In der vorliegenden Arbeit wurden die Ergebnisse von 263 Einzeltierverfolgungen in 12 Betrieben in verschiedenen Regionen in Deutschland, Belgien, Frankreich und der Tschechischen Republik zur Stallkokzidiose der Kälber zusammengefasst ausgewertet. Während diesen Untersuchungen wurden 5840 Kotproben beurteilt. Dabei wurden das Auftreten und die Ausprägung der Eimeriose der Kälber analysiert und potentielle Einflussfaktoren untersucht und ein verlässlicher Schwellenwert für die Bewertung der Oozystenausscheidung ermittelt. Weiterhin wurden in 16 Betrieben bei 23 gemeinsam aufgestallten Kälbergruppen Spezies-Prävalenzuntersuchungen über einen Zeitraum von fünf Wochen durchgeführt. Hierfür wurden 5133 Proben ausgewertet, davon 3519 mit Teil- und 1614 mit vollständiger Differenzierung. In allen Betrieben traten die Kotkzidiosen als eine Mischinfektion verschiedener Eimeria-Arten auf. Dabei herrschte in der Regel in jedem Betrieb eine der beiden pathogensten Spezies vor. Diese Prädominanz blieb auch über mehrere Jahre gleich in den Betrieben. Die Herkunft der Kälber hatte einen Einfluss auf den Infektionsverlauf. In Betrieben mit ausschließlich eigener Nachzucht verläuft die Kokzidiose als eingipflige Infektion, bei Zukaufbetrieben ist der Infektionsverlauf zweigipflig. Ein signifikant erhöhtes Risiko des Auftretens einer klinisch ausgeprägten Kokzidiose bei Aufstallung auf Stroheinstreu im Vergleich zur Haltung auf Spaltenboden konnte bewiesen werden (p = 0,005). In der Verfolgungsuntersuchung korrelierten die Kokonsistenzwerte mit den nachgewiesenen OpG in den Kotproben positiv signifikant (0,135 Korrelationskoeffizient; p = 0,000). Für das Auftreten von Durchfall konnte eine statistisch bewiesene lineare Korrelation mit der Oozystenausscheidung für diverse Schwellenwerte festgestellt werden (p = 0,000). Bei einem Grenzwert von 500 OpG lag der Korrelationskoeffizient bei 0,149. Die Korrelationswerte erhöhten sich nicht wesentlich bei Festlegung eines höheren Grenzwertes für die Oozystenausscheidung oder sanken sogar (0,153 bis 0,121). Bei der Verrechnung von Durchfallvorkommen mit gleichzeitiger, potentiell relevanter Oozystenausscheidung mit den verschiedenen Schwellenwerten der Oozystenausscheidung konnte der höchste Zusammenhang zwischen dem Durchfallgeschehen und dem Schwellenwert von 500 OpG bewiesen werden (0,633 Korrelationskoeffizient; p = 0,000). Daher kann ein Wert von 500 OpG pathogener Eimeria spp. als geeigneter Schwellenwert für die Feststellung einer relevanten Oozystenausscheidung angesehen werden. Bei gleichzeitigem Auftreten von Durchfall ist von einer maßgeblichen Beteiligung der Eimerien auszugehen. Ein gehäuftes Auftreten von mit Oozystenausscheidung assoziierten Durchfällen trat bei Tieren ohne oder ohne potentiell relevante Oozystenausscheidung (‚rK -’) signifikant seltener auf (p = 0,000) als bei Tieren mit mindestens 500 aufsummierten OpG während des gesamten Beobachtungszeitraumes (‚rK +’). Kälber der Kategorie ‚Kok-Kat 1’ hatten signifikant niedrigere Kotkonsistenzwerte und weniger Durchfälle als Tiere der Subpopulation ‚rK -’ (p = 0,000). Dagegen hatten die Tiere der Auswertungsgruppe ‚Kok-Kat 2’ statistisch bewiesen in allen Durchfall-Kategorien höhere Werte bzw. ein häufigeres Durchfallvorkommen als beide anderen Subpopulationen (p = 0,000 für alle Vergleiche). Wässrige Durchfälle mit Beimengungen traten, mit Ausnahme einer Einzelbeobachtung in Gruppe ‚rK -’, ausschließlich bei Kälbern der Auswertungsgruppe ‚Kok-Kat 2’ auf. Sowohl Kälber der Gruppe ‚rK +’ als auch ‚Kok-Kat 2’-Tiere (jeweils p = 0,000) und Kälber mit potentiell relevanter Oozystenausscheidung, aber ohne gleichzeitiges Durchfallgeschehen, (‚Kok-Kat 1’) (p = 0,005) hatten signifikant geringere relative Gewichtszunahmen als Tiere ohne bzw. ohne potentiell relevante Oozystenausscheidung (‚rK -’). Eine lineare Korrelation der Ausscheidung der pathogenen Spezies E. bovis und E. zuernii mit den absoluten (-0,098 Korrelationskoeffizient; p = 0,005) und relativen Gewichtszunahmen (-0,170 Korrelationskoeffizient; p = 0,000) konnte statistisch bewiesen werden. Bei Haltung auf Stroheinstreu zeigten Tiere ohne bzw. mit weniger als 500 ausgeschiedenen OpG im Untersuchungszeitraum (‚rK -’) signifikant höhere relative Zunahmen im Vergleich zu Tieren mit potentiell relevanter Oozystenausscheidung (p = 0,000). Dabei war es ohne Bedeutung, ob diese Kälber ein gleichzeitiges Durchfallgeschehen zeigten oder nicht. Besonders zum Tragen kommen diese Unterschiede in der Gewichtsentwicklung in den Wochen mit den höchsten Oozystenausscheidungen bei zweigipfligem Infektionsverlauf. Unter den Milchviehkälbern nahmen Tiere ohne relevante Oozystenausscheidung signifikant mehr relatives Gewicht zu als Kälber mit kumulativ mindestens 500 OpG im Untersuchungszeitraum (p = 0,004). Dies galt sowohl für Tiere mit gleichzeitigem Durchfall ‚Kok-Kat 2’ (p = 0,002) als auch tendenziell für Kälber der Gruppe ‚Kok-Kat 1’ (p = 0,059). Mastviehkälber der Gruppe ‚rK -’ zeigten signifikant höhere relative Zunahmen im Vergleich zu Tieren mit potentiell relevanter Oozystenausscheidung ‚rK +’ (p = 0,039). Dies galt auch in Relation zu den Kälbern der Auswertungsgruppe ‚Kok-Kat 1’ (p = 0,029). Während der Prävalenzuntersuchungen wurden insgesamt neun verschiedene Eimeria-Arten nachgewiesen. In Einzelkotproben wurden zwischen einer und neun verschiedene Spezies beobachtet. Unter den in Europa als heimisch bekannten Arten wurden während dieser Untersuchung lediglich E. wyomingensis, E. brasiliensis und E. bukidnonensis nicht gefunden. E. ellipsoidalis hatte sowohl die höchste Inzidenz (20,99 %) als auch die größte Intensität (arithmetischer Mittelwert von 765963,37 OpG), gefolgt von E. bovis und E. zuernii. Zudem wurde in Aufzuchtbetrieben E. ellipsoidalis in der Regel als erste Eimeria-Spezies, gefolgt von E. auburnensis und den pathogenen Arten E. zuernii und E. bovis, nachgewiesen. Die seltensten Spezies waren E. canadensis und E. pellita. Eimeria pellita wurde als letzte Art erst ab der fünften Woche nach Aufstallung beobachtet. E. cylindrica trat vermehrt in den Betrieben in Belgien und Frankreich auf. Diese Art sowie E. canadensis wurden nur in Betrieben in Bayern, Belgien und Frankreich festgestellt. E. pellita hatte, neben E. canadensis, die geringsten Prävalenzen, Nachweise wurden vor allem für zentral gelegene Betriebe sowie im Süden des Untersuchungsgebietes dokumentiert. Die Übereinstimmung der gefundenen Varianzen der Speziesprävalenzen der vorliegenden Untersuchung mit den Daten epidemiologischer Studien in den verschiedenen Regionen aus der Literatur bestätigt einen repräsentativen Charakter der Untersuchungsbetriebe. Dies belegt zusätzlich die Allgemeingültigkeit der festgestellten Einflussfaktoren auf das Auftreten und die Auswirkungen der Stallkokzidiose der Kälber. / A total number of 263 calves housed on 12 different farms in several regions in Germany, Belgium, France and the Czech Republic were followed