Transition models to assess risk factors for new and persistent trypanosome infections in cattle-analysis of longitudinal data from the Ghibe Valley, southwest Ethiopia.

The objective of this study was to apply transition models to distinguish between factors associated with both incident and persistent trypanosome infections. Data collected from 1561 cattle were analyzed from a long-term study involving 8 herds in which both trypanosome infections (a total of 56,931 cattle sampling-months) and tsetse (Glossina spp.) challenge were monitored monthly from March 1986 to March 1998. Both pour-on and insecticide-target tsetse control programs and mass treatment with diminazene aceturate before tsetse control were associated with significant decreases in both incidence and persistence of trypanosome infection relative to noncontrol periods, as were seasonal and sex effects. The magnitudes of the effects were, however, often different for new and persistent infections. For persistence of infection, there were 2 trends. In general, the duration of infection increased during the study, despite the regular treatment with diminazene aceturate. The transition model had 2 major benefits. The first was to identify an increasing duration of infections with time, taking into account other factors associated with increasing infection risk. The second was to highlight different patterns in the effects of certain factors on new and persistent trypanosome infections.

The impact of tick control on the productivity of indigenous cattle under ranch conditions in Uganda.

The impact of tick control on the productivity of indigenous cattle was investigated in three herds of East African shorthorn Zebu and Zebu x Ankole (Nganda) breeds, maintained under three tick management systems practised under ranch conditions: twice-a-week dipping, once-a-month dipping, and no tick control. The milk production and the growth rates of pre-weaned and post-weaned calves were monitored once a month over a period of 34 months. Milk offtake was 23% higher during the heavy rainy seasons than in the long dry seasons. Twice-a-week dipping increased the milk offtake by 21% in the second year of study and prolonged the duration of lactation in cows. Similarly, twice-a-week dipping increased the pre-weaning growth rate by 39% in the second year of study, but had no significant effect on the post-weaning growth rate. Generally, the growth rate of calves were greatest during the heavy rainy seasons and least during the long dry seasons. These results provide a basis for assessing the losses under different tick management systems in the various production systems.

Seroprevalences of vector-transmitted infections of small-holder dairy cattle in coastal Kenya.

A cross-sectional study was carried out from July to September 1989 in Kaloleni Division, Coast Province, Kenya to estimate the prevalence of vector-transmitted diseases in small-holder dairy cattle and to identify the risk factors associated with different management systems. One hundred and thirty of the 157 herds with dairy cattle in Kaloleni Division were surveyed. These were from three agro-ecological zones (coconut-cassava, cashew nut-cassava and livestock-millet), comprised two management systems (stall-feeding and herded grazing) and were herds with either dairy cattle only or with Zebu and dairy cattle. A formal questionnaire sought answers to questions on cattle health and management practices. A total of 734 dairy and 205 Zebu cattle in 78 dairy and 52 mixed (dairy and Zebu) herds were sampled and screened for haemoparasites (Trypanosoma, Anaplasma, Babesia, and Theileria infections). Sera were tested for antibodies to Theileria parva, using the schizonts-antigen indirect fluorescent-antibody (IFA) test and to antibodies for Babesia bigemina and antigens to Anaplasma marginale by enzyme-linked immunosorbent assay (ELISA). Packed-cell volume (PCV) also was measured. Tick-control measures were practised by all except three of the farmers. Despite this, overall seroprevalence to T. parva was >70%--suggesting either that control practices were not strictly implemented or they were ineffective. The seroprevalence of T. parva in adult cattle kept in stall-feeding systems in the coconut-cassava zone was significantly lower (57+/-8% (S.E.)) than in herded-grazing systems (79+/-3%) and there was no association between antibody prevalence and age of cattle in this zone. Antibody prevalences in cattle in the cashew nut-cassava and the drier livestock-millet zone increased with age. Cattle in herded-grazing systems had an overall lower seroprevalence of T. parva infection in the livestock-millet zone (45+/-6%) than in the other two zones. Analysis was confined to the coconut-cassava zone for B. bigemina and to the coconut-cassava and cashew nut-cassava zones for A. marginale. Mean prevalences of B. bigemina were 40.9+/-9 and 73+/-6% for dairy cattle under stall-feeding and herded-grazing systems, respectively, and increased with age. Antigen prevalences of A. marginale were over 80% in all age groups of cattle in the coconut-cassava and cashew nut-cassava zones. Overall trypanosome prevalence in cattle was <1%. Trypanocidal treatment was uncommon. The variations in antibody prevalence associated with risk factors such as feeding system, agro-ecological zone and age of animal suggest that management system influenced exposure to tick-borne infection (particularly, T. parva infections) in small-holder dairy cattle in coastal Kenya.

