Lactuca L. species belong to the Asteraceae family and these plants are traditionally used for therapeutic purposes around the world. The dried milky latex of L. serriola is known as “lettuce oil” and is used as a sedative in Turkey. This study aimed to evaluate the sedative effects and analyze the chemical compositions of latexes obtained from some Lactuca species growing in Turkey. The sedative effects were evaluated through various behavioral tests on mice. For this purpose, latexes were obtained from L. glareosa Boiss., L. viminea (L.) J. Presl and C. P, L. mulgedioides (Vis and Panćić) Boiss. and Kotschy ex. Boiss., L. saligna L., and L. serriola L. The latex from L. saligna showed the highest sedative effects, whilst L. serriola and L. viminea latexes also displayed significant sedative effects compared to the control group at a dose of 100 mg/kg. However, the latexes from L. glareosa and L. mulqedioides did not exhibit any sedative effects on mice. Characteristic sesquiterpene lactones  were determined qualitatively and quantitatively by high-performance liquid chromatography (HPLC). Lactucin was identified as the main component. Sleep is the state of rest that is necessary for all people to lead healthy lives. Many physical, environmental, psychological, and physiological factors can positively or negatively affect the quality and quantity of sleep. Insomnia is considered an important health problem because of its negative effects on people’s quality of life and it often has psychiatric or medical causes . ChronicLactuca is a taxon that contains important species and this genus is widely distributed throughout the world and has been consumed by people since ancient times. L. biennis (Moench) Fernald is used as an analgesic, antidiarrheic, antiemetic, and antihemorrhagic, as well as for heart diseases and gynecological diseases; L. canadensis L. is used as an analgesic, sedative, and stimulant and treats eye and kidney disorders, orthopedic diseases, skin diseases, and warts. L. intybacea Jacq. ex Murray is applied as a decongestant, laxative, tonic, and depurative, and treats arthritis, hepatitis, and gout. L. muralis (L.) Fresen is used as a narcotic, sedative, and antispasmodic. L. sativa L. is taken as an aphrodisiac, sedative, cardioactive, diuretic, hypnotic, narcotic, and sedative, and it treats asthma, fever, and hyperglycemia. L. indica L. is used for its antibacterial and anti-inflammatory effects. L. capensis Thunb is used for treating sores, ulcers, leprosy, eczema, and the milky latex of L. virosa L., known as “lactucarium”, is taken as a sedative-hypnotic [Lactuca species have shown that these plants have analgesic, anti-inflammatory, antioxidant, anticarcinogenic, hepatoprotective, neuroprotective, antidiabetic, and anxiolytic effects [Lactuca species. In particular, lactucine-type guaianolides are the most representative constituents [hypnotic ,6,7. Bio effects ,11,12,13tituents .L. virosa latex and they were specifically responsible for the sedative-hypnotic effects of lactucarium. Several studies on lactucin and its derivatives have confirmed that these compounds have sedative effects. In addition, it has been noted that lactucin and lactucopicrin are active substances that promote sleep, because they bind effectively to the GABAA receptor [Lactucin and its esters were first found in receptor ,16.Lactuca species grown in Turkey. The sedative effects of the latexes obtained from L. serriola, L saligna, L viminea, L. mulgedioides, and L. glareosa were investigated using traction, fireplace, holeboard, and thiopental-induced sleeping tests. In addition, the presence and amounts of the sesquiterpene lactones lactucin and lactucopicrin and their dihydro derivatives were determined by high-performance liquid chromatography (HPLC). This is the first report on the sedative effects of latexes obtained from these Lactuca species.The aim of this study was to evaluate the sedative effects of milky latexes obtained from five L. virosa, L. serriola) has traditionally been used as a sedative. In Turkey, the dried milky latex of L. serriola is known as “lettuce oil” and is used as a sedative [Lactuca species grown in Turkey were investigated by various in vivo methods on mice. The latexes were also analyzed using high-performance liquid chromatography for characteristic compounds thought to be responsible for this effect.In Europe, the dried milky latex of wild lettuce , which are characteristic constituents of the eriments ,15,16. TL. serriola (57.53249 ± 0.27 mgstd/glatex), followed by L. saligna, L. viminea, and L. glareosa in descending order. Lactucopicrin was only detected in the latex from L. saligna (0.61729 ± 0.02 mg std/glatex) and all the other reference compounds were also found in the latex from L. saligna  whilst the highest level of 11β,13-dihydrolactucopicrin was seen in the latex from L. glareosa (0.75238 ± 0.01 mg std/glatex). In L. mulgedioides, none of the compounds of interest was detected.Sesquiterpene lactones were identified by retention times. Quantification was achieved by the measurement of the peak area at 260 nm and by comparison with standard calibration curves . The hig saligna . The 11βL. serriola was considerably higher than the amount of lactucin detected in L. saligna, the sedative effect of the latex from L. serriola was lower than that of the latex from L. saligna.Although the amount of lactucin detected in Lactuca species are used in folk medicine for stomach problems, pain relief, and inflammation. Furthermore, it has been reported to have anticonvulsant, sedative-hypnotic, and antioxidant properties [L. sativa displayed a remarkable sedative effect [L. sativa enhanced sleeping time and acted via the GABAergic system [operties . Sutrisne effect . Additioc system .Lactuca genus. However, few studies have been reported with latexes from Lactuca species. The latexes of L. mulgedioides, L. saligna, L. viminea, and L. glareosa were examined for the first time in this study. Sessa et al. noted that important information about the sesquiterpene lactone content of the latex of L. sativa, was obtained and they reported that the major sesquiterpene lactones of Lactuca species are 15-oxalyl and 8-sulfate conjugates of guaianolides, such as lactucin, deoxylactucin, and lactucopicrin [In the literature, the quantitative analysis of sesquiterpene lactones has been reported for various species of copicrin . In thisL. sativa and our plant material, L. serriola [The literature shows that different amounts of latex are found in different vegetation stages of the plant. It has been reported that the sesquiterpene lactone content is highest during bolting. However, in our study, all the latexes were collected during the flowering stage because the amount of latexes of plants was high at this stage. The lactucin amounts were compared with the literature and a significant difference was observed between serriola . The resL. sativa were investigated using the pentobarbital-induced sleeping test by Kim et al. and the amounts of lactucin and lactucopicrin in the extracts were determined by HPLC. A stronger sedative effect was observed in the seed extract of L. sativa which contained more lactucopicrin [The sedative effects of 70% ethanolic extracts of the leaves and seeds of different varieties of copicrin . In anotcopicrin . In addicopicrin .L. serriola (LSE) whose lactucin content was significantly higher  had no higher sedative effect. The lactucopicrin compound was found only in L. saligna (LSA) and a high sedative effect was observed in this latex. In L. viminea (LM), no components we analyzed were seen in this latex and accordingly no sedative effect was obtained. These results confirm that lactucin-type guaianolides are responsible for the sedative effects.According to our study results, in order to understand the mechanism of the effects of the compounds, further studies should be carried out. Our results showed that the sedative effect is not caused by lactucin, because the latex of We can understand from these results that lactucin and lactucopicrin likely have a synergistic effect in increasing sedative activity. Moreover, other components of latexes that we did not detect may also be responsible for the sedative effect. Therefore, in-depth structural determination and characterization studies should be carried out for all of the main components of latex, which will help elucidate the compound or compounds responsible for the sedative effect.Lactuca species were collected from different habitats in Central Anatolia during the flowering period from July to September of 2018 for the extraction of latex. The obtained plants  were authenticated by Prof. Dr. Ergin Hamzaoglu from Gazi University.Firstly, various The stems of the plants were wounded with a scalpel and the milky latex was released onto the plant surface. The latex was then collected in glass bottles. This process was repeated three times and the samples were stored at –20 °C until the moment of the analysis.g for 10 min. The obtained supernatant was then filtered through a 0.45 µm membrane filter. The extracted latexes were analyzed by liquid chromatography using an Agilent 110 .The collected latex samples were extracted with methanol and centrifuged at 16,000× 18 column  with a CH3CN/H2O gradient over 40 min at 1 mL min−1 and 40 °C. The solvent mixture consisted of acetonitrile (solvent A) and water (solvent B). The portion of A was increased from 12% to 95% in 37 min gradually, then returned to the initial conditions in 2 min and the percentage of B was decreased from 88% to 5% in 37 min and then returned to the initial conditions.Samples were injected into a reverse-phase (RP) CMale BALB/c mice (25–30 g) were obtained from Kobay Animals Laboratory . The animals were allowed to adapt to the laboratory conditions for 3 days before the experiment. In this process, the animals were housed with a 12-h light, 12-h dark cycle with standard laboratory chow and tap water.Each test group consisted of six mice and all the studies were performed conferring to the international rules regarding the animal experiments and biodiversity rights .In all the tests, samples were administered intraperitoneally in a 0.5% sodium carboxymethylcellulose (CMC) suspension at a dose of 100 mg/kg. In the control group, only 0.5% CMC was administered.Lorazepam  was used as a reference drug and it was administered to the mice intraperitoneally at a dose of 1 mg/kg.The traction test was applied to the mice to evaluate the sedative effects of the test materials using the method described by Courvoisier with Laroche and Rousselet ,22,23,24The test samples and lorazepam as a reference material were administered to the mice intraperitoneally. An hour later, the mice were placed horizontally with their forepaws on a taut rope. Normally a mouse hanging on the rope will lift its hind legs, whilst mice that fail to lift at least one of their hind legs to reach the rope are considered to be sedated. In addition, the behavior of the animals was recorded during the experimental period.The method developed by Hoffman was applied to the mice ,26.The test samples and lorazepam as a reference material were administered to the mice intraperitoneally. An hour later, each mouse was placed in a 30-cm long vertical glass tube. Typically, a normal mouse attempts to escape within thirty seconds, whilst mice that do not make any attempt during this period are considered to be sedated.The method developed by Clark et al. and File and Hoffman was applied to the mice ,28.The test samples and lorazepam as a reference material were administered intraperitoneally to the mice. An hour later, the mice were placed in the center of a perforated board. The perforated board test was carried out using a wooden board of 40 × 40 × 25 cm with evenly spaced holes. The number of times the mice inserted their heads into the holes was recorded.The method developed by Aziz and Khan was applied to the mice ,30,31.Thiopental (a sub-hypnotic dose) was administered to the mice (60 mg/kg) intraperitoneally, then after 30 min the mice were treated with the test samples. The time from the application of the test samples to sleep and the time from sleep to waking was recorded.p < 0.05, ** p < 0.01, and *** p < 0.001.The data obtained from the animal experiments were evaluated using one-way ANOVA statistical tests. In addition, the test and control groups were assessed using Student–Newman–Keuls post hoc tests. The results were compared with those from the control and reference groups. Statistical significance is expressed as *


