Cryopreservation of immature bovine oocytes with 1,2 propanediol

O objetivo deste trabalho foi avaliar o efeito de dois gradientes de concentrações de 1,2 propanediol (PROH) na criopreservação de ovócitos imaturos, envoltos em células do cumulus, de bovinos abatidos em matadouro. No tratamento 1, com uso de 1,6 M de PROH, os ovócitos foram desidratados em três etapas crescentes (0,53; 1,06 e 1,6 M), à temperatura ambiente e congelados em nitrogênio líquido. A descongelação foi realizada em água a 37ºC por 30 segundos e a reidratação em três etapas decrescentes (1,6; 1,06 e 0,53 M), acrescidos de 0,25 M de sacarose cada. No tratamento 2 foi utilizado 2,0 M de PROH, de maneira semelhante ao tratamento 1, com desidratação contendo 0,7; 1,4 e 2,0 M, e reidratação 2,0; 1,4 e 0,7 M. Após a reidratação, os ovócitos foram lavados em meio TCM 199 (Meio de Cultivo para Tecidos) e levados para maturação in vitro. O grupo controle foi constituído de ovócitos recém-colhidos. Os ovócitos foram colocados para maturar por 24 horas, em meio TCM 199 com 10% de SVC (Soro de Vaca em Cio) e FSH (Hormônio Estimulante de Folículo), em co-cultura com células da granulosa, com 5% CO2 no ar a 39ºC. Os resultados de maturação nuclear foram diferentes (P<0,05) entre os ovócitos criopreservados e controle, sendo 2,04%, 0% e 84% para o tratamento 1, 2 e controle, respectivamente.A study was carried out to investigate the ability of immature bovine cumulus-oocytes complexes to survive cryopreservation and undergo subsequent in vitro maturation in two different levels of propanediol (PROH). Treatment 1 consisted of subjecting oocytes to a gradual 3-step, concentration-driven dehydration with PROH (0.53; 1.06 and 1.6 M) at room temperature and subsequent freezing. Oocytes were thawed in water at 37ºC for 30 seconds and subjected to a gradual 3-step, concentration-driven rehydration with PROH (1.6; 1.06 and 0.53 M) in the presence of 0.25 M sucrose. Treatment 2 was conducted the same way, but PROH concentrations for dehydration were 0.7; 1.4 and 2.0 M and for rehydration were 2.0; 1.4 and 0.7 M. After rehydration, oocytes of both treatments were washed in TCM 199 medium followed by the in vitro maturation. The control consisted of nonfrozen oocytes. Oocytes of all groups were cultured in TCM 199 medium with 10% ECS and FSH, and 5% CO2 air in co-culture at 39ºC, for 24 hours. The in vitro maturation rate of cryopreserved oocytes were different for treatments and control. Metafase II was reached by 2.04%, 0% and 84% for treatment 1, 2 and control, respectively

Relative expression of mRNAs related to cavitation process in bovine embryos produced in vivo and in vitro

The objectives of this work were to identify and to evaluate possible differences on gene expression of aquaporins and Na/K-ATPases transcripts between embryos in vivo and in vitro produced. For each group, 15 blastocysts distributed in three pools were used for RNA extraction followed by amplification and reverse transcription. The resulting cDNAs were submitted to Real-Time PCR, using the GAPDH gene as endogenous control. It was not possible to identify AQP1 transcripts. Relative expression of AQP3 (1.33 ± 0.78) and AQP11 (2.00 ± 1.42) were not different in blastocysts in vitro and in vivo produced. Na/K-ATPase &#945;1 gene (2.25 ± 1.07) was overregulated whereas Na/K-ATPase &#946;2 transcripts 0.40 ± 0.30) did not differ among blastocysts produced in vitro from those produced in vivo. Transcripts for gene AQP1 are not present in bovine blastocysts. In vitro culture system does not alter expression of genes AQP3, AQP11 and Na/K-ATPase &#946;2 genes, however, it affects expression of Na/K-ATPase &#945;1.Os objetivos neste trabalho foram identificar e avaliar possíveis diferenças na expressão gênica de transcritos de Aquaporina e ATPases-Na/K presentes em embriões produzidos in vivo e in vitro. Para cada grupo, 15 blastocistos distribuídos em três conjuntos foram utilizados para a extração do RNA, seguida da amplificação e da transcrição reversa. Os DNAs complementares foram submetidos à reação em cadeia da enzima polimerase em tempo real, utilizando-se o gene GAPDH como controle endógeno. Não foi possível identificar transcritos de AQP1. A expressão relativa dos genes AQP3 (1,33 ± 0,78) e AQP11 (2,00 ± 1,42) não foi diferente em blastocistos produzidos in vitro e in vivo. O gene ATPase-Na/K &#945;1 (2,25 ± 1,07) encontrou-se sobrerregulado, enquanto o gene ATPase-Na/K &#946;2 (0,40 ± 0,30) não diferiu entre os blastocistos produzidos in vitro e aqueles produzidos in vivo. Transcritos para o gene AQP1 não estão presentes em blastocistos bovinos. O sistema de cultivo in vitro não influencia a expressão dos genes AQP3, AQP11 e ATPase-Na/K &#946;2, porém altera a expressão do gene ATPase-Na/K &#945;1

