Livestock Consumption Research Articles

A thematic review on livestock manure treatment strategies focusing on thermochemical conversion.

Livestock manure (LSM) management is emerging as a challenge due to increasing livestock consumption. Owing to the decreased agricultural land area, it is necessary to ensure LSM utilization in non-agricultural fields. LSM can be a valuable resource if managed as a circulating resource. This study discusses research trends based on a literature review and classifies LSM treatments. The analysis of each treatment is presented according to research trends, and implications for the future LSM processing are discussed. "Biological treatment" accounted for the largest portion at 48%, "manure management," which suggests improvement in manure treatment through systematic thinking or LSM management practices, accounted for 16%, and "thermochemical conversion" accounted for 11%. In addition, "life cycle assessment (LCA) research," "solid-liquid separation approach," and "nutrient-recovery/losses" were derived. Studies on biological treatments are increasing. Although anaerobic digestion (AD) is the most used method, it has the disadvantages of long processing time and waste generation after processing. As a key supplement, thermochemical conversion (TCC) technology, which could overcome the disadvantages of AD, was reviewed.

Study of Tiu Kulit Dam Reservoir Operational Pattern Simulation in Sumbawa Island

Tiu Kulit Reservoir plays a pivotal role in managing water resources in West Nusa Tenggara, Indonesia, serving as a vital source for raw water, irrigation, and livestock consumption in the neighboring regions. This study aims to assess the operational dynamics of Tiu Kulit Reservoir across varying hydrological conditions, namely dry, normal, and wet years. The analysis is based on inflow rates and equilibrium water surface elevations. To achieve the objectives, an investigation of the reservoir's operation pattern was conducted. Inflow rates and equilibrium water surface elevations were scrutinized to discern operational behaviors under different hydrological scenarios. The findings indicate distinct operational strategies for Tiu Kulit Reservoir. During dry years, the outflow rate is restricted to 1.20 m3/s at a water surface elevation of +53.07 m. Conversely, the reservoir's operation in normal and wet years allows for a maximum outflow rate aimed at satisfying water demand, set at +57.00 m. In instances of excessive rainfall, the study highlights the potential for overflow beyond the spillway threshold. The study underscores the significance of these results for effective water management and resource allocation in the region. Understanding the reservoir's behavior under various conditions enables informed decision-making and sustainable utilization of water resources. To build upon this study, it is recommended to implement continuous monitoring systems for both inflow and outflow rates. Furthermore, the installation of rainfall monitoring stations within the catchment area would enhance data accuracy. Regular evaluations of the reservoir's operational patterns are also advised, allowing for adaptive strategies in response to evolving climate conditions. Moreover, future research endeavors should focus on augmenting water supply to meet the demands of a growing population and livestock, while optimizing agricultural practices to boost productivity.

Global patterns and correlates in the emergence of antimicrobial resistance in humans.

Antimicrobial resistance (AMR) is a critical global health threat, and drivers of the emergence of novel strains of antibiotic-resistant bacteria in humans are poorly understood at the global scale. We examined correlates of AMR emergence in humans using global data on the origins of novel strains of AMR bacteria from 2006 to 2017, human and livestock antibiotic use, country economic activity and reporting bias indicators. We found that AMR emergence is positively correlated with antibiotic consumption in humans. However, the relationship between AMR emergence and antibiotic consumption in livestock is modified by gross domestic product (GDP), with only higher GDP countries showing a slight positive association, a finding that differs from previous studies on the drivers of AMR prevalence. We also found that human travel may play a role in AMR emergence, likely driving the spread of novel AMR strains into countries where they are subsequently detected for the first time. Finally, we used our model to generate a country-level map of the global distribution of predicted AMR emergence risk, and compared these findings against reported AMR emergence to identify gaps in surveillance that can be used to direct prevention and intervention policies.

Effect of Consumption of Animal Products on the Gut Microbiome Composition and Gut Health.

