Diversidade e aspectos bioecológicos de simulídeos (Diptera: Simuliidae) que ocorrem nos municípios adjacentes a construção da usina Hidrelétrica de Peixe-Angical, TO, Brasil

O objetivo desta Tese é realizar o levantamento taxonômico e avaliar a dinâmica populacional de Simuliidae em localidades sob influência do Aproveitamento Hidrelétrico de Peixe Angical, TO. Os simulídeos possuem abrangente distribuição geográfica, e os estágios imaturos utilizam ambientes lóticos como sítios de criação. Algumas espécies podem atuar como vetores de vírus, protozoários e helmintos, o que confere ao grupo importância médica e veterinária. O hábito hematofágico das fêmeas de simulídeos pode acarretar sérios prejuízos ao turismo; ocasiona baixa no rendimento escolar; e na agropecuária dificulta a execução do trabalho o que reduz a produtividade. Durante a construção de grandes empreendimentos ocorre em pouco tempo à introdução de contingente populacional com drástica transformação do meio. A intervenção do homem sobre os ecossistemas e o crescimento desordenado pode provocar desequilíbrio ecológico que propicia a proliferação de espécimes vetores com consequentes problemas médico sanitários. A maior parte dos trabalhos realizados com insetos vetores em áreas sob influência da construção de hidrelétricas se refere aos culicídeos. O estudo dos aspectos taxonômicos permitirá o levantamento da biodiversidade e o diferencial deste projeto está no estabelecimento da sazonalidade e dinâmica das populações de imaturos e adultos de simulídeos. As amostras foram obtidas em áreas de influência direta e indireta da UHE Peixe no rio Tocantins, em 12 pontos diferentes de coleta, nos municípios deJaú do Tocantins, Peixe, Palmeirópolis, Paranã e São Salvador do Tocantins. Foram realizadas bimestralmente de 2004 a 2007, um total de 24 campanhas para coleta em criadouros pré-selecionados, que acompanharam todas as fases de construção início das obras, formação do lago, funcionamento da Usina. OS dados abióticos foram aferidos, e os imaturos removidos do substrato manualmente por 10 minutos e posteriormente preparados para eclosão dos adultos. Parte do material foi identificado no Laboratório de Simulídeos e Oncocercose do Instituto Oswaldo Cruz, onde foram verificados novos registros específicos para a ocorrência de Simuliidae em Tocantins, além do assinalamento de espécies antropofílicas e/ou vetores de Onchocerca volvulus. Nas áreas usadas para a formação do lago houve desaparecimento de criadouros. O desmatamento ocorrido aliado ao vigor dos simulídeos que conseguem realizar voos de longas distâncias na procura de alimento ou locais adequados a oviposição devem ter contribuído para a dispersão de espécimes. Há relatos sobre a da ocorrência de oncocercose na área estudada, um foco foi demarcado na divisa de Goiás com Tocantins, municípios Paranã e Minaçu investigado a partir de um caso autóctone de oncocercose. Este estudo é relevante uma vez que o Brasil possui potencial hidroenergético e prevê a construção de inúmeras hidrelétricas nos próximos anos. É importante estudas as áreas impactadas, conhecer a sua biodiversidade e os aspectos bioecológicos de Simuliidae no país.

Compliance Program report of findings: FY 79 pesticides and metals in Fish Program (7305.007)

