Molecular evidence of pathogens and endosymbionts in the black horse fly Osca lata (Diptera: Tabanidae) in Southern Chile.

Little is known about the role of horse flies in potential pathogen transmission in Chile. This study provides evidence of the molecular detection of microorganisms in southern Chile. In the present study, adult Osca lata horse flies were trapped from Punucapa (39°45'06"S/73°16'08"W, Región de Los Ríos) and Puyehue (40°39'10"S/72°10'57"W, Región de Los Lagos), Chile. Among the 95 samples analyzed by PCR using specific primers, microorganisms were detected in 23.2% (n = 22) of the samples. Rickettsia spp. DNA was detected in 15.8% (n = 15) of the samples, Trypanosomatidae DNA in 5.3% (n = 5) of the samples, and filarial DNA in 2.1% (n = 2) of the samples. This study found that horse flies in the region are capable of carrying a variety of both parasites and endosymbionts. Further research is needed to understand the specific impact of horse flies as mechanical or biological vectors and develop effective control measures to prevent the spread of any microorganisms associated with disease.

Protective Encapsulation of a Bioactive Compound in Starch-Polyethylene Glycol-Modified Microparticles: Degradation Analysis with Enzymes.

Starch is a promising polymer for creating novel microparticulate systems with superior biocompatibility and controlled drug delivery capabilities. In this study, we synthesized polyethylene glycol (PEG)-modified starch microparticles and encapsulated folic acid using a solvent-mediated acid-base precipitation method with magnetic stirring, which is a simple and effective method. To evaluate particle degradation, we simulated physiological conditions by employing an enzymatic degradation approach. Our results with FTIR and SEM confirmed the successful synthesis of starch-PEG microparticles encapsulating folic acid. The average size of starch microparticles encapsulating folic acid was 4.97 µm and increased to 6.01 µm upon modification with PEG. The microparticles were first exposed to amylase at pH 6.7 and pepsin at pH 1.5 at different incubation times at physiological temperature with shaking. Post-degradation analysis revealed changes in particle size and morphology, indicating effective enzymatic degradation. FTIR spectroscopy was used to assess the chemical composition before and after degradation. The initial FTIR spectra displayed characteristic peaks of starch, PEG, and folic acid, which showed decreased intensities after enzymatic degradation, suggesting alterations in chemical composition. These findings demonstrate the ongoing development of starch-PEG microparticles for controlled drug delivery and other biomedical applications and provide the basis for further exploration of PEG-starch as a versatile biomaterial for encapsulating bioactive compounds.

The role of multi-walled carbon nanotubes in enhancing the hydrolysis and thermal stability of PLA.

Polylactic acid (PLA) is a bioresorbable and biodegradable polymer extensively used in various biomedical and engineering applications. In this study, we investigated the mass loss and thermal properties of PLA-multi-walled carbon nanotube (MWCNT) composites under simulated physiological conditions. The composites were prepared by melting PLA with 0.1, 0.5, 1.0, and 5.0 wt% MWCNTs using an ultrasonic agitator, and FTIR analysis confirmed composite formation. Subsequently, the composites were subjected to hydrolysis under simulated physiological conditions (pH 7.4 and 37 °C) for up to 60 days. The results revealed that the mass loss of the composites decreased with increasing MWCNT content, suggesting that the presence of MWCNTs decelerated the hydrolysis process. On day 58, the mass loss of pure PLA was 12.5%, decreasing to 8.34% with 0.1% MWCNT, 5.94% with 0.5% MWCNT, 4.59% with 1% MWCNT, and 3.54% with 5.0% MWCNT. This study offers valuable insights into the behavior of PLA-MWCNT composites under physiologically simulated conditions, facilitating the development of new polymer composites with enhanced thermal stability and degradation resistance for biomedical applications.

Unraveling biotypes of the northern house mosquito, Culex pipiens s.l. (Diptera: Culicidae): molecular differentiation and morphometric analysis.