A longitudinal study of disease incidence and case-fatality risks on small-holder dairy farms in coastal Kenya.

A longitudinal study was carried out in the coastal lowlands coconut-cassava agro-ecological zone of Kaloleni Division, Coast Province, Kenya between June 1990 and December 1991 to estimate disease incidence and cause-specific case-fatality risk in an average of 120 cattle in 26 small-holder dairy herds kept in two grazing-management systems. East Coast fever (ECF) was the predominant disease diagnosed; the mean monthly incidence rate was 2.5 and 6.9% in animals < or = 18 months of age under stall-fed and herded-grazing systems, respectively. In cattle > 18 months of age, the monthly incidence rate was < 1%. The 6-month ECF incidence rate was 20+/-8% (S.E.) in the stall-feeding system compared with 39+/-7% in the herded-grazing systems. There was a gradual increase in antibody prevalence with age to over 90% in cattle over 18 months of age in herded-grazing systems, whilst less than a third of cattle in the stall-feeding systems were sero-positive at any age. Overall accumulated mortality to 18 months of age was estimated to be 56%. Annual mortality in cattle > 18 months averaged 9%. Cattle managed in the herded-grazing system had a 60% higher mortality, although not significantly so, than those fed in stalls. Deaths due to ECF accounted for over two-thirds of the deaths. ECF was then the major disease constraint to small-holder dairy production in the coconut zone of coastal Kenya. Clinical cases occur the whole year round (especially in young stock)--despite apparent tick control, and in both herded-grazing and stall-feeding system.

Identification of a target population for immunisation against East Coast fever in coastal Kenya.

Two experiments were carried out to identify the target population of cattle for immunisation against East Coast fever (ECF) using the infection-and-treatment method. Firstly, a sentinel-calf study was used to determine the age window for ECF immunisation by determining ages at clinical detection of infection with Theileria parva. Six groups of five naive cross-bred (Bos taurus/Bos indicus) male calves, introduced at intervals of 2 months at a mean age of 26 days, were exposed to natural tick challenge on a high ECF-risk, small-holder farm in the coastal lowland, coconut-cassava agro-ecological zone of coastal Kenya. Secondly, a challenge study evaluated the relationship between the presence of T. parva antibodies and immunity. Ten indigenous adult Zebu cattle and nine Zebu young stock purchased from farmers in the same zone, and eight cross-bred calves (survivors of the sentinel-calf study) were challenged with 10 times the immunising dose of T. parva Marikebuni stock. Twenty-four of these 27 cattle had high antibody titres before challenge. Two cross-bred calves, obtained from an ECF-free area and seronegative to T. parva schizont antigen, also were challenged and used as susceptible controls. Twenty-five (83%) of the 30 sentinel calves contracted ECF over an age range of 36-116 days (mean 72 days). The remaining five calves died of other causes within 2 months of arrival on the farm. Fourteen of the 25 calves survived the infection and developed antibodies to T. parva. Despite tick control, seven of these 14 calves had a second episode of ECF and two died. In total, 13 of the 25 calves that contracted ECF died. Only one of 19 indigenous Zebu animals developed clinical ECF when challenged with T. parva Marikebuni (mild clinical signs with spontaneous recovery). Of the eight cross-bred survivors from the first experiment, only one succumbed to ECF when challenged and it died. Both susceptible cross-bred calves developed severe clinical signs of ECF and one died. The experimental studies show that in the high ECF-risk areas of the coconut-cassava zone of coastal Kenya, immunisation against ECF in cross-bred (B. taurus/B. indicus) cattle should be targeted at an early age (preferably within 1-2 months of birth).

