Effects of Used Battery on Key Enzyme Activity during the Germination of Wheat Seeds

[Objective] This study aimed to explore the effects of used battery lixivium on wheat germination. [Method] The wheat seeds were treated with used battery lix- ivium at different concentrations to detect the change of activities of amylase, pro- tease, pyruvate dehydrogenase （PDH） and polyphenol oxidase （PPO） during the ger- mination period. [Result] The results showed that the used battery affected enzyme activity. With the increase of concentration of used battery lixivium, trends of the changes of amylase and protease activities were not different. The activities were en- hanced at low concentrations of lixivium, while were inhibited at high concentrations. The tends of changes of pyruvate dehydrogenase （PDH） and polyphenol oxidase （PPO） activities were not consistent with that of either amylase or protease, which showed continuous downward trends with the increasing concentration of used battery lixivium. [Conclusion] This study is of great practical significance for understanding the effects of used battery lixivium on the germination of wheat seeds.

Responses of Nitrogen and Related Enzyme Activities to Fertilization in Rhizosphere of Wheat

In the present experiment, wheat seedlings (Trticucum aestivum L.) were grown on a purple soil withvarious fertilizer treatments in order to investigate the responses of nitrogen and related eusyme activitiesin the rhizosphere. The results revealed the increments of both organic matter and total N in the soil withthe proximity to the growing roots, especially in the treatment of supplying pig manure in combination withchemical fertilizer, suggesting that they could come from root and microorganism exudation which could beintensified by inorganic-organic fertilization, being of benefit to improving the physical and biological envi-ronment in the rhizosphere of wheat. Much more inorganic N was observed in the fertilized soils surroundingwheat roots than in the CK treatment, indicating the improvement of crop N supply in the rhisosphere ofwheat by fertilization.The activities of invertase, urease and protease in the root zone were greatly enhanced as compared tothose in the other parts of soils except that the urease activities were similar in the rhizo8phere and nonrhi-zosphere of the CK and pig manure treatments, indicating that invertase and protease could be produced bygrowing roots and rhizosphere microorganisms, in contrast to urease which could be stimulated by urea. Also,significant increment of these three enzyme activities in the rhizosphere of wheat by fertilizatioll, especially byapplication of chemical fertilizer combined with pig manurel suggested that fertilization not only acceleratedthe renewal of organic matter but also enhanced bioavailability of organic N in that soil zone. This couldbe the reason why the total amount of inorganic N available for plants was increased more obviously in therhizosphere of wheat of the fertilization treatments than in that of the CK treatment.

Wettability changes of wheat straw treated with chemicals and enzymes

A study was conducted to test wettability changes of the wheat straw treated with different methods for the preparation of wheat straw particle board. The wheat straws were separately sprayed with two chemicals （0.6% NaOH, 0.3% H2O2） and three enzymes （lipase, xylanase, cellulase）. The contact angle between water and the surface of wheat straw was measured and the spreading-penetration parameters （K-values） were also calculated with wetting model. The surfaces of treated wheat straw and control sample were scanned by means of Micro-FTIR, and their peaks arrangements were analyzed. The surface morphologies of treated wheat straw and control sample were also observed by SEM. Chemical etching was found on the exterior surfaces of the straws treated separately with 0.6% NaOH and 0.3% H2O2; furthermore, the spreading-penetration parameters （K-values） of the distilled water on the exterior surfaces of the treated wheat straw along the grain were higher than that of control. The wettability of exterior surfaces of the wheat straws treated separately with lipase, xylanase and cellulose were improved after treating for seven days, and among the three enzymes treatments, the lipase treatment showed best result. The lipase treatment and NaOH treatment were determined as better methods for improving the wettability of wheat straw surfaces. However, in the economic aspect, NaOH treatment was more practical and easier in the pretreatment for the manufacture of straw particle board.