individually in tracking studies and data was compiled and analysed to investigate factors influencing occurrence and severity of bovine eimeriosis of housed calves. The same data was used for development and verification of a suitable threshold indicating relevant oocyst excretion. Within the tracking studies a total number of 5840 faecal samples were examined for faecal consistency, oocyst counts of pathogenic E. bovis and E. zuernii excretion carried out and individual body weight development was documented regularly. Additionally 23 groups of animals on 16 farms were observed for a period of five weeks and 5133 faecal samples examined for oocyst excretion and Eimeria species differentiated. All study sites were positive for mixed coccidia species infections. Nevertheless all farms except one showed a predominance of one pathogenic Eimeria species, which remained unchanged in different groups of animals and even in different years of investigations. Animal origin, i.e. groups of animals representing own breeding or originating of only one source in contrast to groups of calves coming from several origins, influences the course of the coccidiosis infection. Farms with only one single and permanent animal origin or raising exclusively the own breeding show coccidiosis with a single peak of oocyst excretion. On farms housing groups of animals of various origins the course of infection and oocyst shedding has typically two peaks with an interval of two to three weeks. The risk for development of clinical coccidiosis rises significantly if animals were housed on straw bedding compared to slatted-floor (p = 0.005). Faecal scores correlated significantly (p = 0.000) with the intensity of oocyst excretion with a positive correlation coefficient of 0.135. For occurrence of diarrhoea a positive linear correlation with the oocyst excretion was statistically proved (p = 0.000) for various thresholds. At a threshold of 500 opg of E. bovis and E. zuernii the correlation coefficient rised to 0.149 and correlation coefficients did not rise distinctly or even got down if higher thresholds were used (values between 0.153 and 0.121). Focusing only on potentially coccidiosis related diarrhoea the threshold of 500 opg of E. bovis and E. zuernii proved to result in the highest correlation (0.633; p = 0.000) of all tested threshold levels. Therefore the threshold of 500 opg of E. bovis and E. zuernii can be accounted modest and reliable to detect a relevant oocyst excretion in individual faecal samples as well as in compiled samples. In cases of coincidental diarrhoea coccidiosis can be considered as a major factor. Increased numbers of days with diarrhoea in coincidence with an oocyst excretion (‘Kokass-DF’) within the observation period were seen significantly more often (p = 0.000) in animals with a potentially relevant oocyst excretion (‘rK +’) of at least 500 summed up opg of E. bovis and E. zuernii in comparison to calves without such an oocyst excretion (‘rK -‘). Significantly lower faecal scores and fewer days with diarrhoea were documented for calves of the group ‘Kok-Kat 1’ in contrast to animals of group ‘rK -‘ (p = 0.000). Nevertheless significantly higher faecal scores and more days with diarrhoea than both other groups were calculated for those calves meeting the inclusion criteria for group ‘Kok-Kat 2’ (p = 0.000 for all comparisons). Additionally liquid faeces or faeces with constituencies were seen only in this group, except for one single sample of a calf of group ‘rK -‘. Calves of evaluation group ‘rK +’ as well as both subpopulations representing group ‘Kok-Kat 2’ and calves with potentially relevant oocyst excretion but without diarrhoea associated to an oocyst excretion (‘Kok-Kat 1’) showed significant lower values for relative body weight increases in comparison to animals without relevant oocyst excretion throughout the complete study period of five weeks (‘rK -‘) (p = 0.000 vs. ‘rK +’ and vs. ‘Kok-Kat 2‘; p = 0.005 compared with ‘Kok-Kat 1’). A negative linear correlation between oocyst excretion of pathogenic Eimeria spp. and absolute (-0.098 correlation coefficient; p = 0.005) as well as relative body weight gain (-0.170 correlation coefficient) was verified statistically (p = 0.000). Animals housed on straw bedding and belonging to the group ‘rK -‘ gained relatively more body weight in comparison to calves housed in the same stables and meeting the inclusion criterium of group ‘rK +’, i.e. excreting at least 500 summed up opg of pathogenic E. spp., (p = 0.000) within the total study period. The presence of coincidental diarrhoea had no impact on impaired body weight development of animals with a potentially relevant coccidia excretion. Differences in body weight development were most distinct within the weeks of highest intensities in oocyst excretion according to a course of infection with two peaks. Within the subpopulation of dairy calves those animals belonging to evaluation group ‘rK -‘ developed significantly higher relative body weight gains compared to group ‘rK +’ (p = 0.004). Similar results were found for animals of group ‘Kok-Kat 2’ (p = 0.002) and a statistical tendency was calculated for group ‘Kok-Kat 1’ (p = 0.059) in comparison to group ‘rK -‘. Analogous to the differences in dairy calves animals on fattening farms without relevant oocyst excretion (‘rK -’) had significantly higher relative body weight gains compared to calves of evaluation group ‘rK +’ (p = 0.039) and animals of group ‘Kok-Kat 1’ (p = 0.029) of the same breeds and farms. Nine different Eimeria spp. were detected during the prevalence studies. In single individual samples a minimum of one and up to nine different species were found. Twelve Eimeria spp. are known to be endemic in Europe of which only E. brasiliensis, E. bukidnonensis and E. wyomingensis were not present in any faecal sample in this study. E. bovis and E. zuernii were only second to E. ellipsoidalis which had the highest prevalence (20.99 %) as well as the highest intensity (765963.37 mean opg) in the faecal samples examined. In breeding farms E. ellipsoidalis was the first species to be found in faecal examination in most cases, followed by E. auburnensis and the pathogenic species E. zuernii and E. bovis. E. canadensis and E. pellita were detected only in a low number of samples. E. pellita was observed for the first time at the faecal samplings five weeks after stabling and mainly in farms situated in the central and southern region of the prevalence study. E. canadensis and E. cylindrica were most prominent in farms situated in Belgium and France. Variances in prevalence of the species observed are in conformity with those to be found in recent literature according to the different regions of Europe. This may indicate a representative character of the farms participating in this study and therefore universal validity of the results and impactfactors on coccidiosis in calves described in this manuscript.

