Efeito de um extrato de Ganoderma lucidum (Reishi) na marcação de constituintes sanguíneos com tecnécio-99M e na sobrevivência de Escherichia coli

Clinical evaluations have been made possible with radiobiocomplexes marked with tecnecium-99m (99mTc). Natural or synthetic drugs are able to interfere in the marking of blood structures with 99m Tc. Also, the toxicity of several natural products has been described. The aim of this study was evaluating the effect of an extract of Ganoderma lucidum (Reishi) in the marking of blood constituents with 98m Tc and in the survival of Escherichia coli. Blood samples from Wistar rats were treated with reishi extract. Radiomarking procedure was performed. Samples of plasma (P), blood cells (CS), and insoluble (FI) and soluble (FS) fractions of P and CS were separated and the radioactivity was counted to determine radioactivity percentages (%ATI). Escherichia coli AB1157 cultures were treated with stannous chloride in the presence and absence of the reishi extract. Blood samples and bacterial cultures treated with NaCl 0.9% were used as controls. Data indicated that the reishi extract has significantly altered (p<0,05) the %ATI of P, CS, FI-P, FS-P, FI-CS e FS-CS, as well as it has increased survival of bacterial cultures treated with stannous chloride. Our results suggest that the Reishi extract would be able to present a redox/ chelant action by altering blood constituent marking with 99mTc and by protecting bacterial cultures against stannous chloride-induced oxydating lesions. The study had a multidisciplinary character, with the participation of the following areas of knowledge: Biophysics, Radiobiology, Botanics, Phytotherapy, and Hematology

Avaliação de efeitos biológicos de antiinflamatórios derivados do ácido propiônico através de modelos experimentais em nível molecular e celular

Derivatives of propionic acid NSAIDs are irreversible inhibitors of cyclooxygenase enzyme widely used. The aim of this study was to evaluate, through different experimental models, biological effects of derivatives of propionic acid (fenoprofen, naproxen, ibuprofen and ketoprofen) in cellular and molecular level. The labeling of blood constituents with technetium-99m (99mTc) and morphological analysis of erythrocytes of blood of rats, as well as growth, survival of cultures of Escherichia coli (E. coli) and the assessment of bacterial plasmid electrophoretic profiles were models used for experimental evaluation of possible biological effects of antiinflammatory drugs. The results show that, in general, anti-inflammatory drugs evaluated were not able to alter the labeling of blood constituents with 99mTc, the morphology of red blood cells from blood of rats, as well as the growth of cultures of E. coli and the electrophoretic profile of plasmid DNA. However, naproxen appears to cytotoxic effect on bacterial cultures, plasmids and genotoxic effects in reducing the action of stannous chloride in cultures of E. coli. The use of experimental fast performance and low cost was important for assessment of biological effects, contributing to a better understanding of the properties of propionic acid derivatives studied. anti-inflammatory, blood constituents, technetium-99m, stannous chloride, Escherichia coli; DNA

Hematology and productive performance of nile tilapia (Oreochromis niloticus) naturally infected with Flavobacterium columnare

Columnaris disease is one of the main causes of mortality in tilapia rearing and is responsible for large economic losses worldwide. Hematology is a tool that makes it possible to study organisms' physiological responses to pathogens. It may assist in making diagnoses and prognoses on diseases in fish populations. The hematological variables of nile tilapia were studied in specimens with a clinical diagnosis of columnaris disease and in specimens that were disease-free. The total erythrocyte count, hemoglobin rate, hematocrit percentage, mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), mean corpuscular volume (MCV), organic defense blood cell percentages (leukocytes and thrombocytes) and hepatosomatic and splenosomatic index were determined. The results showed that there were changes in the erythrocytic series and in organic defense blood cells, in the fish infected with the bacterium, with reductions in erythrocytic variables and significant increases in the numbers of circulating lymphocytes and neutrophils.

