Oral supplementation of clay to dairy cattle has been reported to reduce toxicity of aflatoxin (AF) in contaminated feed. The objective of this study was to determine the effects of 3 concentrations of dietary clay supplementation in response to an AF challenge. Ten multiparous rumen-cannulated Holstein cows [body weight (mean ± SD) = 669 ± 20 kg and 146 ± 69 d in milk], were assigned to 1 of 5 treatments in a randomized replicated 5 × 5 Latin square design balanced to measure carryover effects. Periods (21 d) were divided in an adaptation phase (d 1 to 14) and a measurement phase (d 15 to 21). From d 15 to 17, cows received an AF challenge. The challenge consisted of 100 μg of aflatoxin B (AFB)/kg of dietary dry matter intake (DMI). The material was fitted into 10-mL gelatin capsules and administered into the rumen through a rumen-cannula based on the average DMI obtained on d 12 to 14. Treatments were no clay plus an AF challenge (POS); 3 different concentrations of clay (0.5, 1, or 2% of dietary DMI) plus an AF challenge; and a control consisting of no clay and no AF challenge (C). Statistical analysis was performed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC). Two contrasts, CONT1 (POS vs. C) and CONT2 (POS vs. the average of 0.5, 1, and 2% clay), were compared along with the linear and quadratic treatment effects (POS, 0.5%, 1%, 2%). Cows supplemented with clay had lower AF excretion in milk as aflatoxin M (AFM; 0.5% = 20.83 μg/d, 1% = 22.82 μg/d, and 2% = 16.51 μg/d) and AF transfer from rumen fluid to milk (AFM 0.5% = 1.01%, 1% = 0.98%, and 2% = 0.74%) compared with cows in POS (AFM = 27.81 μg/d and AF transfer = 1.37%, CONT2). Similarly, concentrations of AFM in milk (0.5% = 0.35 μg/kg, 1% = 0.30 μg/kg, 2% = 0.25 μg/kg), AFB in feces (0.5% = 1.79 μg/g, 1% = 1.52 μg/kg, 2% = 1.48 μg/kg), and AFB in rumen fluid (0.5% = 0.05 μg/kg, 1% = 0.02 μg/kg, 2% = 0.02 μg/kg) were reduced in cows fed clay compared with POS (0.43 μg/kg, 2.78 μg/kg, and 0.10 μg/kg, respectively, CONT2). Cows supplemented with clay tended to have lower 3.5% fat-corrected milk [0.5% = 38.2 kg, 1% = 39.3 kg, 2% = 38.4 kg, standard error of the mean (SEM) = 1.8] than cows in POS (41.3 kg; SEM = 1.8; CONT2). Plasma superoxide dismutase (SOD) concentration tended to be lower for cows fed clay in the diet (0.5% = 2.16 U/mL, 1% = 1.90 U/mL, 2% = 2.3 U/mL; SEM = 0.3) than for cows in POS (2.72 U/mL; CONT2). Additionally, when cows were exposed to AF without clay in the diet, plasma concentrations of aspartate aminotransferase (AST) decreased from 84.23 (C) to 79.17 (POS) and glutamate dehydrogenase (GLDH) decreased from 91.02 (C) to 75.81 (POS). In conclusion, oral supplementation of clay reduced the transfer of AF from the rumen to milk and feces.