The gut microbiome is critical in human health, and various dietary factors influence its composition and function. Among these factors, animal products, such as meat, dairy, and eggs, represent crucial sources of essential nutrients for the gut microbiome. However, the correlation and characteristics of livestock consumption with the gut microbiome remain poorly understood. This review aimed to delineate the distinct effects of meat, dairy, and egg products on gut microbiome composition and function. Based on the previous reports, the impact of red meat, white meat, and processed meat consumption on the gut microbiome differs from that of milk, yogurt, cheese, or egg products. In particular, we have focused on animal-originated proteins, a significant nutrient in each livestock product, and revealed that the major proteins in each food elicit diverse effects on the gut microbiome. Collectively, this review highlights the need for further insights into the interactions and mechanisms underlying the impact of animal products on the gut microbiome. A deeper understanding of these interactions would be beneficial in elucidating the development of dietary interventions to prevent and treat diseases linked to the gut microbiome.

A One Health Initiative For Air Pollution: Student-Living Gardens

Air pollution is one of the largest issues facing our planet to date. It leads to a variety of severe consequences including an increased incidence of respiratory illness in humans and non-human animals, damage to plants, and exacerbation of climate change. An enormous contributor to air pollution is the livestock farming industry which, in addition to its negative environmental impacts, detrimentally affects the mental health and well-being of non-human animals through various unnatural practices. However, air pollution may be mitigated by planting gardens at homes located in the student-living area of Queen’s University in Kingston, Ontario. These gardens would include vegetables, low-maintenance plants, and edible native species which would remove toxins from the air and provide multiple additional benefits to humans, non-human animals, and the environment. One of the greatest benefits of the proposed gardens would be the provision of vegetables and edible native species, allowing students to consume more plant-based foods and stray away from livestock consumption. The gardens would also increase ecosystem biodiversity, which would not only make plant life more resilient but also help create new opportunities for reliable food sources and habitats. If the success of the proposed initiative were to be proven within the area, additional strategies may be introduced in Kingston to further reduce air pollution and perhaps inspire other university communities to undergo similar changes.

For a Better Quality of Beef: The Challenge from Growing Livestock on Limited Grasslands with a Production-Consumption Balance Perspective.

The growing population, the transition dietary towards animal-based products, and the preference for the brand of grass-feeding livestock are bringing increasing pressure on natural grasslands, especially for dry-land areas. The Xilingol League of China is famous for its free-range livestock product, however, overgrazing and herders' benefits damage are always serious issues for this semi-arid grassland region. This study focuses on the relationship between the supply of natural grassland and the consumption of free-range livestock in the Xilingol League, and this study employed the grassland carrying capacity as the index to judge the sustainability states and its trends of the local grass-feeding system. Satellite data production of net primary production was used for grassland production, statistical livestock data and the consumption model were used for actual forage consumption, and empirical key informant interview data were used to obtain a more comprehensive understanding. The results show that: (1) the natural grassland carrying capacity of the Xilingol League fluctuated, showing improvement from 2000 to 2021; (2) the grassland management needs to be more diversified in different regions with different natural conditions; and (3) while the demand for free-range, high-quality beef is increasing, attention should be paid to the carrying capacity of natural pastures and more consideration should be taken of the benefits of balancing the livelihood of herders, policy strategies, and the customers' preferences. Potential ways of doing this include employing technologies to improve livestock production, and further exploring and promoting the economic value of the free-range livestock and the geographical indication to get the economic-ecological win-win situation. The research framework and results would be beneficial to reveal the potential threats in pastoral areas and provide support for the optimization of the regional grass-feeding breeding system, especially in middle-income countries.

The soil-borne fungal pathogen Athelia rolfsii: past, present, and future concern in legumes.

