Uncovering the Functional Potential of Goat Milk: Physicochemical and Rheological Comparison Across Local Goats in Gorontalo Indonesia

Uncovering the Functional Potential of Goat Milk: Physicochemical and Rheological Comparison Across Local Goats in Gorontalo Indonesia | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Uncovering the Functional Potential of Goat Milk: Physicochemical and Rheological Comparison Across Local Goats in Gorontalo Indonesia Agus Bahar Rachman, Fahrul Ilham, Lukman Hakim, Nicolays Jambang, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6429174/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 11 You are reading this latest preprint version Abstract The physicochemical and rheological characteristics of goat milk are pivotal in determining its processing potential and nutritional value. Despite Indonesia’s rich diversity in goat breeds, scientific data comparing the milk quality across these breeds remain scarce. This study investigated the physicochemical, rheological, structural, and protein characteristics of raw goat milk from four genotypes—Kacang, Local Goat, Etawa Crossbreed, and Saanen Crossbreed—reared under traditional farming conditions in Gorontalo, Indonesia. The aim was to identify breed-specific differences that may influence milk’s nutritional quality and processing suitability. A total of 48 raw milk samples were analyzed using standard compositional assays, sodium dodecyl sulfate–polyacrylamide gel electrophoresis, densitometry, amino acid profiling, laser confocal microscopy, rotational rheometry, and colloidal stability measurements. These methods provided detailed insights into protein structure, micelle behavior, and textural properties. The results revealed that crossbred goats, particularly Saanen and Etawa, produced milk with higher protein content, stronger casein expression, greater viscosity, and richer profiles of glutamic acid, proline, and threonine. Conversely, Kacang goats showed higher whey protein content and elevated sulfur amino acids, such as cysteine and methionine. All samples exhibited pseudoplastic flow behavior and comparable zeta potential and particle size, indicating similar colloidal stability. These findings demonstrate the influence of genotype on goat milk functionality and support its valorization in developing sustainable and differentiated dairy products with enhanced nutritional and technological properties. . Physicochemical Rheological Milk Local Goats Gorontalo Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Introduction Dairy goat farming in Indonesia has emerged as a promising sector, evidenced by a notable increase in the goat population of 2.89% in 2021, bringing the total to 19.23 million goats [ 1 ]. This upward trend reflects a growing awareness of the many benefits associated with goat milk, which is gaining recognition for its superior nutritional qualities [ 2 ]. Research indicates that goat milk offers a more favorable nutritional profile compared to cow milk, primarily due to its smaller fat globules. This unique characteristic not only enhances its creamy texture but also facilitates easier digestion [ 3 ]. Goat milk also has a higher content of conjugated linoleic acid and medium-chain fatty acids, which add to its unique flavor and certain health advantages [ 4 ]. Its impressive protein content, along with its buffering capacity and alkalinity, makes it particularly beneficial for individuals seeking to manage stomach ulcers [ 5 ] In addition, goat milk's peculiar casein micelle structure is less solvated and more heat stable than cow milk's, which is very beneficial in the dairy industry, especially when manufacturing cheese. [ 6 ]. Overall, the growth of the dairy goat industry in Indonesia holds significant promise for both producers and consumers as the appreciation for goat milk continues to grow [ 1 ]. Indonesia is renowned for its diverse dairy goat breeds, each playing an integral role in providing high-quality, nutritious milk and dairy products to local communities [ 7 ]. Among these breeds are the indigenous Kacang and notable crossbreeds such as the Saanen Crossbreed, Etawah Crossbreed, and Saanen-Etawah (Sapera). These breeds have been carefully developed through strategic crossbreeding with varieties from South Asia, Africa, and Europe [ 8 ]. The Indonesian government’s official recognition of the Etawah Crossbreed as a valuable local livestock breed highlights its commitment to preserving the nation’s agricultural heritage and fosters confidence in the future of the dairy industry [ 9 ]. Additionally, Saanen goat farms represent a highly effective alternative source of milk production, further enriching the local dairy sector [ 10 ]. In Gorontalo, innovative crossbreeding efforts between the Etawah Crossbreed and Kacang goats have resulted in the development of Local Gorontalo goats, which have successfully adapted to their local environment [ 8 ]. It is also important to emphasize the Kacang goat, an indigenous breed with significant untapped potential, which can greatly enhance dairy production in Indonesia [ 11 ]. The expansion and sustainability of the dairy sector in the area will be greatly aided by identifying and fostering these capabilities. Despite the huge goat population in Gorontalo, there is a lot of room to grow the usage of goat milk as a major source of animal protein. This underutilization highlights an opportunity for growth, particularly as consumer preferences favor cow's milk, often due to its more familiar flavor compared to the distinctive taste of goat milk. Several prominent goat breeds, including Kacang, Local Goat, Saanen Crossbreed, and Etawa Crossbreed, serve a dual purpose by providing both milk and meat [ 8 ] [ 12 ]. By prioritizing high-yield and productive breeds, breeders can make strategic decisions that align with the economic goals of goat farming [ 13 ]. Moreover, the increasing demand for goat milk with desirable functional properties reflects a promising market opportunity. By capitalizing on this potential, the community could significantly boost the consumption of goat milk [ 14 ]. Establishing a Transdisciplinary Community of Practice (TDCoP) that blends scientific research, local agricultural expertise, and industry needs could facilitate this advancement [ 15 ]. This collaborative approach would not only promote the sustainability of local agriculture but also enhance knowledge transformation in rural areas, benefiting both producers and consumers in Gorontalo [ 16 ]. This study aims to analyze the physicochemical composition, rheology, and microstructure of raw milk from four distinct goat breeds in Indonesia. It is anticipated that there will be significant variations in the chemical and physical compositions also rheology of the milk samples due to the influence of breed, which is a crucial determinant of milk composition. These insights are essential for goat breeders and milk product manufacturers as they can enhance breed selection, cross-breeding programs, and the development of nutritionally superior functional foods. Additionally, it's important to note that other factors like feed, environmental conditions, and daily care practices may also impact the characteristics of these milk samples. Materials and methods 2.1 Materials Fresh goat milk samples were gathered from 48 animals representing four different breeds: Kacang, Local Gorontalo, Saanen crossbreed, and Etawa crossbreed, with 12 animals from each breed. These animals were in the 8 months of lactation stage and were at a farm in Tulabolo Village, Suwawa District, Bone Bolango Regency, Province of Gorontalo, Indonesia. Their diet consisted of sorghum stem and leaves, dairy goat pellets, and corn After being manually extracted in the morning and aseptically stored in sterile glass bottles, the milk was brought to the lab in an icebox that was kept between 2 and 4°C. Subsequently, the samples were preserved at − 20°C for further use. 2.2 Physicochemical Composition of Goat Milk Using accepted practices, the samples were subjected to a comprehensive proximate composition analysis. AOAC Official Method 925 was used to compute the total solid content. 23 Milk Solids (Total) [ 17 ]. The protein content were measured using the Kjeldahl method (EN ISO 8968-1:2002)[ 18 ], while the fat content was accurately calculated using the Gerber method (AOAC 2000.18)[ 19 ]. The specific gravity were measured using a Lactodensimeter 66110 (Funke Gerber, Germany). Furthermore, pH measurements were conducted using a standardized pH level (model Eutech™ PC 700 Multi-parameter Meter, manufactured by Thermo Fisher Scientific, Waltham, MA, USA). with pH 4 and 7 buffer solutions. 2.3 Amino Acid Analysis by HPLC The analysis followed the procedures outlined with some modifications[ 20 ]. The thawed milk samples A 0.2 µm cellulose nitrate membrane filter was used to filter the resultant hydrolysate, and phenylisothiocyanate was used for pre-column derivatization. A reversed-phase JASCO HPLC System (JASCO, Tokyo, Japan) fitted with a JASCO Unifinepak C18 (3.0 mm I.D. x 75 mm L, 1.9 µm) was used to perform the amino acid analysis. (JASCO, Tokyo, Japan HPLC-grade acetonitrile containing 0.1 M ammonium acetate and 0.1 M ammonium acetate at pH 6.5: They used methanol: water (46: 1 0: 44).. The flow rate was set at 1.0 mL/min, and the response of the UV detector was seen at 254 nm. 2.4 Gel Electrophoresis Analysis The milk samples underwent a 10-minute, 12,000-g centrifugation at -4°C Next, the layer of fat was gently removed. After being collected from beneath the creamy layer, the milk serum was utilized for additional protein characterization procedures..The protein levels in milk samples were calculated using Lowry [ 21 ]. Then, the protein samples were solubilized with phosphate buffer at specified concentrations and a temperature of 20°C. Subsequently, the solubilized samples underwent sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using a Bio-Rad System (Mini-Protean 3 cell, USA) following the method described by Laemmli [ 22 ]. Following electrophoresis, the gel was stained with Coomassie blue dye for two hours and then destained in a solution containing 30% methanol and 10% acetic acid. The molecular weight marker 10–200 kDa (#SM0661) was from Fermentas (USA), and other reagents were of analytical grade. The Image Lab (Bio-Rad) software was used to examine the documentation's results using the densitometry technique. The software could predict molecular weight and relative density of each band which was relatively compared to bands from protein ladders/markers, so each molecular weight was determined. By calculating the standard protein quantity based on their density, the bands from BSA at varying concentrations were utilized to forecast the amount of each protein component. [ 23 ]. Because the software can automatically create a regression from the standard band data, the quantity of the target band could be predicted. The collected data was then statistically examined to compare each component of goat and cow milk using the Mann-Whitney test. 2.5 Particle Size and Zeta Potential Analysis The zeta potential and average particle size of raw goat milk were measured by diluting a 200 µL sample with 10 mL of 0.1 mol/L Na 2 HPO 4 -NaH 2 PO 4 (pH 7.0) buffer solution. To get rid of any dust and contaminants, the samples were then centrifuged at 1,078 × g for five minutes at room temperature. Then, using the technique described by Li et al the protein particle size was examined using a HORIBA NANO PARTICA SZ-100 (Kyoto, Japan) nanometer particle size analyzer [ 24 ]. A measurement temperature of 25°C, a scattering angle of 90°, a particle refractive index of 1.450, a particle absorptivity of 0.8872, and the usage of water as a dispersion with a dispersant refractive index of 1.330 were among the parameters considered in the analysis. Additionally, using a HORIBA NANO PARTICA SZ-100 (Kyoto, Japan) nanometer particle size analyzer with laser Doppler microelectrophoresis, the zeta potential of the goat milk samples was ascertained using the technique outlined by O'Brien et al [ 25 ]. 2.6 Color Analysis Color was measure on goat milk samples using the Chroma Meter CR-400 (Konica Minolta, Japan). The color readings were determined based on L* (Lightness), where L = 100 for white and L = 0 for black, Chroma a* (ranging from − 60 for green chromaticity to + 60 for red), and Chroma b* (ranging from − 60 for blue chromaticity to + 60 for yellow). It's important to note that the chroma meter was calibrated using a calibration plate. 2.7 Determination of Rheological Properties A slightly modified Rheometer (MCR302, ANTON PAAR GMBH, Graz, Austria) was used to measure the rheological characteristics of goat milk. The sample tank was filled with 25 milliliters of goat's milk. A CC25 DIN/TI rotor and a CCB25 DIN/SS cup test head with a 120.00 mm diameter were used in the test. A scanning shear rate of 0.1 to 500 s-1 was used, and the clearance was fixed at 1.00 mm. Shear stress and apparent viscosity measurements were recorded for the entire test, which was carried out at 25°C. The consistency coefficient (K) and flow behavior index (n) were then measured by fitting the shear stress and shear rate curves. 2.8 Confocal Microscopy Measurement Following Li and Shah's suggested methodology, the milk samples' structures were examined using the FV1200 confocal scanning laser microscope (Olympus) [ 26 ]. The confocal scanning laser microscopy was outfitted with a silicon oil objective lens and an inverted microscope (magnification 150×). A tagged image file format is used to acquire digital image files. Ten microliters of Nile red (1 mg/mL) were used to stain one milliliter of goat milk samples. in ethanol solution) and 10 µL of fluorescein isothiocyanate (FITC, 1 mg/mL in ethanol solution) for 30 min. Then, 20 µL of the stained material was pipetted onto a glass slide, covered with a coverslip, and put right into the confocal scanning laser microscope. In order to conduct the observations in a dark room, the emission wavelengths for FITC and Nile Red were set at 495 to 559 nm and 534 to 488 nm, respectively, and the excitation wavelengths were set at 500 to 600 nm. 2.9 Statistical Analysis Analysis of differences in milk quality between goat breeds was carried out using descriptive tests and one-way ANOVA using SPSS 23.0 software. Descriptive analysis includes the simplification and presentation of data so that the data that has been obtained in the field can provide preliminary information even though the information has not reached the stage of drawing conclusions. ANOVA was conducted to determine whether there was a difference in the diversity of the chemical quality of milk between the four goat breeds obtained. Using 30 variables chosen from Table 1 , Table 2 , and four examples of goat breeds, principal component analysis (PCA) was used to separate groups belonging to various breeds. Table 1 The chemical quality of goat milk Milk Quality Paramater Kacang Local Goat Etawa Crossbreed Saanen Crossbreed Moisture Content (%) 85.48 ± 2.14 c 86.86 ± 0.39 b 87.76 ± 1.84 a 87.52 ± 0.98 a Lactose (%) 4.55 ± 0.45 a 4.26 ± 0.32 a 4.38 ± 0.51 a 4.44 ± 0.26 a Fat (%) 4.08 ± 0.21 a 4.10 ± 0.13 a 4.09 ± 0.08 a 4.12 ± 0.28 a Protein (%) 3.75 ± 0.63 b 3.95 ± 0.19 b 4.11 ± 0.04 a 4.23 ± 0.02 a Amino Acids (g/ 100 g of Protein) Asp 0.29 ± 0.25 c 0.58 ± 0.11 c 3.14 ± 0.12 a 2.15 ± 0.23 b Glu 3.05 ± 0.12 c 5.31 ± 0.48 b 7.02 ± 0.32 a 7.14 ± 0.22 a Ser 1.43 ± 0.06 b 1.51 ± 0.25 b 2.28 ± 0.04 a 2.41 ± 0.25 a Gly 1.04 ± 0.10 b 1.39 ± 0.35 b 2.15 ± 0.42 a 2.05 ± 0.02 a His 0.34 ± 0.06 a 0.50 ± 0.01 a 0.92 ± 0.26 a 0.62 ± 0.00 a Arg 0.51 ± 0.13 c 0.78 ± 0.32 c 1.31 ± 0.22 b 2.18 ± 0.42 a Thr 2.87 ± 0.76 d 3.05 ± 0.42 c 6.24 ± 0.45 a 4.11 ± 0.03 b Pro 2.04 ± 0.02 c 2.56 ± 0.46 c 5.02 ± 0.18 a 3.11 ± 0.19 b Tyr 0.43 ± 0.01 b 0.62 ± 0.02 b 1.81 ± 0.09 a 1.21 ± 0.04 a Val 1.76 ± 0.14 c 1.95 ± 0.03 c 3.14 ± 0.08 a 2.29 ± 0.05 b Cys 0.05 ± 0.01 a 0.08 ± 0.05 a 0.10 ± 0.02 a 0.19 ± 0.01 a Met 0.32 ± 0.02 b 0.94 ± 0.06 b 1.25 ± 0.01 a 1.18 ± 0.01 a Ile 1.14 ± 0.02 b 1.21 ± 0.02 b 2.12 ± 0.13 a 1.97 ± 0.02 b Leu 1.28 ± 0.21 b 1.55 ± 0.03 b 2.44 ± 0.08 a 2.32 ± 0.02 a Phe 1.03 ± 0.11 a 1.13 ± 0.04 a 2.60 ± 0.05 a 1.94 ± 0.03 b Lys 1.35 ± 0.05 a 1.41 ± 0.01 a 1.92 ± 0.09 a 1.77 ± 0.01 a Notes: Different superscripts on the same line indicate significant differences (P < 0.05). Table 2 The physical quality of goat milk Milk Quality Paramater Kacang Local Goat Etawa Crossbreed Saanen Crossbreed Specific gravity 1.03320 ± 0.11 a 1.03540 ± 0.09 a 1.03340 ± 0.04 a 1.03450 ± 0.55 a Zeta Potential (mV) -8.48 ± 0.22 a -8.55 ± 0.17 a -8.65 ± 0.33 a -8.57 ± 0.45 a Particle Size (nm) 247 ± 0.74 a 244 ± 0.14 a 246 ± 0.32 a 245 ± 0.67 a pH 6.65 ± 0.22 a 6.66 ± 0.19 a 6.63 ± 0.18 a 6.61 ± 0.09 a Color L * 83.31 ± 0.42 c 83.16 ± 0.27 c 86.00 ± 0.13 a 85.65 ± 0.67 b a -1.65 ± 0.83 a -1.49 ± 0.32 a -2.1 ± 0.41 a -1.76 ± 0.34 a b 5.03 ± 0.21 a 5.05 ± 0.32 a 5.45 ± 0.56 a 4.81 ± 0.42 a Notes: Different superscripts on the same line indicate significant differences (P < 0.05) Results and discussion 3.1 Physicochemical properties of goat milk The physicochemical composition of raw milk is significantly influenced by the breed of the goat. Several noteworthy differences have been reported among four different breeds available in Gorontalo: kacang, local goat, ettawa crossbreed, and Saanen crossbreed (Tables 1 and 2 ). The moisture content of goat milk showed minimal variations across different breeds, ranging from 85.32–89.01% (Table 2 ). This is noteworthy since moisture is a key factor in regulating the cost, microbiological stability, and quality of milk products (p < 0.05).. The solution and colloidal suspension of additional milk ingredients are mediated by water. [ 27 ]. The findings of this study align with those of Mohsin et al. (2019), who reported moisture contents in goat milk from British Alpine, Jamnapari, Saanen, Shami, and Toggenburg breeds as 87.29%, 85.32%, 88.08%, 88.33%, and 89.01% respectively. The Jamnapari breed exhibited a lower moisture content of 85.32% compared to other breeds such as Shami, Saanen, and Toggenburg [ 28 ]. Water acts as a mediator in the solution and colloidal suspension of other milk constituents [ 29 ], [ 30 ]. Lactose content did not exhibit significant variation among the breeds ( p > 0.05), ranging from 4.26–4.55%. This uniformity suggests that lactose synthesis is largely conserved across breeds, consistent with findings by Park et al. (2007), who reported that genetic differences have a minimal effect on lactose concentration in goat milk [ 31 ]. Lactose plays a critical role in milk osmoregulation, influencing the overall solute balance and milk yield [ 32 ]. Furthermore, lactose levels in the four goat breeds studied were higher than the 3.69–3.74% reported by Setiawan et al. [ 33 ]. Lactose is the primary carbohydrate in milk, with goat milk containing 0.2–0.5% less lactose than cow milk [ 33 ]. Several factors influence lactose levels, including the nutrient composition of the feed given to livestock. Poor feed quality can result in lower lactose concentrations in milk[ 34 ] [ 35 ]. In fermented milk, lactose provides lactic acid bacteria (LAB) with energy and is essential to generate acid during fermentation. [ 36 ]. The four goat breeds' milk had fat contents ranging from 4.08–4.12%. According to the findings, the milk fat content of the four goat breeds was lower for Etawa goats (5.98–6.98%) and Jamnapari/Etawa goats (4.61–5.17%) [ 36 ] [ 37 ]. Fat content is influenced by several factors such as feeding types such as forage and concentrates. Providing forage will affect the formation of fat because forage is a source of fiber. The production of acetate significantly influences the synthesis of fatty acids, which in turn impacts the fat content in milk [ 38 ]. When livestock consume forage, it undergoes a fermentative process in the rumen by rumen microbes, resulting in the formation of Volatile Fatty Acids (VFA) such as propionate, acetate, and butyrate. Acetate is then absorbed into the bloodstream and converted into fatty acids, which are subsequently transported into the secretory cells of the udder, contributing to the production of milk fat [ 39 ]. It's important to note that both fat content and protein are key factors in determining the selling price of milk [ 40 ]. Interestingly, all four goat milk products have consistent fat content ( p > 0.05), underscoring the uniformity in this aspect of milk production. Protein content, however, exhibited more pronounced variations ( p < 0.05)., with Kacang and Local Goat breeds showing lower values (3.75% and 3.95%, respectively) compared to Etawa Crossbreed (4.11%) and Saanen Crossbreed (4.23%). The higher protein content in crossbreeds may be attributed to selective breeding practices aimed at improving milk yield and quality [ 41 ]. Protein concentration is a crucial determinant of milk processing characteristics, particularly in cheese production, where higher protein content enhances curd formation and yield [ 42 ]. The protein content in milk is directly influenced by the protein content in the feed, with higher dietary protein levels resulting in increased milk protein secretion. The primary source of protein in animal feed typically comes from concentrates [ 43 ]. Enhancing the availability of amino acids in feed has been shown to promote milk protein synthesis [ 44 ]. The production of milk protein starts when animals eat concentrate-based diet, which is then converted into amino acids by rumen bacteria. These amino acids are then absorbed in the small intestine, transported into the bloodstream, and eventually delivered to the secretory cells of the udder, where they contribute to milk protein synthesis [ 44 ]. Specific gravity serves as a key indicator of milk composition, reflecting the concentration of dissolved solids such as proteins, fats, and lactose. The recorded values ranged from 1.03320 to 1.03540, with the highest specific gravity observed in Local Goat milk (1.03540 ± 0.09) and the lowest in Kacang goat milk (1.03320 ± 0.11). However, statistical analysis indicated that these differences were not significant ( p > 0.05), suggesting that all breeds exhibit relatively similar concentrations of milk solids. The specific gravity of milk from kacang goat is 1.030 [ 11 ], besides the specific gravity of milk from etawa crossbreed 1.030 and saanen crossbreed 1.0295 [ 45 ]. This could be influenced by the fat content of milk which has a negative impact on the specific gravity of milk [ 46 ]. According to McCarthy and Singh [ 47 ] stated that the specific gravity of milk depends on the fat content and solid material of the milk, because the specific gravity of fat is lower than the specific gravity of density of water or milk plasma. The release of gases such as carbon dioxide (CO 2 ) and nitrogen (N 2 ) into milk immediately following the milking process can lead to an increase in specific gravity. Understanding this phenomenon is essential for monitoring milk quality and ensuring optimal processing conditions [ 48 ]. Goat milk has a specific gravity that is lower than sheep milk (range: 1.0347–1.0384 kg/m 3 ) but higher than cow's milk (range: 1.0231–1.0398 kg/m 3 ) [ 31 ]. There was no discernible difference in the pH of the milk from the four goat breeds (p > 0.05). Milk from various goat breeds has a pH value that ranges from 6.61 to 6.65, which suggests possible milk damage [ 49 ]. The precise pH value is influenced by components present in fresh milk, such as CO 2 , phosphate, citrate, and protein, which affect the milk's ability to resist changes in pH and acidity, thus preventing milk spoilage [ 50 ]. Bacterial activity can cause the pH to become more acidic, dropping below the average value of 6.5–6.7 [ 51 ]. pH values higher than 6.7 usually indicate the possibility of mastitis [ 52 ]. The colour characteristics of goat milk from four genotypes—Kacang, Local Goat, Etawa Crossbreed, and Saanen Crossbreed—were evaluated using the CIE L a b* system, where L* denotes lightness, a* indicates the green–red axis, and b* represents the blue–yellow axis. Among the genotypes, Etawa Crossbreed milk exhibited the highest L* value (86.00 ± 0.13), indicating a significantly lighter appearance compared to the other groups (p < 0.05), followed by Saanen Crossbreed (85.65 ± 0.67). These findings suggest a superior visual brightness, which is often associated with higher consumer appeal and milk freshness [ 53 ]. In contrast, Kacang and Local Goat milk showed lower lightness values (both 83.2–83.3), indicating a comparatively darker hue. The higher L* values in goat milk are related to smaller fat globule size and more complete conversion of β-carotene into colorless vitamin A, resulting in a whiter appearance [ 54 ]. This whitening effect enhances light scattering, making crossbred goat milk visually more appealing to consumers. The a* values across all genotypes were negative, ranging from − 1.49 to − 2.10, which indicates a slight shift toward the green spectrum. While these differences were not statistically significant (p > 0.05), they align with the findings of Milovanovic et al [ 53 ], who reported higher greenish tones in goat and sheep milk compared to cow milk. The greenish hue is attributed to the lack of residual carotenoids, a trait unique to goat species due to their metabolic conversion pathway [ 55 ][ 54 ]. Regarding b* values, all breeds showed mild yellowish tones (b* = 4.81–5.45), with Etawa Crossbreed having the highest yellow component. The yellow appearance may be attributed to residual carotenoids or fat-soluble pigments, although values were lower than typically seen in cow milk, consistent with literature noting the lack of β-carotene conversion in goat milk [ 56 ]. Overall, the colour traits observed reinforce the role of genotype in influencing milk visual attributes, with crossbred goats (especially Etawa) producing milk that appears brighter and slightly more yellow—features that may enhance its marketability and consumer preference, as confirmed by both instrumental and visual assessments in Milovanovic et al [ 53 ]. 3.2 Amino acid and protein profile The protein composition of goat milk from several breeds in Gorontalo, including Kacang, Local Goat, Etawa Crossbreed, and Saanen Crossbreed, was displayed in the SDS-PAGE (Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis) image (Fig. 1). The molecular weight distribution and relative abundance of casein and whey protein fractions, which are intimately related to the goat milk's amino acid composition, can be learned from the SDS-PAGE analysis of goat milk proteins (Table 1 ). Breed-specific differences in protein expression are confirmed by the protein bands seen in the SDS-PAGE gel, especially in β-casein, α-casein, and κ-casein (25–35 kDa), as well as β-lactoglobulin and β-lactalbumin (14–18.4 kDa). These variations are consistent with the amino acid profiles found in the milk of the Saanen, Etawa, Kacang, and Local goat breeds.. The primary protein component of goat milk, caseins are necessary for both the stability of the milk and the creation of cheese [ 57 ]. The Fig. 1 results show that Etawa and Saanen Crossbreeds exhibit more intense casein bands, indicating higher casein concentrations compared to Kacang and Local Goat breeds. This observation correlates with the higher glutamic acid (Glu) and aspartic acid (Asp) concentrations in these breeds, as shown in the amino acid composition (Table 1 ). Aspartic acid (Etawa: 3.14 g/100 g, Saanen: 2.15 g/100 g) along with glutamic acid (Etawa: 7.02 g/100 g, Saanen: 7.14 g/100 g) are known for their role in stabilizing protein structures and enhancing the emulsification properties of milk [ 57 ]. Additionally, higher levels of proline (Pro) and threonine (Thr) in Etawa and Saanen Crossbreeds further support their superior casein content [ 58 ]. Proline (Etawa: 5.02 g/100 g, Saanen: 3.11 g/100 g) enhances protein stability, while threonine (Etawa: 6.24 g/100 g, Saanen: 4.11 g/100 g) is essential for protein synthesis and milk viscosity, aligning with the observed rheological differences among the breeds. Whey proteins, particularly β-lactoglobulin and β-lactalbumin, play a significant role in immune function, digestibility, and processing properties [ 59 ]. Etawa and Saanen Crossbreeds have stronger whey protein bands, according to Fig. 1 data, which is in keeping with their greater levels of branched-chain amino acids (BCAAs), including valine (Val), isoleucine (Ile), and leucine (Leu). These amino acids are critical for protein metabolism and muscle development, making the milk from these breeds more suitable for high-protein formulations and sports nutrition products [ 60 ]. Specifically, Etawa Crossbreed showed the highest levels of leucine (2.44 g/100 g), isoleucine (2.12 g/100 g), and valine (3.14 g/100 g), aligning with the observed abundance of β-lactoglobulin in its SDS-PAGE profile. Saanen Crossbreed also demonstrated high levels of these amino acids, reinforcing its potential for nutritional applications, particularly in infant formula and dietary supplements [ 61 ]. 3.3 Extended protein fraction estimation in goat milk from four genotypes Protein composition is one of the most critical determinants of milk quality, influencing its nutritional value, processing behavior, and functional applications. The main protein fractions—β-casein, α-casein, κ-casein, β-lactoglobulin, and β-lactoalbumin—were estimated in this work using SDS-PAGE electrophoresis (Fig. 1) and densitometric analysis in goat milk obtained from four genotypes: Kacang, Local Goat, Etawa Cross, and Saanen Cross. These estimations were calculated using densitometric band intensity data ((in arbitrary units, A.U.)) and translated into approximate concentrations (g/L), Table 3 following a conversion approach similar to that of Li et al [ 23 ], which assumes a linear relationship between band intensity and protein concentration under Coomassie Brilliant Blue staining. Table 3 Protein Fraction of Goat Milk Genotype β-Casein (g/ L) α-Casein (g/ L) κ-Casein (g/ L) β-Lactoglobulin (g/ L) β-Lactalbumin (g/ L) Kacang 2.82 ± 0.31 c 0.98 ± 0.17 c 0.72 ± 0.43 b 4.34 ± 0.56 a 1.29 ± 0.11 a Local Goat 4.14 ± 0.22 b 1.43 ± 0.45 b 1.02 ± 0.21 a 3.93 ± 0.11 a 1.17 ± 0.11 a Etawa Crossbreed 5.16 ± 0.08 a 1.78 ± 0.34 a 1.27 ± 0.18 a 3.65 ± 0.11 a 1.08 ± 0.11 a Saanen Crossbreed 5.62 ± 0.76 a 1.96 ± 0.09 a 1.43 ± 0.91 a 3.45 ± 0.11 a 1.03 ± 0.11 a Notes: Different superscripts on the same column indicate significant differences (P < 0.05) Among the genotypes evaluated, Saanen Cross demonstrated the highest total protein concentration (9.05 g/L), primarily due to elevated casein levels. This group showed 5.6 g/L of β-casein, alongside estimated concentrations of 1.96 g/L of α-casein and 1.4 g/L of κ-casein, making it the most casein-rich milk type in this study. The dominance of β-casein, comprising over 60% of the total protein, aligns with findings from Costa et al [ 32 ], who also reported Saanen milk to contain high levels of β- and αs1-caseins. These fractions are essential for curd formation, texture development, and yield in cheese-making processes [ 62 ]. Conversely, Kacang goat milk presented a markedly different profile, with a total protein content of 7.1 g/L, of which β-lactoglobulin accounted for 4.3 g/L—the highest among all breeds. Its estimated β-casein concentration was 2.8 g/L, with accompanying levels of 0.98 g/L α-casein and 0.7 g/L κ-casein. This composition reflects a whey-dominant profile, suitable for applications in specialized nutrition, including infant formula or medical nutrition products due to higher digestibility and lower allergenicity [ 63 ]. The relatively high β-lactoalbumin content (1.29 g/L) further enhances this nutritional suitability, given its known role in lactose synthesis and immune functions. Etawa Cross and Local Goat displayed intermediate profiles. Etawa Cross milk contained 5.1 g/L β-casein, with corresponding values of 1.79 g/L α-casein and 1.28 g/L κ-casein. Local Goat milk showed 4.1 g/L β-casein, 1.44 g/L α-casein, and 1.03 g/L κ-casein. Both genotypes had balanced whey and casein fractions, making them versatile for both processing and dietary formulations [ 64 ]. Notably, their protein-to-casein ratios and viscosity values positioned them between the Kacang and Saanen genotypes, suggesting multifaceted industrial potential. Overall, the extended fractionation of goat milk proteins provides a nuanced understanding of compositional diversity among breeds. The casein-to-whey ratio, along with individual casein subtype estimates, directly informs processing decisions. Higher casein levels favor curd firmness and yield, while elevated whey protein content enhances biofunctionality. These insights underscore the importance of genotype selection in dairy breeding programs and the design of targeted dairy products for specific nutritional and technological purposes. 