Trace elements and small-usage compounds can be transported from Earth, or in some cases extracted from the Martian regolith. In the case where power is provided from photocollection or photovoltaics, light energy will vary with location and season, and may be critical to power our bioreactors. Although photosynthetic organisms are attractive for FPS, a higher demand for carbon-rich feedstocks and other chemicals necessitates a more rapid and efficient CO2 fixation strategy. Physicochemical conversion is inefficient due to high temperature and pressure requirements. Microbial electrosynthesis , whereby reducing power is passed from abiotic electrodes to microbes to power CO2 reduction, can offer rapid and efficient CO2 fixation at ambient temperature and pressure . MES can produce a variety of chemicals including acetate , isobutanol , PHB , and sucrose , and therefore represents a filexible and highly promising ISRU platform technology . Biological N2-fixation offers power- and resource-efficient ammonium production. Although photoautotrophic N2 fixation with, for example, purple non-sulfur bacteria, is possible, slow growth rates due to the high energetic demand of nitrogenase limit throughput . Therefore, heterotrophic production with similar bacteria using acetate or sucrose as a feed stock sourced from electromicrobial CO2-fixation represents the most promising production scheme, and additionally benefits from a high degree of process redundancy with heterotrophic bioplastic production. Regolith provides a significant inventory for trace elements and, when mixed with the substantial cellulosic biomass waste from FPS processes, can facilitate recycling organic matter into fertilizer to support crop growth. However, regolith use is hampered by widespread perchlorate , indicating that decontamination is necessary prior to enrichment or use. Dechlorination can be achieved via biological perchlorate reduction using one of many dissimilatory perchlorate reducing organisms . Efforts to reduce perchlorate biologically have been explored independently and in combination with a more wholistic biological platform . Such efforts to integrate synthetic biology into human exploration missions suggest that a number of approaches should be considered within a surface bio-manufactory.

A biomanufactory must be able to produce and utilize feed stocks along three axes as depicted in Figure 5: CO2-fixation to supply a carbon and energy source for downstream heterotrophic organisms or to generate commodity chemicals directly, N2-fixation to provide ammonium and nitrate for plants and non-diazotrophic microbes,macetas para viveros and regolith decontamination and enrichment for soil-based agriculture and trace nutrient provision. ISRU inputs are sub-module and organism dependent, with all sub-modules requiring water and power. For the carbon fixation sub-module , CO2 is supplied as the carbon source, and electrons are supplied as H2 or directly via a cathode. Our proposed bio-catalysts are the lithoautotrophic Cupriavidus necator for longer-chain carbon production [e.g., sucrose ] and the acetogen Sporomusa ovata for acetate production. C. necator is a promising chassis for metabolic engineering and scale-up , with S. ovata having one of the highest current consumptions for acetogens characterized to date . The fixed-carbon outputs of this sub-module are then used as inputs for the other ISRU sub-modules in addition to the ISM module . The inputs to the nitrogen fixation sub-module include fixed carbon feed stocks, N2, and light. The diazotrophic purple-non sulfur bacterium Rhodopseudomonas palustris is the proposed bio-catalyst, as this bacterium is capable of anaerobic, light-driven N2 fixation utilizing acetate as the carbon source, and has a robust genetic system allowing for rapid manipulation . The output product is fixed nitrogen inthe form of ammonium, which is used as a feed stock for the carbon-fixation sub-module of ISRU along with the FPS and ISM modules. The inputs for the regolith enrichment sub-module include regolith, fixed carbon feedstocks, and N2. Azospira suillum is a possible bio-catalyst of choice due to its dual use in perchlorate reduction and nitrogen fixation . Regolith enrichment outputs include soil for the FPS module , H2 that can be fed back into the carbon fixation sub-module and the ISM module, chlorine gas from perchlorate reduction, and waste products. Replicate ISRU bioreactors operating continuously in parallel with back-up operations lines can ensure a constant supply of the chemical feed stocks, commodity chemicals, and biomass for downstream processing in ISM and FPS operations. Integration of ISRU technologies with other biomanufactory elements, especially anaerobic digestion reactors, may enable complete recyclability of raw materials, minimizing resource consumption and impact on the Martian environment .

Waste stream processing to recycle essential elements will reduce material requirements in the biomanufactory. Typical feed stocks include inedible crop mass, human excreta, and other mission wastes. Space mission waste management traditionally focuses on water recovery and efficient waste storage through warm air drying and lyophilization . Mission trash can be incinerated to produce CO2, CO, and H2O . Pyrolysis, another abiotic technique, yields CO and H2 alongside CH4 . The Sabatier process converts CO2 and CO to CH4 by reacting with H2. An alternate thermal degradation reactor , operating under varying conditions that promote pyrolysis, gasification, or incineration, yields various liquid and gaseous products. The fact remains however, that abiotic carbon recycling is inefficient with respect to desired product CH4, and is highly energy-intensive. Microbes that recover resources from mission wastes are a viable option to facilitate loop closure. Aerobic composting produces CO2 and a nutrient-rich extract for plant and microbial growth . However, this process requires O2, which will likely be a limited resource. Hence, anaerobic digestion, a multi-step microbial process that can produce a suite of endproducts at lower temperature than abiotic techniques , is the most promising approach for a Mars biomanufactory to recycle streams for the ISM and FPS processes. Digestion products CH4 and volatile fatty acids can be substrates for polymer-producing microbes . Digestate, with nutrients of N, P, and K, can be ideal for plant and microbial growth , as shown in Figure 6. Additionally, a CH4 and CO2 mixture serves as a biogas energy source, and byproduct H2 is also an energy source . Because additional infrastructure and utilities are necessary for waste processing, the extent of loop closure that is obtainable from a treatment route must be analyzed to balance yield with its infrastructure and logistic costs. Anaerobic digestion performance is a function of the composition and pretreatment of input waste streams , as well as reaction strategies like batch or continuous, number of stages, and operation conditions such as organic loading rate, solids retention time, operating temperature, pH, toxic levels of inhibitors and trace metal requirements . Many of these process parameters exhibit trade-offs between product yield and necessary resources. For example, a higher waste loading reduces water demand, albeit at the cost of process efficiency. There is also a potential for multiple co-benefits of anaerobic digestion within the biomanufactory. Anaerobic biodegradation of nitrogen-rich protein feed stocks, for example, releases free NH3 by ammonification. While NH3 is toxic to anaerobic digestion and must thus be managed , it reacts with carbonic acid to produce bicarbonate buffer and ammonium, decreasing CO2 levels in the biogas and buffering against low pH.