Geometric morphometrics was used to determine whether geographic isolation could explain differences in wing size and shape between and within continental (27°S to 41°S) and insular (Rapa Nui) populations of Culex pipiens s.s. Linnaeus and their biotypes (f. pipiens and f. molestus). Molecular protocols based on polymorphisms in the second intron of nuclear locus ace-2 (acetylcholinesterase-2) were used to differentiate Cx. pipiens s.s. from Cx. quinquefasciatus Say, and an assay based on polymorphisms in the flanking region of a microsatellite locus (CQ11) was used to identify biotypes. Culex pipiens f. molestus and hybrids shared larval habitats in all continental sites, while Cx. pipiens f. pipiens was found in 5 of the 10 sites. Only biotype molestus was found in Rapa Nui (Easter Island) Pipiens and molestus biotypes occur sympatrically in aboveground locations, and only molestus was found in the underground site (ME). Biotype molestus was dominant in rural locations and preferably anthropophilic. These results agree with the ecological descriptions previously reported for the biotypes of Cx. pipiens s.s. Procrustes ANOVA only showed differences in centroid size between biotypes in females and males and did not show significant differences in wing shape. However, we found significant differences among the geographic areas in the centroid size and wing shape of both females and males. Particularly, the population of Rapa Nui Island had shorter wings than the continental populations. The results highlight the effects of geographic and environmental processes on morphotypes in vector mosquitoes.

Nanoparticle-Mediated Delivery of Flavonoids: Impact on Proinflammatory Cytokine Production: A Systematic Review.

Flavonoids are a diverse group of plant-derived compounds that have been shown to have various health benefits, including anti-inflammatory effects. However, their use in the treatment of inflammatory diseases has been limited due to their low bioavailability. The nanoparticle-mediated delivery of flavonoids has been proposed as a potential solution to this issue, as it allows the sustained release of the flavonoids over time. There are several different nanoparticle systems that have been developed for flavonoid delivery, including polymeric nanoparticles, liposomes, and inorganic nanoparticles. This systematic review aims to evaluate the impact of nanoparticle-mediated delivery of flavonoids on pro-inflammatory cytokine production in various diseases. We analyzed the performance of flavonoid-encapsulated nanoparticles in regulating cytokine production in different in vitro and in vivo studies. To this end, we followed the guidelines of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) to conduct a comprehensive search of the literature and to assess the quality of the included studies. The results showed that flavonoid-encapsulated nanoparticles significantly downregulated pro-inflammatory cytokines, such as TNF-α, IL-1ß, IL-6, and IL-18. In some cases, this effect was significantly greater than that observed with non-encapsulated flavonoids These findings suggest that nanoparticle-mediated delivery of flavonoids may have potential as a therapeutic approach for the treatment of inflammatory diseases.

On the taxonomic status of Agelanius antenninus (Philip) (Diptera: Tabanidae) from Peru based upon evidence from a morphological analysis of the holotype.

The taxonomic status of Agelanius antenninus (Philip, 1969) was evaluated on the basis of a morphological study of the holotype, described from Huánuco, Peru. Results of analyses of A. antenninus and its morphological comparison with A. meridianus Rondani, the type-species of Agelanius Rondani, proved that A. antenninus does not belong to the genus Agelanius. The subgenus Styphocera Philip is reinstated as a valid subgenus of Stypommisa Enderlein. Thus, the generic combination Stypommisa (Styphocera) antennina Philip should be reinstated for this species.

Unusual presentation of orbital compartment syndrome due to complicated herpes zoster ophthalmicus: A case report.

Herpesvirus reactivates from a latent infection in older adults and critically ill and immunocompromised individuals. Herpes zoster ophthalmicus (HZO) is a latent infection that affects the fifth cranial nerve. It is an infrequent cause of increased intraocular pressure. We present the case of a 50-year-old man with a reactivation of latent varicella-zoster virus infection involving the ophthalmic branch of the fifth cranial nerve. The patient was initially managed as an outpatient with an antiviral, but his clinical evolution worsened and required urgent surgical decompression. Lateral canthotomy was performed with cantholysis of the inferior crus of the lateral canthal tendon. Only partial decompression was achieved, so cantholysis of the upper crus was performed with significant tissue tension release. The patient evolved well and was discharged after 6 days without symptoms for outpatient management.

Effect of Physiological Fluid on the Photothermal Properties of Gold Nanostructured.

