describes a new species of Trichodina found in the mucus of the body surface of 137 farmed and wild silver catfish (Rhamdia quelen) located in southern Brazil and in the gills of 99 banded knife fish (Gymnotus sp.) captured in the basin of Pantanal. Smears of the body surface mucus and gills were air dried at room temperature and impregnated with silver nitrate and stained with Giemsa for analysis of the adhesive disc, denticles and macronucleus. Characterized as a medium trichodinid, Trichodina quelenii n. sp. resembles T. heterodentata and T. acuta. It differs from T. heterodentata by the shape of the blade and from T. acuta by the lack of central circle. Furthermore, the distinct shape of the blade of T. quelenii n. sp. differs from all previously described species for the genus, leaving no doubt it is a new species.

Objective: To verify genetic determinants associated with stroke in children with sickle cell disease (SCD)., Methods: Prospective cohort with 110 children submitted to neonatal screening by the Neonatal Screening Program, between 1998 and 2007, with SCD diagnosis, followed at a regional reference public service for hemoglobinopathies. The analyzed variables were type of hemoglobinopathy, gender, coexistence with alpha thalassemia (α-thal), haplotypes of the beta globin chain cluster, and stroke. The final analysis was conducted with 66 children with sickle cell anemia (SCA), using the chi-squared test in the program SPSS ® version 14.0., Results: Among children with SCD, 60% had SCA. The prevalence of coexistence with α-thal was 30.3% and the Bantu haplotype (CAR) was identified in 89.2%. The incidence of stroke was significantly higher in those with SCA (27.3% vs. 2.3%; p=0.001) and males (24.1% vs. 9.6%; p=0.044). The presence of α-thal (p=0.196), the CAR haplotype (p=0.543), and socioeconomic factors were not statistically significant in association with the occurrence of stroke., Conclusion: There is a high incidence of stroke in male children and in children with SCA. Coexistence with α-thal and haplotypes of the beta globin chain cluster did not show any significant association with stroke. The heterogeneity between previously evaluated populations, the non-reproducibility between studies, and the need to identify factors associated with stroke in patients with SCA indicate the necessity of conducting further research to demonstrate the relevance of genetic factors in stroke related to SCD., (Copyright © 2016 Sociedade Brasileira de Pediatria. Published by Elsevier Editora Ltda. All rights reserved.)

Most of the analytical methods currently applied in food control laboratories are focused on the determination of target compounds using LC coupled to tandem MS, which is an effective technique, but low-resolution MS is limited. Thus, a method for determination of pesticide multiresidues in fruits (pear, apple, and grape) using a modified quick, easy, cheap, effective, rugged, and safe method and LC coupled to quadrupole time-of-flight (Q-TOF) MS was developed and validated. The proposed method showed good linearity (r2 > 0.99) from 1 to 100 μg/L. Recoveries for blank samples spiked at 0.01, 0.04, and 0.10 mg/kg were between 66 and 122%, with RSDs <28%. Respective LOQs for apple, pear, and grape matrixes were 0.01 mg/kg for 112, 120, and 118 compounds, and 0.04 mg/kg for 22, 12, and 17 compounds, and average mass accuracy error was 3.2 ppm. LC with Q-TOF MS detection using protonated molecular ion and/or adducts and mass accuracy provided reliability for the method. The proposed method is effective for pesticide residue determination in apple, pear, and grape samples, proving that high-resolution MS using full scan mode can be a powerful and reliable technique for quantification purposes, being adequate for application in the surveillance of maximum residue limits set by different legislations.

Objective:: This study aimed to evaluate the effectiveness of two methods of visual magnification (operating microscope and light head magnifying glass) for removal of composite flash around orthodontic metal brackets., Material and Methods:: Brackets were bonded in the center of the clinical crown of sixty well-preserved human premolars. Half of the sample was bonded with conventional Transbond XT (3M Unitek TM, USA), whereas the other half was bonded with Transbond TM Plus Color Change (3M Unitek TM, USA). For each type of composite, the choice of method to remove the flash was determined by randomly distributing the teeth into the following subgroups: A (removal by naked eye, n = 10), B (removal with the aid of light head magnifying glass, under 4x magnification, n = 10), and C (removal with the aid of an operating microscope, under 40x magnification, n = 10). Brackets were debonded and teeth taken to a scanning electron microscope (SS-x-550, Shimadzu, Japan) for visualization of their buccal surface. Quantification of composite flash was performed with Image Pro Plus software, and values were compared by Kruskal-Wallis test and Dunn's post-hoc test at 5% significance level., Results:: Removal of pigmented orthodontic adhesive with the aid of light head magnifying glass proved, in general, to be advantageous in comparison to all other methods., Conclusion:: There was no advantage in using Transbond TM Plus Color Change alone. Further studies are necessary to draw a more definitive conclusion in regards to the benefits of using an operating microscope.

The tasks of caregiving for children with disabilities involve contextual life stressors. The study aimed to investigate the coping strategies used by parents of children with intellectual disabilities (ID). The search included articles published between 2001 and 2015 on the PubMed, LILACS and PsycINFO databases, using the following key words: intellectual disability, coping, parents, caregivers, family. Thirteen articles were selected, the majority of which adopt a cross-sectional, comparative and quantitative approach. Results indicated that caring for a person with ID is stressful for the family. Parental stress generates changes in the family dynamic and coping strategies are used to facilitate the interaction of the caregiver with the person with ID. Seeking social support in family and professional services, the union of spouses, and search for information, among others, are the main strategies used. Social support was strongly associated with strengthening of the family unit. In contrast, wishful thinking, self-blame, distancing and social isolation were negatively associated with family unity. It is necessary to sensitize health professionals and educators to embrace the theme in public policies and preventive, diagnostic and therapeutic strategies.