The incidence of new and the prevalence and persistence of recurrent trypanosome infections in cattle in southwest Ethiopia exposed to a high challenge with drug-resistant parasites.

A method is described for calculating new infection incidence from monthly field data collected between April 1994 and February 1998 from an average of 770 Ethiopian Highland Zebu cattle maintained under traditional management in the Ghibe valley, southwest Ethiopia and exposed to a high challenge with drug-resistant parasites. Each month cattle with a packed cell volume (PCV)<26% and detected as parasitaemic, or cattle showing clinical signs of trypanosomosis, were treated with diminazene aceturate at 3.5 mg/kg body weight. An infection was defined as a new infection if it was preceded by 2 previous months in which both samples had a PCV>/=26% and were not detected with trypanosomes. Using this definition the average monthly incidence of infections of Trypanosoma congolense in cattle over 36 months of age was 13.3%, and the prevalence of recurrent infections 13.0%. Assuming that an animal had recovered from infection when PCV again returned to 26% without parasites being detected, mean persistence of infection was 3.8 months (median between 2 and 3 months). In contrast, T. vivax infections were susceptible to diminazene. The incidence of T. vivax infections in adults was lower than for T. congolense (2.8%), and they were less pathogenic (mean reduction of 3.1% units of PCV due to T. vivax infection compared with 4.6% units for T. congolense). Also, fewer cases were treated. Calves were first detected parasitaemic at an average age of 8.8 months and their infections persisted longer than those in adults. The effect of age on incidence of infection was not significant beyond 15 months of age. Adult male cattle appeared to be more susceptible to T. congolense infection than adult female cattle.

Use of deltamethrin 'pour-on' insecticide for the control of cattle trypanosomosis in the presence of high tsetse invasion.

A deltamethrin 'pour-on' insecticide was applied monthly to over 2000 cattle exposed to a high challenge of drug-resistant trypanosomes and high tsetse re-invasion pressure in the Ghibe valley, south-west Ethiopia. Blood samples were taken monthly from an average of 760 cattle for determination of PCV and presence of trypanosomes. The area of the valley is approximately 350 km2 and the cattle grazed in roughly four locations covering about a quarter to half of the area. Two years before the trial commenced, Glossina morsitans submorsitans Newstead (Diptera: Glossinidae) began to invade the valley. Despite the use of the pour-on the mean apparent density of G. m. submorsitans continued to rise, and, during the 4 years of tsetse control, was more than three-fold higher than that recorded during the previous 18 months. Over the same period there was little change in the apparent density of Glossina pallidipes Austen (Diptera: Glossinidae). By contrast, the mean monthly prevalence of trypanosome infections in cattle over 36 months of age decreased from 38.3 to 29.0%, the incidence of new infections decreased from 26.6 to 16.0% (a reduction of 40%), and packed cell volume in cattle increased from 21.7 to 24.1%. Evidence of a change in apparent parasite transmission rate was demonstrated by regression of infection incidence in cattle on the logarithm of apparent density of G. m. submorsitans. Before the trial started the regression coefficient was 45.8 +/- 6.3 and this reduced to 9.2 +/- 2.5% incidence per log(e) (flies/trap/day) during the period of tsetse control. It was concluded that this indicated reductions in tsetse numbers in the immediate vicinities of cattle in a way that was not reflected in overall tsetse catches. Nevertheless, the comparatively high levels of trypanosome prevalence that persisted in the cattle demonstrates that, where invasion prevalence is high, treatment of small pockets of cattle will not eradicate tsetse. To achieve more significant reduction in trypanosome prevalence in cattle, integrated methods of control utilizing target barriers in the major routes of invasion will be needed.

The relationship between the parasitological prevalence of trypanosomal infections in cattle and herd average packed cell volume.