The resulting digestate ammonium can serve as a fertilizer for crops and nutrient for microbial cultures.FPS and ISM waste as well as human waste are inputs for an anaerobic digester, with output recycled products supplementing the ISRU unit. Depending on the configuration of the waste streams from the biomanufactory and other mission elements, the operating conditions of the process can be varied to alter the efficiency and output profile. Open problems include the design and optimization of waste processing configurations and operations, and the identification of optimal end-product distributions based on a loop closure metric against mission production profiles, mission horizon, biomanufacturing feedstock needs, and the possible use of leftover products by other mission elements beyond the bio-manufactory. A comparison with abiotic waste treatment strategies is also needed, checking power demand, risk, autonomy, and modularity benefits.Biomanufactory development must be done in concert with planned NASA missions that can provide critical opportunities to test subsystems and models necessary to evaluate efficacy and technology readiness levels . Figure 7 is our attempt to place critical elements of a biomanufactory road map into this context. We label critical mission stages using Reference Mission Architecture -S and RMA-L,macetas por mayor which refer to Mars surface missions with short and long durations, respectively.Beyond contamination, there are ethical issues that concern both the act of colonizing a new land and justifying the cost and benefits of a mission given needs of the many here on earth. Our road map begins with the call for an extensive and ongoing discussion of ethics . Planetary protection policies can provide answers or frameworks to address extant ethical questions surrounding deep-space exploration, especially on Mars . Critically, scientists and engineers developing these technologies cannot be separate or immune to such policy development.We have outlined the design and future deployment of a biomanufactory to support human surface operations during a 500 days manned Mars mission. We extended previous stand-alone biological elements with space use potential into an integrated biomanufacturing system by bringing together the important systems of ISRU, synthesis, and recycling, to yield food, pharmaceuticals, and bio-materials. We also provided an envelope of future design, testing, and biomanufactory element deployment in a road map that spans Earth-based system development, testing on the ISS, integration with lunar missions, and initial construction during shorter-term initial human forays on Mars. The innovations necessary to meet the challenges of low-cost, energy and mass efficient, closed-loop, and regenerable bio-manufacturing for space will undoubtedly yield important contributions to forwarding sustainable bio-manufacturing on Earth. We anticipate that the path towards instantiating a biomanufactory will be replete with science, engineering, and ethical challenges. But that is the excitement—part-and-parcel—of the journey to Mars.Rose production is currently the largest component of California’s $300 million cut-flower industry. In 2001, California growers produced 66% of the U.S. rose crop, with a wholesale value of $45 million . The key pests of cut roses are two spotted spider mites , western flower thrips and rose powdery mildew . The two spotted spider mite is a foliage feeder that extracts the cell contents from leaves. This feeding causes foliar stippling and can disrupt the plant’s photosynthetic and water balance mechanisms . The western flower thrips is both a foliage and flower feeder, although it feeds primarily on flowers in the cut-rose system . Powdery mildew is probably the most widespread and best-known disease of roses. The fungus produces a white, powdery-appearing growth of mycelium and conidia on leaves, which can cause distortion, discoloration and premature senescence. Although it causes some disruption of photosynthesis and transpiration control, the key impact of powdery mildew is reduced aesthetic value caused by the white, powdery spots and leaf distortion. Fresh cut roses are often harvested twice daily, so revised reentry intervals imposed by the U.S. Environmental Protection Agency after pesticide application limit the number of pesticides that are useful in this production system . In addition, the typical number of pesticide sprays applied to roses grown for cut flowers has impeded the implementation of integrated pest management procedures, particularly the use of biological controls. The IPM approach to pest management incorporates all cost-effective control tactics appropriate for the crop, including biological, cultural and chemical controls. Pesticides that target hard-to-kill floriculture pests frequently kill natural enemies as well, which favors continued reliance on conventional pesticides while discouraging the adoption of biological control. Heavy pesticide use against key pests in the greenhouse has resulted in the widespread development of pesticide resistance in western flower thrips , mites , white flies , aphids and leaf miners . The heavy use of pesticides in cut roses is also a worker safety concern in global and local production. California rose growers reached a crisis point about 8 years ago, when pesticide resistance, costs and limited pesticide availability threatened the growers’ ability to effectively manage two spotted spider mites. At the same time, a new cut-rose production system that favors the success of IPM was gaining widespread acceptance. Roses were traditionally grown in soil with a hedgerow training system, where flowers are cut in a manner that gradually creates a 7-foot or taller hedge. The hedges are pruned back annually to about a 3-foot height and the process is begun again.