Comparison of the survival on ice of thawed Theileria parva sporozoites of different stocks cryoprotected by glycerol or sucrose.

Stabilates of Theileria parva sporozoites are mostly delivered in liquid nitrogen tanks to the East Coast fever immunization points. Using an in vitro titration model, we assessed the loss of infectivity of several stabilates when they are stored in ice baths for up to 24 h. Comparisons, with respect to rates of loss of infectivity, were made between T. parva stocks (Chitongo and Katete), cryoprotectants (sucrose and glycerol) and method of assessment (in vivo and in vitro techniques). Chitongo and Katete stabilates showed similar loss dynamics. The losses were 1-4% (depending on parasite stock) and 3% per hour of storage for glycerol and sucrose stabilates respectively, and the loss rates were not significantly different. The results suggest that Chitongo stabilates and sucrose cryoprotected suspensions can be delivered on ice as is done for Katete. A graphical relationship of in vitro effective dose at 50% infectivity (ED50) and in vivo protection rate was made. The relationship showed a 35% loss of protection for a relatively low corresponding increase of ED50 from 0.006 to 0.007 tick equivalent.

Infectivity of Theileria parva sporozoites following cryopreservation in four suspension media and multiple refreezing: evaluation by in vitro titration.

Theileria parva sporozoite stabilates are used for immunizing cattle against East Coast fever and in in vitro sporozoite neutralization assays. In this study, we attempted to identify a cheaper freezing medium and quantified the infectivity loss of sporozoites due to refreezing of stabilates, using an in vitro technique. Pools of stabilates prepared using Minimum Essential Medium (MEM), Roswell Park Memorial Institute (RPMI 1640), foetal calf serum (FCS) and phosphate-buffered saline (PBS) were compared. All were supplemented with bovine serum albumin except the FCS. RPMI 1640 was as effective as MEM in maintaining sporozoite infectivity while the infectivity in PBS and FCS reached only 59% and 67%, respectively. In a second experiment, a stabiiate based on MEM was subjected to several freeze-thaw cycles including various holding times on ice between thawing and refreezing. Refrozen stabilate gave an average sporozoite infectivity loss of 35% per cycle. The results indicate that RPMI can be used as a cheaper freezing medium for T. parva stabilates and that refrozen stabilate doses need to be adjusted for the 35% loss of infectivity.

Comparison of manual and homogenizer methods for preparation of tick-derived stabilates of Theileria parva: equivalence testing using an in vitro titration model.

Theileria parva sporozoite stabilates are used in the infection and treatment method of immunization, a widely accepted control option for East Coast fever in cattle. T. parva sporozoites are extracted from infected adult Rhipicephalus appendiculatus ticks either manually, using a pestle and a mortar, or by use of an electric homogenizer. A comparison of the two methods as a function of stabilate infectivity has never been documented. This study was designed to provide a quantitative comparison of stabilates produced by the two methods. The approach was to prepare batches of stabilate by both methods and then subject them to in vitro titration. Equivalence testing was then performed on the average effective doses (ED). The ratio of infective sporozoites yielded by the two methods was found to be 1.14 in favour of the manually ground stabilate with an upper limit of the 95% confidence interval equal to 1.3. We conclude that the choice of method rests more on costs, available infrastructure and standardization than on which method produces a richer sporozoite stabilate.

Theileria parva seroprevalence in traditionally kept cattle in southern Zambia and El Nino.

Sero-epidemiological surveys involving 27,526 cattle over a period of 8 years show that Theileria parva, the parasite causing East Coast fever (ECF) is found throughout southern Zambia. Higher values of T. parva sero-prevalence were observed in the plateau districts of Monze, Choma and Mazabuka than in the valley districts of Siavonga and Sinazongwe. Our results reveal a strong association between high T. parva sero-prevalence and the presence of the periodic climatic phenomenon known as the El Nino Southern Oscillation. More T. parva sero-positive samples were recorded during El Nino years (1997/98) (P<0.001) than other years in the study period. From this association, we conclude that Multiple El Nino Southern Oscillation Indices can be used to predict years with high or low ECF infection prevalence thereby contributing to the improved control of ECF in the area.