This thesis describes a series of four experiments designed to evaluate the role of the supply of protein in livestock grazing high quality pasture during mating and during pregnancy. The first two studies investigated the effects of high crude protein content of spring or autumn re-growth pasture on the reproductive performance of dairy cows and of ewes at mating. The last two studies investigated how the dietary supply of protein, body condition and their interactions contribute to the breakdown of immunity during the peri-parturient period in ewes and investigated underlying endocrine mechanisms. In the first study (Chapter 3) cows were blood sampled via the tail vein during the breeding period in spring. Plasma was then analysed for urea concentration. Cows with high plasma urea (HPU) or low plasma urea (LPU) were defined as those with plasma urea concentrations of ≥ or < 44.9 mg/dl respectively. Lactating cows (n = 200) were also categorized into high milk producers (HMP) or low milk producers (LPM) relative to an average daily yield of 26.6 l/d. Pasture clipping showed an average pasture CP (crude protein) content of 223 g/kg DM. Concentrations of plasma urea ranged from 26.6 to 64.4 mg/dl. No correlation was observed between plasma urea concentration and either reproductive indicators or milk parameters. Mean blood urea concentration of HPU cows was 50.8 compared to 38.5 mg/dl in LPU cows. There was a trend for more animals (P = 0.09) in the HPU group than in the LPU group not to return to oestrus. Cumulative pregnancy rate in HPU and LPU was similar except at week 6 after the start of mating when more (P < 0.01) HPU than LPU cows were pregnant. Calving to conception interval, calving interval and interval between conception and first service were similar (P > 0.05) between HPU and LPU cows. Gestation length, calving rate, milk yield and milk components were also similar (P > 0.05) between LPU and HPU cows. There was no difference (P > 0.05) in plasma urea concentrations between HMP and LMP milk producers. However, calving to conception interval, interval between calving and first service and calving interval were longer (P < 0.001), submission rate higher (P < 0.001) and NRR (Non-return rate) higher (P < 0.05) in LMP than HMP. The number of services, the interval between first and second service, gestation length and CR (calving rate) were similar (P > 0.05) between HMP and LMP cows. HMP had lower (P < 0.001) milk protein and fat concentrations than LMP cows. This information indicates that, despite the fact that plasma urea was consistently higher than levels in the literature which have been associated with reduced fertility in dairy cows; no impairment of reproductive performance was observed. In the second experiment (Chapter 4) mature and dry Coopworth ewes were blocked by weight, body condition and previous prolificacy (high, HP vs low twinning frequency, LP) into two groups and thereafter randomly allocated to diet which were designed to provided either 1) high protein (163 g/kg DM, ryegrass/red clover pasture, HPP) or low protein (119 g/kg DM, hay and barley grain, HB) supply at joining. These were designed to provide high and low plasma urea concentration. Over a period of 17 days, ewes recorded as mated were examined by laparoscopy, at which time there was no difference in blood urea concentration (58.6 vs 56.1 mg/dl) between HPP and HB groups. Fifty days after the start of joining the number of foetuses present was counted using ultrasonography. As a consequence of lack of difference in the plasma urea concentration, irrespective of treatment group, individual animals were categorized into high (HU) and low plasma urea (LU) status based on whether plasma urea concentration was higher or lower than the sample mean of 51.5 mg urea/dl. Lambs which weighed greater than the mean plus one standard deviation for their litter size were classified as oversize. Ovulation rate and conception rate were similar (P > 0.05) between HPP and BH and between HU and