This study evaluated the use of sugarcane filter cake and nitrogen, phosphorus and potassium (NPK) fertilization in the bioremediation of a soil contaminated with diesel fuel using a completely randomized design. Five treatments (uncontaminated soil, T1; soil contaminated with diesel, T2; soil contaminated with diesel and treated with 15 % (wt) filter cake, T3; soil contaminated with diesel and treated with NPK fertilizer, T4; and soil contaminated with diesel and treated with 15 % (wt) filter cake and NPK fertilizer, T5) and four evaluation periods (1, 60, 120, and 180 days after the beginning of the experiment) were used according to a 4 × 5 factorial design to analyze CO2 release. The variables total organic carbon (TOC) and total petroleum hydrocarbons (TPH) remaining in the soil were analyzed using a 5 × 2 factorial design, with the same treatments described above and two evaluation periods (1 and 180 days after the beginning of the experiment). In T3 and T5, CO2 release was significantly higher, compared with the other treatments. Significant TPH removal was observed on day 180, when percent removal values were 61.9, 70.1, 68.2, and 75.9 in treatments T2, T3, T4, and T5, respectively, compared with the initial value (T1).

Among calibration approaches for organic compounds determination in complex matrices, external calibration, based in solutions of the analytes in solvent or in blank matrix extracts, is the most applied approach. Although matrix matched calibration (MMC) can compensates the matrix effects, it does not compensate low recovery results. In this way, standard addition (SA) and procedural standard calibration (PSC) are usual alternatives, despite they generate more sample and/or matrix blanks consumption need, extra sample preparations and higher time and costs. Thus, the goal of this work was to establish a fast and efficient calibration approach, the diluted standard addition calibration (DSAC), based on successive dilutions of a spiked blank sample. In order to evaluate the proposed approach, solvent calibration (SC), MMC, PSC and DSAC were applied to evaluate recovery results of grape blank samples spiked with 66 pesticides. Samples were extracted with the acetate QuEChERS method and the compounds determined by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). Results indicated that low recovery results for some pesticides were compensated by both PSC and DSAC approaches. Considering recoveries from 70 to 120% with RSD <20% as adequate, DSAC presented 83%, 98% and 100% of compounds meeting this criteria for the spiking levels 10, 50 and 100μgkg(-1), respectively. PSC presented same results (83%, 98% and 100%), better than those obtained by MMC (79%, 95% and 97%) and by SC (62%, 70% and 79%). The DSAC strategy showed to be suitable for calibration of multiresidue determination methods, producing adequate results in terms of trueness and is easier and faster to perform than other approaches., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The human T-cell leukemia virus type 1 (HTLV-1) is present throughout the world and is associated with HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and other inflammatory conditions. The pathogenesis of HAM/TSP involves a chronic inflammatory response in central nervous system (CNS), with the presence of HTLV-1 infected cells and HTLV-1-specific CD8+ lymphocytes. Chemokines may have a role in the infiltration of these cells into the CNS. In this context, the present study analyzed the level of plasmatic chemokines CCL2 (MCP-1), CCL5 (RANTES), IL8 (CXCL8), CXCL9 (MIG), and CXCL10 (IP-10) and HTLV-1 proviral load from peripheral blood in 162 asymptomatic carriers and 136 HAM/TSP patients to determine the differences that be associated with the clinical status of the HTLV-1 infection. The results showed that patients with HAM/TSP have significantly higher levels of IL8 and CXCL9, and that the level of IL8, CXCL9 and CXCL10 was significantly greater in HTLV-1 infected individuals with high (>1%) than those with low proviral load (<1%). However, the levels of the chemokines tested have not showed high sensitivity to discriminate HAM/TSP patients from asymptomatic carriers. In addition, chemokine profiles in asymptomatic carriers and HAM/TSP groups were similar, with no significant increased frequency of higher producers of chemokines in HAM/TSP individuals. Results indicate that the heterogeneity of the individuals in the groups regarding time of infection, duration of disease, proviral load level and other possible confound factors may impair the use of chemokines levels to monitor HTLV-1 carriers in clinical practice. J. Med. Virol. 88:1438-1447, 2016. © 2016 Wiley Periodicals, Inc., (© 2016 Wiley Periodicals, Inc.)

This study evaluated the parasite fauna of farmed hybrid surubim (Pseudoplatystoma reticulatum x P. corruscans) and the host-parasite-environment relationship in two fish farms located in Mato Grosso do Sul, Central Brazil, South America. A total of 120 hybrids from two different farms, 60 in each season (30 in the hot and 30 in cold season) were examined during a year. Water quality was weekly measured to evaluate the interaction among environmental conditions and parasitism. Histopathology was used to observe the effects of the parasites and environment on the fish gills. The ciliate protozoan Ichthyophthirius multifiliis and the monogeneans (Ameloblastella sp., Amphocleithrium paraguayensis, Vancleaveus ciccinus, V. fungulus and V. janacauensis) were the most prevalent parasites detected in both seasons in both farms, with prevalence above 80%. It was stated that parasites did not cause important damage in the health status of the hybrid surubim. These results might be related to general good management practices and environmental quality implemented by the fish farmers. The presence of uncommon monogenean parasites to this hybrid compared to their parents causing an environmental and ecological concern is here discussed.

This study characterizes by transmission electron microscopy (TEM) and morphometric features the myxozoan Myxobolus cuneus (Myxosporea) in Piaractus mesopotamicus and reports the skeletal muscle and kidney as site of infection. The register was based in 21 young fish from intensive fish farming in Southeast Brazil and the spores were analyzed in fresh-mounted slides of the infected organs stained with Toluidine blue and processed as usual for TEM. It differs from Myxobolus cunhai from the fish host and different polar capsule size, and from Myxobolus serrasalmi on the pyriform spore shape and an oval macrospore, differently to that reported in this study. Morphometric characteristics and TEM study confirmed the present material as M. cuneus.

