Scotogramma trifolii Rottemberg, and the spotted clover moth, Protoschinia scutosa (Denis & Schiffermuller), are worldwide polyphagous pests, and the larvae feed mainly on the leaf backs of many agricultural crops. However, the food sources and feeding habits of the adults are still poorly known. We investigated the ultramorphology of the proboscis and associated sensilla of S. trifolii and P. scutosa using scanning electron microscopy. The results show that the proboscises of S. trifolii and P. scutosa are structurally similar, both including three sensillum types and three zones (Zone 1–3). The sensillum chaeticum is non-porous hair-like, the sensillum basiconicum is a short smooth cone with a sensory pore on the blunt tip, and each sensillum styloconicum is composed of a uniporous sensory cone inserted into a ribbed stylus. In addition, the movement and fluid uptake mechanisms of the proboscis and the possible function of sensilla are briefly discussed.The clover cutworm, Scotogramma trifolii Rottemberg, and the spotted clover moth, Protoschinia scutosa (Denis & Schiffermuller), are serious polyphagous pests, attacking numerous vegetables and crops, resulting in huge economic losses. However, the feeding behavior and mechanisms of the adult stage remain unsatisfactorily explored. In this study, the proboscis morphology of S. trifolii and P. scutosa are described in detail using scanning electron microscopy, with the aim of investigating the morphological differences and feeding behavior of these two species. The proboscises of S. trifolii and P. scutosa are similar in morphology and structure and are divided into three zones (Zone 1–3) based on the morphological changes of the dorsal legulae. Three sensillum types are located on the proboscises of both species, sensilla chaetica, sensilla basiconica, and sensilla styloconica. Significant differences were observed in the length of the proboscis and each zone between these two species, as well as in sensilla size and number. Based on the morphology of the proboscis and associated sensilla, S. trifolii and P. scutosa are potential flower visitors, which was also reinforced by the pollen observed at the proboscis tip. These results will strengthen our understanding of the structure of the proboscis related to the feeding behavior of Noctuidae.The proboscis is an important feeding organ for the glossatan moths, mainly adapted to the flower and non-flower visiting habits. The clover cutworm,  Insect mouthparts are modified appendages of head segments, bearing various types of sensory organs and adapted to exploit different food resources . Due to In adult Glossata, the proboscis has a relatively simple morphology, but it is unique in its coiled resting position. The proboscis consists of two extended concave maxillary galeae and a hollow food tube, which are joined by dorsal and ventral cuticular projections, i.e., legulae ,14. The sensu stricto) is the second largest family in Lepidoptera, including numerous agricultural pests of great economic significance [Scotogramma trifolii Rottemberg, and the spotted clover moth, Protoschinia scutosa (Denis & Schiffermuller), are worldwide polyphagous pests in the subfamily Noctuinae and Heliothinae, respectively [S. trifolii and P. scutosa have characteristics of intermittent local outbreaks and migration [S. trifolii and P. scutosa feed mainly on many agricultural crops, such as Gossypium herbaceum Linnaeus [Chenopodium quinoa Willd [S. trifolii and P. scutosa, but it has never been satisfactorily explored.Noctuidae . The proboscis was cut from the head with dissecting scissors under a stereomicroscope . The samples were fixed in Carnoy’s fixative solution  for 24 h, and then immersed in 75% ethanol solution.Adults of The proboscis was washed using an ultrasonic cleaner  for a few seconds. All samples were dehydrated through a graded ethanol series of 80%, 85%, 90%, and 95% for 10–15 min each, and 100% for 30 min, twice. The proboscis was naturally dried on filter paper. The dried samples were mounted at various angles using double-sided graphite adhesive tape, and sputter coated with gold. All samples were examined under a Hitachi S-3400N scanning electron microscope  at 5.00 kV and full vacuum.t-test was used to evaluate differences of proboscis and associate sensilla between species and sexes (p < 0.05) in SPSS Statistics v 22.0.0.0. The software Photoshop CS  was used to adjust contrast and levels of SEM images.The proboscis dimensions and sensilla size were measured using Imaris 7.4.2 software with seven males and seven females of each species. An independent sample Morphological descriptions of the proboscis follow the terminology of Lehnert et al.  and FaucS. trifolii and P. scutosa are nearly same in shape and structure, both consisting of two elongated and coiled maxillary galeae (P. scutosa (S. trifolii and P. scutosa (p > 0.05)  .S. trifolii and P. scutosa , which are present on both  scutosa . Zone 1  scutosa A–C. Zone scutosa D,E. The  scutosa C. S. trisensilla . No signsensilla . P. scuttrifolii .S. trifolii and P. scutosa . The proboscis structure is relatively conservative in Noctuidae, and sensilla morphology may not be useful for comparative studies of nearby species. In Noctuinae, Mythimna separata [Athetis lepigone [S. trifolii possesses Zone 3 , and possesses three types of sensilla, sensilla chaetica, basiconica, and styloconica. However, S. trifolii and P. scutosa exhibit significant differences in the length of the proboscis and each zone, as well as in the dimension of each sensillum type. The presence of floral pollen, in combination with the characteristics of the distal proboscis indicates that S. trifolii and P. scutosa are very likely flower visitors. These findings provide a morphological basis to better understand the feeding mechanisms of both species.This study shows that the proboscises of