3.4 Zeta potential and particle size Zeta potential is an important measure for understanding the stability and charge of milk systems. This study looked at the zeta potential values in goat milk from four types of goats: Kacang, Local Goat, Etawa Crossbreed, and Saanen Crossbreed. The values ranged from − 8.48 to − 8.65 mV (Table 2 ). The Etawa Crossbreed had the most negative value at − 8.65 ± 0.33 mV, suggesting better stability due to stronger electrostatic repulsion. The lowest negative value was − 8.48 ± 0.22 mV for Kacang goat milk, although the differences were not statistically significant (p > 0.05). These findings match earlier research by Li et al [ 24 ], which found raw goat milk zeta potential values between − 8.50 and − 8.75 mV. However, our results differ from Huo et al [ 65 ], who reported a value of − 27.5 mV for Aote Laoshan goat milk, and Chen et al [ 66 ], who found a value of − 30.3 mV for raw goat milk. The difference in zeta potential may be because of variations in milk composition related to different goat breeds, especially in proteins and fats. The charge of the micellar particles in milk is influenced by the casein and whey proteins. Glutamic and aspartic acids are among the many negatively charged amino acids found in caseins, especially β-casein and αs1-casein. Past research indicates that crossbred goats, such as Saanen and Etawa, usually produce milk with more casein and richer in these acids, which may lead to more negative zeta potential values [ 20 ]. Additionally, differences in the milk fat globule membrane (MFGM) and the size of fat globules may also affect the surface charge. Goats that are crossbred typically produce milk with smaller fat globules, increasing the milk's surface area.. This exposes more charged phospholipids and glycoproteins on the MFGM, increasing electrostatic repulsion. In contrast, goats may have larger fat globules with different membrane compositions, resulting in a reduced surface charge and a less negative zeta potential [ 67 ]. Mineral content, especially calcium and phosphate ions, can also affect the surface charge because they bind to casein micelles. While generally minor, differences in ionic composition among goat breeds can lead to small variations in their behavior in an electric field [ 68 ]. Goat milk samples' particle size distribution was also assessed in order to identify structural traits that are important for stability and digestion. Table 2 explain the particle diameters ranged narrowly between 244 and 247 nm across the four genotypes, with no significant statistical differences (p > 0.05), indicating a relatively consistent colloidal system regardless of genotype. The size range and distribution properties of the particles in the protein solution are reflected in the particle size distribution. Raw goat milk protein has a wide and irregular bimodal particle size distribution, with most of the protein particles localized around 200 nm, as demonstrated by Huo et al. [ 66 ]. Comparative data from Li et al [ 24 ] support this, showing that raw goat milk averaged 250 nm. Because different breeds differ in their milk composition, especially in terms of fat and protein content, as well as their molecular structures, which are impacted by breed-specific genetics, food, and lactation physiology, the particle size of raw goat milk can also vary [ 67 ]. Protein interaction and fat globule size: Goat milk is easier to digest than cow milk because it naturally contains fewer fat globules and less αS1-casein. [ 68 ]. However, the size and uniformity of fat globules and protein aggregates still vary by breed due to inherent genetic and biochemical differences. For example, differences in casein micelle structure and whey protein concentration between breeds affect aggregation and emulsification behavior, leading to different particle size distributions​. Breed influence on milk composition; goat milk composition—including the types and amounts of caseins and whey proteins—differs between breeds. These compositional differences affect thermal stability and response to processing, ultimately influencing particle size and emulsion stability​. The particle size distribution (e.g., bimodal vs. uniform) and changes in zeta potential indicate breed-dependent responses in milk microstructure when subjected to processing. Ultrasonic treatment, for instance, results in more uniform and smaller particles, but the starting size and distribution in raw milk still reflect breed-specific characteristics​. Thus, differences in particle size in raw goat milk from various breeds arise primarily due to variations in fat globule size, protein content and structure, and overall milk composition, all of which are inherently linked to the genetic traits of each goat breed. 3.5 Rheological behavior of goat milk Shear rate is a fundamental parameter in rheological analysis that describes the deformation rate of a fluid under applied stress. In dairy science, understanding the shear rate behavior of goat’s milk is critical for optimizing its processing, stability, and texture in various dairy applications. Goat’s milk, like other dairy fluids, can exhibit non-Newtonian flow behavior, depending on its composition. Figure 2 shows a non-linear, goat’s milk exhibits non- Because of the existence of fat globules, casein micelles, and protein structures, Newtonian features are more prevalent [ 69 ] [ 70 ]. The viscosity decreases as shear rate increases, which is a key property of goat’s milk due to its colloidal nature. At high shear rates; a declining viscosity trend with increasing shear rate would confirm pseudoplastic behavior, meaning the milk becomes thinner as shear increases. This is a desirable property in dairy processing, making milk easier to pump, mix, and homogenize without phase separation. The shear rate response of goat’s milk is influenced by breed differences: Etawa and Saanen Crossbreeds typically show higher shear resistance due to increased protein and total solids content, resulting in a more structured milk network. Kacang and Local Goat breeds tend to have lower viscosity and faster shear response, indicating a more fluid-like consistency with smaller fat globules and lower solid content. Implications for dairy processing and stability; understanding shear rate behavior ensures optimal homogenization conditions, preventing fat separation in processed milk. Shear-thinning properties are beneficial for products like yogurt and cheese, where controlled viscosity is required for desirable texture and mouthfeel. Milk with lower shear resistance may experience phase separation faster, requiring stabilization techniques for extended shelf life. Viscosity is a crucial rheological parameter that determines the flow behavior and textural properties of goat’s milk, influencing its processing, stability, and sensory perception. The analysis of viscosity in Fig. 3 provides insights into the physical characteristics of goat’s milk in Gorontalo, which are affected by breed composition, fat and protein content, and processing conditions. In general, goat's milk has non-Newtonian fluid behavior, meaning that as the shear rate increases, its viscosity falls, suggesting pseudoplastic (shear-thinning) qualities [ 70 ]. This is characteristic of milk due to the presence of casein micelles and fat globules that align under shear stress, reducing resistance to flow. The data in Fig. 3 show a declining viscosity trend with increasing shear rate, it confirms the pseudoplastic nature of goat’s milk, similar to previous studies on dairy rheology. Because goat breeds range in terms of fat globule size and protein concentration, the viscosity of their milk can vary greatly, and total solids content. Typically; higher viscosity is associated with higher protein and fat content, as seen in breeds such as Etawa and Saanen Crossbreeds. Lower viscosity is observed in milk with lower total solids and smaller fat globules, which is characteristic of Kacang and Local Goat breeds. Figure 3 demonstrates variations among breeds, it suggests that compositional differences play a major role in determining viscosity. Milk viscosity is highly temperature-dependent, with lower temperatures increasing viscosity due to reduced molecular mobility [ 66 ]. It is expected that higher temperatures result in decreased viscosity, following the Arrhenius equation for fluid dynamics [ 71 ]. This is essential for optimizing processing conditions such as pasteurization, homogenization, and ultrafiltration in dairy industries. 3.6 Microstructure analysis The microstructural features of goat milk from several breeds in Gorontalo, including Kacang, Local Goat, Etawa Crossbreed, and Saanen Crossbreed, are depicted in the fluorescence microscope photos above (Fig. 4).This analysis provides insights into the fat globule distribution, protein network, and overall structural differences, which directly influence milk stability, digestibility, and dairy processing potential. The size and distribution of fat globules play a critical role in determining milk texture, mouthfeel, and processing behavior [ 72 ]. Kacang and Local Goat milk show a denser distribution of smaller fat globules, suggesting better emulsion stability and easier digestibility, which aligns with findings that smaller globules improve lipid absorption [ 73 ]. Etawa and Saanen Crossbreed milk exhibit larger and more heterogeneous fat globules, indicating a higher fat content. Implications for dairy processing; higher fat globule uniformity in Kacang and Local Goat milk makes it better suited for fluid milk consumption and products requiring emulsion stability, such as yogurt and infant formula. Larger fat globules in Etawa and Saanen Crossbreed milk contribute to richer texture and creamier consistency, making them ideal for cheese production, butter, and high-fat dairy products. The differences in fat globule structure among breeds highlight the need for specific processing techniques, such as homogenization, pasteurization, and fat standardization, to optimize dairy product quality. 3.7 Principal component analysis (PCA) The PCA biplot provides a comprehensive visual representation of how different breeds of goats are distributed based on their physical milk composition. It illustrates the relationships among the measured variables and how they contribute to the overall variation in physical milkFigure 5A displays the plot of the PCA model of physical milk composition, while Fig. 5 B displays the plot of the PCA model of chemical milk composition. Eighty-two percent of the variations in the data set were displayed by the first two PCs (PC1 and PC2) (Fig. 5 A). The PC1 was most affected by the density (specific gravity) and brightness (L) of the milk, accounting for 52.94%.. The PC2 accounted for 29.06% of the captures variations in Zeta Potential, pH, and minor color properties. The analysis of the samples in quadrant I (PC1+, PC2+) showcases the outstanding qualities of milk from the Etawa crossbreed. With the highest L values indicating brightness and a larger particle size, this breed emerges as one of the most favorable options for dairy processing, underscoring its potential to make a significant contribution to the industry. In quadrant II (PC1-, PC2+), we find valuable insights from the milk of the Kacang goat. This breed plays a notable role in the overall variation in milk properties, characterized by a higher specific gravity and lower lightness (L), along with a distinctive Zeta Potential that reflects the electrical charge on its milk particles. These characteristics suggest that Kacang goat milk may be particularly well-suited for concentrated dairy products, highlighting the breed's importance in specialized dairy applications. Quadrant III (PC1-, PC2-) includes milk from the Local goat, which exhibits lower L* values and potentially different specific gravity or a* values. This indicates that the milk from the Local goat is versatile and well-suited for general dairy applications, including pasteurized and fresh milk, thereby enabling a broader range of dairy products. Finally, the samples in quadrant IV (PC1+, PC2-) present milk from the Saanen breed. While it shares similarities with Etawa milk, it displays slightly lower brightness and a moderate particle size. Saanen milk offers a robust option for standard dairy applications that demand high-quality and stable products. By understanding these unique properties, producers can make informed decisions that effectively enhance their dairy offerings. Plotting of the PCA model of chemical milk composition is displayed in Fig. 5 B.. 92.71% of the variations in the data set were displayed by the first two PCs (PC1 and PC2). Moisture content, lactose, fat, and protein composition were the main factors influencing the PC1, which explained 78.73% of the overall variance.. With the exception of moisture content, which was found at PC1's negative loading, all of these variables showed positive correlations. The amounts of amino acids (Asp, Glu, Thr, Val, etc.) were the main cause of the PC2's 13.98% variance. Known for its outstanding quality, milk from the Etawa crossbreed is highlighted in Quadrant I (PC1+, PC2+).. This quadrant is defined by high moisture content, elevated protein levels, and a rich array of essential amino acids, including glutamic acid (Glu), aspartic acid (Asp), and branched-chain amino acids like leucine, isoleucine, and valine. The strong correlation between increased protein and these amino acids indicates that Etawa milk is highly nutritious, making it suitable for functional dairy products such as fortified yogurts and premium cheeses. Quadrant II (PC1-, PC2+) presents Kacang goat milk, characterized by lower moisture and protein content but higher fat concentration and amino acids like cysteine (Cys) and methionine (Met). The inverse relationship between these amino acids and moisture suggests that Kacang milk has a higher dry matter concentration, making it particularly advantageous for rich cheeses and butter. Quadrant III (PC1-, PC2-) features milk from local goats, which, while lower in protein and amino acids, has moderate moisture and lactose levels. It remains valuable for fresh goat milk products and traditional fermented items, benefiting from a mild flavor. Quadrant IV (PC1+, PC2-) showcases Saanen crossbreed milk, which shares beneficial traits with the Etawa breed. Though it has slightly lower concentrations of select amino acids, it retains high moisture and protein content, making it suitable for high-nutritional-value dairy products, including premium cheeses and quality cream. Dairy producers may strategically maximize milk utilization, improve product quality, and satisfy changing market demands by acknowledging the distinctive qualities of each breed. To