A new species of Tripartiella is described from the gills of the wild saddle cichlid Aequidens tetramerus in north Brazil. Wet smears of skin and gills of examined fish were air-dried at room temperature and impregnated with Klein's dry silver method for examination of the adhesive disc's structures and denticles. Total prevalence of parasitism was 65%. This ciliate is characterized as a small-sized trichodinid, body diameter 37.03 ± 4.9 μm, adhesive disc 30.50 ± 2.71 μm, denticulate ring 13.28 ± 0.8 μm and 24 ± 2.0 denticles. Taxonomic and morphometric data for the new species are discussed.

The tumor necrosis factor like weak inducer of apoptosis (TWEAK) and its receptor, fibroblast growth factor-inducible 14 (Fn14), mediate inflammation and neuronal apoptosis in cerebral edema, ischemic stroke and multiple sclerosis. The downstream effectors and pathways linked to TWEAK-Fn14 signaling are strongly implicated in the pathology of Parkinson's disease (PD), thus indicating a putative role for TWEAK/Fn14 signaling in PD neurodegeneration. Using the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model, we aimed to determine whether genetic ablation or pharmacologic mitigation of the TWEAK protein and its Fn14 receptor affected substantia nigra and striatum Parkinsonian pathology. Changes in endogenous TWEAK protein expression were also quantified in tissue from both MPTP-treated mice and PD human samples. TWEAK protein expression was transiently increased in the striatal tissue but remained unaltered in substantia nigra tissue of MPTP-treated mice. There was also no change of TWEAK protein levels in the substantia nigra or the striatum of human PD patients as compared to matched control subjects. Mitigating the effects of endogenous TWEAK protein using neutralizing antibody did affect MPTP-mediated neurotoxicity in the substantia nigra using the sub-acute model of MPTP (30mg/kg i.p. over five consecutive days). Neither TWEAK nor Fn14 genetic ablation led to attenuation of MPTP-toxicity in the acute model. These findings suggest that TWEAK signaling might be an aspect of MPTP-mediated neuropathology and be involved in the overall neurodegenerative pathology of PD., (Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.)

In this study, different extraction procedures based on the QuEChERS method were compared for the multiresidue determination of pesticides in orange juice by ultra high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). After choosing preliminary conditions, an experimental design was carried out with the variables C18, PSA, NaOH and CH3COONa to optimize the sample preparation step. The validation results of the validation were satisfactory, since the method presented recoveries between 70% and 118%, with RSD lower than 19% for spike levels between 10 and 100 μg L(-1). The method limit of detection (LOD) and limit of quantification (LOQ) ranged from 3.0 to 7.6 μg L(-1) and from 4.9 to 26 μg L(-1), respectively. The method developed was adequate for the determination of 74 pesticide residues in orange juice., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

B19V infection is common during childhood. It is self-limited in healthy individuals, but is often associated with transient aplastic crisis in children with sickle cell disease. The aim of this study was to estimate the prevalence and incidence of B19V infection in children with sickle cell disease screened by the Newborn Screening Program of Minas Gerais, Brazil, and followed-up at Fundação Hemominas. Serum or plasma samples from 278 patients were tested for anti-B19V IgG and IgM using commercial ELISA and for viral DNA using in-house real-time PCR assays; 127 negative-children were retested about 1 year later. The median age of children at first testing was 5.9 years (0.8-12.3). The estimated prevalence of B19V was 29.5 % (95%CI 24.1-34.9 %). The incidence of B19V in those 127 negative-children was 18.2 cases/100 patient-years. All DNA-positive samples were identified as genotype 1, except one sample, in which both genotypes 1 and 3 were identified. It was observed that the higher the child's age, the higher the probability of B19V infection. The analysis of clinical and hematological data showed a significant association of B19V infection with transient aplastic crisis and acute splenic sequestration, higher frequency of transfusions, and higher rate of hospitalization, but not with acute chest syndrome or stroke. These results emphasize the impact of B19V infection on the course of sickle cell disease. Strategies to prevent and monitor B19V infection in children with sickle cell disease should be considered to diminish its morbidity in this susceptible population., (© 2015 Wiley Periodicals, Inc.)

The most common cancer treatments currently available are radio- and chemo-therapy. These therapies have, however, drawbacks, such as, the reduction in quality of life and the low efficiency of radiotherapy in cases of multiple metastases. To lessen these effects, we have encapsulated an anti-cancer drug into a biocompatible matrix. In-vitro assays indicate that this bio-nanocomposite is able to interact and cause morphological changes in cancer cells. Meanwhile, no alterations were observed in monocytes and fibroblasts, indicating that this system might carry the drug in living organisms with reduced clearance rate and toxicity. X-rays and neutrons were used to investigate the carrier structure, as well as to assess the drug mobility within the bio-nanocomposite. From these unique data we show that partial mobility restriction of active groups of the drug molecule suggests why this carrier design is potentially safer to healthy cells.

The aim of this work was to investigate the kinetics of removal of clomazone herbicide from an aqueous solution by electrocoagulation. The experiments were performed in a cylindrical batch reactor with six aluminum electrodes in monopolar mode, arranged in series and connected to a digital DC power. The aqueous solution (tap water + clomazone) with initial pH close to 7.9 was always treated at ambient temperature (≈20 °C) and atmospheric pressure for 5,400 s. For a confidence level of 95% the rate constant of electrocoagulation and the efficiency of removal of clomazone at equilibrium were 2.1 × 10(-3) ± 0.5 × 10(-3) s(-1) and 97.7 ± 2.2%, respectively. The final chemical oxygen demand was 88% lower than that measured initially, while turbidity and apparent color were totally removed from the synthetic solution at a rate close to that of formation of aluminum hydroxides. Some reaction intermediates, such as benzonitrile-2-chloro and 2-chloro-hex-2,4-diene-1,6-dioic-acid determined by gas chromatography mass spectrometry (GC-MS) analysis, explain the ratio of equilibrium to initial total organic carbon approximately between 0.6 and 0.8 at a probability of 95%.

