View Full Table | Close Full ViewTable 1.

Ergovaline and lolitrem B concentrations in ryegrass hay1 and effect on BW and milk production in ewes. Values are expressed as mean ± SD (n = 8)

 
Parameter SE–
SE+
Day 1 Day 8 Day 22 Day 1 Day 8 Day 22
Ergovaline,2 μg/kg <5 <5 <5 1,006 ± 397 674 ± 33 872 ± 351
Lolitrem B,2 μg/kg <10 <10 <10 1,027 ± 139 824 ± 28 800 ± 301
Feed intake,3 kg/ewe∙d 1.35 1.23 1.52 1.53 1.12 1.46
BW, kg 81 ± 10 79 ± 10 80 ± 10 80 ± 9 77 ± 8 79 ± 9
Milk production, L/d 2.6 ± 0.6 2.3 ± 0.6 1.8 ± 0.6† 2.6 ± 0.5 2.2 ± 0.4 1.8 ± 0.6†
Fat content, g/L 49 ± 10 63 ± 6† 61 ± 10† 50 ± 7 66 ± 5† 63 ± 6†
Protein content, g/L 51 ± 4 47 ± 5 51 ± 4 52 ± 4 49 ± 6 51 ± 6
1SE– = endophyte-free ryegrass; SE+ = endophyte-infected ryegrass.
2Mean ± SD concentration measured at the pen level for 1 kg of hay taken from 5 different locations of the bales.
3Mean consumption calculated by dividing the consumption measured at the pen level by the number of ewes (n = 8).
Significant difference from Day 1 (Wilcoxon, P < 0.05).



View Full Table | Close Full ViewTable 2.

Biochemical parameters in the plasma of ewes fed endophyte-free ryegrass (SE–) or endophyte-infected ryegrass (SE+). Values are expressed as mean ± SD (n = 8)

 
Parameter1 SE–
SE +
Day 0 Day 28 Day 0 Day 28
Hematocrit, % 33.5 ± 3.2 33.5 ± 4.4 34.5 ± 1.5 34.8 ± 1.8
Sodium, mEq/L 145 ± 3 146 ± 7 144 ± 3 146 ± 9
Potassium, mEq/L 4.61 ± 0.47 4.56 ± 0.62 4.70 ± 0.45 4.43 ± 0.63
Protein, g/L 74.3 ± 3.4 75.0 ± 7.2 73.7 ± 7.7 77.7 ± 4.5
Urea, mmol/L 7.23 ± 0.79 11.3 ± 1.5† 7.21 ± 1.42 9.75 ± 1.61†
Creatinine, μmol/L 68.9 ± 3.8 71.3 ± 10.4 64.2 ± 7.62 68.7 ± 8.8
Cholesterol, mmol/L 2.10 ± 0.37 1.78 ± 0.54† 2.20 ± 0.34 1.93 ± 0.32†
Triglycerides, mmol/L 0.25 ± 0.05 0.15 ± 0.02† 0.23 ± 0.03 0.18 ± 0.05†
Total bilirubin, μmol/L 2.00 ± 0.00 2.00 ± 0.00 2.00 ± 0.00 2.00 ± 0.00
AST, UI/L 155 ± 48 134 ± 23 162 ± 27 174 ± 50
ALT, UI/L 20.2 ± 8.3 21.2 ± 6.3 22.7 ± 3.6 25.6 ± 7.7
CPK, UI/L 150 ± 20 135 ± 39 177 ± 49 170 ± 43
LDH, UI/L 1,093 ± 284 1,006 ± 144 1,154 ± 211 1,140 ± 102
1AST = aspartate aminotransferase; ALT = alanine aminotransferase; CPK = creatine phosphokinase; LDH = lactate dehydrogenase.
Significant difference from Day 0 (Wilcoxon, P < 0.05).



View Full Table | Close Full ViewTable 3.

Effects of 28 d of feeding ewes endophyte-free ryegrass (SE–) or endophyte-infected ryegrass (SE+) on oxidative stress parameters. Values are expressed as mean ± SD (n = 8)

 
Parameter1 Day 28
SE– SE+
Plasma
    MDA, nmol/(min ∙ mg of plasma proteins) 7.92 ± 1.03 8.81 ± 1.21
    SOD, milliunits/(min ∙ mg of plasma proteins) 6.00 ± 0.74 7.37 ± 0.97*
    CAT, ng CH2O/(min ∙ mg of plasma proteins) 152 ± 96 130 ± 18
    GR, μmol NADPH/(min ∙ mg of plasma proteins) 0.45 ± 0.10 0.46 ± 0.09
    GPx, μmol NADPH/(min ∙ mg of plasma proteins) 0.79 ± 0.22 0.76 ± 0.27
    TGSH, pmol/mg of plasma proteins 3.44 ± 0.88 3.42 ± 0.86
Liver
    MDA, nmol/mg of cytosolic proteins 62.6 ± 23.6 52.4 ± 7.6
    MDA, mmol/mg of microsomal proteins 6.34 ± 1.42 6.80 ± 0.42
    SOD, units/(min ∙ mg of cytosolic proteins) 14.4 ± 3.4 12.3 ± 2.2
    CAT, μg CH2O/(min ∙ mg of cytosolic proteins) 506 ± 30 472 ± 32
    GR, μmol NADPH/min/(min ∙ mg of cytosolic proteins) 19.5 ± 4.1 21.4 ± 2.6
    GPx, μmol NADPH/min/(min ∙ mg of cytosolic proteins) 101 ± 8 108 ± 9
    TGSH, μmol/mg of cytosolic proteins 20.2 ± 11.2 17.5 ± 6.7
    GSSG, μmol/mg of cytosolic proteins 1.29 ± 0.71 1.40 ± 1.14
Kidneys
    MDA, nmol/mg of cytosolic proteins 76.8 ± 32.4 77.4 ± 29.8
    MDA, mmol/mg of microsomal proteins 13.4 ± 4.22 8.52 ± 1.78
    SOD, units/(min ∙ mg of cytosolic proteins) 11.74 ± 2.56 14.36 ± 2.30*
    CAT, μg CH2O/(min ∙ mg of cytosolic proteins) 67.3 ± 7.4 85.3 ± 15.0*
    GR, μmol NADPH/min/mg of cytosolic proteins 75.5 ± 13.1 84.6 ± 22.9
    GPx, μmol NADPH/(min ∙ mg of cytosolic proteins) 250 ± 36 270 ± 42
    TGSH, μmol/mg of cytosolic proteins 0.66 ± 0.14 0.78 ± 0.12
    GSSG, μmol/mg of cytosolic proteins 0.03 ± 0.02 0.04 ± 0.03
1MDA = malondialdehyde; SOD = superoxide dismutase; CAT = catalase; GR = glutathione reductase; GPx = glutathione peroxidase; TGSH = total glutathione; GSSG = oxidized glutathione; CH2O = formaldehyde; NADPH = nicotinamide adenine dinucleotide phosphate.
*Significant difference between groups (Student, P < 0.05).