Diarrhoe ist eines der häufigsten Probleme beim equinen Neonaten. Nahezu alle Fohlen entwickeln Durchfall innerhalb der ersten Lebenswochen. Unterschiedliche virale, bakterielle und parasitäre Ursachen werden diskutiert. In diesen Zeitraum fällt ebenfalls die erste Rosse der Stute, sodass der Durchfall um den 5. - 15. Lebenstag (LT) bei den Fohlen als „Fohlenrossedurchfall“ bezeichnet wird. Es wird vermutet, dass die Entwicklung der intestinalen Mikroflora und die Reifung der Darmschleimhaut im Wesentlichen für das Durchfallgeschehen verantwortlich sind. Bisher ist jedoch wenig bekannt über die Entwicklung der intestinalen Mikroflora bei Fohlen. Einige Probiotika sind als Darmflorastabilisatoren bei Tieren zugelassen. Studien belegten positive Effekte von Toyocerin® (B. cereus var. toyoi) auf die Darmgesundheit bei anderen Tierarten wie z.B. Kälbern, Ferkeln, Broilern, Puten und Mastkaninchen. Die vorliegende Arbeit sollte klären, ob die Supplementierung von B. cereus var. toyoi zu einer Stabilisierung der sich entwickelnden intestinalen Mikroflora und damit zu einer Verringerung der Durchfälle bei Fohlen führt. Die Untersuchung wurde an 25 Mutterstuten eines Vollblutgestüts und ihren Fohlen durchgeführt. Alle Fohlen wurden von Februar bis Mai 2011 geboren. Von Geburt an wurden die Fohlen randomisiert in drei Behandlungsgruppen eingeteilt: Placebo-Gruppe (10 ml isotone Kochsalzlösung, n=8), 50 mg Toyocerin-Gruppe (5 x 108 KbE B. cereus var. toyoi gelöst in 10 ml isotoner Kochsalzlösung, n=7) und 200 mg Toyocerin-Gruppe (2 x 109 KbE B. cereus var. toyoi gelöst in 10 ml isotoner Kochsalzlösung, n=10). Die Placebo- und Behandlungsgruppen wurden einmal täglich vom 1. – 58. LT supplementiert. Herz- und Atemfrequenz, Körperinnentemperatur und die Körpermasseentwicklung wurden nach einem standardisierten Protokoll erhoben. Kotproben konnten mit Hilfe von Kotsammelbeuteln oder durch rektale Entnahme innerhalb von 24 Stunden nach der Geburt sowie an LT 9, 16, 23, 30, 44, 58 und am ersten Durchfalltag gewonnen werden. Blutproben wurden aus der Vena jugularis externa am 1., 9., 16., 30., 58. LT sowie am ersten Durchfalltag entnommen. Die bakteriologische Untersuchung erfolgte mit Hilfe des Kulturverfahrens. Die Bestimmung der Gesamt-IgG-Werte wurde mit einem kompetitiven ELISA, die Bestimmung der spezifischen Antikörper IgG-anti-LPS von E. coli J5 und IgG-anti-PLC-von-C. perfringens-1a mit einem indirekten ELISA durchgeführt. 88 % der Fohlen entwickelten Durchfall (Placebo 7/8, 50 mg Toyocerin 5/7, 200 mg Toyocerin 10/10) mit einer hohen Inzidenz zwischen dem 8. und 16. LT. Das Allgemeinbefinden und die Bewegungs- und Sauglust blieben dabei unbeeinflusst. Zum Zeitpunkt des ersten Östrus der Stute zeigten 59 % der Fohlen Diarrhoe. Unter den 41 %, die keinen Durchfall zum Zeitpunkt der Fohlenrosse hatten, waren Fohlen, die nie Durchfall vom 1. – 58. LT zeigten, aber auch welche die Diarrhoe entwickelten, als die Mutterstute sich nicht in Rosse befand. Ein Zusammenhang zwischen der Fohlenrosse der Stute und Durchfall bei ihrem Fohlen konnte nicht hergestellt werden. Es zeigte sich eine Tendenz, dass hohe Spiegel der Gesamt-IgG (>20 mg/ml) und IgG-anti-LPS von E. coli J5 (>120 RE/ml) nach der Kolostrumaufnahme im Zusammenhang mit einer geringeren Anzahl von Durchfalltagen innerhalb der ersten zwei Lebensmonate standen. C. perfringens und Enterobakterien waren gleichermaßen nachweisbar bei Fohlen mit Durchfall als auch bei unauffälligen Fohlen. Aus der Supplementierung von B. cereus var. toyoi ergab sich kein Effekt auf die Kotflora der Fohlen, außer auf die Gesamtkeimzahlen (GKZ) der aeroben Bakterien. Bei den Aerobiern im Fohlenkot konnte ein signifikanter Behandlungseffekt (p=0,012) festgestellt werden. Im ersten Milchkot der Fohlen waren GKZ von 4,5 x 104 KbE/g (200 mg Toyocerin-Gruppe) bis 5,0 x 105 KbE/g (50 mg Toyocerin-Gruppe) bei den aeroben Bakterien und GKZ von 2,4 x 105 KbE/g (200 mg Toyocerin-Gruppe) bis 4,7 x 106 KbE/g (50 mg Toyocerin-Gruppe) median bei den Anaerobiern nachweisbar. Danach stieg der Gehalt der aeroben und anaeroben Bakterien weiter bis zum 3. LT und stagnierte bis zum 16. LT. Während dieser Stagnationsphase trat bei 92 % der Fohlen (23/25) eine Veränderung der Kotkonsistenz bis hin zu Durchfällen auf. Vom 16. bis zum 58. LT sanken die Gehalte moderat bei den Aerobiern median am 58. LT auf 2,7 x 105 KbE/g (Placebo-Gruppe) bis 2,2 x 106 KbE/g (50 mg Toyocerin-Gruppe) und bei den Anaerobiern median am 58. LT auf 3,8 x 105 KbE/g (Placebo-Gruppe) bis 2,9 x 106 KbE/g (200 mg Toyocerin-Gruppe). Bis zum 58. LT näherte sich der Medianwert der aeroben und anaeroben Bakterien im Kot der Placebo-Gruppe dem Wert der Mutterstuten (gemessen am ersten Tag nach der Geburt) an. Innerhalb der ersten Lebenstage war eine hohe aerobe sowie anaerobe Keimzahl im Kot der Fohlen nachzuweisen, die sich oberhalb der Keimzahlen befand, die im Kot der Mutterstuten zum Zeitpunkt der Geburt gemessen wurde. Im Rahmen der Entwicklung