The process of cooling and cryopreservation of prawn embryos is a viable alternative for a continuous supply of larvae for freshwater prawn farming ponds. However, studies involving the application of those techniques as well as on toxicity of cryoprotectants in freshwater prawn embryos are scarce. Thus, this study aims to test the toxicity of methylic alcohol (MET), dimethyl sulfoxide (DMSO) and ethylene glycol (EG) on Macrobrachium amazonicum embryos. For the present experiment, pools of embryos were taken from 15 M. amazonicum females and were divided into three groups and tested in duplicate at concentrations of 10, 5, 3; 1, 0.5 or 0.1%. Toxicity tests were conducted for 24 h in Falcon® pipes to obtain the lethal concentration for 50% of the larvae (LC50). After the set period for testing, random samples of embryos were removed for morphological analysis under stereoscopic microscopes. Results were analysed using analysis of variance (ANOVA) and Tukey's test at a 5% significance level and Trimmed Spearman-Karber Analysis to determine LC50-24 h. DMSO toxicity tests revealed that 5% and 10% concentrations showed the highest toxicity and differed from the control (P ≤ 0.05), 24h-LC50 was 437.4 ± 14.4 µL. MET was less toxic among the tested cryoprotectants and concentrations did not allow the determination of its LC50-24h. For tests with EG, concentrations of 3, 5 or 10% solutions resulted in a 100% mortality to tested embryos; EG was the tested cryoprotectant with the highest toxicity, with an LC50-24h average of 81.91 ± 35.3 µl.

Several environmental parameters may influence biological processes of several aquatic invertebrates, such as the Monogenea. Current analysis investigates oviposition, hatching success and infestation of Aphanoblastella mastigatus, a parasite of the silver catfish Rhamdia quelen at different temperatures (~ 24 and 28 °C) and salinity (by adding sodium chloride to water, at concentrations 0, 5 and 9 g/L) in laboratory. There was no significant difference in oviposition rate and in A. mastigatus infestation success at 24 and 28 °C. On the other hand, the concentration 9 g/L of sodium chloride in the water impaired the parasite's survival and the viability of the eggs. Results show that its usage is efficient as a possible prophylactic treatment. Eclosion rate of A. mastigatus's eggs was significantly higher at 28 °C, although it was significantly less from 5 g/L. Two oviposition peaks (06h15 and 18h15) occurred during a 24-hour period, or rather, during the highest variations in luminosity. Further studies are recommended with greater temperature intervals and more intense experimental infestations to verify the effects of temperature in the life span and infestation success of A. mastigatus.

Third-stage larvae (L3) of Hysterothylacium sp. were collected by the first time in juveniles of pirarucu Arapaima gigas farmed in the Rio Preto da Eva, Amazonas state. Ninety-eight (98) out of 100 examined fish showed to be parasitized. Five hundred and ninety larvae of Hysterothylacium sp. were collected from the intestines, stomach and pyloric caeca. The mean intensity of parasite indexes was 6.02 (±5.75) ranging from 1 to 40 larvae per host and the mean abundance was 5.9 (±5.76). The A. gigas is the new host record for larvae of Hysterothylacium sp. in Brazil, and this is the first record of larvae of Hysterothylacium (Nematoda: Anisakidae) with zoonotic potential in the pirarucu from South America.

Cryptococcosis caused by the fungus Cryptococcus neoformans is an opportunistic mycosis, infecting mainly immunodepressed individuals. Molecular epidemiology studies of cryptococcosis in Europe are limited. This paper presents a retrospective study of cryptococcosis in 105 cryptococcal isolates from two hospitals in Lisbon, Portugal, among HIV/AIDS patients, from 1991 to 2007. Among these patients, the number of cases of cryptococcosis increased from 5.1 to 6.9 cases per year from the pre- to post-highly active antiretroviral therapy (HAART) era. As expected, the median age of the patients increased, from 32 (mean: 33 ± 8) to 39 (mean: 41 ± 10) years, and the ratio of male to female patients remained high (7.7 and 7.6, respectively). Strain genotyping based on restriction fragment length polymorphism of the orotidine monophosphate pyrophosphorylase (URA5-RFLP) gene showed that, in general, the relative frequencies of the genotypes VNI-IV are similar to those from other European countries. These frequencies were, respectively, for the pre- and post-HAART periods: 41.7 and 43.5 % for VNI; 2.8 and 17.4 % for VNII; 38.9 and 30.4 % for VNIII; 16.7 and 7.2 % for VNIV and 0 and 1.4 % for VGII. Some apparent although statistically insignificant differences among these values were observed between both periods. The genotypic frequencies were not also statistically different according to the patients' gender or age range. Of note are the high proportion of VNIII isolates (common in Europe) and the high increase in the frequency of the VNII genotype in the post-HAART. Ultimately, these results may have implications in disease therapy, management and control.

Objectives: In this study, we present a strategy for RHD gene screening based on real-time polymerase chain reaction (PCR) using dried blood spots of pooled samples., Background: Molecular analysis of blood donors may be used to detect RHD variants among the presumed D-negative individuals. RHD genotyping using pooled samples is a strategy to test a large number of samples at a more reasonable cost., Materials and Methods: RHD gene detection based on real-time PCR using dried blood spots of pooled samples was standardised and used to evaluate 1550 Brazilian blood donors phenotyped as RhD-negative. Positive results were re-evaluated by retesting single samples using real-time PCR and conventional multiplex PCR to amplify five RHD-specific exons. PCR-sequence-specific primers was used to amplify RHDψ allele., Results: We devised a strategy for RHD gene screening using dried blood spots of five pooled samples. Among 1550 serologically D-negative blood donors, 58 (3.74%) had the RHD gene. The non-functional RHDψ allele was detected in 47 samples (3.02%)., Conclusion: The present method is a promising strategy to detect the RHD gene among presumed RhD-negative blood donors, particularly for populations with African ancestry., (© 2015 British Blood Transfusion Society.)