Role of renin-angiotensin system/angiotensin converting enzyme-2 mechanism and enhanced COVID-19 susceptibility in type 2 diabetes mellitus

Coronavirus disease 2019(COVID-19)is a disease that caused a global pandemic and is caused by infection of severe acute respiratory syndrome coronavirus 2 virus.It has affected over 768 million people worldwide,resulting in approx-imately 6900000 deaths.High-risk groups,identified by the Centers for Disease Control and Prevention,include individuals with conditions like type 2 diabetes mellitus(T2DM),obesity,chronic lung disease,serious heart conditions,and chronic kidney disease.Research indicates that those with T2DM face a hei-ghtened susceptibility to COVID-19 and increased mortality compared to non-diabetic individuals.Examining the renin-angiotensin system(RAS),a vital regulator of blood pressure and pulmonary stability,reveals the significance of the angiotensin-converting enzyme(ACE)and ACE2 enzymes.ACE converts angiotensin-I to the vasoconstrictor angiotensin-II,while ACE2 counters this by converting angiotensin-II to angiotensin 1-7,a vasodilator.Reduced ACE2 exp-ression,common in diabetes,intensifies RAS activity,contributing to conditions like inflammation and fibrosis.Although ACE inhibitors and angiotensin receptor blockers can be therapeutically beneficial by increasing ACE2 levels,concerns arise regarding the potential elevation of ACE2 receptors on cell membranes,potentially facilitating COVID-19 entry.This review explored the role of the RAS/ACE2 mechanism in amplifying severe acute respiratory syndrome cor-onavirus 2 infection and associated complications in T2DM.Potential treatment strategies,including recombinant human ACE2 therapy,broad-spectrum antiviral drugs,and epigenetic signature detection,are discussed as promising avenues in the battle against this pandemic.

Angiotensin-converting enzyme 2 alleviates liver fibrosis through the renin-angiotensin system

The present letter to the editor is related to the study titled‘Angiotensin-converting enzyme 2 improves liver fibrosis in mice by regulating autophagy of hepatic stellate cells’.Angiotensin-converting enzyme 2 can alleviate liver fibrosis by regulating autophagy of hepatic stellate cells and affecting the renin-angiotensin system.

Effects of High Temperature on Antioxidant Enzymes and Lipid Peroxidation in Flag Leaves of Wheat During Grain Filling Period

On the basis of the phytotron, the effects of high temperature （daily average temperature 25, 30, 35 and 40℃, respectively） on antioxidant enzymes and lipid peroxidation in flag leaves of wheat at 50% relative air moisture during grain fastest filling stage [19-21 days after anthesis （DAA）] were studied. The wheat cultivars tested were Yangmai 9 with weak-gluten and Yangmai 12 with medium-gluten. Compared with 25℃, the higher the temperature was, the higher was the MDA content in flag leaves, while lower were the SOD, POD, and CAT activities. SOD and CAT activities in Yangmai 12 appeared to be more sensitive to high temperature than that in Yangmai 9. But POD activity in Yangmai 12 was less sensitive to high temperature. MDA content in Yangmai 12 was higher than that in Yangmai 9. The 1000-grain weight declined with increase in temperature.

Changes in Activities of Antioxidant-Related Enzymes in Leaves of Resistant and Susceptible Wheat Inoculated with Rhizoctonia cerealis

In this study, the changes in activities of peroxidase （POD）, glutathione S-transferase （GST）, catalase （CAT）, and superoxide distmuase （SOD） in leaves of four wheat cultivars, including resistant cultivars Shannong 0431 and Shanhongmai, a mildsusceptible cultivar Lumai 21, and a susceptible cultivar Wenmai 6, were investigated to explore if these antioxidantrelated enzymes were involved in wheat defense response to Rhizoctonia cerealis attack. Results showed that the activities of these enzymes in different wheat cultivars varied greatly after challenging with R. cerealis. The POD activities in resistant cultivars Shannong 0431 and Shanhongmai increased much quicker after the inoculation and displayed much higher activity than that in susceptible cultivar Wenmai 6. In contrast, POD activity showed a slow accumulation and a delayed peak time. The activities of GST and CAT in Shannong 0431 and Lumai 21 increased obviously within 48 h post inoculation （hpi）, whereas SOD activity decreased dramatically within 24 hpi in Shannong 0431 and 24-48 hpi in Lumai 21. In susceptible cultivar Wenmai 6, the SOD and CAT activities didn’t change obviously, whereas the activity of GST was decreased. The activities of CAT and GST in Shanhongmai did not show significant difference compared with those in Wenmai 6. Results suggested that POD activity may be positively associated with wheat defense to R. cerealis.