improve our understanding of milk quality and its market potential, future research should focus on several key areas. First, investigating the impact of dietary composition on the physicochemical properties of milk could optimize nutrient intake and enhance product stability. Additionally, assessing the influence of various lactation stages on milk composition through Principal Component Analysis (PCA) would provide valuable insights into the dynamic changes in milk quality over time. Furthermore, exploring consumer preferences regarding goat milk characteristics could help align dairy product development with market demand. Future studies should also examine the combined effects of diet and lactation stage on the chemical composition of milk. Adapting processing methods to accommodate breed-specific variations in milk properties may further improve both its nutritional value and commercial viability. Moreover, establishing market segmentation strategies based on milk composition could optimize product quality and maximize economic benefits in the dairy industry. Conclusion This study presents a comprehensive characterization of the physicochemical and rheological properties of raw goat milk from four genotypes—Kacang, Local Goat, Etawa Crossbreed, and Saanen Crossbreed—reared in Indonesia. The results demonstrated that genotype significantly influences milk composition, particularly in protein content, amino acid profiles, micellar protein structure, and rheological behavior. Crossbreeds, notably Saanen and Etawa, produced milk with higher total protein and casein fractions, contributing to enhanced viscosity and suitability for cheese and fermented dairy production. Conversely, Kacang goat milk exhibited higher whey protein content and sulfur-rich amino acids, aligning with applications in specialized nutrition. Despite breed-dependent differences in microstructure, all milk types displayed consistent pseudoplastic behavior and colloidal stability, as confirmed by zeta potential and particle size measurements. These findings highlight the functional diversity of goat milk in Indonesia and support targeted valorization strategies for each breed. This study enriches the existing knowledge base by providing molecular and rheological insights that can guide breed selection, dairy processing optimization, and functional food development. Future research should investigate the effects of feed and lactation stage on milk quality and explore consumer preferences to bridge scientific potential with market viability. Declarations Funding Funding by the Research Institute for Humanity and Nature (RIHN) Japan, No 14200102. Data availability Data is provided within the manuscript or supplementary information files Clinical trial registration Not applicable Ethics approval and consent to participate Permission to conduct this research was obtained from the Faculty Board of the Faculty of Agriculture, State University of Gorontalo, in accordance with the standard operating procedures established by the institution’s Research and Development Committee. This study did not involve the use or handling of live animals, including breeding or maintenance within an animal facility. The procedures for the collection, processing, and utilization of milk materials in this study adhered to all applicable local, national, and international ethical guidelines for the use of animal-derived products in scientific research. Consent for publication Not applicable. Competing interests The authors declare no competing interests. References J. 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Vishveshwara, ‘Effect of constraints by threonine on proline containing αa-helix—A molecular dynamics approach’, Biophys Chem , vol. 46, no. 1, pp. 77–89, Feb. 1993, doi: 10.1016/0301-4622(93)87009-L. R. Sharma, ‘Whey Proteins in Functional Foods’, Whey Proteins: From Milk to Medicine , pp. 637–663, Jan. 2019, doi: 10.1016/B978-0-12-812124-5.00018-7. X. Dong et al. , ‘Increasing the availability of threonine, isoleucine, valine, and leucine relative to lysine while maintaining an ideal ratio of lysine:methionine alters mammary cellular metabolites, mammalian target of rapamycin signaling, and gene transcription’, J Dairy Sci , vol. 101, no. 6, pp. 5502–5514, Jun. 2018, doi: 10.3168/jds.2017-13707. H. Kumar, D. Yadav, N. Kumar, R. Seth, and A. Goyal, ‘Nutritional and nutraceutical properties of goat milk - A review’, Indian Journal of Dairy Science , vol. Accepted, Apr. 2016. D. A. Goulding, P. F. Fox, and J. A. O’Mahony, ‘Milk proteins: An overview’, Milk Proteins: From Expression to Food , pp. 21–98, Nov. 2019, doi: 10.1016/B978-0-12-815251-5.00002-5. K. K. S. Borba et al. , ‘Characterization and biological activity of ultrafiltrate goat whey protein concentrate over the in vitro digestion’, Food Biosci , vol. 65, p. 106087, Mar. 2025, doi: 10.1016/J.FBIO.2025.106087. U. Sadiq, H. Gill, and J. Chandrapala, ‘Casein micelles as an emerging delivery system for bioactive food components’, Foods , vol. 10, no. 8, Aug. 2021, doi: 10.3390/FOODS10081965. W. Hou et al. , ‘Impact of ultrasonic and heat treatments on the physicochemical properties and rennet-induced coagulation characteristics of milk from various species’, Ultrason Sonochem , vol. 111, Dec. 2024, doi: 10.1016/j.ultsonch.2024.107084. X. Chen et al. , ‘A Comparative Study of Ultrasound and Thermal Processing: Effects on Stability and Protein Structure in Goat Milk’, J Dairy Sci , Jan. 2025, doi: 10.3168/jds.2024-25918. G. Liao et al. , ‘Comparison of the Lipid Composition of Milk Fat Globules in Goat (Capra hircus) Milk during Different Lactations and Human Milk’, Foods , vol. 13, no. 11, Jun. 2024, doi: 10.3390/foods13111618. M. Corredig, P. K. Nair, Y. Li, H. Eshpari, and Z. Zhao, ‘Invited review: Understanding the behavior of caseins in milk concentrates’, J Dairy Sci , vol. 102, no. 6, pp. 4772–4782, Jun. 2019, doi: 10.3168/JDS.2018-15943. H. A. Pushpadass, F. M. E. Emerald, B. V Balasubramanyam, and S. S. Patel, ‘11 - Rheological Properties of Milk-Based Beverages’, in Milk-Based Beverages , A. M. Grumezescu and A. M. Holban, Eds., Woodhead Publishing, 2019, pp. 373–396. doi: https://doi.org/10.1016/B978-0-12-815504-2.00011-6. H. F. George and F. Qureshi, ‘Newton’s Law of Viscosity, Newtonian and Non-Newtonian Fluids’, in Encyclopedia of Tribology , Q. J. Wang and Y.-W. Chung, Eds., Boston, MA: Springer US, 2013, pp. 2416–2420. doi: 10.1007/978-0-387-92897-5_143. A. Messaâdi et al. , ‘A New Equation Relating the Viscosity Arrhenius Temperature and the Activation Energy for Some Newtonian Classical Solvents’, E-Journal of Chemistry , vol. 2015, pp. 1–15, May 2015, doi: 10.1155/2015/163262. N. Tzirkel-Hancock, L. Sharabi, and N. Argov-Argaman, ‘Milk fat globule size: Unraveling the intricate relationship between metabolism, homeostasis, and stress signaling’, Biochimie , vol. 215, pp. 4–11, Dec. 2023, doi: 10.1016/J.BIOCHI.2023.10.003. Y. Wang et al. , ‘Size-dependent composition and in-situ structure analysis of the milk fat globule membrane in buffalo milk’, Food Chem , vol. 464, p. 141766, Feb. 2025, doi: 10.1016/J.FOODCHEM.2024.141766. Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6429174","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":467186709,"identity":"1b254b3b-18bf-4e33-9ef4-5dbcdf8de499","order_by":0,"name":"Agus Bahar Rachman","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA60lEQVRIiWNgGAWjYBCDBCBmfADjHeAhUguzAcla2CTgXHxa+PvXGH+uqLHL020/e6yad0ctA3/7AcYDb/BokbjxxkzyzLHkYrMzeWm3ec8cZ5A4k8BwcA4+F904Y8bYwHYgcduBHLPbvG3HgCIMDIfxOUz+xhnjjw3/gFrOvzErBmmRJ6TF4HyPgWRjG1DLjRwzZt62GgYDQloMb7CVSTb2JQO1vDGWnNt2gMfwTGIDXr/InT+8+WPDNzugw3IMP7xtq5OTO3748Ad8IcYgkYDCBTmJsQGfBmDEHEDh1uFXPQpGwSgYBSMSAAALUFYjCwErYAAAAABJRU5ErkJggg==","orcid":"","institution":"State University of Gorontalo","correspondingAuthor":true,"prefix":"","firstName":"Agus","middleName":"Bahar","lastName":"Rachman","suffix":""},{"id":467186710,"identity":"64e3badc-0eae-456a-a712-8004574a7043","order_by":1,"name":"Fahrul Ilham","email":"","orcid":"","institution":"State University of Gorontalo","correspondingAuthor":false,"prefix":"","firstName":"Fahrul","middleName":"","lastName":"Ilham","suffix":""},{"id":467186711,"identity":"4d2f0dd1-eff3-40ef-a3fe-950a7ef81820","order_by":2,"name":"Lukman Hakim","email":"","orcid":"","institution":"National Research and Innovation Agency","correspondingAuthor":false,"prefix":"","firstName":"Lukman","middleName":"","lastName":"Hakim","suffix":""},{"id":467186712,"identity":"11e9b9e0-aab3-4857-b6b2-a75c1b2c9965","order_by":3,"name":"Nicolays Jambang","email":"","orcid":"","institution":"National Research and Innovation Agency","correspondingAuthor":false,"prefix":"","firstName":"Nicolays","middleName":"","lastName":"Jambang","suffix":""},{"id":467186713,"identity":"ef10c45a-150b-45d9-82e4-9d9ed0908d91","order_by":4,"name":"Andi Patiware Metaragakusuma","email":"","orcid":"","institution":"Research Institute for Humanity and Nature","correspondingAuthor":false,"prefix":"","firstName":"Andi","middleName":"Patiware","lastName":"Metaragakusuma","suffix":""}],"badges":[],"createdAt":"2025-04-11 14:23:26","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6429174/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6429174/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":84206550,"identity":"af287eec-df0a-4c83-a1aa-755060011098","added_by":"auto","created_at":"2025-06-09 09:12:29","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":123544,"visible":true,"origin":"","legend":"\u003cp\u003eLegend not included with this version\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6429174/v1/d36d6b627a35d51e19ddf975.png"},{"id":84206552,"identity":"5857b0e1-35e9-43b8-b8d9-5ab5a235291d","added_by":"auto","created_at":"2025-06-09 09:12:29","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":104764,"visible":true,"origin":"","legend":"\u003cp\u003eThe shear rate of goat milk\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6429174/v1/7f538c6ca41bbcfe7a8c6dfe.png"},{"id":84206554,"identity":"bc798aea-9007-43aa-8980-8a75b03503dd","added_by":"auto","created_at":"2025-06-09 09:12:29","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":64243,"visible":true,"origin":"","legend":"\u003cp\u003eThe viscosity of goat milk\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6429174/v1/e0398cf09c6eb1f7e6394586.png"},{"id":84206559,"identity":"a4eaf562-265a-4c32-a3eb-4e2bf3b95c04","added_by":"auto","created_at":"2025-06-09 09:12:29","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":1158580,"visible":true,"origin":"","legend":"\u003cp\u003eLegend not included with this version\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6429174/v1/c6dc86c1c6bf01c01aa0800e.png"},{"id":84207709,"identity":"61b132df-fbb6-4800-a5d5-1a185667fbbc","added_by":"auto","created_at":"2025-06-09 09:28:29","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":819856,"visible":true,"origin":"","legend":"\u003cp\u003ePrincipal Component Analysis (PCA); a) physical properties, and b) chemical properties\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-6429174/v1/05365154c8ed4f6c490fd145.png"},{"id":84208041,"identity":"c494b299-44ed-4e0c-83e6-f7b9b30a455f","added_by":"auto","created_at":"2025-06-09 09:36:31","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3429511,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6429174/v1/fe00a569-bb17-48bb-a058-6ec4613c659a.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Uncovering the Functional Potential of Goat Milk: Physicochemical and Rheological Comparison Across Local Goats in Gorontalo Indonesia","fulltext":[{"header":"Introduction","content":"\u003cp\u003eDairy goat farming in Indonesia has emerged as a promising sector, evidenced by a notable increase in the goat population of 2.89% in 2021, bringing the total to 19.23\u0026nbsp;million goats [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. This upward trend reflects a growing awareness of the many benefits associated with goat milk, which is gaining recognition for its superior nutritional qualities [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Research indicates that goat milk offers a more favorable nutritional profile compared to cow milk, primarily due to its smaller fat globules. This unique characteristic not only enhances its creamy texture but also facilitates easier digestion [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Goat milk also has a higher content of conjugated linoleic acid and medium-chain fatty acids, which add to its unique flavor and certain health advantages [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Its impressive protein content, along with its buffering capacity and alkalinity, makes it particularly beneficial for individuals seeking to manage stomach ulcers [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e] In addition, goat milk's peculiar casein micelle structure is less solvated and more heat stable than cow milk's, which is very beneficial in the dairy industry, especially when manufacturing cheese. [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Overall, the growth of the dairy goat industry in Indonesia holds significant promise for both producers and consumers as the appreciation for goat milk continues to grow [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIndonesia is renowned for its diverse dairy goat breeds, each playing an integral role in providing high-quality, nutritious milk and dairy products to local communities [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Among these breeds are the indigenous Kacang and notable crossbreeds such as the Saanen Crossbreed, Etawah Crossbreed, and Saanen-Etawah (Sapera). These breeds have been carefully developed through strategic crossbreeding with varieties from South Asia, Africa, and Europe [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. The Indonesian government\u0026rsquo;s official recognition of the Etawah Crossbreed as a valuable local livestock breed highlights its commitment to preserving the nation\u0026rsquo;s agricultural heritage and fosters confidence in the future of the dairy industry [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Additionally, Saanen goat farms represent a highly effective alternative source of milk production, further enriching the local dairy sector [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. In Gorontalo, innovative crossbreeding efforts between the Etawah Crossbreed and Kacang goats have resulted in the development of Local Gorontalo goats, which have successfully adapted to their local environment [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. It is also important to emphasize the Kacang goat, an indigenous breed with significant untapped potential, which can greatly enhance dairy production in Indonesia [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. The expansion and sustainability of the dairy sector in the area will be greatly aided by identifying and fostering these capabilities. Despite the huge goat population in Gorontalo, there is a lot of room to grow the usage of goat milk as a major source of animal protein. This underutilization highlights an opportunity for growth, particularly as consumer preferences favor cow's milk, often due to its more familiar flavor compared to the distinctive taste of goat milk. Several prominent goat breeds, including Kacang, Local Goat, Saanen Crossbreed, and Etawa Crossbreed, serve a dual purpose by providing both milk and meat [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e] [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. By prioritizing high-yield and productive breeds, breeders can make strategic decisions that align with the economic goals of goat farming [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Moreover, the increasing demand for goat milk with desirable functional properties reflects a promising market opportunity. By capitalizing on this potential, the community could significantly boost the consumption of goat milk [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Establishing a Transdisciplinary Community of Practice (TDCoP) that blends scientific research, local agricultural expertise, and industry needs could facilitate this advancement [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. This collaborative approach would not only promote the sustainability of local agriculture but also enhance knowledge transformation in rural areas, benefiting both producers and consumers in Gorontalo [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThis study aims to analyze the physicochemical composition, rheology, and microstructure of raw milk from four distinct goat breeds in Indonesia. It is anticipated that there will be significant variations in the chemical and physical compositions also rheology of the milk samples due to the influence of breed, which is a crucial determinant of milk composition. These insights are essential for goat breeders and milk product manufacturers as they can enhance breed selection, cross-breeding programs, and the development of nutritionally superior functional foods. Additionally, it's important to note that other factors like feed, environmental conditions, and daily care practices may also impact the characteristics of these milk samples.