Legumes are ubiquitous, low-cost meals that are abundant in protein, vitamins, minerals, and calories. Several biotic constraints are to blame for the global output of legumes not meeting expectations. Fungi, in particular, are substantial restrictions that not only hinder production but also pose a serious health risk to both human and livestock consumption. Athelia rolfsii (Syn. Sclerotium rolfsii) is a dangerous pathogenic fungus that attacks most crops, causing massive yield losses. Legumes are no longer immune to this dreadful fungus, which can potentially result in a 100% yield loss. The initial disease symptoms based on the formation of brown color lesions at the point of infection and further development of mycelia, followed by yellowing and wilting of the whole plant. To tackle such situation, various strategies, i.e., management in cultural practices, disease-free plant growth, genetic changes, crop hybridization and in vitro culture techniques have been undertaken. This present review encapsulates the entire situation, from sclerotial dissemination through infection development and control in legume crops, with the goal of developing a tangible understanding of sustainable legume production improvements. Further study in this area might be led in an integrated manner as a result of this information, which could contribute to a better understanding of the processes of disease incidence, resistance mechanism, and its control, and fostering greater inventiveness in the production of legumes.

Groundwater Management for Agricultural Purposes Using Fuzzy Logic Technique in an Arid Region

The study aimed to determine groundwater’s suitability for irrigation and cattle rearing in Kuwait. In this regard, groundwater samples were collected from Umm Al Aish (UA) and adjoining Rawdhatain (RA) water wellfields to develop groundwater suitability maps for irrigation purposes using the fuzzy logic technique in ArcGIS. RA was dominated by Na-Cl, Na-Ca, and Ca-SO4 water types, whereas UA was dominated by the Ca-Mg water type. Due to the influence of the temperature and pCO2, the carbonates were inferred to be more susceptible to precipitation in the soil than the sulfates. The ternary plots for both regions revealed that the samples’ suitability ranged from good to unsuitable. Spatial maps of nine significant parameters governing the irrigation suitability of water were mapped and integrated using the fuzzy membership values for both regions. The final suitability map derived by overlaying all the considered parameters indicated that 8% of the RA region was categorized as excellent, while UA showed only 5%. Samples situated in the study areas showed an excellent to very satisfactory range for livestock consumption. Developing a monitoring system along with innovative water resource management systems is essential in maintaining the fertility of the soil and existing groundwater reserves.

Understanding the mechanism of pathogenicity through interactome studies between Arachis hypogaea L. and Aspergillus flavus

Aspergillus flavus (A. flavus) infects the peanut seeds during pre-and post-harvest stages, causing seed quality destruction for humans and livestock consumption. Even though many resistant varieties were developed, the molecular mechanism of defense interactions of peanut against A. flavus still needs further investigation. Hence, an interologous host-pathogen protein interaction (HPPI) network was constructed to understand the subcellular level interaction mechanism between peanut and A. flavus. Out of the top 10 hub proteins of both organisms, protein phosphatase 2C and cyclic nucleotide-binding/kinase domain-containing protein and different ribosomal proteins were identified as candidate proteins involved in defense. Functional annotation and subcellular localization based characterization of HPPI identified protein SGT1 homolog, calmodulin and Rac-like GTP-binding proteins to be involved in defense response against fungus. The relevance of HPPI in infectious conditions was assessed using two transcriptome data which identified the interplay of host kinase class R proteins, bHLH TFs and cell wall related proteins to impart resistance against pathogen infection. Further, the pathogenicity analysis identified glycogen phosphorylase and molecular chaperone and allergen Mod-E/Hsp90/Hsp1 as potential pathogen targets to enhance the host defense mechanism. Hence, the computationally predicted host-pathogen PPI network could provide valuable support for molecular biology experiments to understand the host-pathogen interaction. SignificanceProtein-protein interactions execute significant cellular interactions in an organism and are influenced majorly by stress conditions. Here we reported the host-pathogen protein-protein interaction between peanut and A. flavus, and a detailed network analysis based on function, subcellular localization, gene co-expression, and pathogenicity was performed. The network analysis identified key proteins such as host kinase class R proteins, calmodulin, SGT1 homolog, Rac-like GTP-binding proteins bHLH TFs and cell wall related to impart resistance against pathogen infection. We observed the interplay of defense related proteins and cell wall related proteins predominantly, which could be subjected to further studies. The network analysis described in this study could be applied to understand other host-pathogen systems generally.