The relationship between prevalence of trypanosomal infections (Trypanosoma congolense) and average packed cell volume (PCV) in herds of communally managed adult Angoni breed cattle was investigated in four districts of eastern Zambia. In all areas, regression analyses showed that the herd average PCV decreased with increasing prevalence of trypanosomal infections. The slope of the equation between average PCV and trypanosome prevalence decreased with increasing prevalence of trypanosomal infections. For the same increase in prevalence of trypanosomal infection, the decrease in herd PCV was higher in the areas with low to medium prevalence. Season of sampling also determined the slope of the regression equation. For the same increase in prevalence of trypanosomal infection, the decrease in herd PCV was higher during the dry compared to the rainy season suggesting that trypanosomosis is less well tolerated during the dry season. Results from the study suggest that the relationship between the prevalence of trypanosomal infections and herd average PCV could be a useful tool in the management of trypanosomosis and planning of its control. Reasons for the spatial and temporal variations in the relationship are discussed.

A statistically derived index for classifying East Coast fever reactions in cattle challenged with Theileria parva under experimental conditions.

A statistically derived disease reaction index based on parasitological, clinical and haematological measurements observed in 309 5 to 8-month-old Boran cattle following laboratory challenge with Theileria parva is described. Principal component analysis was applied to 13 measures including first appearance of schizonts, first appearance of piroplasms and first occurrence of pyrexia, together with the duration and severity of these symptoms, and white blood cell count. The first principal component, which was based on approximately equal contributions of the 13 variables, provided the definition for the disease reaction index, defined on a scale of 0-10. As well as providing a more objective measure of the severity of the reaction, the continuous nature of the index score enables more powerful statistical analysis of the data compared with that which has been previously possible through clinically derived categories of non-, mild, moderate and severe reactions.

Estimating the effects of tsetse control on livestock productivity--a case study in southwest Ethiopia.

A tsetse control campaign was started in January 1991 using a synthetic pyrethroid cypermethrin 'pour-on' applied monthly to cattle exposed to high levels of drug-resistant trypanosomes in the Ghibe valley of southwest Ethiopia. In December 1992, a cost-recovery scheme was introduced, and thereafter farmers paid for treatment. An average of 100 village Ethiopian Highland Zebu cattle were monitored monthly from March 1986 to February 1997. Individual animals in this herd were treated with diminazene aceturate (Berenil, Hoechst, Germany) at 3.5 mg/kg when trypanosomes were detected and their packed cell volume was less than 26%. Superimposed on this systematic trypanocidal chemotherapy, control of tsetse resulted in average reductions from 1992 to 1996 of 95% and 75% in the mean relative densities of tsetse and biting flies, respectively, and of 63% in the prevalence of trypanosomal infections in cattle. Despite these reductions, there was no significant increase in the body weight of the cows, calving rate or the mean body weight of calves at 12 months of age. There was, however, an average decrease of 57% in calf mortality (including still births) by 12 months of age, an increase of 49% in the ratio of live calves under 12 months of age to cows over 36 months of age, and an increase of 8% in the body weight of adult males.

Population dynamics of ticks on indigenous cattle in a pastoral dry to semi-arid rangeland zone of Uganda.

Studies on seasonality and population dynamics of ticks on indigenous cattle and their crosses (calves) were carried out in Buruli Ranching Scheme, Nabiswera, Luwero district of Uganda on three treatment groups of animals: group 1 (twice a week dipping), group 2 (once a month dipping) and group 3 (no tick control). During this study, four major species of ticks of economic importance were recorded in decreasing order of abundance: Rhipicephalus appendiculatus, Rhipicephalus evertsi evertsi, Amblyomma variegatum and Boophilus decoloratus. Of these ticks, a seasonal pattern of activity was only observed in R. appendiculatus ticks, with peak activities occurring during rainy seasons. Greater numbers of ticks were recorded on cows than calves in the three treatment groups, with the exception of A. variegatum where the reverse occurred. The mean numbers of ticks per animal were highly significantly different (p < 0.01) when group 1 animals were compared with group 2 and 3 animals. However, no significant differences (p > 0.05) were observed in mean tick numbers between group 2 and 3 animals. Highly significant differences (p < 0.01) were observed in mean tick numbers on cows and calves (more than 12 months old) in different calving seasons. The state of lactation only affected tick counts on cows in group 1; significantly more ticks (p < 0.01) were observed in lactating than non-lactating cows. Furthermore, significantly greater (p < 0.05) numbers of ticks (with the exception of B. decoloratus) were recorded during the second year of study (March 1992-May 1993) than the first year (January 1991-February 1992), despite lower rainfall during the former period.