Cloning and Phylogenetic Analysis of Ubiquitinconjugating Enzyme Gene TaUBC4 in Different Wheat Cultivars

[Objective] This study aimed to clone ubiquitin-conjugating enzyme gene TaUBC4 from different wheat cultivars and thus analyze their phylogenetic relationship.[Method] The UBC4 coding sequences were cloned through reverse transcription PCR （RT-PCR） from 21 wheat varieties.After sequencing,the UBC4 sequence in wheat cultivar Zhongguochun （GenBank accession No：M28059） was selected as the reference gene,to analyze the mutation frequency and evolutionary distance in the CDSs and corresponding amino acid sequences of the different wheat cultivars.Moreover,the phylogenetic tree based on the amino acid sequences of these TaUBC4 genes were constructed,involving the homologous sequences of TaUBC4 in eight other monocots.[Result] TaUBC4 sequence was highly conserved because the similarity in DNA sequences of the wheat varieties was over 94％,while that in amino acid sequence was over 96％.And the amino acid sequence difference only can be seen at two sites among some varieties.Phylogenetic tree constructed revealed the evolutionary relationships among these wheat varieties.[Conclusion] This study reveals the polymorphism and evolutionary characteristics in the nucleotide and amino acid sequences in different wheat varieties,which lays foundation for investigating the evolution and biological function of TaUBC4 gene.In addition,the phylogenetic tree constructed provides theoretical references for the classification of the wheat varieties with complicated background.

Activities of the Enzymes Involved in Starch Synthesis and Starch Accumulation in the Grains of Wheat Cultivars, GC8901 and SN1391

Two wheat cultivars, GC8901 (hard winter wheat) and SN1391 (soft winter wheat), were used for investigating the changes of enzyme activities for sucrose metabolism and starch biosynthesis and the accumulation character of starch composition. The result showed that activities of sucrose (SS), sucrose-phosphate synthase (SPS), adenosine diphosphorate glucose pyrophrylase (AGPase) and soluble starch syntheses (SSS) of 1391, which have more starch, were significant higher than those of 8901, that with low starch content. But the changing of granule-bound starch synthase (GBSS) activity was consistent with the amylose content, which indicated that amylose contents in grain were determined by GBSS activity, especially the activity at later grain filling stages. Simulating with Richards equation showed that it was initiating time and accumulation rate, but not accumulation duration that determined the content of starch composition. Furthermore, changing of sucrose transport capacity was consistent with SSS and GBSS activities, starch accumulation rate was accordant to AGPase and SS/SPS ration, not SS, SPS, SSS or GBSS activities. The results suggested that there was no inevitable relation of starch accumulating rate and starch composition contents with the activity of single enzyme such as SS, SPS, SSS or GBSS, but closely related to AGPase activity and SS/SPS ratio, and it was SPS and AGPase that play a vital role in the biosynthetic pathway. Later polymerization reactions catalyzed by SSS and GBSS don’t seem to control the rate of starch accumulation, but do affect starch structure.