Background: The RHD gene is highly polymorphic, which results in a large number of RhD variant phenotypes. Discrepancies in RhD typing are still a problem in blood banks and increase the risk of alloimmunization. In this study, the RhD typing strategy at a blood bank in Brazil was evaluated., Methods: One-hundred and fifty-two samples typed as RhD negative and C or E positive by routine tests (automated system and indirect antiglobulin test using the tube technique) were reevaluated for RhD status by three methods. The method with the best performance was implemented and evaluated for a period of one year (n=4897 samples). Samples that were D positive exclusively in the confirmatory test were submitted to molecular analysis., Results: The gel test for indirect antiglobulin testing with anti-D immunoglobulin G (clone ESD1) presented the best results. Seventy samples (1.43%) previously typed as RhD negative showed reactivity in the gel test for indirect antiglobulin testing and were reclassified as D positive. D variants that may cause alloimmunization, such as weak D type 2 and partial D(VI), were detected., Conclusion: The confirmatory RhD test using the gel test for indirect antiglobulin testing represents a breakthrough in transfusion safety in this blood center. Our results emphasize the importance of assessing the blood group typing strategy in blood banks., (Copyright © 2015 Associação Brasileira de Hematologia, Hemoterapia e Terapia Celular. Published by Elsevier Editora Ltda. All rights reserved.)

This study investigated the immunization by intraperitoneal injection (i.p.) (assay I) and immersion bath (assay II) with live theronts of Ichthyophthirius multifillis in Rhamdia quelen and its influence on the hemato-immunological and biochemical parameters. Fish were divided in control (non immunized no challenged); non immunized and challenged with 12,000 theronts/fish; non immunized and challenged with 22,000 theronts/fish; immunized and challenged with 12,000 theronts/fish; immunized and challenged with 22,000 theronts/fish. Six days after challenge, either in the assay I or in the assay II the prevalence of I. multifillis in the gills was higher in non immunized fish (33.33% and 27.77%, respectively). In the assay I showed higher numbers of thrombocytes, leukocytes, lymphocytes, neutrophils and monocytes 20 days after injection and lower numbers after challenge. The immunoglobulin values were higher in fish non immunized. Fish immunized by immersion bath (assay II) showed greater values of catalase (CAT) in the liver (1245.49 U/mgprt) when compared to i.p. (198.79 U/mgprt). The levels of CAT in the liver of fish from the assay II were greater (1738.47 U/mgprt) 14 days after immunization than that observed 21 days after (1114.26 U/mgprt). The vaccination by i.p method showed influence on the hematological parameters. On the other hand, the immersion bath vaccination showed greater influence on the catalase activity in the liver. The results showed that new parameters like total protein, immunoglobulin and antioxidant enzymes could be considered in evaluating the host response to infection., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

This study evaluated the correlation of hematological parameters with the mean abundance of the monogenean helminth Rhabdosynochus rhabdosynochus in Centropomus undecimalis reared at different temperatures and salinities. The experimental conditions were: 28 °C/0 ppt (parts per thousand); 28 °C/15 ppt; 28 °C/32 ppt; 25 °C/0 ppt; 25 °C/15 ppt; and 25 °C/32 ppt. The prevalence was 100.0% in fish at 28 °C/15 ppt, 28 °C/32 ppt and 25 °C/15 ppt, which was significantly different (p < 0.05) from those at 25 °C/32 ppt. The red blood cell (RBC) count, hematocrit and total leukocyte (WBC) count were significantly higher in fish at 28 °C/15 ppt and 28 °C/32 ppt. The mean abundance of R. rhabdosynochus, hematocrit and RBC showed positive correlations (P < 0.05) with temperature (ρ= 0.3908; ρ= 0.4771 and ρ = 0.2812). Mean abundance showed negative correlations with hemoglobin (ρ= -0.3567) and mean corpuscular hemoglobin concentration (MCHC) (ρ = -0.2684). No correlation between abundance and salinity was detected among the experimental conditions (ρ = -0.0204). The low numbers of monogeneans recorded (min -1 and max -33) explain the few changes to fish health. This suggests that these experimental conditions may be recommended for development of rearing of C. undecimalis in Brazil, without any influence or economic losses from R. rhabdosynochus.

Cryopreservation has not been used successfully to preserve fish embryos, although chilling techniques have been used with good results. The aim of this study was to chill Piaractus brachypomus embryos at - 10°C for various storage times. Embryos at the following ontogenetic stages were used: blastoderm - 1.2 hours post-fertilization (hpf); epiboly - 5 hpf; blastopore closure - 8 hpf; and appearance of the optic vesicle - 13 hpf. One hundred embryos were selected from each ontogenetic stage and chilled at - 10°C for 6 or 10 h. The results were analysed using analysis of variance (ANOVA) and Tukey's test at a 5% significance level. A significantly greater number of completely developed live larvae were observed following embryonic treatment with a cryoprotectant solution that contained 17.5% sucrose and 10% methanol. There was no survival for embryos cooled at - 10°C in initial developmental stages (1, 2 and 5 h hpf). Furthermore, higher survival rates were observed when embryos were treated at more advanced developmental stages (8 and 13 hpf). Therefore, P. brachypomus embryos at the blastopore-closure (8 hpf) or appearance-of-optic-vesicle (13 hpf) stages should be used for embryo chilling protocols and chilling should be performed using a 17.5% sucrose with a 10% methanol solution at - 10°C for up to 6 h. The best results were obtained with 13-hpf and 8-hpf embryos and cooling at 6 h of storage.

Although there are several reports of Myxobolus species infecting the somatic muscle of teleost fish, species of this genus have not been described parasitizing the muscle tissue of pacus, Piaractus mesopotamicus. This study presented the first report of natural infection by Myxobolus sp. in the skeletal muscle of pacus reared in intensive system. Twenty-one fish (±142.2 g; ±23.1 cm) were captured (April 2013) from an intensive fish farm in São Paulo State, southeast Brazil. Spores of Myxobolus sp. were contained within plasmodia and showed oval morphology with the apical portion slightly pyriform and two polar capsules pyriform retaining the same ratio to each other. In the histopathological study, the skeletal muscle do not present signs of inflammation. The integrity of myofibrils within the infected fibers showed some degree of degeneration, with partial loss of myofibrillar details and striations. Spores were found infecting the skeletal muscle of 18 fish (85.7%). Finally, the high prevalence of Myxobolus sp. infection in the skeletal muscle of P. mesopotamicus and the absence of macroscopic lesions in the muscle tissue indicate the necessity of more meticulous examinations during the health management in the rearing system and at slaughter of pacus to ensure best quality of meat.