Power evaluation of small drug and vaccine experiments with binary outcomes.

Power analysis based on the Fisher exact test is used to evaluate the usefulness of alternative designs for small drug and vaccine experiments with a binary, success or failure, outcome. The emphasis is on the comparison between designs with equal and unequal allocations of animals to treated and control groups having the same total sample size, and between designs with equal and unequal allocations but with one or two fewer animals in the control than the treated group. Designs with unequal allocation can sometimes be more powerful at a given level of statistical significance alpha than designs with equal allocation. Graphs and a table are presented to aid the choice of suitable designs for (1-beta) = 0.80 and alpha = 0.05.

Use of insecticide-impregnated targets for the control of tsetse flies (Glossina spp.) and trypanosomiasis occurring in cattle in an area of south-west Ethiopia with a high prevalence of drug-resistant trypanosomes.

In the Ghibe valley, south-west Ethiopia, a tsetse control trial using deltamethrin-impregnated targets was started in May 1990. The mean relative density of the main vector, Glossina pallidipes, fell from 2.1 flies per trap per day in the 12 months prior to introduction of tsetse control to 0.41 flies per trap per day in the 12 months after tsetse control was initiated. The annual mean prevalence of Trypanosoma congolense infections in cattle fell from 32% in the 2 years before tsetse control to 13% in the 2 years following deployment of targets. The largest reduction occurred in the first quarter of 1991 when the mean monthly trypanosome prevalence was 5%. There was also a decrease of approximately 75% in the estimated rate of relapse of trypanosome infections in cattle after treatment with diminazene aceturate. However, in June 1991, socio-political disturbances occurred in Ethiopia and were associated with substantial thefts of targets. Following these thefts, the mean relative density of G. pallidipes rose from 0.41 flies per trap per day in the period May 1990-April 1991, to 1.29 flies per trap per day in the period April 1992-March 1993. Associated with this rise, the mean trypanosome prevalence in cattle increased from 13% in the period May 1991-April 1992 to 28% in the period May 1992-April 1993. This was similar to the mean trypanosome prevalence in cattle during the pre-control period. Thus, while effective tsetse control methods can be used to reduce the transmission of trypanosomes their long-term impact is dependent on their sustainability.

Effect of tsetse control on trypanosome prevalence in livestock: problems of experimental design and statistical interpretation--a case study in northern Côte d'Ivoire.

As part of a study on livestock productivity under trypanosomosis risk in the region of Boundiali, northern Côte d'Ivoire, 21 herds of cattle (N'Dama, Baoulé and Zebu crosses) and 20 flocks of Djallonké and Djallonké x Sahel sheep were monitored monthly for body weight, packed red cell volume and trypanosomal parasitaemia over various periods between January 1984 and December 1992. A tsetse control campaign using biconical traps impregnated with alpha-cypermethrin started in December 1987. Tsetse control reduced the relative tsetse density by over 95% between 1988 and 1992, and this was associated with reductions in the prevalence of Trypanosoma congolense over the same period of over 90% both in sheep and cattle. Average reductions in the prevalence of T. vivax were lower, on average 68% in adults and 85% in young animals. Attempts were made in the design of the study to allow comparisons between controlled and uncontrolled areas; however, there were too many confounding and uncontrollable factors to allow such comparisons to be made. It was necessary, therefore, to compare data collected from all herds and flocks before and after the intervention, with the consequential difficulties in accounting for uncontrollable year-to-year variations in factors affecting trypanosome prevalence in livestock.

