Mutagenic and carcinogenic potential of a textile azo dye processing plant effluent that impacts a drinking water source

Recently a textile azo dye processing plant effluent was identified as one of the sources of mutagenic activity detected in the Cristais River, a drinking water source in Brazil [G.A. Umbuzeiro, D.A. Roubicek, C.M. Rech, M.I.Z. Sato, L.D. Claxton, Investigating the sources of the mutagenic activity found in a river using the Salmonella assay and different water extraction procedures, Chemosphere 54 (2004) 1589-1597]. Besides presenting high mutagenic activity in the Salmonella/microsome assay, the mutagenic nitro-aminoazobenzenes dyes CI Disperse Blue 373, Cl Disperse Violet 93, and CI Disperse Orange 37 [G.A. Umbuzeiro, H.S. Freeman, S.H. Warren, D.P Oliveira, Y. Terao, T. Watanabe, L.D. Claxton, the contribution of azo dyes in the mutagenic activity of the Cristais river, Chemosphere 60 (2005) 55-64] as well as benzidine, a known carcinogenic compound [T.M. Mazzo, A.A. Saczk, G.A. Umbuzeiro, M.V.B. Zanoni, Analysis of aromatic amines in surface waters receiving wastewater from textile industry by liquid chromatographic with eletrochemical detection, Anal. Lett., in press] were found in this effluent. After similar to 6 km from the discharge of this effluent, a drinking water treatment plant treats and distributes the water to a population of approximate 60,000. As shown previously, the mutagens in the DWTP intake water are not completely removed by the treatment. The water used for human consumption presented mutagenic activity related to nitro-aromatics and aromatic amines compounds probably derived from the cited textile processing plant effluent discharge [G.A. Umbuzeiro, D.A. Roubicek, C.M. Rech, M.I.Z.. Sato, L.D. Claxton, Investigating the sources of the mutagenic activity found in a river using the Salmonella assay and different water extraction procedures, Chemosphere 54 (2004) 1589-1597; G.A. Umbuzeiro, H.S. Freeman, S.H. Warren, D.P. Oliveira, Y. Terao, T. Watanabe, L.D. Claxton, the contribution of azo dyes in the multagenic activity of the Cristais river, Chemosphere 60 (2005) 55-64]. Therefore, it is important to evaluate the possible risks involved in the human consumption of this contaminated water. With that objective, one sample of the cited industrial effluent was tested for carcinogenicity in the aberrant crypt foci medium-term assay in colon of Wistar rats. The rats received the effluent in natura through drinking water at concentrations of 0.1%, 1%, and 10%. The effluent mutagenicity was also confirmed in the Salmonella/microsome assay with the strains TA98 and YG1041. There was an increased number of preneoplastic lesions in the colon of rats exposed to concentrations of 1% and 10% of the effluent, and a positive response for both Salmonella strains tested. These results indicate that the discharge of the effluent should be avoided in waters used for human consumption and show the sensitivity of the ACF crypt foci assay as an important tool to evaluate the carcinogenic potential of environmental complex mixtures. (c) 2006 Elsevier B.V. All rights reserved.

Reversion to virulence evaluation of a 9R vaccine strain of Salmonella enterica serovar gallinarum in commercial brown layers

The live vaccine Cevac S. Gallinarum, made from a rough strain of Salmonella enterica subspecies enterica serotype Gallinarum is used for preventing fowl typhoid, a disease that still causes considerable economic losses in countries with a developing poultry industry. The objective of this paper was to evaluate a possible reversion to virulence of the strain used in a vaccine in commercial brown layers. Only Salmonella-free chicks were utilized. One hundred twenty (120) 12-day-old Dekalb brown layers divided in two trials were used. The first trial had six groups of 15 birds each. Birds of group 1 were vaccinated with 10 doses of Cevac S. Gallinarum subcutaneously and 10 doses orally, in a total of 20 doses of vaccine. Then the birds of groups 2, 3, 4, and 5 received inocula that contained feces and a pool of organs with fragments of liver, heart, spleen, and cecal tonsils obtained from the immediately previous group. The second trial had three groups with 10 birds each. Birds in group 7 received inocula containing a pool of organs from birds of group 5 from trial 1, whilst the birds in group 8 were vaccinated subcutaneously with one dose of vaccine. Both trials included negative control groups (6 and 9). Throughout the experimental period, birds were monitored for reactions to the vaccination on the site of administration, clinical signs, and post-mortem lesions. In each passage, in addition to the birds euthanized to provide the inocula material, two birds from each group were euthanized for assessment of possible lesions, and their organs (liver, heart, spleen and cecal tonsils) were cultured in an attempt to isolate the vaccine strain. Except for one bird from group 1, that had a local reaction on the site of vaccination - a small vesicle with less that 0.5 mm that persisted until the third day post vaccination -, no other bird had any local reaction to the vaccine or any visible clinical alteration. Birds in group 8 did not present any reaction or clinical alteration because of the vaccine. We only managed to re-isolate the vaccine strain in the inocula made from organs of birds in group 1. We confirmed the isolation by means of biochemical tests, serology, and acriflavine agglutination test. All other cultures made from organs or feces, from all the other experimental groups did not show any growth of the vaccine strain or any other Salmonella serovar, suggesting that the vaccinated birds did not shed the SG9R vaccine strain. No bird presented any clinical symptoms or died during the trials, and no gross lesions were observed in the post-mortem examinations. Under the controlled conditions and time-frame of the present experiment, it was possible to conclude that the rough 9R strain of Salmonella Gallinarum present in the vaccine Cevac S. Gallinarum (Ceva Campinas Ltda. - Campinas, SP - Brazil) did not revert to virulence.

