Streamlining sporozoite isolation from mosquitoes by leveraging the dynamics of migration to the salivary glands

Background Sporozoites isolated from the salivary glands of Plasmodium-infected mosquitoes are a prerequisite for several basic and pre-clinical applications. Although salivary glands are pooled to maximize sporozoite recovery, insufficient yields pose logistical and analytical hurdles; thus, predicting yields prior to isolation would be valuable. Preceding oocyst densities in the midgut is an obvious candidate. However, it is unclear whether current understanding of its relationship with sporozoite densities can be used to maximize yields, or whether it can capture the potential density-dependence in rates of sporozoite invasion of the salivary glands. Methods This study presents a retrospective analysis of Anopheles stephensi mosquitoes infected with two strains of the rodent-specific Plasmodium berghei. Mean oocyst densities were estimated in the midguts earlier in the infection (11-15 days post-blood meal), with sporozoites pooled from the salivary glands later in the infection (17-29 days). Generalized linear mixed effects models were used to determine if (1) mean oocyst densities can predict sporozoite yields from pooled salivary glands, (2) whether these densities can capture differences in rates of sporozoite invasion of salivary glands, and (3), if the interaction between oocyst densities and time could be leveraged to boost overall yields. Results The non-linear effect of mean oocyst densities confirmed the role of density-dependent constraints in limiting yields beyond certain oocyst densities. Irrespective of oocyst densities however, the continued invasion of salivary glands by the sporozoites boosted recoveries over time (17-29 days post-blood meal) for either parasite strain. Conclusions Sporozoite invasion of the salivary glands over time can be leveraged to maximize yields for P. berghei. In general, however, invasion of the salivary glands over time is a critical fitness determinant for all Plasmodium species (extrinsic incubation period, EIP). Thus, delaying sporozoite collection could, in principle, substantially reduce dissection effort for any parasite within the genus, with the results also alluding to the potential for changes in sporozoites densities over time to modify infectivity for the next host.Host-parasite interactio

Digestibility and pricing of Chlorella sorokiniana meal for use in tilapia feeds

Several microalgae contain in excess of 50 % crude protein with amino acid profile comparable to that of fish meal. In addition, high polyunsaturated fatty acid contents encourage their use in animal feeding and nutrition, particularly in the formulation and processing of aquafeeds. This study aims at estimating the feasibility of Chlorella meal as feed ingredient for the feeding and nutrition of farmed tilapia based upon digestibility data. Juvenile tilapia were stocked in conical-bottomed tanks (200 L) with superficial, continuous water flow, and fed to apparent satiation in three daily meals with a reference diet and a test diet containing 30 % lyophilized Chlorella sorokiniana added of an inert marker. Feces were collected overnight by sedimentation in refrigerated, plastic containers coupled to the tanks and analyzed for determination of chemical composition and inert marker contents to estimate apparent digestibility coefficients (ADCs) of protein and energy of Chlorella meal; registered ADCs of Chlorella meal were 90.5 and 84.22, respectively. A pricing model considering the quantity of digestible nutrient was proposed based on ADCs of Chlorella and compared with the price of fishmeal (FM) and soybean meal (SBM). The indicative prices to elicit the use of Chlorella as a protein source rather than FM or SBM for the feed and nutrition of tilapia were 2.65 USD kg−1 and 0.66 USD kg−1, respectively