Efficacy of incremental loads of cow's milk as a treatment for lactose malabsorption in Japan

BACKGROUND Lactose intolerance(LI)is commonly seen in East Asian countries.Several studies showed that lactose or milk loading has been used as a treatment for lactose malabsorption(LM)in Western countries,but there have been no reports regarding this type of treatment in Japan.As lactose or milk loading requires ingestion of large amounts of lactose within a short period,this is considered to be too harsh for Japanese people because of their less habitual milk consumption(175 mL per day in average)than Western people.In this study,we demonstrated lactose tolerance acquisition in a suitable way for Japanese.AIM To examine the efficacy of lactose(cow’s milk)loading treatment in patients with LM.METHODS Individuals with abdominal symptoms induced by milk or dairy products(LI symptoms)were identified with a questionnaire.A 20 g lactose hydrogen breath test(LHBT)was carried out to confirm LM diagnosis and to evaluate co-existence of small intestinal bacterial overgrowth(SIBO).Respondents diagnosed with LM were selected as study subjects and were treated with incremental loads of cow’s milk,starting from 30 mL and increasing up to 200 mL at 4-7 d intervals.After the treatment,changes in symptoms and LM diagnostic value of 20 g LHBT were investigated.Stool samples pre-and post-treatment were examined for changes in intestinal microbiota using 16S rRNA sequencing.Informed consent was obtained prior to each stage of the study.RESULTS In 46 subjects with LI symptoms(10-68 years old,mean age 34 years old)identified with the questionnaire,35(76.1%)were diagnosed with LM by 20 g LHBT,and 6 had co-existing SIBO.The treatment with incremental cow’s milk was carried out in 32 subjects diagnosed with LM(14-68 years old,median age 38.5 years old).The mean period of the treatment was 41±8.6 d.Improvement of symptoms was observed in 29(90.6%;95%confidence interval:75.0%-98.0%)subjects.Although 20 g LHBT indicated that 10(34.5%)subjects had improved diagnostic value of LM,no change was observed in 16(55.2%)subjects.Analysis of the fecal intestinal microbiota showed a significant increase in Blautia in 7 subjects who became symptom-free after the treatment(P=0.0313).CONCLUSION LM was diagnosed in approximately 75%of the subjects who had LI.Incremental loads of cow’s milk is regarded as a useful treatment for LM without affecting everyday life.

Hydrogen breath test for the diagnosis of lactose intolerance,is the routine sugar load the best one?

AIM: To evaluate the prevalence of lactose intolerance (LI) following a load of 12.5 g in patients diagnosed as high-grade malabsorbers using the hydrogen breath test (HBT)-25.METHODS: Ninety patients showing high-grade malabsorption at HBT-25 were submitted to a second HBT with a lactose load of 12.5 g.Peak hydrogen production,area under the curve of hydrogen excretion and occurrence of symptoms were recorded.RESULTS: Only 16 patients (17.77%) with positive HBT-25 proved positive at HBT-12.5.Hydrogen production was lower as compared to HBT-25 (peak value 21.55 parts per million (ppm) ± 29.54 SD vs 99.43 ppm ± 40.01 SD; P < 0.001).Symptoms were present in only 13 patients.The absence of symptoms during the high-dose test has a high negative predictive value (0.84) for a negative low-dose test.The presence of symptoms during the first test was not useful for predicting a positive low-dose test (positive predictive value 0.06-0.31).CONCLUSION: Most patients with a positive HBT-25 normally absorb a lower dose of lactose and a strict lactose restriction on the basis of a "standard" HBT is,in most instances,unnecessary.Thus,the 25 g lactose tolerance test should probably be substituted by the 12.5 g test in the diagnosis of LI,and in providing dietary guidelines to patients with suspected lactose malabsorption/intolerance.

