Estimation of free-roaming domestic dog population size: Investigation of three methods including an Unmanned Aerial Vehicle (UAV) based approach

Population size estimation is performed for several reasons including disease surveillance and control, for example to design adequate control strategies such as vaccination programs or to estimate a vaccination campaign coverage. In this study, we aimed at investigating the possibility of using Unmanned Aerial Vehicles (UAV) to estimate the size of free-roaming domestic dog (FRDD) populations and compare the results with two regularly used methods for population estimations: foot-patrol transect survey and the human: dog ratio estimation. Three studies sites of one square kilometer were selected in Petén department, Guatemala. A door-to-door survey was conducted in which all available dogs were marked with a collar and owner were interviewed. The day after, UAV flight were performed twice during two consecutive days per study site. The UAV's camera was set to regularly take pictures and cover the entire surface of the selected areas. Simultaneously to the UAV's flight, a foot-patrol transect survey was performed and the number of collared and non-collared dogs were recorded. Data collected during the interviews and the number of dogs counted during the foot-patrol transects informed a capture-recapture (CR) model fit into a Bayesian inferential framework to estimate the dog population size, which was found to be 78, 259, and 413 in the three study sites. The difference of the CR model estimates compared to previously available dog census count (110 and 289) can be explained by the fact that the study population addressed by the different methods differs. The human: dog ratio covered the same study population as the dog census and tended to underestimate the FRDD population size (97 and 161). Under the conditions within this study, the total number of dogs identified on the UAV pictures was 11, 96, and 71 for the three regions (compared to the total number of dogs counted during the foot-patrol transects of 112, 354 and 211). In addition, the quality of the UAV pictures was not sufficient to assess the presence of a mark on the spotted dogs. Therefore, no CR model could be implemented to estimate the size of the FRDD using UAV. We discussed ways for improving the use of UAV for this purpose, such as flying at a lower altitude in study area wisely chosen. We also suggest to investigate the possibility of using infrared camera and automatic detection of the dogs to increase visibility of the dogs in the pictures and limit workload of finding them. Finally, we discuss the need of using models, such as spatial capture-recapture models to obtain reliable estimates of the FRDD population. This publication may provide helpful directions to design dog population size estimation methods using UAV

Engineering Escherichia coli BL21(DE3) Derivative Strains To Minimize E. coli Protein Contamination after Purification by Immobilized Metal Affinity Chromatography ▿ † ‡

Recombinant His-tagged proteins expressed in Escherichia coli and purified by immobilized metal affinity chromatography (IMAC) are commonly coeluted with native E. coli proteins, especially if the recombinant protein is expressed at a low level. The E. coli contaminants display high affinity to divalent nickel or cobalt ions, mainly due to the presence of clustered histidine residues or biologically relevant metal binding sites. To improve the final purity of expressed His-tagged protein, we engineered E. coli BL21(DE3) expression strains in which the most recurring contaminants are either expressed with an alternative tag or mutated to decrease their affinity to divalent cations. The current study presents the design, engineering, and characterization of two E. coli BL21(DE3) derivatives, NiCo21(DE3) and NiCo22(DE3), which express the endogenous proteins SlyD, Can, ArnA, and (optionally) AceE fused at their C terminus to a chitin binding domain (CBD) and the protein GlmS, with six surface histidines replaced by alanines. We show that each E. coli CBD-tagged protein remains active and can be efficiently eliminated from an IMAC elution fraction using a chitin column flowthrough step, while the modification of GlmS results in loss of affinity for nickel-containing resin. The “NiCo” strains uniquely complement existing methods for improving the purity of recombinant His-tagged protein