und Etablierung der bakteriellen intestinalen Mikroflora wurde das Fohlenrossedurchfallgeschehen bei den Fohlen beobachtet. B. cereus var. toyoi hatte dabei keinen Einfluss auf die Anzahl der Fohlen mit Durchfall und den Gesundheitsstatus der Fohlen. / Diarrhoea is probably one of the most common problems in equine neonates. Almost all foals develop transient diarrhoea within the first weeks of life. Different viral, bacterial and parasitic causes are discussed. Between the 5th and the 15th day of the foal’s life, when their dam’s first post partum (p.p.) oestrus is expected, diarrhoea in foals is observed quite often. That is why it’s called “foal heat diarrhoea”. In literature establishment of intestinal microflora and maturation of the intestinal mucosa is responsible for the occurrence of diarrhoea in this period of life. But little is known about the development of the intestinal microflora in foals. Many probiotics are authorised as gut flora stabilisers in animal nutrition. Some studies proved positive effects of Bacillus (B.) cereus var. toyoi (Toyocerin®) on intestinal health in other species e.g. calves, piglets, broiler chicken, poultry and growing rabbits. The present study deals with the question if a supplementation of B. cereus var. toyoi lead to a stabilisation of the developing intestinal microflora and therefore to a reduction of diarrhoea in foals. A total of 25 mares and foals of a thoroughbred stud were included into the study. Foals were born between February and May 2011. From birth, the foals were randomly assigned to three treatment groups: placebo group (10 ml isotonic saline solution, n=8), 50 mg Toyocerin group (5 x 108 cfu B. cereus var. toyoi solved in 10 ml isotonic saline solution, n=7) and 200 mg Toyocerin group (2 x 109 cfu B. cereus var. toyoi solved in 10 ml isotonic saline solution, n=10). Placebo- and treatment groups were orally supplemented once a day starting on the 1st through to the 58th day of life. Determination of heart and respiratory rate, body temperature, body weight was realised according to a standardised protocol. Within the first day of life, on day 9, 16, 23, 30, 44, 58 and on the first day of diarrhoea faecal samples has been taken from the rectum or by the use of a collection bag. Blood samples were taken via jugular venipuncture on day 1, 9, 16, 30, 58 and on the first day of diarrhoea. Culture-depend methods were used to analyse the bacterial microflora. Serum IgG was analysed by a competitive ELISA, IgG-anti-LPS from E. coli J5 and IgG-anti-PLC-from-C. perfringens-1a by an indirect ELISA. 88 % of the foals developed diarrhoea (placebo 7/8, 50 mg Toyocerin 5/7, 200 mg Toyocerin 10/10) with a high incidence between the 8th and the 16th day of the foal’s life. Meanwhile, foals remained bright and alert and continued to nurse. At the time point of the first p.p. oestrus in the mares, 59 % of their foals showed signs of diarrhoea. Within the remaining 41 % there are foals that had no diarrhoea but there are also foals which had diarrhoea when the mare had not been in heat. Neonatal diarrhoea in foals is not linked to p.p. oestrus in their mares. There was a tendency, that high serum-IgG (> 20 mg/ml) and IgG-anti-LPS from E. coli J5 (> 120 RE/ml) after colostrum uptake were associated with lower diarrhoea severity in the first 58 days of the foal’s life. C. perfringens and enterobacteria can be found equally in foals with diarrhoea and in foals which are not afflicted. B. cereus var. toyoi supplementation had no effect on faecal bacteria in foals, except on aerobic bacteria (p=0,012). In the first milk faeces aerobic bacteria were detected in median from 4,5 x 104 cfu/g (200 mg Toyocerin-group) to 5,0 x 105 cfu/g (50 mg Toyocerin-group) and anaerobic bacteria were detected in median from 2,4 x 105 cfu/g (200 mg Toyocerin-group) to 4,7 x 106 cfu/g (50 mg Toyocerin-group). Afterwards the counts increased towards the 3rd day of life and stayed on a high level till the 16th day of life. During this stagnation in 92 % of the foals a change in faecal consistency and diarrhoea was observed. Afterwards, from the 16th though to the 58th day of life, the bacteria counts in the faeces moderately decreased in median for the aerobic bacteria on the 58th day of life down to 3,8 x 105 cfu/g (placebo-group) till 2,9 x 106 cfu/g (200 mg Toyocerin-group). On the 58th day of life the counts of aerobic and anaerobic bacteria in the faeces of the placebo-group approached the counts in the faeces of the mare (measured at the time point of birth). In the first days of foals’ life detection of aerobic and anaerobic bacteria in the faeces were high, and above the level of the bacteria counts in the faeces of the mare at the time point of birth. Foal heat diarrhoea is observed as a part of the development and establishment of bacterial intestinal microflora. B. cereus var. toyoi had no effect on the percentage of foals with diarrhoea and health status in the foals at that point.

