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Kentucky Annual Economic Report 2023, Michael Childress, Michael W. Clark, James P. Ziliak, Simon Sheather

Kentucky Annual Economic Report

This report is one of the important ways that the Center for Business and Economic Research fulfills its mission to examine various aspects of Kentucky’s economy as directed by the Kentucky Revised Statutes (KRS 164.738). The analysis and data presented here cover a variety of topics that range from a discussion of Kentucky’s current economic climate to a broad presentation of factors affecting the economy.

Beliefs And Knowledge Related To Human Papillomavirus (Hpv) Vaccine Among African Americans And African Immigrants Young Adults, Adebola Adegboyega, Omoadoni Roseline Obielodan, Amanda T. Wiggins, Mark Dignan, Lovoria B. Williams

UK CARES Faculty Publications

Background Despite the disparate human papillomavirus (HPV) infection rates among sexually active Black young adults, HPV vaccine uptake remains low among this population. This study aimed to explore HPV beliefs, attitudes, and knowledge among Black young adults and provide recommendations on ways to improve vaccine uptake.

Methods We used a mixed-method, convergent design to conduct five focus groups and administered a 40-item electronic survey that was developed with health belief model (HBM) constructs. We assessed HPV and vaccine knowledge, barriers, and attitudes toward vaccination. We analyzed quantitative data using descriptive statistics and bivariate methods. Focus group transcripts were analyzed using …

Higher Concentrations Of Microplastics In Runoff From Biosolid-Amended Croplands Than Manure-Amended Croplands, Nasrin Naderi Beni, Shahab Karimifard, John Gilley, Tiffany L. Messer, Amy Schmidt, Shannon Bartelt-Hunt

UK CARES Faculty Publications

Land-applied municipal biosolids, produced from municipal wastewater treatment sludge, contributes to microplastics contamination in agroecosystems. The impacts of biosolids on microplastic concentrations in agricultural soil have been previously investigated, however, the potential for microplastics transport from biosolid-amended croplands has not been previously quantified. In this study, manure and biosolids were applied to field plots, runoff was collected following natural precipitation events and the potential of bacterial biofilm to grow on different microplastic morphologies was investigated. Higher concentrations of microplastics were detected in runoff from plots with land-applied biosolid in comparison with manure-amended and control plots. Fibers and fragments were the …

Forage News [2023-02], Department Of Plant And Soil Sciences, University Of Kentucky

Forage News

No abstract provided.

Novel Smad3 Variant Identified In A Patient With Familial Aortopathy Modeled Using A Zebrafish Embryo Assay, Mary Burchett Sheppard, Jeffrey D. Smith, Lisa L. Bergmann, Jakub Konrad Famulski

Saha Cardiovascular Research Center Faculty Publications

In human, pathogenic variants in smad3 are one cause of familial aortopathy. We describe a novel SMAD3 variant of unknown significance (VUS), V244F, in a patient who presented with aortic root dilation, right coronary artery ectasia, abdominal aortic aneurysm, right vertebral artery atresia, and cavernoma. Determination of variant pathogenicity impacted multiple aspects of the patient’s care, including the most appropriate surgical threshold for which to recommend a valve-sparing aortic root replacement. To determine whether the newly identified SMAD3 variant, and whether SMAD3 induced aortopathy in general, can be assayed in a zebrafish embryo model, we injected smad3a mRNA into Tg[kdrl:mCherry] …

Process Intensification Strategies For Green Solvent Mediated Biomass Pretreatment, Ahamed Ullah, Yuxuan Zhang, Can Liu, Qi Qiao, Qing Shao, Jian Shi

Markey Cancer Center Faculty Publications

Demonstrated to be highly effective for lignocellulosic biomass pretreatment, deep eutectic solvent (DES) has attracted increasing attention owing to its advantages of simple synthesis, relatively low chemical cost, and better biocompatibility as compared to certain ionic liquids. Here we provide a critical review of the status of the design/selection of DES for the pretreatment of biomass feedstocks with an emphasis on the process intensification strategies: 1) integration of microwave, ultrasound, and high solid extrusion for pretreating biomass, 2) one-pot DES pretreatment, enzymatic hydrolysis, and fermentation, 3) strategies for DES recycling and product recovery; and 4) recent progress on molecular simulations …

