Ecosystem responses to fire : identifying cross-taxa contrasts and complementarities to inform management strategies

Changes in fire frequency, extent, and intensity mean that understanding the effects of fire on plants and animals is a primary concern for ecologists and land managers. Given the potentially conflicting fire responses of species both within and across taxonomic groups, prescribing fire regimes based on the response of one or only a few species may have negative consequences for other species. Here, we integrate data collected from a series of independent but complementary studies spanning a 75 + year chronosequence in a semi-arid shrubland ecosystem in south-western Australia to consider how fire management can best promote biodiversity both within and across taxonomic groups (plants, birds, small mammals, and reptiles). Younger fire ages (6–14 years) contained sparse shrubs, large areas of bare ground, and lacked a distinct litter layer and canopy. The oldest vegetation (60–85 years) had a distinct canopy, a well-developed litter layer and cryptogamic crust, higher variability in patch width, and more woody debris. Plant species richness and diversity decreased with time since fire, whereas bird species richness and diversity increased with time since fire, and mammal and reptile species richness and diversity showed no trend. The composition of all four taxonomic groups varied according to time since fire and the presence of 11 species was confined above or below certain fire-age thresholds. Our results support the need to maintain a mix of both younger and older fire ages across the landscape to maximise species diversity, and highlight the particular importance of older fire ages for many species. Future fire management for biodiversity conservation will benefit from identifying and reconciling cross-taxa contrasts and complementarities.

The role of inflammation in vascular function and pregnancy outcome in the pregnant stroke prone spontaneously hypertensive rat