View Full Table | Close Full ViewTable 4.

Effects of 28 d of feeding ewes endophyte-free ryegrass (SE–) or endophyte-infected ryegrass (SE+) on hepatic and renal drug metabolizing enzymes. Values are expressed as mean ± SD (n = 8)

 
Parameter Day 28
SE– SE+
Liver, g 1,246 ± 349 1,015 ± 388
    O-dealkylation1
        Ethoxyresorufin 4.28 ± 1.92 4.24 ± 1.54
        Methoxyresorufin 1.82 ± 0.28 2.50 ± 0.44*
        Pentoxyresorufin 1.54 ± 0.42 1.52 ± 0.18
    N-demethylation2
        Aminopyrine 112 ± 27 97 ± 25
        Benzphetamine 101 ± 28 111 ± 18
        Dimethylnitrosamine 25.2 ± 4.1 23.9 ± 5.7
        Erythromycin 21.4 ± 8.2 18.3 ± 6.7
        Ethylmorphine 54.9 ± 11.0 48.4 ± 7.6
    Glutathione S-transferase
        CDNB3 1.02 ± 0.22 0.86 ± 0.14
        DCNB4 1.30 ± 0.28 1.16 ± 0.26
    UDPGT5
        PNP6 1.22 ± 0.20 1.16 ± 0.20
Kidneys, g 199 ± 41 204 ± 25
    O-dealkylation1
        Ethoxyresorufin 1.70 ± 0.24 1.34 ± 0.46
        Methoxyresorufin 2.90 ± 0.86 1.74 ± 0.30*
        Pentoxyresorufin 0.40 ± 0.28 0.50 ± 0.20
    Glutathione S-transferase
        CDNB3 0.24 ± 0.02 0.26 ± 0.04
        DCNB4 0.30 ± 0.12 0.12 ± 0.04*
    UDPGT5
        PNP6 0.20 ± 0.08 0.41 ± 0.06*
1Expressed in picomoles resorufin formed/(minute∙milligram of microsomal proteins).
2Expressed in nanograms formaldehyde formed/(minute∙milligram of microsomal proteins.
3CDNB = 1-chloro-2,4-dinitrobenzene; expressed in millimoles of 1-chloro-2,4-dinitrobenzene conjugated/(minute∙milligram of cytosolic proteins).
4DCNB = 1,2-dichloro-4-nitrobenzene; expressed in micromoles of 1,2-dichloro-4-nitrobenzene conjugated/(minute∙milligram of cytosolic proteins).
5UDPGT = uridine diphosphate glucuronosyltransferase.
6PNP = para-nitrophenol; expressed in micromoles of paranitrophenol conjugated/(minute∙milligram of microsomal proteins).
*Significant difference between groups (Student, P < 0.05).



View Full Table | Close Full ViewTable 5.

Ergovaline and lolitrem B and concentrations in tissues and milk in ewes fed with endophyte-infected ryegrass after 28 d of exposure

 
Matrix Ergovaline
Lolitrem B
n < LD1 LD < n < LQ >LQ2 Mean ± SD3 Maximum3 n < LD1 LD < n < LQ >LQ4 Mean ± SD3 Maximum3
Milk 4 4 0 6 2 0
Liver 0 3 5 0.68 ± 0.15 0.86 4 2 2 0.71 ± 0.15 0.81
Kidneys 0 6 2 0.53 ± 0.04 0.56 7 0 1 0.6
Muscle 8 0 0 0 4 4 0.77 ± 0.17 0.95
Fat 6 2 0 0 0 8 2.39 ± 1.67 4.78
Brain 8 0 0 8 0 0
1LD = limit of detection: 0.15 μg/L or 0.15 μg/kg for milk and tissues, respectively.
2LQ = limit of quantitation: 0.7 μg/L for milk, 0.8 μg/kg for fat and brain, and 0.5 μg/kg for liver, kidneys, and muscle.
3In micrograms/liter or micrograms/kilogram for milk and tissues, respectively.
4LQ = 0.5 μg/L or 0.5 μg/kg for milk and tissues, respectively.