Ureteral stenosis after percutaneous microwave ablation (MWA) of renal tumor is a rare but severe complication, and its risk factors are not apparent.This study aimed to investigate the risk factors for stenosis of ureter after MWA treatment of the renal tumor that is a rare complication.P < 0.05 is considered significant.Data of 211 patients who underwent MWA for the treatment of renal tumor were retrospectively analyzed from September 2006 to August 2019. Demographic characteristics, clinical features, ablation parameters, and outcomes were analyzed to find out the potential risk factors of this complication. P = 0.225) or ablation zone (P = 0.089) is not related to this complication. Postoperative urine routine  and R.E.N.A.L. score (P < 0.001) is related to this complication. But after multivariate logistic analysis, only R.E.N.A.L. score (P = 0.004) is associated with this complication. The location and growth pattern of tumor and the energy of ablation were not related to this complication independently.Six of 211 patients developed ureter stenosis, and the rate of this complication is 2.84%. The median time of emergence of hydronephrosis was 226  days. Univariate analysis shows the distance between ureter and tumor (The stenosis of the ureter after MWA of renal tumor is not associated with the tumor size, location, or the distance between the ureter and tumor and ablation site independently. But R.E.N.A.L. score is associated with ureter stenosis after MWA for the treatment of renal tumor, which combines the information of location, depth, and size of tumor. Preoperative evaluation of the tumor is necessary for avoiding ureter stenosis. Further studies should focus on these risk factors of this complication. The renal tumor is one of the fatal urological malignancies. The incidence of the renal tumor has been growing. However, the survival rate of the renal tumor has increased , 2. AlthMicrowave ablation (MWA) for the renal tumor is a minimally invasive therapy, which can be performed under the guidance of ultrasound (US) or computed tomography (CT) during the operation or percutaneously directly. It has been proved in the previous study that MWA can achieve a similar effect and lower rate of complication compared with laparoscopic radical nephrectomy and open radical nephrectomy –8. The rThis study is approved by institutional review board at Chinese PLA General Hospital. This retrospectively study enrolled the 211 patients in the Interventional Ultrasound Department of Chinese PLA General Hospital from September 2006 to August 2019 who had undergone MWA for the treatment of renal tumor. All the patients had signed the informed consent form. The institutional database was queried to identify incident patients and collect baseline clinical data including age, sex, comorbidity, lesion location, ablation time, ablation power, the maximum diameter of the tumor and ablation zone, preoperative, postoperative and follow-up imaging examination, and laboratory examination. Our study was approved by the institutional review board. The collected criteria were as follows: (1) conformed to the treatment guidelines of NCCN , (2) refAll the patients are treated with percutaneous MWA under US guidance by experienced doctors. The microwave unit  is capable of producing 100 W of power at 2,450 MHz. An automatic biopsy gun with an 18-gage cutting needle to puncture the biopsy for two to three times to achieve tumor tissue was used, followed by 1% lidocaine local anesthesia . A protective temperature-measured device was inserted to control the temperature of the surrounding tissue. The antenna was then inserted into the mass and placed at a proper location under US guidance. After antennas were placed, intravenous anesthesia was administered by a combination of propofol , and ketamine  via the peripheral vein.Hydrodissection technique is a protective measure to reduce the heat injury of surrounding tissues, and this method has proven its efficiency to avoid the damage to the intestinal tract and renal sinus around –16. For Preoperative imaging examination, such as US, contrast-enhanced US, and contrast-enhanced CT/magnetic resonance imaging (MRI), was retrospectively analyzed to determine the tumor location, the diameter of the tumor, and the relationship between the lesion and adjacent structure. The distance between the ureter and tumor or ablation site was measured on MRI/CT. Postoperative first urine routine was collected to analyze potential risk factors. Postoperative imaging examination was retrieved to measure the diameter of the ablation zone and judge if there was stenosis of ureter and secondary hydronephrosis and record the occurrence time of complication. For patient 2, hydronephrosis was detected by US 180 days after ablation, and she accepted percutaneous nephrostomy . Patient2 test or Fisher exact test for categorical variables and Mann–Whitney U test for continuous variables. Variables in which P value is less than 0.2 or clinically considered meaningful were included into multivariable logistic regression. The statistical analysis was calculated by SPSS 18.0 software package  and R (version 3.6.1). P < 0.05 was considered significant. The receiver operating characteristic curve was plotted by R (version 3.6.1).The data were expressed as median or mean ± SD. The correlation between this complication and clinical characteristics was analyzed using a χSix of 211 patients developed ureter stenosis, and the rate of this complication is 2.84%. The characteristic baseline is given in U test P = 0.093, 0.099), ablation power and time , total energy (P = 0.739), and postoperative urine white blood cell  were unrelated to this complication. The distance between ureter and tumor or ablation zone  was unrelated to this complication. Increased postoperative urine routine  and R.E.N.A.L. score (Mann–Whitney U test P < 0.001) were related to this complication.P < 0.001), diameter of tumor (P = 0.093), and ablation zone (P = 0.123), postoperative urine routine , the distance between the ureter and tumor (P = 0.225), or ablation zone (P = 0.089), and total energy (P = 0.739) were included in the multivariate analysis of risk factors. After multivariate logistic regression analysis, only R.E.N.A.L. score (P < 0.001) was related to this complication. The ROC curve is shown in After univariate risk factor analysis, R.E.N.A.L. score (Stenosis of ureter after MWA for the treatment of renal tumor is a rare complication. Rarely had literature reported this complication. Chen et al.  had publOur results showed that the R.E.N.A.L. score is associated with this complication. In contrast to previous research, which concluded the location of the tumor and growth pattern were independent predictors of developing stenosis of ureter . Our resThere are still some limitations to this study—first, the nature of the retrospective study, which might affect the evaluation of outcomes. Second, because the stenosis of the ureter remote from the ablation site after MWA is a rare complication, the series of this complication is still limited; the power of evidence of risk factors is still low. Third, the experience of doctors and single-center study could impact the outcome.In conclusion, R.E.N.A.L. score is associated with ureter stenosis after MWA for the treatment of renal tumor, which combines the information of location, depth, and size of tumor. Preoperative evaluation of the tumor is necessary for avoiding ureter stenosis. Moreover, the combination of various preoperative protective methods might be sufficient to reduce the rate of this complication such as preoperative double-J stent placement, hydrodissection technique, and retrograde cold saline perfusion. In addition, the postoperative remedy is also necessary to decrease the degree of stenosis of the ureter.wjpdabao@gmail.com.The datasets analyzed in this manuscript are not publicly available. Requests to access the datasets should be directed to The studies involving human participants were reviewed and approved by Ethics Committee of PLA General Hospital. Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.JW: protocol and project development, methodology, data collection, manuscript writing, and manuscript editing. JY and ZC: protocol and project development, and methodology. ZH and XY: protocol and project development, and data management. KL and PH: data collection and management. FL and PL: protocol and project development, and manuscript review. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Y. Martin Lo, and Wiley Periodicals, LLC. The withdrawal has been agreed because the article was published in error while it was still under peer review. The article has now been rejected and is withdrawn. The authors bear no fault, and we apologize for the error.The above article, published online on 18 January 2022 in Wiley Online Library (10.1002/fsn3.3087. This duplicate has now been deleted and its DOI redirected to this version of the article.Correction added on 24th November 2022, after initial online publication. A duplicate of this article was published under the DOI:


Anopheles species allowed evolutionary comparisons of the immune response genes in search of alternative vector control of the malarial parasites. Now, with the Anopheles aquasalis genome, it was possible to obtain more information about the evolution of the immune response genes. Anopheles aquasalis has 278 immune genes in 24 families or groups. Comparatively, the American anophelines possess fewer genes than Anopheles gambiaes.s., the most dangerous African vector. The most remarkable differences were found in the pathogen recognition and modulation families like FREPs, CLIP and C-type lectins. Even so, genes related to the modulation of the expression of effectors in response to pathogens and gene families that control the production of reactive oxygen species were more conserved. Overall, the results show a variable pattern of evolution in the immune response genes in the anopheline species. Environmental factors, such as exposure to different pathogens and differences in the microbiota composition, could shape the expression of this group of genes. The results presented here will contribute to a better knowledge of the Neotropical vector and open opportunities for malaria control in the endemic-affected areas of the New World.Anophelines are vectors of malaria, the deadliest disease worldwide transmitted by mosquitoes. The availability of genomic data from various  Annually there are 229 million cases that result in nearly half a million deaths worldwide, affecting mainly pregnant women and children [Anopheles sp. Among more than 400 known species of Anopheles, 41 are recognized vectors worldwide, 9 of which are found in the Americas, including Anopheles aquasalis, the primary malaria vector of coastal Central and South Americas and the Caribbean Islands [Malaria is a severe public health problem in several tropical and subtropical areas. It is caused by parasites of the genus children . The disAnopheles vectors to transmit Plasmodium  are well studied and have been characterized in established African and Asian models such as An. gambiae s. s. and An. stephensi [Plasmodium in the vectors, and three-quarters of the total ingested parasites die inside the midgut at the start of their life cycle [The biological characteristics influenced by variations in the ability of tephensi –5. The dfe cycle . Changesfe cycle ,8.Plasmodium's life cycle in the mosquito [Several families of innate immune genes control the mosquito response to pathogens. They can influence vector competence, allowing or preventing the development of mosquito –11. Alsomosquito ,13.Plasmodium infection and transmission through the synthesis of anti-pathogen molecules, including those expressed after activation of immune signalling pathways. Several components of the IMD (immune deficiency) and TOLL signalling pathways influence the development of murine and human Plasmodium in different anopheline species such as An. gambiae s. s., An. stephensi and An. albimanus [Anopheles Plasmodium-responsive Leucine-Rich Repeat-2 (APL2), and fibrillin-9 (FBN9) [An. stephensi for the continuous expression of REL2 was carried out, allowing the generation of mosquitoes highly resistant to P. falciparum infection [It is assumed that the vector's immune response is predominant in controlling lbimanus ,15. For 9 (FBN9) ,15. Cons9 (FBN9) ,15. Basenfection .An. gambiae s. s. and An. stephensi mosquitoes, proteins dual oxidase:peroxidase and NADPH-oxidase domains (DUOX) and heme peroxidase 15 (HPX15) participate in catalysing the cross-linking of the mucin layer that delimits the food bolus avoiding the direct contact of the intestinal epithelium with the gut microbiota. The silencing of HPX15 increased the expression of the nitric oxide synthase (NOS) protein, the main effector regulated by the JAK/STAT pathway, eliminating both P. berghei and P. falciparum through the TEP1 protein [P. berghei in A. gambiae s. s. [An. stephensi since it does not affect the NOS expression [Likewise, immune mechanisms responsible for microbial homeostasis in the mosquito and molecules related to the structural maintenance of the peritrophic matrix have been described as limiting factors for developing plasmodia ,18. In A protein –21. On tpression .In this sense, pathogen recognition components are so important in the sporogony cycle of the malaria parasite within the mosquito that they have become candidates for the production of transmission-blocking vaccines (TBVs) or genetic modification in the anophelines ,24. As hPlasmodium highlight the importance of understanding the functioning and participation of distinct genes involved in the immune response process. In the future, this knowledge may help raise mechanisms for malaria control. The exciting issue about these data on genetic modification targets or candidates for transmission-blocking vaccines is their conservation among anophelines [Anopheles. Although, in this genomic comparison, intrinsic evolutionary characteristics of the groups or families of genes related to the immune response in anophelines have been recognized. Phylogenetic comparisons could reveal the existence of agonist or antagonist genes against plasmodia in American anophelines, which were detected in African or Asian anophelines in functional experiments [Studies about the interaction between mosquito vectors and phelines ,22,25. Weriments .An. aquasalis to study the composition of the immune response gene families and to make phylogenetic comparisons among four anopheline species of mosquitoes whose genomic data is available. This work is based on homology comparisons with the genes of the immune response of An. gambiae s. s., An. albimanus and An. darlingi, in addition to using the neighbour-joining (NJ) methodology to make evolutionary inferences with the immune response components identified in An. aquasalis.Nevertheless, most of these results were raised from restricted groups of Old World anophelines, which limits what is known about these genes in the neotropical malaria vectors. In the present study, we used the recently annotated genome of the neotropical mosquito . 2. 2.1Anopheles aquasalis mosquito comes from the colony established in the laboratory of Medical Entomology at FIOCRUZ-MG. An. aquasalis protein sequences were obtained from genome annotation (GCA_002846955.1) [An. aquasalis immune response proteins were identified in the genome and annotated proteins by tBLASTn and BLASTp  searches against Anopheles gambiae s. s. immune response proteins available at the ImmunoDB database  [The 46955.1) . Putativon GCA_0046955.1 [et al. [An. aquasalis were characterized with the Conserved Domains Database (CDD) database or Interproscan online tool [An. gambiae s. s. as a control [We used amino acid sequences described by Cao et al.  as modelet al. . The proine tool ,32. Moti control . Other p control ,35.. 2.2An. gambiae s. s. immune response proteins were retrieved from the ImmunoDB. Protein sequences from the new world mosquitoes An. darlingi were also downloaded (Coari AdarC3.8), An. albimanus  and from An. gambiae s. s. (PEST PEPTIDES AgamP4.12) from the VectorBase database website (https://vectorbase.org/) [Culex quinquefasciatus as a species within the Culicidae family, little related to the Anopheles genus, and the An. stephensi mosquito, a species of the Cellia subgenus. Thus, the mean of each family of immune response genes was calculated from the information of the six mosquitoes and the loss or gain of genes was expressed with the standard deviation for each species [se.org/) . To carrse.org/) . The physe.org/) . Finally species .. 3An. aquasalis genome [An. aquasalis genome (BioProject PRJNA389759) has 162 944 Mb, distributed in 16 504 scaffolds (N50 14 431) and according to BUSCO analysis, has a completeness of 96.2% of complete single-copy genes, 0.04% complete and duplicated, 2.3% fragmented and 1.5% missing genes. A total of 12 446 protein-coding genes were predicted in the genome and used for the characterization of the components of the immune response.The data used in this study come from the annotation of the s genome . The assAn. aquasalis genome , unlike An.gambiae s. s., which had approximately 130 more immune response genes. We observed that such differences compared to the An. gambiae s. s. is concentrated in two gene families, fibrinogen-related protein (FREP) and CLIP-domain serine protease (CLIP). While An. gambiae s. s. has 46 FREP proteins, we found only 17 in An. aquasalis. For CLIP proteins, An. gambiae s. s. has 97 annotated proteins, while An. aquasalis has 57 such genes.Two hundred and seventy-eight proteins were identified as of putative immune response function in the Culex quinquefasciatus as an external group, we can observe groups of expansions in An. gambiae s. s. . In New-World anophelines, we observe contractions in leucine-rich immune protein (LRIM), gram-negative binding proteins, prophenoloxidases, C-type lectins and Niemann-Pick C2 proteins (MLD2) compared to An. gambiae s. s. and Cx. quinquefasciatus. Regarding specifically An. aquasalis, our attention is drawn to the loss of peptidoglycan recognition protein, serpins and LRIM proteins, as much as the gain of peroxidase and autophagy-related proteins. In the following sections, we will focus on the results of the classic and conserved pathways and those gene families that showed remarkable gains or losses in An. aquasalis.To facilitate the visualization of gene number variation in each immune-related gene family, we performed a heatmap analysis . The hea. 3.1Plasmodium [An. gambiae s. s. are present in An. aquasalis, An. darlingi and An. albimanus, here an orthologous relationship between each individual member of the family was checked  and the subgenus Cellia (An. gambiae s. s.), a new copy emerged for the African mosquito. A result that was also observed when carrying out the phylogenetic tree with the sequences of 18, STAT2 arises in several of the species of the Cellia subgenus and even in representatives of the Anopheles subgenus. In addition, the absence of STAT2 was not exclusive to American anophelines; the Asian species An. sinensis, does not have a copy of this gene .Among the exciting events to highlight was the duplication of the STAT gene  in his gene . In An. c), An. aquasalis and An. albimanus had orthologues of the well-known JNK cascade proteins shared with An. gambiae s. s., but An. darlingi seems to have lost the PUCKERED gene, a negative regulator of this pathway. It is possible that, in An. darlingi, the PUCKERED gene has its role performed by the dual phosphatase gene since both genes have the same regulatory role in insects such as the fruit fly D. melanogaster [An. aquasalis signaling pathways share identity values with An. gambiae s. s. above 41%. . Within the An. aquasalis annotation, the predictions of the IMD genes, the transcription factor regulator PIAS, TRAF6 , and the NOS genes were fragmented over several scaffolds or missing. Therefore, their predictions were made manually.In the JNK signalling pathway c, An. aqnogaster . Further. 3.2An. aquasalis, different from the 18 genes identified in An. gambiae s. s. The loss of genes was manually reviewed. The SRPN7 and SRPN13 genes from An. gambiae s. s. had a low identity value  in the BLAST alignment against the An. aquasalis genome. Furthermore, this comparison revealed a low score for SRPN7 (102) and low coverage for SRPN13 (12%), making it impossible to annotate these genes manually.A total of 15 genes of the serine protease inhibitor family (serpins – SRPN) were found within the genome of An. albimanus serpins was identified. Most An. aquasalis sequences have values above 52% of identity with An. gambiae s. s.   .Figure . 