Objective: It has been suggested that a greater dairy consumption is a contributing factor to a lower prevalence of metabolic syndrome (MetS) as a result of the possible actions of some milk nutrients. However, results remain controversial. The aim of this study was to examine the association between dairy consumption and MetS and its components., Methods: Dairy consumption and biochemical and anthropometric parameters were determined in 2031 young adults ages 23 to 25 y. Food consumption was assessed using a food frequency questionnaire. The estimated habitual portion of dairy products consumed daily was divided into quintiles. The criteria of the International Diabetes Federation (IDF) and of the Joint Interim Statement (JIS) were used for the classification of MetS. Nonadjusted and adjusted odds ratios (ORs) were estimated by logistic regression., Results: The prevalence of MetS was 11.9% by the IDF criteria and 9% by the JIS criteria. A greater dairy consumption was associated with a lower prevalence of MetS according to both IDF (OR, 0.61; 95% confidence interval [CI], 0.38-0.97) and JIS (OR, 0.62; 95% CI, 0.39-0.98) criteria when the last quintile was compared with the first. The association persisted in the model adjusted for demographic, socioeconomic, dietary, and lifestyle variables according to the IDF (OR, 0.53; 95% CI, 0.30-0.93) and was borderline according to the JIS (OR, 0.59; 95% CI, 0.34-1.00), but lost significance when data were adjusted for calcium in both models., Conclusions: A greater dairy consumption is associated with a lower prevalence of MetS, with calcium probably being the nutrient responsible for this association., (Copyright © 2015 Elsevier Inc. All rights reserved.)

In this study, an important ornamental fish, Betta splendens (Osphronemidae), from three different Brazilian states was examined for parasitic infestations. Smears with parasites were impregnated with silver nitrate or stained using Giemsa for taxonomic evaluation. A disc-shaped trichodinid with a body diameter of 39.7 ± 3.3 µm, adhesive disc diameter of 32.9 ± 3.1 µm and denticulate ring diameter of 19.5 ± 2.0 µm was found. The morphological characteristics resembled those of Trichodina modesta Lom, 1970, a species that shows clear host specificity for Cypriniformes. Until now, its occurrence was restricted to the Eurasian region. In the present study, a new host for T. modesta is reported and therefore the first occurrence of this species in the Americas. The parasite was possibly introduced into the Neotropical region through the exotic fish trade, especially of Cypriniformes used by aquarists. The distribution of this ciliate is discussed and a checklist of localities and hosts for the species is provided.

The practice of refrigerating raw milk at the farm has provided a selective advantage for psychrotrophic bacteria that produce heat-stable proteases and lipases causing severe quality problems to the dairy industry. In this work, a protease (AprX) and a lipase (LipM) produced by Pseudomonas fluorescens 041, a highly proteolytic and lipolytic strain isolated from raw milk obtained from a Brazilian farm, have been purified and characterized. Both enzymes were purified as recombinant proteins from Escherichia coli . The AprX metalloprotease exhibited activity in a broad temperature range, including refrigeration, with a maximum activity at 37 °C. It was active in a pH range of 4.0 to 9.0. This protease had maximum activity with the substrates casein and gelatin in the presence of Ca (+2) . The LipM lipase had a maximum activity at 25 °C and a broad pH optimum ranging from 7.0 to 10. It exhibited the highest activity, in the presence of Ca (+2) , on substrates with long-chain fatty acid residues. These results confirm the spoilage potential of strain 041 in milk due to, at least in part, these two enzymes. The work highlights the importance of studies of this kind with strains isolated in Brazil, which has a recent history on the implementation of the cold chain at the dairy farm.

Alternate Current Biosusceptometry (ACB) is a promising bio-magnetic method, radiation free and easily performed used for gastric emptying exams. Due to development on its sensitivity level, interesting nature, noninvasiveness and low cost it has attracted a lot of attention. In this work, magnetic nanoparticles of Mn-Zn ferrite as well as dextrose-modified nanoparticles were synthesized to be used as possible tracers in ACB gastric emptying exams. In addition, a magnetic muco-adhesive gel was obtained by modifying the ferrite nanoparticles with cellulose. Based on in-vivo tests in rats, we show that the pure ferrite nanoparticles, whose isoelectric point was found to be at pH=3.2, present a great sensitivity to pH variations along the gastrointestinal tract, while the reduction of the isoelectric point by the dextrose modification leads to suitable nanoparticles for rapid gastric emptying examinations. On the other hand, the in-vivo tests show that the muco-adhesive cellulose gel presents substantial stomach adhesion and is a potential drug delivery system easily traceable by the ACB system., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The Phylum Protozoa brings together several organisms evolutionarily different that may act as ecto or endoparasites of fishes over the world being responsible for diseases, which, in turn, may lead to economical and social impacts in different countries. Apart from the recent advances for the diagnosis of fish diseases in Brazil, little is known on the protozoan parasites and their relationship with environment and host. This revision presents the most important protozoan parasites found in farmed fish from Brazil, not only with emphasis on its diagnosis, biology, transmission and host-parasite relationship, but also on some information that may be useful to researchers in determining the correct diagnosis in fish farms.