Aortic Stress Activates An Adaptive Program In Thoracic Aortic Smooth Muscle Cells That Maintains Aortic Strength And Protects Against Aneurysm And Dissection In Mice, Chen Zhang, Yanming Li, Abhijit Chakraborty, Yang Li, Kimberly R. Rebello, Pingping Ren, Wei Luo, Lin Zhang, Hong S. Lu, Lisa A. Cassis, Joseph S. Coselli, Alan Daugherty, Scott A. Lemaire, Ying H. Shen

Saha Cardiovascular Research Center Faculty Publications

BACKGROUND: When aortic cells are under stress, such as increased hemodynamic pressure, they adapt to the environment by modifying their functions, allowing the aorta to maintain its strength. To understand the regulation of this adaptive response, we examined transcriptomic and epigenomic programs in aortic smooth muscle cells (SMCs) during the adaptive response to AngII (angiotensin II) infusion and determined its importance in protecting against aortic aneurysm and dissection (AAD).

METHODS: We performed single-cell RNA sequencing and single-cell sequencing assay for transposase-accessible chromatin (scATAC-seq) analyses in a mouse model of sporadic AAD induced by AngII infusion. We also examined the direct …

2022 Kentucky River Watershed Watch Annual Report, Steven J. Evans, Malissa Mcalister

Kentucky River Watershed Watch

No abstract provided.

Tris(1,3-Dichloro-2-Propyl) Phosphate Is A Metabolism-Disrupting Chemical In Male Mice, Sara Y. Ngo Tenlep, Megan L. Weaver, Jianzhong Chen, Olga A. Vsevolozhskaya, Andrew J. Morris, Cetewayo S. Rashid

Pharmacology and Nutritional Sciences Faculty Publications

Tris(1,3-dichloro-2-propyl) phosphate (TDCPP) is an organophosphate flame retardant. The primary TDCPP metabolite, bis(1,3-dichloro-2-propyl) phosphate (BDCPP), is detectable in the urine of over 90 % of Americans. Epidemiological studies show sex-specific associations between urinary BDCPP levels and metabolic syndrome, which is an established risk factor for type 2 diabetes, heart disease, and stroke. We used a mouse model to determine whether TDCPP exposure disrupts glucose homeostasis. Six-week old male and female C57BL/6J mice were given ad libitum access to diets containing vehicle (0.1 % DMSO) and TDCPP resulting in the following treatment groups: 0 mg/kg/day, 0.02 mg/kg/day, 1 mg/kg/day, or 100 …

Gasdermin D Deficiency In Vascular Smooth Muscle Cells Ameliorates Abdominal Aortic Aneurysm Through Reducing Putrescine Synthesis, Jianing Gao, Yanghui Chen, Huiqing Wang, Xin Li, Ke Li, Yangkai Xu, Xianwei Xie, Yansong Guo, Nana Yang, Xinhua Zhang, Dong Ma, Hong S. Lu, Ying H. Shen, Yong Liu, Jifeng Zhang, Y. Eugene Chen, Alan Daugherty, Dao Wen Wang, Lemin Zheng

Saha Cardiovascular Research Center Faculty Publications

Abdominal aortic aneurysm (AAA) is a common vascular disease associated with significant phenotypic alterations in vascular smooth muscle cells (VSMCs). Gasdermin D (GSDMD) is a pore-forming effector of pyroptosis. In this study, the role of VSMC-specific GSDMD in the phenotypic alteration of VSMCs and AAA formation is determined. Single-cell transcriptome analyses reveal Gsdmd upregulation in aortic VSMCs in angiotensin (Ang) II-induced AAA. VSMC-specific Gsdmd deletion ameliorates Ang II-induced AAA in apolipoprotein E (ApoE)−/− mice. Using untargeted metabolomic analysis, it is found that putrescine is significantly reduced in the plasma and aortic tissues of VSMC-specific GSDMD deficient mice. High putrescine levels …

The Mechanism And Therapy Of Aortic Aneurysms, Jianing Gao, Huanhuan Cao, Gaofei Hu, Yufei Wu, Yangkai Xu, Hongtu Cui, Hong S. Lu, Lemin Zheng