Hepatic angiotensin-converting enzyme 2 expression in metabolic dysfunction-associated steatotic liver disease and in patients with fatal COVID-19

BACKGROUND Metabolic dysfunction-associated steatotic liver disease(MASLD),characterised by hepatic lipid accumulation,causes inflammation and oxidative stress accompanied by cell damage and fibrosis.Liver injury(LI)is also frequently reported in patients hospitalised with coronavirus disease 2019(COVID-19),while preexisting MASLD increases the risk of LI and the development of COVID-19-associated cholangiopathy.Mechanisms of injury at the cellular level remain unclear,but it may be significant that severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)which causes COVID-19,uses angiotensin-converting expression enzyme 2(ACE2),a key regulator of the‘anti-inflammatory’arm of the renin-angiotensin system,for viral attachment and host cell invasion.AIM To determine if hepatic ACE2 levels are altered during progression of MASLD and in patients who died with severe COVID-19.METHODS ACE2 protein levels and localisation,and histological fibrosis and lipid droplet accumulation as markers of MASLD were determined in formalin-fixed liver tissue sections across the MASLD pathological spectrum(isolated hepatocellular steatosis,metabolic dysfunction-associated steatohepatitis(MASH)+/-fibrosis,end-stage cirrhosis)and in post-mortem tissues from patients who had died with severe COVID-19,using ACE2 immunohistochemistry and haematoxylin and eosin and picrosirius red staining of total collagen and lipid droplet areas,followed by quantification using machine learning-based image pixel classifiers.RESULTS ACE2 staining is primarily intracellular and concentrated in the cytoplasm of centrilobular hepatocytes and apical membranes of bile duct cholangiocytes.Strikingly,ACE2 protein levels are elevated in non-fibrotic MASH compared to healthy controls but not in the progression to MASH with fibrosis and in cirrhosis.ACE2 protein levels and histological fibrosis are not associated,but ACE2 and liver lipid droplet content are significantly correlated across the MASLD spectrum.Hepatic ACE2 levels are also increased in COVID-19 patients,especially those showing evidence of LI,but are not correlated with the presence of SARS-CoV-2 virus in the liver.However,there is a clear association between the hepatic lipid droplet content and the presence of the virus,suggesting a possible functional link.CONCLUSION Hepatic ACE2 levels were elevated in nonfibrotic MASH and COVID-19 patients with LI,while lipid accumulation may promote intra-hepatic SARS-CoV-2 replication,accelerating MASLD progression and COVID-19-mediated liver damage.

Timosaponin AⅢ induces drug-metabolizing enzymes by activating constitutive androstane receptor (CAR) via dephosphorylation of the EGFR signaling pathway

The current study aimed to assess the effect of timosaponin AⅢ(T-AⅢ)on drug-metabolizing enzymes during anticancer therapy.The in vivo experiments were conducted on nude and ICR mice.Following a 24-day administration of T-AⅢ,the nude mice exhibited an induction of CYP2B10,MDR1,and CYP3A11 expression in the liver tissues.In the ICR mice,the expression levels of CYP2B10 and MDR1 increased after a three-day T-AⅢ administration.The in vitro assessments with HepG2 cells revealed that T-AⅢ induced the expression of CYP2B6,MDR1,and CYP3A4,along with constitutive androstane receptor(CAR)activation.Treatment with CAR siRNA reversed the T-AⅢ-induced increases in CYP2B6 and CYP3A4 expression.Furthermore,other CAR target genes also showed a significant increase in the expression.The up-regulation of murine CAR was observed in the liver tissues of both nude and ICR mice.Subsequent findings demonstrated that T-AⅢ activated CAR by inhibiting ERK1/2 phosphorylation,with this effect being partially reversed by the ERK activator t-BHQ.Inhibition of the ERK1/2 signaling pathway was also observed in vivo.Additionally,T-AⅢ inhibited the phosphorylation of EGFR at Tyr1173 and Tyr845,and suppressed EGF-induced phosphorylation of EGFR,ERK,and CAR.In the nude mice,T-AⅢ also inhibited EGFR phosphorylation.These results collectively indicate that T-AⅢ is a novel CAR activator through inhibition of the EGFR pathway.