Colloidal gold particles have been extensively studied for their potential in hyperthermia treatment due to their ability to become excited in the presence of an external laser. However, their light-to-heat efficiency is affected by the physiologic environment. In this study, we aimed to evaluate the ability of gold sphere, rod, and star-shaped colloids to elevate the temperature of blood plasma and breast cancer-simulated fluid under laser stimulation. Additionally, the dependence of optical properties and colloid stability of gold nanostructures with physiological medium, particle shape, and coating was determined. The light-to-heat efficiency of the gold particle is shape-dependent. The light-to-heat conversion efficiency of a star-shaped colloid is 36% higher than that of sphere-shaped colloids. However, the raised temperature of the surrounding medium is the lowest in the star-shaped colloid. When gold nanostructures are exited with a laser stimulation in a physiological fluid, the ions/cations attach to the surface of the gold particles, resulting in colloidal instability, which limits electron oscillation and diminishes the energy generated by the plasmonic excitation. Fluorescein (Fl) and polyethylene glycol (PEG) attached to gold spheres enhances their colloidal stability and light-to-heat efficiency; post-treatment, they remand their optical properties.

Northern and central Chile still free of emerging flaviviruses in mosquitoes (Diptera: Culicidae).

Geographic isolation and strict control limits in border areas have kept Chile free from various pathogens, including Flavivirus. However, the scenario is changing mainly due to climate change, the reintroduction of more aggressive mosquitoes, and the great wave of migration of people from endemic countries in recent years. Hence, it is necessary to surveillance mosquitoes to anticipate a possible outbreak in the population and take action to control it. This study aimed to investigate the presence of Flavivirus RNA by molecular tools with consensus primers in mosquitoes collected in the extreme north and central Chile. From 2019 to 2021, a prospective study was carried out in localities of Northern and part of Central Chile. Larvae, pupae, and adults of mosquitoes were collected in rural and urban sites in each locality. The collected samples were pooled by species and geographical location and tested using RT-PCR and RT-qPCR to determine presence of Flavivirus. 3085 specimens were collected, the most abundant specie Culex quinquefasciatus in the North and Aedes (Ochlerotatus) albifasciatus in the Center of Chile. Both genera are associated with Flavivirus transmission. However, PCR and RT-PCR did not detect Flavivirus RNA in the mosquitoes studied. These negative results indicate we are still a free Flavivirus country, which is reaffirmed by the non-existence of endemic human cases. Despite this, routine surveillance of mosquitoes and the pathogens they carry is highly recommended to evaluate each area-specific risk of vector-borne transmission.

Revalidation of Phalangomyia Dyar & Knab as a subgenus of Culex L. (Diptera: Culicidae) based on morphological and molecular evidence.

Culex apicinus Philippi, 1865 and its synonyms, Phalangomyia debilis Dyar & Knab, 1914, Cx. escomeli Brèthes, 1920 and Cx. alticola Martini, 1931, were re-examined employing molecular and morphological phylogenetic methods. Archival and fresh specimens were used for morphological diagnosis and to generate fragments of two single-copy nuclear genes (CAD, HB) and one mitochondrial gene (COI). Additional sequences of Cx. apicinus and species of the subgenus Culex of Culex Linnaeus, 1758 and other subgenera of the genus were obtained from GenBank for molecular analysis. Diagnostic morphological characteristics of Cx. apicinus and other species of the subgenus Culex were contrasted. Phylogenetic relationships were generated using DNA sequences of protein coding genes under both Maximum Likelihood and Bayesian approaches. Culex apicinus showed unique morphological traits that allow its classification in a separate subgenus. The subspecific status is supported by its recovery as a monophyletic clade within the genus Culex based on morphological and molecular characters. The synonymous Phalangomyia debilis provides an available subgeneric name. Consequently, Phalangomyia Dyar & Knab, 1914 is resurrected from synonymy with Culex Linnaeus, 1758 as a monobasic subgenus of Culex.

Gold Nanoparticles as Drug Carriers: The Role of Silica and PEG as Surface Coatings in Optimizing Drug Loading.

The use of gold nanoparticles as drug delivery systems has received increasing attention due to their unique properties, such as their high stability and biocompatibility. However, gold nanoparticles have a high affinity for proteins, which can result in their rapid clearance from the body and limited drug loading capabilities. To address these limitations, we coated the gold nanoparticles with silica and PEG, which are known to improve the stability of nanoparticles. The synthesis of the nanoparticles was carried out using a reduction method. The nanoparticles' size, morphology, and drug loading capacity were also studied. The SEM images showed a spherical and homogeneous morphology; they also showed that the coatings increased the average size of the nanoparticles. The results of this study provide insight into the potential of gold nanoparticles coated with silica and PEG as drug delivery systems. We used ibuprofen as a model drug and found that the highest drug load occurred in PEG-coated nanoparticles and then in silica-coated nanoparticles, while the uncoated nanoparticles had a lower drug loading capacity. The coatings were found to significantly improve the stability and drug load properties of the nanoparticles, making them promising candidates for further development as targeted and controlled release drug delivery systems.