During pregnancy, the maternal cardiovascular system undergoes major adaptation. One of these changes is a 40-50 % increase in circulating blood volume which requires a systemic remodelling of the vasculature in order to regulate maternal blood pressure and maximise blood supply to the developing placenta and fetus. These changes are broadly conserved between humans and rats making them an appropriate pre-clinical model in which to study the underlying mechanisms of pregnancy-dependent cardiovascular remodelling. Whilst women are normally protected against cardiovascular disease; pregnancy marks a period of time where women are susceptible to cardiovascular complications. Cardiovascular disease is the leading cause of maternal mortality in the United Kingdom; in particular hypertensive conditions are among the most common complications of pregnancy. One of the main underlying pathologies of these pregnancy complications is thought to be a failure of the maternal cardiovascular system to adapt. The remodelling of the uterine arteries, which directly supply the maternal-fetal interface, is paramount to a healthy pregnancy. Failure of the uterine arteries to remodel sufficiently can result in a number of obstetric complications such as preeclampsia, fetal growth restriction and spontaneous pregnancy loss. At present, it is poorly understood whether this deficient vascular response is due to a predisposition from existing maternal cardiovascular risk factors, the physiological changes that occur during pregnancy or a combination of both. Previous work in our group employed the stroke prone spontaneously hypertensive rat (SHRSP) as a model to investigate pregnancy-dependent remodelling of the uterine arteries. The SHRSP develops hypertension from 6 weeks of age and can be contrasted with the control strain, the Wistar Kyoto (WKY) rat. The phenotype of the SHRSP is therefore reflective of the clinical situation of maternal chronic hypertension during pregnancy. We showed that the SHRSP exhibited a deficient uterine artery remodelling response with respect to both structure and function accompanied by a reduction in litter size relative to the WKY at gestational day (GD) 18. A previous intervention study using nifedipine in the SHRSP achieved successful blood pressure reduction from 6 weeks of age and throughout pregnancy; however uterine artery remodelling and litter size at GD18 was not improved. We concluded that the abnormal uterine artery remodelling present in the SHRSP was independent of chronic hypertension. From these findings, we hypothesised that the SHRSP could be a novel model of spontaneously deficient uterine artery remodelling in response to pregnancy which was underpinned by other as yet unidentified cardiovascular risk factors. In Chapter 1 of this thesis, I have characterised the maternal, placental and fetal phenotype in pregnant (GD18) SHRSP and WKY. The pregnant SHRSP exhibit features of left ventricular hypertrophy in response to pregnancy and altered expression of maternal plasma biomarkers which have been previously associated with hypertension in human pregnancy. I developed a protocol for accurate dissection of the rat uteroplacental unit using qPCR probes specific for each layer. This allowed me to make an accurate and specific statement about gene expression in the SHRSP GD18 placenta; where oxidative stress related gene markers were increased in the vascular compartments. The majority of SHRSP placenta presented at GD18 with a blackened ring which encircled the tissue. Further investigation of the placenta using western blot for caspase 3 cleavage determined that this was likely due to increased cell death in the SHRSP placenta. The SHRSP also presented with a loss of one particular placental cell type at GD18: the glycogen cells. These cells could have been the target of cell death in the SHRSP placenta or were utilised early in pregnancy as a source of energy due to the deficient uterine artery blood supply. Blastocyst implantation was not altered but resorption rate was increased between SHRSP and WKY; indicating that the reduction in litter size in the SHRSP was primarily due to late (>GD14) pregnancy loss. Fetal growth was not restricted in SHRSP which led to the conclusion that SHRSP sacrifice part of their litter to deliver a smaller number of healthier pups. Activation of the immune system is a common pathway that has been implicated in the development of both hypertension and adverse pregnancy outcome. In Chapter 2, I proposed that this may be a mechanism of interest in SHRSP pregnancy and measured the pro-inflammatory cytokine, TNFα, as a marker of inflammation in pregnant SHRSP and WKY and in the placentas from these animals. TNFα was up-regulated in maternal plasma and urine from the GD18 SHRSP. In addition, TNFα release was increased from the GD18 SHRSP placenta as was the expression of the pro-inflammatory TNFα receptor 1 (Tnfr1). In order to investigate whether this excess TNFα was detrimental to SHRSP pregnancy, a vehicle-controlled intervention study using etanercept (a monoclonal antibody which works as a TNFα antagonist) was carried out. Etanercept treatment at GD0, 6, 12 and 18 resulted in an improvement in pregnancy outcome in the SHRSP with an increased litter size and reduced resorption rate. Furthermore, there was an improved uterine artery function in GD18 SHRSP treated with etanercept which was associated with an improved uterine artery blood flow over the course of gestation. In Chapter 3, I sought to identify the source of this detrimental excess of TNFα by designing a panel for maternal leukocytes in the blood and placenta at GD18. A population of CD3- CD161+ cells, which are defined as rat natural killer (NK) cells, were increased in number in the SHRSP. Intracellular flow cytometry also identified this cell type as a source of excess TNFα in blood and placenta from pregnant SHRSP. I then went on to evaluate the effects of etanercept treatment on these CD3- CD161+ cells and showed that etanercept reduced the expression of CD161 and the cytotoxic molecule, granzyme B, in the NK cells. Thus, etanercept limits the cytotoxicity and potential damaging effect of these NK cells in the SHRSP placenta. Analysing the urinary peptidome has clinical potential to identify novel pathways involved with disease and/or to develop biomarker panels to aid and stratify diagnosis. In Chapter 4, I utilised the SHRSP as a pre-clinical model to identify novel urinary peptides associated with hypertensive pregnancy. Firstly, a characterisation study was carried out in the kidney of the WKY and SHRSP. Urine samples from WKY and SHRSP taken at pre-pregnancy, mid-pregnancy (GD12) and late pregnancy (GD18) were used in the peptidomic screen. In order to capture peptides which were markers of hypertensive pregnancy from the urinary peptidomic data, I focussed on those that were only changed in a strain dependent manner at GD12 and 18 and not pre-pregnancy. Peptide fragments from the uromodulin protein were identified from this analysis to be increased in pregnant SHRSP relative to pregnant WKY. This increase in uromodulin was validated at the SHRSP kidney level using qPCR. Uromodulin has previously been identified to be a candidate molecule involved in systemic arterial hypertension but not in hypertensive pregnancy thus is a promising target for further study. In summary, we have characterised the SHRSP as the first model of maternal chronic hypertension during pregnancy and identified that inflammation mediated by TNFα and NK cells plays a key role in the pathology. The evidence presented in this thesis establishes the SHRSP as a pre-clinical model for pregnancy research and can be continued into clinical studies in pregnant women with chronic hypertension which remains an area of unmet research need.

First reproductive and productive results on “Ribatejano pig”

The present trail aimed to study the effect of crossbreeding between Alentejano (AL) and Bísaro (BI) swine breeds (“Ribatejano pig”) on some reproductive and productive traits. Nine AL gilts and sows and six BI gilts were crossed with BI and AL boars, respectively. Mating and farrowing dates, prolificacy and litter size at 28d were registered for all sows. The pregnancy length of AL sows was shorter (111±0.4d vs 113.7±0.5d; p=0.002) than the observed on BI females. The BI gilts presented higher prolificacy rate than AL on both total born (11.0±1.0 vs 6.7±0.8; P=0.004) and born alive piglets (10.0±1.0 vs 6.7±0.8; p=0.026). The mortality rate was similar in both genotypes (p=0.255) being on average 12%, therefore at 28d after farrowing the litter size remained higher in BI sows (8.5±0.8 vs 6.1±0.6; p= 0.032). A subset of each genotype (4 gilts) was supervised during farrowing and lactation (until 28d) and piglets were weighed at birth, 24h and 28d of live. Farrowing length was not significantly different (p=0.253) between genotypes, averaging 97±22 min. When compared to ALBI (AL x BI) piglets, the BIAL (BI x AL) piglets were heavier at birth (1402±46g vs 1209±36g; p=0.002). Colostrum intake of piglets per kg of birth weight on the first 24h of life was similar between genotypes (p=0.735) being 289±15g for ALBI and 281±19g for BIAL piglets. The growth rate of piglets from birth to 28d and piglet weight at 28d was not different between genotypes (p=0.161 and p=0.091) averaging 195±6g and 6761±181g, respectively. Litter weight at 28d tended (P=0.06) to be higher on ALBI litters (56.6±4.0kg) than BIAL litters (43.2±4.0kg). This results obtained within the frame of Treasure project* are, at our knowledge, the first data of these crossbred piglets and could be used in future as reference for further studies and also for farmers that may try these cross on a commercial basis.