Ixodes ricinus ticks in Wester Ross, Northwest Scotland, a region of high tourism and tick exposure, yet data-poor in terms of tick-borne pathogens.Lyme borreliosis and other tick-borne diseases emerge from increased interactions between humans, other animals, and infected ticks. The risk of acquiring a tick-borne infection varies across space and time, so knowledge of the occurrence and prevalence of pathogens in ticks can facilitate disease diagnosis in a specific area and the implementation of mitigation measures and awareness campaigns. Here we identify the occurrence and prevalence of several pathogens in I. ricinus nymphs (n = 2828) were collected from 26 sites in 2018 and 2019 and tested for the presence of tick-borne pathogens using PCR-based methods. Prevalence was compared with other regions of Scotland, England, Wales, and the Netherlands.Questing Anaplasma phagocytophilum , Borrelia burgdorferi sensu lato (s.l.) (2.2%), Babesia from clade X (0.2%), Rickettsia helvetica (0.04%), and Spiroplasma ixodetis (0.4%) were detected, but no Neoehrlichia mikurensis, Borrelia miyamotoi, or Babesia microti. Typing of A. phagocytophilum using a fragment of the GroEL gene identified the presence of both ecotype I and ecotype II. Genospecies identification of Borrelia burgdorferi s.l. revealed B. afzelii (53% of infected nymphs), B. garinii (9%), B. burgdorferi sensu stricto (7%), and B. valaisiana (31%). We found similar prevalence of A. phagocytophilum in Wester Ross as in the Netherlands, but higher than in other parts of Great Britain. We found lower B. burgdorferi s.l. prevalence than in England or the Netherlands, and similar to some other Scottish studies. We found higher prevalence of B. valaisiana and lower prevalence of B. garinii than in other Scottish studies. We found S. ixodetis at much lower prevalence than in the Netherlands, and R. helvetica at much lower prevalence than in England and the Netherlands.S. ixodetis in Great Britain. The results are relevant for disease surveillance and management for public and veterinary health. The findings can also aid in designing targeted public health campaigns and in raising awareness among outdoor recreationists and professionals.As far as we know, this is the first description of The online version contains supplementary material available at 10.1186/s13071-021-04946-5. Ixodes ricinus is the most abundant and widespread tick species in Europe [Borrelia burgdorferi sensu lato (s.l.) infection, and tick-borne encephalitis (TBE), both of which have increased in incidence in several European countries in recent decades [n Europe  and, as n Europe . The pri decades –7. TBE v decades .Ixodes ricinus also transmits other pathogens, including Anaplasma phagocytophilum, Borrelia miyamotoi, Neoehrlichia mikurensis, Rickettsia helvetica, Spiroplasma ixodetis, and several Babesia species. These are currently the key known pathogens that can cause disease in livestock, wild animals, and/or humans [r humans , 10. Altr humans .B. miyamotoi and TBE virus in questing ticks in the Netherlands, local health professionals identified the first cases of hard tick-borne relapsing fever and TBE in human patients and could thus administer appropriate treatment [Information on the geographical distribution and prevalence of pathogens in ticks helps to evaluate the risk of exposure through tick bites and, consequently, the risk of disease. This raises awareness of the appropriate tick-borne pathogens in areas of risk and can contribute to prevention, identification, diagnosis, and treatment of these diseases in humans and animals. This is particularly relevant to regions where the pathogens, and hence the accompanying diseases, are currently considered absent due to the lack of data on the presence and prevalence of these pathogens in ticks. For example, soon after becoming aware of the presence of reatment , 13. AwaOne region in Europe that is data-deficient in terms of these emerging pathogens is Northwest Scotland. It is an area of potential high relevance to tick-borne pathogens, as it has high tick host populations, both wild, such as deer and small mammals, and livestock, especially sheep . NorthweI. ricinus ticks from Wester Ross, including a preliminary estimate of their prevalence. Specifically, we aim to test I. ricinus for the presence of A. phagocytophilum, B. burgdorferi s.l., B. miyamotoi, N. mikurensis, R. helvetica, S. ixodetis, Ba. microti, and Babesia species from clade X, formerly known as Babesia sensu stricto (s.s.) [A. phagocytophilum, B. burgdorferi s.l., and Babesia species from clade X are further specified to the ecotype, genospecies, and species level, respectively. Importantly, we place these new presence and prevalence data in context by comparing them with previously published and other unpublished data from other regions of Scotland, England and Wales, and the Netherlands, as an example from continental Europe.The aim of this study is to provide a first assessment of the presence of a range of pathogens in questing o (s.s.) –19. SampPinus sylvestris) woodlands (15–20 years old), mature Scots pine woodlands (minimum 50 years old), mature birch (Betula spp.) woodlands, and open moorland sites dominated by heather . A total of 2828 questing I. ricinus ticks  were collected using the blanket drag/cloth lure method and kept for analysis in Eppendorf vials containing 70% ethanol. Previous studies have confirmed that 100% of the ticks identified from blanket drags in Scotland, including the Northwest region, are I. ricinus: in woodland sites across Scotland including Northwest Scotland, 2000 questing nymphs identified to species level were I. ricinus [I. ricinus [I. ricinus [I. ricinus. All the 2828 questing ticks were used for molecular analysis. For detailed information on the geographical location, land cover type, and times of data collection on the sites, see Fig. Ticks were collected from 26 sites in Wester Ross in Northwest Scotland, United Kingdom , in 2018 and 2019  were then washed in distilled water, briefly dried, separated individually into polymerase chain reaction (PCR) plates, and kept at –20 °C until DNA extraction. All ticks were extracted with ammonium hydroxide as described previously [B. burgdorferi s.l. [B. miyamotoi [N. mikurensis [A. phagocytophilum [Babesia microti [S. ixodetis [R. helvetica [Babesia spp. from clade X, which has been designed to detect Ba. divergens, Ba. venatorum , Ba. capreoli and Ba. odocoilei [B. burgdorferi s.l. were subjected to conventional PCR and Sanger sequencing of the intergenic spacer region for genospecies identification [A. phagocytophilum were subjected to conventional PCR and Sanger sequencing of a fragment of the GroEL region for ecotyping [Ticks were kept in Eppendorf vials filled with 70% ethanol until return from the field. All ticks . For additional B. burgdorferi prevalence in Scotland and for the prevalence of all the pathogens in England and Wales, data from published studies were used [Anaplasma phagocytophilum and B. burgdorferi s.l. nucleic acids were subjected to specific TaqMan RT-PCR [A. phagocytophilum [B. burgdorferi s.l. in our Wester Ross ticks [Babesia spp. DNA was amplified according to Hilpershauser et al. (2020) [https://blast.ncbi.nlm.nih.gov/Blast.cgi) to obtain parasite speciation. Prevalence estimates of nymphs assumed that a positive pool resulted from one positive nymph, i.e. the number of positive pools divided by the number of nymphs tested for each site. The overall prevalence for each pathogen cited in the results from the Gilbert and Rocchi data was the mean of all the site-level prevalence values.We sourced data from different studies to compare the presence and prevalence of the tested pathogens in questing herlands  as an exere used , 34–43. ere used , 40–43 an RT-PCR , which itophilum  but diffss ticks . Babesia. (2020) . PositivI. ricinus ticks from Wester Ross, from highest to lowest prevalence: A. phagocytophilum , B. burgdorferi s.l. , S. ixodetis , Babesia spp. from clade X , and R. helvetica . See Additional file Anaplasma-positive ticks, 86% (80/93) were identified as the zoonotic ecotype I and 14% (13/93) as the non-zoonotic ecotype II. Four genospecies of B. burgdorferi s.l. were detected in the following proportions of positive nymphs: B. afzelii , B. valaisiana , B. garinii , and B. burgdorferi s.s. . Two genospecies of Babesia from clade X were detected, in the following proportions of positive nymphs: Ba. venatorum  and Ba. divergens .We detected the following pathogens tested in the 2828 questing The presence and prevalence of the tick-borne pathogens from other regions of Scotland, England and Wales, and the Netherlands are summarised in Table Anaplasma phagocytophilum was present at a higher prevalence than in studies from England, and at a similar prevalence as the Netherlands. We found B. burgdorferi s.l. prevalence to be lower than in England or the Netherlands, and similar to some other Scottish studies. We found a higher prevalence of B. valaisiana and lower prevalence of B. garinii than in other studies from Scotland (not shown in table). We found S. ixodetis at much lower prevalence than in the Netherlands, and R. helvetica at much lower prevalence than in England and the Netherlands.A. phagocytophilum, B. burgdorferi s.l., Babesia species from clade X (Ba. venatorum and Ba. divergens), R. helvetica, and S. ixodetis, the last being the first record described for Great Britain. This study assessed only the presence of the DNA from the chosen pathogens, and not their viability or infectivity. However, numerous studies already implicate I. ricinus as their vector. We cannot assume infectiousness from PCR results, and therefore we cannot translate prevalence estimates of the pathogens to infection risk in humans.We revealed the presence of the following tick-borne pathogens in ticks in Wester Ross, Northwest Scotland: A. phagocytophilum, with a prevalence higher than in England and Wales, but comparable to locations in the Netherlands and elsewhere in Europe [I. ricinus [Capreolus capreolus), which are present at our survey sites in Wester Ross [Anaplasma phagocytophilum ecotype I is probably zoonotic and can also cause disease in livestock. It is probably maintained in enzootic cycles by deer species other than roe deer [Cervus elaphus) are the most common deer species, and likely to be the main natural reservoir host.The most frequently detected pathogen group was n Europe . The two ricinus , and we ter Ross . Anaplasroe deer . In NortB. burgdorferi s.l., with a prevalence of 2.2%. This prevalence is comparable to that reported in two of the four other large cross-sectional surveys from further south and east in Scotland, in the Grampian region (2.2%) [The second most abundant group of tick-borne pathogens was n (2.2%) , and acrn (2.2%) . The othn (2.2%) , and 5.9n (2.2%) . This hin (2.2%) .B. afzelii was the genospecies that constituted most (53%) of the B. burgdorferi s.l. complex of spirochetes in Wester Ross.Similarly to other studies in Scotland , 34, 43,B. burgdorferi s.l. complex was comprised of B. burgdorferi s.s., which is a similar proportion as that reported in all the large Scottish surveys [B. burgdorferi s.s. zoonotic genospecies in Europe has not been firmly established, but recent studies, including from Scotland, suggest a dominant role for squirrels [We found that in Wester Ross, 7% of the  surveys , 34, 43.quirrels –50.B. garinii and B. valaisiana, comprised markedly different proportions in Wester Ross from those in previous Scottish surveys, which showed far greater proportions of B. garinii  are similar to those in the other Scottish surveys, and are expected to comprise similar bird species, so we do not know why B. valaisiana dominates in Wester Ross as compared to the apparent dominance of B. garinii in the rest of Scotland. One possible explanation is competition between the pathogens within the host or the vector, and/or that there is a certain amount of host specificity of each of the bird genospecies between bird species. There is some evidence that B. garinii can associate with squirrels [Interestingly, the two bird-associated genospecies, i  and smana . We fouquirrels , so perhquirrels .Borrelia afzelii is associated with a skin condition, acrodermatitis chronica atrophicans (ACA), B. garinii with neuroborreliosis, and B. burgdorferi s.s. with Lyme arthritis [B. valaisiana may not be pathogenic [B. afzelii, general practitioners need to be aware of ACA and its manifestations.The genospecies composition is relevant from a public health perspective, as different genospecies lead to different clinical symptoms in human Lyme disease patients. rthritis , 52, 53,thogenic . As we fB. burgdorferi s.l.