Lipid nanoparticles for gene therapy in ocular diseases.

OBJECTIVES: Lipid nanoparticles, as a nucleic acid delivery system, have been used as an alternative to treat ocular diseases, since they can cross the ocular barrier and efficiently transfecting nucleic acids to various cells of the eye. The size influences the transfection of genes, biological distribution, diffusion, and cellular uptake. It is therefore important to establish a relationship between size, formulation, and encapsulation percentage. EVIDENCE ACQUISITION: In this review, we used a search strategy to compare studies of nanomedicine systems aimed at eye diseases where the size of the nanoparticles and the efficiency of encapsulation of genetic material are reported based on the criteria of Preferred Reporting Items for Systematic Reviews (PRISMA ScR 2020 guidelines). RESULTS: Out of the initial 5932, 169 studies met the inclusion criteria and were included to form the basis of the analysis. Nanoparticles reported are composed mainly of PEG-modified lipids, cholesterol, and cationic lipids, that in combination with messenger or interference RNA, allow the formulation of a nanoparticle with an encapsulation efficiency greater than 95%. The diseases treated mainly focus on conditions related to the retina and cornea. Certain characteristics of nanoparticles increase encapsulation efficiency, such as the size of the nanoparticle and the charge of the outer layer of the nanoparticle. CONCLUSION: It is still unknown what characteristics lipid nanoparticles should have to successfully treat human eye illnesses. The in vitro and in vivo investigations covered in this review, however, present encouraging results. To improve encapsulation effectiveness and disease gene silencing, nanoparticle formulation is essential. The most stable nanoparticles are those made mostly of cationic lipids, PEG lipids, and cholesterol, which also effectively encapsulate RNA. The encapsulation efficiency is not only influenced by size, but also by other factors such as methods of preparation.

Citizen Science Meet South American Tachinids: New Records of Feather-Legged Fly Trichopoda (Galactomyia) pictipennis Bigot (Diptera: Tachinidae) from Chile.

The species of Gymnosomatini (Diptera: Tachinidae) are specialized to attack stink bugs (Hemiptera: Pentatomidae). Some of these flies, such as those in the genus Trichopoda Berthold, are conspicuous and represent target species to study with citizen science. Here we report for the first time the distribution range of T. (Galactomyia) pictipennis Bigot, a newly introduced biological control species of tachinid for use against stink bugs in Chile using data obtained through the citizen science method. Additionally, we update the distribution of T. arcuata (Bigot), a native biological control species of tachinid, and assess the grade of overlap between the flies and their main stink bug hosts. We obtained data regarding the occurrence of flies and stink bugs from the citizen science program "Moscas Florícolas de Chile," from Facebook groups, and from iNaturalist. We standardized our data to avoid temporal, spatial, and geographic biases. We report the extent of occurrence (EOO) and area of occupancy (AOO) for each fly and stink bug species. The EOOs for T. pictipennis and T. arcuata are 85,474 km2 and 20,647 km2 through central Chile ecosystems, respectively. The EOO of T. arcuata is overlapped by 53% with the introduced species. Trichopoda pictipennis has a high percentage of overlap with the EOO of Nezara viridula Linnaeus (61%), whereas T. arcuata has low percentages of overlap with different native Acledra species and with N. viridula. We discuss the potential of citizen science to detect (1) areas with higher diversity or gaps of occurrences, (2) new biological control agents, (3) prey records, (4) negative impacts on non-target species, (5) changes in composition at long-term, and (6) areas to promote conservation biological control in agricultural landscapes.