Diarrhoea and related diseases are the most common causes of death in children, especially from developing countries, killing about 1.5 million children under the age of five yearly. In South Africa, diarrhoea is the third leading cause of death. This condition results from food and water sources infected with Campylobacter spp., Escherichia coli, Salmonella spp., Shigella spp., Giardia intestinalis and Cryptospondium parvum amongst others. Diarrhoea spreads through faeces-contaminated water. Hence, infection is more common when there is a short supply of clean drinking and cooking water. Waterborne diseases are common in rural communities of Bizana because the majority of rural dwellers depend largely on water from unprotected sources. Most of the pathogens that cause diarrhoea have developed resistance to several antibiotics. Therefore there is a need for new and safe antidiarrhoeal drugs. Most people in developing countries use traditional medicine to treat all kinds of diseases including diarrhoea and South Africa is no exception. Each cultural group in South Africa has different medical solutions for the prevention and curing of the same disease. The people from Pondoland (AmaMpondo), around Bizana have a strong tradition of using medicinal plants for the cure and prevention of several conditions including diarrhoea. Although several researchers have conducted different types of studies in many parts of South Africa to evaluate the efficacy of traditional medicine used in the treatment of diarrhoea, there is, however, still a lot of undisclosed data that should be collected. The aims of this research were to record and collect medicinal plants that are used for treating diarrhoea in Bizana, Pondoland in the Eastern Cape and evaluate them for their pharmacological properties. An ethnobotanical approach is one of several methods that have been useful in selecting plants for pharmacological research, yielding better results than other plant selection methods. Using questionnaires, this approach was used to record plants that are used for treating diarrhoea in Bizana for testing in pharmacological assays. From the completed questionnaires, nine plants were selected for bioassays based on their higher frequency index, and the fact that the plants have never been evaluated against diarrhoea causing-microorganisms. The study revealed 34 plant species belonging to 21 families as being used in treatment of diarrhoea in the study area. Psidium guajava was the most mentioned plant species. The dried, ground plant materials were each extracted non-sequentially using petroleum ether (PE), dichloromethane (DCM), 70% ethanol (EtOH) and water. Among all the extracts, 70% ethanol yielded the highest quantity of crude extract. The extracts were each evaluated for their antibacterial, anti-inflammatory and genotoxicity properties. For the antibacterial activity, the following diarrhoea causing microorganisms were used: Gram-positive Staphylococcus aureus and Gram-negative Campylobacter jejuni, E. coli and Shigella flexneri. A microdilution assay (for S. aureus, E. coli and S. flexneri) and the disk diffusion technique (for C. jejuni) were used for antibacterial testing. The extracts were also evaluated for their ability to inhibit cyclooxygenase (COX-1 and -2) enzymes. Genotoxicity was evaluated using the Salmonella microsome assay without S9 metabolic activation. Three strains of Salmonella typhimurium TA98, TA1535 and TA1537 were used. The evaluated plant extracts showed a broad spectrum of inhibitory activity with MIC values ranging from 0.098-12.5 mg/ml and mean zone inhibition percentage ranging from 0-73%. The best activity was exhibited by DCM extracts of Rapanea melanophloeos, EtOH extracts of Ficus craterostoma and Maesa lanceolata with MIC values of 0.098 mg/ml and EtOH extracts of Searsia chirindensis with 73% mean zone inhibition percentage. The inhibitory activity against COX-1 enzyme was higher than COX-2, with 19 plant extracts for the former and 7 for the latter. The highest inhibition of COX-1 was shown by EtOH extracts of F. craterostoma and the DCM extract of S. chirindensis at 100%. Highest percentage COX-2 inhibition was shown by water extracts of F. craterostoma and DCM extracts of Tecoma capensis with 99.5% and 99.0% respectively. None of the tested plant extracts were mutagenic, at all concentrations tested against all tester strains of the bacteria. The results of this study demonstrate that people still have a rich and diverse pool of knowledge concerning the uses of plants against diarrhoea. The data also show that plants form part of the cultural heritage of the communities in Pondoland. Therefore it is important to urgently save the people’s cultural heritage by recording the existing knowledge and confirming therapeutic uses of plants through scientific methods. This will prevent the information from vanishing together with the ageing knowledge holders. In light of the fact that the evaluated plants were selected based on their ethnobotanical use for treating diarrhoea, the activities reported here goes a long way in adding value to the plants used as part of traditional medicine. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2012.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2002. / Härtill 6 uppsatser.