Changes in the Activities of Enzymes Involved in Starch Synthesis in the Kernel During Grain Filling in Winter Wheat Cultivars of Different Spike Types

Two winter wheat(Triticum aestivum L.)cultivars, large-spike type Yumai66 and small-spike type Yumai49, were used to study the activities of enzymes involved in starch synthesis in the kernel during grain filling. Starch accumulated faster in the kernel of Yumai49 than Yumai66 up to 25 d after anthesis, thereafter starch accumulated faster in the kernel of Yumai66. Starch accumulation in Yumai66 peaked at 20 -25 d after anthesis, while in Yumai49 starch accumulation peaked at 15 -20 d after anthesis and 25 -30 d after anthesis. The first peak was much higher than that of the second. Sucrose content and sucrose synthase activity peaked at 20 and 15 d after anthesis in Yumai66 and Yumai49, respectively. The sucrose content and sucrose synthase activity in Yumai66 were higher than that in Yumai49 during grain filling. ADP-glucose pyro-phosphorylase and starch branching enzyme activity in the kernel of Yumai66 peaked at 20 d after anthesis. while soluble starch synthase activity peaked at 10 and 20 d after anthesis. The second peak was much higher than that of the first.

Preparative Fractionation of Feruloyl Oligosaccharides Produced by Combinatorial Enzyme Digestion of Arabinoxylan

Pretreated wheat insoluble arabinoxylan was converted to oligosaccharides of structural variants using combinatorial enzyme approach. The digestive products were separated by preparative scale chromatographic Amberlite XAD-2 column. Fractions containing feruloyl oligosaccharides (FOS) were isolated, pooled, freeze-dried, and demonstrated to possess antimicrobial activity. The FOS suppressed cell growth of the test organism ATCC 8739 E. coli with a MIC value of 0.028% (w/v, 35˚C, 24 hr). The antimicrobial action was observed exceeding 72 hr of culture incubation. The FOS product could be a useful source of prebiotics or preservatives. The present results further confirm the science and application of the concept of combinatorial enzyme technique.

Effect of Soil Drought on C4 Photosynthetic Enzyme Activities of Flag Leaf and Ear in Wheat

The activities of RuBPC and C4 photosynthetic enzymes in ear and flag leaf blade were examined in wheat. The results showed that photosynthesis of ear was less sensitive to soil drought than that of flag leaf, and decrease of CO2 assimilation in flag leaf blade with water stress was more than that in ear. Compared with flag leaf, ear organs (awn, glume and lemma) had higher C4 enzyme activities and lower RuBPC activity. Under moderate water-stress, the increase of C4 enzyme activities was induced, and the increase was higher in ear than in flag leaf. Under severe water-stress, relatively higher C4 enzyme activities were still maintained in ear, rather than that in flag leaf. It suggests that high activities of C4 enzymes in ear may contribute to its high tolerance of photosynthesis to water-stress.

Physiological Effects of Chitosan Coating on Wheat Growth and Activities of Protective Enzyme with Drought Tolerance

Seedling period is an important stage of plant growth. This research was mainly to analysis the influence of chitosan on wheat seedling growth and physiological mechanisms under drought stress. The results showed that the group coated with chitosan significantly improved the growth index such as germination rate, wet weight, root length, root active, and impacted physiological indices such as superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT)), the content of malwondialdehyde (MDA) and chlorophyll compared with CK under drought stress. Activities of POD, CAT and SOD increased and then decreased, the content of MDA increased under drought stress. But. variation rates of the group coated with chitosan were slower than that of CK, which indicates that chitosan can significantly improve anti-oxidant enzymes activity to clear timely active oxygen and reduce the content of MDA so as to alleviate the degree of damage in the drought stress and make seedlings grow better. The results also showed that chitosan improved chloro-phyll content than that of CK, which demonstrated that chlorophyll content significantly influenced the photosynthetic efficiency of the mutant and added wheat above ground biomass and the field experiment results showed that chitosan increased yield 13.6% than that of CK.

