Introduction: Knowledge of the geographical distribution of Leishmania species allows guiding the sampling to little-studied areas and implementing strategies to define risk zones and priority areas for control.Objective: Given that there is no publication that collects this information, the search, review, and compilation of the available scientific literature that has identified species in Colombia is presented in this paper.Materials and methods: A bibliographic search was performed in PubMed, Web of Knowledge, Google Scholar, SciELO and LILACS with the terms “(Leishmania OR Leishmaniasis) AND species AND Colombia”, without restrictions on publication year, language or infected organism; records of national scientific events and repositories of theses from Colombian universities were also included.Results: Eighty-six scientific documents published between 1985 and 2017 were found in which the species of Leishmania and their geographical origin were indicated. The species reported, in descending order of frequency, were: Leishmania (Viannia) panamensis, L. (V.) braziliensis, L. (V.) guyanensis, L. (Leishmania) infantum, L. (L.) amazonensis, L. (L.) mexicana, L. (V.) colombiensis, L. (V.) lainsoni and L. (V.) equatorensis; the last three were found with the same frequency. Leishmania species were reported from 29 departments. Conclusion: Information on the distribution of Leishmania species in Colombia is limited; therefore, it is necessary to gather existing data and propose studies that consolidate the distribution maps of Leishmania species in Colombia. This would allow the detection of areas where species have not been identified as well as the comparison of existing parasite and vector distributions.

In Colombia, nine species of parasites of the genus Leishmania circulate in more than 20 sand fly species, putting at risk of contracting the disease approximately 60% of the population. The Federico Lleras Acosta Dermatological Center, a reference center in Colombia, has been treating patients with cutaneous and mucosal leishmaniasis for more than 15 years, identifying the infecting Leishmania species from different clinical samples, and recording systematically all the epidemiological and geographic information related to each diagnosed patient. With this valuable information, the objective of this work was to perform a long term and large-scale study, aiming to identify the Leishmania species circulating in Colombia from clinical samples from 1999 to 2016, and to assess their current and potential spatial distribution. In all, four Leishmania species were identified in 688 samples from 183 municipalities distributed in 28 of the 32 departments of the country, and 387 records were georeferenced, from 20 departments. The most widespread species was L. (V.) braziliensis, showing new collection records, and the species related to areas with highest leishmaniasis transmission was L. (V.) panamensis. Ecological niche models were built for the three species that had more than 20 georeferenced records, showing a potential distribution for L. (V.) braziliensis on 42% of the national territory mainly in the interandean valleys, and the Orinoquia and Amazon regions. Leishmania (V.) guyanensis potential distribution covers 36% of Colombia continental territory with a spatial distribution similar to that of L. (V.) braziliensis. There was a marked tendency of L. (V.) panamensis to be distributed in the northwest of the country occupying 35% of the national area and mainly in areas of transformed ecosystems. Species were identified in patients from areas where the occurrence of cases was unprecedented, which suggests that the distribution of Leishmania may be greater than currently known. To improve the predictive capacity of the models, we suggest incorporating, in future studies, Leishmania samples from vectors and reservoirs that have a greater dependence on environmental variables. Our results are an important tool for health systems because they allow potential areas of transmission and information gaps to be identified.

Mitochondria are believed to play an important role in shaping the intracellular Ca²⁺ transients during skeletal muscle contraction. There is discussion about whether mitochondrial matrix Ca²⁺ dynamics always mirror the cytoplasmic changes and whether this happens in vivo in whole organisms. In this study, we characterized cytosolic and mitochondrial Ca²⁺ signals during spontaneous skeletal muscle contractions in zebrafish embryos expressing bioluminescent GFP-aequorin (GA, cytoplasm) and mitoGFP-aequorin (mitoGA, trapped in the mitochondrial matrix). The Ca²⁺ transients measured with GA and mitoGA reflected contractions of the trunk observed by transmitted light. The mitochondrial uncoupler FCCP and the inhibitor of the mitochondrial calcium uniporter (MCU), DS16570511, abolished mitochondrial Ca²⁺ transients whereas they increased the frequency of cytosolic Ca²⁺ transients and muscle contractions, confirming the subcellular localization of mitoGA. Mitochondrial Ca²⁺ dynamics were also determined with mitoGA and were found to follow closely cytoplasmic changes, with a slower decay. Cytoplasmic Ca²⁺ kinetics and propagation along the trunk and tail were characterized with GA and with the genetically encoded fluorescent Ca²⁺ indicator, Twitch-4. Although fluorescence provided a better spatio-temporal resolution, GA was able to resolve the same kinetic parameters while allowing continuous measurements for hours.