Genetic Diversity of Toxoplasma gondii by Serological and Molecular Analyzes in Different Sheep and Goat Tissues in Northeastern Iran.

Toxoplasmosis is a parasitic disease caused by compilation protozoan agent Toxoplasma gondii, leading to significant financial and quality-adjusted life-year losses. Overcooked or raw meat consumption has been a considerable transmission route. The present study was conducted to determine the seropositivity rate of T. gondii in sheep and goats by serological and molecular tests and genotyping of obtained isolates in northeast Iran. Blood and tissue samples (diaphragm, heart) of 296 animals (including 168 sheep and 128 goats) were collected from the slaughterhouse in Quchan Country from august 2016 to April 2017. Modified agglutination test (MAT) and the PCR method performed to detect parasite DNA on tissues.PCR-RFLP method of GRA6 gene was used to determine the genotype of T. gondii. In addition, sequencing analysis was performed to evaluate the Toxoplasma type strains. Serum positive for MAT results were found in 27.4% of sheep and 23.4% of goats. Positive PCR of B1 gene results in diaphragm and heart tissues of sheep and goats was 47.8% and 26.1%, 40% and 23.3%, respectively. PCR of GRA6 gene results were positive in 10 samples that RFLP technique results using MseI enzyme revealed genotype I. Sequencing and phylogenetic analysis revealed DNA of all samples was closely related to Toxoplasma type I. Concerning the high seropositivity rate of toxoplasmosis, undertaking an appropriate preventive program for reducing the prevalence of T. gondii infection by raw or undercooked meat consumption of livestock is recommended. Our study supports the notion that these animals' consumption of raw and undercooked meat can be a probable source of human toxoplasmosis.

Effectiveness and implementation challenges of a livestock asset transfer intervention for smallholders in community-protected areas in Cambodia.

Limited opportunities to generate income in community-protected areas (CPAs) often lead to the illegal harvesting of wildlife and natural resources, undermining the aim of protected areas. Sustained livestock production can be an alternative income source. To understand the effectiveness and feasibility of livestock production in CPAs. We conducted a livestock asset transfer intervention in 25 CPAs across three agroecological zones in Cambodia. We monitored livestock mortality, consumption and sales of livestock over 2 years. Participant observations and structured questionnaires were conducted to elicit information about constraints for livestock production perceived by the participants. A total of 756 households were recruited and 320, 184 and 252 households received chicken, pigs and cattle, respectively. All participants received technical training in livestock production and biosecurity management practices. After the intervention, the number of chickens, pigs and cattle increased on average by 5.9 (range: 0.3-26.3), 0.5 (-1 to 2.7) and 0.12 (0-0.35) per 1 input animal, respectively. The extent of increase was significantly different between zones only for chickens (Kruskal-Wallis test p=0.004). The number of chickens and pigs sold per household was significantly different among zones. We observed that training was ineffective to alter livestock management practices in some CPAs, which partially explains their suboptimal performance of livestock production. Understanding contextual factors required for successful livestock production in CPAs is crucial for improving livelihoods and preventing biodiversity loss in Cambodia.

Alternaria gansuense, a Plant Systematic Fungal Pathogen Producing Swainsonine in Vivo and in Vitro.

Astragalus adsurgens (A. adsurgens), which is considered a forage in China, grows widely in Eurasia and North America. However, Alternaria gansuense (A. gansuense) (synonym: Embellisia astragali) systematically infects A. adsurgens, producing swainsonine (SW), which poisons domesticated animals. In this study, we hypothesized that the A. gansuense SW-producing fungus is morphologically and molecularly related to the locoweed endophyte, Alternaria oxytropis (A. oxytropis), which systematically grows in host plants. Therefore, pure cultures of the fungi from diseased plants or endophytic interactions were collected from fields and assayed for SW via high-performance liquid chromatography linked to mass spectroscopy (HPLC-MS). The production of SW was also detected in A. adsurgens, A. oxytropis and diseased plants by assaying for the presence of the β-ketoacyl synthase (KS) gene, which is required for SW synthesis. Diseased A. adsurgens and pure cultures of A. gansuense have SW and the healthy-looking A. adsurgens plants also contained SW, probably because they were infected with A. gansuense. Therefore, A. adsurgens-infected A. gansuense are not safe for livestock consumption.