The role of lactose in weanling pig nutrition:a literature and meta-analysis review

Lactose plays a crucial role in the growth performance of pigs at weaning because it is a palatable and easily digestible energy source that eases the transition from milk to solid feed.However,the digestibility of lactose declines after weaning due to a reduction in endogenous lactase activity in piglets.As a result,some lactose may be fermented in the gastrointestinal tract of pigs.Fermentation of lactose by intestinal microbiota yields lactic acid and volatile fatty acids,which may positively regulate the intestinal environment and microbiome,resulting in improved gastrointestinal health of weanling pigs.We hypothesize that the prebiotic effect of lactose may play a larger role in weanling pig nutrition as the global feed industry strives to reduce antibiotic usage and pharmacological levels of zinc oxide and supra-nutritional levels of copper.Evidence presented in this review indicates that high dietary lactose improves growth performance of piglets,as well as the growth of beneficial bacteria,particularly Lactobacillus,with the positive effects being more pronounced in the first 2 weeks after weaning.However,the risk of post-weaning diarrhea may increase as pigs get older due to reduced lactase activity,high dietary lactose concentrations,and larger feed intakes,all of which may lead to excessive lactose fermentation in the intestine of the pig.Therefore,dietary lactose levels exert different effects on growth performance and gastrointestinal physiological functions in different feeding phases of weanling pigs.However,no formal recommendation of lactose for weanling pigs has been reported.A meta-analysis approach was used to determine that diets fed to swine should include 20%,15%,and 0 lactose from d 0-7,d 7-14,and d 14-35 post-weaning,respectively.However,sustainable swine production demands that economics must also be taken into account as lactose and lactose containing ingredients are expensive.Therefore,alternatives to lactose,so called“lactose equivalents”have also been studied in an effort to decrease feed cost while maintaining piglet performance with lower dietary lactose inclusions.In summary,the present review investigated dose-response effects of dietary lactose supplementation to exert positive responses and begin to elucidate its mechanisms of action in postweaning pig diets.The results may help to replace some or all lactose in the diet of weanling pigs,while improving production economics given the high cost of lactose and availability in some swine production markets.

Differing Roles for Clostridium acetobutylicum’s Galactose Utilization Pathways

There has been a surge of interest in acetone-butanol-ethanol fermentations of Clostridium acetobutylicum due to its capacity to ferment many carbohydrates found in biomass. This metabolic diversity makes it a promising candidate for conversion of inexpensive, heterogeneous carbohydrate feedstocks to biofuels. Galactose is present in many such feedstocks due to its incorporation in plant cell walls. C. acetobutylicum encodes two galactose utilization pathways, the Leloir (LP) and the tagatose-6-P (T6P), and a previous study indicated genes for these pathways was differentially regulated during growth on galactose and lactose. In the current study we utilized quantitative PCR to further investigate gene expression levels and to show both pathways which were subject to carbon catabolite repression. During growth on galactose, mRNA for galactose-6-P isomerase from the T6P was induced to a greater extent than mRNA for glactokinase, the first enzyme in the LP. The galactose-6-P isomerase mRNAs were also more abundant than galactokinase mRNAs during growth on galactose. Analysis of theoretical ATP requirements to generate essential precursor metabolites indicated: 1) the LP is more efficient at generating upper glycolytic intermediates, 2) the T6P is more efficient at forming ATP, lower glycolytic intermediates and TCA cycle intermediates, 3) a combination of the two pathways is most efficient for forming precursor metabolites found in the pentose phosphate pathway. From this it can be suggested that the two pathways have different roles in the organism with the T6P generating most ATP and precursor metabolites and the LP providing upper glycolytic metabolites.

Management and treatment of lactose malabsorption

Lactose malabsorption is a very common condition characterized by intestinal lactase deficiency. Primary lactose malabsorption is an inherited deficit present in the majority of the world＇s population, while secondary hypolactasia can be the consequence of an intestinal disease. The presence of malabsorbed lactose in the colonic lumen causes gastrointestinal symptoms. The condition is known as lactose intolerance. In patients with lactase nonpersistence, treatment should be considered exclusively if intolerance symptoms are present. In the absence of guidelines, the common therapeutic approach tends to exclude milk and dairy products from the diet. However, this strategy may have serious nutritional disadvantages. Several studies have been carried out to find alternative approaches, such as exogenous β-galactosidase, yogurt and probiotics for their bacterial lactase activity, pharmacological and non pharmacological strategies that can prolong contact time between enzyme and substrate delaying gastrointestinal transit time, and chronic lactose ingestion to enhance colonic adaptation. In this review the usefulness of these approaches is discussed and a therapeutic management with a flow chart is proposed.

