Mebendazole treatment against Anacanthorus penilabiatus (Monogenea, Dactylogyridae) gill parasite of cultivated Piaractus mesopotamicus (Osteichthyes, Characidae) in Brazil. Efficacy and hematology

The present work evaluated the efficacy of mebendazole (MBZ) treatment against infections with the monogenean helminths Anacanthorus penilabiatus, gill parasites of young cultivated pacu, Piaractus mesopotamicus. A short-term bath treatment using 100, 200 and 500 mg MBZ/1 of water for 10 and 30 min and a long-term bath using 1, 10 and 100 mg MBZ/1 of water for 24 h were utilized. Seven days after, fish were sacrificed and parasites counted. Concentrations of 500 and 200 mg MBZ/1 for 10 and 30 min showed reduced efficacy (0.0 and 0.7%) and (14.2 and 11.0%), respectively. Nevertheless, 100 mg MBZ/1 (10 min) and 10 mg MBZ/1 (24 h) showed better efficacy (79.6 and 81.4%, respectively). Treatments for 24 h provoked increases in hematocrit, hemoglobin concentration and leukocyte number when compared to those untreated fish. Mebendazole treatments also provoked alterations in the defense blood cells especially in lymphocyte and thrombocyte numbers, when the fishes were submitted to 10 and 30 min baths.

Características hematológicas, bioquímicas e biométricas de Piaractus mesopotamicus Holmberg, 1887 (Osteichthyes: Characidae) oriundos de cultivo intensivo

Hematological characteristics are important tools for diagnosis and prognostic features in fish population and they contribute to the comprehension of comparative physiology, phylogenetic relation, food habit and other ecological parameters. Red blood cell count (RBC), white blood cell count (WBC), thrombocytes counts, glucose, cortisol, hepatosomatic and splenosomatic relation were evaluated in Piaractus mesopotamicus. The values of hepatosomatic relation varied from 0.67% to 2.79% and splenosomatic relation from 0.011% to 0.108%. RBC 1.870 to 4.590 × 106/μL, hemoglobin concentration 5.7 to 12.1g/dL, haematocrit 24.0% to 40.0%, mean corpuscular volume 98.0 to 174.0 fL, mean corpuscular haemoglobin concentration 19.1 to 34.2g/dL, glucose 40.6 to 89.2mg/dL and cortisol 4.73 to 110.5ng/dL The number of total thrombocytes was 12,160.0 to 89,280.0/μL and WBC 2,019.0 to 47,473.0/μL. Differential count of leucocytes showed 54.1% limphocytes, 10.9% neutrophils, 10.3% monocytes, 9.9% PAS-positive granular leukocytes (PAS-GL), 6.7% eosinophils, 0.3% basophils and 7.8% immature leucocytes. Variation of these haematological characteristics in the same fish species was observed, being considered as the normal variation in the fish concerned. Hence it is incorrect to assume that biological characteristics in fishes are distributed normally.

Morphological, cytochemical, and ultrastructural study of thrombocytes and leukocytes in neotropical fish, Brycon orbignyanus Valenciennes, 1850 (Characidae, Bryconinae)

Morphological, cytochemical and ultrastructural studies are important to demonstrate the function of the blood cells, which is very little understood in teleosts. In peripheral blood of piracanjuba' Brycon orbignyanus, thrombocytes, lymphocytes, monocytes, neutrophils and heterophils were studied and characterized. Thrombocytes had a fusiform or oval shape with PAS-positive granules. Lymphocytes presented small size with sparse basophilic cytoplasm. Monocytes were large in size, presented basophilic cytoplasm that may be foamy or vacuolated, with non-specific esterase staining. The neutrophils presented lightly neutrophilic granule cytoplasm, with positivity for PAS and peroxidase. The heterophils were large in size, with eosinophilic and basophilic granules cytoplasm and PAS-positive. Transmission electron microscopy study demonstrated that the thrombocytes, lymphocytes and monocytes features were similar to other teleosts. In ultrastructural study only one type of neutrophils was observed. Cytochemical findings indicated that neutrophils and monocytes of B. orbignyanus may be involved in phagocytosis, and neutrophils play an important microbicidal role.