To determine the relationship between infant-feeding practices and diarrhoeal infections, a descriptive survey was conducted to infants between six to 12 months of age. A guided interview was conducted to 105 mothers of infants who attended the health facilities of Mbabane, Swaziland. The results show that breast-feeding is routinely practiced by the majority of mothers and exclusive breast-feeding is very low, but supplementary feed in the form of formula or solids are introduced by the majority of respondents within the first three months of life. Infants who were given colostrum and breast milk had fewer diarrhoeal attacks. Other factors, for example education and cultural factors influenced the feeding practices and number of diarrhoeal attacks. It is recommended that breast-feeding should be promoted as an important intervention in the control of diarrhoea / Health Studies / M.A. (Nursing Science)

Made available in DSpace on 2014-06-11T19:27:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-02-26Bitstream added on 2014-06-13T20:35:33Z : No. of bitstreams: 1 alexandrino_b_me_jabo.pdf: 222632 bytes, checksum: 48c6532830e7cdbb18fc1060e2cf6feb (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O presente trabalho teve como objetivo verificar a variação da ocorrência da Rinotraqueíte Infecciosa Bovina (IBR) pela associação com duas doenças virais imunossupressoras: a Diarréia Viral Bovina (BVD) e a Leucose Enzoótica Bovina (LEB), em seis propriedades onde não se adota esquema de vacinação contra essas enfermidades. Amostras de soro sangüíneo foram analisadas no teste de virusneutralização (VN), para constatação de IBR e BVD, e Imunodifusão em Gel de Ágar (IDGA), para a LEB. Foram selecionados cinco rebanhos bovinos, em propriedades localizadas em municípios dos Estados de São Paulo e Minas Gerais, sendo três de exploração leiteira, um de gado de corte e um misto, com animais soropositivos ao BoHV-1, além de um rebanho controle, sem anticorpos contra essa enfermidade. Das 278 amostras analisadas, 54,68% (152/278) foram positivas ao BoHV-1, 69,70% (194/278) ao BVDV-1 e 34,33% (96/278) ao VLEB. Na análise estatística, ao relacionar cada enfermidade com o tipo de exploração do rebanho e a idade dos animais, houve diferença significativa, indicando que estas variáveis são fatores de risco para as enfermidades estudadas. Em relação ao tipo de exploração, os rebanhos leiteiros foram mais suscetíveis ao BoHV-1 e a LEB (81,31% e 49,53% respectivamente, (α = 1 ) enquanto no rebanho de gado de corte o BVDV-1 teve maior ocorrência (94,74%, α = 1). A idade foi fator de risco apenas para o BoHV-1 e a LEB, sendo os animais mais velhos os mais suscetíveis (α = 1). As associações entre o BoHV-1 e o BVDV-1, e o BoHV-1 e a LEB também foram significativas (α = 5 e α = 1 respectivamente), indicando que em rebanhos infectados por BVDV-1 e/ou LEB, a probabilidade de se encontrar o BoHV-1 é maior do que naqueles onde não ocorre essas duas enfermidades. / The present research had as objective to verify the variation of the occurrence of Infectious Bovine Rhinotracheitis (IBR) by association with two viral infections that affect the immune system, Bovine Viral Diarrhoea (BVD) and enzootic bovine leukosis (EBL), in six farms where vaccination against these diseases was not adopted. Serum samples had been analyzed by the virus neutralization (VN) test for IBR and BVD diagnosis, and agar gel immunodiffusion (IDGA) test for EBL diagnosis. Five cattle herds with BoHV-1 seropositive animals had been selected in the states of São Paulo and Minas Gerais, three of them exploiting dairy cattle, one exploiting beef cattle and one exploiting mixed cattle, in addition to a control herd without seropositive animals. From 278 analyzed samples, 54.68% (152/278) reacted to the BoHV-1, 69.70% (194/278) to the BVDV-1, and 34.33% (97/278) to the EBLV. The statistic analysis showed a significant difference (α = 1) in infection occurrence according to the kind of exploitation and the age of the animals. Dairy cattle were more sensitive to the BoHV-1 (81.31%) and to the EBLV (49.53%) and less to BVDV-1 infection (45.79%). Among the beef herds, the major occurrence was BVDV-1 infection (94.74%), followed by BoHV-1(34.19%) and EBLV (3.95%). The age was a risk factor (α = 1) only for BoHV-1 and EBLV. The associations between BoHV-1 and BVDV-1 infections (α = 5) and between BoHV-1 and EBLV infections (α = 1) also indicated that among BVDV-1 and/or EBLV infected herds the probability of finding BoHV-1 is higher than among herds where these two infections does not occur.

Department of Animal Science / MSCAGR (Animal Science)

MSc (Microbiology) / Department of Microbiology / Background: Escherichia coli is a facultative anaerobic bacterium that forms part of the gut microbiota. It is used as an indicator that confirms recent faecal contamination. E. coli have been identified amongst the pathogens that are mostly responsible for moderate to severe diarrheal outbreaks in the low and middle-income countries. With South Africa facing an issue in water scarcity, issues concern poor sanitation and hygiene practices results in serious public health problems and allows E. coli to be transmitted from infected human or animal faeces to a new susceptible host using environmental reservoirs such as soil, water, hands as the transmission pathway. Objective: The primary objective of the study was to characterize E. coli strains from rural communities of Vhembe district, Limpopo, South Africa. Methodology: Households of 7 villages in the Vhembe district were randomly selected. A total of 81 households (HHs) were part of the study. In each household, a structured questionnaire was used to background information on WASH practices. Samples taken from each HH included toilet seat swabs, floor swabs, child and mother handwash samples, stored water samples and running tap water samples. A total of 399 samples were analysed using Colilert® Quanti-trays®/2000 method to detect the presence of Escherichia coli. Positive E. coli samples were further identified using multiplex polymerase chain reaction (m-PCR) to determine the pathogenic strains of E. coli. Transmission pathways were established using identified strains. Results: Data from the structured questionnaires showed common problems of availability of running tap water; lack of provision of sanitation; open practice on defaecation and very little hand hygiene practices. A total of 91 (22.81%) samples tested positive for E. coli with the Colilert® Quanti-trays®/2000 method. The mothers’ handwash samples had the most E. coli prevalence followed by stored water samples. The most prevalent E. coli pathotype was EPEC with the virulence gene eae. Atypical EPEC (60.44%) outnumbered the typical EPEC (5.49%). The pathotype ETEC was detected in 41.76% samples and EHEC in 9.89% samples. Transmission pathway was observed from the different households; with eae gene (aEPEC) being the most detected from samples looking at the LT gene (ETEC). v | P a g e Discussion: All 7 villages are facing common issues such as lacking running water, poor sanitation and improper hand hygiene practices. The mothers were the most contaminated and it was observed that its due to the daily activities that they perform around the house. It is of importance for them to practice proper hand hygiene to prevent transmission of pathogenic E. coli to the children via direct or indirect transmission route. The pathogenic E. coli was detected from all different samples collected from the households including the floor and toilet seat samples. EPEC was detected the most, and studies have shown that this strain is known to cause diarrheal infections in young children from developing countries. Conclusion: The members of the study village households were aware of the WASH services and its importance. However, proper implementation into their day-to-day life is lacking due to the high number of TC and E. coli detected from handwash samples and stored water samples from the households. Recommendation: Appropriate WASH strategies should be established to ensure good health at the village households. Further studies should be done to check possible transmission pathways from more village households. / NRF

Background: Almost all foals develop transient diarrhoea within the first weeks of life. Studies indicated different viral, bacterial, and parasitic causes, such as rotavirus, Clostridium perfringens, Escherichia coli, and Cryptosporidium are discussed. But little is known about the development of intestinal microflora in foals. The present study investigated whether the supplementation with Bacillus cereus var. toyoi would modify the developing intestinal microflora and consequently reduce diarrhoea in foals. From birth, the foals were randomly assigned to three treatment groups: placebo (10 mL isotonic NaCl, n = 8), low dosage (LD; 5 × 108 cfu B. cereus var. toyoi, n = 7) and high dosage (HD; 2 × 109 cfu B. cereus var. toyoi, n = 10). Treatment groups were supplemented orally once a day for 58 days. Faeces scoring and sampling were performed within the first 24 h after birth and on day 9, 16, 23, 30, 44, 58 of the foal’s life and also on the first day of diarrhoea. Culture-plate methods were used to analyse the bacterial microflora. Results: Eighty-eight per cent of the foals developed diarrhoea (placebo 7/8, LD 5/7, HD 10/10) during the first 58 days of life. Bacillus cereus var. toyoi supplementation had no effect on bacterial microflora. Clostridium perfringens and enterobacteria were equally prevalent in foals with diarrhoea and those who were not afflicted. Conclusions: We conclude that the supplementation of B. cereus var. toyoi had no effect on the occurrence of diarrhoea and health status in the foals.