Four-sample lactose hydrogen breath test for diagnosis of lactose malabsorption in irritable bowel syndrome patients with diarrhea

AIM: To validate 4-sample lactose hydrogen breath testing(4SLHBT) compared to standard 13-sample LHBT in the clinical setting.METHODS: Irritable bowel syndrome patients with diarrhea(IBS-D) and healthy volunteers(HVs) were enrolled and received a 10 g, 20 g, or 40 g doselactose hydrogen breath test(LHBT) in a randomized, double-blinded, controlled trial. The lactase gene promoter region was sequenced. Breath samples and symptoms were acquired at baseline and every 15 min for 3 h(13 measurements). The detection rates of lactose malabsorption(LM) and lactose intolerance(LI) for a 4SLHBT that acquired four measurements at 0, 90, 120, and 180 min from the same data set were compared with the results of standard LHBT.RESULTS: Sixty IBS-D patients and 60 HVs were studied. The genotype in all participants was C/C-13910. LM and LI detection rates increased with lactose dose from 10 g, 20 g to 40 g in both groups(P < 0.001). 4SLHBT showed excellent diagnostic concordance with standard LHBT(97%-100%, Kappa 0.815-0.942) with high sensitivity(90%-100%) and specificity(100%) at all three lactose doses in both groups.CONCLUSION: Reducing the number of measurements from 13 to 4 samples did not significantly impact on the accuracy of LHBT in health and IBS-D. 4SLHBT is a valid test for assessment of LM and LI in clinical practice.

Optimization of Cultural Condition and Synergistic Effect of Lactose with Carboxymethyl Cellulose on Cellulase Production by <i>Bacillus</i>sp. Isolated from Fecal Matter of Elephant (<i>Elephas maximus</i>)

A cellulase producing bacterium (E3 strain) was isolated from fecal matter of elephant and identified as Bacillus sp. using 16S rDNA sequenced based molecular phylogenetic approach. While studying the effect of substrates like Carboxymethyl cellulose (CMC), avicel, starch, maltose, sucrose, glucose, fructose, galactose and lactose on cellulase production, it was found that CMC was best carbon source induced cellulase production followed by lactose in this bacterial strain. A positive synergistic effect of lactose with CMC was also observed with enhancement of 5 - 6 times in cellulase production. The optimum cellulase production was recorded with 1% CMC and 1% lactose when added individually in the Omeliansky’s medium. The results showed that addition lactose with CMC greatly enhances the production and activity of various cellulase enzymes. The optimal fermentation conditions for the biosynthesis of cellulase by this strain were found to be temperature: 37℃, pH 7.0. The nitrogen source NH4Cl at 0.15% was optimum for cellulase production by this bacterium.

Lactose malabsorption and intolerance: What should be the best clinical management?

Lactose malabsorption (LM) is the incomplete hydrolysis of lactose due to lactase deficiency, which may occur as a primary disorder or secondary to other intestinal diseases. Primary adult-type hypolactasia is an autosomal recessive condition resulting from the physiological decline of lactase activity. Different methods have been used to diagnose LM. Lactose breath test represents the most reliable technique. A recent consensus conference has proposed the more physiological dosage of 25 g of lactose and a standardized procedure for breath testing. Recently a new genetic test, based on C/T13910 polymorphism, has been proposed for the diagnosis of adult-type hypolactasia, complementing the role of breath testing. LM represents a wellknown cause of abdominal symptoms although only some lactose malabsorbers are also intolerants. Diagnosing lactose intolerance is not straightforward. Many non-malabsorber subjects diagnose themselves as being lactose intolerant. Blind lactose challenge studies should be recommended to obtain objective results. Besides several studies indicate that subjects with lactose intolerance can ingest up to 15 g of lactose with no or minor symptoms. Therefore a therapeutic strategy consists of a lactose restricted diet avoiding the nutritional disadvantages of reduced calcium and vitamin intake. Various pharmacological options are also available. Unfortunately there is insufficient evidence that these therapies are effective. Further double-blind studies are needed to demonstrate treatment effectiveness in lactose intolerance.

Preparation of slab-shaped lactose carrier particles for dry powder inhalers by air jet milling

Dry powder inhalers are often formulated by attaching micronized drug particles onto carrier particles, which are generally lactose. In this study, commercially available lactose was air jet milled to produce unique slab-like coarse carrier particles, which have larger and rougher surfaces compared to other commercially available lactose. Two key processing factors, i.e.,classifier speed and jet milling pressure, were systematically investigated. The largest fraction of slab-like particles in the resulting powder was obtained at a classifier speed of 3000 rpm.The slab-like coarse carrier particles are expected to exhibit superior performance than commercial lactose due to their unique surface properties.