Ultrasonographic monitoring of early pregnancy development in Murrah buffalo heifers (Bubalus bubalis)

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

Ovarian morphometric characterization and in vitro maturation of oocytes obtained from buffalo (Bubalus bubalis) ovaries - partial results

Buffalo ovaries were collected from a slaughterhouse (Frigol, Brazil) and transported to the laboratory in saline solution at 36 degrees C. The ovaries were dissected to realize the evaluations (weight, length, width and height of the ovary; corpus luteum and dominant follicle diameters). The Cumulus-oocyte complexes (COCs) were recovered by aspiration of 2-8 mm follicles. Selected COCs were matured in TCM 199 supplemented with 10% fetal bovine serum, sodium pyruvate, LH, FSH, estradiol and gentamicin. In vitro maturation was carried out at 38.5 degrees C for 22-24 h and 34-36 h. For the evaluation of the nuclear maturation the oocytes were placed in TCM 199 medium added with type v hialuronidase where the granulosa cells were extracted. The denuded oocytes were transferred to 10 mu l of Hoescht 33342 and the chromosomic configuration was evaluated. The oocytes were classified according to meiosis stage in: Germinal Vesicle, Germinal Vesicle Breakdown, Metaphase I, Metaphase II and Degenerated. The means of weight, length, width and height of the ovary were 3.83 g, 2.27 cm, 1.08 cm and 1.56 cm, respectively. The means of corpus luteum and dominant follicle diameters were 1.40 cm and 7.77 mm. The proportion of oocytes that reached metaphase II stage was: 36.68%.

Ultrasonographic evaluation of the conceptus from days 10 to 60 of pregnancy in jennies

Daily ultrasound examinations were conducted from Days 10 to 60 (ovulation = Day 0) of pregnancy to monitor the conceptus in jennies (n = 12). The embryonic vesicle was first detected on Day 11.5 +/- 0.9 (mean +/- SD; range 10 to 13d) and was mobile until movement ceased (fixation) on Day 15.5 +/- 1.4 (range, 13 to 18d). The vesicle was spherical from Days 10 to 18 (mean growth rate, 3.2 mm/d), non spherical (irregular) with a reduced growth rate (0.5 mm/d) from Days 19 to 29, and then grew at a moderate rate (1.6 mm/d) up to Day 46. on average, detection of the embryo proper (consistently located on the ventral aspect of the yolk sac) and embryonic heartbeat were Days 20.7 +/- 1.2 and 23.5 +/- 1.3, respectively. Formation of the allantoic sac was first detected on Day 24.4 +/- 1.7 and was complete on Day 36.8 +/- 1.6. Descent of the fetus (and formation of the umbilical cord) began on Day 37.9 +/- 1.7 and was complete on Day 44.1 +/- 2.1. Crown-ramp length averaged 3.7, 15.4, 22.7, 37.5 and 59.6 mm on Days 20, 30, 40, 50 and 60, respectively. In general, morphologic features and dates of occurrence were similar to those reported previously in the mare. (C) 1998 by Elsevier B.V.

Early embryonic death in mares: clinical and hormonal aspects

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

Cholinergic responses of seminal vesicles isolated from rats exposed perinatally to hydrocortisone

This study was performed in order to investigate the cholinomimetic response of seminal vesicles isolated from rats treated with hydrocortisone acetate during perinatal life. At the adult phase, the body weight and the wet weight of the seminal vesicle of these animals were unchanged. However, these male rats exhibited a significant reduction in plasma testosterone concentration. A significant increase in the sensitivity of the seminal vesicle to acetylcholine was also observed. Despite this, there was a significant reduction in the maximum contractile response of the organ to this transmitter. These results indicate that exposure to hydrocortisone during the critical period of brain sexual differentiation has a long-term effect on testosterone production of male rats. In addition, physiological levels of cortisone in perinatal life are also essential to support the contractile response pattern of the seminal vesicle to acetylcholine in adult life, probably crucial to the reproductive process. (C) 2003 Elsevier B.V. Ltd. All rights reserved.