Luteal dynamics in goats: morphological and endocrine features

The aim of this study was to establish the morphologic and endocrine characteristics of luteal dynamics in goats. It was used Toggenburg female goats that showed natural estrus in a 48-hour interval. After estrus, ultrasonographic evaluations of the ovaries were daily performed during 21 days using a portable device (5MHz probe). Blood sample was collected for plasma progresterone (P4) determination. Corpora lutea were detected for the first time on day 5 and progressively increased in size until D9 (1.26 ± 0.08 cm²), with no variation on subsequent days. In females with one ovulation, the first visualization of the corpora lutea was earlier than in those with multiple ovulation (4.54 ± 0.18 vs 5.74 ± 0.25 days). At the moment of the first visualization, luteal area was smaller in animals with single ovulation. Plasma P4 concentration progressively increased until day 9 and it did not show significant increase until luteolysis, characterized by a sharp decrease in P4 concentration, reaching values below 1 ng/mL in 24 hours. The luteal area slowly and gradually decreased in size. It was observed a significant positive correlation between P4 concentration and area during luteogenesis and luteolysis (r = 0.63 and r = 0.50, respectively). When corpus luteum reached its maximum size (D9), female with more than one corpora lutea, with a greater luteal tissue area, did not show P4 concentration higher than those with one ovulation (5.92 ± 0.59 vs 7.04 ± 0.79 ng/mL). These results show that luteal dynamics in Toggenbur goats follow a similar pattern to those observed in other goat breeds and luteal tissue growth was positively correlated with corpora lutea functionality.Objetivou-se neste estudo estabelecer as características morfológicas e endócrinas da dinâmica luteal em cabras. Foram utilizadas fêmeas da raça Toggenburg que manifestaram estro natural em um intervalo de 48 horas. Após o estro, foram realizadas avaliações ultrassonográficas diárias dos ovários durante 21 dias, utilizando-se um aparelho portátil (5 MHz). Amostras de sangue foram coletadas para dosagem de progesterona (P4) no plasma. Os corpos lúteos foram detectados pela primeira vez no D5 e aumentaram progressivamente de tamanho até o D9 (1,26 ± 0,08 cm²), não havendo variação nos dias subsequentes. Nas fêmeas com uma ovulação, a primeira visualização do corpo lúteo foi mais precoce que naquelas com ovulação múltipla (4,54 ± 0,18 vs 5,74 ± 0,25 dias). No momento da primeira visualização, a área luteal foi menor nos animais com uma ovulação. A concentração plasmática de P4 aumentou progressivamente até o D9 e não apresentou aumento significativo até o momento da luteólise, caracterizada por uma acentuada queda da concentração de P4, atingindo valores inferiores a 1 ng/mL em um intervalo de 24 horas. A área luteal diminuiu de forma lenta e gradual. Foi observada uma correlação positiva significativa entre a área e a concentração de P4 durante a lutegêonese e a luteólise (r = 0,63 e r = 0,50; respectivamente). No dia em que o corpo lúteo atinge sua área máxima (D9), as fêmeas com mais de um corpo lúteo, com maior área de tecido luteal, não apresentaram concentração de P4 superior à daquelas com uma ovulação (5,92 ± 0,59 vs 7,04 ± 0,79 ng/mL). Esses resultados indicam que a dinâmica luteal em caprinos da raça Toggenburg segue padrões semelhantes aos observados em outras raças e em outras espécies e que o crescimento de tecido luteal refletiu positivamente na funcionalidade do corpo lúteo

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear un derstanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5–7 vast areas of the tropics remain understudied.8–11 In the American tropics, Amazonia stands out as the world’s most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepre sented in biodiversity databases.13–15 To worsen this situation, human-induced modifications16,17 may elim inate pieces of the Amazon’s biodiversity puzzle before we can use them to understand how ecological com munities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple or ganism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region’s vulnerability to environmental change. 15%–18% of the most ne glected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lostinfo:eu-repo/semantics/publishedVersio

The complete genome sequence of Chromobacterium violaceum reveals remarkable and exploitable bacterial adaptability

Chromobacterium violaceum is one of millions of species of free-living microorganisms that populate the soil and water in the extant areas of tropical biodiversity around the world. Its complete genome sequence reveals (i) extensive alternative pathways for energy generation, (ii) ≈500 ORFs for transport-related proteins, (iii) complex and extensive systems for stress adaptation and motility, and (iv) wide-spread utilization of quorum sensing for control of inducible systems, all of which underpin the versatility and adaptability of the organism. The genome also contains extensive but incomplete arrays of ORFs coding for proteins associated with mammalian pathogenicity, possibly involved in the occasional but often fatal cases of human C. violaceum infection. There is, in addition, a series of previously unknown but important enzymes and secondary metabolites including paraquat-inducible proteins, drug and heavy-metal-resistance proteins, multiple chitinases, and proteins for the detoxification of xenobiotics that may have biotechnological applications

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear understanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5,6,7 vast areas of the tropics remain understudied.8,9,10,11 In the American tropics, Amazonia stands out as the world's most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepresented in biodiversity databases.13,14,15 To worsen this situation, human-induced modifications16,17 may eliminate pieces of the Amazon's biodiversity puzzle before we can use them to understand how ecological communities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple organism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region's vulnerability to environmental change. 15%–18% of the most neglected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lost