3.3Besides the proteins from the signalling pathways, superoxide dismutase (SOD) was another group of highly conserved genes in the anophelines. Four genes of this family were identified in all mosquitoes with one-to-one orthologues , with hi. 3.4An. aquasalis six PGRP were identified, a result similar to the finding in other anophelines, where the composition of this family is from five to seven genes [An. aquasalis had identity values above 75% compared to their orthologues in the mosquito An. gambiae s. s. and at least the identity values are above 59.8%; in this group, the orthologue of AgamPGRPS2 in An. aquasalis was fragmented across several scaffolds and was manually annotated. Despite having many single-copy orthologous groups among the four species (LD/LA/S1/LB Groups), in the S2/S3 group, it was impossible to identify orthologues with the AgamPGRPS2 and AgamPGRPS3 proteins for American mosquitoes . In general, there were orthologues one by one in several of the groups identified. However, evolutionary analysis suggests the loss of LRIM-5, 8A, 9, 15 and 26 in An. aquasalis. An. gambiae s. s. was the species that had the most gene expansion in this family, mainly in the subgroup of long proteins (a).The proteins with leucine-rich repeats of the immune response (LRIM) are abundant in anophelines; in escribed a. In genproteins a.Figureb,c; electronic supplementary material, table S1). It was common to find proteins from the two subfamilies between the tree clades without differentiating between the two groups.The LRIM family has been subdivided into four subgroups according to their structural characteristics and previous evolutionary analyzes. They are recognized as lengthy, short, without coiled-coil and transmembrane ,46. The figure 8An. aquasalis, only one member of these proteins with transmembrane parts was identified (group 16). The other anophelines had a more significant number of copies of them, as evidenced in An. gambiae s. s.. These proteins are closely related to the subfamily of long LRIM, with both having a common origin. The long proteins were represented by three groups, composed chiefly of single-copy orthologues for the four anopheline species for two of their genes and the expansion of the APL1 group in An. gambiae s. s. and one copy too much in An. aquasalis was evident (a).Also, the proteins with transmembrane regions (with coiled-coil domains) were represented by groups 15 and 16 in anophelines. In  evident a.b). Likewise, the distribution of each motif can be observed in figure 7c, highlighting that most of the Leu repeat motifs were before the red and yellow motifs, with the conserved residues of Trp and Cys delimiting the LRR domain.As pointed out previously, in the LRIM proteins, the leucine-rich repeats (LRR) domain has a particular distribution pattern of residues essential for identifying some proteins of this group with poor identity values ,47. In tfigure 8. 3.6An. gambiae s. s., up to 46 genes have been identified, and the mosquito An. aquasalis has 17 FREP proteins. Our evolutionary analysis showed four significant expansions in the FREP family in anophelines. The expansions EI and EIII are exclusive of An. gambiae s. s.  genes have heterogeneous abundance in anophelines. In ae s. s. . Convers. 4Cellia and Anopheles subgenera [Anopheles divergence.Conservation of proteins at the amino acid and copy number levels is frequently associated with essential genes for the organism's survival and fitness, considering the classical pathways. At first glance, it is possible to observe that the most conserved gene families (concerning copy numbers) were the immune pathways , with 1 : 1 orthologues in almost all anophelines. Proteins of the IMD, JAK/STAT, TOLL and JNK pathways are among the most studied proteins in anophelines. Their importance in the process of vector/parasite interaction has been demonstrated in many insect vectors. The disruption of any of these main pathways by gene knockout or silencing leads to an inability of the vector to defend against pathogens. Interestingly, in anopheline species, duplication in the STAT transcription factor of the JAK/STAT signalling pathway is recognized for some of the members of the ubgenera ,49. Our An. aquasalis and An. gambiae s. s., SOD3 isoforms have emerged to regulate ROS at specific times and in conditions to combat pathogens in the midgut during blood digestion [Another highly conserved family of proteins identified here relates to the metabolism of reactive oxygen species (ROS) products, as in the case of superoxide dismutase proteins (SOD). This group is universally present and is believed to have arisen long ago in prokaryotes from increased oxygen production in the atmosphere ,51. In Aigestion ,53. The igestion ,54.An. aquasalis and An. gambiae s. s., we observed that some gene families  have similar numbers of genes (in a 1 : 1 orthologous relationships). Among the American anophelines, the only species with the same number of autophagy proteins of An. gambiae s. s. was An. aquasalis. The only exception was the ATG12 gene, which was found in two copies in the An. gambiae s. s. genome [An. aquasalis, An. darlingi and An. albimanus) had only one copy of these genes.By way of the conserved orthologues between . genome , while tArabidopsis thaliana for the ATG12 isoforms and represents a clear example of sub-functionalization. As demonstrated in A. thaliana, where in long periods of starvation and darkness, the silencing of both ATG12B and ATG12A genes resulted in increased mortality. According to the results, any of the two ATG12 is sufficient for the association of ATG8 with autophagic bodies. In addition, its expression changes during the life cycle of the plant, with ATG12A frequently expressed in old plants and ATG12B in young A. thaliana tissues [Several components responsible for regulating the catabolic process of autophagy have been conserved in several phyla of eukaryotes such as plants, worms, arthropods and mammals –58. Even tissues .An. darlingi. This protein participates in the final phase of autophagosome formation. In its absence, there is no direction for constructing the pre-autophagosome structure [An. darlingi may be due to the existence of another protein that can activate autophagosome formation, or it is necessary to annotate ATG8 in this mosquito [ATG8 was one of the proteins conserved in all anophelines except for tructure ,58. Thermosquito .An. gambiae s. s.. The emergence of extra copies in An. gambiae s. s. may be related to a greater capacity for regulating the apoptosis process [An. gambiae s. s.. Previously it was shown that galectins underwent an expansion process almost exclusive to mosquitoes, suggesting an influence of hematophagy and exposure to different pathogens for the diversification of this type of recognition receptors in culicids [Interestingly, other proteins, such as caspases and apoptosis inhibitors related to apoptosis processes, have expanded copy numbers in  process –63. Regaculicids ,65.An. gambiae s. s. galectins, there are data on their participation in infections made with Gram-positive bacteria and with P. berghei, GALE6–8 also increased its expression during exposure to the alphavirus ONNV, GALE8 being an important antagonist of the virus, here these galectins are well related in a clade in which An. gambiae s. s. exhibited a duplication precisely with GALE8/9 [An. aquasalis presented a higher number of 1 : 1 orthologues to An. gambiae s. s. than other American anophelines. In summary, by simple parsimony, it is possible to conjecture that An. aquasalis and An. gambiae s. s. present a conserved ancestral catalogue of genes, with unexpected losses in other American anophelines.In  GALE8/9 ,66. Fina GALE8/9 ,19,22,67 GALE8/9 , such asAn. gambiae s. s. and Ae. aegytpi [Plasmodium control since in An. gambiae s. s. PGRPS2 participates in the reduction of P. falciparum and P. berghei [An. aquasalis), C-type lectins, MLD2 and scavenger receptors. Interestingly, for the last two, their functions are not limited exclusively to the immune response, and their diversification could be due to each organism's metabolic or sensory needs [An. gambiae s. s. and An. stephensi MLD silencing increases the oocyst number of P. falciparum and P. berghei in the midgut, reinforcing the hypothesis previously raised that MLD possibly prevents sterol uptake by parasites [Pattern recognition proteins (PRPs) underwent gene gain and loss dynamics ranging from conserved copy number clusters to extensive species-specific diversifications. For example, in the Gram-negative binding protein (GNBP) and PGRP families, conservation was observed for a few orthologues with possible losses in American mosquitoes. Genes identified as PGRPS2 and GNBP2/3 could have implications in limiting the growth of populations of exogenous bacteria functionally recognized in  aegytpi ,69. The  berghei . Some PRry needs ,72. Alsoarasites ,74.Aedes and Culex mosquitoes, and they are specific for each virus or even for serotypes [An. gambiae s. s. with P. berghei in the proteins CTL4 and CTLMA2, these two lectins type C protect the malarial ookinetes from melanization [An. albimanus, the protective action of these proteins against plasmodia does not exist, and, according to our analysis based on the phylogenetic data, it is suggested that this functional difference is related to the lack of a true orthologue of CTL4 by the American species [As in other insects, CTL proteins (C-type lectins with a single CRD domain) have the most significant diversification in anophelines. In other mosquitoes, it is known that these proteins participate in the homeostatic balance of the microbiota, preventing the activation of antimicrobial peptides (AMPs) . The diverotypes –78. The erotypes . This agnization ,81. Howe species .An. gambiae s. s.. This phenomenon was highly accentuated in the FREP family, a previously recognized group with rapid divergence and relaxed evolutionary constraints in insect lineages such as Diptera [Plasmodium [Drosophila, serving as opsonization molecules to control microbes and pathogens, triggering lysis, phagocytosis or melanization [An. gambiae s. s. concerning proteins related to TEP1, which is associated with controlling the microbiota and the fight against pathogens derived from blood meal [An. gambiae s. s., but not in An. albimanus [An. aquasalis, only one LRR was recognized under the same conditions. However, most of the An. gambiae s. s. LRIMs were present in the New World mosquitoes being 17 of 27 clades composed of single-copy orthologues. Characteristically, Toll receptors showed the same division of three subgroups identified in the insect genomic comparison [P. falciparum in An. gambiae s. s. and has orthologues in the other anopheline species [An. aquasalis and An. albimanus, transcriptomic studies indicate little activity of TOLL receptors against Plasmodium after ingesting a blood meal, even if the TOLL signalling pathway is activated [The species-specific diversifications were much more abrupt in the PRP of the FREP, TEP, TOLL and LRIM families in the four studied anopheline species, especially in  Diptera . FREPs aasmodium . TEP is nization ,85. Therood meal ,86. Inteood meal . LRIMs albimanus ,47,87. Imparison . The div species –90. In Activated ,91. ThusAn. gambiae s. s. in subfamilies B and E [An. darlingi and An. albimanus. In mosquitoes such as Ae. aegypti and An. gambiae s. s., it is widely studied how serine protease with CLIP domains actively regulates melanization as a defence mechanism against bacteria, fungi and parasites [An. gambiae s. s., non-catalytic CLIPs are recruited with the TEP1/LRIM1/APL1C complex. This process mediates the accumulation of the complement system on the surface of microorganisms [Ae. aegypti, for example, CLIP29 and CLIP30/31 paralogues activate the TOLL signalling pathway along with the SERPIN2 protein acting against the fungus Beauveria bassiana [A few authors have asserted that expansions in the families related to the modulation process of the immune response are widespread ,64,92. I B and E , also obarasites ,93–96. Irganisms ,96. Alsorganisms ,98. Somerganisms . SpeciesAn. aquasalis. The PPO-6 clade is associated with the melanization of plasmodia in refractory strains of An. gambiae s. s. and An. stephensi [P. berghei in An. gambiae s. s. by the activation of the PGE2 protein. Moreover, in An. dirus, PPO4 has an antispasmodic effect independent of the melanization [The prophenoloxidase (PPO) system is a hemolymph-based complex of enzymes that, when activated, generate peptides and adhesive proteins, mediating many of the defence functions in arthropods. The expansion of PPO in mosquitoes is well-known. There is a discussion that a series of duplication events have led to the current number of proteins in the culicids ,99,100. tephensi ,99. Howenization ,102.An. aquasalis, the primary malaria vector of coastal Central and South America and the Caribbean Islands. The An. aquasalis mosquitoes used in this study are from a well-established colony that has also analysed the interaction process with murine and human malarial Plasmodium strains. The results raised here will contribute knowledge of the study of a Neotropical vector, which may open opportunities for future malaria control in the malaria-endemic areas of the New World.Overall, the results presented in this study indicate a variable pattern of evolution in the immune response genes of the studied anopheline mosquitoes. Environmental factors, such as exposure to different pathogens and differences in the composition of the microbiota, could shape the expression of this group of genes. This study formulated a catalog of the evolution of immune response genes in


