{"paper_id":"0304c30a-ea12-484f-8a36-1d94c7a31540","body_text":"Effect of toxin binder feeding on body weight, milk and components yield, and aflatoxin M1 excretion in crossbred lactating cows | 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 Effect of toxin binder feeding on body weight, milk and components yield, and aflatoxin M1 excretion in crossbred lactating cows Himansu Agrawal, Bharat R. Devalia, Akash M. Sutariya, Jigar H. Patel, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9039431/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 6 You are reading this latest preprint version Abstract Mycotoxin contamination of livestock feed poses a significant threat to animal health, productivity, and food safety. To overcome this, study was conducted to evaluate the effect of toxin binder supplementation on body weight, body condition score, milk yield and composition, and aflatoxin M1 (AFM1) excretion in milk of crossbred lactating cows. Eighteen multiparous cows (75% Holstein Friesian × 25% Kankrej- Indian Zebu), similar in milk yield, days in milk, and lactation numbers were randomly divided into three groups of six each and fed for 70 days. The control group (T1) fed basal total mixed ration (TMR) without toxin binder, and two supplemented groups receiving toxin binder at 50 g/day (T2) and 100 g/day (T3). The toxin binder comprised hydrated sodium calcium aluminosilicate (bentonite), organic acids, mannan oligosaccharides, Bacillus (direct-fed microbial), oxine copper, and herbal ingredients. Toxin binder supplementation improved (p < 0.001) milk yield, 4% fat-corrected milk, and energy-corrected milk yields, with T2 showing the highest increase. Milk composition was maintained in T2 but declined at higher dosage (T3). Despite greater aflatoxin B1 intake with higher feed consumption, toxin binder markedly reduced AFM1 concentration and excretion in milk by 76.8–78.7% and 73.2–75.2%, respectively, compared with control, and lowered bio-concentration and carry-over percentages by about 75–78%. The toxin binder also not influenced liver, kidney function and energy-protein nutrition. The treatment cost per kilogram of milk also decreased by 97.2% in T2 group. It was concluded that supplementation of toxin binder at 50 g/day was most effective in improving milk and components yield, and welfare of cow while minimizing aflatoxin health risk to consumers. Aflatoxin M1 Body weight Crossbred cows Feed safety Milk yield Toxin binder Figures Figure 1 INTRODUCTION Mycotoxin contamination of livestock feed poses a significant threat to animal health, productivity, and food safety. Dairy animals consume a wide variety of feedstuffs, including concentrates, silages, and forages, which are vulnerable to fungal contamination. Aflatoxins, produced by Aspergillus flavus, A. parasiticus , and A. nomius , are the most harmful mycotoxins (Gonçalves et al., 2017 ). Aflatoxin B1 (AFB1) is the most potent, classified by the International Agency for Research on Cancer as a Group I human carcinogen (IARC, 2002 ). In ruminants, AFB1 is partly degraded in the rumen (Tolosa et al., 2021 ), but the absorbed fraction undergoes hepatic metabolism, generating toxic intermediates that damage DNA, proteins, and lipids (Cupid et al., 2004 ). In dairy cows, AFB1 is rapidly bio-transformed into aflatoxin M1 (AFM1), which appears in milk within hours of ingestion (Guo et al., 2021 ; Gallo et al., 2008 ; Battacone et al., 2003 ). Carry-over rates range from 1% to 6%, depending on milk yield, breed, and stage of lactation (Campagnollo et al., 2016 ; Battacone et al., 2003 ; Veldman et al., 1992 ). The impact of mycotoxin on hematological and biochemical indices is also limited. Several strategies have been explored to mitigate the excretion of aflatoxin in milk. Among them, toxin binder supplementation remains the effective and practical on-farm approach. Adsorbents such as bentonite, montmorillonite, hydrated sodium calcium aluminosilicate (HSCAS), zeolites, activated charcoal, and yeast cell wall components are widely used (Čolović et al., 2019 ). Among these, HSCAS has demonstrated significant effectiveness, binding up to 95% of aflatoxins and nearly halving AFM1 excretion in milk (Queiroz et al., 2012 ; Kutz et al., 2009 ). At the same time, biological detoxification using microorganisms such as Bacillus subtilis, Rhodococcus erythropolis , and Pleurotus pulmonarius has shown promise in degrading aflatoxins, although further in vivo validation is required (Loi et al., 2016 ; Eshelli et al., 2015 ; Gao et al., 2011 ). The toxin binder evaluated in this study consisted of bentonite, mannan oligosaccharides (MOS), Bacillus (direct-fed microbial), organic acids, oxine copper, and herbal extracts, designed to provide complementary benefits, including toxin adsorption, microbial detoxification, immune modulation, gut environment stabilization, and hepato-protective effects. The study was conducted to evaluate the effects of toxin binder supplementation on body weight, body condition score, milk and components yield as well as on aflatoxin excretion in milk of lactating crossbred cows. MATERIALS AND METHODS The experiment was conducted for 70 days, following a ten-day adaptive feeding period. A total of eighteen 75% Holstein Friesian x 25% Kankrej- Indian Zebu crossbred lactating cows were randomly allotted to 3 treatment groups (6 in each) based on similarity of milk yield (19.09 ± 0.21 kg/day), days in milk (71.8 ± 14.3 days), and lactation numbers (2.11 ± 0.19). The cows of three treatment groups viz . Control (T1), Treatment 2 (T2), and Treatment 3 (T3) were fed ad-libitum total mixed ration (TMR). The composite toxin binder containing hydrated sodium calcium alumino-silicate (bentonite), organic acids, mannan oligosaccharides, Bacillus direct-fed microbial, oxine copper, and herbal ingredients fed to cows of T2 and T3 groups at 50 g/day and 100 g/day, respectively. The toxin binder fed to cows in equal half after mixing with 250 g compounded concentrate mixture (CCM) in morning and evening in treatment groups and equal quantity of CCM also fed to cows of control group. The CCM kept open before feeding, AFM1 excretion in milk before and after feeding were measured for this experiment. The TMR was prepared using 42kg compounded concentrate mixture, 13kg wheat straw, 24kg groundnut straw, 15kg hybrid Napier, 4kg bypass fat, 1kg mineral-vitamin premix, and 1kg salt. All the crossbred cows were housed in well-ventilated, hygienic, and protected pucca houses, with sufficient floor space. The cows were let out after morning milking (from 4:30 to 8:30 hrs), during which fresh drinking water was available. Additionally, fresh water was offered three times at the tying place (14:00–15:00, 17:00–18:00, and 21:00–22:00 hrs). All the crossbred cows were milked twice daily at 4:30 and 16:30 hours, using a pipeline milking machine. Body weight was measured weekly prior to feeding and watering on electronic weighbridge, while body condition score (BCS) was assessed four times: at 0 (Initial), 28, 56, and 70 day (Final), using a 1–5 point scale (Edmonson et al., 1989 ). The concentration of aflatoxin B1 (AFB1) in the concentrate mixture was quantified periodically at National Dairy Development Board, Centre for Analysis and Learning in Livestock and Food (NDDB CALF) laboratory, Anand using HPLC-Fluorescence. The 4% fat corrected milk (FCM) was calculated using the formula specified by Gains ( 1928 ) and energy-corrected milk (ECM) adjusted to 3.5% fat and 3.2% protein yield was calculated as per Tyrrell & Reid ( 1965 ) formula. Milk samples (100–150 ml) were collected weekly from individual cows during morning and evening milking, to measure milk fat, solid not fat (SNF), and protein using IndiFOSS Milkoscreen™ (BIS, 1981 ). Total solids were calculated as fat + SNF, and metabolizable energy (ME) of milk calculated using NRC ( 2001 ): ME (Mcal/kg) = 0.094×Fat% + 0.057×Protein% + 0.04×Lactose% The milk samples were collected from individual cows of all groups on 0 (Initial), 14, 28, and 70 (final) day of experiment and treatment group wise pooled milk samples were also collected on alternate days up to day 28, then on 42, 56, and 70 days to evaluate AFM1 levels. The AFM1 level was measured at NDDB CALF laboratory, Anand, using the standard method with HPLC-Fluorescence. The transfer of AFB1 into milk AFM1 was described as Bioconcentration, and carryover percentage were calculated as per Guo et al. ( 2021 ) and Patterson et al. ( 1980 ), respectively. The blood sample were aseptically drawn from the jugular vein of each crossbred cow into clot activator Vacuette in the morning before feeding and watering on 0, 28, 56, and 70 days of the study. Serum was separated by centrifugation at 550g for 15 minutes with a REMI research centrifuge and stored in sterilized Eppendorf tubes at -20°C for further testing. The separated serum was analysed for total protein (g/dl), albumin (g/dl), glucose (mg/dl), serum aspartate aminotransferase (AST)/ serum glutamic oxaloacetic transaminase (SGOT) (U/l), serum alanine aminotransferase (ALT)/ serum glutamic pyruvic transaminase (SGPT) (U/l), serum creatinine (mg/dl), and blood urea nitrogen (BUN) (mg/dl). The globulin (g/dl) and albumin to globulin ratio were calculated from the total protein and albumin values. All measurements were performed using the Merilyzer AutoQuant 200i VET machine, manufactured by Meril Diagnostics Private Limited, Gujarat, India. The data are presented as mean ± SE and analysed as per Snedecor & Cochran ( 2014 ) method as one-way ANOVA using SPSS 27. At probability (P) < 0.05, the difference between the means was considered significant. RESULTS Body Weight and Body Condition Score (BCS) Table 1 Body weight and body condition score of crossbred lactating cows Parameter T1 (Control) T2 (50 g/day/cow toxin binder) T3 (100 g/day/cow toxin binder) p -Value Initial body weight (kg) 469.97 ± 22.28 482.93 ± 14.65 510.02 ± 21.55 - Final body weight (kg) 467.73 ± 21.59 492.57 ± 18.44 501.07 ± 31.26 - Change in body weight (kg) -2.24 9.64 -8.95 - Body condition score 2.92 ± 0.06 2.97 ± 0.05 2.99 ± 0.04 0.582 The data on body weight, and BCS of lactating cows are presented in Table 1 , suggested the positive body weight change in T2 having 50 g/day toxin binder feeding. The