This study evaluated the influence of diet supplementation with propolis and Aloe barbadensis on hematological parameters and parasitism in tilapia. One hundred and eighty fish were distributed among 12 water tanks forming four treatments: fish supplemented with a 1:1 mixture of 0%, 0.5%, 1% and 2% propolis and aloe extracts. After the fish had been fed on the experimental diets for 15 and 21 days, blood samples were taken and parasites collected. The monogeneans Cichlidogyrus sclerosus, C. halli, C. thurstonae and Scutogyrus longicornis were identified in the gills. Between the sampling times, there were increases in the numbers of erythrocytes, leukocytes, thrombocytes and lymphocytes, as observed after 21 days, possibly due to the stress level over the course of the assay and/or accumulation of substances in the organism. Supplementation with the mixture of propolis and aloe for 15 days showed the highest efficacy against the parasites. This was possibly due to the association between the two compounds. The results demonstrated that supplementation with mixtures of extracts did not produce hematological alterations and also favored a significant reduction in the number of gill parasites. The best results were achieved after 15 days of feeding with a diet with 0.5% and 1% supplementation with the extract mixture, which increased efficiency by 83 and 85% respectively.

Cucullanus tucunarensis n. sp. (Cucullanidae) is described parasitizing the peacock bass Cichla piquiti (Cichlidae) from the Tocantins River, Tocantins State, Brazil. The new species is unique and differs from all its congeners by having a tail tip provided with several sclerotized spine-like processes on its ventral side, present in both the male and female. Furthermore, C. tucunarensis n. sp. is compared with other species of the genus recorded in the same zoogeographical region and in the same group of hosts (Perciformes).

Background: Human T-lymphotropic virus 1 (HTLV-1) has been associated with leukemia/lymphoma (ATL) and myelopathy/tropical spastic paraparesis (HAM/TSP), in addition to other inflammatory diseases as well as infection complications. Therapeutic approaches for HTLV-1-related pathologies are limited. The labdane diterpene myriadenolide (AMY) is a natural product that exhibit biological activities, such as anti-inflammatory and antiviral activity as reported for HIV and herpesvirus., Results: We demonstrated that this natural product was able to inhibit the expression of gag-pol mRNA and substantially reduced the expression of the structural proteins p19 and gp46. Comparison of treated and untreated cells shows that AMY alters both the morphology and the release of viral particles. The Atomic Force Microscopy assay showed that the AMY treatment reduced the number of particles on the cell surface by 47%., Conclusion: We demonstrated that the labdane diterpene myriadenolide reduced the expression of the structural proteins and the budding of viral particles, besides induces altered morphogenesis of HTLV-1, conferring on AMY a new antiviral activity that may be useful for the development of new compounds with specific anti-HTLV-1 activity.

Numerous bacteria coordinate gene expression in response to small signalling molecules in many cases known as acylhomoserine lactones (AHLs), which accumulate as a function of cell density in a process known as quorum sensing. This work aimed to determine if phenotypes that are important to define microbial activity in foods such as biofilm formation, swarming motility and proteolytic activity of two Pseudomonas fluorescens strains, isolated from refrigerated raw milk, are influenced by AHL molecules. The tested P. fluorescens strains did not produce AHL molecules in none of the evaluated media. We found that biofilm formation was dependent on the culture media, but it was not influenced by AHLs. Our results indicate that biofilm formation, swarming motility and proteolytic activity of the tested P. fluorescens strains are not regulated by acyl-homoserine lactones. It is likely that AHL-dependent quorum sensing system is absent from these strains.

In multicellular organisms, cell motility is central in all morphogenetic processes, tissue maintenance, wound healing and immune surveillance. Hence, failures in its regulation potentiates numerous diseases. Here, cell migration assays on plastic 2D surfaces were performed using normal (Melan A) and tumoral (B16F10) murine melanocytes in random motility conditions. The trajectories of the centroids of the cell perimeters were tracked through time-lapse microscopy. The statistics of these trajectories was analyzed by building velocity and turn angle distributions, as well as velocity autocorrelations and the scaling of mean-squared displacements. We find that these cells exhibit a crossover from a normal to a super-diffusive motion without angular persistence at long time scales. Moreover, these melanocytes move with non-Gaussian velocity distributions. This major finding indicates that amongst those animal cells supposedly migrating through Lévy walks, some of them can instead perform q-Gaussian walks. Furthermore, our results reveal that B16F10 cells infected by mycoplasmas exhibit essentially the same diffusivity than their healthy counterparts. Finally, a q-Gaussian random walk model was proposed to account for these melanocytic migratory traits. Simulations based on this model correctly describe the crossover to super-diffusivity in the cell migration tracks.

Although human T-cell lymphotropic viruses (HTLV-1/2) were described over 30 years ago, they are relatively unknown to the public and even to healthcare personnel. Although HTLV-1 is associated with severe illnesses, these occur in only approximately 10% of infected individuals, which may explain the lack of public knowledge about them. However, cohort studies are showing that a myriad of other disease manifestations may trouble infected individuals and cause higher expenditures with healthcare. Testing donated blood for HTLV-1/2 started soon after reliable tests were developed, but unfortunately testing is not available for women during prenatal care. Vertical transmission can occur before or after birth of the child. Before birth, it occurs transplacentally or by transfer of virus during cesarean delivery, but these routes of infection are rare. After childbirth, viral transmission occurs during breastfeeding and increases with longer breastfeeding and high maternal proviral load. Unlike the human immunodeficiency virus types 1 and 2, HTLV is transmitted primarily through breastfeeding and not transplacentally or during delivery. In this study, we review what is currently known about HTLV maternal transmission, its prevention, and the gaps still present in the understanding of this process., (© The Author 2014. Published by Oxford University Press on behalf of the Pediatric Infectious Diseases Society. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.)