Borrelia burgdorferi sensu lato complex. The disease is characterized by a varied course; therefore, the basis for diagnosis is laboratory methods. Currently, a two-tiered serological test is recommended, using an ELISA as a screening test and a Western blot as a confirmatory test. This approach was introduced due to the relatively high number of false-positive results obtained when using an ELISA alone. However, even this approach has not entirely solved the problem of false-positive results caused by cross-reactive antibodies. Many highly immunogenic B. burgdorferi s.l. proteins are recognized nonspecifically by antibodies directed against other pathogens. This also applies to antigens, such as OspC, BmpA, VlsE, and FlaB, i.e., those commonly used in serodiagnostic assays. Cross-reactions can be caused by both bacterial  and viral  infections. Additionally, a rheumatoid factor has also been shown to nonspecifically recognize B. burgdorferi s.l. proteins, resulting in false-positive results. Therefore, it is necessary to carefully interpret the results of serodiagnostic tests so as to avoid overdiagnosis of Lyme disease, which causes unnecessary implementations of strong antibiotic therapies and delays in the correct diagnosis.Lyme disease is a tick-borne disease caused by spirochetes belonging to the  Borrelia burgdorferi sensu lato complex (B. burgdorferi s.l.). Currently, on the basis of phylogenetic similarity, there are about 21 genospecies within the B. burgdorferi s.l complex; however, due to the attempts made to identify and describe new strains, it is believed that this number is not final [Lyme disease is a tick-borne disease caused by bacteria included in the Borrelia afzelii, Borrelia garinii, Borrelia bavariensis, Borrelia burgdorferi sensu stricto (s.s.), Borrelia lusitaniae and Borrelia spielmanii. For many years, it was believed that Borrelia burgdorferi s.s. was the only causative genospecies of Lyme disease in North America. However, in recent years there have been more and more reports suggesting that Borrelia bisettii and Borrelia mayonii may also cause Lyme disease in the USA and Canada [Not all bacteria belonging to this group cause borreliosis. In Europe, there are six genospecies causing the disease in humans, i.e., d Canada ,3,4.B. burgdorferi s.l. are ticks of the genus Ixodes, and it is the area of their natural existence that determines the zone of Lyme disease occurrence. There are four main species of ticks that transmit Lyme disease to humans: Ixodes scapularis in the eastern United States and Canada; Ixodes pacificus in the western United States; Ixodes ricinus in Europe and Asia; and Ixodes persulcatus found only in Asia [B. burgdorferi s.l. when feeding on vertebrates that are reservoirs for pathogens. After B. burgdorferi s.l. enters the arachnid’s body with blood, the spirochetes localize in its midgut. The increase in temperature and the change in pH inside the tick, accompanying the blood collection, are stimuli for B. burgdorferi s.l. to start expressing a new gene pool. This allows the spirochetes to migrate from the midgut to the tick’s salivary glands, from where they enter the body of a new vertebrate host (including humans) together with the saliva [The vectors transmitting  in Asia . Ticks be saliva ,7. Lyme disease was first described in the population of Lyme, Connecticut, USA. Rheumatoid-like arthritis was noted in them, which mainly affected children . HoweverB. burgdorferi s.l. does not produce classic toxins or other recognizable virulence factors that directly cause damage to the host tissues. Therefore, it seems that multisystemic disorders are caused by too strong immune reactions to bacterial components [mponents . In the mponents ,14,15. B. burgdorferi s.l. about 3–30 days after the tick bite [B. burgdorferi s.l. representatives [B. afzelii is associated almost exclusively with acrodermatitis chronica atrophicans (ACA). B. garinii and B. bavariensis are often associated with neurological symptoms, and B. burgdorferi s.s. mainly causes Lyme arthritis (LA) [The most characteristic manifestation of early Lyme disease is a skin lesion called erythema migrans (EM), which appears in about 60–90% of those infected with ick bite ,16. At tick bite ,18. If tick bite ,15. One ick bite ,19,20. Tntatives . Therefotis (LA) ,23,24.B. burgdorferi s.l. in clinical samples and the difficulties in their cultivation, indirect tests based on the detection of class M and G antibodies are most often used [B. burgdorferi s.l. proteins used in them and nonspecific antibodies contained in human sera, make diagnostics difficult, leading to overdiagnosis of Lyme disease. This has serious implications for patients who are then subjected to unnecessary strong antibiotic therapy, and the correct diagnosis is delayed, which can lead to catastrophic consequences.Due to the fact that the symptoms of Lyme disease are not specific enough to allow for a correct diagnosis, laboratory assays play an important role here. As direct methods have many limitations connected with the presence of a very low number of ten used . UnfortuThis article presents the problem of cross-reactions in Lyme disease serodiagnostics leading to false-positive results and discusses possible solutions to this problem.B. burgdorferi s.l. are microareophilic, Gram-negative bacteria belonging to the type Spirochaetes [B. burgdorferi s.l. cell: the outer lipid membrane with many surface lipoproteins, the periplasmic space, the flagellum and the inner cytoplasmic membrane forming the protoplasmic cylinder. The flagella and peptidoglycan layer are located in the periplasmic space [The ochaetes ,27. Due ochaetes ,29. The ic space ,29.B. burgdorferi s.l. consists of a linear chromosome and numerous circular and linear plasmids. The chromosome mainly encodes the housekeeping genes and its sequence is quite strongly conserved within B. burgdorferi s.l. Plasmids carry genetic information responsible for the virulence and pathogenicity [B. burgdorferi s.l. isolates have a complete set of them. Only the cp26, cp32 and lp54 plasmids are necessary for the survival of the bacteria in the environment; therefore, they are present in all B. burgdorferi s.l. isolates and show a relatively high degree of conservation of the nucleotide sequence. The rest of them have low conservation and are quite easily lost, which is associated with a great difficulty in the laboratory cultivation of fully virulent strains of B. burgdorferi s.l. [The complete genome of genicity . So far,eri s.l. ,31,32. B. burgdorferi s.l. are also characterized by a very diverse proteome. The bacteria have the ability to change their surface antigens as an expression of adaptation to changing environmental conditions. Different proteins are present on the surface of the pathogen’s cell during its existence in ticks and vertebrates. The production of proteins necessary for mammalian entry and colonization usually begins in response to changing conditions during a tick’s blood meal. An increase in temperature and a decrease in pH are a signal to start the migration of spirochetes to the salivary glands of arachnids and the production of new proteins needed for transmission [Due to the complicated structure of the genome, smission ,33.Borrelia proteins, the best known are surface antigens, which fall into two main categories: lipoproteins, which are anchored to the outer membrane by lipid moieties at the N-terminus, and integral outer membrane proteins (OMPs), which are anchored by transmembrane domains [B. burgdorferi s.l., which is influenced by the location of almost all known genes encoding OMPs on the chromosome. They are mainly responsible for functions necessary for the survival of the bacterial cell, such as obtaining nutrients or resistance to antibiotics [B. burgdorferi s.l. It is mainly the expression pattern of plasmid-encoded lipoproteins that changes throughout the life cycle of the spirochete in response to changes in environmental conditions. This indicates that surface lipoproteins play an important role in virulence and host–pathogen interactions, and they have also been shown to be involved in host immune response evasion. This opinion is supported by the observation that the loss of plasmids correlates with the decrease in B. burgdorferi s.l. infectivity [Among all  domains . OMPs aribiotics ,35. Lipoectivity ,36,37.B. burgdorferi s.l. proteins is provided in A brief description of the most immunogenic B. burgdorferi s.l. in the collected samples and the long waiting time for the result. The above-mentioned limitations mean that this method is mainly used in the case of patients with dysfunctions of the immune system [For most bacterial diseases, the direct detection of pathogens via culture is considered the gold standard. Unfortunately, this approach is not used in the diagnosis of Lyme disease due to the very high growth requirements of the spirochetes, the low number of e system ,63,64.B. burgdorferi s.l. were selected in the WB test. It has been established that for the detection of immunoglobulin M (IgM) the most useful antigens are OspC (p23–p25), BmpA (p39) and FlaB (p41), and the test is interpreted as positive when there is a reaction to at least two of them [B. burgdorferi s.l. are the main source of antigens in both methods.Currently, a two-tiered serodiagnostic approach is recommended in the diagnosis of Lyme disease. Due to their high sensitivity and low cost, ELISA tests act as screening tests. However, a positive result does not mean a reliable diagnosis of Lyme disease, as ELISA tests have a fairly low specificity. The Western blot (WB) is used as a second-stage confirmatory test, because, due to the ability to distinguish individual protein bands, it is much more specific. In order to achieve the best diagnostic utility, the most reactive and characteristic antigens produced by  of them . However of them . Whole-cHowever, the serodiagnosis of Lyme disease affects several problems; one of them is the serologic window—the time needed for the immune