LU ewes. Ewes with previous high reproductive performance (HP) as would be expected had higher ovulation rate (P < 0.001) and conception rate (P < 0.01) than LP ewes. Embryo losses was not (P = 0.06) different between HB and HPP ewes. Urea category (HU vs LU) did not (P > 0.05) influence embryo mortality. Foetal loss, neonatal loss, total reproductive loss and mean lamb birth weight was were not affected by diet, nor urea category (P > 0.05). Single ovulations had tended (P = 0.08) to contribute to higher embryo loss compared to multiple ovulations, and, single foetuses suffered higher (P<0.001) losses compared to multiples. While the study did not achieve large differences in plasma urea concentrations between diets, the levels of plasma urea operating were high yet reproductive wastage rates were similar to those recorded in the literature. Together with similar apparent lack of effect on a high plasma urea environment, the data suggest either that previous findings from controlled studies have a more complex aetiology or that pastoral animals can adapt to high tissue ammonia/urea status. The third trial (Chapter 5) was designed to provide information on the supply of amino acids to the abomasum from protein supplementation which have previously been found to overcome dietary scarcity associated with limitation of peri-parturient increase in FEC. Twin-suckling ewes were fitted with rumen and abomasal cannulae and grazed a ryegrass/clover sward (C) or the same sward but with a 500 g/h/d protein supplement (S). The trial was designed as a cross-over with two 14 day adaptation periods followed by two five-day digesta-sampling periods. All ewes were treated with anthelmintic 14 days after lambing. Weekly analysis of blood glucose was carried on whole blood and analysis of amino acids in plasma. The flows of amino acids (AA) and dry matter (DM) at the abomasum were measured during both sampling periods using intra-ruminally infused markers. Live weight and faecal egg count (FEC) were recorded weekly. Diurnal variation in AA flow at the abomasum peaked between 12:00 and 15:00 h and was greatest in S ewes. Flows of AA, including DAPA, were increased by supplementation by 16%, while sulphur amino acids (SAA) were the most enhanced (by 21%) and flows of leucine, lysine, glutamine and aspartate were increased by about 20%. There were significant time effects in rumen and abomasal pH (P < 0.01; in both cases in both periods) reflecting increase in pH after 09.00 h. During Period II, rumen pH in digesta of C ewes was significantly higher (P < 0.001) than that of S ewes (6.7 ± 0.05 vs 6.4 ± 0.05 for C and S ewes, respectively). Plasma AA concentrations (P < 0.01) were lower in S ewes 21 days after parturition, but similar (P > 0.05) to those of C ewes at other times. Forty-three days after lambing (after cross over), the order was reversed as plasma methionine and cysteine concentrations of C ewes became low (P < 0.05). These changes in plasma AA were accompanied by changes in body condition score between day 23 and 70 post-partum whereby C ewes lost more body condition than S ewes. There was evidence for a lower FEC in S ewes, being 46 vs. 670 epg, respectively for S and C groups (P = 0.08) 21 days after anthelmintic treatment. There were higher (P < 0.05) blood glucose levels in C compared to S ewes at day +35 relative to lambing which was reversed and significantly higher (P < 0.01) for S ewes by day +56 from lambing (after treatments were reversed). There was no significant effect of treatment on live weight and lamb performance. There are limited data in amino acid supply on lactating ewes on pasture and the present study contributes additional information on the supply of amino acids at the abomasum. The prediction that flow rates that sulphur amino acids may have been enhanced to the greatest degree could be significant since sulphur amino acids are needed for the synthesis of glutathione for immune response. It can be calculated that supplementation to supply the