Although there is evidence on the benefits in the use of immunostimulants in aquaculture, there are few commercial products being used. This study evaluated the use of natural substances as potential sources for the production of immunostimulants. Propolis and Aloe barbadensis have been widely studied and its extracts have different chemical constituents responsible for antimicrobial, anti-inflammatory and immunostimulant. Tilapia juveniles were fed for two weeks with diets supplemented mix of propolis extracts and aloe (1:1) in different concentrations: 0.5, 1 e 2%. After the experimental period, fish blood was collected for hematoimmunological as follows : hematocrit, total plasma protein, erythrocytes (RBC), leukocytes (WBC), differential leukocyte count, phagocytic activity, serum lysozyme activity, and serum antimicrobial activity, serum antimicrobial activity (evaluated against Aeromonas hydrophila, Enterococcus durans and Escherichia coli). Except for higher number of thrombocytes in 1%-supplemented fish, the rest did not show significant difference., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

A modified version of the QuEChERS method has been evaluated for the determination of 21 pharmaceuticals and 6 personal care products (PPCPs) in drinking-water sludge samples by employing ultra high liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The performance of the method was evaluated through linearity, recovery, precision (intra-day), method detection and quantification limits (MDL and MQL) and matrix effect. The calibration curves prepared in acetonitrile and in the matrix extract showed a correlation coefficient ranging from 0.98 to 0.99. MQLs values were on the ng g(-1) order of magnitude for most compounds. Recoveries between 50% and 93% were reached with RSDs lower than 10% for most compounds. Matrix effect was almost absent with values lower than 16% for 93% of the compounds. By coupling a quick and simple extraction called QuEChERS with the UPLC-MS/MS analysis, a method that is both selective and sensitive was obtained. This methodology was successfully applied to real samples and caffeine and benzophenone-3 were detected in ng g(-1) levels., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Pollution of water sources by pesticides is a result of intensive use of these compounds, and the establishment of analytical methods for monitoring programs is very important. This paper presents a method for the determination of multiclass pesticide residues in rice paddy water based on SPE and GC/MS/MS. The validation results were satisfactory. All compounds showed adequate linearity (r2 >or= 0.99), and analysis by GC/MS/MS with a triple quadrupole mass spectrometer gave high selectivity and sensitivity. With the proposed SPE step an LOD of 0.06 microg/L was achieved. Recoveries from blank samples spiked at 0.2, 04, and 0.8 microg/L were between 70 and 120% with RSD <20% for most analyzed compounds, despite their different chemical nature, indicating good accuracy and precision. The proposed method is efficient for pesticide residue determination, including some metabolites, and gave good performance when applied to real samples.

The aim of the present study was to evaluate the serological methods using ELISA with recombinant-rK39 (ELISA-rK-39) and soluble extract-SE (ELISA-SE) antigens, the indirect fluorescence antibody test (IFAT) in comparison to an immunochromatography rapid diagnostic test (RDT-rK39) and with a direct parasitological exam (PA) for Canine Visceral Leishmaniasis (CVL) diagnosis. The results showed that 89% (60/67) of the dogs were positive for at least one serological diagnostic test. ELISA-SE was the test that detected anti-Leishmania antibodies in the serum of the highest number of dogs (71.6%) followed by ELISA-rK39 (65.7%), IFAT (65.7%) and RDT-rK39 (55.2%). PA detected the lowest numbers (40.3%) of positive dogs. In relation to the total of examined dogs, the Kappa indexes (p ≤ 0.05) showed a good agreement between ELISA-SE and IFAT (88.1%; k = 0.7237), and it was also observed in the comparison of RDT-rK39 with ELISA-SE (83.6%, k= 0.6561), IFAT (83.5%, k= 0.6605) and PA (85.0%, k= 0.7074). A bad agreement was detected in any association of ELISA-rk39 with the other tests in either symptomatic or asymptomatic animals. ELISA as well as RDT using recombinant antigenic protein (rK39) were the methods that detected the lowest prevalence rates (33.3%) of CVL in asymptomatic dogs. In conclusion, only one test does not adequately identify dogs with CVL and it is necessary the association of two or more diagnostic tests. Because of the good agreement indexes of RDT-rK39 when evaluated with ELISA-SE, IFAT and PA it was suggested as a complementary method to be used in association with either ELISA-SE or IFAT, particularly in the symptomatic dogs. Furthermore, new studies are recommended in order to improve the sensitivity of tests mainly for asymptomatic dogs.

Prochilodus lineatus is a freshwater fish species found in South America. It is common in aquaculture, but few studies regarding diseases of this fish have been performed. This study presents data of the occurrence of Trichodina heterodentata Duncan, 1977, as well as the pathological alterations detected by light and scanning electron microscopy (SEM). Twenty 20-day-old larvae were harvested from an earth pond and examined. Larvae showed erratic swimming on the pond edges and some had a whitish tegument. Larval smears were either impregnated with silver nitrate or stained with Giemsa stain to observe the taxonomic features of the ciliates. Five larvae were fixed in formalin solution for histopathological analysis, and another five specimens were fixed in glutaraldehyde for SEM. All larvae were diagnosed with a severe infestation by trichodinid T. heterodentata. Histological sections showed discrete hyperplasia of the gill filaments with subepithelial oedema of the secondary lamellae. In the SEM, suction areas were observed on the skin, gill and eye; corrosion and ulceration of the fins were associated with the bacterial presence of cocci on the lesions. This is the first report of T. heterodentata in P. lineatus that is responsible for an acute disease that culminates in larval mortality.

This study evaluated for two years the haematological parameters and the occurrence of gill parasites in silver catfish Rhamdia quelen, farmed in the state of Santa Catarina, southern Brazil. Between March 2010 and January 2012, a total of 137 fish were caught in a fish farm to compare the haematological and parasitological analysis, during hot and cold seasons. Simultaneously, water quality parameters were measured in the pond to analyze the relationship between environmental conditions and haematology and parasitism. During the hot season, there was an increase in water temperature, electrical conductivity and ammonia followed by a decrease in dissolved oxygen, pH, transparency and alkalinity. Two species of Monogenea were observed in the gills, Aphanoblastella mastigatus (Dactylogyridae) and Scleroductus sp. (Gyrodactylidae). Additionally, during this season, there was a significant increase (p<0.05) in the mean intensity of infection by Monogenea, as well as total leukocytes number, thrombocytes and lymphocytes, and a reduction in monocytes. The changes might be caused by parasitism and/or environmental variations between seasons.