system to produce specific antibodies against pathogens during primary infection. IgMs appear first in the host organism; it takes place about 2 weeks after infection, while the production of IgG usually begins after 3–6 weeks, and they replace IgM as the disease progresses. As the infection continues, the immune response gradually matures so that the sensitivity of serological tests during the later stages of the disease increases significantly ,67. B. burgdorferi s.l., and the low degree of preservation of amino acid sequences of proteins among them also have a negative impact on the effectiveness of Lyme disease serodiagnostics. This problem is particularly relevant in Europe, where there are as many as six genospecies pathogenic for humans. Most of the immunogenic proteins are heterogeneous, and thus the use of WCL with only one genospecies as antigen preparations in serodiagnosis carries the risk of obtaining a false-negative result [B. burgdorferi s.l. It is also problematic to obtain WCL containing all of the most immunogenic antigens, as some of them are produced only in vivo [B. burgdorferi s.l. produces many proteins homologous among microorganisms, which carries the risk of cross-reactions and leads to false-positive results in serodiagnostic tests. The lack of specificity of diagnostic tests for Lyme disease prompted the introduction of a two-tiered testing strategy in 1995 [Another limitation is the great genospecies diversity of e result ,32,68,69 in vivo ,70. The  in vivo ,71,72. A in 1995 ,74.B. burgdorferi s.l. genospecies that caused the infection, which will simplify the interpretation of diagnostic assays. Recombinant proteins are already used in many commercially available serodiagnostic WB and ELISA tests , recomLine Borrelia IgG/IgM (Microgen), Anti-Borrelia plus VlsE (Euroimmun), the ZEUS ELISA Borrelia VlsE1/pepC10 IgG/IgM Test System  and the C6 Lyme ELISA kit  and displace native B. burgdorferi s.l. proteins. The results obtained by the researchers indicate that recombinant proteins or synthetic peptides have the potential to solve some problems affecting the diagnosis of Lyme disease. Careful and thoughtful antigen selection can reduce cross-reactivity and allow test sensitivity to be independent of the B. burgdorferi s.l. complex, they are often added in variants from different genospecies for serodiagnostic assays. Thanks to this, it is possible to detect diseases caused by different representatives of B. burgdorferi s.l. with one test. C6 is a 25 amino acid peptide derived from VlsE. Its sequence is conserved among B. burgdorferi s.l., and therefore C6 is widely used in immunodiagnostic assays [The most widely used in commercial diagnostic tests are DbpA, OspC, VlsE and synthetic peptides C6 and pepC10. DbpA and VlsE proteins are mainly used in the detection of IgG antibodies. Due to the high variability of DbpA within the c assays . OspC anc assays . The specificity of the two-level serological tests in research studies exceeded 99%; however, under these conditions they were performed by highly efficient reference laboratories. Many reports indicate that in real clinical practice two-tiered tests show a much lower specificity ,74Although there are many guides that accurately describe the symptoms of Lyme disease at its various stages and the diagnostic tests that ought to be implemented, these recommendations are often ignored. This results in ordering tests for Lyme disease in cases where there are no clear medical indications for it, which increases the rate of falsely diagnosed Lyme patients. Therefore, serodiagnostic testing should be avoided in individuals who are not at risk of being bitten by a tick or who have only very general symptoms  ,77. TherThis is especially true for IgM detection as these are first-line antibodies. Therefore, their presence is often associated with the acute phase of infection. Nevertheless, their reactivity is often nonspecific because IgM is produced when the immune response to the foreign antigen is still immature. IgM should therefore only be used in cases of suspicion of early Lyme disease, i.e., in cases of NB lasting <6 weeks or Lyme carditis (LC). Other clinical symptoms, like LA and ACA, are manifestations of late borreliosis and appear after at least 6 weeks. Therefore, in these cases tests focused on the detection of more specific IgG should be carried out .B. burgdorferi s.l. Studies have shown that in some regions of Europe in high-risk groups  the prevalence of antibodies against B. burgdorferi s.l. antigens is even over 50%, which means that a positive test result does not always mean that the disease is active [The usefulness of serological tests is also influenced by the high prevalence of anti-B. burgdorferi s.l. antibodies. This is especially problematic in Europe, as according to a 1995 WHO report the whole continent ought to be considered the endemic area of s active ,80,81.B. burgdorferi s.l. antigens that were mainly recognized by specific IgM many years after infection were OspC and FlaB. This means that antibodies to proteins such as DbpA (p18), BmpA (p39), p45, p58 and p66 have disappeared over time [It also turns out that false-positive results may be due to the long duration of the anti-Borrelia antibody in the body. Kalish et al. (2001) studied 79 patients who had been diagnosed with Lyme disease 10–20 years earlier and now showed no signs of active infection. Using a two-tiered test, it was shown that as many as 25% of them had specific IgG and 10% had specific IgM . The B. ver time . Howeverver time .There are several infections/diseases that can be misdiagnosed as Lyme disease or vice versa. The most problematic are syphilis, relapsing fever and viral infections caused by Epstein–Barr virus (EBV) and cytomegalovirus (CMV). Furthermore, people positive for rheumatoid factor (RF) are also often misdiagnosed with borreliosis ,85,86,87B. burgdorferi s.l. This group includes such species as Borrelia hermsii, Borrelia miyamotoi, Borrelia hispanicaBorrelia duttonii and Borrelia recurrentis. These bacteria can be divided into lice-borne fever Borrelia (LBRF) and tick-borne relapsing fever Borrelia (TBRF). TBRF Borrelia are typically transmitted by soft ticks of the genus Ornithodoros. Currently, it seems that only Borrelia miyamotoi is transmitted by hard-bodied ticks; likewise, B. burgdorferi s.l. LBRF is caused only by Borrelia recurrentis [Borrelia may be found on all continents except Australia and Antarctica and is a serious public health problem in some parts of the world. LBRF is reported mostly in Africa and single cases in Europe occur mostly among refugees. During the development of relapsing fever, many nonspecific symptoms  and neurological disorders  may occur. Some of them are similar to the symptoms of Lyme disease, so the clinical differentiation of these two infections is impossible in most cases. It is worth remembering that EM does not appear during relapsing fever, which is one symptom that allows easy and undeniable differentiation of these diseases. In the past, B. hermsii whole-cell lysate was used for relapsing fever diagnostics; however, too much antigen similarity between the two groups of bacteria caused a large number of false-positive results. Nowadays, qPCR is mainly used to diagnose relapsing fever. Also, serological tests based on the glycerophosphodiester phosphodiesterase (GlpQ), absent from B. burgdorferi s.l., have been developed [Relapsing fever Borrelia (RFB) are the bacteria closely related to urrentis . TBRF Boeveloped . Treponema pallidum. Serological tests for syphilis at all stages of infection remain the mainstay of diagnosis. Antibodies against T. pallidum antigens often react with B. burgdorferi s.l. proteins, particularly with ELISAs when WCL is used as the source of antigens. This is due to the close phylogenetic relationship of the two pathogens; this cross-reactivity is strongly related to shared flagellar antigens [B. burgdorferi s.l. proteins, such as OspC, BBA64, BmpA, FlaB, OspF and OspC, BBA64, BmpA, FlaB and OspF in IgG [B. burgdorferi s.l. does not appear to lead to a false-positive syphilis test [B. burgdorferi s.l. and T. pallidum can be minimized by incubating the sera with Reiter treponema preparations. However, this procedure did not eliminate all nonspecific interactions [Syphilis is a sexually transmitted disease caused by spirochetes antigens . In the F in IgG . So, it lis test . It has ractions ,92.Anaplasma phagocytophilum, Gram-negative, intracellular bacteria. The main route of transmission of A. phagocytophilum is a bite of Ixodes ticks. Despite the apparently ubiquitous presence of A. phagocytophilum in ticks and animal reservoirs, confirmed clinical cases of HGA in Europe are rare compared to the rest of the world. It has been shown that sera collected from HGA patients show cross-reactions with B. burgdorferi s.l. proteins, such as OspC, BBA64, p37, FlaB, VlsE, OspA, OspC and OspF in the M class of antibodies and OspC, p37, FlaB, OspA and OspF in the IgG. Interestingly, despite the detection of these nonspecific antigen–antibody reactions, ELISA tests performed on the WCL did not give false-positive results [B. burgdorferi s.l. and A. phagocytophilum antigens is not always due to cross-reactions. People who live in Ixodes tick endemic areas and who experience multiple tick bites are probably exposed to the transmission of different pathogens. It is possible that a patient with an HGE infection was previous exposed to B. burgdorferi s.l. or vice versa and may produce antibodies to both pathogens. Furthermore, co-infections in humans and vectors have been confirmed [Human granulocytic anaplasmosis (HGA), formerly known as human granulocytic ehrlichiosis, is caused by  results . Howeveronfirmed ,94. Theronfirmed ,95.Yersinia enterocolitica and Yersinia pseudotuberculosis by ingesting contaminated food or water or from direct infection through blood transfusions. The infection may be asymptomatic, but, in some cases, reactive arthritis similar to LA caused by B. burgdorferi s.l. may develop. The most commonly affected joints are the knees and ankles; but other joints, such as those of the toes, fingers, and wrists, may be involved. As in the case of Lyme disease, the diagnosis via cultures is not very sensitive; therefore, serological tests detecting antibodies specific to Yersinia antigens may be helpful [Yersinia outer proteins (YOPs) [B. burgdorferi s.l.