Minireview: Epidemiological impact of arboviral diseases in Latin American countries, arbovirus-vector interactions and control strategies.

Mosquitoes are the most crucial insects in public health due to their vector capacity and competence to transmit pathogens, including arboviruses, bacterias and parasites. Re-emerging and emerging arboviral diseases, such as yellow fever virus (YFV), dengue virus (DENV), zika virus (ZIKV), and chikungunya virus (CHIKV), constitute one of the most critical health public concerns in Latin America. These diseases present a significant incidence within the human settlements increasing morbidity and mortality events. Likewise, among the different genus of mosquito vectors of arboviruses, those of the most significant medical importance corresponds to Aedes and Culex. In Latin America, the mosquito vector species of YFV, DENV, ZIKV, and CHIKV are mainly Aedes aegypti and Ae. Albopictus. Ae. aegypti is recognized as the primary vector in urban environments, whereas Ae. albopictus, recently introduced in the Americas, is more prone to rural settings. This minireview focuses on what is known about the epidemiological impact of mosquito-borne diseases in Latin American countries, with particular emphasis on YFV, DENV, ZIKV and CHIKV, vector mosquitoes, geographic distribution, and vector-arbovirus interactions. Besides, it was analyzed how climate change and social factors have influenced the spread of arboviruses and the control strategies developed against mosquitoes in this continent.

Evaluating the Biological Cycle and Reproductive and Population Parameters of Calliphora vicina (Diptera: Calliphoridae) Reared on Three Different Diets.

This work was aimed at evaluating Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae) life cycle, life tables, and reproductive and population parameters. This species is a necrophagous, synanthropic, cosmopolitan fly with broad distribution in the Neotropical region. Six successive generations from adult specimens collected in Bogotá, Colombia, were analyzed in laboratory conditions. The fly's life cycle was evaluated, using a natural diet of pig's liver (L) and two artificial diets-milk powder and egg (ME) and powdered liver (PL). The species' longest life cycle duration (in days), from egg to pupa, was obtained with the ME diet (24.6 ± 6.7) whereas the lowest value was recorded with the L diet (18.8 ± 3.6). Female adult forms achieved high longevity values, this being highest in ME (102.8 ± 3.6). The most efficient diets (L and ME) led to the following average reproductive and population values: 226.2 and 465.4 net reproductive rate (Ro), 22.2 and 47.1 cohort generation time (Tc), 0.24 and 0.13 intrinsic population growth rate (rm), and 1.27 and 1.13 finite rate of increase (λ). The set of results (in terms of relatively high average values concerning the biological characteristics analyzed) showed the species' adaptability to the environmental and nutritional conditions established in a laboratory setting, thereby enabling their colonization.

Evaluating Sarconesiopsis magellanica blowfly-derived larval therapy and comparing it to Lucilia sericata-derived therapy in an animal model.

Larval therapy is used as alternative treatment for hard-to-heal chronic and infected wounds. Lucilia sericata is the most used blowfly species. However, it has been shown recently that Sarconesiopsis magellanica larval excretions and secretions have potent antibacterial activity; this blowfly belongs to the Calliphoridae family. The present work has dealt with evaluating larval therapy using S. magellanica on wounds induced in diabetic rabbits and its action was compared to the effect induced by L. sericata. Twelve New Zealand White rabbits (Oryctolagus cuniculus) were used; they were divided into 4 groups, the first two being treated with larval therapy derived from both aforementioned necrophagous blowflies, an antibiotic was used in the third and the fourth was used as control. All the animals were wounded on the back and infected with Pseudomonas aeruginosa and Staphylococcus aureus. Samples of the secretion from each animal's infected wound were taken and sown on blood agar. The colony forming units were then counted. The PUSH scale was used for the macroscopic evaluation of the wounds. Bacterial control was encountered 48 h post-treatment in the treatments involving larval therapy and to a lesser extent with the antibiotic. Likewise, wound debridement was quicker and more efficient with larval therapy compared to the antibiotic group; however, wound closing time was 23 days in all treatments. The group treated with S. magellanica larvae had relatively quicker evolution until the proliferation phase and the start of maturation, even though there were no significant differences between both blowfly species evaluated here regarding treatments by the end of the treatment period. The present study has validated the diabetic rabbit model for inducing chronic wounds regarding larval therapy and has likewise confirmed the effectiveness of S. magellanica-derived larval therapy as an alternative for curing and healing wounds.

The effects of Sarconesiopsis magellanica larvae (Diptera: Calliphoridae) excretions and secretions on fibroblasts.

Sarconesiopsis magellanica is a necrophagous blowfly which is relevant in both forensic and medical sciences. Previous studies regarding this species have led to understanding life-cycle, population and reproduction parameters, as well as identifying and characterising proteolytic enzymes derived from larval excretions and secretions (ES). As other studies have shown that ES proteolytic activity plays a significant role in wound healing and fibroblasts play a relevant role in granulation tissue formation during such healing, the present study was aimed at analysing the biological effect of S. magellanica larval ES on fibroblasts. ES were obtained from third-instar larvae and added to fibroblast cells at three concentrations (10, 5 and 1 µg/mL) to evaluate their behaviour. MTT assays were used for analysing cell proliferation and viability, whilst cell adhesion was measured by optical density with 10% SDS. Fibroblast migration and morphology was recorded by microscopic observation. ES did not affect fibroblast viability and induced an increase in cell proliferation; cell adhesion became reduced, whilst cell migration through extracellular matrix increased. ES also induced a decreased cell surface and morphological alterations. Changes in all the above-mentioned parameters were reduced when ES were incubated at 60 °C, probably due to protease denaturation. These results suggested that the proteases contained in S. magellanica larval ES contributed towards granulation tissue formation, increased cell migration and promoted cell proliferation. All these data support carrying out further experiments aimed at validating S. magellanica usefulness in larval therapy.