Humic and Oxalic Acid Stimulates Grain Yield and Induces Accumulation of Plastidial Carbohydrate Metabolism Enzymes in Wheat Grown under Sandy Soil Conditions

Humic and oxalic acids have the effects of promoting plant growth. We test whether they are able to positively impact wheat yield under newly reclaimed sandy soil, where water deficiency negatively influences yield. Foliar application of humic acid and oxalic acid on two wheat cultivars, Gemiza-9 and Sakha-93, leads to overall better performance of the plants and increases the yield significantly, irrespective of the cultivar genetic background. However, Gemiza-9 surpassed Sak- ha-93 in grain yield parameters. The highest values of grain and protein yields/ha were obtained in both cultivars, when the plants were sprayed with a combination of 17 mg/L humic acid and (300 mg/L) oxalic acid. Humic and oxalic acid showed accumulative yield-promoting effect. To understand the mechanism by which humic and oxalic acids promoted grain yield, we performed SDS-PAGE followed by MS-MS-LC analyses. We identified a unique humic acid-induced 52 KDa band in Gemiza-9. The band contained three major proteins, Ribulose bisphosphate carboxylase large chain, ADP-glucose synthase and NADP-dependent glyceraldehyde-3-phosphate dehydrogenase (GAPN). Thus humic acid increased the activity of plastid enzymes involved in photosynthesis, sucrose biosynthesis and starched accumulation to improve the overall performance of the plant.

Alleviation of Adverse Effects of Salt Stress in Wheat Cultivars by Foliar Treatment with Antioxidant 2—Changes in Some Biochemical Aspects, Lipid Peroxidation, Antioxidant Enzymes and Amino Acid Contents

Ascobin (compound composed of ascorbic acid and citric acid) is considered one of exogenous protectants which may alleviate the harmful effects of salinity stress. Pot experiments were performed at the screen greenhouse of National Research Centre, Cairo, Egypt to study the effect of foliar treatment of two cultivars of wheat plant with different concentrations of ascobin (0, 200, 400 and 600 mg/l) on some biochemical parameters, antioxidant enzymes, element contents and amino acid constituents of two cultivars of wheat plant grown under different salinity levels (0.0, 3000 and 6000 mg/l) in 2011/2012 and 2012/2013. Salinity with different concentrations levels increased phenolic compounds contents of the two wheat cultivars. The activities of antioxidant enzymes (SOD, CAT, POD, PPO, AXP and GR) dramatically increased due to salinity stress. Amino acid content was increased in cultivar Sids 1, while the content was decreased in cultivar Giza 168 in all salinity treatments. Increments in the above mentioned parameters compared to the untreated plants at normal and stressed conditions. The magnitude of increments was much more pronounced in response to 600 mg/l of ascobin. It could be concluded that, foliar treatment of wheat cultivars with ascobin could partially alleviate the harmful effect of salinity especially at the lower levels of salinity of the two cultivars of wheat at most of the studied parameters.

Engineering high amylose and resistant starch in maize by CRISPR/Cas9-mediated editing of starch branching enzymes

To improve the amylose content(AC)and resistant starch content(RSC)of maize kernel starch,we employed the CRISPR/Cas9 system to create mutants of starch branching enzyme I(SBEI)and starch branching enzyme IIb(SBEIIb).A frameshift mutation in SBEI(E1,a nucleotide insertion in exon 6)led to plants with higher RSC(1.07%),lower hundred-kernel weight(HKW,24.71±0.14 g),and lower plant height(PH,218.50±9.42 cm)compared to the wild type(WT).Like the WT,E1 kernel starch had irregular,polygonal shapes with sharp edges.A frameshift mutation in SBEIIb(E2,a four-nucleotide deletion in exon 8)led to higher AC(53.48%)and higher RSC(26.93%)than that for the WT.E2 kernel starch was significantly different from the WT regarding granule morphology,chain length distribution pattern,X-ray diffraction pattern,and thermal characteristics;the starch granules were more irregular in shape and comprised typical B-type crystals.Mutating SBEI and SBEIIb(E12)had a synergistic effect on RSC,HKW,PH,starch properties,and starch biosynthesis-associated gene expression.SBEIIa,SS1,SSIIa,SSIIIa,and SSIIIb were upregulated in E12 endosperm compared to WT endosperm.This study lays the foundation for rapidly improving the starch properties of elite maize lines.