Climate seasonality limits leaf carbon assimilation and wood productivity in tropical forests.

The seasonal climate drivers of the carbon cy- cle in tropical forests remain poorly known, although these forests account for more carbon assimilation and storage than any other terrestrial ecosystem. Based on a unique combina- tion of seasonal pan-tropical data sets from 89 experimental sites (68 include aboveground wood productivity measure- ments and 35 litter productivity measurements), their asso- ciated canopy photosynthetic capacity (enhanced vegetation index, EVI) and climate, we ask how carbon assimilation and aboveground allocation are related to climate seasonal- ity in tropical forests and how they interact in the seasonal carbon cycle. We found that canopy photosynthetic capacity seasonality responds positively to precipitation when rain- fall is < 2000 mm yr-1 (water-limited forests) and to radia- tion otherwise (light-limited forests). On the other hand, in- dependent of climate limitations, wood productivity and lit- terfall are driven by seasonal variation in precipitation and evapotranspiration, respectively. Consequently, light-limited forests present an asynchronism between canopy photosyn- thetic capacity and wood productivity. First-order control by precipitation likely indicates a decrease in tropical forest pro- ductivity in a drier climate in water-limited forest, and in cur- rent light-limited forest with future rainfall < 2000 mm yr-1.

Decomposition of organic matter in the presence of the herbicide clomazone.

Clomazone (2-(2-chlorophenyl) methyl-4.4-dimethyl-3-isoxazolidinone) is a post emergence herbicide widely used in rice fields in Rio Grande do Sul (Brazil) with high activity against Gramineae at the recommended application rate of 700 g/ha. The presence of this chemical in the water may affect microorganisms responsible for the decomposition of organic matter. Thus, a disturbe in the trophic chain sustained by the decompositors could happen. In the present work the decomposition rate of organic matter (Typha latifolia) exposed to several concentrations of a clomazone formulation: 0 (control), 25.0, 62.0, 156.0, 390.0 and 976.0mg/L on the basis of the active ingredient was evaluated. Five litter bags containing about 3.0g of pieces of T. latifolia leaves wereplaced in aquariums with 15 of reconstituted water. In cach aquarium were added 500g of sediment from the same place of the plant collection, as a source of decompositors microorganisms. The results relative tothe control, showed that the decomposition rate in the highest and lowest dose was reduced in 50.05 and 1,28%, respectively, after 80 days.

Produção e degradação da serrapilheira de espécies lenhosas da caatinga e sua contribuição na recuperação de áreas degradadas.