Análise in vitro da atividade antifúngica e de toxicidade da anfotericina B pré-aquecida

The aim of this work was to evaluate how an aqueous micellar system containing Amphotericin B (AmB) and sodium deoxycholate (DOC) can be rebuilt after heating treatment. Also a review of the literature about the new physicochemical and biological properties of this new system was carried out. Afterwards, heated (AmB-DOC-H) and unheated (AmB-DOC) micelles were subsequently diluted at four different concentrations (50mg.L-1, 5mg.L-1, 0.5mg.L-1 and 0.05mg.L-1) to perform the physicochemical study and, then, the pharmacotoxicity assay, in which two cell models were used for the in vitro experiments, Red Blood Cells (RBC) from human donors and Candida parapisilosis (Cp). While potassium (K+) and hemoglobin leakage from RBC were the used parameters to evaluate the acute and chronic toxicity, respectively, the efficacy of AmB-DOC and AmB-DOC-H were assessed by K+ leakage and cell survival rate from Cp. The spectral study revealed a slight change on the aggregate peak from 327nm to 323nm for AmB-DOC-H compared to AmB-DOC. Concerning the toxicity, although AmB-DOC and AmB-DOC-H presented different behavior for hemoglobin leakage, AmB-DOC produced higher leakage than AmB-DOC-H at high concentrations (from 5mg.L-1) with values tending to zero. However, concerning K+ leakage, both AmB-DOC and AmB-DOC-H, showed similar profile for both cell models, RBC and Cp (p<0,05). AmB-DOC-H and AmB-DOC also revealed similar profile of activity against Cp with equivalent survival rate. In short, the AmB-DOC-H showed much less toxicity than AmB-DOC, but remained as active as the late one against fungal cell. Therefore, the results highlight the importance of this new procedure as a simple, inexpensive and safe alternative to produce a new kind of micelle system for treatment of systemic fungal infections

Biological indicators of stress in pacu (Piaractus mesopotamicus) after capture

Os efeitos da captura (perseguição, contenção em puçá e exposição aérea) no perfil sanguíneo do cortisol, glicose, cloreto, sódio, potássio, cálcio e na osmolaridade, hematócrito, hemoglobina, número de células vermelhas (CV) e volume corpuscular médio (VCM) foram investigados no pacu (Piaractus mesopotamicus). Um total de 132 peixes (49,7 ± 11,7 g) foi submetido à captura com 3 ou 5 minutos de exposição aérea. Nove peixes de cada tratamento foram amostrados 5, 15, 30, 60 minutos e 24 horas depois e outros nove peixes foram amostrados antes da captura e considerados controle. A captura resultou em aumento do cortisol e glicose no sangue 30 e 5 minutos depois da captura, respectivamente, independente do tempo de exposição aérea. Ambos os indicadores recuperaram os valores controle em 24 horas. Nos dois grupos de peixes, o cloreto plasmático diminuiu 60 minutos após captura e não recuperou os valores controle, enquanto o sódio sérico aumentou entre 15 e 30 minutos recuperando a condição controle em 24 horas. Não houve alteração significativa nos valores de potássio, cálcio, osmolaridade ou no hematócrito, hemoglobina, CV e VCM como consequência da captura. Os estressores sequenciais aplicados no pacu durante a captura ativaram o eixo cérebro-pituitária-interrenal (respostas do cortisol e glicose), mas a ativação do eixo cérebro-sistema simpático-células cromafins foi aparentemente moderada (respostas iônicas e hematológicas).