-specific antibodies and the YOPs in Western blots. For anti-Borrelia IgG, cross-reaction was detected with YopH, YopB, V-ag, YopD, YopN, YopP and YopE, and it was detected for IgA with YopD [Borrelia serum samples with this observed cross-reactivity contained IgG against FlaB and specific IgG and IgM against OspC. It may prove that there is antigenic similarity between OspC and FlaB antigens of B. burgdorferi s.l. and YopD of Yersinia and that two-way cross-reactivity is present. It has been shown that p60, FlaB, OspA and OspC B. burgdorferi s.l. antigens are highly cross-reactive with anti-Yersinia sera [People get infected with  helpful ,97. The s (YOPs) . Cross-rith YopD ,97. All nia sera ,97. In tB. burgdorferi s.l. In the studies conducted by Panelius et al. (2002), the percentage of patients with a positive EBV result in the OspC ELISA was initially as high as 73%; however, this cross-reactivity was limited by the addition of sodium thiocyanate and dropped to 46%. Unfortunately, the effect of decreasing nonspecific reactions was not seen with sera specific for syphilis and rheumatoid factor [Epstein–Barr virus is a gamma herpesvirus causing infection in humans worldwide. The prevalence of EBV is very high; about 90% of adults are infected with EBV. Most infections occur in young children and are asymptomatic or cause nonspecific symptoms . It was d factor .Herpesvirales. Usually, CMV infection is asymptomatic, but in people whose immune system is defective or immature, such as newborns, people with an overgrowth and patients with AIDS, it can be a serious problem. The virus is highly distributed and the worldwide prevalence of CMV has been estimated at 83% (66–90%) [B. burgdorferi s.l. antigens [B. burgdorferi s.l. WCL showed a specificity of only 60% for the detection of antibodies in class M. The low specificity in tests with the use of CMV sera was due to the cross-reactivity of IgM with the OspC and FlaB (p41). In addition, it was shown that anti-CMV IgG recognized such B. burgdorferi s.l. antigens as OspC, FlaB, BmpA and VlsE [Cytomegalovirus belongs to viruses in the order (66–90%) . This meantigens ,101. Somand VlsE .B. burgdorferi s.l. [B. burgdorferi s.l and other unrelated pathogens, such as Salmonella and Campylobacter. These antibodies can persist in the circulation for up to 3 months. False-positive results for Lyme disease were obtained not only in the EIA tests but also in the more specific Western blot. Therefore, there is a possibility that an acute B19V infection could be misinterpreted as Lyme disease [Parvovirus B19 (B19V) is a small, single-stranded, nonenveloped DNA virus. In adults, it can cause joint pain or arthritis with pronounced morning stiffness that may imitate LA caused by eri s.l. . Acute B disease . One of  disease . B. burgdorferi s.l. antigens, including LA7 (p22), BBA64 (p35), p37, BmpA, OppA2, VlsE and OspC [One of the most common symptoms of late borreliosis  is Lyme arthritis. LA shares common clinical features and a synovial histology with rheumatoid arthritis (RA), which means the clinical symptoms are often indistinguishable. Recommended serological tests might also lead to misdiagnosis due to the presence of RF, which is present in the serum of patients with arthritis  and otheand OspC ,106,107.and OspC .B. burgdorferi s.l. antigens, like p37, BmpA, OspF and OspC, occur; however, this phenomenon has not been fully understood [Additionally, it was observed that in patients with oral infections cross-reactions with derstood ,108. B. burgdorferi s.l. antigens, trying to increase the specificity of diagnostic tests [The problem of cross-reactions in the serodiagnosis of Lyme disease has been known for a long time. In the early 1990s, a study was carried out to identify cross-reactive ic tests . ResearcB. burgdorferi s.l. during their persistence in tick organisms. Nevertheless, numerous studies have demonstrated the presence of antibodies against these proteins in the sera of individuals afflicted with Lyme disease. OspA has even been included as a specific band in the WB reference assays [A. phagocytophilum, T. pallidum, Yersinia, E. coli and B. hermsii may unspecifically bind OspA and/or OspB [OspA and OspB are among the proteins that can lead to false-positive results in serodiagnostic tests for Lyme disease. This is rather surprising, considering that they are produced by e assays ,66,85. I/or OspB . Additio/or OspB . Another/or OspB . It has /or OspB .B. burgdorferi s.l., it has been shown that individuals with various infections and disorders may develop antibodies that nonspecifically bind to the OspC protein [B. burgdorferi s.l. and is maintained at a high level by continuous stimulation with cross-reactive autoantigens or with antigens from other microorganisms or environmental factors. Diet can also contribute to this, as the PKKP motif can be found in the glutathione transferase of important crops, such as common wheat and barley [Another protein with well-described cross-activity is OspC. Although widely applied as an antigen in commercial diagnostic assays for detecting both IgG- and IgM-specific antibodies of  protein . These d protein . Moreoved barley .Treponema spp. Especially conserved among these bacteria are the fragments at the ends of the protein ; the central part of the antigen shows higher variability [FlaB p41) shows a high sequence identity of about 40% with flagellin proteins of iability . FlaB sh shows a iability . Howeveriability .B. miyamotoi variable major proteins (Vmps) belonging to the variable large protein (Vlp) family, which are VlsE homologues. B. miyamotoi is the etiological factor of TBRF, transmitted by Ixodes ticks and likewise B. burgdorferi s.l. Cross-reactions of anti-B. miyamotoi antibodies against the C6 peptide were confirmed in a study with experimentally infected mice [B. miyamotoi and B. burgdorferi s.l. occupy the same endemic region, so it should be noted that the C6 peptide alone does not allow serological differentiation between the two tick-borne diseases. Therefore, in order to distinguish borreliosis from B. miyamotoi infection, a two-stage examination is necessary, along with a critical evaluation of the clinical symptoms [In silico analyses showed that the C6 peptide was similar to domains within the ted mice . This crsymptoms .Chandra et al. (2011) reported that about 20% of healthy individuals with no history of Lyme disease had IgG antibodies to p66 , which iB. burgdorferi s.l. antigens, such as BmpA, BBA64, OspF, LA7 and OppA2, has also been proven  are nonspecific, making it impossible to diagnose it properly on the grounds of the clinical picture. For this reason, laboratory methods are the basis for Lyme disease diagnosis, where detection of specific IgM and IgG is the most common . The serB. burgdorferi s.l. proteins [T. pallidum [Yersinia [It is these nonspecific interactions that impose the currently recommended two-tiered algorithm in the serodiagnosis of Lyme disease, increasing the costs of routine diagnosis of Lyme disease and extending the time necessary to make a final diagnosis. It seems that the most problematic in the current approach is the WB, which is time-consuming and difficult to automate. In addition, its reading, as already mentioned, is subjective, which leads to significant discrepancies between laboratories ,77,78,79proteins ,109,123.proteins  and T. ppallidum ,90 and YYersinia ,97,124 bKnowledge about cross-reactivity is mainly obtained as complementary information when determining the diagnostic utility of new tests based on recombinant proteins ,86,108. It is therefore necessary to conduct comprehensive, wide-ranging research using uniform standards. This will allow for the clear identification of infections/disorders and antigens that may potentially cause cross-reactions in the serological diagnosis of Lyme disease, which will allow diagnosticians to be better prepared to interpret the results. Additionally, it is necessary to follow official recommendations during the diagnostic process. It is important to remember what symptoms are the manifestation of late borreliosis and then perform only detection of highly specific IgG.B. burgdorferi s.l. proteins, a specific one-stage assay superior to the standard two-tiered assay has not yet been developed [B. burgdorferi s.l. and not cross-react with immunoglobulins specific for other antigens.Moreover, the creation of new diagnostic tools may contribute to solving this problem. It has been shown that the use of selected recombinant proteins instead of WCL in serodiagnostic tests can significantly reduce the rate of false-positive results ,108. Deseveloped ,78,79. Eeveloped ,126. Howeveloped ,128. ThiB. burgdorferi s.l. [B. burgdorferi s.l. and eliminate those commonly found in antigens of other pathogens. Already, in the example of VlsE and FlaB, it has been shown that the proper selection of protein fragments allows for increasing the specificity without reducing the sensitivity of the immunoassay [The easiest and most widely available approach for designing new specific antigens for the diagnosis of Lyme disease is the analysis of amino acid sequences of proteins. Currently, there are a large number of databases containing the amino acid sequences of proteins from many organisms. The most popular are UniProt  and NCBIeri s.l. ,132 protunoassay ,134.Even greater possibilities in the selection of fragments for the construction of chimeric proteins are provided by B-cell epitope mapping, which allows for detailed knowledge of the distribution of immunodominant fragments and potential cross-reaction fragments ,136. TheComputational methods predict the existence of potential epitopes based on such physicochemical properties as hydrophilicity, solvent accessibility, flexibility, turns, polarity, antigenicity and surface exposure. In general, computational epitope prediction methods can be divided into two groups depending on whether the input is only the amino acid sequence of the antigen or its tertiary structure. These methods using the three-dimensional structure of the protein are characterized by a greater regularity; unfortunately, it is not always possible to use them due to the fact that the spatial structure of many proteins is not known . HoweverThe information provided by empirical techniques is much more complete and reliable. Experimental B-cell epitope mappings involve methods such as peptide microarray , X-ray c


An 8-year-old previously healthy female was initially found to have a small pit on her nasal dorsum. The lesion developed local infection and she was initially treated with antibiotics 2 years prior to the current presentation. The lesion was diagnosed as a dermal sinus tract, and surgical removal was conducted at an outside hospital. In retrospect, the preoperative work-up imaging showed an occult intraosseous nasal bone extension; however, this was not appreciated at the initial surgery. She experienced repeat infections and underwent a second surgery with exploration under the nasal bones; however, the patient experienced recurrent post-operative local infections. The family presented to our institution for a second opinion.On review, the head CT demonstrated an intraosseous cystic lesion completely hidden in the nasal bone Fig. . The lesTo date, no previous reports have documented a case with intraosseous dermoid cyst which was completely hidden in the nasal bone. An extension of the dermoid cyst below the nasal bone has been identified in 10% of patients in a large series of nasal dermoid cysts  previousWritten informed consent was obtained from the parent of the patient for publication of the details of their medical case and any accompanying images. This study protocol was reviewed and the need for approval was waived by the Boston Children's Hospital Institutional Review Board.The authors have no conflicts of interest to declare.The authors received no financial support for the research, authorship, and publication of this article.Design of the manuscript: Yusuke S. Hori, John G Meara, and Mark R. Proctor; acquisition, analysis, and interpretation of data: Yusuke S. Hori, John S. Albanese, John G. Meara, and Mark R. Proctor; drafting the manuscript: Yusuke S. Hori; and revising the manuscript: Yusuke S. Hori, John G. Meara, and Mark R. Proctor.All data generated or analyzed during this study are included in this article. Further inquiries can be directed to the corresponding author.