-positive ticks in this study were from mature birch and mature Scots pine woodlands, and both these habitat types are likely to support diverse vertebrate communities, but may differ in their species composition. The low bird-associated B. garinii prevalence suggests that rodents play a more important role in Lyme disease hazard than do birds in the study area. Further research on bird and rodent densities at the sites could elucidate the mechanisms driving the relative composition of Borrelia burgdorferi s.l. genospecies in Wester Ross.The genospecies composition is also interesting from an ecological perspective. Habitat is an important determinant of vertebrate community composition. All the Babesia species from clade X were detected in Wester Ross at a prevalence of 0.2%, compared to 0.6% in other regions of Scotland . Two of the four common Babesia species from I. ricinus were detected. The first, Ba. venatorum, has been associated with some disease cases in humans in Europe [, although the main natural vertebrate host for Ba. venatorum is probably roe deer in Europe [Babesia divergens, the second detected genospecies, is of zoonotic importance: it has several zoonotic reservoirs, including deer and cattle, both of which have high cultural and economic value in Wester Ross. It has been found all across Europe affecting cattle [Ba. divergens in the UK was reported in Southwest England [Babesia capreoli and Ba. odocoilei were not detected, but they are present in the Netherlands. No human cases have been associated with either of the two latter pathogens in Europe [n Europe , 56 and n Europe  and dogsn Europe  in the Un Europe , 60. Babg cattle , 62 as wg cattle , 63–65,  England . Babesian Europe .R. helvetica in Wester Ross, Northwest Scotland. This pathogen has previously been detected in ticks in England at a 3% prevalence [R. helvetica is efficiently transmitted vertically in ticks , ticks themselves can be considered as reservoirs hosts [R. helvetica has not be fully investigated yet, but the pathogen has been detected in bank vole, wood mouse, shrew, wild boar, and roe deer [R. helvetica is very low, and one may wonder whether or how a tick-borne pathogen can be maintained in an enzootic cycle with such a low prevalence. One possible explanation is that ticks with R. helvetica are not maintained in a regional enzootic cycle but (continuously) introduced by migratory birds [We found very low prevalence (0.04%) of evalence  and in sevalence , Germanyevalence , Austriaevalence , Slovenievalence , and Spaevalence , and hasevalence –76. It ievalence . As R. hrs hosts . Which vroe deer , 78, 79.ry birds .S. ixodetis , and this is the first record for Great Britain. The bacterial parasite Spiroplasma ixodetis was first isolated from Ixodes pacificus ticks in the USA in 1995 [I. ricinus including larvae [, which suggests that transmission can be transovarial. The role of hosts in the S. ixodetis transmission cycle is unknown [S. ixodetis [We detected the relatively unknown tick-borne pathogen  in 1995 , and it g larvae –84, whic unknown . Severalixodetis , 86.N. mikurensis, B. miyamotoi, or Ba. microti in questing nymphs from Wester Ross, this cannot necessarily be interpreted as a complete absence of enzootic circulation of these pathogens, because the prevalence of some of these pathogens can be low, for example, 0.1% for N. mikurensis in regions of Denmark [Ba. microti in Poland [B. miyamotoi in regions of Germany [Ba. microti in one engorged tick pulled from a wood mouse (Apodemus sylvaticus) . Neoehrlichia mikurensis has so far not been found in any study that tested for it in Great Britain [Although we did not observe  Denmark , 0.6% fon Poland  and 1.5%n Poland , and 0.8 Germany  and 0.5% Britain , 37, so ricinus, although this is highly unlikely, as previous studies from Scotland identified 100% of questing ticks  to be I. ricinus [.We tested 8282 questing nymph and adult ticks, and it is possible that a tiny proportion of the ticks analysed may have been non- ricinus –22.A. phagocytophilum, B. burgdorferi s.l., Babesia species from clade X (Ba. venatorum and Ba. divergens), R. helvetica, and S. ixodetis, the latter being the first record described for Great Britain. Anaplasma phagocytophilum occurred at higher prevalence than elsewhere in Great Britain, while B. burgdorferi s.l. was less prevalent than in England or the Netherlands, though similar to other Scottish studies. This new knowledge has implications for public and veterinary health in terms of improving diagnoses and treatment and better targeting of awareness and mitigation strategies.In the first surveys of tick-borne pathogens in Northwest Scotland, in a region much frequented by tourists, we identified and quantified the prevalence of Anaplasma phagocytophilum and Ba. divergens, both of zoonotic importance, are transmitted by deer, an important cultural and economic asset in the Wester Ross area as well as other parts of Europe. Rickettsia helvetica has been found in several vertebrates, but its life cycle is unclear and needs further investigation. More studies in different habitats are needed to shed light on the ecological determinants of genospecies composition of B. burgdorferi s.l.Further research is needed to investigate the life cycles and mechanisms of transmission of these pathogens. Additional file 1:Text S1. Field methods explained. Table S1. Sum of ticks collected at each visit on each plot is summarised, as well as the sum of ticks positive for each of the eight pathogens tested for in the study, namely Borrelia miyamotoi, Borrelia burgdorferi s.l., Anaplasma phagocytophilum, Neoehrlichia mikurensis, Babesia spp. from clade X, Spiroplasma ixodetis, Babesia microti, and Rickettsia helvetica. Table S2. Molecular analysis results of the 2828 adult and nymph ticks.