BCS of cows of toxin binder group was also maintained. Milk and Components Yield Table 2 Milk and components yield in crossbred cows fed TMR with different levels of toxin binder Parameter T1 (Control) T2 (50 g/day/cow toxin binder) T3 (100 g/day/cow toxin binder) p -Value Milk yield (kg/day) 16.94 a ± 0.37 19.52 b ± 0.16 19.68 b ± 0.25 < 0.001 4% Fat corrected milk yield (kg/day) 17.89 a ± 0.35 20.53 c ± 0.24 19.34 b ± 0.34 < 0.001 Energy corrected milk yield (kg/day) 19.05 a ± 0.35 21.92 c ± 0.22 20.64 b ± 0.32 < 0.001 Milk fat (%) 4.45 b ± 0.11 4.35 b ± 0.07 3.91 a ± 0.11 < 0.001 Milk solids not fat (%) 8.59 b ± 0.06 8.55 b ± 0.03 8.24 a ± 0.03 < 0.001 Milk protein (%) 3.05 b ± 0.03 3.06 b ± 0.02 2.87 a ± 0.02 < 0.001 Milk total solids (%) 13.04 b ± 0.13 12.90 b ± 0.07 12.15 a ± 0.11 < 0.001 Milk fat yield (kg/day) 0.74 a ± 0.02 0.85 b ± 0.01 0.76 a ± 0.02 < 0.001 Milk solids not fat yield (kg/day) 1.45 a ± 0.03 1.67 b ± 0.02 1.62 b ± 0.02 < 0.001 Milk protein yield (g/day) 511.69 a ± 9.37 595.85 c ± 4.60 562.89 b ± 6.49 < 0.001 Milk total solids yield (kg/day) 2.19 a ± 0.04 2.52 c ± 0.02 2.38 b ± 0.03 < 0.001 Milk energy (Mcal/kg) 0.79 b ± 0.01 0.78 b ± 0.01 0.73 a ± 0.01 < 0.001 Milk energy yield (Mcal/day) 13.27 a ± 0.25 15.27 c ± 0.16 14.33 b ± 0.23 < 0.001 Where Mcal= Mega calorie Mean with different superscripts within row differed significantly (P < 0.05) The daily yield of milk, and milk components by crossbred cows are presented in Table 2 . The yield of milk, fat-corrected milk (FCM), and energy-corrected milk (ECM) were increased ( p < 0.001) in toxin binder-supplemented groups, with T2 consistently showing the highest improvements. Milk contents (% fat, protein, SNF, TS, and energy) were maintained in T2 but significantly reduced in T3 compared to control. The yields of milk fat and energy (Mcal/kg milk) improved ( p < 0.001) in T2, SNF in both T2 and T3 compared to control while yield of milk protein, total solids and milk energy (Mcal/day) were also significant higher in T2, T3 and highest value in T2. Aflatoxin B1 Intake and Aflatoxin M1 Excretion Table 3 Aflatoxin B1 intake and Aflatoxin M1 excretion in lactating crossbred cows fed TMR with different levels of toxin binder Parameter T1 (Control) T2 (50 g/day/cow toxin binder) T3 (100 g/day/cow toxin binder) p -Value Aflatoxin B1 in feed (µg/kg) 116.862 116.862 116.862 - Total aflatoxin B1 intake (µg/day) 2189.64 a ± 36.39 2466.89 b ± 28.24 (+ 12.66%) 2537.21 b ± 62.20 (+ 15.87%) < 0.001 Aflatoxin M1 Concentration in milk (µg/kg) 1.408 b ± 0.061 0.326 a ± 0.022 (-76.84%) 0.300 a ± 0.016 (-78.69%) < 0.001 Total aflatoxin M1 excretion in milk (µg/day/cow) 23.613 b ± 0.811 6.332 a ± 0.177 (-73.18%) 5.856 a ± 0.137 (-75.20%) < 0.001 Bio-concentration 0.012 b ± 0.001 0.003 a ± 0.000 (-75.00%) 0.003 a ± 0.000 (-75.00%) < 0.001 Carry-over % 1.077 b ± 0.032 0.257 a ± 0.008 (-76.14%) 0.234 a ± 0.008 (-78.27%) < 0.001 The average aflatoxin B1 in total mixed ration of lactating cows of all treatment groups was 116.862 µg/kg. Total AFB1 intake was higher ( p < 0.001) in T2 (12.66%) and T3 (15.87%) compared to T1 due to increased feed intake. Despite higher intake, toxin binder feeding markedly reduced AFM1 concentration (µg/kg) in milk of T2 and T3 groups (76.84% and 78.69%, respectively). Similarly, AFM1 excretion (µg/day) in milk was also lowered (P < 0.001) by 73.18% and 75.20% in T2 and T3, respectively compared to T1. The bio-concentration factor and carry-over percentage were also significantly reduced up to 78% with toxin binder feeding (Table 3 ). The concentration of aflatoxin M1 in milk (Fig. 1 ) started the decline from the sixth day and it was sharp after 10 days of supplemental feeding of toxin binder. The decline of AFM1 excretion in milk at the terminal part of study (70 days) was 76.66% and 75.66% (1.337 to 0.312 µg/kg milk and 1.426 to 0.347 µg/kg milk) in 50 (T2) and 100 g/day/cow (T3) toxin binder feeding groups, respectively compared to initial excretion before supplementation (0 day). Metabolic Health Table 4 Serum biochemical parameters of lactating crossbred cows fed TMR with different levels of toxin binder Parameter T1 (Control) T2 (50 g/day/cow toxin binder) T3 (100 g/day/cow toxin binder) P-Value Energy and protein nutrition metabolites Glucose (mg/dl) 56.70 ± 1.73 56.77 ± 1.20 55.67 ± 1.08 0.816 Total Protein (g/dl) 6.44 ± 0.14 6.68 ± 0.11 6.55 ± 0.09 0.318 Albumin (g/dl) 3.67 ± 0.07 3.82 ± 0.04 3.74 ± 0.05 0.165 Globulin (g/dl) 2.77 ± 0.14 2.86 ± 0.09 2.81 ± 0.06 0.822 A: G Ratio 1.40 ± 0.07 1.37 ± 0.04 1.34 ± 0.03 0.678 Liver function metabolites SGOT (U/l) 81.71 ± 2.81 81.91 ± 2.63 82.24 ± 2.49 0.990 SGPT (U/l) 30.39 ± 2.15 32.4 b ± 2.09 30.52 a ± 2.12 0.755 Kidney function metabolites Creatinine (mg/dl) 1.14 ± 0.03 1.11 ± 0.02 1.16 ± 0.03 0.322 BUN (mg/dl) 20.92 ± 0.61 19.92 ± 0.63 20.69 ± 0.63 0.498 Where, A:G = Albumin to globulin ratio; SGOT= Serum Glutamic Oxaloacetic Transaminase; SGPT= Serum Glutamic Pyruvic Transaminase; BUN= Blood Urea Nitrogen The serum biochemical metabolites including glucose, protein fractions (albumin & globulin), liver enzymes (SGOT, SGPT), and kidney function indicators (creatinine & BUN) of crossbred lactating cows (Table 4 ) were statistically not affected (P > 0.05) by toxin binder addition, and all values remained within normal physiological ranges (Hussein et al., 2020 ; Aiello & Moses, 2016 ). Treatment Cost Table 5 Treatment cost of lactating crossbred cows fed TMR with different levels of toxin binder Parameter T1 (Control) T2 (50 g/day/cow toxin binder) T3 (100 g/day/cow toxin binder) p -Value Treatment cost (₹/kg milk) 1.086 ± 0.029 0.030 ± 0.000 0.163 ± 0.002 < 0.001 Change over control - -1.056 (97.24%) -0.923 (-84.99%) - The treatment cost per unit of milk for different groups of lactating crossbred cows fed TMR supplemented with different levels of toxin binder is presented in Table 5 , showing a reduction ( p < 0.001) in treatment cost per unit of milk with toxin binder supplemented (T2 and T3) groups compared to control. DISCUSSION Body weight and body condition score (BCS) The toxin binder feeding at 50 g/day might have mitigated the adverse effects of dietary aflatoxins, resulted in improved intake and efficient nutrient utilization that were translated to higher milk synthesis and body weight gain. In contrast, the weight loss in T3, despite a higher initial body weight, may be due to reduced feed intake, utilization efficiency, metabolic adjustments, or palatability issues associated with a higher toxin binder dosage. Body condition scores remained unaffected across treatments, suggesting that moderate supplementation (T2) supported better energy balance. However, other studies have reported neutral or negative outcomes of toxin binder on body weight and BCS in lactating cows under a mycotoxin challenge (Lunagariya et al., 2024 ; Vieira et al., 2024 ; Maki et al., 2016 ). Milk and Components Yield The 50 g/day toxin binder feeding (T2) was more effective in improving yield of both milk and components. This might be the protective effect of toxin binder on liver that has improved intake and supply of nutrients for milk synthesis. Similar to these findings, Hajmohammadi et al. ( 2021 ) and Mehany & Shams ( 2019 ) observed improved ( p < 0.05) milk and components yield in Holstein and Friesian cows, respectively. However, Lunagariya et al. ( 2024 ), Vieira et al. ( 2024 ), Sherasia et al. ( 2023 ), and Guo et al. ( 2019 ) observed non-significant ( p > 0.05) improvement in milk and components yield on feeding diet supplemented with various toxin binders to lactating dairy cows. Aflatoxin B1 Intake and Aflatoxin M1 Excretion The lowered excretion of aflatoxin M1 in milk of toxin binder group (T2, T3) was attributed to the combined effects of HSCAS, bentonite, MOS, Bacillus DFM, and herbal components that restricted AFB1 absorption and metabolism to AFM1. Similarly, reduction in AFM1 concentration in milk was reported on feeding HSCAS, sodium bentonite, Bacillus subtilis , and calcium montmorillonite based toxin binder (Vieira et al., 2024 ; Sherasia et al., 2023 ; Hajmohammadi et al., 2021 ; Guo et al., 2019 ; Maki et al., 2016 ; Kutz et al., 2009 ) to dairy cows. While Xiong et al. ( 2015 ) noted only marginal decreases in concentration under high dietary AFB1, may be due to variable efficacy of toxin binder and contamination level. Both levels of supplementation were equally effective in reducing milk AFM1 below the maximum tolerance limit of 0.50 µg/kg (FSSAI, 2011 ; USFDA, 2018 ; Oliveira et al., 2014 ). The longer period (10 days) taken to reduce AFM1 excretion was due to the adaptation period taken by cows to completely clean up the toxin binder containing compound concentrate mixture. The supplemental feeding of 50 g/day/cow toxin binder was effective in reducing AFM1 excretion in milk and health risk to consumers. Metabolic Health The metabolites related to energy-protein nutrition, liver and kidney functions were within normal range and without significant effect of toxin binder, indicating safety of feeding and protecting effect on vital parameters physiological function on body. Similarly, non-significant (P > 0.05) changes in serum glucose, total protein, albumin, globulin, SGOT, SGPT, creatinine, BUN concentration reported on toxin binder supplementation (Vieira et al., 2024 ; Gallo et al., 2020 ; Guo et al., 2019 ) in dairy animals. Treatment Cost The reduced treatment cost per unit of milk in T2 group, highlight that adding 50 g/day toxin binder was most significant to accrue health benefit, and lowering treatment costs (97.24%). CONCLUSION Cows fed total mixed ration and 50 g/day toxin binder (T2) showed positive body weight gain, higher yield of milk, fat-corrected milk, energy-corrected milk, and milk components with maintained milk components level compared to control. Despite greater aflatoxin B1 intake from higher feed consumption, supplementation reduced concentration, daily excretion, bioconcentration, and carry-over of aflatoxin M1in milk by 76.84, 73.18, 75.00, and 76.14%, respectively over control. The toxin binder also not influenced liver, kidney function and energy-protein nutrition. The treatment cost per unit milk production was also reduced by 97.24%, indicating health benefits. The toxin binder at higher dose rate (100 g/day) was without further benefits on measured parameters. Thus, feeding of 50 g/day toxin binder proved effective in enhancing performance and welfare of cow while minimizing aflatoxin health risk to consumers. Declarations Acknowledgment We gratefully acknowledge College of Veterinary Science and Animal Husbandry, Kamdhenu University, Kamdhenu University, Anand, Gujarat, India for providing exceptional support and research facilities that were instrumental in enabling the successful completion of this work. Author contributions : PML, BRD, JHP: formulated research; supervise research, HA: executed research, laboratory analysis and data analysis, AMS, HA: Helped in laboratory analysis, wrote manuscript, JHP, DGV: wrote & revised manuscript. All authors approved the manuscript. Ethical approval: An experiment protocol was approved by Institutional Animal Ethics Committee (IAEC) of College of Veterinary Science & Animal Husbandry, Kamdhenu University, Anand (Gujarat, India) and cows were cared accordingly. The experiment was conducted at Livestock Research Station, College of Veterinary Science & Animal Husbandry, Kamdhenu University, Anand, Gujarat, India. Conflict of Interest: This was contractual research and funded by JP Veterinary, Mehsana, Gujarat, India. Data availability : The datasets of the current study will available from the corresponding author on reasonable request. References Aiello, S. E., and Moses, M.A. (2016). The Merck Veterinary Manual. 