Saha Cardiovascular Research Center Faculty Publications

Aortic aneurysm is a chronic aortic disease affected by many factors. Although it is generally asymptomatic, it poses a significant threat to human life due to a high risk of rupture. Because of its strong concealment, it is difficult to diagnose the disease in the early stage. At present, there are no effective drugs for the treatment of aneurysms. Surgical intervention and endovascular treatment are the only therapies. Although current studies have discovered that inflammatory responses as well as the production and activation of various proteases promote aortic aneurysm, the specific mechanisms remain unclear. Researchers are further exploring the pathogenesis …

Abl1/2 And Ddr1 Drive Meki Resistance In Nras-Mutant Melanomas By Stabilizing Raf/Myc/Ets1 And Promoting Raf Homodimerization, Anastasia Lyon, Rakshamani Tripathi, Christina Meeks, Daheng He, Yuanyuan Wu, Jinpeng Liu, Chi Wang, Jing Chen, Haining Zhu, Sujata Mukherjee, Saptadwipa Ganguly, Rina Plattner

Markey Cancer Center Faculty Publications

Melanomas harboring NRAS mutations are a particularly aggressive and deadly subtype. If patients cannot tolerate or the melanomas are insensitive to immune checkpoint blockade, there are no effective 2nd-line treatment options. Drugs targeting the RAF/MEK/ERK pathway, which are used for BRAF-mutant melanomas, do little to increase progression-free survival (PFS). Here, using both loss-of-function and gain-of-function approaches, we show that ABL1/2 and DDR1 are critical nodes during NRAS-mutant melanoma intrinsic and acquired MEK inhibitor (MEKi) resistance. In some acquired resistance cells, ABL1/2 and DDR1 cooperate to stabilize RAF proteins, activate ERK cytoplasmic and nuclear signaling, repress p27/KIP1 expression, and drive RAF …

Adolescent- And Adult-Initiated Alcohol Exposure In Mice Differentially Promotes Tumorigenesis And Metastasis Of Breast Cancer, Mei Xu, Hui Li, Danlei Chen, Huaxun Wu, Wen Wen, Hong Xu, Jacqueline Frank, Gang Chen, Jia Luo

Markey Cancer Center Faculty Publications

Background: Alcohol exposure increases the risk of breast cancer. Alcohol consumption by adolescents is a serious social and public health issue. This study investigated the impact of adolescent alcohol consumption on mammary tumorigenesis and progression and compared it to that of adult alcohol exposure in animal models. Methods: Female adolescent (5 weeks) and adult (8 weeks) MMTV-Wnt1 mice were exposed to alcohol either chronically or acutely. For chronic alcohol exposure, animals were fed a liquid diet containing 6.7% ethanol for 23 weeks. For acute exposure, animals were treated with ethanol (2.5 g/kg, 25% w/v) via intraperitoneal (IP) injection for 15 …

Bilirubin Nanoparticle Treatment In Obese Mice Inhibits Hepatic Ceramide Production And Remodels Liver Fat Content, Zachary A. Kipp, Genesee J. Martinez, Evelyn A. Bates, Agil B. Maharramov, Robert M. Flight, Hunter N. B. Moseley, Andrew J. Morris, David E. Stec, Terry D. Hinds Jr.

Markey Cancer Center Faculty Publications

Studies have indicated that increasing plasma bilirubin levels might be useful for preventing and treating hepatic lipid accumulation that occurs with metabolic diseases such as obesity and diabetes. We have previously demonstrated that mice with hyperbilirubinemia had significantly less lipid accumulation in a diet-induced non-alcoholic fatty liver disease (NAFLD) model. However, bilirubin’s effects on individual lipid species are currently unknown. Therefore, we used liquid chromatography-mass spectroscopy (LC-MS) to determine the hepatic lipid composition of obese mice with NAFLD treated with bilirubin nanoparticles or vehicle control. We placed the mice on a high-fat diet (HFD) for 24 weeks and then treated …

Clic And Membrane Wound Repair Pathways Enable Pandemic Norovirus Entry And Infection, B. Vijayalakshmi Ayyar, Khalil Ettayebi, Wilhelm Salmen, Umesh C. Karandikar, Frederick H. Neill, Victoria R. Tenge, Sue E. Crawford, Erhard Bieberich, B. V. Venkataram Prasad, Robert L. Atmar, Mary K. Estes