Because fish bioaccumulate* certain chemicals, levels of chemical contaminants in their edible portion must be closely monitored. In recent years, FDA has conducted several surveys designed to determine the occurrence and levels of selected chemicals or groups of chemicals in fish. Previous fish surveillance programs included the Mercury in Wholesale Fish Survey (FY 71), the FY 73 and 74 Comprehensive Fish Surveys, the Canned Tuna Program (FY 75), the Kepone and Mirex Contamination Program (FY 77), and the FY 77 Mercury in Swordfish Program. In addition, recent Compliance Programs for Pesticides and Metals in Foods and Pesticides, Metals, and Industrial Chemicals in Animal Feed have specified coverage of fish and fish products. Because of previous findings and the sustained high level of fish imported into the United States, a separate compliance program dealing solely with chemical contaminants in fish was initiated by the FDA Bureau of Foods in FY 78. The program includes all domestic and imported fish coverage except that directed by the Bureau of Veterinary Medicine for animal feed components derived from fishery products. The earlier surveys indicated that "bottom feeder" species such as catfish generally had the highest levels of pesticides and polychlorinated biphenyls (PCBs). For this reason, coverage at these species has been emphasized. Similarly, tuna has received special attention because it is the most prevalent fish in the U.S. diet and because of potential problems with mercury. Halibut, swordfish, and snapper also were emphasized in the sampling because of potential problems with mercury levels determined in previous years. The findings in this program were used in detecting emerging problems in fish and directing FDA efforts to deal with them. Care must be exercised in drawing conclusions about trends from the data because this Compliance Program was not statistically designed. Sampling objectives and sources may vary from year to year; thus the results are not directly comparable.

Development, validation, and utilization of a competitive enzyme-linked immunosorbent assay for the detection of antibodies against Brucella species in marine mammals

A competitive enzyme-linked immunosorbent assay (cELISA) was developed by using a whole-cell antigen from a marine Brucella sp. isolated from a harbor seal (Phoca vitulina). The assay was designed to screen sera from multiple marine mammal species for the presence of antibodies against marine-origin Brucella. Based on comparisons with culture-confirmed cases, specificity and sensitivity for cetacean samples tested were 73% and 100%, respectively. For pinniped samples, specificity and sensitivity values were 77% and 67%, respectively. Hawaiian monk seal (Monachus schauinslandi; n = 28) and bottlenose dolphin (Tursiops truncatus; n = 48) serum samples were tested, and the results were compared with several other assays designed to detect Brucella abortus antibodies. The comparison testing revealed the marine-origin cELISA to be more sensitive than the B. abortus tests by the detection of additional positive serum samples. The newly developed cELISA is an effective serologic method for detection of the presence of antibodies against marine-origin Brucella sp. in marine mammals.

Bartonella species detection in captive, stranded and free-ranging cetaceans

We present prevalence of Bartonella spp. for multiple cohorts of wild and captive cetaceans. One hundred and six cetaceans including 86 bottlenose dolphins (71 free-ranging, 14 captive in a facility with a dolphin experiencing debility of unknown origin, 1 stranded), 11 striped dolphins, 4 harbor porpoises, 3 Risso's dolphins, 1 dwarf sperm whale and 1 pygmy sperm whale (all stranded) were sampled. Whole blood (n = 95 live animals) and tissues (n = 15 freshly dead animals) were screened by PCR (n = 106 animals), PCR of enrichment cultures (n = 50 animals), and subcultures (n = 50 animals). Bartonella spp. were detected from 17 cetaceans, including 12 by direct extraction PCR of blood or tissues, 6 by PCR of enrichment cultures, and 4 by subculture isolation. Bartonella spp. were more commonly detected from the captive (6/14, 43%) than from free-ranging (2/71, 2.8%) bottlenose dolphins, and were commonly detected from the stranded animals (9/21, 43%; 3/11 striped dolphins, 3/4 harbor porpoises, 2/3 Risso's dolphins, 1/1 pygmy sperm whale, 0/1 dwarf sperm whale, 0/1 bottlenose dolphin). Sequencing identified a Bartonella spp. most similar to B. henselae San Antonio 2 in eight cases (4 bottlenose dolphins, 2 striped dolphins, 2 harbor porpoises), B. henselae Houston 1 in three cases (2 Risso's dolphins, 1 harbor porpoise), and untyped in six cases (4 bottlenose dolphins, 1 striped dolphin, 1 pygmy sperm whale). Although disease causation has not been established, Bartonella species were detected more commonly from cetaceans that were overtly debilitated or were cohabiting in captivity with a debilitated animal than from free-ranging animals. The detection of Bartonella spp. from cetaceans may be of pathophysiological concern.