Zebrafish embryos have been proposed as a cost-effective vertebrate model to study heart function. Many fluorescent genetically encoded Ca²⁺ indicators (GECIs) have been developed, but those with ratiometric readout seem more appropriate to image a moving organ such as the heart. Four ratiometric GECIs based on troponin C, TN-XXL, Twitch-1, Twitch-2B, and Twitch-4 were expressed transiently in the heart of zebrafish embryos. Their emission ratio reported the Ca²⁺ levels in both the atrium and the ventricle. We measured several kinetic parameters of the Ca²⁺ transients: systolic and diastolic ratio, the amplitude of the systolic Ca²⁺ rise, the heart rate, as well as the rise and decay times and slopes. The systolic ratio change decreased in cells expressing high biosensor concentration, possibly caused by Ca²⁺ buffering. The GECIs were able to report the effect of nifedipine and propranolol on the heart, which resulted in changes in heart rate, diastolic and systolic Ca²⁺ levels, and Ca²⁺ kinetics. As a result, Twitch-1 and Twitch-4 (Kd 0.25 and 2.8 µM, respectively) seem the most promising GECIs for generating transgenic zebrafish lines, which could be used for modeling heart disorders, for drug screening, and for cardiotoxicity assessment during drug development.

Zebrafish embryos and larvae have emerged as an excellent model in cardiovascular research and are amenable to live imaging with genetically encoded biosensors to study cardiac cell behaviours, including calcium dynamics. To monitor calcium ion levels in three to five days post-fertilization larvae, we have used bioluminescence. We generated a transgenic line expressing GFP-aequorin in the heart, Tg(myl7:GA), and optimized a reconstitution protocol to boost aequorin luminescence. The analogue diacetylh-coelenterazine enhanced light output and signal-to-noise ratio. With this cardioluminescence model, we imaged the time-averaged calcium levels and beat-to-beat calcium oscillations continuously for hours. As a proof-of-concept of the transgenic line, changes in ventricular calcium levels were observed by Bay K8644, an L-type calcium channel activator and with the blocker nifedipine. The β-adrenergic blocker propranolol decreased calcium levels, heart rate, stroke volume, and cardiac output, suggesting that larvae have a basal adrenergic tone. Zebrafish larvae treated with terfenadine for 24 h have been proposed as a model of heart failure. Tg(myl7:GA) larvae treated with terfenadine showed bradycardia, 2:1 atrioventricular block, decreased time-averaged ventricular calcium levels but increased calcium transient amplitude, and reduced cardiac output. As alterations of calcium signalling are involved in the pathogenesis of heart failure and arrhythmia, the GFP-aequorin transgenic line provides a powerful platform for understanding calcium dynamics.

Aim: Bradyarrhythmias result from inhibition of automaticity, prolonged repolarization, or slow conduction in the heart. The ERG channels mediate the repolarizing current I_Kr in the cardiac action potential, whereas T‐type calcium channels (TTCC) are involved in the sinoatrial pacemaker and atrioventricular conduction in mammals. Zebrafish have become a valuable research model for human cardiac electrophysiology and disease. Here, we investigate the contribution of ERG channels and TTCCs to the pacemaker and atrioventricular conduction in zebrafish larvae and determine the mechanisms causing atrioventricular block.Methods: Zebrafish larvae expressing ratiometric fluorescent Ca²⁺ biosensors in the heart were used to measure Ca²⁺ levels and rhythm in beating hearts in vivo, concurrently with contraction and hemodynamics. The atrioventricular delay (the time between the start of atrial and ventricular Ca²⁺ transients) was used to measure impulse conduction velocity and distinguished between slow conduction and prolonged refractoriness as the cause of the conduction block.Results: ERG blockers caused bradycardia and atrioventricular block by prolonging the refractory period in the atrioventricular canal and in working ventricular myocytes. In contrast, inhibition of TTCCs caused bradycardia and second‐degree block (Mobitz type I) by slowing atrioventricular conduction. TTCC block did not affect ventricular contractility, despite being highly expressed in cardiomyocytes. Concomitant measurement of Ca²⁺ levels and ventricular size showed mechano‐mechanical coupling: increased preload resulted in a stronger heart contraction in vivo.Conclusion: ERG channels and TTCCs influence the heart rate and atrioventricular conduction in zebrafish larvae. The zebrafish lines expressing Ca²⁺ biosensors in the heart allow us to investigate physiological feedback mechanisms and complex arrhythmias.