Pengaruh Teknik Silase Pakan Komplit Tersuplementasi Ampas Tahu Berprotein Kasar Berlebih pada Domba Secara InVivo

Tofu dregs are feed ingredients with high nutritional content and easy to rot. Therefore a silage storage technique is needed to preserve and reduce environmental pollution. The process of preserving high-protein feed by silage has a weakness in the process of lowering the pH. This happens because the ammonia from the deamination process is alkaline . The silage processing can converting some nutrients into organic acids which can improve livestock health and production. The Complete feed silage which was supplemented with tofu dregs was carried out on thin-tailed local rams aged 8-10 months and body weight 16-18 kg. The rams were grouped to 4 groups. The first gorup was P0 = complete feed which was suplemented with tofu dregs without silage processing compared to P1 = P0 + silage processing which was fermented for at least 14 days. The data then analyzed using t-independent method in a randomized block design on the basis of body weight. The results of the study showed that the average dry matter consumption of livestock was not significantly different P>0.1, the increase in monthly weight was higher in group P1 with a P value <0.05, the silage process increased the efficiency of p<0.05. Both treatment had no effect on cholesterol, blood urea nitrogen (BUN), glucose, red blood grains and white blood grains. So it can be concluded that feeding by silage is better than fresh
 
 .
 Keywords: Silage, rams, tofu dregs
 

Characterization of the water bodies of Extremadura (SW Spain)

Extremadura is the region that stores the greatest amount of fresh water in Spain. Such water is mainly used for power generation, irrigation in agriculture, biodiversity conservation, tourism, recreation, and human and livestock consumption. Nevertheless, crucial information on the total number of water bodies and their geometrical characteristics and spatial distribution patterns are still missing. Thus, our main goal was to characterize the Extremenian water bodies geometrically and spatially through different statistical techniques such as kernel density, Moran’s index, the Getis-Ord Gi*, and principal component analysis (PCA). Firstly, all existing hydrological information was gathered, and using aerial aircraft imagery and satellite images, each water body (WB) was then carefully collected, checked, and corrected. We have inventoried 100,614 WBs (mean density: 2.45 WB km2), irregularly distributed on the territory. WBs with an area < 0.01 km2 (100 ha) represent 64.5% of the total. A multivariate statistical study was conducted, showing that livestock, aridity of the climate, and topography are the main factors controlling the density of water bodies (WBs) in this area. It can be concluded that monitoring of small bodies is crucial to understand their spatial distribution, since they are spread over areas in which extensive farming and commercial crops such as tobacco strongly influence the way of living of many families.

Potential of Market Vegetable Waste as Animal Feed in Greater Malang

This study aims to determine the potential of vegetable waste as animal feed from various traditional markets in the city of Malang. This research is descriptive and experimental. The descriptive method is used to overview the diversity, the amount of availability of various vegetable waste for each season, the type of vegetable waste that dominates the market that is suitable for livestock consumption, and the nutritional value of vegetable waste. The experimental research method used a randomized block design ANOVA with an accuracy of 1% to determine the chemical quality of vegetable waste that dominates several markets and the chemical quality of vegetable waste based on shelf life with observations of 0, 2, 4, and 6 days of storage. The results showed that the accumulation of vegetable waste per day in the selected market was 3774.76 kg in dry matter, equivalent to meeting the consumption needs of 308.14 ruminant units. The conclusion is that the nutritional value of vegetable waste from the market is the same as the nutritional value of elephant grass from DM, OM, CP, DMD, and OMD. Vegetable waste from the market easily loses its nutritional value as a result of the decay process.