Chromosomal rearrangements underlying karyotype differences between Chinese pangolin (Manis pentadactyla) and Malayan pangolin (Manis javanica) revealed by chromosome painting

The Chinese pangolin (Manis pentadactyla), a representative species of the order Pholidota, has been enlisted in the mammalian whole-genome sequencing project mainly because of its phylogenetic importance. Previous studies showed that the diploid number o

NONINVASIVE GIANT PANDA PATERNITY EXCLUSION

Giant panda hair samples obtained by noninvasive methods served as a source of DNA for amplification of seven giant panda microsatellite loci utilizing the polymerase chain reaction. Thirteen giant pandas held in Chinese zoos were tested for identification of paternity. Some males listed as sires have been excluded as the biological father of captive-born giant pandas. Because of the death of some potential sires, paternity is still not assigned for some giant pandas, although there is a high likelihood that paternity assignment could be made if postmortem samples are available for genetic analysis. The DNA microsatellite variation assayed by the test we have developed provides a rapid, highly informative, and noninvasive method for paternity identification in giant pandas. (C) 1994 Wiley-Liss, Inc.

Extraction, PCR amplification, and sequencing of mitochondrial DNA from scent mark and feces in the giant panda

To expand the feasibility of applying simple, efficient, non-invasive DNA preparation methods using samples that can be obtained from giant pandas living in the wild, we investigated the use of scent markings and fecal samples. Giant panda-specific oligonucleotide primers were used to amplify a portion of the mitochondrial DNA control region as well as a portion of the mitochondrial DNA cytochrome b gene and tRNA(Thr) gene region. A 196 base pair (bp) fragment in the control region and a 449 bp fragment in the cytochrome b gene and tRNA(Thr) gene were successfully amplified. Sequencing of polymerase chain reaction (PCR) products demonstrated that the two fragments are giant panda sequences. Furthermore, under simulated field conditions we found that DNA can be extracted from fecal samples aged as long as 3 months. Our results suggest that the scent mark and fecal samples are simple, efficient, and easily prepared DNA sources. (C) 1998 Wiley-Liss, Inc.

SEROLOGICAL SURVEY OF A CAPTIVE MACAQUE COLONY IN CHINA FOR ANTIBODIES TO SIMIAN TYPE-D RETROVIRUSES

Sera from 510 macaques consisting of Macaca mulatta, Macaca assamensis, Macaca fascicularis, Macaca nemestrina, and Macaca arctoides were investigated for antibodies to simian AIDS type D retrovirus (SRV) by ELISA and Western blot with viral antigens purified from supernatants of SRV-1 infected cell cultures. Of these monkeys, 104 were seropositive by ELISA; only 23 were confirmed by Western blot. The true positive reaction to SRV was found in 15 of 463 (3.2%) M. mulatta and eight of eleven (72.7%) M. assamensis.

Effects of rhFSH regimen and time interval on ovarian responses to repeated stimulation cycles in rhesus monkeys during a physiologic breeding season

We studied the effects of repeated stimulation by recombinant human FSH (rhFSH) at various time intervals during a physiologic breeding season in rhesus monkeys. Ovarian recovery and responses were assessed by ultrasonography, serum steroid concentrations

Dynamic changes in ovarian follicles measured by ultrasonography during gonadotropin stimulation in rhesus monkeys

The objective was to study dynamic changes of ovaries in rhesus macaques stimulated by gonadotropins to identify an indicator for predicting ovarian response to stimulation. Twenty-one cycling monkeys were given 36 IU/d recombinant human follicle-stimulat

Ovarian response to gonadotropin stimulation in juvenile rhesus monkeys

The objective of this study was to investigate juvenile rhesus monkeys responding to various gonadotropin regimen Stimulations. Thirty-two prepubertal rhesus monkeys were randomly allocated into five groups for ovarian stimulation as follows: Groups I, II

Cryopreservation of Kunming mouse oocytes using slow cooling, ultrarapid cooling and vitrification protocols

