Levedura seca integral na alimentação de vacas lactantes / Dry yeast for lactating dairy cows

Os efeitos da inclusão de levedura seca de cana-de-açúcar (Saccharomyces cerevisiae) na dieta de vacas lactantes foram avaliados com base no fornecimento de quatro dietas constituídas de diferentes níveis de substituição do farelo de soja pela levedura de cana-de-açúcar (0,00; 0,33; 0,67 e 1,00kg/kg de MS). O delineamento utilizado foi o quadrado latino 4 × 4. Não houve efeito dos tratamentos sobre o consumo de MS, MO, EE, FDNi e NDT, porém ocorreu redução linear no consumo de PB e FDNcp e aumento do consumo de CNF. Diferenças estatísticas não foram verificadas para os coeficientes de digestibilidade aparente da MS, MO, PB, CNF e NDT; no entanto os coeficientes do FDNcp e do EE decresceram linearmente. Não foi observado efeito sobre a produção e eficiência de síntese microbiana, utilização dos componentes nitrogenados e produção e composição do leite. Conclui-se que a levedura seca pode substituir 100% o farelo de soja na alimentação de vacas lactantes.

The effects of the inclusion of sugar cane dry yeast (Saccharomyces cerevisiae) in dairy cows feeding were evaluated based on the provision of four different diets with increasing proportions of sugar cane dry yeast (0.00; 0.33; 0.67 and 1.00kg/kg of DM). The design adopted for this research was the latin square, 4 × 4. It was observed that the levels of yeast inclusion did not affect the intake of DM, OM, EE, NDFi and TDN. However, the CP and NDFcp intake decreased linearly, whereas the NFC intake increased linearly with the increasing levels of yeast. No effect on the digestibility coefficient of DM, OM, CP, NDFcp and NCF was observed. Nevertheless, the EE digestibility coefficient linearly decreased with the yeast inclusion in the experimental diets. There was no treatment effect on the production and efficiency of microbial synthesis, usage of nitrogenous components, as well as milk production and composition. It was concluded that the dried yeast can fully replace the soybean meal in diets of dairy cows.

Protocol to cryopreserve and isolate nuclei from adipose tissue without dimethyl sulfoxide.

Cryopreservation injuries involve nuclear DNA damage. A protocol for cryopreserving and isolating adipocyte nuclei is proposed. Adipose tissue samples were directly analyzed (NoCRYO-0h), or stored at -196°C for 7 days without 10% dimethyl sulfoxide (DMSO) (CRYO-WO-DMSO) or with DMSO (CRYO-W-DMSO). To determine the effect of DMSO on cryopreservation treatment, adipose tissue samples were stored at 4°C for 24 h with 10% DMSO (NoCRYO-W-DMSO-24h) and without (NoCRYO-WO-DMSO-24h). Samples were processed in isolation buffer, and nuclear integrity was measured by flow cytometry. The coefficient of variation, forward scatter, side scatter, and number of nuclei analyzed were evaluated. Pea (Pisum sativum) was used to measure the amount of DNA. All groups contained similar amounts of DNA to previously reported values and a satisfactory number of nuclei were analyzed. CRYO-W-DMSO presented a higher coefficient of variation (3.19 ± 0.09) compared to NoCRYO-0h (1.85 ± 0.09) and CRYO-WO-DMSO (2.02 ± 0.02). The coefficient of variation was increased in NoCRYO-W-DMSO-24h (3.80 ± 0.01) compared to NoCRYO-WO-DMSO-24h (2.46 ± 0.03). These results relate DMSO presence to DNA damage independently of the cryopreservation process. CRYO-W-DMSO showed increased side scatter (93.46 ± 5.03) compared to NoCRYO-0h (41.13 ± 3.19) and CRYO-WO-DMSO (48.01 ± 2.28), indicating that cryopreservation with DMSO caused chromatin condensation and/or nuclear fragmentation. CRYO-W-DMSO and CRYO-WO-DMSO presented lower forward scatter (186.33 ± 9.33 and 196.89 ± 26.86, respectively) compared to NoCRYO-0h (322.80 ± 3.36), indicating that cryopreservation reduced nuclei size. Thus, a simple method for cryopreservation and isolation of adipocyte nuclei causing less damage to DNA integrity was proposed.