11 th ed., 3176-3181. 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Toxins 8(9): 245. https://doi.org/10.3390/toxins8090245 Lunagariya PM, Parmar CP, Trivedi PG, Desai DM, Agravat PH, Sorathiya KK, Wadhwani KN (2024) Ameliorating effects of Mycotoxcease (Mycotoxin binder) on aflatoxin M1 excretion in dairy cows. Indian Journal of Veterinary Science and Biotechnology 20(6): 114–119. https://doi.org/10.48165/ijvsbt.20.6.22 Maki CR, Thomas AD, Elmore SE, Romoser AA, Harvey RB, Ramirez-Ramirez HA, Phillips TD (2016) Effects of calcium montmorillonite clay and aflatoxin exposure on dry matter intake, milk production, and milk composition. Journal of Dairy Science 99(2): 1039–1046. https://doi.org/10.3168/jds.2015-9988 Mehany A, Shams A (2019) Effect of toxin binder on productive performance of lactating Friesian cows. Journal of Animal and Poultry Production 10(12): 405–413. DOI:10.21608/jappmu.2019.82461 NRC (2001). Nutrient requirements of dairy cattle (7 th ed.). National Academies Press, National Research Council, Washington, DC, USA Oliveira CAF, Corassin CH, Oswald C (2014) Animal health: mycotoxins. In K. Neal & V. Alfen (Eds.), Encyclopedia of Agriculture and Food Systems Pg. 358–377. Elsevier Limited. Patterson DS, Glancy EM, Roberts BA (1980) The 'carry over' of aflatoxin M1 into the milk of cows fed rations containing a low concentration of aflatoxin B1. Food and cosmetics toxicology 18(1): 35–37. 10.1016/0015-6264(80)90008-5 Queiroz OCM, Han JH, Staples CR, Adesogan AT (2012) Effect of adding a mycotoxin-sequestering agent on milk aflatoxin M1 concentration and the performance and immune response of dairy cattle fed an aflatoxin B1-contaminated diet. Journal of Dairy Science 95(10): 5901–5908. https://doi.org/10.3168/jds.2011-5287 Sherasia P L, Waghela CR, Sharma R, Patel AP, Sridhar V, Pawar PM, Gupta SK (2023) Effect of feeding bentonite clay on excretion of aflatoxin M1 in milk of dairy animals. Indian Journal of Animal Nutrition 40(3): 296–304. https://doi.org/10.5958/2231-6744.2023.00037.3 Snedecor GW, Cochran WG (2014) Statistical methods (8 th Ed.). Oxford and IBH Publishing Co., Calcutta, India. DOI: https://doi.org/10.1017/S0021859600074104 Tolosa J, Rodríguez-Carrasco Y, Ruiz MJ, Vila-Donat P (2021) Multi-mycotoxin occurrence in feed, metabolism and carry over to animal-derived food products: A review. Food and Chemical Toxicology 158: 112661. https://doi.org/10.1016/j.fct.2021.112661 Tyrrell HF, Reid JT (1965) Prediction of the energy value of cow’s milk. Journal of Dairy Science 48: 1215–1223. https://doi.org/10.3168/jds.S0022-0302(65)88384-9 USFDA (2018) United States Food and Drug Administration, United State of America. Retrieved from https://www.fda.gov/AnimalVeterinary/Products/ucm050223. htm. Veldman A, Meijs JAC, Borggreve GJ, Tol JJHD (1992) Carry-over of aflatoxin from cows’ food to milk. Animal Science 55(2): 163–168. https://doi.org/10.1017/S0003356100037417 Vieira DJC, Fonseca LM, Poletti G, Martins NP, Grigoletto NTS, Chesini RG, Tonin FG, Cortinhas CS, Acedo TS, Artavia I, Faas J, Takiya CS, Corassin CH, Rennó FP (2024) Anti-mycotoxin feed additives: Effects on metabolism, mycotoxin excretion, performance, and total-tract digestibility of dairy cows fed artificially multi-mycotoxin-contaminated diets. Journal of Dairy Science 107(10): 7891–7903. https://doi.org/10.3168/jds.2023-24539 Xiong JL, Wang YM, Nennich TD, Li Y, Liu JX (2015) Transfer of dietary aflatoxin B1 to milk aflatoxin M1 and effect of inclusion of adsorbent in the diet of dairy cows. Journal of Dairy Science 98(4): 2545–2554. https://doi.org/10.3168/jds.2013-7842 Additional Declarations Competing interest reported. This was contractual research. The product and fund provided by by JP Veterinary, Mehsana, Gujarat, India. Cite Share Download PDF Status: Under Review Version 1 posted Reviews received at journal 24 Apr, 2026 Reviewers agreed at journal 13 Apr, 2026 Reviewers invited by journal 08 Apr, 2026 Editor assigned by journal 16 Mar, 2026 Submission checks completed at journal 16 Mar, 2026 First submitted to journal 05 Mar, 2026 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-9039431\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":false,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":623624924,\"identity\":\"68f43abc-10e8-4bc1-92ae-b3bea10566ad\",\"order_by\":0,\"name\":\"Himansu Agrawal\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Kamdhenu University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Himansu\",\"middleName\":\"\",\"lastName\":\"Agrawal\",\"suffix\":\"\"},{\"id\":623624930,\"identity\":\"f6a46687-a6fb-4e62-ad7a-033a5c922655\",\"order_by\":1,\"name\":\"Bharat R. Devalia\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Kamdhenu University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Bharat\",\"middleName\":\"R.\",\"lastName\":\"Devalia\",\"suffix\":\"\"},{\"id\":623624934,\"identity\":\"a4e2aee5-e43c-4bb0-bcb8-07f165970b03\",\"order_by\":2,\"name\":\"Akash M. Sutariya\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Kamdhenu University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Akash\",\"middleName\":\"M.\",\"lastName\":\"Sutariya\",\"suffix\":\"\"},{\"id\":623624938,\"identity\":\"5942ad42-ddf4-41eb-9545-2266d6de6bbc\",\"order_by\":3,\"name\":\"Jigar H. Patel\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Kamdhenu University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Jigar\",\"middleName\":\"H.\",\"lastName\":\"Patel\",\"suffix\":\"\"},{\"id\":623624939,\"identity\":\"14e52b95-889c-445f-854e-d436b76b3f1c\",\"order_by\":4,\"name\":\"Dilip G. 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Lunagariya\",\"email\":\"data:image/png;base64,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\",\"orcid\":\"\",\"institution\":\"Kamdhenu University\",\"correspondingAuthor\":true,\"prefix\":\"\",\"firstName\":\"Pravin\",\"middleName\":\"M.\",\"lastName\":\"Lunagariya\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2026-03-05 11:23:31\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-9039431/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-9039431/v1\",\"draftVersion\":[],\"editorialEvents\":[],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":107088691,\"identity\":\"8ca550ba-d963-40b3-bba4-04d65b803a65\",\"added_by\":\"auto\",\"created_at\":\"2026-04-16 15:30:11\",\"extension\":\"png\",\"order_by\":1,\"title\":\"Figure 1\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":53764,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e\\u003cstrong\\u003eAFM1 concentration (µg/kg) in milk of crossbred lactating cows\\u003c/strong\\u003e\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"1.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-9039431/v1/a4e303439f78fe44c0bf3c84.png\"},{\"id\":107704940,\"identity\":\"f0e589d7-68cc-478d-98e8-0f4f7a01a78e\",\"added_by\":\"auto\",\"created_at\":\"2026-04-24 09:04:39\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":433413,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-9039431/v1/89cacd46-fb95-4e43-bf95-7cc876f59f15.pdf\"}],\"financialInterests\":\"Competing interest reported. This was contractual research. The product and fund provided by by JP Veterinary, Mehsana, Gujarat, India.\",\"formattedTitle\":\"Effect of toxin binder feeding on body weight, milk and components yield, and aflatoxin M1 excretion in crossbred lactating cows\",\"fulltext\":[{\"header\":\"INTRODUCTION\",\"content\":\"\\u003cp\\u003eMycotoxin contamination of livestock feed poses a significant threat to animal health, productivity, and food safety. Dairy animals consume a wide variety of feedstuffs, including concentrates, silages, and forages, which are vulnerable to fungal contamination. Aflatoxins, produced by \\u003cem\\u003eAspergillus flavus, A. parasiticus\\u003c/em\\u003e, and \\u003cem\\u003eA. nomius\\u003c/em\\u003e, are the most harmful mycotoxins (Gon\\u0026ccedil;alves et al., \\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e2017\\u003c/span\\u003e). Aflatoxin B1 (AFB1) is the most potent, classified by the International Agency for Research on Cancer as a Group I human carcinogen (IARC, \\u003cspan citationid=\\\"CR19\\\" class=\\\"CitationRef\\\"\\u003e2002\\u003c/span\\u003e). In ruminants, AFB1 is partly degraded in the rumen (Tolosa et al., \\u003cspan citationid=\\\"CR31\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e), but the absorbed fraction undergoes hepatic metabolism, generating toxic intermediates that damage DNA, proteins, and lipids (Cupid et al., \\u003cspan citationid=\\\"CR6\\\" class=\\\"CitationRef\\\"\\u003e2004\\u003c/span\\u003e). In dairy cows, AFB1 is rapidly bio-transformed into aflatoxin M1 (AFM1), which appears in milk within hours of ingestion (Guo et al., \\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e; Gallo et al., \\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e2008\\u003c/span\\u003e; Battacone et al., \\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2003\\u003c/span\\u003e). Carry-over rates range from 1% to 6%, depending on milk yield, breed, and stage of lactation (Campagnollo et al., \\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e; Battacone et al., \\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2003\\u003c/span\\u003e; Veldman et al., \\u003cspan citationid=\\\"CR34\\\" class=\\\"CitationRef\\\"\\u003e1992\\u003c/span\\u003e). The impact of mycotoxin on hematological and biochemical indices is also limited.