Análise da presença de agentes mutagênicos nas águas do Rio Pitimbú/RN

The Pitimbu River is located at the oriental portion of the State of Rio Grande do Norte, including three importants cities named Macaíba, Parnamirim e Natal. Although its high importance as a water source, which supplys great part of the South Zone of the Natal city, this river receives a large quantity of domestic and industrial waste water without treatment. The Pitimbu River headhas its river-head located in the city of Macaiba, goes through Parnamirim, then it flowing into at the Jiqui Lake in Natal. The aim of this study was to evaluate, qualitatively and quantitatively, the environmental quality of the Pitimbu River by genotoxicity bio-assays, which are important tools for genetics toxicological evaluation. In this work, five samples sites, distributed along the river, were used to collect water samples. Another point site, located near Jiqui lake, was used to collect drinkable water, which was treated by CAERN, the water treatment entreprise of Rio Grande do Norte. The following assays where used to evaluate the quality of these samples: Allium cepa assay; Comet assay; Micronuclei (MN) assay; and Ames test. For the Allium cepa assay, sixteen specimes where used for each water sample from the sample sites. In this assay both microscopic, like cytogenetic damage, and macroscopic aspects, as morphological variation were evaluated. Red blood cells from periferical blood of the Crenicichla menezesi native specie were used not only for the MN assay, but also for the Comet assay. These fishes were collected at different points on the Pitimbu River and the negative control was developed using fishes of the same species that were bring to the laboratory and maintained for 100 days in the optimal experimental conditions. For the Ames test, TA100, YG1042, TA98 and YG1041 strains were used in the directed method without metabolic activation. The results found by the Allium cepa assay showed that two water sample sites induced increase of mitotic index (IM). Additionally, compared to the control, all the water samples increased the chromossomal aberrations frequency and/or micronucleus. Among the sample sites, two also showed an abnormal growth rate in its root and two samples induced morphological alteration. With the MN test in red blood cells, a high frequence of MN was observed in tree sample. By comparing all the results obtained on the water sample points and with the negative control, a significant variation on the MN frequency was observed. Positive results were also observed for the same sample to water test by the Comet assay. These results allow concluding that the proposed specie Crenicichla menezesi has a good profile as a bio-indicator for the evaluation of environmental water quality and the MN and comet test can be usuful for in situ evaluation. By the Ames test, it was possible to detect the mutagenic activity on the waters from the Pitimbu River in different levels of mutagenicity. This result suggests that this river has several substances that induced changes directly to the DNA. The mechanisms involved to this phenomenon could be by both processes, by changing of the reading frame and by nucleotide substitution. These data set indicate the presence of mutagenic agents, which can represent in risk to biot and human beens

Consequências da colectomia associada à hepatectomia no metabolismo hepático e na forma e função de hemácias em ratos

This study investigated the influence of partial colectomy associated with hepatectomy on the biodistribution of the 99mTc-phytate, on metabolic parameters, as well as labeling and morphology of red blood cells. Wistar rats were distributed into three groups (each with 6), nominated as colectomy, colectomy+hepatectomy and sham. In the 30th postoperative day all rats were injected with 99mTc-phytate 0.1mL i.v. (radioactivity 0.66 MBq). After 15 minutes, liver sample was harvested and weighed. Percentage radioactivity per gram of tissue (%ATI/g) was determined using an automatic gamma-counter. Serum AST, ALT, alkaline phosphatase and red blood cells labeling were determined. The liver %ATI/g and red blood cells labeling were lower in colectomy and colectomy+hepatectomy rats than in sham rats (p <0.05), and no difference was detected comparing the colectomy and colectomy+hepatectomy groups. Red blood cells morphology did not differ among groups. Serum levels of AST, ALT and alkaline fosfatase were significantly higher in colectomy+hepatectomy than in colectomy rats (p<0.001). Hepatectomy associated with colectomy lowered the uptake of radiopharmaceutical in liver and in red blood cells in rats, coinciding with changes in liver enzymatic activity

Haematological characteristics of Brazilian Teleosts: III. Parameters of the hybrid tambacu (Piaractus mesopotamicus Holmberg x Colossoma macropomum Cuvier) (Osteichthyes, Characidae)

Fifty six specimens of the hybrid tambacu (Piaractus mesopotamicus Holmberg, 1887 male x Colossoma macropomum Cuvier, 1818 female) were collected from fishfarm of Guariba, São Paulo, to evaluate their haematology. Fishes presented 400.0 to 3,100.0 g total weight and 20.0 to 52.0 cm total length. Haemoglobin, haematocrit, mean corpuscular haemoglobin content (MCHC) and percentage of defense blood cells including leucocytes and thrombocytes, were studied. Statistical analysis showed positive correlation (P<0.01) between haematocrit, MCHC and haemoglobin rate. Nevertheless, thrombocytes and lymphocytes showed negative correlation (P<0.01).