Diarrhoe ist eines der häufigsten Probleme beim equinen Neonaten. Nahezu alle Fohlen entwickeln Durchfall innerhalb der ersten Lebenswochen. Unterschiedliche virale, bakterielle und parasitäre Ursachen werden diskutiert. In diesen Zeitraum fällt ebenfalls die erste Rosse der Stute, sodass der Durchfall um den 5. - 15. Lebenstag (LT) bei den Fohlen als „Fohlenrossedurchfall“ bezeichnet wird. Es wird vermutet, dass die Entwicklung der intestinalen Mikroflora und die Reifung der Darmschleimhaut im Wesentlichen für das Durchfallgeschehen verantwortlich sind. Bisher ist jedoch wenig bekannt über die Entwicklung der intestinalen Mikroflora bei Fohlen. Einige Probiotika sind als Darmflorastabilisatoren bei Tieren zugelassen. Studien belegten positive Effekte von Toyocerin® (B. cereus var. toyoi) auf die Darmgesundheit bei anderen Tierarten wie z.B. Kälbern, Ferkeln, Broilern, Puten und Mastkaninchen. Die vorliegende Arbeit sollte klären, ob die Supplementierung von B. cereus var. toyoi zu einer Stabilisierung der sich entwickelnden intestinalen Mikroflora und damit zu einer Verringerung der Durchfälle bei Fohlen führt. Die Untersuchung wurde an 25 Mutterstuten eines Vollblutgestüts und ihren Fohlen durchgeführt. Alle Fohlen wurden von Februar bis Mai 2011 geboren. Von Geburt an wurden die Fohlen randomisiert in drei Behandlungsgruppen eingeteilt: Placebo-Gruppe (10 ml isotone Kochsalzlösung, n=8), 50 mg Toyocerin-Gruppe (5 x 108 KbE B. cereus var. toyoi gelöst in 10 ml isotoner Kochsalzlösung, n=7) und 200 mg Toyocerin-Gruppe (2 x 109 KbE B. cereus var. toyoi gelöst in 10 ml isotoner Kochsalzlösung, n=10). Die Placebo- und Behandlungsgruppen wurden einmal täglich vom 1. – 58. LT supplementiert. Herz- und Atemfrequenz, Körperinnentemperatur und die Körpermasseentwicklung wurden nach einem standardisierten Protokoll erhoben. Kotproben konnten mit Hilfe von Kotsammelbeuteln oder durch rektale Entnahme innerhalb von 24 Stunden nach der Geburt sowie an LT 9, 16, 23, 30, 44, 58 und am ersten Durchfalltag gewonnen werden. Blutproben wurden aus der Vena jugularis externa am 1., 9., 16., 30., 58. LT sowie am ersten Durchfalltag entnommen. Die bakteriologische Untersuchung erfolgte mit Hilfe des Kulturverfahrens. Die Bestimmung der Gesamt-IgG-Werte wurde mit einem kompetitiven ELISA, die Bestimmung der spezifischen Antikörper IgG-anti-LPS von E. coli J5 und IgG-anti-PLC-von-C. perfringens-1a mit einem indirekten ELISA durchgeführt. 88 % der Fohlen entwickelten Durchfall (Placebo 7/8, 50 mg Toyocerin 5/7, 200 mg Toyocerin 10/10) mit einer hohen Inzidenz zwischen dem 8. und 16. LT. Das Allgemeinbefinden und die Bewegungs- und Sauglust blieben dabei unbeeinflusst. Zum Zeitpunkt des ersten Östrus der Stute zeigten 59 % der Fohlen Diarrhoe. Unter den 41 %, die keinen Durchfall zum Zeitpunkt der Fohlenrosse hatten, waren Fohlen, die nie Durchfall vom 1. – 58. LT zeigten, aber auch welche die Diarrhoe entwickelten, als die Mutterstute sich nicht in Rosse befand. Ein Zusammenhang zwischen der Fohlenrosse der Stute und Durchfall bei ihrem Fohlen konnte nicht hergestellt werden. Es zeigte sich eine Tendenz, dass hohe Spiegel der Gesamt-IgG (>20 mg/ml) und IgG-anti-LPS von E. coli J5 (>120 RE/ml) nach der Kolostrumaufnahme im Zusammenhang mit einer geringeren Anzahl von Durchfalltagen innerhalb der ersten zwei Lebensmonate standen. C. perfringens und Enterobakterien waren gleichermaßen nachweisbar bei Fohlen mit Durchfall als auch bei unauffälligen Fohlen. Aus der Supplementierung von B. cereus var. toyoi ergab sich kein Effekt auf die Kotflora der Fohlen, außer auf die Gesamtkeimzahlen (GKZ) der aeroben Bakterien. Bei den Aerobiern im Fohlenkot konnte ein signifikanter Behandlungseffekt (p=0,012) festgestellt werden. Im ersten Milchkot der Fohlen waren GKZ von 4,5 x 104 KbE/g (200 mg Toyocerin-Gruppe) bis 5,0 x 105 KbE/g (50 mg Toyocerin-Gruppe) bei den aeroben Bakterien und GKZ von 2,4 x 105 KbE/g (200 mg Toyocerin-Gruppe) bis 4,7 x 106 KbE/g (50 mg Toyocerin-Gruppe) median bei den Anaerobiern nachweisbar. Danach stieg der Gehalt der aeroben und anaeroben Bakterien weiter bis zum 3. LT und stagnierte bis zum 16. LT. Während dieser Stagnationsphase trat bei 92 % der Fohlen (23/25) eine Veränderung der Kotkonsistenz bis hin zu Durchfällen auf. Vom 16. bis zum 58. LT sanken die Gehalte moderat bei den Aerobiern median am 58. LT auf 2,7 x 105 KbE/g (Placebo-Gruppe) bis 2,2 x 106 KbE/g (50 mg Toyocerin-Gruppe) und bei den Anaerobiern median am 58. LT auf 3,8 x 105 KbE/g (Placebo-Gruppe) bis 2,9 x 106 KbE/g (200 mg Toyocerin-Gruppe). Bis zum 58. LT näherte sich der Medianwert der aeroben und anaeroben Bakterien im Kot der Placebo-Gruppe dem Wert der Mutterstuten (gemessen am ersten Tag nach der Geburt) an. Innerhalb der ersten Lebenstage war eine hohe aerobe sowie anaerobe Keimzahl im Kot der Fohlen nachzuweisen, die sich oberhalb der Keimzahlen befand, die im Kot der Mutterstuten zum Zeitpunkt der Geburt gemessen wurde. Im Rahmen der Entwicklung