Assessment of DNA damage by extracts and fractions of Strychnos pseudoquina, a Brazilian medicinal plant with antiulcerogenic activity

Strychnos pseudoquina St. Hil. is a native plant of the Brazilian Savannah, used in popular medicine to treat a number of conditions. Since it contains large quantities of alkaloids with proven antiulcer activity, we tested the genotoxic potential of crude extracts and fractions containing alkaloids and flavonoids from the leaves of this plant, on Salmonella typhimurium and performed the micronucleus test on peripheral blood cells of mice treated in vivo. The results showed that the methanol extract of the leaves of S. pseudoquina is mutagenic to the TA98 (-S9) and TA100 (+S9, -S9) strains of Salmonella. The dichloromethane extract was not mutagenic to any of the tested strains. Fractions enriched with alkaloids or flavonoids were not mutagenic. In vivo tests were done on the crude methanol extract in albino Swiss mice, which were treated, by gavage, with three different doses of the extract. The highest dose tested (1800 mg/kg b.w.) induced micronuclei after acute treatment, confirming the mutagenic potential of the methanol extract of the leaves of S. pseudoquina. In high doses, constituents of S. pseudoquina compounds act on DNA, causing breaks and giving rise to micronuclei in the blood cells of treated animals. © 2006 Elsevier Ltd. All rights reserved.

The influence of dietary vitamin C and E supplementation on the physiological response of pirarucu, Arapaima gigas, in net culture

This study evaluated the efficacy of dietary vitamin C (ascorbic acid or AA), vitamin E (α-tocopherol or α-T), and C + E supplementation on the blood parameters of Arapaima gigas grown in net cages for 45 days. Four treatments were tested: control (commercial feed); C800; E500 and C + E (800 + 500) with supplementation of 800 mg AA kg- 1, 500 mg α-T kg- 1 and 800 + 500 mg AA + α-T kg- 1, respectively. Hematocrit (Ht), red blood cells (RBC), and hemoglobin concentration (Hb) (oxidative status indicators), thrombocytes and leukocytes (immunological indicators), plasma protein and glucose were evaluated. Fish fed vitamin C and C + E supplemented diets showed greater weight gain and survival. Dietary vitamin C and C + E diet supplementation resulted in increased Ht, Hb, RBC, MCHC, total leukocytes, total proteins, thrombocytes and eosinophils compared to the control and α-T. The α-tocopherol-supplemented diet reduced the number of total thrombocytes, lymphocytes and neutrophils and increased glucose and eosinophils relatively to the control. In general, leukocytes and thrombocytes were good indicators of the efficiency of vitamin on the defense mechanism of the A. gigas reared in cages. Results indicate that high α-T diet supplementation provides no benefit for the maintenance of the oxidative or the immunological status of A. gigas. However, it was demonstrated that high dietary AA improves A. gigas immunological status. Red blood cell indices and immune system indicators showed no synergistic effect between the vitamins after supplementing the A. gigas diet with α-T + AA. © 2006 Elsevier Inc. All rights reserved.

Valores da Amplitude de Distribuição do Tamanho dos Eritrócitos (RDW) em eqüinos Puro Sangue Inglês (PSI) submetidos a exercícios de diferentes intensidades

In order to evaluate the influence of exercise of different intensities on Red Blood Cell Distribution width (RDW) and Packed Cell Volume (VG) in Thoroughbred horses, blood was collected from 60 animals, 30 males and 30 females, subdivided in groups of horses with 24 to 36 months of age and not in training, and after 12 months of training, and horses with 36 to 48 months of age in training. Blood samples where collected before and after trot and gallop. Samples where analyzed with a automatic cell counter (Cell-Dyn 3500R, Abbott Diagnostic). Red Blood Cell Distribution width (RDW) values increased significantly after trot and gallop demonstrating a variation in the size of red blood cells, while Packed Medium Cell Volume (VGM) values did not show variations before or after exercise.

Antisperm antibodies in infertile men and their correlation with seminal parameters

Aim: Antisperm antibodies (ASA) in males cause the autoimmune disease 'immune infertility'. The present study intended to detect the presence of ASA and their incidence in men with unexplained infertility, as well as to evaluate the correlation between the presence of ASA and semen parameter alterations. Methods: Blood and sperm assessment were collected to carry out a direct and indirect mixed antiglobulin reaction (MAR) test and semen analysis in infertile and fertile men from the University Hospital of the Faculty of Medicine, Sao Paulo State University, Sao Paulo. Results: In the MARtest, 18.18% of infertile men were positive for ASA. In fertile men, no positivity was found. A significant correlation between the presence of ASA with an increased white blood cell count plus a decreased hypoosmotic swelling test result was observed. Conclusions: The results indicate that ASA are involved in reduced fertility. It is not ASA detection per.se that provides conclusive information about the occurrence of damage to fertility. The correlation between infertility and altered seminal parameters reinforce the ASA participation in this pathology. © 2007 The Authors Journal compilation. © 2007 Japan Society for Reproductive Medicine.