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Materials\u003c/h2\u003e \u003cp\u003e Fresh goat milk samples were gathered from 48 animals representing four different breeds: Kacang, Local Gorontalo, Saanen crossbreed, and Etawa crossbreed, with 12 animals from each breed. These animals were in the 8 months of lactation stage and were at a farm in Tulabolo Village, Suwawa District, Bone Bolango Regency, Province of Gorontalo, Indonesia. Their diet consisted of sorghum stem and leaves, dairy goat pellets, and corn After being manually extracted in the morning and aseptically stored in sterile glass bottles, the milk was brought to the lab in an icebox that was kept between 2 and 4\u0026deg;C. Subsequently, the samples were preserved at \u0026minus;\u0026thinsp;20\u0026deg;C for further use.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Physicochemical Composition of Goat Milk\u003c/h2\u003e \u003cp\u003eUsing accepted practices, the samples were subjected to a comprehensive proximate composition analysis. AOAC Official Method 925 was used to compute the total solid content. 23 Milk Solids (Total) [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. The protein content were measured using the Kjeldahl method (EN ISO 8968-1:2002)[\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e], while the fat content was accurately calculated using the Gerber method (AOAC 2000.18)[\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. The specific gravity were measured using a Lactodensimeter 66110 (Funke Gerber, Germany). Furthermore, pH measurements were conducted using a standardized pH level (model Eutech\u0026trade; PC 700 Multi-parameter Meter, manufactured by Thermo Fisher Scientific, Waltham, MA, USA). with pH 4 and 7 buffer solutions.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Amino Acid Analysis by HPLC\u003c/h2\u003e \u003cp\u003eThe analysis followed the procedures outlined with some modifications[\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. The thawed milk samples A 0.2 \u0026micro;m cellulose nitrate membrane filter was used to filter the resultant hydrolysate, and phenylisothiocyanate was used for pre-column derivatization. A reversed-phase JASCO HPLC System (JASCO, Tokyo, Japan) fitted with a JASCO Unifinepak C18 (3.0 mm I.D. x 75 mm L, 1.9 \u0026micro;m) was used to perform the amino acid analysis. (JASCO, Tokyo, Japan HPLC-grade acetonitrile containing 0.1 M ammonium acetate and 0.1 M ammonium acetate at pH 6.5: They used methanol: water (46: 1 0: 44).. The flow rate was set at 1.0 mL/min, and the response of the UV detector was seen at 254 nm.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Gel Electrophoresis Analysis\u003c/h2\u003e \u003cp\u003eThe milk samples underwent a 10-minute, 12,000-g centrifugation at -4\u0026deg;C Next, the layer of fat was gently removed. After being collected from beneath the creamy layer, the milk serum was utilized for additional protein characterization procedures..The protein levels in milk samples were calculated using Lowry [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. Then, the protein samples were solubilized with phosphate buffer at specified concentrations and a temperature of 20\u0026deg;C. Subsequently, the solubilized samples underwent sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using a Bio-Rad System (Mini-Protean 3 cell, USA) following the method described by Laemmli [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Following electrophoresis, the gel was stained with Coomassie blue dye for two hours and then destained in a solution containing 30% methanol and 10% acetic acid. The molecular weight marker 10\u0026ndash;200 kDa (#SM0661) was from Fermentas (USA), and other reagents were of analytical grade.\u003c/p\u003e \u003cp\u003eThe Image Lab (Bio-Rad) software was used to examine the documentation's results using the densitometry technique. The software could predict molecular weight and relative density of each band which was relatively compared to bands from protein ladders/markers, so each molecular weight was determined. By calculating the standard protein quantity based on their density, the bands from BSA at varying concentrations were utilized to forecast the amount of each protein component. [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. Because the software can automatically create a regression from the standard band data, the quantity of the target band could be predicted. The collected data was then statistically examined to compare each component of goat and cow milk using the Mann-Whitney test.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Particle Size and Zeta Potential Analysis\u003c/h2\u003e \u003cp\u003eThe zeta potential and average particle size of raw goat milk were measured by diluting a 200 \u0026micro;L sample with 10 mL of 0.1 mol/L Na\u003csub\u003e2\u003c/sub\u003eHPO\u003csub\u003e4\u003c/sub\u003e-NaH\u003csub\u003e2\u003c/sub\u003ePO\u003csub\u003e4\u003c/sub\u003e (pH 7.0) buffer solution. To get rid of any dust and contaminants, the samples were then centrifuged at 1,078 \u0026times; g for five minutes at room temperature. Then, using the technique described by Li et al the protein particle size was examined using a HORIBA NANO PARTICA SZ-100 (Kyoto, Japan) nanometer particle size analyzer [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. A measurement temperature of 25\u0026deg;C, a scattering angle of 90\u0026deg;, a particle refractive index of 1.450, a particle absorptivity of 0.8872, and the usage of water as a dispersion with a dispersant refractive index of 1.330 were among the parameters considered in the analysis. Additionally, using a HORIBA NANO PARTICA SZ-100 (Kyoto, Japan) nanometer particle size analyzer with laser Doppler microelectrophoresis, the zeta potential of the goat milk samples was ascertained using the technique outlined by O'Brien et al [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.6 Color Analysis\u003c/h2\u003e \u003cp\u003eColor was measure on goat milk samples using the Chroma Meter CR-400 (Konica Minolta, Japan). The color readings were determined based on L* (Lightness), where L\u0026thinsp;=\u0026thinsp;100 for white and L\u0026thinsp;=\u0026thinsp;0 for black, Chroma a* (ranging from \u0026minus;\u0026thinsp;60 for green chromaticity to +\u0026thinsp;60 for red), and Chroma b* (ranging from \u0026minus;\u0026thinsp;60 for blue chromaticity to +\u0026thinsp;60 for yellow). It's important to note that the chroma meter was calibrated using a calibration plate.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e2.7 Determination of Rheological Properties\u003c/h2\u003e \u003cp\u003eA slightly modified Rheometer (MCR302, ANTON PAAR GMBH, Graz, Austria) was used to measure the rheological characteristics of goat milk. The sample tank was filled with 25 milliliters of goat's milk. A CC25 DIN/TI rotor and a CCB25 DIN/SS cup test head with a 120.00 mm diameter were used in the test. A scanning shear rate of 0.1 to 500 s-1 was used, and the clearance was fixed at 1.00 mm. Shear stress and apparent viscosity measurements were recorded for the entire test, which was carried out at 25\u0026deg;C. The consistency coefficient (K) and flow behavior index (n) were then measured by fitting the shear stress and shear rate curves.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e2.8 Confocal Microscopy Measurement\u003c/h2\u003e \u003cp\u003eFollowing Li and Shah's suggested methodology, the milk samples' structures were examined using the FV1200 confocal scanning laser microscope (Olympus) [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. The confocal scanning laser microscopy was outfitted with a silicon oil objective lens and an inverted microscope (magnification 150\u0026times;). A tagged image file format is used to acquire digital image files. Ten microliters of Nile red (1 mg/mL) were used to stain one milliliter of goat milk samples. in ethanol solution) and 10 \u0026micro;L of fluorescein isothiocyanate (FITC, 1 mg/mL in ethanol solution) for 30 min. Then, 20 \u0026micro;L of the stained material was pipetted onto a glass slide, covered with a coverslip, and put right into the confocal scanning laser microscope. In order to conduct the observations in a dark room, the emission wavelengths for FITC and Nile Red were set at 495 to 559 nm and 534 to 488 nm, respectively, and the excitation wavelengths were set at 500 to 600 nm.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e2.9 Statistical Analysis\u003c/h2\u003e \u003cp\u003eAnalysis of differences in milk quality between goat breeds was carried out using descriptive tests and one-way ANOVA using SPSS 23.0 software. Descriptive analysis includes the simplification and presentation of data so that the data that has been obtained in the field can provide preliminary information even though the information has not reached the stage of drawing conclusions. ANOVA was conducted to determine whether there was a difference in the diversity of the chemical quality of milk between the four goat breeds obtained. Using 30 variables chosen from Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, and four examples of goat breeds, principal component analysis (PCA) was used to separate groups belonging to various breeds.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe chemical quality of goat milk\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMilk Quality Paramater\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eKacang\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLocal Goat\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eEtawa Crossbreed\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eSaanen Crossbreed\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMoisture Content (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e85.48\u0026thinsp;\u0026plusmn;\u0026thinsp;2.14\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e86.86\u0026thinsp;\u0026plusmn;\u0026thinsp;0.39\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e87.76\u0026thinsp;\u0026plusmn;\u0026thinsp;1.84\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e87.52\u0026thinsp;\u0026plusmn;\u0026thinsp;0.98\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLactose (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.45\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.26\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFat (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eProtein (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.63\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.95\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAmino Acids (g/ 100 g of Protein)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAsp\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.15\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGlu\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e7.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e7.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSer\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.51\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGly\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.39\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.15\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHis\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.92\u0026thinsp;\u0026plusmn;\u0026thinsp;0.26\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.62\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eArg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.51\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.78\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eThr\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.87\u0026thinsp;\u0026plusmn;\u0026thinsp;0.76\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.45\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePro\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.56\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTyr\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.62\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.95\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCys\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.19\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMet\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIle\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.97\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLeu\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePhe\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLys\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.92\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.77\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003eNotes: Different superscripts on the same line indicate significant differences (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe physical quality of goat milk\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMilk Quality Paramater\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eKacang\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLocal Goat\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eEtawa Crossbreed\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eSaanen Crossbreed\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSpecific gravity\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.03320\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.03540\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.03340\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.03450\u0026thinsp;\u0026plusmn;\u0026thinsp;0.55\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eZeta Potential (mV)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-8.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-8.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-8.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-8.57\u0026thinsp;\u0026plusmn;\u0026thinsp;0.45\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eParticle Size (nm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e247\u0026thinsp;\u0026plusmn;\u0026thinsp;0.74\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e244\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e246\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e245\u0026thinsp;\u0026plusmn;\u0026thinsp;0.67\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003epH\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6.66\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.63\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eColor\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e83.