Rhipicephalus sanguineus sensu stricto and Rhipicephalus linnaei has been confirmed. Both species are found in sympatry in the southern United States, northern Mexico, southern Brazil, and Argentina. The objective of this work is to evaluate the projection of the potential distribution of the ecological niche of Rhipicephalus sanguineus sensu lato in two climate change scenarios in Mexico and the border with Central America and the United States. Initially, a database of personal collections of the authors, GBIF, Institute of Epidemiological Diagnosis and Reference, and scientific articles was built. The ENMs were projected for the current period and two future scenarios: RCP and SSP used for the kuenm R package, the ecological niche of R. sanguineus s.l. It is distributed throughout the Mexico and Texas (United States), along with the border areas between Central America, Mexico, and the United States. Finally, it is observed that the ecological niche of R. sanguineus s.l. in the current period coincides in three degrees with the routes of human migration. Based on this information, and mainly on the flow of migrants from Central America to the United States, the risk of a greater gene flow in this area increases, so the risk relating to this border is a latent point that must be analyzed.In America, the presence of  Rhipicephalus sanguineus complex) of Afrotropical and Palearctic origin . Unlike mechanistic models, correlative models use global variables (for example: bioclimatic variables based on temperature and precipitation) to explain the potential distribution of a particular species [The most complete study based on the terms of ecological niche construction and model uncertainty was carried out by Alkishe et al. . This ste et al.  used aroe et al.  used 593e et al.  made a fr Mexico . Then, Sr Mexico  used a tand time . We used species ,31.R. sanguineus could increase in areas experiencing warmer and/or longer summers, consequently increasing the risk of transmission of zoonotic agents  [R. sanguineus complex in regions of northern temperate Europe [R. sanguineus s.l. ticks is uncertain. This should not be left aside when we analyze the impact of R. sanguineus s.s. and R. linnaei.Another similarity between both studies  is that kettsii) . On the e Europe . HoweverRhipicephalus for Mexico presents a few useful data [R. sanguineus s.s. and R. linnaei). It is important to note that in previous studies in Mexico using the amplification and sequencing of the mitochondrial 16S-rDNA gene, it has been shown that R. linnaei is the most widely distributed species in the country, described in 10 states of the Neotropical region, in contrast with R. sanguineus s.s. which is restricted to the Nearctic states of Sonora and Chihuahua [R. sanguineus s.s. limits its zone of sympatry with R. sanguineus s.s. in a couple of Mexican states. Since in the present work, only the morphological records prior to the generation of the taxonomic keys of Nava et al. [R. sanguineus in Mexico is projected. This may be due to building the ecological niche with all the variables taking into account this complex species. It is very important to highlight the need to be able to define the limits within the species complex, taking into account ecological, genetic, and morphological issues. An essential point in the construction of the ecological niche is the feasibility of the kuenm R package to be able to establish different sets of variables and to be able to evaluate the best model among all of them. This was one of the limitations of the study by Sánchez-Pérez et al. [R. sanguineus complex based on ecological, genetic and morphological issues but from the point in view of using the same individuals to assess those aspects. Finally, something that has not continued is a review of the areas where the potential distribution exists to find out if the report is true positive or false positive. This information could be useful to validate the ENM used.The ENM of the genus ful data , emphasihihuahua . This isa et al.  and Slapa et al.  are avaia et al. , we havez et al. . In the R. sanguineus s.l. in the Americas, with emphasis on the Southeastern United States [R. sanguineus in Mexico and the border with the United States. When we evaluate the RCP’s and SSP’s scenarios, we can observe that there are areas that overlap between both models, which helps us to have less uncertainty when interpreting the results. In R. sanguineus is scarce, except Tabasco, Chiapas, Yucatan Peninsula and Texas (United States). Currently, comparisons have been made between different general circulation models in different climate change scenarios. However, in this paper, we compare the last two IPCC reports (RCP’s and SSP’s) [R. sanguineus s.l. In Alkishe et al. [R. sanguineus complex. Therefore, it is crucial to be able to have an ecological niche based on morphological, genetic and ecological characteristics and to be able to participate in decision making in different areas.Our results of the ENM of RCP4.5 and RCP8.5 are similar to those proposed by Alkishe et al. . Howeverd SSP’s) . The cone et al. , they maR. sanguineus has yet to occupy its entire potential range for current climatic conditions, although there is already evidence for the tick expanding in range. Previous studies have shown that R. sanguineus s.s. tends to establish itself in temperate regions in Brazil, Argentina, the United States, and Mexico, so in these four countries, the epidemiology of tick-borne pathogens changes radically compared to countries in South and Central America in which only one of these species circulates. For this reason, the niche model is a priority on a regional and/or local scale [Our results are complementary to what was presented by Pascoe et al.  in theiral scale ,4. Our sR. sanguineus s.l from cattle in five municipalities of Chiapas. These ticks were found to be infected with R. rickettsii and Rickettsia felis in the municipalities that correspond to the human migration routes . Therefore, there is an urgent need to carry out active acarological surveillance on the southern border of Mexico. On the Northern border of México, the state of Arizona is a focal point between both countries. Recently, Brophy et al. [R. sanguineus s.s. as a vector of R. massiliae, whereas R. linnaei is a vector of R. rickettsii. For this reason, and given previous findings of both bacterial species across the state of Chihuahua, the outlook for the transmission of a broader range of pathogens increases, for which reason distribution models based on the future are a priority. They will be used to identify risk regions that should be considered for the implementation of differential diagnostic methods for the species of pathogens that both tick specie may be carrying regionally. A clear example is Haemaphysalis longicornis, which transmits the pathogen Theileria orientalis var. Ikeda. This species is native to Asia; however, it has recently invaded the United States and has the potential to invade Mexico [Recently, Danis-Lozano et al.  collectey et al.  found the Mexico .R. sanguineus was found in cattle. Finally, based on this information, and mainly on the flow of migrants from Central America to the United States, the risk of a greater genetic flow in this area increases the risk on this border. This is a latent point that must be analyzed. According to R. sanguineus s.l.; however, these migration routes highlight the latent alarm associated with the displacement of this tick, as it belongs to the migrant caravans that constantly leave Tapachula, Chiapas, looking for its border with the United States.In recent years, the Comisión Nacional de Ayuda a Refugiados has increased the number of migrants who are carrying out their process in the different offices in Mexico. In the year 2021, approximately 110,000 registered migrants were identified; however, the number of unregistered migrants may be higher . ApproxiR. sanguineus s.l. for Mexico and the border of the United States. The information collected is the most recent in terms of the construction of the ecological niche. Our results have a high suitability in the large regions of Mexico, and in different scenarios (RCP’s and SSP’s) the increase is greater. The routes of human migration could be areas of movement of ectoparasites throughout the Mexican territory. Therefore, we sound the alarm regarding the need for control measures for active acarological surveillance on the northern and southern borders of Mexico, in order to resolve the challenges as soon as possible.This article is one of the first to focus mainly on