DNA elimination in embryogenic development of Pennisetum glaucum x Pennisetum purpureum (Poaceae) hybrids.

Interspecific hybridization between Napier grass (Pennisetum purpureum), which is widely grown in Brazil for cattle forage, and pearl millet (Pennisetum glaucum) has been used as a breeding strategy for the development of improved cultivars. However, the hybrid between these two species is sterile due to its triploid condition (2n = 3x = 21 chromosomes), which hinders its use in crop breeding programs. It is known that genomic alterations result from the hybridization process. In order to measure the loss of DNA during embryo development, we used flow cytometry to estimate the nuclear DNA content of triploid and tetraploid embryos produced by interspecific hybridization between Napier grass and pearl millet. The triploid and tetraploid hybrids had a mean DNA content of 4.99-4.87 and 5.25-4.84 pg, at 10 and 30 days after pollination, respectively. The mean reduction in DNA content was higher in the tetraploid hybrids. The flow cytometry results revealed progressive genomic instability in these triploid and tetraploid hybrids, with this instability causing significant alterations in the DNA content of the hybrids.

Genotoxicity of SPL (spent pot lining) as measured by Tradescantia bioassays.

Spent Pot Liner (SPL) is a solid waste product generated in the process of aluminum production. Tradescantia micronuclei (Trad-MN) and stamen hair mutation (Trad-SHM) bioassays are very useful tests to assess genotoxicity of environmental pollutants. In the present study, we intended to investigate the genotoxicity of this waste with Tradescantia bioassays using leachates of SPL simulating the natural leachability of SPL in soil. The formation of micronuclei (MN) was found to be concentration dependent. MN frequency enhanced significantly with SPL treatment. In addition, SPL also appeared to increase the percentage of dyads and triads. Trad-SHM assay showed that SPL increases pink mutation events as SPL concentration increases. These results demonstrated that SPL is a cytogenotoxic agent that affects different genetic end-points (induction of micronuclei and point mutations) even at low concentration (2% and 3%).

Spent Pot Liner (SPL) induced DNA damage and nuclear alterations in root tip cells of Allium cepa as a consequence of programmed cell death.

There are various toxic effects of environmental pollutants, including apoptosis and carcinogenesis. Spent Pot Liner (SPL) is solid waste from the aluminum industry. It has a highly variable composition, including cyanide, fluoride, organics and metals. Preliminary characterizations of the effect of SPL on Allium cepa show the presence of condensed nuclei. Thus, the aim of this study was to analyze the toxic effect of SPL in A. cepa root meristem in the context of programmed cell death (PCD). A lot of specific features of this process such as DNA fragmentation, condensed chromatin, spherical nuclei and the formation of apoptotic-like bodies were observed in root meristem after SPL treatment. Root meristem treated with SPL 25% solution exhibited an alteration in antioxidant enzyme activities; a reduction in NCR as a consequence of high percentage of condensed nuclei; DNA fragmentation, detected by electrophoresis and TUNEL assay; cytoplasm vacuolization and also a disturbance in root morphology. These features are associated with programmed cell death (PCD) under abiotic stress. Therefore, these data show that SPL induces apoptosis-like PCD in root meristem cells of A. cepa.

Cytogenetic alterations induced by SPL (spent potliners) in meristematic cells of plant bioassays.

Spent potliners (SPL) is solid waste generated by the aluminum industry during the manufacture of aluminum metal. Information on the cytotoxicity effect of SPL is necessary to facilitate understanding of their action on organisms and to subsidize environmentally correct solutions. Thus, the aim of the present investigation is to compare the effect of SPL on meristematic cells of Allium cepa and Zea mays and also to discuss the mechanisms of SPL cytotoxicity involved. A strong inhibition on root growth in higher SPL concentrations has been observed in both A. cepa and Z. mays. For cytogenetic analysis, the results showed a reduction of mitotic index and increase of different abnormalities as the SPL concentration increased. We observed bridges, chromosome fragments, stickiness, multipolar anaphase, later segregation and cell death. In general, it was possible to observe an increase of different abnormalities as the SPL concentration increased. It is obvious from the results of the present investigation that SPL is cytotoxic on meristematic cells of plant tests (A. cepa and Z. mays).