Tetratrichomonas gallinarum is parasitic protozoa with a wide host range. However, its lethal infection is rare reported.T. gallinarum infection in black swans in China. Five black swans died within a week in succession without obvious symptoms except mild diarrhea. At necropsy, severe lesions were observed in caeca with thickened caecal walls and hemorrhages in the mucosa. A large number of moving trophozoites were found in the contents of the cecum by microscopic examination. The livers were enlarged with multiple bleeding spots on the surface. Histopathology of the livers showed mononuclear cell infiltration and moderate hyperplasia of fibrous tissue. The histopathology of the cecum showed that the villi of the cecum were edematous. Finally, the presence of T. gallinarum was determined by specific PCR andin-situ hybridization assay. Additionally, common pathogens that can cause similar symptoms were excluded.Here, we described the first lethal cases of T. gallinarum, suggesting that the parasite might be a new threat to the Cygnus birds.The death of the black swan was caused by  Tetratrichomonas gallinarum is parasitic protozoa with a wide host range [T. gallinarum is controversial [T. gallinarum in birds were sporadically reported in some countries, such as in chukar partridges, mockingbird, Waldrapp ibis and white pelican from America [Cygnus atratus) associated with T. gallinarum infection in China, and the threat of the protozoa to Cygnus birds must be considered.st range . Owing toversial –4. Moreo America –8, in du America , in red- America , and in  America . Here, wIn August 2019, five adult black swans from a wetland park of Beijing died within a week in succession. Before they died, no obvious symptoms were observed except mild diarrhea.The fresh carcasses were sent to the National Research Center for Wildlife Borne Diseases for postmortem and histopathological examination. At routinely pathological investigation, the ceca were swollen and the mucosa were hemorrhages and anabrosis Fig. a. A largT. gallinarum and H. meleagridis [T. gallinarum probe were found in the caeca  using the described probe specific for eagridis , 13. Theeca Fig. c but noteca Fig. d. The reT. gallinarum download from GenBank database under phylogenetic analyses  [T. gallinarum recovered from the black swans may be spillover from the ducks.Studies conducted by Dimasuay and Rivera shown that hynchos) , which sT. gallinarum infection in China, suggesting that the protozoan might be a new threat to the Cygnus birds. A comprehensive epidemiological investigation of T. gallinarum in Cygnus birds is urgently needed in the future.In conclusion, we described the first fatal case of black swans associated with


Deliaplanipalpis (Meigen) and D.platura (Stein). They are a threat to the production of these vegetables since the damage they cause directly and indirectly affects yield, quality, and commercialization of these crops. Nevertheless, the existence of natural enemies of these dipterans is still unknown. It is fundamental to determine which parasitoids or predators can be considered possible biological control agents in an integrated pest management program.Among the insect pests that affect crucifer crops in Mexico are BrassicaoleraceaL.var.italica Plenk and capitata L., B.napus L., and Raphanussativus L. infested with Delia spp. were selected. The symptoms observed were wilting, yellowish, flaccid leaves and individuals less developed than the rest of the crop. These plants were extracted with their root and the surrounding soil. Also, wild crucifers were collected, such as Raphanusraphanistrum L., Brassicacampestris L., and Sisymbriumirio L. The first records of Aphaeretapallipes Say , Trybliographarapae (Westwood) , and Aleocharabimaculata Gravenhorst  are reported parasitizing the puparia of these anthomyiid flies on cultivated and wild crucifers. This represents only a starting point for the continuous study of these parasitoids, which is needed to consider them useful for the biological control of D.planipalpis and D.platura.The sampling sites were located in Guanajuato, Puebla, and the State of Mexico, where plants of  Delia (Robineau-Desvoidy) , commonly called root maggots, attack economically important crops of the family Brassicaceae , North America (Canada and the United States), Asia , and Africa (Algeria and Morocco) , sister species of D.radicum and D.floralis  (Raphanussativus L.) , cabbage (B.oleraceavar.capitata L.), cauliflower (B.oleraceavar.botrytis L.), radish, turnip (B.napus L.), and wild crucifers such as field mustard (B.campestris L.), wild radish (R.raphanistrum L.), and London rocket (Sisymbriumirio L.) ) (D.florilega (Zetterdest) and D.platura (Stein), which have a wider range of hosts and can feed on decomposing tissue. However, under certain circumstances, they also feed on healthy tissues of plants of the family Brassicaceae and species of the genus Allium L., as well as legumes, Cucurbitaceae, and cereals (Some species of the genus sicaceae . These isicaceae . One exaMorocco) . It is oMorocco) . Another , whose mivus L.) . Howeveririo L.) . NeverthMeigen)) . There a cereals .D.planipalpis and D.platura associated with crucifers in Mexico. Larvae of these dipterans cause damage mainly to the root crown of some cultivars of B.oleracea, and consequently, they affect the root system. Occasionally, the larvae develop near the broccoli head causing rot and malformation, or no formation of the inflorescence. In addition, in crops such as radish and turnip, direct damage caused by larvae to their edible part makes their commercialization difficult ) is grown in Canada , cabbage (B.oleraceavar.capitata L.), turnip (B.napus L.), radish (Raphanussativus L.), and other wild crucifers such as field mustard (B.campestris L.), London rocket (Sisymbriumirio L.), and wild radish (R.raphanistrum L.).The study was conducted between February 2018 and February 2019 in Guanajuato, Puebla, and the State of Mexico, where some sites infested by Delia spp. Wild crucifer plants were selected at random within and on the outer edges of commercial crops and the sample size varied from 5 to 10 plants depending on their abundance. Each plant was extracted with its root and adhered soil and placed in a