\\u003c/p\\u003e \\u003cp\\u003eSeveral strategies have been explored to mitigate the excretion of aflatoxin in milk. Among them, toxin binder supplementation remains the effective and practical on-farm approach. Adsorbents such as bentonite, montmorillonite, hydrated sodium calcium aluminosilicate (HSCAS), zeolites, activated charcoal, and yeast cell wall components are widely used (Čolović et al., \\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e). Among these, HSCAS has demonstrated significant effectiveness, binding up to 95% of aflatoxins and nearly halving AFM1 excretion in milk (Queiroz et al., \\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e2012\\u003c/span\\u003e; Kutz et al., \\u003cspan citationid=\\\"CR20\\\" class=\\\"CitationRef\\\"\\u003e2009\\u003c/span\\u003e). At the same time, biological detoxification using microorganisms such as \\u003cem\\u003eBacillus subtilis, Rhodococcus erythropolis\\u003c/em\\u003e, and \\u003cem\\u003ePleurotus pulmonarius\\u003c/em\\u003e has shown promise in degrading aflatoxins, although further \\u003cem\\u003ein vivo\\u003c/em\\u003e validation is required (Loi et al., \\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e; Eshelli et al., \\u003cspan citationid=\\\"CR8\\\" class=\\\"CitationRef\\\"\\u003e2015\\u003c/span\\u003e; Gao et al., \\u003cspan citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e2011\\u003c/span\\u003e). The toxin binder evaluated in this study consisted of bentonite, mannan oligosaccharides (MOS), \\u003cem\\u003eBacillus\\u003c/em\\u003e (direct-fed microbial), organic acids, oxine copper, and herbal extracts, designed to provide complementary benefits, including toxin adsorption, microbial detoxification, immune modulation, gut environment stabilization, and hepato-protective effects. The study was conducted to evaluate the effects of toxin binder supplementation on body weight, body condition score, milk and components yield as well as on aflatoxin excretion in milk of lactating crossbred cows.\\u003c/p\\u003e\"},{\"header\":\"MATERIALS AND METHODS\",\"content\":\"\\u003cp\\u003eThe experiment was conducted for 70 days, following a ten-day adaptive feeding period. A total of eighteen 75% Holstein Friesian x 25% Kankrej- Indian Zebu crossbred lactating cows were randomly allotted to 3 treatment groups (6 in each) based on similarity of milk yield (19.09\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.21 kg/day), days in milk (71.8\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;14.3 days), and lactation numbers (2.11\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.19). The cows of three treatment groups \\u003cem\\u003eviz\\u003c/em\\u003e. Control (T1), Treatment 2 (T2), and Treatment 3 (T3) were fed \\u003cem\\u003ead-libitum\\u003c/em\\u003e total mixed ration (TMR). The composite toxin binder containing hydrated sodium calcium alumino-silicate (bentonite), organic acids, mannan oligosaccharides, \\u003cem\\u003eBacillus\\u003c/em\\u003e direct-fed microbial, oxine copper, and herbal ingredients fed to cows of T2 and T3 groups at 50 g/day and 100 g/day, respectively. The toxin binder fed to cows in equal half after mixing with 250 g compounded concentrate mixture (CCM) in morning and evening in treatment groups and equal quantity of CCM also fed to cows of control group. The CCM kept open before feeding, AFM1 excretion in milk before and after feeding were measured for this experiment. The TMR was prepared using 42kg compounded concentrate mixture, 13kg wheat straw, 24kg groundnut straw, 15kg hybrid Napier, 4kg bypass fat, 1kg mineral-vitamin premix, and 1kg salt. All the crossbred cows were housed in well-ventilated, hygienic, and protected pucca houses, with sufficient floor space. The cows were let out after morning milking (from 4:30 to 8:30 hrs), during which fresh drinking water was available. Additionally, fresh water was offered three times at the tying place (14:00\\u0026ndash;15:00, 17:00\\u0026ndash;18:00, and 21:00\\u0026ndash;22:00 hrs). All the crossbred cows were milked twice daily at 4:30 and 16:30 hours, using a pipeline milking machine.\\u003c/p\\u003e \\u003cp\\u003eBody weight was measured weekly prior to feeding and watering on electronic weighbridge, while body condition score (BCS) was assessed four times: at 0 (Initial), 28, 56, and 70 day (Final), using a 1\\u0026ndash;5 point scale (Edmonson et al., \\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e1989\\u003c/span\\u003e). The concentration of aflatoxin B1 (AFB1) in the concentrate mixture was quantified periodically at National Dairy Development Board, Centre for Analysis and Learning in Livestock and Food (NDDB CALF) laboratory, Anand using HPLC-Fluorescence. The 4% fat corrected milk (FCM) was calculated using the formula specified by Gains (\\u003cspan citationid=\\\"CR10\\\" class=\\\"CitationRef\\\"\\u003e1928\\u003c/span\\u003e) and energy-corrected milk (ECM) adjusted to 3.5% fat and 3.2% protein yield was calculated as per Tyrrell \\u0026amp; Reid (\\u003cspan citationid=\\\"CR32\\\" class=\\\"CitationRef\\\"\\u003e1965\\u003c/span\\u003e) formula. Milk samples (100\\u0026ndash;150 ml) were collected weekly from individual cows during morning and evening milking, to measure milk fat, solid not fat (SNF), and protein using IndiFOSS Milkoscreen\\u0026trade; (BIS, \\u003cspan citationid=\\\"CR3\\\" class=\\\"CitationRef\\\"\\u003e1981\\u003c/span\\u003e). Total solids were calculated as fat\\u0026thinsp;+\\u0026thinsp;SNF, and metabolizable energy (ME) of milk calculated using NRC (\\u003cspan citationid=\\\"CR25\\\" class=\\\"CitationRef\\\"\\u003e2001\\u003c/span\\u003e):\\u003c/p\\u003e \\u003cp\\u003eME (Mcal/kg)\\u0026thinsp;=\\u0026thinsp;0.094\\u0026times;Fat% + 0.057\\u0026times;Protein% + 0.04\\u0026times;Lactose%\\u003c/p\\u003e \\u003cp\\u003eThe milk samples were collected from individual cows of all groups on 0 (Initial), 14, 28, and 70 (final) day of experiment and treatment group wise pooled milk samples were also collected on alternate days up to day 28, then on 42, 56, and 70 days to evaluate AFM1 levels. The AFM1 level was measured at NDDB CALF laboratory, Anand, using the standard method with HPLC-Fluorescence. The transfer of AFB1 into milk AFM1 was described as Bioconcentration, and carryover percentage were calculated as per Guo et al. (\\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e) and Patterson et al. (\\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e1980\\u003c/span\\u003e), respectively.\\u003c/p\\u003e \\u003cp\\u003eThe blood sample were aseptically drawn from the jugular vein of each crossbred cow into clot activator Vacuette in the morning before feeding and watering on 0, 28, 56, and 70 days of the study. Serum was separated by centrifugation at 550g for 15 minutes with a REMI research centrifuge and stored in sterilized Eppendorf tubes at -20\\u0026deg;C for further testing. The separated serum was analysed for total protein (g/dl), albumin (g/dl), glucose (mg/dl), serum aspartate aminotransferase (AST)/ serum glutamic oxaloacetic transaminase (SGOT) (U/l), serum alanine aminotransferase (ALT)/ serum glutamic pyruvic transaminase (SGPT) (U/l), serum creatinine (mg/dl), and blood urea nitrogen (BUN) (mg/dl). The globulin (g/dl) and albumin to globulin ratio were calculated from the total protein and albumin values. All measurements were performed using the Merilyzer AutoQuant 200i VET machine, manufactured by Meril Diagnostics Private Limited, Gujarat, India.\\u003c/p\\u003e \\u003cp\\u003eThe data are presented as mean\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;SE and analysed as per Snedecor \\u0026amp; Cochran (\\u003cspan citationid=\\\"CR30\\\" class=\\\"CitationRef\\\"\\u003e2014\\u003c/span\\u003e) method as one-way ANOVA using SPSS 27. At probability (P)\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05, the difference between the means was considered significant.\\u003c/p\\u003e\"},{\"header\":\"RESULTS\",\"content\":\"\\u003cdiv id=\\\"Sec4\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eBody Weight and Body Condition Score (BCS)\\u003c/h2\\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\\u003eBody weight and body condition score of crossbred lactating cows\\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\\u003eParameter\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eT1 (Control)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eT2 (50 g/day/cow toxin binder)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eT3 (100 g/day/cow toxin binder)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003ep\\u003c/em\\u003e-Value\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eInitial body weight (kg)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e469.97\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;22.28\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e482.93\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;14.65\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e510.02\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;21.55\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e-\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eFinal body weight (kg)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e467.73\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;21.59\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e492.57\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;18.44\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e501.07\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;31.26\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e-\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eChange in body weight (kg)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e-2.24\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.64\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e-8.95\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e-\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eBody condition score\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e2.92\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.06\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e2.97\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.05\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e2.99\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.04\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.582\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003eThe data on body weight, and BCS of lactating cows are presented in Table\\u0026nbsp;\\u003cspan refid=\\\"Tab1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e, suggested the positive body weight change in T2 having 50 g/day toxin binder feeding. The BCS of cows of toxin binder group was also maintained.