Establishment and characterisation of a new cell line derived from Culex quinquefasciatus (Diptera: Culicidae).

Insect cell cultures are an important biotechnological tool for basic and applied studies. The objective of this work was to establish and characterise a new cell line from Culex quinquefasciatus embryonic tissues. Embryonated eggs were taken as a source of tissue to make explants that were seeded in L-15, Grace's, Grace's/L-15, MM/VP12, Schneider's and DMEM culture media with a pH range from 6.7-6.9 and incubated at 28ºC. The morphological, cytogenetic, biochemical and molecular characteristics of the cell cultures were examined by observing the cell shapes, obtaining the karyotypes, using a cellulose-acetate electrophoretic system and performing random amplified polymorphic DNA-polymerase chain reaction analysis, respectively. The Grace's/L-15 medium provided the optimal nutritional conditions for cell adhesion and proliferation. Approximately 40-60 days following the explant procedure, a confluent monolayer was formed. Cellular morphology in the primary cultures and the subcultures was heterogeneous, but in the monolayer the epithelioid morphology type predominated. A karyotype with a diploid number of six chromosomes (2n = 6) was observed. Isoenzymatic and molecular patterns of the mosquito cell cultures matched those obtained from the immature and adult forms of the same species. Eighteen subcultures were generated. These cell cultures potentially constitute a useful tool for use in biomedical applications.

Establishment and characterisation of a new cell line derived from Culex quinquefasciatus (Diptera: Culicidae)

Insect cell cultures are an important biotechnological tool for basic and applied studies. The objective of this work was to establish and characterise a new cell line from Culex quinquefasciatus embryonic tissues. Embryonated eggs were taken as a source of tissue to make explants that were seeded in L-15, Grace's, Grace's/L-15, MM/VP12, Schneider's and DMEM culture media with a pH range from 6.7-6.9 and incubated at 28ºC. The morphological, cytogenetic, biochemical and molecular characteristics of the cell cultures were examined by observing the cell shapes, obtaining the karyotypes, using a cellulose-acetate electrophoretic system and performing random amplified polymorphic DNA-polymerase chain reaction analysis, respectively. The Grace's/L-15 medium provided the optimal nutritional conditions for cell adhesion and proliferation. Approximately 40-60 days following the explant procedure, a confluent monolayer was formed. Cellular morphology in the primary cultures and the subcultures was heterogeneous, but in the monolayer the epithelioid morphology type predominated. A karyotype with a diploid number of six chromosomes (2n = 6) was observed. Isoenzymatic and molecular patterns of the mosquito cell cultures matched those obtained from the immature and adult forms of the same species. Eighteen subcultures were generated. These cell cultures potentially constitute a useful tool for use in biomedical applications.

Lucilia sericata strain from Colombia: Experimental colonization, life tables and evaluation of two artificial diets of the blowfly Lucilia sericata (Meigen) (Diptera: Calliphoridae), Bogotá, Colombia strain.

The objective of this work was to establish, under experimental laboratory conditions, a colony of Lucilia sericata, Bogotá-Colombia strain, to build life tables and evaluate two artificial diets. This blowfly is frequently used in postmortem interval studies and in injury treatment. The parental adult insects collected in Bogotá were maintained in cages at 22°C±1 average temperature, 60%±5 relative humidity and 12 h photoperiodicity. The blowflies were fed on two artificial diets that were evaluated over seven continuous generations. Reproductive and population parameters were assessed. The life cycle of the species was expressed in the number of days of the different stages: egg = 0.8±0.1, larvae I = 1.1±0.02, larvae II = 1.94±0.16, larvae III = 3.5±0.54, pupae = 6.55±0.47, male adult = 28.7±0.83 and female adult = 33.5±1.0. Total survival from egg stage to adult stage was 91.2% for diet 1, while for diet 2 this parameter was 40.5%. The lifetime reproductive output was 184.51±11.2 eggs per female. The population parameters, as well as the reproductive output of the blowflies that were assessed, showed relatively high values, giving evidence of the continuous increase of the strain over the different generations and making possible its maintenance as a stable colony that has lasted for more than two years.

Lucilia sericata strain from Colombia: Experimental Colonization, Life Tables and Evaluation of Two Artifcial Diets of the Blowfy Lucilia sericata (Meigen) (Diptera: Calliphoridae), Bogotá, Colombia Strain

The objective of this work was to establish, under experimental laboratory conditions, a colony of Lucilia sericata, Bogotá-Colombia strain, to build life tables and evaluate two artifcial diets. This blowfy is frequently used in postmortem interval studies and in injury treatment. The parental adult insects collected in Bogotá were maintained in cages at 22°C±1 average temperature, 60 percent±5 relative humidity and 12 h photoperiodicity. The blowfies were fed on two artifcial diets that were evaluated over seven continuous generations. Reproductive and population parameters were assessed. The life cycle of the species was expressed in the number of days of the different stages: egg = 0.8±0.1, larvae I = 1.1±0.02, larvae II = 1.94±0.16, larvae III = 3.5±0.54, pupae = 6.55±0.47, male adult = 28.7±0.83 and female adult = 33.5±1.0. Total survival from egg stage to adult stage was 91.2 percent for diet 1, while for diet 2 this parameter was 40.5 percent. The lifetime reproductive output was 184.51±11.2 eggs per female. The population parameters, as well as the reproductive output of the blowfies that were assessed, showed relatively high values, giving evidence of the continuous increase of the strain over the different generations and making possible its maintenance as a stable colony that has lasted for more than two years.