A catalog of Pipunculidae of Chile (Diptera)

An updated catalog of the Pipunculidae from Chile is provided. We present all valid names and synonyms for the 21 species belonging to six genera and three subfamilies distributed in the country. Information is given on name, author, year of publication, page number, type species, type depository, type locality, Chilean localities, and references. Tomosvaryella Aczél and Eudorylas Aczél are the richest genera in Chile, with eight species each, followed by Protonephrocerus Collin with two species, and Chalarus Walker, Neocephalosphaera De Meyer and Elmohardyia Rafael with one species each. The geographical distribution of the species was compiled from the literature and the retrieval of distributional data from collections.

Filarial nematodes in domestic dogs and mosquitoes (Diptera: Culicidae) from semi-rural areas in Central Chile.

Climate change, competent vectors, and reservoir animals are the main factors for developing vector-borne zoonotic diseases. These diseases encompass a significant and widespread category of pathogens (e.g., viruses, bacteria, protozoa, and helminths) transmitted by blood-feeding arthropods, including ticks, fleas, lice, triatomines, mosquitoes, sandflies, and blackflies. In Chile, several studies have explored the role of dogs as reservoirs of vector-borne pathogens; however, there is a lack of research investigating the presence of pathogens in arthropods. Specifically, within the order Diptera, limited knowledge exists regarding their roles as carriers of pathogens. This study aimed to examine the presence of zoonotic filarial nematodes in mosquitoes and dogs within a previously unstudied semi-rural area of Central Chile. Two hundred samples of dog blood and seven hundred and twenty-four mosquitoes were collected during 2021-2022 and studied for filarial nematodes by PCR. The prevalence of microfilaremic dogs detected by Knott's test was 7.5%, with Acanthocheilonema reconditum being the only species identified. Aedes (Ochlerotatus) albifasciatus was the most abundant mosquito species collected, and 15 out of 65 pools were positive for filarial nematodes. Among these pools, 13 tested positive for Acanthocheilonema reconditum, and two tested positive for Setaria equina through PCR. Additionally, five Culex pipiens specimens were positive for Acanthocheilonema reconditum. Despite the absence of zoonotic filarial species, these findings underscore the significance of monitoring pathogens in mosquitoes and animal hosts and continued research into the dynamics of vector-borne diseases, particularly in unexplored regions.

Sialolitiasis, sialoadenitis y sialoadenosis. Revisión de literatura y reporte de un caso clínico / Sialolithiasis, sialadenitis and sialadenosis. Review of literature and report of a clinical case

Introducción: la formación de sialolitos se considera la alteración más común en las glándulas salivales. Su origen aún no es claro en cuanto a qué provoca la obstrucción parcial o total del sistema de conductos salivales, con una predilección significativa en las glán - dulas submandibulares, lo que permite un desplazamiento retrógrado de bacterias a través del conducto salival hacia la glándula afectada, el cual se traducirá clínicamente como inflamación unilateral o bila - teral. El tratamiento para la sialolitiasis y sialoadenitis puede variar desde un manejo conservador hasta tratamientos que implican la remoción de la glándula salival afectada. La sialoadenosis difiere de las anteriores en su origen, ya que ésta no se considera inflamatoria ni neoplásica y no afecta la función glandular de excretar saliva. Presentación de caso clínico: se trata de paciente masculino con diagnósticos presuntivos de sialoadenitis y sialolitiasis, al cual se le realizó biopsia escisional de la glándula afectada, su estudio histo- patológico corroboró los diagnósticos mencionados anteriormente que además confirmó un diagnóstico de sialoadenosis. Conclusio- nes: la escisión de la glándula salival involucrada se realizará en casos diagnosticados de sialoadenitis y sialolitiasis con sialolitos de gran tamaño y procesos supurativos crónicos que no resuelven con antibioticoterapia (AU)

Convalescent Plasma to Treat COVID-19: A Two-Center, Randomized, Double-Blind Clinical Trial.

Background: The use of convalescent plasma (CP) has been considered for its immunological mechanisms that could benefit patients in moderate and severe stages of COVID-19. This study evaluated the safety and efficacy of the use of donor CP for COVID-19. Material and methods: A double-blind, randomized controlled clinical trial was conducted from May to October 2020. Thirty-nine participants with moderate (II) and severe (III) stages of COVID-19 confirmed by RT-PCR were included. The study randomization rate was set at 3:1. CPs were chosen for application with a neutralizing antibody titer of ≥1:32. Results: We observed a significantly lower 21-day post-transfusion mortality HR: 0.17 (95.0% CI [0.07−0.45, p < 0.001]) in the group receiving CP compared with the control group; protective units (PU) in the group receiving convalescent plasma after seven days were significantly higher (512 (32−16,384) vs. 96 (32−256), p = 0.01); the PAO2/FIO2 index showed a significant improvement in the group receiving CP (251.01 (109.4) vs. 109.2 (62.4), p < 0.001, in the control group). Conclusion: CP is safe and effective, as it decreased mortality in the CP group compared with the control group.