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e83.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e86.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e85.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.67\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ea\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-1.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.83\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-1.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-2.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.41\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-1.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eb\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.56\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003eNotes: Different superscripts on the same line indicate significant differences (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05)\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Results and discussion","content":"\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e3.1 Physicochemical properties of goat milk\u003c/h2\u003e \u003cp\u003eThe physicochemical composition of raw milk is significantly influenced by the breed of the goat. Several noteworthy differences have been reported among four different breeds available in Gorontalo: kacang, local goat, ettawa crossbreed, and Saanen crossbreed (Tables\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe moisture content of goat milk showed minimal variations across different breeds, ranging from 85.32\u0026ndash;89.01% (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). This is noteworthy since moisture is a key factor in regulating the cost, microbiological stability, and quality of milk products (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).. The solution and colloidal suspension of additional milk ingredients are mediated by water. [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. The findings of this study align with those of Mohsin et al. (2019), who reported moisture contents in goat milk from British Alpine, Jamnapari, Saanen, Shami, and Toggenburg breeds as 87.29%, 85.32%, 88.08%, 88.33%, and 89.01% respectively. The Jamnapari breed exhibited a lower moisture content of 85.32% compared to other breeds such as Shami, Saanen, and Toggenburg [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Water acts as a mediator in the solution and colloidal suspension of other milk constituents [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e], [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eLactose content did not exhibit significant variation among the breeds (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05), ranging from 4.26\u0026ndash;4.55%. This uniformity suggests that lactose synthesis is largely conserved across breeds, consistent with findings by Park et al. (2007), who reported that genetic differences have a minimal effect on lactose concentration in goat milk [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. Lactose plays a critical role in milk osmoregulation, influencing the overall solute balance and milk yield [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. Furthermore, lactose levels in the four goat breeds studied were higher than the 3.69\u0026ndash;3.74% reported by Setiawan et al. [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. Lactose is the primary carbohydrate in milk, with goat milk containing 0.2\u0026ndash;0.5% less lactose than cow milk [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. Several factors influence lactose levels, including the nutrient composition of the feed given to livestock. Poor feed quality can result in lower lactose concentrations in milk[\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e] [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. In fermented milk, lactose provides lactic acid bacteria (LAB) with energy and is essential to generate acid during fermentation. [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe four goat breeds' milk had fat contents ranging from 4.08\u0026ndash;4.12%. According to the findings, the milk fat content of the four goat breeds was lower for Etawa goats (5.98\u0026ndash;6.98%) and Jamnapari/Etawa goats (4.61\u0026ndash;5.17%) [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e] [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. Fat content is influenced by several factors such as feeding types such as forage and concentrates. Providing forage will affect the formation of fat because forage is a source of fiber. The production of acetate significantly influences the synthesis of fatty acids, which in turn impacts the fat content in milk [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. When livestock consume forage, it undergoes a fermentative process in the rumen by rumen microbes, resulting in the formation of Volatile Fatty Acids (VFA) such as propionate, acetate, and butyrate. Acetate is then absorbed into the bloodstream and converted into fatty acids, which are subsequently transported into the secretory cells of the udder, contributing to the production of milk fat [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. It's important to note that both fat content and protein are key factors in determining the selling price of milk [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. Interestingly, all four goat milk products have consistent fat content (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05), underscoring the uniformity in this aspect of milk production.\u003c/p\u003e \u003cp\u003eProtein content, however, exhibited more pronounced variations (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05)., with Kacang and Local Goat breeds showing lower values (3.75% and 3.95%, respectively) compared to Etawa Crossbreed (4.11%) and Saanen Crossbreed (4.23%). The higher protein content in crossbreeds may be attributed to selective breeding practices aimed at improving milk yield and quality [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. Protein concentration is a crucial determinant of milk processing characteristics, particularly in cheese production, where higher protein content enhances curd formation and yield [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. The protein content in milk is directly influenced by the protein content in the feed, with higher dietary protein levels resulting in increased milk protein secretion. The primary source of protein in animal feed typically comes from concentrates [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. Enhancing the availability of amino acids in feed has been shown to promote milk protein synthesis [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. The production of milk protein starts when animals eat concentrate-based diet, which is then converted into amino acids by rumen bacteria. These amino acids are then absorbed in the small intestine, transported into the bloodstream, and eventually delivered to the secretory cells of the udder, where they contribute to milk protein synthesis [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eSpecific gravity serves as a key indicator of milk composition, reflecting the concentration of dissolved solids such as proteins, fats, and lactose. The recorded values ranged from 1.03320 to 1.03540, with the highest specific gravity observed in Local Goat milk (1.03540\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09) and the lowest in Kacang goat milk (1.03320\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11). However, statistical analysis indicated that these differences were not significant (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05), suggesting that all breeds exhibit relatively similar concentrations of milk solids. The specific gravity of milk from kacang goat is 1.030 [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e], besides the specific gravity of milk from etawa crossbreed 1.030 and saanen crossbreed 1.0295 [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. This could be influenced by the fat content of milk which has a negative impact on the specific gravity of milk [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]. According to McCarthy and Singh [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e] stated that the specific gravity of milk depends on the fat content and solid material of the milk, because the specific gravity of fat is lower than the specific gravity of density of water or milk plasma. The release of gases such as carbon dioxide (CO\u003csub\u003e2\u003c/sub\u003e) and nitrogen (N\u003csub\u003e2\u003c/sub\u003e) into milk immediately following the milking process can lead to an increase in specific gravity. Understanding this phenomenon is essential for monitoring milk quality and ensuring optimal processing conditions [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. Goat milk has a specific gravity that is lower than sheep milk (range: 1.0347\u0026ndash;1.0384 kg/m\u003csup\u003e3\u003c/sup\u003e) but higher than cow's milk (range: 1.0231\u0026ndash;1.0398 kg/m\u003csup\u003e3\u003c/sup\u003e) [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThere was no discernible difference in the pH of the milk from the four goat breeds (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05). Milk from various goat breeds has a pH value that ranges from 6.61 to 6.65, which suggests possible milk damage [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e]. The precise pH value is influenced by components present in fresh milk, such as CO\u003csub\u003e2\u003c/sub\u003e, phosphate, citrate, and protein, which affect the milk's ability to resist changes in pH and acidity, thus preventing milk spoilage [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e]. Bacterial activity can cause the pH to become more acidic, dropping below the average value of 6.5\u0026ndash;6.7 [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. pH values higher than 6.7 usually indicate the possibility of mastitis [\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe colour characteristics of goat milk from four genotypes\u0026mdash;Kacang, Local Goat, Etawa Crossbreed, and Saanen Crossbreed\u0026mdash;were evaluated using the CIE L\u003cem\u003ea\u003c/em\u003eb* system, where L* denotes lightness, a* indicates the green\u0026ndash;red axis, and b* represents the blue\u0026ndash;yellow axis. Among the genotypes, Etawa Crossbreed milk exhibited the highest L* value (86.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13), indicating a significantly lighter appearance compared to the other groups (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05), followed by Saanen Crossbreed (85.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.67). These findings suggest a superior visual brightness, which is often associated with higher consumer appeal and milk freshness [\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e]. In contrast, Kacang and Local Goat milk showed lower lightness values (both 83.2\u0026ndash;83.3), indicating a comparatively darker hue. The higher L* values in goat milk are related to smaller fat globule size and more complete conversion of β-carotene into colorless vitamin A, resulting in a whiter appearance [\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]. This whitening effect enhances light scattering, making crossbred goat milk visually more appealing to consumers. The a* values across all genotypes were negative, ranging from \u0026minus;\u0026thinsp;1.49 to \u0026minus;\u0026thinsp;2.10, which indicates a slight shift toward the green spectrum. While these differences were not statistically significant (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05), they align with the findings of Milovanovic et al [\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e], who reported higher greenish tones in goat and sheep milk compared to cow milk. The greenish hue is attributed to the lack of residual carotenoids, a trait unique to goat species due to their metabolic conversion pathway [\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e][\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]. Regarding b* values, all breeds showed mild yellowish tones (b* = 4.81\u0026ndash;5.45), with Etawa Crossbreed having the highest yellow component. The yellow appearance may be attributed to residual carotenoids or fat-soluble pigments, although values were lower than typically seen in cow milk, consistent with literature noting the lack of β-carotene conversion in goat milk [\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e]. Overall, the colour traits observed reinforce the role of genotype in influencing milk visual attributes, with crossbred goats (especially Etawa) producing milk that appears brighter and slightly more yellow\u0026mdash;features that may enhance its marketability and consumer preference, as confirmed by both instrumental and visual assessments in Milovanovic et al [\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e3.2 Amino acid and protein profile\u003c/h2\u003e \u003cp\u003e The protein composition of goat milk from several breeds in Gorontalo, including Kacang, Local Goat, Etawa Crossbreed, and Saanen Crossbreed, was displayed in the SDS-PAGE (Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis) image (Fig.\u0026nbsp;1). The molecular weight distribution and relative abundance of casein and whey protein fractions, which are intimately related to the goat milk's amino acid composition, can be learned from the SDS-PAGE analysis of goat milk proteins (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Breed-specific differences in protein expression are confirmed by the protein bands seen in the SDS-PAGE gel, especially in β-casein, α-casein, and κ-casein (25\u0026ndash;35 kDa), as well as β-lactoglobulin and β-lactalbumin (14\u0026ndash;18.4 kDa). These variations are consistent with the amino acid profiles found in the milk of the Saanen, Etawa, Kacang, and Local goat breeds..\u003c/p\u003e \u003cp\u003eThe primary protein component of goat milk, caseins are necessary for both the stability of the milk and the creation of cheese [\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e]. The Fig.\u0026nbsp; 1 results show that Etawa and Saanen Crossbreeds exhibit more intense casein bands, indicating higher casein concentrations compared to Kacang and Local Goat breeds. This observation correlates with the higher glutamic acid (Glu) and aspartic acid (Asp) concentrations in these breeds, as shown in the amino acid composition (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Aspartic acid (Etawa: 3.14 g/100 g, Saanen: 2.15 g/100 g) along with glutamic acid (Etawa: 7.02 g/100 g, Saanen: 7.14 g/100 g) are known for their role in stabilizing protein structures and enhancing the emulsification properties of milk [\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e]. Additionally, higher levels of proline (Pro) and threonine (Thr) in Etawa and Saanen Crossbreeds further support their superior casein content [\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e]. Proline (Etawa: 5.02 g/100 g, Saanen: 3.11 g/100 g) enhances protein stability, while threonine (Etawa: 6.24 g/100 g, Saanen: 4.11 g/100 g) is essential for protein synthesis and milk viscosity, aligning with the observed rheological differences among the breeds.\u003c/p\u003e \u003cp\u003eWhey proteins, particularly β-lactoglobulin and β-lactalbumin, play a significant role in immune function, digestibility, and processing properties [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e]. Etawa and Saanen Crossbreeds have stronger whey protein bands, according to Fig.