polyethylene bag; additionally, with a post hole digger of 15.0 cm in diameter by 20.0 cm long, the soil of the first 10.0 cm of depth was collected taking as a reference point the exact site where the plant was extracted and because in this place the puparia are distributed due to the limited movement of third instar larvae around the plant and from the soil surface , Dr Fabiana Gallardo , and Dr José Luis Navarrete Heredia . The material is kept as reference specimens in the entomological collection of the Colegio de Postgraduados Campus Montecillo.The keys of Deliaplanipalpis and 49 D.platura puparia were collected in cultivated and wild crucifers in the states of Guanajuato, Puebla, and the State of Mexico , Trybliographarapae (Westwood) , and Aleocharabimaculata Gravenhorst    , and theA.verna  are nameA.pallipes has not been mentioned as an important parasitoid of Delia spp. in Europe and Canada, A.pallipes and A.bilineata caused mortalities of up to 17 and 20%, respectively, in D.antiqua in commercial onion crops in southwestern Ontario. This braconid has a wide range of hosts and prefers members of at least three families, Coelophidae, Calliphoridae and Anthomyiidae . In addition, it has been reported that this staphylinid also attacks Adiacinerella Fallen  and Scatophagastercolaria L.    , as welluscidae) . Even thuscidae) . A.bimaribution  and thatribution , it is e develop .B.campestris and S.irio, alternate hosts are important reservoirs of considerable populations of Delia spp. and consequently of their natural enemies, especially because they provide habitat during the season when there are no crops. In this way, wild crucifers assure that there is no scarcity of hosts for either the insect pest or their parasitoids and predators (Finally, although in this work no parasitoids were found in the wild crucifers D.planipalpis and D.platura collected in cultivated and wild crucifers were identified: one gregarious parasitoid, Aphaeretapallipes and two solitary parasitoids, Trybliographarapae and Aleocharabimaculata. Only Aphaeretapallipes and T.rapae were specific to D.planipalpis and D.platura, respectively. Finally, A.bimaculata was associated with both pest species.Three species that emerged from puparia of D.planipalpis and D.platura that occur naturally in Mexico. Aphaeretapallipes was the most abundant species, found in Guanajuato and Puebla, followed by Aleocharabimaculata, and finally T.rapae, which was found only in the State of Mexico. Knowledge of natural enemies of Delia spp. in crucifer-producing regions is only the starting point from which to broaden the search and begin to study their biological attributes with the aim of incorporating them into a biological control program against D.planipalpis and D.platura.This is the first record of parasitoids of


Introduction:Rhabdophis snakes, which include 27 species, are rear-fanged venomous snakes that are widely distributed from India to East Asia and Russia. Severe envenomation by R. tigrinus (Yamakagashi snake) in Japan and R. subminiatus in Southeast Asia has been reported. The epidemiology of R. tigrinus bites, such as geographical features, the incidence, and changes in the number of bites over time have not been comprehensively examined. Hence, we intended to clarify the epidemiological features of R. tigrinus bites through a careful review of scientific data over the last 50 years in Japan.Methods: Patient records of R. tigrinus bites between 1971 and 2020 at the Japan Snake Institute were examined retrospectively. The following were ascertained: patient characteristics, clinical symptoms, laboratory data, treatment-related factors, and hospital mortality. These variables were compared in the antivenom and the without-antivenom groups.Results: Over the 50-year study period, 43 R. tigrinus bites, including five fatal cases, were encountered. Severe cases of R. tigrinus bites have been treated with antivenom since 1985; however, fatalities occurred in 2006 and 2020. R. tigrinus bite cases have been well-distributed in the western part of Japan since 2000. The mortality rate in the antivenom group was significantly lower in the patient group that was not administered the antivenom .Conclusion: This study clarified the epidemiology of R. tigrinus bites in Japan over a 50-year period. Almost all severe cases of R. tigrinus bites have been treated with the antivenom in the current situation, and fatalities occurred in cases not treated with the antivenom. It is important to diagnose R. tigrinus bites in the early phase of the clinical course. The antivenom, the definitive treatment for R. tigrinus bites, is an unapproved drug. Hence, approval needs to be obtained for the drug. Rhabdophis include 27 species of rear-fanged venomous snakes that are widely distributed from India to East Asia and Russia. Severe envenomation by R. tigrinus (Yamakagashi snake) in Japan and R. subminiatus in Southeast Asia have been reported  has also been evaluated from 1973 to 2013  was founded in the Gunma Prefecture to conduct research and provide enlightenment on snakes . This inR. tigrinus bites was based on the patients' description of the snakes and the presence of VICC in patients with hypofibrinogenaemia.The diagnosis of Data on age, sex, date of injury (year), place of injury, clinical symptoms, laboratory analyses, treatment-related factors, and hospital mortality were recorded.U-test or the Chi square (χ2) test was used. Furthermore, the Fisher exact test for 2 × 2 categorical variables was used when appropriate. P-values < 0.05 were considered statistically significant. All analyses were completed using the JMP version 11  software.To compare the data pertaining to patient characteristics, treatment-related factors and outcomes between the antivenom patient group and the without-antivenom group, the Mann–Whitney R. tigrinus bite patients were recognized. The median age was 37 years, and 97.7% were men.R. tigrinus bite cases were found in almost all parts of Japan, except Hokkaido and Okinawa areas, and have been distributed in the western part of Japan since 2000. Fatalities have been limited to the western part, and specifically, there have been two fatalities in the Okayama Prefecture .R. tigrinus antivenom. Cerebral hemorrhage