Wheat Straw Burial Improves Physiological Traits, Yield and Grain Quality of Rice by Regulating Antioxidant System and Nitrogen Assimilation Enzymes under Alternate Wetting and Drying Irrigation

Wheat straw burial has great potential to sustain rice production under alternate wetting and drying(AWD)irrigation.A field experiment was conducted with three wheat straw burial treatments,including without straw burial(NSB),with light straw burial of 300 kg/hm^(2)(LSB)and dense straw burial of 800 kg/hm^(2)(DSB),as well as three AWD regimes:alternate wetting/moderate drying(AWMD),alternate wetting/severe drying(AWSD)and alternate wetting/critical drying(AWCD).The rice growth and grain quality were higher in LSB and NSB than those in NSB under the same AWD regime.The AWMD×DSB treatment resulted in the highest yield,brown rice rate,milled rice rate,amylose content and protein content.Conversely,the AWCD×NSB treatment led to the lowest yield,brown rice rate,milled rice rate,amylose content and protein content.The active absorption area and nitrate reductase activity of roots were higher in the AWMD×DSB treatment than those in the AWCD×NSB treatment,as the former increased organic carbon and nitrogen contents in the rhizosphere,whereas the latter reduced their availability.Total soluble protein content and glutamine synthetase activity were greater in the AWMD×DSB treatment than those in the AWCD×NSB treatment.The activities of superoxide dismutase and catalase were higher in the AWMD×DSB treatment compared with the AWCD×NSB treatment,leading to the amelioration of oxidative cell injury,as shown by a lower malonaldehyde level.This study suggested that farmers should implement AWMD irrigation after leaving the straw residues in the field,followed by deep tillage to improve soil quality and mitigate the drought stress cycles of AWD.This approach can improve rice growth and grain quality and alleviate the problems of disposal of straw residues and water scarcity for sustainable rice production.

Addition of Protease Enzyme to Dog and Cat Feed and Its Influence on the Digestibility Coefficient, Immune Response, and Metabolic Biomarkers

Promoting better protein digestibility through exogenous enzymes in the diet is essential in the nutrition of companion animals, mainly for better performance and maintenance of the animals’ physiological and metabolic systems, promoting health and adequate growth. In addition, it reduces the cost of the diet by possibly reducing protein in the diet, which is the main and most expensive ingredient for dogs and cats. The objective of this study was to verify whether the addition of protease to dog and cat food can improve the protein digestibility of the food, thus facilitating greater absorption of amino acids and influencing metabolic biomarkers and immune response. To this end, two experiments were carried out to evaluate the protease from the fermentation of Aspergillus niger and Bacillus subtilis. Experiment 1 was carried out with ten male, non-castrated beagles divided into two groups of five animals: the control group (without enzyme) and the test group (with 250 g of protease/ton). The animals underwent two 45-day experimental periods, and in the second period, after a 15-day interval, the dogs in the control group became part of the treatment group (crossover model). Adding this enzyme to the dogs’ diet had no adverse effects on the animals’ health besides improving the digestibility of dry matter and crude protein consumed by the dogs. Experiment 2 was carried out with sixteen female cats of no defined breed, non-castrated, divided into four groups with four animals per group, namely: Treatment A (without enzyme), Treatment B (with protease at a dose of 100 g/ton), Treatment C (with protease at a dose of 200 g/ton) and Treatment D (with protease at a dose of 400 g/ton). The cats underwent two 30-day experimental periods, and in the second period, after a 15-day interval, the animals switched between treatments (crossover model) to increase the power of the statistical test. The enzyme consumption did not affect the felines’ metabolism and health but improved the digestibility of crude protein at doses of 200 and 400 g/ton. The results allow us to conclude that the protease used in this study can improve the digestibility of crude protein for dogs and cats.