quantities of S-amino acid at pasture would be needed, since it would not be possible for sheep to increase pasture intake to achieve similar S-amino acid flow. Increase in bypass amino acids in S ewes at certain times in the day probably suggests influence by protein supplementation at certain times of the grazing cycle. Reduced plasma free amino acids at day +21 relative to lambing, may indicate sparing of body protein breakdown by protein supplementation. However, the difference in blood glucose on day 35 and day 56 may indicate re-adjustment of hormonal settings, responsible for nutrient partitioning. The last study (Chapter 6) used ewes during the peri-parturient period on pasture. Eighty pregnant ewes were allocated into four groups balanced for anticipated number of lambs. Group 1 had a high body condition score (BCS) of 4.0 which was maintained throughout pregnancy by pasture allowance (HM; n = 20). Group 2 (n= 40) had medium body condition (BCS 3.0) and were split into two subgroups; one was offered pasture to allow gain of condition (MH; n = 20) and the second allowed to lose condition by offering a low grazing allowance (ML; n = 20). Group 3 were thin ewes (BCS 2.4) and pasture allowance was designed to maintain this condition (LM; n = 20). These feeding regimes were maintained for 3 weeks from week -8 of pregnancy. During week -5 to -4 all ewes were acclimatized to a protein supplement (60 g/d). A glucose tolerance test (GTT) was conducted during week -4 after which half of the ewes in each group were offered a protein supplement at the rate of 500 g/d, creating –S and –NS groups. During wk -2, a second GTT was carried out. Animals were treated with an anthelmintic 3 wks before lambing, and were then challenged with a dose of 10 000 Teladorsagia circumcincta larvae on weeks -2 and -1 relative to lambing. Weekly recording of FEC, live weight and body condition was carried out. Lambs were weighed within 24 h of birth and again at 44 and 65 d of age. Computed tomography body scanning was carried out on ewes at weeks -8, -3 and +8 relative to lambing. There were no differences (P > 0.05) in lamb performance due to body condition or protein supplementation. FEC of all groups was low (≈ 9 peg) and there was no (P > 0.05) significant difference between ewes of different body condition or due to effects of protein supplementation. Ewes bearing/bearing multiple lambs had the highest FEC at day -32 and +12 relative to lambing, which was significant (P < 0.05) on the latter date. There were no significant effects of supplementation on parasite status. There were differences in basal plasma glucose concentration between groups (P < 0.001), being highest in HM/S and least in ML/NS ewes and was generally higher (P <0.001) during GTT 2 than GTT 1. Ewes carrying a single foetus had higher (P <0.001) basal glucose than those carrying multiple lambs (2.2 vs. 1.7 mmol/L, respectively). Other plasma glucose response indexes were similar (P <0.05) between groups. There were differences in insulin responsiveness between groups (P < 0.001), being highest in MH/S and least in ML/S ewes. Insulin responsiveness tended (P = 0.06) to be lower during GTT 1 than GTT 2, but was higher (P < 0.01) in ewes carrying singles than multiples. There was tendency for higher though non-significant, basal insulin concentrations in HM ewes. Insulin trends over time after glucose infusion suggest greater insulin response at GTT 1. Basal insulin was not correlated with CT muscle weight. Despite differences in body muscle mass at the start of the trial and differences induced by nutrition during late pregnancy, positive gains in muscle mass occurred during early pregnancy and muscle mass was similar in all groups by day 56 of lactation. Animals with greatest fat content at parturition (HM) mobilised the greatest amount and those with least fat (LM) deposited fat during lactation. Further experimentation may consider the use of the hyperinsulinemic-euglycemic clamp approach to more precisely estimate whether hormonal re-setting through insulin resistance may be involved in relaxation of immunity during the peri-parturient period.