DNA damage and nitric oxide synthesis in experimentally infected Balb/c mice with Trypanosoma cruzi

This study aimed to evaluate whether experimental Chagas disease in acute phase under benznidazole therapy can cause DNA damage in peripheral blood, liver, heart, and spleen cells or induce nitric oxide synthesis in spleen cells. Twenty Balb/c mice were distributed into four groups: control (non-infected animals); Trypanosoma cruzi infected; T. cruzi infected and submitted to benznidazole therapy; and only treated with benznidazole. The results obtained with the single cell gel (comet) assay showed that T. cruzi was able induce DNA damage in heart cells of both benznidazole treated or untreated infected mice. Similarly, T. cruzi infected animals showed an increase of DNA lesions in spleen cells. Regarding nitric oxide synthesis, statistically significant differences (p < 0.05) were observed in all experimental groups compared to negative control, the strongest effect observed in the T. cruzi infected group. Taken together, these results indicate that T. cruzi may increase the level of DNA damage in mice heart and spleen cells. Probably, nitric oxide plays an important role in DNA damaging whereas benznidazole was able to minimize induced T. cruzi genotoxic effects in spleen cells. © 2006 Elsevier Inc. All rights reserved.

Assessment of the potential clastogenic/aneugenic risk of Casearia sylvestris extract using in vivo assays

Casearia sylvestris (Flacourtiaceae) is a plant which grows in wild and has been widely used in folk medicine. In this study, clastogenic/aneugenic properties of Casearia sylvestris crude ethanolic extract were evaluated using in vivo chromosomal aberrations (CAs) and micronucleus (MN) assays in rodents. The animals were treated by gavage with 3 concentrations of the extract: 150, 300 and 500 mg/kg body weight. Bone marrow cells from Wistar rats were collected 24 h after having been submitted to the MN and CAs test. Peripheral blood cells from Swiss mice were collected 48 and 72 h after having been submitted to the MN test. The results show that C. sylvestris extract does not induce a significant increase in mean values for micronucleated polychromatic erythrocytes (MNPCE) in Swiss mice and Wistar rats, or CAs in rat bone marrow cells, at the 3 tested doses, indicating that the extract showed no clastogenic/aneugenic effects on chromosomes of the rodent cells tested. © 2007 The Japan Mendel Society.

Metodologia para quantificação de leucóciotos totais em peixe Oreochromis Niloticus

Haematological investigation is an important part of disease diagnosis. The techniques used for mammals are generally applicable for fishes with slight modification. The presence of nucleated erythrocytes and thrombocytes in fish may cause some confusion in the identification of blood cells, mainly in total leukocytes count. This work evaluated two different methods (direct and indirect) of leukocytes counting. Specimens of tilapia were divided randomly into four groups of 48 fish each. Each group was further randomized into three replicate experiments of 16 fish per replicate in 40 L aquarium and maintained for 10 days. Counting of leukocytes in peripheral blood was determined in six fishes per group at the times: 0, 3, 7 and 10 days. The direct method in Neubauer chamber showed a large dispersion of data in regard to the average, the opposite was observed in indirect method determined in blood smears that showed more homogeneity among groups. Due to this factor the indirect method can be considered more accurate than the direct method for counting leukocytes in fishes.

Mitigating stress effects during transportation of matrinxã(Brycon amazonicus Günther, 1869; Characidae) through the application of calcium sulfate