was observed in least three out of five cases.Details of fatalities are provided in R. tigrinus bites, including five fatalities, were identified. Patients with R. tigrinus bites have been treated with the antivenom since 1985, and fatalities occurred in 2006 and 2020 in patients who were not treated with the antivenom.Over the 50-year study period, 43 cases of R. tigrinus bite cases have been well-distributed in the western part of Japan since 2000. Fatalities have also been limited to the western part. This finding is similar to the cases of pit viper bites reported previously  20fk0108101.KM was employed by company KM Biologics Co., Ltd. The remaining 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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Neurosurgical Review,Dear highly esteemed readership and editorial team of Neurosurgical Review in 2016, the journal went through a number of difficult, but necessary, changes. After the journal’s remarkable success story under the editorship of Helmut Bertalanffy, who had worked to achieve this success, Springer Nature, and more precisely Springer-Verlag GmbH, changed the format of the journal from print to exclusively online. By also changing the managing editorial software, the editorial team introduced a number of new and important features, such as a plagiarism check for any manuscript submitted since 2017. In the beginning, my personal aim was to further promote the already high quality of the journal and guide it to becoming the journal with the highest impact factor among all the European neurosurgical journals.When I took over responsibility as Editor-in-Chief of Neurosurgical Review: Editorial Office, Sandra Braun; and Editorial Assistants , PD Dr. Y. Ahmadipour, Dr. L. Barthel, Dr. B Chen, PD Dr. M. Darkwah Oppong, Dr. O. Gembruch, Dr. S. Hetze, PD Dr. R. Jabbarli, PD. Dr. D. Pierscianek, and Dr. L. Rauschenbach. I am most grateful to all these people for their commitment and excellent work. In addition, Springer Verlag GmbH has supported my efforts to improve the quality of Neurosurgical Review by providing a highly qualified and motivated managing team at the publisher end; I would particularly like to highlight the merits of Dr. M. F. Bartels.To do so, I needed considerable support from local collaborators, and I therefore built up a highly motivated editorial team in Essen. In the past 5 years, this editorial team consisted of numerous important “helping hands and minds.” Most recently, the following colleagues have contributed to the success of Neurosurgical Review succeeded in dramatically increasing the number of submitted manuscripts (nearly 4-fold), the number of published Online First articles (2-fold), and the article downloads (2-fold)  to 3,042: an all-time high in the history of My mission is accomplished!So, in summary, after a demanding and strenuous 5 years as editor, I would like to declare:Neurosurgical Review, has an even greater potential for further development. I hope that our readership, as well as all our contributors, agrees with me when I say:Yes, the mission is accomplished, but not yet finished!However, I believe that our originally European, and increasingly international journal, Thank you all once again for your constant, and hopefully continued, support!Ulrich Sure,Essen, Germany


Mycotoxin-Workshop 2021 was held on 31st May–2nd June, as an online conference. Unfortunately, last year’s mycotoxin workshop, which was planned in Brno (Czech Republic) had to be cancelled due to the COVID-19 pandemic. This year, an online forum for the presentation of new developments in mycotoxin research, as well as for the discussion of future perspectives in this field of research, was provided by the team of Prof. Dr. Hans-Ulrich Humpf, who is affiliated with the Institute of Food Chemistry, University of Münster. As always, the meeting was organised in cooperation with the Society for Mycotoxin Research. The scientific programme was scheduled for 3 days and contained nine main half-day sessions and six parallel short-talk poster sessions, covering contemporary mycotoxin research topics. This year, the Mycotoxin Workshop attracted the attention of almost 300 virtual participants from 31 countries worldwide, which resulted in 40 lectures and 69 short poster-style presentations. The online format of the Mycotoxin Workshop enlarged the geography of the members giving the opportunity to share the research with colleagues not only from EU, but also from Brazil, Canada, Chile, China, Egypt, Kenya, Saudi Arabia, South Africa, and the USA .The award committee honoured the doctoral thesis of Ms. Jagels “Isolation and structure elucidation of new secondary metabolites from Society for Mycotoxin Research. The award is open to scientists worldwide, but the application of the candidate must be supported by a member of the Society for Mycotoxin Research. All details for the next call will be provided on the home page of the Society for Mycotoxin Research (www.mycotoxin.de).This highly prestigious prize in the field of mycotoxin research is endowed with 10,000 Euro and given biennially by the Society was held during the 42nd Mycotoxin Workshop. The board of directors reported about the number of members and the financial situation of the Society. Currently, the Society has about 180 members from 26 countries with increasing tendency especially from young scientists. The Society for Mycotoxin Research is a non-profit organisation focusing on scientific excellence and exchange in the field of mycotoxin research and supporting particularly young scientist. Members benefit from a free print and online subscription of the official journal of the Society, Mycotoxin Research, as well as a highly reduced registration fee for the Mycotoxin Workshop. Prof. Dr. Ewald Usleber, the Editor in Chief of Mycotoxin Research, gave a short report about the current situation with the official journal of the Society. Mycotoxin Research and its impact for the scientific community continue to develop very well. For example, the impact factor  increased from 3.1 in 2019 to 3.8 in 2020.The annual meeting of the Mycotoxin Research, Prof. Dr. Ewald Usleber was elected as honorary member of the Society.For his excellent job as Editor in Chief of