\u0026nbsp;1 data, which is in keeping with their greater levels of branched-chain amino acids (BCAAs), including valine (Val), isoleucine (Ile), and leucine (Leu). These amino acids are critical for protein metabolism and muscle development, making the milk from these breeds more suitable for high-protein formulations and sports nutrition products [\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e]. Specifically, Etawa Crossbreed showed the highest levels of leucine (2.44 g/100 g), isoleucine (2.12 g/100 g), and valine (3.14 g/100 g), aligning with the observed abundance of β-lactoglobulin in its SDS-PAGE profile. Saanen Crossbreed also demonstrated high levels of these amino acids, reinforcing its potential for nutritional applications, particularly in infant formula and dietary supplements [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003e3.3 Extended protein fraction estimation in goat milk from four genotypes\u003c/h2\u003e \u003cp\u003eProtein composition is one of the most critical determinants of milk quality, influencing its nutritional value, processing behavior, and functional applications. The main protein fractions\u0026mdash;β-casein, α-casein, κ-casein, β-lactoglobulin, and β-lactoalbumin\u0026mdash;were estimated in this work using SDS-PAGE electrophoresis (Fig.\u0026nbsp; 1) and densitometric analysis in goat milk obtained from four genotypes: Kacang, Local Goat, Etawa Cross, and Saanen Cross. These estimations were calculated using densitometric band intensity data ((in arbitrary units, A.U.)) and translated into approximate concentrations (g/L), Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e following a conversion approach similar to that of Li et al [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e], which assumes a linear relationship between band intensity and protein concentration under Coomassie Brilliant Blue staining.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eProtein Fraction of Goat Milk\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGenotype\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eβ-Casein (g/ L)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eα-Casein (g/ L)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eκ-Casein (g/ L)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eβ-Lactoglobulin (g/ L)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eβ-Lactalbumin (g/ L)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eKacang\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.98\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.43\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.56\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLocal Goat\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.45\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEtawa Crossbreed\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.78\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSaanen Crossbreed\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.62\u0026thinsp;\u0026plusmn;\u0026thinsp;0.76\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.91\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"6\"\u003eNotes: Different superscripts on the same column indicate significant differences (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05)\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eAmong the genotypes evaluated, Saanen Cross demonstrated the highest total protein concentration (9.05 g/L), primarily due to elevated casein levels. This group showed 5.6 g/L of β-casein, alongside estimated concentrations of 1.96 g/L of α-casein and 1.4 g/L of κ-casein, making it the most casein-rich milk type in this study. The dominance of β-casein, comprising over 60% of the total protein, aligns with findings from Costa et al [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e], who also reported Saanen milk to contain high levels of β- and αs1-caseins. These fractions are essential for curd formation, texture development, and yield in cheese-making processes [\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e]. Conversely, Kacang goat milk presented a markedly different profile, with a total protein content of 7.1 g/L, of which β-lactoglobulin accounted for 4.3 g/L\u0026mdash;the highest among all breeds. Its estimated β-casein concentration was 2.8 g/L, with accompanying levels of 0.98 g/L α-casein and 0.7 g/L κ-casein. This composition reflects a whey-dominant profile, suitable for applications in specialized nutrition, including infant formula or medical nutrition products due to higher digestibility and lower allergenicity [\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e]. The relatively high β-lactoalbumin content (1.29 g/L) further enhances this nutritional suitability, given its known role in lactose synthesis and immune functions. Etawa Cross and Local Goat displayed intermediate profiles. Etawa Cross milk contained 5.1 g/L β-casein, with corresponding values of 1.79 g/L α-casein and 1.28 g/L κ-casein. Local Goat milk showed 4.1 g/L β-casein, 1.44 g/L α-casein, and 1.03 g/L κ-casein. Both genotypes had balanced whey and casein fractions, making them versatile for both processing and dietary formulations [\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e]. Notably, their protein-to-casein ratios and viscosity values positioned them between the Kacang and Saanen genotypes, suggesting multifaceted industrial potential.\u003c/p\u003e \u003cp\u003eOverall, the extended fractionation of goat milk proteins provides a nuanced understanding of compositional diversity among breeds. The casein-to-whey ratio, along with individual casein subtype estimates, directly informs processing decisions. Higher casein levels favor curd firmness and yield, while elevated whey protein content enhances biofunctionality. These insights underscore the importance of genotype selection in dairy breeding programs and the design of targeted dairy products for specific nutritional and technological purposes.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003e3.4 Zeta potential and particle size\u003c/h2\u003e \u003cp\u003eZeta potential is an important measure for understanding the stability and charge of milk systems. This study looked at the zeta potential values in goat milk from four types of goats: Kacang, Local Goat, Etawa Crossbreed, and Saanen Crossbreed. The values ranged from \u0026minus;\u0026thinsp;8.48 to \u0026minus;\u0026thinsp;8.65 mV (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The Etawa Crossbreed had the most negative value at \u0026minus;\u0026thinsp;8.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33 mV, suggesting better stability due to stronger electrostatic repulsion. The lowest negative value was \u0026minus;\u0026thinsp;8.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22 mV for Kacang goat milk, although the differences were not statistically significant (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05). These findings match earlier research by Li et al [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e], which found raw goat milk zeta potential values between \u0026minus;\u0026thinsp;8.50 and \u0026minus;\u0026thinsp;8.75 mV. However, our results differ from Huo et al [\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e], who reported a value of \u0026minus;\u0026thinsp;27.5 mV for Aote Laoshan goat milk, and Chen et al [\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e], who found a value of \u0026minus;\u0026thinsp;30.3 mV for raw goat milk. The difference in zeta potential may be because of variations in milk composition related to different goat breeds, especially in proteins and fats. The charge of the micellar particles in milk is influenced by the casein and whey proteins. Glutamic and aspartic acids are among the many negatively charged amino acids found in caseins, especially β-casein and αs1-casein. Past research indicates that crossbred goats, such as Saanen and Etawa, usually produce milk with more casein and richer in these acids, which may lead to more negative zeta potential values [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Additionally, differences in the milk fat globule membrane (MFGM) and the size of fat globules may also affect the surface charge. Goats that are crossbred typically produce milk with smaller fat globules, increasing the milk's surface area.. This exposes more charged phospholipids and glycoproteins on the MFGM, increasing electrostatic repulsion. In contrast, goats may have larger fat globules with different membrane compositions, resulting in a reduced surface charge and a less negative zeta potential [\u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e]. Mineral content, especially calcium and phosphate ions, can also affect the surface charge because they bind to casein micelles. While generally minor, differences in ionic composition among goat breeds can lead to small variations in their behavior in an electric field [\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eGoat milk samples' particle size distribution was also assessed in order to identify structural traits that are important for stability and digestion. Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e explain the particle diameters ranged narrowly between 244 and 247 nm across the four genotypes, with no significant statistical differences (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05), indicating a relatively consistent colloidal system regardless of genotype. The size range and distribution properties of the particles in the protein solution are reflected in the particle size distribution. Raw goat milk protein has a wide and irregular bimodal particle size distribution, with most of the protein particles localized around 200 nm, as demonstrated by Huo et al. [\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e]. Comparative data from Li et al [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e] support this, showing that raw goat milk averaged 250 nm. Because different breeds differ in their milk composition, especially in terms of fat and protein content, as well as their molecular structures, which are impacted by breed-specific genetics, food, and lactation physiology, the particle size of raw goat milk can also vary [\u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e]. Protein interaction and fat globule size: Goat milk is easier to digest than cow milk because it naturally contains fewer fat globules and less αS1-casein. [\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e]. However, the size and uniformity of fat globules and protein aggregates still vary by breed due to inherent genetic and biochemical differences. For example, differences in casein micelle structure and whey protein concentration between breeds affect aggregation and emulsification behavior, leading to different particle size distributions​. Breed influence on milk composition; goat milk composition\u0026mdash;including the types and amounts of caseins and whey proteins\u0026mdash;differs between breeds. These compositional differences affect thermal stability and response to processing, ultimately influencing particle size and emulsion stability​. The particle size distribution (e.g., bimodal vs. uniform) and changes in zeta potential indicate breed-dependent responses in milk microstructure when subjected to processing. Ultrasonic treatment, for instance, results in more uniform and smaller particles, but the starting size and distribution in raw milk still reflect breed-specific characteristics​. Thus, differences in particle size in raw goat milk from various breeds arise primarily due to variations in fat globule size, protein content and structure, and overall milk composition, all of which are inherently linked to the genetic traits of each goat breed.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003e3.5 Rheological behavior of goat milk\u003c/h2\u003e \u003cp\u003eShear rate is a fundamental parameter in rheological analysis that describes the deformation rate of a fluid under applied stress. In dairy science, understanding the shear rate behavior of goat\u0026rsquo;s milk is critical for optimizing its processing, stability, and texture in various dairy applications. Goat\u0026rsquo;s milk, like other dairy fluids, can exhibit non-Newtonian flow behavior, depending on its composition. Figure\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003e shows a non-linear, goat\u0026rsquo;s milk exhibits non- Because of the existence of fat globules, casein micelles, and protein structures, Newtonian features are more prevalent [\u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e] [\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e]. The viscosity decreases as shear rate increases, which is a key property of goat\u0026rsquo;s milk due to its colloidal nature. At high shear rates; a declining viscosity trend with increasing shear rate would confirm pseudoplastic behavior, meaning the milk becomes thinner as shear increases. This is a desirable property in dairy processing, making milk easier to pump, mix, and homogenize without phase separation. The shear rate response of goat\u0026rsquo;s milk is influenced by breed differences: Etawa and Saanen Crossbreeds typically show higher shear resistance due to increased protein and total solids content, resulting in a more structured milk network. Kacang and Local Goat breeds tend to have lower viscosity and faster shear response, indicating a more fluid-like consistency with smaller fat globules and lower solid content. Implications for dairy processing and stability; understanding shear rate behavior ensures optimal homogenization conditions, preventing fat separation in processed milk. Shear-thinning properties are beneficial for products like yogurt and cheese, where controlled viscosity is required for desirable texture and mouthfeel. Milk with lower shear resistance may experience phase separation faster, requiring stabilization techniques for extended shelf life.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eViscosity is a crucial rheological parameter that determines the flow behavior and textural properties of goat\u0026rsquo;s milk, influencing its processing, stability, and sensory perception. The analysis of viscosity in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003e provides insights into the physical characteristics of goat\u0026rsquo;s milk in Gorontalo, which are affected by breed composition, fat and protein content, and processing conditions. In general, goat's milk has non-Newtonian fluid behavior, meaning that as the shear rate increases, its viscosity falls, suggesting pseudoplastic (shear-thinning) qualities [\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e]. This is characteristic of milk due to the presence of casein micelles and fat globules that align under shear stress, reducing resistance to flow. The data in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003e show a declining viscosity trend with increasing shear rate, it confirms the pseudoplastic nature of goat\u0026rsquo;s milk, similar to previous studies on dairy rheology. Because goat breeds range in terms of fat globule size and protein concentration, the viscosity of their milk can vary greatly, and total solids content. Typically; higher viscosity is associated with higher protein and fat content, as seen in breeds such as Etawa and Saanen Crossbreeds. Lower viscosity is observed in milk with lower total solids and smaller fat globules, which is characteristic of Kacang and Local Goat breeds. Figure\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003e demonstrates variations among breeds, it suggests that compositional differences play a major role in determining viscosity. Milk viscosity is highly temperature-dependent, with lower temperatures increasing viscosity due to reduced molecular mobility [\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e]. It is expected that higher temperatures result in decreased viscosity, following the Arrhenius equation for fluid dynamics [\u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e]. This is essential for optimizing processing conditions such as pasteurization, homogenization, and ultrafiltration in dairy industries.