Variations in packed red cell volume and trypanosome prevalence and relationships with reproductive traits in east African Zebu cows exposed to drug-resistant trypanosomes.

Approximately 320 East African Zebu cows over 36 months of age were monitored monthly from 1986 to 1992 in nine village herds in an area of high trypanosomiasis risk in southwest Ethiopia where there was resistance to all available trypanocidal drugs. Cows were individually treated with diminazene aceturate, either when they were detected parasitaemic and their packed red cell volume (PCV) decreased below 26%, or when they showed clinical signs of trypanosomiasis. The average annual monthly trypanosome prevalence was 24% and the number of treatments of diminazene aceturate per cow per year was 3.1, both of which increased with age. Mean PCV decreased and mean trypanosome prevalence increased during lactation. There was a significant linear association between the time detected parasitaemic during the first 150 days of lactation and calving interval. When corrected for frequency of parasitaemia and treatment there was also an average reduction of 8.4 +/- 2.6 days in calving interval per % unit increase in PCV. Age at first calving decreased by 0.44 +/- 0.26 months per % unit increase in mean PCV maintained between 24 and 30 months corrected for parasitaemia and treatment. The percentage of pregnancies terminating in abortions significantly increased from 6.8 +/- 1.0% to 10.4 +/- 1.3% when cows detected parasitaemic at least once during the last three months of pregnancy were compared with cows not detected parasitaemic. The largest increase to 19.4 +/- 4.3% was in cows with low mean PCVs < 22%. It was concluded that cows which were able to maintain higher than average PCVs when parasitaemic showed superior reproductive performance than those with lower than average PCVs.

Epidemiology of bovine trypanosomiasis in the Ghibe valley, southwest Ethiopia. 1. Tsetse challenge and its relationship to trypanosome prevalence in cattle.

In this, the first of a series of papers on the epidemiology of bovine trypanosomiasis in the Ghibe valley, southwest Ethiopia, the tsetse populations and their relationships to the prevalence of trypanosome infections in cattle are described. The tsetse challenge to cattle at two sites sites in the area was estimated as the product of tsetse relative density and the trypanosome infection rate in flies. The proportion of feeds taken by tsetse from cattle was also considered. Three tsetse species were detected in the area, Glossina pallidipes, G. fuscipes and G. morsitans submorsitans. A significant correlation (r = 0.60, P < 0.001) was observed between the mean monthly estimates of tsetse challenge due to G. pallidipes and the prevalence of trypanosome infections in cattle the following month at one site, whilst at the other, no significant relationship was observed (P = 0.08). The tsetse density at both sites showed seasonal changes which were related to the monthly rainfall. Finally, variations in tsetse density appeared to be the main factor responsible for variation in tsetse challenge and thus trypanosome prevalence in cattle.

Epidemiology of bovine trypanosomiasis in the Ghibe valley, southwest Ethiopia. 2. Factors associated with variations in trypanosome prevalence, incidence of new infections and prevalence of recurrent infections.

An average of 840 East African Zebu cattle from nine herds in the Ghibe valley, southwest Ethiopia were monitored from January 1986 to April 1990. Each month blood samples were collected for analysis of packed red cell volume (PCV) and detection of trypanosomes. Animals found to be parasitaemic and with a PCV less than 26% were treated with diminazene aceturate at a dose of 3.5 mg/kg body weight. The majority of infections were associated with Trypanosoma congolense (84% of infections in adult cattle and 71% in cattle less than 24 months of age), and the mean percentage of adult animals detected parasitaemic 1 month after treatment of an infection with T. congolense was 27%. In order to assess possible existence of drug resistance, a model was applied which allowed monthly incidences of new infections to be distinguished from recurrent infections. This model showed that the monthly incidence of new infections of T. congolense in adult cattle increased significantly from 11% in 1986 to 24% in 1989 following a concomitant increase in the tsetse challenge. The corresponding increase in overall prevalence of T. congolense was from 17% to 38% and the mean prevalence of recurrent infections increased significantly from 6% to 14%. These findings ruled out the possibility that the high prevalence of trypanosome infections in cattle was due only to a high tsetse challenge and pointed to the existence of T. congolense populations which expressed resistance to diminazene. There were variations associated with season, herd, age and sex in the incidence of new infections, prevalence of recurrent infections and relapse to treatment.