The cryopreservation of oocytes has been only marginally successful with any of the current protocols, including slow cooling, rapid cooling and vitrification. We wished to test the hypothesis that oocytes from a single mouse strain would freeze successfully by 1 of the 3 mentioned protocols. Unfertilized Kunming mouse oocytes obtained 14 h after PMSG/hCG administration were randomly assigned to be cryopreserved after slow cooling, ultra rapid cooling and vitrification. Oocytes were thawed by straws being placed into 37 degrees C water, and their morphological appearance and in vitro fertilization capability were compared with that of oocytes that had not undergone cryopreservation. Survival of oocytes was indicated by the absence of darkened ooplasm or by broken membranes or zona pellucida. Functional integrity was evaluated by the formation of a 2-cell embryo after IVF. Survival rate of slow cooled oocytes did not differ from that seen in vitrified oocytes (55.1 vs 65.9%) but was significantly lower in the rapidly cooled oocytes (24.2%; P<0.01). The results of NF of slow cooled and vitrified oocytes were similar to those of the control group (72 and 73 vs 77%; P>0.05). It appears that Kunming mouse oocytes can be successfully cryopreserved using the slow cooling method with 1,2-propanediol and vitrification, which contains both permeating and nonpermeating cryoprotectants. (C) 1997 by Elsevier Science Inc.

Development of zona-free hamster ova to blastocysts in vitro

The zona pellucida (ZP) enclosing the mammalian ovum is important for its protection and for initial stages of fertilization, but the role of the ZP during embryo development is less clear. This study was designed to investigate if the hamster ZP is needed for embryo development from 1-cell to blastocyst in vitro, and to compare methods for removing the ZP. A total of 395 hamster pronucleate ova were collected 10 h post activation from superovulated, mated female hamsters. The ZP was removed from some ova using either 0.05% pronase, 0.05% trypsin or acid Tyrode's solution. To prevent ZP-free ova from sticking together, they were cultured singly in 30-50 muL drops of HECM-6 culture medium together with ZP-intact ova as controls. There was no significant difference among treatment groups in embryo development to blastocyst: 36/87 (42%) in the ZP intact group; 35/75 (47%) in the pronase-treated ZP-free group; 37/74 (50%) in the trypsin-treated ZP-free group; and 37/71 (52%) in the acid-treated ZP-free group. These results indicate that 1) the ZP is unnecessary for hamster embryo development in vitro from the pronucleate ovum stage to blastocyst; 2) none of the three ZP-removal methods was detrimental to embryo development; 3) embryos do not need to be cultured in groups during in vitro development from 1-cell to blastocyst. (C) 2000 by Elsevier Science Inc.

Effect of antibiotics on development in vitro of hamster pronucleate ova

Antibiotics are commonly added to embryo culture media, but effects on embryo development have not been examined thoroughly. Hamster ova were used to investigate whether penicillin, streptomycin or gentamicin affect embryo development in vitro. Ova were collected 10 h post activation by spermatozoa in vivo and cultured in five treatments: 1) Control: chemically-defined medium HECM-9 with no antibiotics; 2) HECM-9 with 100 IU/mL, penicillin; 3) HECM-9 with 50 mug/mL streptomycin; 4) HECM-9 with 10 mug/mL,gentamicin and 5) HECM-9 with both 100 IU/mL penicillin and 50 mug/mL streptomycin. Individually, penicillin, streptomycin and gentamicin did not affect embryo development to the 8-cell stage at 58 h post oocyte activation, or morula/blastocyst stages, or blastocysts alone at 82 h post activation. However, when penicillin and streptomycin were both present in the culture medium the percentages of 8-cell embryos at 58 h and blastocysts at 82 h were significantly lower than the control. No antibiotic treatment improved hamster embryo development in vitro. We caution against the use of penicillin and streptomycin together for hamster embryo culture, and show that it is not necessary to include any antibiotics in embryo culture media for up to 72 h if proper sterile technique is used with an oil overlay. (C) 2000 by Elsevier Science Inc.