und Etablierung der bakteriellen intestinalen Mikroflora wurde das Fohlenrossedurchfallgeschehen bei den Fohlen beobachtet. B. cereus var. toyoi hatte dabei keinen Einfluss auf die Anzahl der Fohlen mit Durchfall und den Gesundheitsstatus der Fohlen. / Diarrhoea is probably one of the most common problems in equine neonates. Almost all foals develop transient diarrhoea within the first weeks of life. Different viral, bacterial and parasitic causes are discussed. Between the 5th and the 15th day of the foal’s life, when their dam’s first post partum (p.p.) oestrus is expected, diarrhoea in foals is observed quite often. That is why it’s called “foal heat diarrhoea”. In literature establishment of intestinal microflora and maturation of the intestinal mucosa is responsible for the occurrence of diarrhoea in this period of life. But little is known about the development of the intestinal microflora in foals. Many probiotics are authorised as gut flora stabilisers in animal nutrition. Some studies proved positive effects of Bacillus (B.) cereus var. toyoi (Toyocerin®) on intestinal health in other species e.g. calves, piglets, broiler chicken, poultry and growing rabbits. The present study deals with the question if a supplementation of B. cereus var. toyoi lead to a stabilisation of the developing intestinal microflora and therefore to a reduction of diarrhoea in foals. A total of 25 mares and foals of a thoroughbred stud were included into the study. Foals were born between February and May 2011. From birth, the foals were randomly assigned to three treatment groups: placebo group (10 ml isotonic saline solution, n=8), 50 mg Toyocerin group (5 x 108 cfu B. cereus var. toyoi solved in 10 ml isotonic saline solution, n=7) and 200 mg Toyocerin group (2 x 109 cfu B. cereus var. toyoi solved in 10 ml isotonic saline solution, n=10). Placebo- and treatment groups were orally supplemented once a day starting on the 1st through to the 58th day of life. Determination of heart and respiratory rate, body temperature, body weight was realised according to a standardised protocol. Within the first day of life, on day 9, 16, 23, 30, 44, 58 and on the first day of diarrhoea faecal samples has been taken from the rectum or by the use of a collection bag. Blood samples were taken via jugular venipuncture on day 1, 9, 16, 30, 58 and on the first day of diarrhoea. Culture-depend methods were used to analyse the bacterial microflora. Serum IgG was analysed by a competitive ELISA, IgG-anti-LPS from E. coli J5 and IgG-anti-PLC-from-C. perfringens-1a by an indirect ELISA. 88 % of the foals developed diarrhoea (placebo 7/8, 50 mg Toyocerin 5/7, 200 mg Toyocerin 10/10) with a high incidence between the 8th and the 16th day of the foal’s life. Meanwhile, foals remained bright and alert and continued to nurse. At the time point of the first p.p. oestrus in the mares, 59 % of their foals showed signs of diarrhoea. Within the remaining 41 % there are foals that had no diarrhoea but there are also foals which had diarrhoea when the mare had not been in heat. Neonatal diarrhoea in foals is not linked to p.p. oestrus in their mares. There was a tendency, that high serum-IgG (> 20 mg/ml) and IgG-anti-LPS from E. coli J5 (> 120 RE/ml) after colostrum uptake were associated with lower diarrhoea severity in the first 58 days of the foal’s life. C. perfringens and enterobacteria can be found equally in foals with diarrhoea and in foals which are not afflicted. B. cereus var. toyoi supplementation had no effect on faecal bacteria in foals, except on aerobic bacteria (p=0,012). In the first milk faeces aerobic bacteria were detected in median from 4,5 x 104 cfu/g (200 mg Toyocerin-group) to 5,0 x 105 cfu/g (50 mg Toyocerin-group) and anaerobic bacteria were detected in median from 2,4 x 105 cfu/g (200 mg Toyocerin-group) to 4,7 x 106 cfu/g (50 mg Toyocerin-group). Afterwards the counts increased towards the 3rd day of life and stayed on a high level till the 16th day of life. During this stagnation in 92 % of the foals a change in faecal consistency and diarrhoea was observed. Afterwards, from the 16th though to the 58th day of life, the bacteria counts in the faeces moderately decreased in median for the aerobic bacteria on the 58th day of life down to 3,8 x 105 cfu/g (placebo-group) till 2,9 x 106 cfu/g (200 mg Toyocerin-group). On the 58th day of life the counts of aerobic and anaerobic bacteria in the faeces of the placebo-group approached the counts in the faeces of the mare (measured at the time point of birth). In the first days of foals’ life detection of aerobic and anaerobic bacteria in the faeces were high, and above the level of the bacteria counts in the faeces of the mare at the time point of birth. Foal heat diarrhoea is observed as a part of the development and establishment of bacterial intestinal microflora. B. cereus var. toyoi had no effect on the percentage of foals with diarrhoea and health status in the foals at that point.