Oral supplementation of clay has been reported to function as buffer in dairy cows. However, its effects on rumen, blood, and fecal pH have varied among studies. Our objective was to determine the effects of 3 concentrations of dietary clay supplementation after a grain challenge. Ten multiparous rumen-cannulated Holstein cows [body weight (mean ± standard deviation)=648±12kg] with 142±130 (60 to 502) days in milk were assigned to 1 of 5 treatments in a replicated 5×5 Latin square design balanced to measure carryover effects. Periods (21d) were divided into an adaptation phase (d 1 to 18, with regular total mixed ration fed ad libitum) and a measurement phase (d 19 to 21). Feed was restricted on d 18 to 75% of the average of the total mixed ration fed from d 15 to 17 (dry matter basis), and on d 19 cows received a grain challenge. The challenge consisted of 20% finely ground wheat administered into the rumen via a rumen cannula, based on the average dry matter intake obtained on d 15 to 17. Treatments were POS (no clay plus a grain challenge), 3different concentrations of clay (0.5, 1, or 2% of dietary dry matter intake), and control (C; no clay and no grain challenge). Statistical analysis was performed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC). Contrasts 1 (POS vs. C) and 2 (POS vs. the average of 0.5, 1, or 2%) were compared, along with linear and quadratic treatment effects. Rumen, fecal, and blood pH, along with blood metabolites, were measured at 0, 4, 8, 12, 16, 20, 24, 36, and 48h relative to the grain challenge. Cows fed POS had lower rumen pH [(mean ± standard error) 6.03±0.06] than cows fed C (6.20±0.06). Cow fed POS had lower fecal pH (6.14±0.04) than cows fed C (6.38±0.04). We observed a linear treatment effect for rumen pH and fecal pH. Fecal pH (6.22±0.04) was higher for cows fed clay (contrast 2) then for cows fed POS (6.14±0.04). We also observed a treatment difference (contrast 2) for negative incremental area under the curve, pH below 5.6 × h/d, (0.5% clay=7.93±0.83, 1% clay=8.56±0.83, and 2% clay=7.79±0.83) compared with POS (11.0±0.83). Cows fed clay tended to have higher milk yield (0.5% clay=28.8±3.4kg, 1% clay=30.2±3.4kg, and 2% clay=29.1±3.4kg, contrast 2), and had higher 3.5% fat-corrected milk (0.5% clay=29.9±3.5kg, 1% clay=34.1±3.5kg, and 2% clay=33.1±3.4kg), and higher energy-corrected milk (0.5% clay=29.1±3.3kg, 1% clay=32.8±3.4kg, and 2% clay=31.6±3.3kg) than cows fed POS (27.7±3.4kg, 28.0±3.4kg, 27.7±3.3kg, respectively). In conclusion, cows fed clay had higher rumen pH, energy-corrected milk, fat-corrected milk, and a trend for milk yield than cows fed POS.

The etiology of cystic ovarian follicles (COF) remains a conundrum with steroidogenic, immunological, and metabolic dysfunctions linked to its development. Studies suggest that COF development may occur as a result of disruption of the insulin signaling pathway and the severity of a negative energy balance in dairy cows, but mid to late lactation cows diagnosed with COF are unlikely to have issues with energy metabolism. Herein, we characterized the mRNA expression of steroidogenic (LHCGR, StAR, CYP11A1, 3β-HSD, CYP19A), immunological (IL-1β, IL-6, IL-8, TLR-4, TNF), and metabolic markers (IGF-1, IRS1) in follicular fluid; and plasma and follicular fluid levels of E2, IL-1β, glucose, and NEFA in early and mid-late lactation COF cows. Lactating dairy cows were diagnosed as having COF (n = 11, follicle >20 mm persistent for 7 days, absence of corpus luteum, and flaccid uterus) while 11 herdmates cycling with a dominant follicle were classified as the control cows. Cows diagnosed with COF were classified as early lactation (COF-E, n = 5) cows, <35 days in milk (DIM); or mid-late lactation (COF-M/L, n = 6), ≥118 DIM cows. Results revealed that mRNA expression StAR was greater (P < 0.01) in COF-E cows than COF-M/L cows and the control cows. The mRNA expression CYP19A1 was lower (P < 0.01) in COF-E cows and COF-M/L cows than in the control cows. The mRNA expression IL-6 and IRS-1 tended to be greater and lower, respectively, in COF-M/L cows compared to the control cows. The mRNA expression IGF-1 was greater (P < 0.01) in COF-E and COF-M/L cows than in the control cows. The plasma and follicular fluid concentration of NEFA was greater (P < 0.05) in COF-E cows than in COF-M/L and the control cows. Cows with COF-E had disturbances in steroidogenic and metabolic markers, while cows with COF-M/L had steroidogenic, immunological, and metabolic dysregulations, suggesting that COF pathogenesis may vary between early and mid-late lactation dairy cows.