This study verified the effects of CaSO4 on physiological responses of the tropical fish matrinxãBrycon amazonicus(200.2 ± 51.1 g) in water containing CaSO4 after a 4-h transportation at concentrations of: 0, 75, 150, and 300 mg L-1. Blood samples were collected prior to transportation (initial levels), immediately after packaging, at arrival, and 24 h and 96 h after transportation (recovery). Cortisol levels increased after ackaging (118.2 ± 14.2 ng ml-1), and decreased slightly after transportation in water containing CaSO4 (106.8 ± 14.1), but remained higher than initial levels (21.0 ± 2.6 ng ml)1). Fish kept at 150 mg L-1 CaSO4 reached the pre-transportation levels at 24 h of recovery. Blood glucose increased after transportation in all treatments (8.2 ± 0.2 mmol L-1) and declined after full recovery to values below initial levels (4.8 ± 0.1 mmol L-1). Chloride levels did not change in CaSO4 treatments; serum sodium concentrations decreased after packaging and after transportation. Serum calcium levels did not differ among treatments, but decreased after packaging and increased at 96 h of recovery. Hematocrit and the number of red blood cells were higher in all treatments after packaging and arrival, except in fish exposed to 300 mg L-1 CaSO4. Mean corpuscular volume increased in 75 mg L-1 CaSO4, which reached the higher VCM after transportation. Hemoglobin levels increased only after transportation, regardless of calcium sulfate levels. Handling before transportation and transportation itself were both stressful to fish; calcium sulfate at concentrations tested in the present work had a moderate influence in the reduction of stress responses. © 2009 Blackwell Verlag, Berlin.

Experimental infection of commercial layers using a Salmonella enterica serovar Gallinarum strain: Leukogram and serum acute-phase protein concentrations

The aim of the present study was to evaluate white blood cell counts and serum protein profiles of commercial layers experimentally infected with Salmonella Gallinarum (SG) in order to better understand the pathophysiology of the disease caused by this bacterium. 180 five-day-old commercial layers were divided into 3 groups (G); G1 and G2 received 0.2 mL of inoculate containing 3.3x10 8 CFU or 3.3×10 5 CFU SG resistant to nalidix acid (Nal r)/mL, respectively, directly into their crops. G3 group did not receive the inoculum. Birds were sacrificed 24 hours before (T1) and 24 hours after the infection (T2), and three (T3), five (T4), seven (T5), and ten (T6) days after the administration of the inoculum. White blood cell counts were carried out in a Neubauer hemocytometer and in blood smears. Serum protein concentrations, including acute-phase proteins, were determined using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Data were submitted to analysis of variance, and means were compared by Tukey's test (P <0.05). G1 and G2 groups presented higher leukocyte counts on T4 and T5, respectively, due to the increase of circulating lymphocytes and heterophils, with a significant difference relative to G3. In electrophoresis, an increase in the serum levels of ceruloplasmin, haptoglobin, and hemopexin and a decrease in transferrin, which are acute-phase proteins, was verified. IgA serum levels did not change; however, IgG concentration increased during the infection. In conclusion, the results provide information for the better understanding of the pathophysiology of fowl typhoid.

Female sex hormones mediate the allergic lung reaction by regulating the release of inflammatory mediators and the expression of lung E-selectin in rats

Background: Fluctuations of estradiol and progesterone levels caused by the menstrual cycle worsen asthma symptoms. Conflicting data are reported in literature regarding pro and anti-inflammatory properties of estradiol and progesterone.Methods: Female Wistar rats were ovalbumin (OVA) sensitized 1 day after resection of the ovaries (OVx). Control group consisted of sensitized-rats with intact ovaries (Sham-OVx). Allergic challenge was performed by aerosol (OVA 1%, 15 min) two weeks later. Twenty four hours after challenge, BAL, bone marrow and total blood cells were counted. Lung tissues were used as explants, for expontaneous cytokine secretion in vitro or for immunostaining of E-selectin.Results: We observed an exacerbated cell recruitment into the lungs of OVx rats, reduced blood leukocytes counting and increased the number of bone marrow cells. Estradiol-treated OVx allergic rats reduced, and those treated with progesterone increased, respectively, the number of cells in the BAL and bone marrow. Lungs of OVx allergic rats significantly increased the E-selectin expression, an effect prevented by estradiol but not by progesterone treatment. Systemically, estradiol treatment increased the number of peripheral blood leukocytes in OVx allergic rats when compared to non treated-OVx allergic rats. Cultured-BAL cells of OVx allergic rats released elevated amounts of LTB4 and nitrites while bone marrow cells increased the release of TNF-α and nitrites. Estradiol treatment of OVx allergic rats was associated with a decreased release of TNF-α, IL-10, LTB4 and nitrites by bone marrow cells incubates. In contrast, estradiol caused an increase in IL-10 and NO release by cultured-BAL cells. Progesterone significantly increased TNF- α by cultured BAL cells and bone marrow cells.Conclusions: Data presented here suggest that upon hormonal oscillations the immune sensitization might trigger an allergic lung inflammation whose phenotype is under control of estradiol. Our data could contribute to the understanding of the protective role of estradiol in some cases of asthma symptoms in fertile ans post-menopausal women clinically observed. © 2010 de Oliveira et al; licensee BioMed Central Ltd.