Enzymatic Regioselective Synthesis of Vinyl Lactose Ester and Its Chemical Polymerization

Transesterification reaction of lactose with divinyladipate in pyridine catalyzed by an alkaline protease from Bacillus subtilis at 50C for 3 days gave 6-O-vinyladipoyl-lactose (yield 35%). Poly(6-O-vinyladipoyl-lactose) with Mn = 21,200, Mw = 32,900, Mw/Mn = 1.56 could be obtained by chemical polymerization. Poly(vinyl alcohol) containing lactose branch was biodegradable. After 6 days in aqueous buffer (PH 7), this alkaline protease could degrade the polymer to an Mn of ca. 2100, Mw/Mn = 2.56.

Studies on the spray dried lactose as carrier for dry powder inhalation

The purpose of this study was to investigate the spray dried lactose as carrier for dry powder inhalation(DPI).The lactose particles were prepared by spray drying,then the particle size,shape and crystal form were characterized by laser diffraction,scanning electron microscopy(SEM),X-ray diffraction(XRD)and differential scanning calorimetry(DSC).The spray dried lactose particles were spherical and amorphous,but would transfer to crystal form when storage humidity was above 32%.Thus,the humidity of the storage environment should be controlled below 30%strictly in order to maintain the amorphous nature of spray dried lactose which is a great benefit to DPI development.

Milk Consumption and Lactose Intolerance in Adults

Objective To investigate relations between milk consumption and lactose intolerance （LI） in adults and to explore the effect of milk consumption on lactase activity.Methods Total of 182 subjects aged 20‐70 years were recruited and interviewed by questionnaires,and their accumulative cow＇s milk intake （AMI） was calculated.LI was evaluated by hydrogen breath test （HBT）.Results A negative correlation was found between AMI and severity of observed LI symptom （r=-0.2884;P0.05）.Binary logistic regression analysis showed a negative correlation between LI and duration and frequency of milk consumption （OR,0.317 and 0.465,respectively;both P0.05） and a positive correlation between LI and amount of milk consumed per sitting （OR,6.337;P0.05）.Conclusion LI is related to various milk consumption behaviors.Most Chinese adults with LI may tolerate moderate milk consumption 160 mL.

Hydrolysis of Lactose： Estimation of Kinetic Parameters Using Artificial Neural Networks

The analysis of any kinetic process involves the development of a mathematical model with predictive purposes. Generally, those models have characteristic parameters that should be estimated experimentally. A typical example is Michaelis-Menten model for enzymatic hydrolysis. Even though conventional kinetic models are very useful, they are only valid under certain experimental conditions. Besides, frequently large standard errors of estimated parameters are found due to the error of experimental determinations and/or insufficient number of assays. In this work, we developed an artificial neural network （ANN） to predict the performance of enzyme reactors at various operational conditions. The net was trained with experimental data obtained under different hydrolysis conditions of lactose solutions or cheese whey and different initial concentrations of enzymes or substrates. In all the experiments, commercial 13-galactosidase either free or immobilized in a chitosan support was used. The neural network developed in this study had an average absolute relative error of less than 5% even using few experimental data, which suggests that this tool provides an accurate prediction method for lactose hydrolysis.

Multi-Drug Resistance Pattern of Lactose Non-Fermenting <i>Escherichia coli</i>as Causative Agent of Urine Tract Infections in Luanda, Angola

This prospective study was carried out to assess the sensitivity and resistance pattern of lactose non-fermenting Escherichia coli from July 2018 to December 2018 in the Laboratory of Microbiology at Luanda Medical Center, Angola. Out of 1170 patient, a total of 120 urine specimens infected with Escherichia coli (>105 CFU/ml) were collected according to the routine protocol of urinalysis. Among these 120 isolates, 25 (21%) isolates were determined as “atypical”, lactose non-fermenting E. colis trains. The twenty-five lactose non-fermenting Escherichia coli strains isolated from urine samples in Luanda Medical Center were declared as Multiple Drugs-Resistant strains with high resistance to Cefalexine (100%), Cefuroxime (100%), Ceftriaxone (92%), Gentamycin (92%), Ciprofloxacin (72%) and Amoxiciclin/Clavulanic (80%). The alarming resistance level to the first-choice drugs for the treatment of urinary tract infections caused by non-fermentative lactose E. coli was observed.