Formulation of Criminal Law Policy for Distribution of Non-Food Animal Meat for Consumption

This study aims to provide an analysis of the many cases of distribution of non-food animal meat for consumption purposes that have recently received attention, such as the rampant sale of cat and dog meat which often occurs in society. Cats and dogs are pets kept by humans. These animals are not included in the consumption of livestock. Consumption of non-food animals is a heinous act that can become a zoonotic vector. This study uses normative legal research methods with several approaches, namely the statutory approach and the conceptual approach. The results obtained are that there are laws and regulations related to the distribution of non-food animal meat for consumption purposes, but some of these regulations do not mention the imposition of sanctions on the distribution of non-food animal meat for consumption purposes. Criminal laws should provide for the torture of animals and the distribution of non-food animal meat for consumption and sanctions. This is considering the dangers of consuming non-food animals which have the potential to harm the wider community.How to cite item: Sabrina, Nahdiya. “Formulation of Criminal Law Policy for Distribution of Non-Food Animal Meat for Consumption.” Jurnal Cakrawala Hukum. 14 no. 1 (2023): 28-35. DOI: 10.26905/idjch.v14i1.10049.

Global Food Production and Distribution Analysis using Data Mining and Unsupervised Learning.

Today's food industry is extensive and complicated, encompassing anything from subsistence agriculture to multinational food corporations. The mobility of food and food elements in food systems has a major impact on biodiversity preservation and the overall sustainability of our fragile global ecosystem. Identifying the human and livestock consumption patterns across regions and territories will optimize the dietary standards of the habitually undernourished and the expanding population without substantially increasing the amount of land under cultivation. Food preservation is the basis for economic advancement and social sustainability, so the food industry, both local and global, is fundamental to everyone. As a primary mechanism for ensuring global food preservation, there is currently a strong emphasis on accelerating food supply and decreasing waste. Thus, analyzing the production and distribution of food supply will boost economic sustainability. In this paper, we present a quantitative analysis of global and regional food supply to reveal the flow of food and feed products in various parts of the world. Using data mining and machine learning-based approaches, we seek to quantify the production and distribution of food elements. The study aims to employ artificial intelligence-based methods to comprehend the shift and change in supply and consumption patterns with timely distribution to meet the global food instability. The method involves using statistical-based approaches to identify the hidden factors and variables. Feature engineering is used to uncover the interesting features in the dataset, and various clustering-based algorithms, like K-Means, have been utilized to group and identify the similar and most notable features. The concept of data mining and machine learning-based algorithms has helped us in identifying the global food production and distribution subsystem. The identified elements and their relationship can help stakeholders in regulating various external and internal factors, including urbanization, urban food needs, the economic, political and social framework, food demand, and supply flows. The exploratory analysis helps in establishing the efficiency and dynamism of food supply and distribution systems. The outcome demonstrates a pattern indicating the flow of currently grown crops into various endpoints. Few countries with massive populations have shown tremendous growth in their production capacity. Despite the fact that only a few countries produce a large portion of food and feed crops, still it is insufficient to feed the estimated global population. Significant changes in many people's socioeconomic conditions, as well as radical dietary changes, will also be required to boost agricultural credit and economic foundations.

Development and validation of the nutrient-rich food index for China

To select the nutrient-rich food(NRF) index applicable to the food characteristics of Chinese residents through population-based dietary quality validation. Sixteen NRF indices were constructed based on the Chinese Food Composition Table and the Chinese Food Labelling Nutrient Reference Values, using 100 g as the base amount of food for calculation. The NRF indices were validated by the China Prime Diet Quality Score(CPDQS), a dietary quality assessment index, using food intake data collected from 14 068 adult residents in 15 provinces in 2018 under the China Health and Nutrition Survey. The CPDQS was used as the dependent variable and the dietary NRF index score was used as the independent variable. The multiple linear regression models were performed after adjusting the covariates and the NRF index with the largest coefficient of determination R~2 was used as the most appropriate index. Regression analysis showed that the R~2 of NRF8.2 index was the largest, which was 0.16. Dried legumes, fungi and algae, nuts, vegetables, eggs and aquatic products scored higher on the NRF8.2 index. The high scoring subgroups had higher average daily intakes of cereals and potatoes, vegetables, fruits, soybeans and nuts, dairy, eggs and aquatic products, as well as lower consumption of livestock and poultry meat, beverages, salt and cooking oil, compared to the low scoring subgroup on the Dietary NRF8.2 Index. In terms of nutrient intake, the medium and high NRF8.2 subgroups had relatively high intakes of energy, protein, vitamins and minerals and relatively low intakes of fat and sodium. The NRF8.2 index explains the maximum variation in CPDQS and is a relatively suitable NRF index for the Chinese population.