Markey Cancer Center Faculty Publications

Globally, most cases of gastroenteritis are caused by pandemic GII.4 human norovirus (HuNoV) strains with no approved therapies or vaccines available. The cellular pathways that these strains exploit for cell entry and internalization are unknown. Here, using nontransformed human jejunal enteroids (HIEs) that recapitulate the physiology of the gastrointestinal tract, we show that infectious GII.4 virions and virus-like particles are endocytosed using a unique combination of endosomal acidification-dependent clathrin-independent carriers (CLIC), acid sphingomyelinase (ASM)-mediated lysosomal exocytosis, and membrane wound repair pathways. We found that besides the known interaction of the viral capsid Protruding (P) domain with host glycans, the Shell …

Contouring Quality Assurance Methodology Based On Multiple Geometric Features Against Deep Learning Auto-Segmentation, Jingwei Duan, Mark E. Bernard, James Castle, Xue Feng, Chi Wang, Mark C. Kenamond, Quan Chen

Markey Cancer Center Faculty Publications

Background: Contouring error is one of the top failure modes in radiation treatment. Multiple efforts have been made to develop tools to automatically detect segmentation errors.Deep learning-based auto-segmentation (DLAS) has been used as a baseline for flagging manual segmentation errors, but those efforts are limited to using only one or two contour comparison metrics.

Purpose: The purpose of this research is to develop an improved contouring quality assurance system to identify and flag manual contouring errors.

Methods and materials: DLAS contours were used as a reference to compare with manually segmented contours. A total of 27 geometric agreement metrics were …

Critical Role Of The Sulfiredoxin-Peroxiredoxin Iv Axis In Urethane-Induced Non-Small Cell Lung Cancer, Yanning Hao, Hong Jiang, Pratik Thapa, Na Ding, Aziza Alshahrani, Junichi Fujii, Michel B. Toledano, Qiou Wei

Markey Cancer Center Faculty Publications

Non-small cell lung cancer (NSCLC), the most common type of lung cancer, etiologically associates with tobacco smoking which mechanistically contributes to oxidative stress to facilitate the occurrence of mutations, oncogenic transformation and aberrantly activated signaling pathways. Our previous reports suggested an essential role of Sulfiredoxin (Srx) in promoting the development of lung cancer in humans, and was causally related to Peroxiredoxin IV (Prx4), the major downstream substrate and mediator of Srx-enhanced signaling. To further explore the role of the Srx-Prx4 axis in de novo lung tumorigenesis, we established Prx4−/− and Srx−/−/Prx4−/− mice in pure FVB/N background. Together with wild-type litter …

Ifn-Γ Production By Brain-Resident Cells Activates Cerebral Mrna Expression Of A Wide Spectrum Of Molecules Critical For Both Innate And T Cell-Mediated Protective Immunity To Control Reactivation Of Chronic Infection With Toxoplasma Gondii, Yasuhiro Suzuki, Jenny Lutshumba, Kuey Chu Chen, Mohamed H. Abdelaziz, Qila Sa, Eri Ochiai

Markey Cancer Center Faculty Publications

We previously demonstrated that brain-resident cells produce IFN-g in response to reactivation of cerebral infection with Toxoplasma gondii. To obtain an overall landscape view of the effects of IFN-g from brain-resident cells on the cerebral protective immunity, in the present study we employed NanoString nCounter assay and quantified mRNA levels for 734 genes in myeloid immunity in the brains of T and B cell-deficient, bone marrow chimeric mice with and without IFN-g production by brain-resident cells in response to reactivation of cerebral T. gondii infection. Our study revealed that IFN-g produced by brain-resident cells amplified mRNA expression for the molecules …

Lignin Derived Hydrophobic Deep Eutectic Solvents As Sustainable Extractants, Yuxuan Zhang, Qi Qiao, Usman Lame Abbas, Jun Liu, Yi Zheng, Christopher Jones, Qing Shao, Jian Shi

Markey Cancer Center Faculty Publications

Solvent innovation has become a central task for improving the sustainability of chemical processes1. Deep eutectic solvents (DESs) emerge as environmentally friendly alternatives to toxic and volatile organic solvents. One appealing aspect for DESs is that they can be synthesized using naturally occurring compounds from biomass. Herein, we prepared novel hydrophobic DESs based on lignin derivatives and characterized their physicochemical properties including density, viscosity, and thermal behavior. The results showed that five lignin-derived hydrophobic DESs made from menthol, thymol, and 2,6-dimethoxyphenol were promising as green solvents due to their low viscosities and environmentally friendly constituents. To evaluate the potential application …