This thesis was conducted to evaluate important epidemiological features of Gastro-Intestinal Nematode (GIN) infections in Ontario sheep flocks; namely, the PeriParturient Egg Rise (PPER), overwintering of GIN free-living stages on pasture, and Anthelmintic Resistance (AR). Three main studies were carried out: a longitudinal study was conducted on six sheep farms to evaluate the PPER in ewes lambing in different seasons and to determine whether total plasma protein (TPP) levels and packed cell volume (PCV) were associated with increased fecal GIN-egg shedding. Secondly, a pilot-study was conducted on three farms to describe pasture-level environmental conditions and over-wintering survival and infectivity of free-living GIN larvae, especially Haemonchus contortus. Lastly, a cross-sectional study was conducted on 47 sheep farms in Ontario, to evaluate the frequency of AR, compare different diagnostic tests for AR, and evaluate management practices associated with AR. In the longitudinal study, the PPER was observed in winter, spring and autumn lambing ewes, though the magnitude and distribution of the PPER varied with season. Lower TPP and PCV values were associated with increased fecal GIN-egg counts. The pilot-study suggested that H. contortus larvae did not overwinter successfully on pasture, while other GINs, such as Teladorsagia sp., Trichostrongylus spp. and Nematodirus spp., were able to overwinter on pasture, and were infective the following spring. Resistance to ivermectin, fenbendazole and levamisole was demonstrated on 97% (28/29), 95% (19/20) and 6% (1/17) respectively of the farms tested; most of the resistance observed was found in Haemonchus sp. The Fecal Egg Count Reduction percentage following treatment was influenced by which mean (i.e. arithmetic vs. geometric) was used in the formula; use of pre-treatment in addition to post-treatment faecal egg counts was not influential. Both the fecal egg count reduction test and the larval development assay diagnosed resistance, but there was poor agreement between the two tests, as indicated by the Kappa test. The prior use of benzimidazoles on farms was associated with higher levels of fenbendazole resistance. The information generated in this thesis will be used to develop a parasite control program for sheep flocks in Ontario and to guide future research on GIN parasitism. / New Directions - Ontario Ministry of Agriculture, Food and Rural Affairs; Ontario Sheep Marketing Agency; Ontario Agriculture Centre Canada; Merial; Animal Health Strategic Initiative - University of Guelph Partnership; Undergraduate Research Assistantship; Gartshore Memorial Sheep Scholarship;

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal

Betriebe in Mittelgebirgslagen mit einem hohen Anteil Grassilagefütterung, der damit verbundenen größeren K-Aufnahme sowie höheren DCAD (Dietary-Cation-Anion-Difference) sollten für Gebärparese stärker prädisponiert sein. Ziel dieser Arbeit war es deshalb, Ursachen des Festliegens im Erzgebirge unter Berücksichtigung der DCAD zu ermitteln. Dazu wurden sowohl Fütterungsparameter und die Körperkondition analysiert, als auch Blut- und Harnuntersuchungen ante (a.p.) und post partum (p.p.) durchgeführt. In 21 Betrieben wurden insgesamt 249 Kühe im Zeitraum von 2 Wochen ante partum (a.p.) bis eine Woche p.p. untersucht. Bei 206 gesunden Kühen erfolgte 2 Wochen a.p. eine jeweils einmalige Harn- und Blutentnahmen. Ferner wurden bei 43 peripartal festliegenden Kühen nur Blut- und Harnproben direkt vor der Behandlung analysiert. Die sonographische Ermittlung der Rückenfettdicke (RFD) erfolgte bei je zehn klinisch gesunden Kühen pro Betrieb je 2 Wochen a.p. Die Silageproben (Grundfutteranalyse) wurden hinsichtlich ihrer Inhaltsstoffe untersucht und die DCAD berechnet. Laktationsnummern, Milchleistungen und Krankheitsinzidenzen der Kühe wurden ermittelt. Im Blutserum