Isolation, Characterization and Antimicrobial Resistance Patterns of Lactose-Fermenter Enterobacteriaceae Isolates from Clinical and Environmental Samples

The lactose-fermenter Enterobacteriaceae are the most frequent cause of clinical infection in our country. The objective of this study was to isolate and identify the most common lactose-fermenter Enterobacteriaceae from clinical samples, including urine, blood, wounds, and sputum, obtained from the local hospital and from environmental samples from a chicken farm, agriculture soil, and water from the Tigris River in Baghdad City. The study also aimed at establishing the antibiotic resistance patterns of the isolated bacteria. A total of 155 bacterial isolates were identified from 10 genera according to the Vitek 2 system. The most common bacterial isolates from the clinical and environmental samples were Escherichia coli and Klebsiella pneumoniae, respectively. The antibiotic resistance patterns showed that all clinical and environmental isolates were multidrug resistant to β-lactam (except carbapenems) drug and aminoglycosides and more sensitive to carbapenems.

Effect of Temperature, pH and Amount of Enzyme Used in the Lactose Hydrolysis of Milk

Lactose intolerance is becoming a health state that requires the restriction of dairy products in the diet of people suffering from this condition. But milk and dairy products, due to a well-balanced composition in the main macro and micronutrients, cannot be missing from the diet of the consumer of any age. For these reasons, in recent years, in the milk processing industry, the production of low-lactose or lactose-free dairy products is explored. To reduce the lactose content of dairy raw materials, various industrial and biotechnological methods were used: enzymatic hydrolysis of lactose, baromembranous methods, bioconversion of lactose by lactic bacteria and others. The most widely used lactase enzymes in the industry are mesophilic enzymes from filamentous fungi (Aspergillus spp.) and yeasts (Kluyveromyces spp.). Therefore, the aim of this study was to evaluate the effect of the commercial enzyme β-galactosidase on the hydrolysis of cow’s milk at different enzyme concentrations, temperatures and pH. Two commercial enzymes β-galactosidase obtained from Bacillus licheniformis and β-galactosidase obtained from Kluyveromyces lactis, were used in this study, according to information provided by the manufacturer. The thermal stability of lactose, the effect of milk pH, the effect of temperature, duration of hydrolysis and the amount of enzymes on the lactose hydrolysis degree and the sweetness degree of milk were determined. Research has identified the optimal parameters for obtaining a high degree of lactose hydrolysis in the use of these enzymes. Therefore, to ensure a high lactose hydrolysis degree (over 80%), the following lactose hydrolysis regimens were identified: temperature 4°C - 6°C, 0.3% Bacillus licheniformis enzymes, duration 4 hours;temperature 4°C - 6°C, 0.3% enzymes from Klavyromyces lactis, duration 12 hours and temperature 38°C - 40°C, 0.15% enzymes from (Bacillus licheniformis or Klavyromyces lactis), duration 2 - 3 hours. The results obtained allow the efficient use of Bacillus licheniformis and Klavyromyces lactis enzymes in industrial processes for the manufacture of “lactose-free” or “low-lactose” drinking milk and fermented dairy products for people with lactose intolerance.

Osteryoung square wave voltammetric determination of lactose in food samples by a derivative procedure

A method for determination of lactose in food samples by Osteryoung square wave voltammetry （OSWV） was developed. It was based on the nucleophilic addition reaction between lactose and aqua ammonia. The carbonyl group of lactose can be changed into imido group, and this increases the electrochemical activity in reduction and the sensitivity. The optimal condition for the nucleophilic addition reaction was investigated and it was found that in NH4C1-NH3 buffer of pH 10.1, the linear range between the peak current and the concentration of lactose was 0.6-8.4 mg L-l, and the detection limits was 0.44 mg L- 1. The proposed method was applied to the ＇determination of lactose in food samples and satisfactory results were obtained.