Experimental data validating the optimization of a wireless power transfer prototype employing a novel phase shift measurement system and frequency control.

Resonant wireless power transfer (WPT) systems have been evolving and improving their designs over the last few years, looking to efficiently charge electric vehicles, cellphones, and biomedical devices. In this article, we present to the scientific community the data obtained from the optimization of a resonant WPT prototype, operating at different vertical misalignments and load conditions, known to have an impact on the behavior of these type of systems. To maximize the power transferred to the load, we developed a proportional-integral frequency control algorithm that employs the phase-shift between the voltage and current waveforms in the transmitting antenna (resonance indicator) as a setpoint. Data on the performance and control optimization process of the prototype during laboratory tests were acquired using a LabVIEW interface, which was designed to capture information such as the evolution of the frequency, the phase-shift, and the load voltage, from multiple devices (a microcontroller, an oscilloscope, a digital multimeter, and a controllable power supply). The data were organized and presented in tables and graphs using MATLAB. The importance of the dataset relies on the opportunity to utilize the information as a basis for the improvement of the associated electronics by using different transmission topologies, higher speed components, new-generation microcontrollers, and to modelling novel intelligent control algorithms, such as adaptative neuro-fuzzy inference systems.

Chagas disease in northern Chile: Detection of Trypanosoma cruzi in children, dogs and triatomine bugs.

Chagas disease is an anthropozoonotic disease caused by the protozoan Trypanosoma cruzi, transmitted by triatomine vectors. In Chile, there are four species of triatomine bugs that are potential vectors of T. cruzi, being Triatoma infestans the main vector in endemic areas of the country. The "Programa Nacional de Control Vectorial de la Enfermedad de Chagas de Chile" has significantly reduced the rates of home infestation to less than 1% and has interrupted vectorial transmission since 1999. The objective of this study was to evaluate the impact of vectorial control and the continuity of the interruption of vectorial transmission in northern Chile (provincia de El Loa, región de Antofagasta). The study comprised fingerstick blood samples of 2104 children, attending local school, venous blood samples of 65 dogs, associated to houses with T. infestans unique findings and vector infestation, and intestine samples of 284 T. infestans specimens, from the provincia de El Loa, during 2014-2016 period. The samples were analyzed by enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence and/or polymerase chain reaction (PCR) techniques. A total of 5 children (0.24%), 7 dogs (10.8%), and 6 specimens of T. infestans (2.1%) resulted positive to T. cruzi infection. This study showed that the risk of transmission of Chagas disease is low in the north of Chile (provincia de El Loa), detected a low positive rate of chagasic children and of infected triatomine bugs, and showed the existence of T. cruzi transmission in dogs, which are used as natural sentinels for the detection of T. cruzi infection, being especially useful during surveillance program in human population characterized by low seroprevalence.

Improving geographic distribution data for a putatively extinct species, a test case with a disappeared fly.

Global biodiversity loss by anthropogenic impacts is an under-recognized form of global environmental change. Global defaunation is still poorly documented in the case of insects, showing a significant decrease in populations and diversity. The blowfly Neta chilensis (Walker 1837) is poorly known and presumed to be confined to southern-South America, with an unclear distributional pattern. It was last collected in 1984. We aimed, through Ecological Niche Models, to identify regions highly suitable for N. chilensis; to test the suitability of regions with doubtful records; to understand the impact that climatic change and human activities have had; and to identify regions with high chances to find it. We compiled 130 presence records from Argentina and Chile and 117 localities where it was sought but not found between 1987 and 2018. Results indicate that suitable conditions are restricted to southern and central Chile and to southwestern Argentina, that doubtful records are predicted in unsuitable areas, that N. chilensis occupies a narrow niche and that its decline is not mainly caused by climate changes but more probably to habitat loss and to the biological invasion. We identified two regions where the chances of finding the species are higher in the case that it is not extinct already.