wurden ß-Hydroxy-Butyrat (BHB), Cholesterol, Bilirubin, Aspartat-Aminotransferase (ASATS), Creatinkinase (CK), Alkalische Phosphatase (AP), Protein (TP), Albumin, Harnstoff, Creatinin, Calcium (Ca), anorganisches Phosphat (Pi), Magnesium (Mg), Natrium (Na), Kalium (K) und im Vollblut die Leukozytenanzahl bestimmt. Im Harn erfolgte die Analyse derselben Mineralstoffe wie im Serum sowie weiterhin des pH-Wertes, der NSBA (Netto-Säure-Basen-Ausscheidung) und des Creatinins. Errechnet wurden die fraktionierten Eliminationen der einzelnen Mineralstoffe, der BSQ (Basen-Säure-QuotienTS) aus den Mengen der Basen, Säuren und Ammoniumionen im Harn sowie der Creatininquotient aus Harn- und Serumcreatinin. Befunde der Klinisch gesunden Kühe zwei Wochen a.p.: Die durchschnittliche RFD der Kühe lag in den Betrieben im Mittel zwischen 19 bis 26 mm. Muskel- und Knochenstoffwechsel charakterisierende Parameter waren physiologisch. Die Mediane der AP-Aktivität im Serum lagen in 15 der 21 Betriebe unter 100 U/l. Energie- und Eiweißstoffwechsel, Leukozytenanzahlen sowie Mineralstoffe in Serum und Harn ergaben keine Hinweise für präpartale Stoffwechselstörungen. Die Mediane von pH-Wert, NSBA sowie K- und Creatininkonzentrationen im Harn überschritten in mindestens fünf Betrieben die Grenzwerte, da die K-Konzentrationen in den Silagen bis fünffach über dem empfohlenen Bereich lagen. Die DCAD lag im Mittel aller Betriebe mit 458 meq/kg T weit über –100 bis +150 meq/kg T. Eine gesicherte Korrelation zur Anzahl der Festlieger bestand nicht. Befunde der Festlieger p.p.: Die Festlieger hatten zu je 60% reduzierte Ca- sowie Pi- und zu 16% verminderte Mg-Konzentrationen. Die Verteilung auf die Gebärparesetypen war wie folgt: Typ I = 42% Typ II= 19%, Typ III = 5%, Typ IV = 12% und Typ V = 21%. Dies entspricht mehrheitlich den Literaturangaben. Die Ca-, Pi- und Mg-Konzentrationen im Harn lagen bei den Festliegern in den physiologischen Bereichen. Die Mehrzahl der Festlieger hatte gleichzeitig erhöhte CK- (74%) und AST-Aktivitäten (53%). In 38% der Fälle war die AP-Aktivität vermindert. Die K- und NSBA-Konzentrationen im Harn der Festlieger zeigten als Folge der reduzierten Futteraufnahme gegenüber den Befunden a.p. eine Abnahme bis in den unteren physiologischen Bereich und z.T. darunter. Störungen des Energiestoffwechsels in Form erhöhter Ketonkörper- (58%) und Bilirubin- (40%) sowie reduzierter Cholesterol- (58%) Konzentrationen hatte die Mehrheit der festliegenden Kühe. Dabei war die kombinierte Ca-Pi-BHB-Stoffwechselstörung die Regel, traf aber nicht in jedem Fall zu. Die moderat veränderten Parameter des Energiestoffwechsels sind offensichtlich Folge der reduzierten Futteraufnahme. Eine Steigerung der Leukozytenzahl als Hinweis auf eine akut-entzündliche Reaktion bestand bei 44% der Festlieger, - die Leukopenie war die Ausnahme. Mit der Leukozytose waren in 36,8% respektive 42,1% der Fälle eine Hypokalz- resp. Hypophosphatämie gekoppelt. Bei dem größeren Teil (47,4%) der Leukozytosen waren die Ca- und Pi-Konzentrationen jedoch physiologisch. Das zeigt in Übereinstimmung mit den klinischen Befunden, dass entzündliche Prozesse und Verletzungen als weitere Ursache des Festliegens angesehen werden mussten. Fazit: Die erhöhte DCAD war in den untersuchten Mittelgebirgsbetrieben kein prädisponierender Faktor für die Gebärparese. Neben den bekannten Ca-Pi-Störungen spielten entzündliche Prozesse dafür eine wichtige Rolle.