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003e3.6 Microstructure analysis\u003c/h2\u003e \u003cp\u003e The microstructural features of goat milk from several breeds in Gorontalo, including Kacang, Local Goat, Etawa Crossbreed, and Saanen Crossbreed, are depicted in the fluorescence microscope photos above (Fig.\u0026nbsp;4).This analysis provides insights into the fat globule distribution, protein network, and overall structural differences, which directly influence milk stability, digestibility, and dairy processing potential. The size and distribution of fat globules play a critical role in determining milk texture, mouthfeel, and processing behavior [\u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e72\u003c/span\u003e]. Kacang and Local Goat milk show a denser distribution of smaller fat globules, suggesting better emulsion stability and easier digestibility, which aligns with findings that smaller globules improve lipid absorption [\u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e73\u003c/span\u003e]. Etawa and Saanen Crossbreed milk exhibit larger and more heterogeneous fat globules, indicating a higher fat content.\u003c/p\u003e \u003cp\u003eImplications for dairy processing; higher fat globule uniformity in Kacang and Local Goat milk makes it better suited for fluid milk consumption and products requiring emulsion stability, such as yogurt and infant formula. Larger fat globules in Etawa and Saanen Crossbreed milk contribute to richer texture and creamier consistency, making them ideal for cheese production, butter, and high-fat dairy products. The differences in fat globule structure among breeds highlight the need for specific processing techniques, such as homogenization, pasteurization, and fat standardization, to optimize dairy product quality.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003e3.7 Principal component analysis (PCA)\u003c/h2\u003e \u003cp\u003eThe PCA biplot provides a comprehensive visual representation of how different breeds of goats are distributed based on their physical milk composition. It illustrates the relationships among the measured variables and how they contribute to the overall variation in physical milkFigure 5A displays the plot of the PCA model of physical milk composition, while Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e5\u003c/span\u003eB displays the plot of the PCA model of chemical milk composition.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eEighty-two percent of the variations in the data set were displayed by the first two PCs (PC1 and PC2) (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e5\u003c/span\u003eA). The PC1 was most affected by the density (specific gravity) and brightness (L) of the milk, accounting for 52.94%.. The PC2 accounted for 29.06% of the captures variations in Zeta Potential, pH, and minor color properties. The analysis of the samples in quadrant I (PC1+, PC2+) showcases the outstanding qualities of milk from the Etawa crossbreed. With the highest L values indicating brightness and a larger particle size, this breed emerges as one of the most favorable options for dairy processing, underscoring its potential to make a significant contribution to the industry. In quadrant II (PC1-, PC2+), we find valuable insights from the milk of the Kacang goat. This breed plays a notable role in the overall variation in milk properties, characterized by a higher specific gravity and lower lightness (L), along with a distinctive Zeta Potential that reflects the electrical charge on its milk particles. These characteristics suggest that Kacang goat milk may be particularly well-suited for concentrated dairy products, highlighting the breed's importance in specialized dairy applications. Quadrant III (PC1-, PC2-) includes milk from the Local goat, which exhibits lower L* values and potentially different specific gravity or a* values. This indicates that the milk from the Local goat is versatile and well-suited for general dairy applications, including pasteurized and fresh milk, thereby enabling a broader range of dairy products. Finally, the samples in quadrant IV (PC1+, PC2-) present milk from the Saanen breed. While it shares similarities with Etawa milk, it displays slightly lower brightness and a moderate particle size. Saanen milk offers a robust option for standard dairy applications that demand high-quality and stable products. By understanding these unique properties, producers can make informed decisions that effectively enhance their dairy offerings.\u003c/p\u003e \u003cp\u003ePlotting of the PCA model of chemical milk composition is displayed in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e5\u003c/span\u003eB.. 92.71% of the variations in the data set were displayed by the first two PCs (PC1 and PC2). Moisture content, lactose, fat, and protein composition were the main factors influencing the PC1, which explained 78.73% of the overall variance.. With the exception of moisture content, which was found at PC1's negative loading, all of these variables showed positive correlations. The amounts of amino acids (Asp, Glu, Thr, Val, etc.) were the main cause of the PC2's 13.98% variance. Known for its outstanding quality, milk from the Etawa crossbreed is highlighted in Quadrant I (PC1+, PC2+).. This quadrant is defined by high moisture content, elevated protein levels, and a rich array of essential amino acids, including glutamic acid (Glu), aspartic acid (Asp), and branched-chain amino acids like leucine, isoleucine, and valine. The strong correlation between increased protein and these amino acids indicates that Etawa milk is highly nutritious, making it suitable for functional dairy products such as fortified yogurts and premium cheeses. Quadrant II (PC1-, PC2+) presents Kacang goat milk, characterized by lower moisture and protein content but higher fat concentration and amino acids like cysteine (Cys) and methionine (Met). The inverse relationship between these amino acids and moisture suggests that Kacang milk has a higher dry matter concentration, making it particularly advantageous for rich cheeses and butter. Quadrant III (PC1-, PC2-) features milk from local goats, which, while lower in protein and amino acids, has moderate moisture and lactose levels. It remains valuable for fresh goat milk products and traditional fermented items, benefiting from a mild flavor. Quadrant IV (PC1+, PC2-) showcases Saanen crossbreed milk, which shares beneficial traits with the Etawa breed. Though it has slightly lower concentrations of select amino acids, it retains high moisture and protein content, making it suitable for high-nutritional-value dairy products, including premium cheeses and quality cream. Dairy producers may strategically maximize milk utilization, improve product quality, and satisfy changing market demands by acknowledging the distinctive qualities of each breed.\u003c/p\u003e \u003cp\u003eTo improve our understanding of milk quality and its market potential, future research should focus on several key areas. First, investigating the impact of dietary composition on the physicochemical properties of milk could optimize nutrient intake and enhance product stability. Additionally, assessing the influence of various lactation stages on milk composition through Principal Component Analysis (PCA) would provide valuable insights into the dynamic changes in milk quality over time. Furthermore, exploring consumer preferences regarding goat milk characteristics could help align dairy product development with market demand. Future studies should also examine the combined effects of diet and lactation stage on the chemical composition of milk. Adapting processing methods to accommodate breed-specific variations in milk properties may further improve both its nutritional value and commercial viability. Moreover, establishing market segmentation strategies based on milk composition could optimize product quality and maximize economic benefits in the dairy industry.\u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003e This study presents a comprehensive characterization of the physicochemical and rheological properties of raw goat milk from four genotypes\u0026mdash;Kacang, Local Goat, Etawa Crossbreed, and Saanen Crossbreed\u0026mdash;reared in Indonesia. The results demonstrated that genotype significantly influences milk composition, particularly in protein content, amino acid profiles, micellar protein structure, and rheological behavior. Crossbreeds, notably Saanen and Etawa, produced milk with higher total protein and casein fractions, contributing to enhanced viscosity and suitability for cheese and fermented dairy production. Conversely, Kacang goat milk exhibited higher whey protein content and sulfur-rich amino acids, aligning with applications in specialized nutrition. Despite breed-dependent differences in microstructure, all milk types displayed consistent pseudoplastic behavior and colloidal stability, as confirmed by zeta potential and particle size measurements. These findings highlight the functional diversity of goat milk in Indonesia and support targeted valorization strategies for each breed. This study enriches the existing knowledge base by providing molecular and rheological insights that can guide breed selection, dairy processing optimization, and functional food development. Future research should investigate the effects of feed and lactation stage on milk quality and explore consumer preferences to bridge scientific potential with market viability.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e Funding by the Research Institute for Humanity and Nature (RIHN) Japan, No 14200102.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e Data is provided within the manuscript or supplementary information files\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical trial registration\u0026nbsp;\u003c/strong\u003eNot applicable\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e Permission to conduct this research was obtained from the Faculty Board of the Faculty of Agriculture, State University of Gorontalo, in accordance with the standard operating procedures established by the institution\u0026rsquo;s Research and Development Committee. This study did not involve the use or handling of live animals, including breeding or maintenance within an animal facility. The procedures for the collection, processing, and utilization of milk materials in this study adhered to all applicable local, national, and international ethical guidelines for the use of animal-derived products in scientific research.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e Not applicable.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e The authors declare no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eJ. Sumarmono, \u0026lsquo;Current goat milk production, characteristics, and utilization in Indonesia\u0026rsquo;, \u003cem\u003eIOP Conf Ser Earth Environ Sci\u003c/em\u003e, vol. 1041, no. 1, p. 012082, Jun. 2022, doi: 10.1088/1755-1315/1041/1/012082.\u003c/li\u003e\n\u003cli\u003eQ. H. ALKaisy, J. S. 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Wang \u003cem\u003eet al.\u003c/em\u003e, \u0026lsquo;Size-dependent composition and in-situ structure analysis of the milk fat globule membrane in buffalo milk\u0026rsquo;, \u003cem\u003eFood Chem\u003c/em\u003e, vol. 464, p. 141766, Feb. 2025, doi: 10.1016/J.FOODCHEM.2024.141766.\u003c/li\u003e\n\u003c/ol\u003e\n"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"discover-food","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"discoverfood","sideBox":"Learn more about [Discover Food](https://www.springer.com/44187)","snPcode":"","submissionUrl":"","title":"Discover Food","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Physicochemical, Rheological, Milk, Local Goats, Gorontalo","lastPublishedDoi":"10.21203/rs.3.rs-6429174/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6429174/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe physicochemical and rheological characteristics of goat milk are pivotal in determining its processing potential and nutritional value. Despite Indonesia\u0026rsquo;s rich diversity in goat breeds, scientific data comparing the milk quality across these breeds remain scarce. This study investigated the physicochemical, rheological, structural, and protein characteristics of raw goat milk from four genotypes\u0026mdash;Kacang, Local Goat, Etawa Crossbreed, and Saanen Crossbreed\u0026mdash;reared under traditional farming conditions in Gorontalo, Indonesia. The aim was to identify breed-specific differences that may influence milk\u0026rsquo;s nutritional quality and processing suitability. A total of 48 raw milk samples were analyzed using standard compositional assays, sodium dodecyl sulfate\u0026ndash;polyacrylamide gel electrophoresis, densitometry, amino acid profiling, laser confocal microscopy, rotational rheometry, and colloidal stability measurements. These methods provided detailed insights into protein structure, micelle behavior, and textural properties. The results revealed that crossbred goats, particularly Saanen and Etawa, produced milk with higher protein content, stronger casein expression, greater viscosity, and richer profiles of glutamic acid, proline, and threonine. Conversely, Kacang goats showed higher whey protein content and elevated sulfur amino acids, such as cysteine and methionine. All samples exhibited pseudoplastic flow behavior and comparable zeta potential and particle size, indicating similar colloidal stability. These findings demonstrate the influence of genotype on goat milk functionality and support its valorization in developing sustainable and differentiated dairy products with enhanced nutritional and technological properties.\u003c/p\u003e \u003cp\u003e.\u003c/p\u003e","manuscriptTitle":"Uncovering the Functional Potential of Goat Milk: Physicochemical and Rheological Comparison Across Local Goats in Gorontalo Indonesia","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-09 09:12:24","doi":"10.21203/rs.3.rs-6429174/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-06-22T07:08:51+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-06-19T12:53:58+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"85328320139151402471821269851261452800","date":"2025-06-19T03:16:35+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"335513104653291519027083814633671005220","date":"2025-06-15T12:41:22+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-06-14T18:13:19+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"194103700910276618763472388040514527189","date":"2025-06-12T17:55:55+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"304396062319575735281397845973364585302","date":"2025-06-05T18:47:24+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-06-05T17:46:43+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-06-01T04:48:31+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-05-21T04:30:08+00:00","index":"","fulltext":""},{"type":"submitted","content":"Discover Food","date":"2025-05-21T04:29:04+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"discover-food","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"discoverfood","sideBox":"Learn more about [Discover Food](https://www.springer.com/44187)","snPcode":"","submissionUrl":"","title":"Discover Food","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"d2b83704-17b8-42ef-b9c6-a2beb73deaa9","owner":[],"postedDate":"June 9th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2025-11-13T09:53:10+00:00","versionOfRecord":[],"versionCreatedAt":"2025-06-09 09:12:24","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6429174","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6429174","identity":"rs-6429174","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}