Epidemiology of bovine trypanosomiasis in the Ghibe valley, southwest Ethiopia. 3. Occurrence of populations of Trypanosoma congolense resistant to diminazene, isometamidium and homidium.

In July 1989, blood samples were collected from parasitaemic cattle in the Ghibe valley, Ethiopia, frozen in liquid nitrogen and transported to Nairobi, Kenya. Twelve of the stabilates were inoculated into individual Boran (Bos indicus) calves and characterised for their sensitivity, in turn, to diminazene aceturate (Berenil), isometamidium chloride (Samorin) and homidium chloride (Novidium). All 12 stabilates produced infections which were shown to be Trypanosoma congolense and resistant to treatment with diminazene aceturate at a dose of 7.0 mg kg-1 body weight (b.w.). Eleven of the infections were also resistant to isometamidium chloride at a dose of 0.5 mg kg-1 b.w. and homidium chloride at a dose of 1.0 mg kg-1 b.w. The drug-sensitivity phenotypes of three of the same isolates were also determined in goats which were each treated with only one of the three trypanocides: all expressed the same phenotypes as the populations expressed in the aforementioned Boran calves. Five clones were derived from one of the isolates which expressed a high level of resistance to all three trypanocides; each clone expressed high levels of resistance to all three trypanocides when characterised in mice. Thus, the multi-resistance phenotype of the parental isolate was associated with expression of mutli-resistance by individual trypanosomes. Finally, molecular karyotypes and electrophoretic variants of six enzymes were determined for seven and eight of the isolates, respectively. Six different karyotypes were observed and all eight of the latter isolates belonged to different zymodemes, indicating that the multi-resistance phenotype at Ghibe was associated with many genetically distinct populations.

The effect of udder preparation before milking and contamination from the milking plant on bacterial numbers in bulk milk of eight dairy herds.

The effect of teat washing and drying on bacterial numbers in bulk milk was compared with that of no teat preparation in eight commercial herds over one year. Using in-line milk samplers, milk was collected at various points during its passage through the milking plant and the samples were used to establish the relative significance of the sources of contamination of raw milk. Teat washing and drying of cows housed during winter reduced the total counts by 40% and streptococcal and coliform counts by 50%. Bacterial counts were significantly lower in cows at pasture during the summer and there was no reduction in count due to teat washing and drying. Bacteriological counts increased at each stage as the milk passed through the milking machine. The milking equipment significantly increased the total colony count by between 2000 and 3000/ml, and the bulk tank added a further 1500 to 2000/ml. The mean rinse bacterial counts of the milking equipment were higher in summer than winter, averaging 4.4 X 10(7) bacteria/m2 compared with 3.5 X 10(7)/m2 respectively. Although this level of bacterial contamination of the equipment is high by current standards, very low bulk milk bacterial counts were nevertheless achieved, particularly in the summer. This confirms that organisms from this source are not a major contaminant of the bulk milk. There was a very poor correlation between rinse counts and the bulk milk bacterial count, but a strong correlation (0.98) between total and streptococcal counts of the bulk milk. The unreliability of the use of rinse techniques to assess the contribution of milking equipment to bacterial counts of raw milk is emphasized.

The association between lameness and fertility in dairy cows.

One thousand, four hundred and ninety-one lactations in 770 Friesian, Holstein and Ayrshire crossbred cows have been used to study the associations between lameness and fertility. Lameness was associated with a longer interval between calving and first service and a longer interval between calving and conception. The largest increases in these intervals, of 17 and 30 days, respectively, occurred in cows with either sole or white line lesions occurring between 36 and 70 days after calving. The conception rate during the 63 days before a diagnosis of lameness was made was lower (31 per cent) than at other times (40 per cent).