Effects of prenatal hydrocortisone acetate exposure on fertility and sexual behavior in male rats

The aim of the present study was to investigate the effects of hydrocortisone during the prenatal period and its later repercussions on the fertility and sexual behavior of male rats. Pregnant rats were treated (s.c.) with hydrocortisone acetate, at 1.5 mg/day on the 17th, 18th, and 19th days of gestation. Decreased body weight and no alteration in anogenital distance were observed in male offspring. Adulthood, presented reductions of body weight, plasma testosterone levels, and seminal-vesicle wet weight without secretion as well as no alteration in the wet weights of the testes, epididymis, and seminal vesicle with secretion in the treated group. Males exposed to hydrocortisone during the prenatal period were able to mate with normal females, which became pregnant but exhibited an increased number of post-implantation losses. In spite of this, these treated males exhibited decreased male sexual behavior and the appearance of female sexual behavior after these male rats were castrated and pretreated with exogenous estrogen. These results indicate that exposure to hydrocortisone in the later stages of pregnancy may have a long-term effect on the fertility and sexual behavior of mate rats, suggesting an incomplete masculinization and defeminization of the central nervous system. (C) 2003 Elsevier B.V. (USA). All rights reserved.

In vivo uptake of a haem analogue Zn protoporphyrin IX by the human malaria parasite P. falciparum-infected red blood cells

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

Effect of follicle size on mRNA expression in cumulus cells and oocytes of Bos indicus: an approach to identify marker genes for developmental competence

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

A new genus and two new species of Stevardiinae (Characiformes: Characidae) with a hypothesis on their relationships based on morphological and histological data

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

A new vision of the origin and the oocyte development in the Ostariophysi applied to Gymnotus sylvius (Teleostei: Gymnotiformes)

Tendo por base os novos conhecimentos oriundos de recentes estudos com Perciformes marinho, a origem e o desenvolvimento dos oócitos no Ostariophysi Gymnotus sylvius são aqui descritos. da mesma maneira que ocorre nos Perciformes, em Gymnotus sylvius as oogônias são encontradas no epitélio germinativo que margeia as lamelas ovígeras. No início da foliculogênese, a proliferação das oogônias e sua entrada em meiose dão origem a ninhos de células germinativas que se projetam em direção ao estroma ovariano, a partir do epitélio germinativo. Os ninhos e o epitélio germinativo são suportados pela mesma membrana basal que os separa do estroma. Coincidindo com a paralisação da meiose os oócitos, presentes nos ninhos, são separados uns dos outros por processos citoplasmáticos das células pré-foliculares. As células pré-foliculares derivam do epitélio germinativo sendo, portanto, inicialmente células epiteliais. Durante a foliculogênese, ao mesmo tempo em que envolvem os oócitos individualizando-os, as células pré-foliculares sintetizam a membrana basal ao seu redor. Os oócitos entram em crescimento primário ainda dentro dos ninhos. Ao término da foliculogênese, o oócito e as células foliculares que compõem o folículo são circundados pela membrana basal. O folículo permanece conectado ao epitélio germinativo uma vez que ambos compartilham uma porção comum da membrana basal. Células oriundas do estroma circundam o folículo ovariano exceto na região de compartilhamento da membrana basal formando a teca. O folículo, a membrana basal e a teca formam o complexo folicular. O desenvolvimento do oócito ocorre dentro do complexo folicular e compreende os estágios de crescimento primário e secundário, maturação e ovulação. Os alvéolos corticais surgem no ooplasma momentos antes do início do crescimento secundário ou estágio vitelogênico que tem início com a deposição de vitelo, progride até o oócito esteja completamente desenvolvido e o ooplasma preenchido pelos glóbulos de vitelo. A maturação é caracterizada pela migração do núcleo ou vesícula germinativa, pela quebra da vesícula germinativa, ou seja, pela fragmentação do envoltório nuclear e, retomada da meiose. Na ovulação o ovo é liberado do complexo folicular para o lúmen ovariano. em comparação com os Perciformes marinhos com ovos pelágicos, o desenvolvimento oocitário em Gymnotus sylvius tem menos etapas dentro dos estágios de desenvolvimento, sendo as duas mais notáveis delas as ausências da formação das gotas de lipídio durante os crescimentos primário e secundário (e a consequente fusão das gotas para formar um único glóbulo de lipídio durante a maturação) e, a hidrólise do vitelo antecedendo a ovulação.

A new Moenkhausia Eigenmann, 1903 (Ostariophysi: Characiformes) from Chapada Diamantina, rio Paraguaçu Basin, Bahia, Northeastern Brazil

Moenkhausia diamantina, nova espécie, é descrita de tributário do rio Paraguaçu, BA, nordeste do Brasil. Esta espécie é distinguida das demais congêneres por caracteres relacionados ao padrão de coloração do corpo, pela presença de escamas na linha mediana pré-dorsal e número de raios da nadadeira anal.

Paternal treatment with cisplatin impairs reproduction of adult male offspring in rats

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Description of a new species of Pareiorhina (Siluriformes: Neoplecostominae) from Rio Sao Francisco basin

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