Resumo: O entendimento do fluxo de produção e do aporte de nutrientes via decomposição da serrapilheira e as interações do processo com parâmetros edáficos e ciclagem de nutrientes de espécies nativas da Caatinga têm sido pouco estudados. O conhecimento sobre ciclagem de nutrientes em florestas manejadas também permite inferências sobre as espécies com maior capacidade de reciclagem de nutrientes e seu potencial para recuperação de áreas degradadas. Objetivou-se com isso avaliar a produção e a degradação da serrapilheira de oito espécies lenhosas da Caatinga e mensurar os efeitos de sua aplicação sobre a fertilidade do solo e sobre a produção de sorgo em solo degradado. Para isso realizou-se três ensaios: para o ensaio I quantificou-se a produção de serrapilheira em um delineamento inteiramente casualizado com 6 repetições, por meio da instalação de coletores sob a projeção da copa das espécies (tratamentos): mofumbo, sabiá, jurema-preta, jucá, catingueira, pereiro, pau-branco e marmeleiro, sendo o material coletado mensalmente; foram quantificadas a produção das frações folhas, caule, material reprodutivo, miscelânea e total, bem como o aporte de nutrientes no período chuvoso e seco. Para o ensaio II avaliou-se a taxa de degradação da fração folhas de cada espécie citada por meio da utilização de litter bags, em delineamento inteiramente casualizado com 4 repetições, as coletas foram aos 0, 30, 60, 90, 120 e 150 dias, em seguida quantificou-se os macro e micronutrientes, celulose, lignina e carbono em cada tempo de amostragem. Para o ensaio III, realizou-se experimento em casa de vegetação para mensurar os efeitos da aplicação dos resíduos da serrapilheira das mesmas espécies mencionadas nos ensaios anteriores (I e II) sobre a fertilidade do solo e a produção de sorgo em solo degradado, neste experimento adotou-se o delineamento em blocos casualizados com 5 tratamentos e 5 repetições, sendo avaliadas doses equivalentes a: 0, 15, 30, 60 e 120 kg ha-1 de N dos resíduos de cada espécie e um tratamento adicional com adubação mineral, totalizando 30 unidades experimentais para cada espécie. As variáveis mensuradas foram biométricas, biomassa, teor relativo de clorofila e nitrogênio total, além de análises de fertilidade do solo. Com a análise dos dados verificou-se que a época de maior produção de serrapilheira ocorreu no final do período chuvoso para o início do período seco. A espécie jucá apresentou maior produção de serrapilheira, comparado às outras espécies. O nutriente cálcio apresentou maior acúmulo na serrapilheira para as espécies mofumbo, sabiá, catingueira, pereiro e marmeleiro e o nitrogênio foi superior para as espécies jurema-preta, jucá e pau-branco. Para todas as espécies avaliadas no ensaio de degradação houve redução significativa na sua biomassa em relação ao tempo zero, apresentando a seguinte ordem de velocidade de decomposição: jurema-preta > catingueira > pau-branco > jucá > marmeleiro > mofumbo > pereiro > sabiá. No ensaio de fertilização com os resíduos verificou-se que o marmeleiro promoveu efeitos negativos no solo, como acidificação. Porém, a aplicação dos resíduos da espécie pau-branco foi a que promoveu aumento nos valores de K, SB e CEC do solo e na produção do sorgo os resíduos de jurema-preta e pau-branco foram as que promoveram aumento na massa seca das plantas. Enquanto a adubação mineral proporcionou aumento na produção de massa seca do sorgo, demonstrando que a associação entre adubo mineral e o uso da serrapilheira de espécies da Caatinga pode ser uma opção viável para acelerar a recuperação de solos degradados. Abstract: The understanding of the production flow and nutrient supply via decomposition of litter and process interactions with edaphic parameters and nutrient cycling of native species of the Caatinga has been little studied. The knowledge of nutrient cycling in managed forests also allow inferences about species with capacity greater nutrient recycling capacity and its potential for recovery of degraded areas. This study aimed to evaluate the production and litter degradation 8 woody species of Caatinga and measure the effects of its application on soil fertility and production of sorghum in degraded soil. To this was carried out three tests: for the test I quantified the production of litter in a completely randomized design with 6 replications, by installing collectors under the canopy projection in the species (treatments): mofumbo, sabiá, jurema-preta, jucá, catingueira, pereiro, pau-branco and marmeleiro for each species, and the material collected monthly, were quantified the production of fractions leaves, stem, reproductive material, miscellany and total nutrient intake in the rainy and dry season. For II test evaluated the degradation rate of the fraction leaves through the use of litter bags, in a completely randomized design with 4 replications, the collected was 0, 30, 60, 90, 120 and 150 days and quantitated nutrients, cellulose, lignin and carbon at each evaluation time. For the III test, there was the experiment in a greenhouse to measure the effects of the application of litter waste of the same species of previous tests (I and II) on soil fertility and production of sorghum in degraded soil, was adopted the randomized block design with 5 treatments and 5 replications and evaluated doses equivalent to: 0, 15, 30, 60 and 120 kg ha-1 N of waste each species and an additional treatment with mineral fertilizer, totaling 30 experimental units for each species. Biometric analysis and biomass, relative chlorophyll content and total nitrogen were proceeded. In addition to soil fertility analysis. With the data analysis it was found that the time of greatest litterfall occurred at the end of the rainy season to the beginning of the dry season. The jucá species showed higher production compared to other species. The nutrient calcium had higher accumulation for the species mofumbo, sabiá, catingueira, pereiro and marmeleiro and nitrogen was higher for species jurema-preta, jucá and pau-branco. All species evaluated in degradation test had a significant reduction in biomass over time zero. They presented the following order of decomposition rate: jurema-preta > catingueira > pau-branco > jucá > marmeleiro > mofumbo > pereiro > sabiá. For fertility test it was found that marmeleiro promoted negative effects on soil, such as acidification. However, pau-branco was the specie that promoted further improvements in the K values, SB and CEC to the soil and for the production of sorghum, the waste jurema-preta and pau-branco promoted increase in dry matter plants. While the mineral fertilization provided an increase in dry matter production of sorghum, demonstrating that the combination of mineral fertilizer and the use of litter of Caatinga species may be a viable option to speed up the recovery of degraded soils.