Effect of Aloe vera extract on the improvement of the respiratory activity of leukocytes of matrinxã during the transport stress

This study evaluated the effect of extract of Aloe vera in the transport water of matrinxã (Brycon amazonicus) fish on stress response and leukocyte respiratory activity. Fish was transported for 4 h in water containing Aloe at levels 0; 0.02; 0.2 and 2 mg/L, and sampled before transport 2, 4, 24 and 96 h after for determination of plasma glucose and respiratory activity of leukocytes. An additional in vitro assay was conducted with another fish species, pacu (Piaractus mesopotamicus), to test the respiratory burst of leukocytes exposed to Aloe extract (0.0, phosphate-buffered saline (PBS) only) at 0.1, 0.2, 0.5 and 1 mg/L). Plasma glucose increased after 2 and 4 h of transport and returned to control levels within 24 h, but the addition of Aloe in the transport water did not affect the level of blood glucose. However, at 2 h of transport, Aloe enhanced the respiratory activity of leukocytes in a dose-dependent way. The highest value of respiratory burst activity of leukocytes was observed in the fish transported in water containing Aloe at 2 mg/L. The enhancing effect of the plant extract on the production of oxygen radicals was confirmed in vitro in leukocytes of pacu incubated in Aloe at concentrations 0.1 and 0.2 mg/L. The results suggest that Aloe vera is a modulator of the immune system in fish improving the innate immune response tested.

Influência da laparoscopia e da laparotomia na gasometria, leucometria diferencial e citocinas em modelo de sepse abdominal em ratos

Laparoscopic surgery is associated with reduced surgical trauma, and less acute phase response, as compared with open surgery. Cytokines are important regulators of the biological response to surgical and anesthetic stress. The aim of this study was to determine if CO2 pneumoperitoneum would change cytokine expression, gas parameters and leukocyte count in septic rats. Methods: Wistar rats were randomly assigned to five groups: control (anesthesia only), laparotomy, CO2 pneumoperitoneum, cecum ligation and puncture by laparotomy, and laparoscopic cecum ligation and puncture. After 30 min of the procedures, arterial blood samples were obtained to determine leukocytes subpopulations by hemocytometer. TNFα, IL-1β, IL-6 were determined in intraperitoneal fluid (by ELISA). Gas parameters were measured on arterial blood, intraperitoneal and subperitoneal exsudates. Results: Peritoneal TNFα, IL-1β and IL-6 concentrations were lower in pneumoperitoneum rats than in all other groups (p<0.05). TNFα, IL-1β and IL-6 expression was lower in the laparoscopic than in laparotomic sepsis (p<0.05). Rats from laparoscopic cecum ligation and puncture group developed significant hypercarbic acidosis in blood and subperitoneal fluid when compared to open procedure group. Total white blood cells and lymphocytes were significantly lower in laparoscopic cecum ligation and puncture rats than in the laparotomic (p<0.01). Nevertheless, the laparotomic cecum ligation rats had a significant increase in blood neutrophils and eosinophils when compared with controls (p<0.05). Conclusions: This study demonstrates that the CO2 pneumoperitoneum reduced the inflammatory and immune response in an animal model of peritonitis with respect to intraperitoneal cytokines, white blood cell count and clinical correlates of sepsis. The pneumoperitoneum produced hypercarbic acidosis in septic animals

Respostas hematológicas em tuvira após anestesia com diferentes concentrações de óleo de cravo