\\u003c/p\\u003e \\u003c/div\\u003e\\n\\u003ch3\\u003eMilk and Components Yield\\u003c/h3\\u003e\\n\\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\\u003eMilk and components yield in crossbred cows fed TMR with different levels of toxin binder\\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=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eParameter\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eT1 (Control)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eT2 (50 g/day/cow toxin binder)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eT3 (100 g/day/cow toxin binder)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003ep\\u003c/em\\u003e-Value\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eMilk yield (kg/day)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e16.94\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.37\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e19.52\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.16\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e19.68\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003e4% Fat corrected milk yield (kg/day)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e17.89\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.35\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e20.53\\u003csup\\u003ec\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.24\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e19.34\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.34\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eEnergy corrected milk yield (kg/day)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e19.05\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.35\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e21.92\\u003csup\\u003ec\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.22\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e20.64\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.32\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eMilk fat (%)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e4.45\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.11\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e4.35\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.07\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e3.91\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.11\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eMilk solids not fat (%)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e8.59\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.06\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e8.55\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.03\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e8.24\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.03\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eMilk protein (%)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e3.05\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.03\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e3.06\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.02\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e2.87\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.02\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eMilk total solids (%)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e13.04\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.13\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e12.90\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.07\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e12.15\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.11\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eMilk fat yield (kg/day)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e0.74\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.02\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.85\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.76\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.02\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eMilk solids not fat yield (kg/day)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1.45\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.03\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e1.67\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.02\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e1.62\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.02\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eMilk protein yield (g/day)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e511.69\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e \\u003cp\\u003e\\u0026plusmn;\\u0026thinsp;9.37\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e595.85\\u003csup\\u003ec\\u003c/sup\\u003e\\u003c/p\\u003e \\u003cp\\u003e\\u0026plusmn;\\u0026thinsp;4.60\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e562.89\\u003csup\\u003eb\\u003c/sup\\u003e\\u003c/p\\u003e \\u003cp\\u003e\\u0026plusmn;\\u0026thinsp;6.49\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eMilk total solids yield (kg/day)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e2.19\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.04\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e2.52\\u003csup\\u003ec\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.02\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e2.38\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.03\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eMilk energy (Mcal/kg)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e0.79\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.78\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.73\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eMilk energy yield (Mcal/day)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e13.27\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e15.27\\u003csup\\u003ec\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.16\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e14.33\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.23\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003ctfoot\\u003e \\u003ctr\\u003e\\u003ctd colspan=\\\"5\\\"\\u003eWhere Mcal= Mega calorie\\u003c/td\\u003e\\u003c/tr\\u003e \\u003c/tfoot\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003eMean with different superscripts within row differed significantly (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05)\\u003c/p\\u003e \\u003cp\\u003eThe daily yield of milk, and milk components by crossbred cows are presented in Table\\u0026nbsp;\\u003cspan refid=\\\"Tab2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e. The yield of milk, fat-corrected milk (FCM), and energy-corrected milk (ECM) were increased (\\u003cem\\u003ep\\u003c/em\\u003e\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.001) in toxin binder-supplemented groups, with T2 consistently showing the highest improvements. Milk contents (% fat, protein, SNF, TS, and energy) were maintained in T2 but significantly reduced in T3 compared to control. The yields of milk fat and energy (Mcal/kg milk) improved (\\u003cem\\u003ep\\u003c/em\\u003e\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.001) in T2, SNF in both T2 and T3 compared to control while yield of milk protein, total solids and milk energy (Mcal/day) were also significant higher in T2, T3 and highest value in T2.\\u003c/p\\u003e\\n\\u003ch3\\u003eAflatoxin B1 Intake and Aflatoxin M1 Excretion\\u003c/h3\\u003e\\n\\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\\u003eAflatoxin B1 intake and Aflatoxin M1 excretion in lactating crossbred cows fed TMR with different levels of toxin binder\\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\\u003eParameter\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eT1 (Control)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eT2 (50 g/day/cow toxin binder)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eT3 (100 g/day/cow toxin binder)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003ep\\u003c/em\\u003e-Value\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eAflatoxin B1 in feed (\\u0026micro;g/kg)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e116.862\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e116.862\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e116.862\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e-\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eTotal aflatoxin B1 intake (\\u0026micro;g/day)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e2189.64\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;36.39\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e2466.89\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;28.24\\u003c/p\\u003e \\u003cp\\u003e(+\\u0026thinsp;12.66%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e2537.21\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;62.20\\u003c/p\\u003e \\u003cp\\u003e(+\\u0026thinsp;15.87%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eAflatoxin M1 Concentration in milk (\\u0026micro;g/kg)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1.408\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.061\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.326\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.022\\u003c/p\\u003e \\u003cp\\u003e(-76.84%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.300\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.016\\u003c/p\\u003e \\u003cp\\u003e(-78.69%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eTotal aflatoxin M1 excretion in milk (\\u0026micro;g/day/cow)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e23.613\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.811\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e6.332\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.177\\u003c/p\\u003e \\u003cp\\u003e(-73.18%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e5.856\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.137\\u003c/p\\u003e \\u003cp\\u003e(-75.20%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eBio-concentration\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e0.012\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.003\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.000\\u003c/p\\u003e \\u003cp\\u003e(-75.00%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.003\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.000\\u003c/p\\u003e \\u003cp\\u003e(-75.00%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eCarry-over %\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1.077\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.032\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.257 \\u003csup\\u003ea\\u003c/sup\\u003e \\u0026plusmn; 0.008\\u003c/p\\u003e \\u003cp\\u003e(-76.14%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.234 \\u003csup\\u003ea\\u003c/sup\\u003e \\u0026plusmn; 0.008\\u003c/p\\u003e \\u003cp\\u003e(-78.27%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003eThe average aflatoxin B1 in total mixed ration of lactating cows of all treatment groups was 116.862 \\u0026micro;g/kg. Total AFB1 intake was higher (\\u003cem\\u003ep\\u003c/em\\u003e\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.001) in T2 (12.66%) and T3 (15.87%) compared to T1 due to increased feed intake. Despite higher intake, toxin binder feeding markedly reduced AFM1 concentration (\\u0026micro;g/kg) in milk of T2 and T3 groups (76.84% and 78.69%, respectively). Similarly, AFM1 excretion (\\u0026micro;g/day) in milk was also lowered (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.001) by 73.18% and 75.20% in T2 and T3, respectively compared to T1. The bio-concentration factor and carry-over percentage were also significantly reduced up to 78% with toxin binder feeding (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003eThe concentration of aflatoxin M1 in milk (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e) started the decline from the sixth day and it was sharp after 10 days of supplemental feeding of toxin binder. The decline of AFM1 excretion in milk at the terminal part of study (70 days) was 76.66% and 75.66% (1.337 to 0.312 \\u0026micro;g/kg milk and 1.426 to 0.347 \\u0026micro;g/kg milk) in 50 (T2) and 100 g/day/cow (T3) toxin binder feeding groups, respectively compared to initial excretion before supplementation (0 day).\\u003c/p\\u003e\\n\\u003ch3\\u003eMetabolic Health\\u003c/h3\\u003e\\n\\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab4\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 4\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eSerum biochemical parameters of lactating crossbred cows fed TMR with different levels of toxin binder\\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\\u003eParameter\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eT1\\u003c/p\\u003e \\u003cp\\u003e(Control)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eT2\\u003c/p\\u003e \\u003cp\\u003e(50 g/day/cow toxin binder)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eT3\\u003c/p\\u003e \\u003cp\\u003e(100 g/day/cow toxin binder)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eP-Value\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colspan=\\\"5\\\" nameend=\\\"c5\\\" namest=\\\"c1\\\"\\u003e \\u003cp\\u003eEnergy and protein nutrition metabolites\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eGlucose (mg/dl)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e56.70\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.73\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e56.77\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.20\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e55.67\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.08\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.816\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eTotal Protein (g/dl)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e6.44\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.14\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e6.68\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.11\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e6.55\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.09\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.318\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eAlbumin (g/dl)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e3.67\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.07\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e3.82\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.04\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e3.74\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.05\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.165\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eGlobulin (g/dl)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e2.77\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.14\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e2.86\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.09\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e2.81\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.06\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.822\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eA: G\\u0026nbsp;Ratio\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1.40\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.07\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e1.37\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.04\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e1.34\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.03\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.678\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"5\\\" nameend=\\\"c5\\\" namest=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eLiver function metabolites\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eSGOT (U/l)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e81.71\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2.81\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e81.91\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2.63\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e82.24\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2.49\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.990\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eSGPT (U/l)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e30.39\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2.15\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e32.4\\u003csup\\u003eb\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2.09\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e30.52\\u003csup\\u003ea\\u003c/sup\\u003e\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2.12\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.755\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"5\\\" nameend=\\\"c5\\\" namest=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eKidney function metabolites\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eCreatinine (mg/dl)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1.14\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.03\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e1.11\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.02\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e1.16\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.03\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.322\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eBUN (mg/dl)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e20.92\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.61\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e19.92\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.63\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e20.69\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.63\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.498\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003ctfoot\\u003e \\u003ctr\\u003e\\u003ctd colspan=\\\"5\\\"\\u003eWhere, A:G\\u0026thinsp;=\\u0026thinsp;Albumin to globulin ratio; SGOT= Serum Glutamic Oxaloacetic Transaminase; SGPT= Serum Glutamic Pyruvic Transaminase; BUN= Blood Urea Nitrogen\\u003c/td\\u003e\\u003c/tr\\u003e \\u003c/tfoot\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"BlockQuote\\\"\\u003e \\u003cp\\u003eThe serum biochemical metabolites including glucose, protein fractions (albumin \\u0026amp; globulin), liver enzymes (SGOT, SGPT), and kidney function indicators (creatinine \\u0026amp; BUN) of crossbred lactating cows (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003e) were statistically not affected (P\\u0026thinsp;\\u0026gt;\\u0026thinsp;0.05) by toxin binder addition, and all values remained within normal physiological ranges (Hussein et al., \\u003cspan citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e; Aiello \\u0026amp; Moses, \\u003cspan citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e).