Physico-Chemical, Microbiological and Sensory Characteristics of Yogurt as Affected by Added Lactose

Enrichment of yogurt with lactose addition may increase the growth of the yogurt starter culture (Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus) and enhance yogurt physico-chemical and sensory attributes. The objectives of this study were to determine the influence of added lactose on the 1) physico-chemical characteristics, including the final lactose content of yogurt, during its shelf life;2) growth of the yogurt starter culture over yogurt shelf life and 3) the sensory attributes of yogurt. Fat free plain set-type yogurt was manufactured using 0%, 1%, 3% and 5% w/w added lactose to accomplish objectives 1 and 2. For objective 3, a blueberry yogurt was manufactured using the same lactose levels. Analyses for plain set-type yogurt were conducted at 7 days intervals during 35 days of storage. Sensory evaluation was conducted on flavored yogurt three days after its manufacture. Data were analyzed using Proc Mixed model of SAS&reg;9.3 program. Significant differences between means were analyzed at α = 0.05 using Tukey adjustment. Lactose addition influenced some of the yogurt characteristics in a positive manner. Lactose contents of yogurts with lactose added at 1%, 3% and 5% stayed higher in that proportion than control throughout the 35 days of storage. Yogurts containing 5% w/w added lactose had the lowest pH. Yogurts containing 5% w/w added lactose had significantly the highest syneresis values compared to 0%, 1% and 3% w/w added lactose during storage period at day 7 and from day 21 onwards. Use of 5% w/w added lactose resulted in significantly higher counts of Streptococcus thermophilus compared to control and this bacterial survival was the highest for 1% w/w added lactose compared to the rest. The overall liking scores indicated that yogurts containing added lactose were preferred over control. For taste, sourness and sweetness samples containing added lactose had higher scores than control. The consumer acceptability of yogurts increased as lactose addition increased. The acceptability of yogurts and purchase intent frequency scores markedly increased with the addition of lactose.

Antilisterial Activity of Lactose Fatty Acid Esters in Milk

Carbohydrate fatty acid esters have been shown to inhibit the growth of bacteria in growth media. In this study, the antilisterial activities of the carbohydrate esters lactose monodecanoate (LMD) and lactose monolaurate (LML) were tested in milk samples with variable fat content (up to 10%) and at different temperatures (37&deg;C, 24&deg;C and 5&deg;C) after inoculation with a 5-strain Listeria monocytogenes cocktail. Both LMD and LML demonstrated antilisterial properties against L. monocytogenes in milk, with the observed log reductions in the LMD milk samples being higher than those in the LML samples at all tested temperatures. LMD in 1% and 2% fat milk was found to be bactericidal with no Listeria growth by day 6. LMD showed significant antilisterial activity in milk samples containing up to 9% fat at 37&deg;and in samples with up to 3.5% fat at 24&deg;C and in milk samples with up to 2% fat at 5&deg;C. LML showed significant Listeria log reductions at 37&deg;C with milk containing up to 9% fat but showed no microbial inhibition in milk in any samples at 24&deg;C and 5&deg;C. Although carbohydrate fatty acid esters have proven microbial bactericidal/bacteriostatic properties in growth media, their antimicrobial properties are reduced in a food system containing fat and at temperatures lower than 37&deg;C. The significance of this research is identifying food constituents and storage temperatures that limit the microbial inhibitory properties of lactose esters to ensure appropriate use as potential food grade antilisterial agents.

Emulsification Properties of Lactose Fatty Acid Esters

Sugar fatty acid esters (SFAE) are a class of synthetic emulsifiers used in the food, pharmaceutical, and personal care industries. The influence of the fatty acid chain length on the emulsification properties of lactose fatty acid esters (LFAE) including lactose monooctanoate (LMO), lactose monodecanoate (LMD), lactose monolaurate (LML) and lactose monomyristate (LMM) was investigated in this study. The stability of the emulsions as well as the oil droplet size distribution in 20% soybean oil-in-water emulsions was measured at 0.1%, 0.25% and 0.5% of LFAE concentrations. In order of LFAE with the strongest emulsion stabilization characteristics were LML, LMD, LMO and LMM. Oil droplet distributions resulted in the same trend, with LML and LMD maintaining the smallest droplet sizes and thus the most stable emulsion. The hydrophilic-lipophilic balance (HLB) and critical micelle concentrations were determined for each LFAE. An increase in HLB value was seen with an increased CMC value for each LFAE, showing the strength of the linear relationship between these two measured values. Additionally, there was a decrease in HLB and CMC values with a decrease in the fatty acid chain length of each LFAE. This research showed that LML and LMD formed more stable emulsions, even with HLB and CMC values higher than those of LMM suggesting HLB and CMC values alone do not predict emulsifier effectiveness.