Kinetics of Expression of Two Major Plasmodium berghei Antigens in the Mosquito Vector, Anopheles stephensi

ABSTRACT. Expression of a 21 kDa determinant (Pbs21), first detected on the surface of ookinetes, and of the circumsporozoite protein (CSP) was studied by immunofluorescence and Western blots during the developmental cycle of Plasmodium berghei in the mosquito A nopheles stephensi . The expression of Pbs21 was predominantly localised on the ookinete surface one day after the infectious blood meal, and thereafter reactivity declined to a minimum on days 2 and 3, the time of onset of oocyst development. A gradual increase in fluorescence was observed on the oocysts from day 6 that was retained until day 17 post-infection. In contrast, sporozoites released from oocysts or salivary glands showed little or no antibody labelling with anti-Pbs21. Circumsporozoite protein was not detectable in any rnidgut preparations until 5–6 days after feeding, when reactivity was observed against immature oocysts. Expression then continued and increased throughout oocyst and sporozoite development. Western blots confirmed that Pbs21 was expressed minimally during the oocyst development but was not detectable in sporozoites. Co-localisation of anti-Pbs21 and anti-CSP monoclonal antibodies to the 50 kDa and 60 kDa bands in Western blots of sporozoite suggests immunological cross-reactivity between the CSP and the anti-21 kDa antibodies.     

Isolation and characterization of microsatellites in Chinese soft‐shelled turtle,Pelodiscus sinensis

Fifteen polymorphic microsatellite loci were developed for the Chinese soft‐shelled turtle (Pelodiscus sinensis) from the (GT)_n microsatellite‐enriched genomic library, using the fast isolation by amplified fragment length polymorphism of sequences containing repeats protocol. The polymorphism of all 15 loci ranged from two to seven alleles with observed heterozygosities ranging from 0.03 to 0.98 (mean 0.43) in one population of 40 individuals. These novel loci will be helpful for understanding the population structure at genetic level and marker‐assisted breeding of this vulnerable species.     

Isolation and characterization of microsatellite markers from Sarcocystis neurona,a causative agent of equine protozoal myeloencephalitis

The population genetics and systematics of coccidian parasites of the genus Sarcocystis remain poorly defined, notwithstanding their relevency to veterinary and human health. Despite opportunities for sexual recombination, nonrecombinant parasite clones characterized by distinct transmission and pathogenesis traits persist in related parasites (i.e. Toxoplasma gondii). In order to determine whether this may be generally true for parasitic coccidia, and to address evolutionary and taxonomic problems within the genus Sarcocystis, we isolated 12 polymorphic microsatellite markers (four to 14 alleles) for Sarcocystis neurona, the major causative agent of equine protozoal myeloencephalitis (EPM).