\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/p\\u003e \\u003cdiv id=\\\"Sec8\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eTreatment Cost\\u003c/h2\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab5\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 5\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eTreatment cost of lactating crossbred cows fed TMR with different levels of toxin binder\\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\\u003eParameter\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eT1\\u003c/p\\u003e \\u003cp\\u003e(Control)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eT2\\u003c/p\\u003e \\u003cp\\u003e(50 g/day/cow toxin binder)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eT3\\u003c/p\\u003e \\u003cp\\u003e(100 g/day/cow toxin binder)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003ep\\u003c/em\\u003e-Value\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eTreatment cost (₹/kg milk)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1.086\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.029\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.030\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.000\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.163\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.002\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.001\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eChange over control\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e-\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-1.056\\u003c/p\\u003e \\u003cp\\u003e(97.24%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e-0.923\\u003c/p\\u003e \\u003cp\\u003e(-84.99%)\\u003c/p\\u003e\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e-\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003eThe treatment cost per unit of milk for different groups of lactating crossbred cows fed TMR supplemented with different levels of toxin binder is presented in Table\\u0026nbsp;\\u003cspan refid=\\\"Tab5\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003e, showing a reduction (\\u003cem\\u003ep\\u003c/em\\u003e\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.001) in treatment cost per unit of milk with toxin binder supplemented (T2 and T3) groups compared to control.\\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"DISCUSSION\",\"content\":\"\\u003cdiv id=\\\"Sec10\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eBody weight and body condition score (BCS)\\u003c/h2\\u003e \\u003cp\\u003eThe toxin binder feeding at 50 g/day might have mitigated the adverse effects of dietary aflatoxins, resulted in improved intake and efficient nutrient utilization that were translated to higher milk synthesis and body weight gain. In contrast, the weight loss in T3, despite a higher initial body weight, may be due to reduced feed intake, utilization efficiency, metabolic adjustments, or palatability issues associated with a higher toxin binder dosage. Body condition scores remained unaffected across treatments, suggesting that moderate supplementation (T2) supported better energy balance. However, other studies have reported neutral or negative outcomes of toxin binder on body weight and BCS in lactating cows under a mycotoxin challenge (Lunagariya et al., \\u003cspan citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e2024\\u003c/span\\u003e; Vieira et al., \\u003cspan citationid=\\\"CR35\\\" class=\\\"CitationRef\\\"\\u003e2024\\u003c/span\\u003e; Maki et al., \\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e).\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec11\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eMilk and Components Yield\\u003c/h2\\u003e \\u003cp\\u003eThe 50 g/day toxin binder feeding (T2) was more effective in improving yield of both milk and components. This might be the protective effect of toxin binder on liver that has improved intake and supply of nutrients for milk synthesis. Similar to these findings, Hajmohammadi et al. (\\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e) and Mehany \\u0026amp; Shams (\\u003cspan citationid=\\\"CR24\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e) observed improved (\\u003cem\\u003ep\\u003c/em\\u003e\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05) milk and components yield in Holstein and Friesian cows, respectively. However, Lunagariya et al. (\\u003cspan citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e2024\\u003c/span\\u003e), Vieira et al. (\\u003cspan citationid=\\\"CR35\\\" class=\\\"CitationRef\\\"\\u003e2024\\u003c/span\\u003e), Sherasia et al. (\\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e), and Guo et al. (\\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e) observed non-significant (\\u003cem\\u003ep\\u003c/em\\u003e\\u0026thinsp;\\u0026gt;\\u0026thinsp;0.05) improvement in milk and components yield on feeding diet supplemented with various toxin binders to lactating dairy cows.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec12\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eAflatoxin B1 Intake and Aflatoxin M1 Excretion\\u003c/h2\\u003e \\u003cp\\u003eThe lowered excretion of aflatoxin M1 in milk of toxin binder group (T2, T3) was attributed to the combined effects of HSCAS, bentonite, MOS, \\u003cem\\u003eBacillus\\u003c/em\\u003e DFM, and herbal components that restricted AFB1 absorption and metabolism to AFM1. Similarly, reduction in AFM1 concentration in milk was reported on feeding HSCAS, sodium bentonite, \\u003cem\\u003eBacillus subtilis\\u003c/em\\u003e, and calcium montmorillonite based toxin binder (Vieira et al., \\u003cspan citationid=\\\"CR35\\\" class=\\\"CitationRef\\\"\\u003e2024\\u003c/span\\u003e; Sherasia et al., \\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e; Hajmohammadi et al., \\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e; Guo et al., \\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e; Maki et al., \\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e; Kutz et al., \\u003cspan citationid=\\\"CR20\\\" class=\\\"CitationRef\\\"\\u003e2009\\u003c/span\\u003e) to dairy cows. While Xiong et al. (\\u003cspan citationid=\\\"CR36\\\" class=\\\"CitationRef\\\"\\u003e2015\\u003c/span\\u003e) noted only marginal decreases in concentration under high dietary AFB1, may be due to variable efficacy of toxin binder and contamination level. Both levels of supplementation were equally effective in reducing milk AFM1 below the maximum tolerance limit of 0.50 \\u0026micro;g/kg (FSSAI, \\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e2011\\u003c/span\\u003e; USFDA, \\u003cspan citationid=\\\"CR33\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e; Oliveira et al., \\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e2014\\u003c/span\\u003e). The longer period (10 days) taken to reduce AFM1 excretion was due to the adaptation period taken by cows to completely clean up the toxin binder containing compound concentrate mixture. The supplemental feeding of 50 g/day/cow toxin binder was effective in reducing AFM1 excretion in milk and health risk to consumers.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec13\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eMetabolic Health\\u003c/h2\\u003e \\u003cp\\u003eThe metabolites related to energy-protein nutrition, liver and kidney functions were within normal range and without significant effect of toxin binder, indicating safety of feeding and protecting effect on vital parameters physiological function on body. Similarly, non-significant (P\\u0026thinsp;\\u0026gt;\\u0026thinsp;0.05) changes in serum glucose, total protein, albumin, globulin, SGOT, SGPT, creatinine, BUN concentration reported on toxin binder supplementation (Vieira et al., \\u003cspan citationid=\\\"CR35\\\" class=\\\"CitationRef\\\"\\u003e2024\\u003c/span\\u003e; Gallo et al., \\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e; Guo et al., \\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e) in dairy animals.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec14\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eTreatment Cost\\u003c/h2\\u003e \\u003cp\\u003eThe reduced treatment cost per unit of milk in T2 group, highlight that adding 50 g/day toxin binder was most significant to accrue health benefit, and lowering treatment costs (97.24%).\\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"CONCLUSION\",\"content\":\"\\u003cp\\u003eCows fed total mixed ration and 50 g/day toxin binder (T2) showed positive body weight gain, higher yield of milk, fat-corrected milk, energy-corrected milk, and milk components with maintained milk components level compared to control. Despite greater aflatoxin B1 intake from higher feed consumption, supplementation reduced concentration, daily excretion, bioconcentration, and carry-over of aflatoxin M1in milk by 76.84, 73.18, 75.00, and 76.14%, respectively over control. The toxin binder also not influenced liver, kidney function and energy-protein nutrition. The treatment cost per unit milk production was also reduced by 97.24%, indicating health benefits. The toxin binder at higher dose rate (100 g/day) was without further benefits on measured parameters. Thus, feeding of 50 g/day toxin binder proved effective in enhancing performance and welfare of cow while minimizing aflatoxin health risk to consumers.\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eAcknowledgment\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eWe gratefully acknowledge College of Veterinary Science and Animal Husbandry, Kamdhenu University, Kamdhenu University, Anand, Gujarat, India for providing exceptional support and research facilities that were instrumental in enabling the successful completion of this work.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAuthor contributions\\u003c/strong\\u003e:\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003ePML, BRD, JHP: formulated research; supervise research, HA: executed research, laboratory analysis and data analysis, AMS, HA: Helped in laboratory analysis, wrote manuscript, JHP, DGV: wrote \\u0026amp; revised manuscript. All authors approved the manuscript.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eEthical approval:\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eAn experiment protocol was approved by\\u0026nbsp;Institutional Animal Ethics Committee (IAEC) of College of Veterinary Science \\u0026amp; Animal Husbandry, Kamdhenu University, Anand (Gujarat, India) and cows were cared accordingly. The experiment was conducted at Livestock Research Station,\\u0026nbsp;College of Veterinary Science \\u0026amp; Animal Husbandry,\\u0026nbsp;Kamdhenu University, Anand, Gujarat, India.