Recombinant Protein Vaccines Formulated With Enantio-Specific Cationic Lipid R-Dotap Induce Protective Cellular And Antibody-Mediated Immune Responses In Mice, Siva Gandhapudi, Hua Shi, Martin Ward, John Peyton Bush, Margarita G. Avdiushko, Karuna Sundarapandiyan, Lauren V. Wood, Mania Dorrani, Afsheen Fatima, Joe Dervan, Frank Bedu-Addo, Greg Conn, Ted M. Ross, Jerold Woodward

Markey Cancer Center Faculty Publications

Adjuvants are essential components of subunit vaccines added to enhance immune responses to antigens through immunomodulation. Very few adjuvants have been approved for human use by regulatory agencies due to safety concerns. Current subunit vaccine adjuvants approved for human use are very effective in promoting humoral immune responses but are less effective at promoting T-cell immunity. In this study, we evaluated a novel pure enantio-specific cationic lipid 1,2-dioleoyl-3-trimethylammonium-propane (R-DOTAP) as an immunomodulator for subunit vaccines capable of inducing both humoral- and cellular-mediated immunity. Using recombinant protein antigens derived from SARS-CoV2 spike or novel computationally optimized broadly reactive influenza antigen (COBRA) …

Production And Characterization Of Monoclonal Antibodies To Xenopus Proteins, Brett Horr, Ryan Kurtz, Ankit Pandey, Benjamin G. Hoffstrom, Elizabeth Schock, Carole Labonne, Dominique Alfandari

Markey Cancer Center Faculty Publications

Monoclonal antibodies are powerful and versatile tools that enable the study of proteins in diverse contexts. They are often utilized to assist with identification of subcellular localization and characterization of the function of target proteins of interest. However, because there can be considerable sequence diversity between orthologous proteins in Xenopus and mammals, antibodies produced against mouse or human proteins often do not recognize Xenopus counterparts. To address this issue, we refined existing mouse monoclonal antibody production protocols to generate antibodies against Xenopus proteins of interest. Here, we describe several approaches for the generation of useful mouse anti-Xenopus antibodies to multiple …

Grassland Productivity And Water Quality: A 21st Century Issue, David M. Nash, P. M. Haygarth

IGC Proceedings (1977-2023)

1. Irrigation and other changes to the hydrological cycle can increase soil and water salinity.
2. Primary salinisation is a natural process that affects much of Europe, Asia, Africa, the Americas and Australia. Secondary salinisation is caused by human activities such as irrigation and land clearing that mobilise salt stored in the soil.
3. The critical water contaminants exported from grasslands are nitrogen, phosphorus, potential pathogens and sediment.
4. The mechanisms responsible for diffuse pollution from grasslands and mitigation strategies are most effectively investigated using a ‘source-mobilisation-transport’ framework.
5. There is a lack of coherent interaction across discipline boundaries that links pollutant sources to impact. …

Water Resources, Agriculture And Pasture: Implications Of Growing Demand And Increasing Scarcity, M. W. Rosegrant, R. A. Valmonte-Santos, S. A. Cline, C. Ringler, W. Li

IGC Proceedings (1977-2023)

1. Water availability for irrigation is threatened in many regions by rapidly increasing demand for nonagricultural water uses in industry, households, and the environment. The scarcity of irrigation water will not only impact crop production, but also meat production, as much of the pasture used to feed livestock is irrigated.
2. Grassland is caught between two countervailing forces: a requirement for increasing meat demand that boosts the need for additional pasture to support livestock production, and rapidly increasing water scarcity that makes pasture irrigation uneconomical.
3. The most effective means of dealing with water scarcity is likely to be conserving water in existing …

Soil Quality Assessment And Management, M. G. Kibblewhite

IGC Proceedings (1977-2023)

1. Soil quality is related to the capacity of soil to deliver ecosystem services on a sustainable basis.
2. Effective management of soil within grasslands can deliver many benefits to mankind but poor management may cause loss of soil quality from erosion, loss of organic matter, physical deterioration etc.
3. Services are delivered from soil by biological processes. Soil quality depends on the form and condition of the soil habitat. Fixed factors (e.g. texture) are useful for assigning soil to types. Variable factors (e.g. organic carbon) can then be used to assess quality within soil types, by reference to percentiles of the distribution …