감압 텀블링 해동이 돈육의 품질 특성에 미치는 영향

As livestock consumption in Korea has been gradually increasing, the quality of the final products has been improved to meet this increased demand. In particular, maintaining the water holding capacity (WHC) and minimizing the drip loss during the thawing of frozen meat are of utmost importance. This study investigated the physicochemical properties of frozen pork subjected to thawing under different conditions: at room temperature (20°C, under air), at a low temperature (4°C refrigerator, under air), under water (20°C, under water in a vacuum bag), under microwave (microwave-thawing, 260 W), and under low-pressure tumbling (20°C, 0.015 bar, tumbling). The shortest thawing time for frozen pork was recorded upon low-pressure tumbling thus indicating a fast heat transfer. The lowest drip loss (0.2%) and highest WHC (94.5%) were also recorded under this condition. A significantly higher drip loss was observed upon microwave-(1.0%) and water-thawing (1.2%), which resulted in the lowest WHC in microwave thawing (87.2%). The highest total count of aerobic bacteria and coliform group were observed upon room temp thawing while the low pressure tumbling and thawing resulted in the lowest aerobic bacteria (1.90 log CFU/g) and coliform (0.78 log CFU/g) count. Consequently, thawing by low pressure tumbling afforded the best food quality.

Multiplex CRISPR/Cas9-mediated raffinose synthase gene editing reduces raffinose family oligosaccharides in soybean.

Soybean [Glycine max (L.) Merr.] is an important world economic crop. It is rich in oil, protein, and starch, and soluble carbohydrates in soybean seeds are also important for human and livestock consumption. The predominant soluble carbohydrate in soybean seed is composed of sucrose and raffinose family oligosaccharides (RFOs). Among these carbohydrates, only sucrose can be digested by humans and monogastric animals and is beneficial for metabolizable energy, while RFOs are anti-nutritional factors in diets, usually leading to flatulence and indigestion, ultimately reducing energy efficiency. Hence, breeding efforts to remove RFOs from soybean seeds can increase metabolizable energy and improve nutritional quality. The objective of this research is to use the multiplex Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9-mediated gene editing system to induce the knockout of soybean raffinose synthase (RS) genes RS2 and RS3 simultaneously to reduce RFOs in mature seeds. First, we constructed five types of multiplex gene editing systems and compared their editing efficiency in soybean hairy roots. We confirmed that the two-component transcriptional unit (TCTU) and single transcriptional unit (STU) systems with transfer RNA (tRNA) as the cleavage site performed better than other systems. The average editing efficiency at the four targets with TCTU-tRNA and STU-tRNA was 50.5% and 46.7%, respectively. Then, we designed four single-guide RNA (sgRNA) targets to induce mutations at RS2 and RS3 by using the TCTU-tRNA system. After the soybean transformation, we obtained several RS2 and RS3 mutation plants, and a subset of alleles was successfully transferred to the progeny. We identified null single and double mutants at the T2 generation and analyzed the seed carbohydrate content of their progeny. The RS2 and RS3 double mutants and the RS2 single mutant exhibited dramatically reduced levels of raffinose and stachyose in mature seeds. Further analysis of the growth and development of these mutants showed that there were no penalties on these phenotypes. Our results indicate that knocking out RS genes by multiplex CRISPR/Cas9-mediated gene editing is an efficient way to reduce RFOs in soybean. This research demonstrates the potential of using elite soybean cultivars to improve the soybean meal trait by multiplex CRISPR(Clustered Regularly Interspaced Short Palindromic Repeats)/Cas9-mediated gene editing.