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConflict of Interest:\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThis was contractual research and funded by JP Veterinary, Mehsana, Gujarat, India.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eData availability\\u003c/strong\\u003e:\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eThe datasets of the current study will available from the corresponding author on reasonable request.\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\n\\u003cli\\u003eAiello, S. E., and Moses, M.A. (2016). 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A., Thurmann, J. P., and Staufenbiel, R. (2020). 24-h variations of blood serum metabolites in high-yielding dairy cows and calves. \\u003cem\\u003eBMC Veterinary Research\\u003c/em\\u003e, 16: 327, 1-11. https://doi.org/10.1186/s12917-020-02551-9\\u003c/li\\u003e\\n\\u003cli\\u003eIARC (2002) International Agency for Research on Cancer; Some traditional herbal medicines, some mycotoxins, naphthalene, and styrene. Summary of data reported and evaluation. \\u003cem\\u003eMonographs on the Evaluation of the Carcinogenic Risk to Humans. \\u003c/em\\u003eInternational Agency for Research on Cancer, Lyon, France, 82\\u003cem\\u003e,\\u003c/em\\u003e 41\\u0026ndash;168. PMID:12687954 \\u003c/li\\u003e\\n\\u003cli\\u003eKutz RE, Sampson JD, Pompeu LB, Ledoux DR, Spain JN, V\\u0026aacute;zquez-A\\u0026ntilde;\\u0026oacute;n M, Rottinghaus GE (2009) Efficacy of Solis, NovasilPlus, and MTB-100 to reduce aflatoxin M1 levels in milk of early to mid-lactation dairy cows fed aflatoxin B1. \\u003cem\\u003eJournal of Dairy Science\\u003c/em\\u003e 92(8): 3959\\u0026ndash;3963. https://doi.org/10.3168/jds.2009-2031 \\u003c/li\\u003e\\n\\u003cli\\u003eLoi M, Fanelli F, Zucca P, Liuzzi V C, Quintieri L, Cimmarusti MT, Monaci L, Haidukowski M, Logrieco AF, Sanjust E, Mul\\u0026egrave; G (2016) Aflatoxin B1 and M1 degradation by Lac2 from \\u003cem\\u003ePleurotus pulmonarius\\u003c/em\\u003e and redox mediators. \\u003cem\\u003eToxins\\u003c/em\\u003e 8(9): 245. https://doi.org/10.3390/toxins8090245 \\u003c/li\\u003e\\n\\u003cli\\u003eLunagariya PM, Parmar CP, Trivedi PG, Desai DM, Agravat PH, Sorathiya KK, Wadhwani KN (2024) Ameliorating effects of Mycotoxcease (Mycotoxin binder) on aflatoxin M1 excretion in dairy cows. \\u003cem\\u003eIndian Journal of Veterinary Science and Biotechnology\\u003c/em\\u003e 20(6): 114\\u0026ndash;119. https://doi.org/10.48165/ijvsbt.20.6.22 \\u003c/li\\u003e\\n\\u003cli\\u003eMaki CR, Thomas AD, Elmore SE, Romoser AA, Harvey RB, Ramirez-Ramirez HA, Phillips TD (2016) Effects of calcium montmorillonite clay and aflatoxin exposure on dry matter intake, milk production, and milk composition. \\u003cem\\u003eJournal of Dairy Science\\u003c/em\\u003e 99(2): 1039\\u0026ndash;1046. https://doi.org/10.3168/jds.2015-9988 \\u003c/li\\u003e\\n\\u003cli\\u003eMehany A, Shams A (2019) Effect of toxin binder on productive performance of lactating Friesian cows. \\u003cem\\u003eJournal of Animal and Poultry Production\\u003c/em\\u003e 10(12): 405\\u0026ndash;413. 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Oxford and IBH Publishing Co., Calcutta, India. DOI: https://doi.org/10.1017/S0021859600074104 \\u003c/li\\u003e\\n\\u003cli\\u003eTolosa J, Rodr\\u0026iacute;guez-Carrasco Y, Ruiz MJ, Vila-Donat P (2021) Multi-mycotoxin occurrence in feed, metabolism and carry over to animal-derived food products: A review. \\u003cem\\u003eFood and Chemical Toxicology\\u003c/em\\u003e 158: 112661. https://doi.org/10.1016/j.fct.2021.112661 \\u003c/li\\u003e\\n\\u003cli\\u003eTyrrell HF, Reid JT (1965) Prediction of the energy value of cow\\u0026rsquo;s milk. \\u003cem\\u003eJournal of Dairy Science\\u003c/em\\u003e 48: 1215\\u0026ndash;1223. https://doi.org/10.3168/jds.S0022-0302(65)88384-9 \\u003c/li\\u003e\\n\\u003cli\\u003eUSFDA (2018) United States Food and Drug Administration, United State of America. Retrieved from https://www.fda.gov/AnimalVeterinary/Products/ucm050223.\\u003cbr\\u003e htm.\\u003c/li\\u003e\\n\\u003cli\\u003eVeldman A, Meijs JAC, Borggreve GJ, Tol JJHD (1992) Carry-over of aflatoxin from cows\\u0026rsquo; food to milk. \\u003cem\\u003eAnimal Science\\u003c/em\\u003e 55(2): 163\\u0026ndash;168. https://doi.org/10.1017/S0003356100037417 \\u003c/li\\u003e\\n\\u003cli\\u003eVieira DJC, Fonseca LM, Poletti G, Martins NP, Grigoletto NTS, Chesini RG, Tonin FG, Cortinhas CS, Acedo TS, Artavia I, Faas J, Takiya CS, Corassin CH, Renn\\u0026oacute; FP (2024) Anti-mycotoxin feed additives: Effects on metabolism, mycotoxin excretion, performance, and total-tract digestibility of dairy cows fed artificially multi-mycotoxin-contaminated diets. \\u003cem\\u003eJournal of Dairy Science\\u003c/em\\u003e 107(10): 7891\\u0026ndash;7903. https://doi.org/10.3168/jds.2023-24539 \\u003c/li\\u003e\\n\\u003cli\\u003eXiong JL, Wang YM, Nennich TD, Li Y, Liu JX (2015) Transfer of dietary aflatoxin B1 to milk aflatoxin M1 and effect of inclusion of adsorbent in the diet of dairy cows. \\u003cem\\u003eJournal of Dairy Science\\u003c/em\\u003e 98(4): 2545\\u0026ndash;2554. https://doi.org/10.3168/jds.2013-7842\\u003c/li\\u003e\\n\\u003c/ol\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":false,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":false,\"isAuthorSuppliedPdf\":false,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":false,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"mycotoxin-research\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"myre\",\"sideBox\":\"Learn more about [Mycotoxin Research](http://link.springer.com/journal/12549)\",\"snPcode\":\"12550\",\"submissionUrl\":\"https://submission.nature.com/new-submission/12550/3\",\"title\":\"Mycotoxin Research\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"stoa\",\"reportingPortfolio\":\"Springer Hybrid\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false},\"keywords\":\"Aflatoxin M1, Body weight, Crossbred cows, Feed safety, Milk yield, Toxin binder\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-9039431/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-9039431/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003eMycotoxin contamination of livestock feed poses a significant threat to animal health, productivity, and food safety. To overcome this, study was conducted to evaluate the effect of toxin binder supplementation on body weight, body condition score, milk yield and composition, and aflatoxin M1 (AFM1) excretion in milk of crossbred lactating cows. Eighteen multiparous cows (75% Holstein Friesian \\u0026times; 25% Kankrej- Indian Zebu), similar in milk yield, days in milk, and lactation numbers were randomly divided into three groups of six each and fed for 70 days. The control group (T1) fed basal total mixed ration (TMR) without toxin binder, and two supplemented groups receiving toxin binder at 50 g/day (T2) and 100 g/day (T3). The toxin binder comprised hydrated sodium calcium aluminosilicate (bentonite), organic acids, mannan oligosaccharides, \\u003cem\\u003eBacillus\\u003c/em\\u003e (direct-fed microbial), oxine copper, and herbal ingredients. Toxin binder supplementation improved (p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.001) milk yield, 4% fat-corrected milk, and energy-corrected milk yields, with T2 showing the highest increase. Milk composition was maintained in T2 but declined at higher dosage (T3). Despite greater aflatoxin B1 intake with higher feed consumption, toxin binder markedly reduced AFM1 concentration and excretion in milk by 76.8\\u0026ndash;78.7% and 73.2\\u0026ndash;75.2%, respectively, compared with control, and lowered bio-concentration and carry-over percentages by about 75\\u0026ndash;78%. The toxin binder also not influenced liver, kidney function and energy-protein nutrition. The treatment cost per kilogram of milk also decreased by 97.2% in T2 group. It was concluded that supplementation of toxin binder at 50 g/day was most effective in improving milk and components yield, and welfare of cow while minimizing aflatoxin health risk to consumers.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Effect of toxin binder feeding on body weight, milk and components yield, and aflatoxin M1 excretion in crossbred lactating cows\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2026-04-16 15:30:02\",\"doi\":\"10.21203/rs.3.rs-9039431/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2026-04-24T21:26:11+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"37163176694408172868672836425830707414\",\"date\":\"2026-04-13T23:19:55+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewersInvited\",\"content\":\"\",\"date\":\"2026-04-08T15:33:51+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorAssigned\",\"content\":\"\",\"date\":\"2026-03-17T02:00:43+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"checksComplete\",\"content\":\"\",\"date\":\"2026-03-17T02:00:42+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"submitted\",\"content\":\"Mycotoxin Research\",\"date\":\"2026-03-05T11:10:18+00:00\",\"index\":\"\",\"fulltext\":\"\"}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"mycotoxin-research\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"myre\",\"sideBox\":\"Learn more about [Mycotoxin Research](http://link.springer.com/journal/12549)\",\"snPcode\":\"12550\",\"submissionUrl\":\"https://submission.nature.com/new-submission/12550/3\",\"title\":\"Mycotoxin Research\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"stoa\",\"reportingPortfolio\":\"Springer Hybrid\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false}}],\"origin\":\"\",\"ownerIdentity\":\"3ea91153-c83c-4aea-83d6-6f67f244d531\",\"owner\":[],\"postedDate\":\"April 16th, 2026\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"under-review\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2026-04-16T15:30:02+00:00\",\"versionOfRecord\":[],\"versionCreatedAt\":\"2026-04-16 15:30:02\",\"video\":\"\",\"vorDoi\":\"\",\"vorDoiUrl\":\"\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-9039431\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-9039431\",\"identity\":\"rs-9039431\",\"version\":[\"v1\"]},\"buildId\":\"XKTyCvWXoU3ODBz1xrDgd\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}