Betriebe in Mittelgebirgslagen mit einem hohen Anteil Grassilagefütterung, der damit verbundenen größeren K-Aufnahme sowie höheren DCAD (Dietary-Cation-Anion-Difference) sollten für Gebärparese stärker prädisponiert sein. Ziel dieser Arbeit war es deshalb, Ursachen des Festliegens im Erzgebirge unter Berücksichtigung der DCAD zu ermitteln. Dazu wurden sowohl Fütterungsparameter und die Körperkondition analysiert, als auch Blut- und Harnuntersuchungen ante (a.p.) und post partum (p.p.) durchgeführt. In 21 Betrieben wurden insgesamt 249 Kühe im Zeitraum von 2 Wochen ante partum (a.p.) bis eine Woche p.p. untersucht. Bei 206 gesunden Kühen erfolgte 2 Wochen a.p. eine jeweils einmalige Harn- und Blutentnahmen. Ferner wurden bei 43 peripartal festliegenden Kühen nur Blut- und Harnproben direkt vor der Behandlung analysiert. Die sonographische Ermittlung der Rückenfettdicke (RFD) erfolgte bei je zehn klinisch gesunden Kühen pro Betrieb je 2 Wochen a.p. Die Silageproben (Grundfutteranalyse) wurden hinsichtlich ihrer Inhaltsstoffe untersucht und die DCAD berechnet. Laktationsnummern, Milchleistungen und Krankheitsinzidenzen der Kühe wurden ermittelt. Im Blutserum wurden ß-Hydroxy-Butyrat (BHB), Cholesterol, Bilirubin, Aspartat-Aminotransferase (ASATS), Creatinkinase (CK), Alkalische Phosphatase (AP), Protein (TP), Albumin, Harnstoff, Creatinin, Calcium (Ca), anorganisches Phosphat (Pi), Magnesium (Mg), Natrium (Na), Kalium (K) und im Vollblut die Leukozytenanzahl bestimmt. Im Harn erfolgte die Analyse derselben Mineralstoffe wie im Serum sowie weiterhin des pH-Wertes, der NSBA (Netto-Säure-Basen-Ausscheidung) und des Creatinins. Errechnet wurden die fraktionierten Eliminationen der einzelnen Mineralstoffe, der BSQ (Basen-Säure-QuotienTS) aus den Mengen der Basen, Säuren und Ammoniumionen im Harn sowie der Creatininquotient aus Harn- und Serumcreatinin. Befunde der Klinisch gesunden Kühe zwei Wochen a.p.: Die durchschnittliche RFD der Kühe lag in den Betrieben im Mittel zwischen 19 bis 26 mm. Muskel- und Knochenstoffwechsel charakterisierende Parameter waren physiologisch. Die Mediane der AP-Aktivität im Serum lagen in 15 der 21 Betriebe unter 100 U/l. Energie- und Eiweißstoffwechsel, Leukozytenanzahlen sowie Mineralstoffe in Serum und Harn ergaben keine Hinweise für präpartale Stoffwechselstörungen. Die Mediane von pH-Wert, NSBA sowie K- und Creatininkonzentrationen im Harn überschritten in mindestens fünf Betrieben die Grenzwerte, da die K-Konzentrationen in den Silagen bis fünffach über dem empfohlenen Bereich lagen. Die DCAD lag im Mittel aller Betriebe mit 458 meq/kg T weit über –100 bis +150 meq/kg T. Eine gesicherte Korrelation zur Anzahl der Festlieger bestand nicht. Befunde der Festlieger p.p.: Die Festlieger hatten zu je 60% reduzierte Ca- sowie Pi- und zu 16% verminderte Mg-Konzentrationen. Die Verteilung auf die Gebärparesetypen war wie folgt: Typ I = 42% Typ II= 19%, Typ III = 5%, Typ IV = 12% und Typ V = 21%. Dies entspricht mehrheitlich den Literaturangaben. Die Ca-, Pi- und Mg-Konzentrationen im Harn lagen bei den Festliegern in den physiologischen Bereichen. Die Mehrzahl der Festlieger hatte gleichzeitig erhöhte CK- (74%) und AST-Aktivitäten (53%). In 38% der Fälle war die AP-Aktivität vermindert. Die K- und NSBA-Konzentrationen im Harn der Festlieger zeigten als Folge der reduzierten Futteraufnahme gegenüber den Befunden a.p. eine Abnahme bis in den unteren physiologischen Bereich und z.T. darunter. Störungen des Energiestoffwechsels in Form erhöhter Ketonkörper- (58%) und Bilirubin- (40%) sowie reduzierter Cholesterol- (58%) Konzentrationen hatte die Mehrheit der festliegenden Kühe. Dabei war die kombinierte Ca-Pi-BHB-Stoffwechselstörung die Regel, traf aber nicht in jedem Fall zu. Die moderat veränderten Parameter des Energiestoffwechsels sind offensichtlich Folge der reduzierten Futteraufnahme. Eine Steigerung der Leukozytenzahl als Hinweis auf eine akut-entzündliche Reaktion bestand bei 44% der Festlieger, - die Leukopenie war die Ausnahme. Mit der Leukozytose waren in 36,8% respektive 42,1% der Fälle eine Hypokalz- resp. Hypophosphatämie gekoppelt. Bei dem größeren Teil (47,4%) der Leukozytosen waren die Ca- und Pi-Konzentrationen jedoch physiologisch. Das zeigt in Übereinstimmung mit den klinischen Befunden, dass entzündliche Prozesse und Verletzungen als weitere Ursache des Festliegens angesehen werden mussten. Fazit: Die erhöhte DCAD war in den untersuchten Mittelgebirgsbetrieben kein prädisponierender Faktor für die Gebärparese. Neben den bekannten Ca-Pi-Störungen spielten entzündliche Prozesse dafür eine wichtige Rolle.