Soil Microbial Community: Understanding The Belowground Network For Sustainable Grassland Management, Y. G. Zhu, W. D. Kong, B. D. Chen, Z. B. Nan, P. Christie

IGC Proceedings (1977-2023)

1. In addition to the use of conventional methodologies in soil microbial research, molecular techniques are now being applied to gain insights into the soil microbial community;
2. Plant diversity can exert impacts on soil microbial diversity (through root activities and plant litter etc.), but may in itself be significantly altered by soil properties;
3. Soil microbial diversity largely determines the stability of soil ecosystems under biotic and abiotic perturbations.
4. Management of soil microbial diversity can only be achieved through better understanding their structures and functions.

Grasslands1 For Production And The Environment, David R. Kemp, David L. Michalk

IGC Proceedings (1977-2023)

1. To manage grasslands for production and enhanced environmental values requires a redefinition of the frameworks within which management decisions are made, and a tailoring of practices to suit the ways that farmers operate.
2. Improving the perenniality and permanence of grasslands usually leads to better environmental and production outcomes.
3. There is a case for a more conservative approach to utilising grasslands in order to sustain the functioning of local ecosystems and to improve water quality, nutrient and energy cycling and biodiversity.
4. A landscape rather than paddock focus is more appropriate for meeting current grassland management objectives. Grasslands can be triaged to …

The Potential Of Grassland And Associated Forages To Produce Fibre, Biomass, Energy Or Other Feedstocks For Non-Food And Other Sectors: New Uses For A Global Resource, M. F. Askew

IGC Proceedings (1977-2023)

1. In developed countries increased areas of land will become available for non-food production. Recent reforms of the Common Agricultural Policy will further intensify this trend in Europe.
2. There is potential for grassland and associated species to contribute to large tonnage markets of energy and bulk fibres, to the supply of fermentation products and to speciality markets, but processes and approaches to the market are not as yet developed.
3. There is potential for the establishment of Graminaceous species - specifically for non- food use. For European conditions particular attention is being given to Miscanthus sinensis (Miscanthus), Arundo donax (Giant Reed), Phalaris …

Grasses As Biofactories: Scoping Out The Opportunities, Nick Roberts, K. Richardson, G. Bryan, Christine R. Voisey, W. Mcnabb, T. Conner, M. Christey, R. Johnson

IGC Proceedings (1977-2023)

1. Plant biopharming is set to dominate commercial recombinant protein expression for specific proteins.
2. The choice of plant species depends on a multitude of factors and is determined on a case- by-case basis.
3. As a leaf based expression system grasses would have to compete predominantly with tobacco and alfalfa.
4. The grass-endophyte symbiosis offers a number of unique possibilities for biopharming.

Adoption Of Tropical Legume Technology Around The World: Analysis Of Success, H. M. Shelton, Steven Franzel, M. Peters

IGC Proceedings (1977-2023)

1. Examples of successful adoption of forage legumes are reported from all continents, where they delivered profitability and often provided multipurpose benefits to farmers.
2. Factors vital to successful adoption were: meeting the needs of farmers; building relevant partnerships; understanding the socio-economic context and skills of farmers; participatory involvement with rural communities; and long-term involvement of champions.
3. Organisation of seed supply, achieving scale-up and forming partnerships to implement adoption are key features.
4. Legumes remain an important but under-exploited resource for tropical farming systems. The alternative to legumes will be greater and more costly use of N-fertilisers and purchased protein concentrates.
5. The R&D …

Evolution Of Integrated Crop-Livestock Production Systems, Martin H. Entz, William D. Bellotti, J. M. Powell, S. V. Angadi, W. Chen, K. H. Ominski, B. Boelt

IGC Proceedings (1977-2023)

1. Many factors contribute to changes in the crop-livestock systems, but no logical end-point in the evolution process exists.
2. While benefits of integrated crop-livestock systems over specialised crop and livestock systems are well documented, there has been a move to specialised crop and livestock production.
3. Sustainability issues (manure nutrient concentration, soil quality maintenance, salinity, herbicide resistance, economic instability) have created a renewed interest in integrated crop-livestock systems.
4. Farmer adaptability is as an important link in the evolution between ‘states of integration’.