Evaluation of Agaricus blazei in vivo for antigenotoxic, anticarcinogenic, phagocytic and immunomodulatory activities

The development of various types of cancer results from the interaction among endogenous, environmental and hormonal factors, where the most notable of these factors is diet. The aim of the present study was to determine the antigenotoxic, anticarcinogenic, phagocytic and immunomodulatory activities of Agaricus blazei. The test antigenotoxicity (Comet Assay) and anticarcinogenic (Test of Aberrant Crypt Foci) assess changes in DNA and/or intestinal mucosa that correlate to cancer development. Tests of phagocytosis in the spleen and differential count in blood cells allow the inference of modulation of the immune system as well as to propose a way of eliminating cells with DNA damage. Supplementation with the mushroom was carried out under pre-treatment, simultaneous treatment, post-treatment and pre-treatment. +. continuous conditions. Statistical analysis demonstrated that the mushroom did not have genotoxic activity but showed antigenotoxic activity. Supplementation caused an increase in the number of monocytes and in phagocytic activity, suggesting that supplementation increases a proliferation of monocytes, consequently increasing phagocytic capacity especially in the groups pre-treatment, simultaneous and pre-treatment. +. continuous. The data suggest that A. blazei could act as a functional food capable of promoting immunomodulation which can account for the destruction of cells with DNA alterations that correlate with the development of cancer, since this mushroom was demonstrated to have a preventive effect against pre-neoplastic colorectal lesions evaluated by the aberrant crypt foci assay. According to these results and the literature, it is believed that supplementation with A. blazei can be an efficient method for the prevention of cancer as well as possibly being an important coadjuvant treatment in chemotherapy. © 2011 Elsevier Inc.

Doppler ultrasonography principles and methods of evaluation of the reproductive tract in mares

Background: Doppler ultrasonography is a non-invasive real time pulse-wave technique recently used for the transrectal study of the reproductive system hemodynamics in large animals. This technic is based in the Doppler Effect Principle that proposes the change in frequency of a wave for an observer (red blood cells) moving relative to the source of the respective wave (ultrasonic transducer). This method had showed to be effective and useful for the evaluation of the in vivo equine reproductive tract increasing the diagnostic, monitoring, and predictive capabilities of theriogenology in mares. However, an accurate and truthful ultrasonic exam requires the previous knowledge of the Doppler ultrasonography principles. Review: In recent years, the capabilities of ultrasound flow imaging have increased enormously. The current Doppler ultrasound machines offer three methods of evaluation that may be used simultaneously (triplex mode). In B-mode ultrasound, a linear array of transducers simultaneously scans a plane through the tissue that can be viewed as a two-dimensional gray-scale image on screen. This mode is primarily used to identify anatomically a structure for its posterior evaluation using colored ultrasound modes (Color or Spectral modes). Colored ultrasound images of flow, whether Color or Spectral modes, are essentially obtained from measurements of moving red cells. In Color mode, velocity information is presented as a color coded overlay on top of a B-mode image, while Pulsed Wave Doppler provides a measure of the changing velocity throughout the cardiac cycle and the distribution of velocities in the sample volume represented by a spectral graphic. Color images conception varies according to the Doppler Frequency that is the difference between the frequency of received echoes by moving blood red cells and wave frequency transmitted by the transducer. To produce an adequate spectral graphic it is important determine the position and size of the simple gate. Furthermore, blood flow velocity measurement is influence by the intersection angle between ultrasonic pulses and the direction of moving blood-red cells (Doppler angle). Objectively colored ultrasound exam may be done on large arteries of the reproductive tract, as uterine and ovary arteries, or directly on the target tissue (follicle, for example). Mesovarium and mesometrium attachment arteries also can be used for spectral evaluation of the equine reproductive system. Subjectively analysis of the ovarian and uterine vascular perfusion must be done directly on the corpus luteum, follicular wall and uterus (endometrium and myometrium associated), respectively. Power-flow imaging has greater sensitivity to weak blood flow and independent of the Doppler angle, improving the evaluation of vessels with small diameters and slow blood flow. Conclusion: Doppler ultrasonography principles, methods of evaluation and reproductive system anatomy have been described. This knowledge is essential for the competent equipment acquisition and precise collection and analysis of colored ultrasound images. Otherwise, the reporting of inconsistent and not reproducible findings may result in the discredit of Doppler technology ahead of the scientific veterinary community.