The hematological responses in tuvira (Gymnotus aff. inaequilabiatus) after clove oil anesthesia at different concentrations were evaluated in this study. Fish (n = 25) with average length of 26.8 +/- 2.4 cm and weighing 94.4 +/- 23.6 g were separated in five groups composed with five animals each and submitted individually to respective concentrations of clove oil: 40, 80, 120, 160 and 200 mg L-1. After anesthetic recuperation, the blood samples collection were performed, as well as complete hemogram. The results were submitted to analysis of variance, Tukey test (P<0.05) and Sperman's correlation analysis. There was an increase (P<0.05) on hematocrit, red blood cells count, thrombocytes count and in the values of neutrophils with increasing of clove oil concentrations. In addition, positive correlation (P<0.05) were observed on total leukocytes, as well as negative linear correlation (P<0.05) for mean corpuscular hemoglobin concentration (MCHC). The clove oil anesthesia on tuvira cause hematological changes with an effect dose-dependent. These changes should be considered mainly in future studies designed to establish the reference values of hematologic variables for this specie.

Parâmetros hematológicos de jumentas gestantes em diferentes períodos

The hematological parameters and total protein concentration in healthy donkeys during pregnancy, under grass handling conditions in tropical weather, were evaluated. Fifth-two Marchador Brasileiro breed healthy donkeys, ranging from 8 to 16 years old, were evaluated, 36 of which were pregnant. The animals were separated in four groups: non pregnant (control), pregnancy from 25 to 110 days (first phase), from 111 to 210 days (second phase), and from 211 to 340 days (third phase). Total protein, erythrocyte, and leukocytes counts; packed cell volume; hemoglobin concentration; hematimetric absolute rates of mean corpuscular volume (MCV); mean corpuscular hemoglobin (MCH); and mean corpuscular hemoglobin concentration (MCHC) were evaluated. Total protein and eosinophils count decreased in the first phase. Red blood cells, hemoglobin concentration, and MCHC increased in the third phase of pregnancy. However, MCV and MCH decreased in the same phase.

Leucogram and serum acute phase protein concentrations in queens submitted to conventional or videolaparoscopic ovariectomy

Trinta gatas, saudáveis, foram submetidas à ovariectomia pela técnica convencional e por videolaparoscopia. Amostras de sangue foram obtidas com o objetivo de verificar a intensidade da resposta inflamatória por meio da análise das concentrações de proteinas de fase aguda e contagem de leucócitos antes e até 144 horas após procedimento cirúrgico. As proteínas que apresentaram aumento significativo 24 horas após a cirurgia foram: ceruloplasmina, hemopexina, haptoglobina e α1-glicoproteína ácida, 69,8%, 103,5%, 117,3% e 199,0%, respectivamente, para ovariectomia convencional, e 22,3%, 46,1%, 79,8% e 74,6%, respectivamente, para ovariectomia por videolaparoscopia. A resposta inflamatória foi mais evidente nas gatas submetidas à ovariectomia convencional. Os resultados mostram aumento e diminuição na concentração de proteínas de fase aguda e na contagem de leucócitos, podendo ser utilizados na avaliação da resposta inflamatória induzida por procedimentos cirúrgicos.

Haematology of captive grey-brocket deer Mazama gouazoubira (Fischer, 1814) (Cervidae: Odocoileinae).

Reference intervals for haematologic and total plasma proteins profiles were determined for 11 physically restrained adult grey-brocket deer. Erythrocytic alterations, as discrete to moderate poikilocytosis, were observed in all animals. Red and white blood cells counts were determined by manual methodology. Obtained erythron and leukon values were slightly higher than previous reports. Statistical difference was not demonstrated between stags and hinds. No previous studies reported blood values for captive grey-brocket deer obtained by manual methods. Also, the authors suggest that future studies on deer haematology should be focused on technique evaluation and improvement, considering the unusual erythrocytic morphology.

Molecular diagnosis of Anaplasmataceae organisms in dogs with clinical and microscopical signs of ehrlichiosis

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