Society, the Board of Directors was newly elected. The current President of the Society, Prof. Dr. Dr. habil. Manfred Gareis, who served the Society for 20 years since its foundation in 1997, wished to step back from his position. Prof. Dr. Hans-Ulrich Humpf expressed his willingness to follow Manfred Gareis as President. In a unanimous vote, the following Board of Directors was (re)-elected for the period 2021–2023:According to the bylaws of the President: Prof. Dr. Hans-Ulrich Humpf, Münster, Germany.Vice President: Prof. Dr. Madeleine Plötz, Hannover, Germany.Secretary: Dr. Karsten Meyer, Freising, Germany.Treasurer: Dr. Benedikt Cramer, Münster, Germany.Society for Mycotoxin Research.Prof. Humpf thanked Prof. Gareis for his outstanding and long-lasting support of the Mycotoxin Workshop will take place on May 30–June 1, 2022, in Toulouse, France. The conference will be organized by the groups of Dr. Isabelle Oswald, Prof. Florence Mathieu and Dr. Mohamed Haddad in cooperation with the Society for Mycotoxin Research. For further information concerning registration, abstract submission, deadlines, and fees, please check the website of the Society for Mycotoxin Research (www.mycotoxin.de).The 43rd


Neospora caninum is one of the most common infectious organisms worldwide that causes abortion in cattle. To the best of our knowledge, no previous studies have focused on N. caninum infection in the local Atlas brown cattle from Northeast Algeria. This study aimed to assess the importance of bovine neosporosis for causing abortion in Atlas brown cattle and to identify selected risk factors.N. caninum using enzyme-linked immunosorbent assays.A case-control study was performed on 60 control farms and 30 case farms. We collected 650 blood samples from 650 pregnant cows from 90 farms in five Algerian provinces; Jijel, Skikda, Annaba, El-Tarf, and Souk-Ahras. Sera samples were analyzed for the presence of antibodies against N. caninum infection in the cows was 36.2%  and in the farms was 81.1% (95% CI: 73.0-89.2). Risk factors found by multivariable logistic regression included: Presence of dogs ; age ≥84 months ; Jijel region ; white  and gray  coat; moderate  and bad  hygiene; and second ; and last  stage of pregnancy. Our case-control study showed no significant association between seropositivity of N. caninum and abortion at the farms level . Similarly, there was no significant association between seropositivity of N. caninum and abortion at the individual level .The seroprevalence of N. caninum infection in pregnant local cows from Northeast Algeria. The prevalence rate of antibodies against N. caninum was high. Almost all risk factors studied for infection were significantly associated with seroprevalence. Our analysis showed no relation between N. caninum infection and abortion. Consequently, these local cows are resistant to abortion caused by N. caninum.This is the first study of  Seropositivity to N. caninum was high in these cows. Risk factors for the occurrence of antibodies to N. caninum included dog, age, location, color, and hygiene. Horizontal transmission of N. caninum is probably the most important mode of infection in this region. Our analysis showed no relation between N. caninum infection and abortion; thus, neosporosis should not be used as a differential diagnosis for abortion in these cattle. We recommend increasing the geographical area of study to confirm the conclusions obtained in this study. Research on the mechanism of resistance of Atlas brown cows to abortions will have to be done by genetic study. The future study should be based on identifying the gene(s) involved and the underlying mechanisms.This is the first study of FG, ST: Planned the study. BA: Collected samples, provided serological analyses, analyzed the data, and wrote the manuscript. SYD, AT, and HB contributed to serological analyses. FG: Supervised the study and revised the manuscript. All authors read and approved the final manuscript.


Rhombomys opimus Lichtenstein, 1823) is distributed in Central Asia and some parts of the Middle East. It is widely found in central and northeast parts of Iran with two distinct genetic lineages: R. o. sodalis in the northern slopes of the Elburz Mountains and R. o. sargadensis in the southern slopes. This large rodent acts as the main host of natural focal diseases. No study has surveyed the ecological niche of the lineages and how their distribution might be influenced by different climatic variables. To examine the distribution patterns of this murid rodent, we aimed to determine the habitat preferences and effects of environmental variables on the ecological niche. Using a species distribution approach for modeling of regional niche specialization, suitable habitats predicted for R. o. sodalis were mainly located in Golestan province in northern Iran, along the northern slope of Elburz, while R. o. sargadensis, showed great potential distribution along the southern slope of Elburz and around the Kavir Desert and the Lut Desert. Despite the widest potential distribution of R. o. sargadensis from northeast to northwest and through Central Iran, the geographic range of R. o. sodalis was smaller and mostly confined to Golestan province. The results support the presence of the two genetic lineages of Rhombomys in Iran and confirm that there is no significant niche overlap between the two subspecies. Furthermore, it provided several perspectives for future taxonomic studies and prevention hygiene programs for public health.Great gerbil ( Rhombomys opimus Lichtenstein, 1823), which is generally considered to be a monotypic species based on coloration and size [Rhombomys opimus has a social structure, and individuals often burrow close to one another; the family group  lives in one complex burrow system [Great gerbil , succulent plants such as Salsola spp., Climacoptera spp., and Suaeda spp. (Amaranthaceae) [R. o. opimus  , R. o. giganteus  , R. o. nigrescens  , R. o. fumicolor Heptner, 1933 , R. o. sodalis Goodwin, 1939 , and R. o. sargadensis Heptner, 1939  [This large gerbil has a wide distribution range in central and northeastern Iran and occupies sandy or clay deserts, usually in foothills and mountainous areas with scattered shrubby vegetation, especially thaceae) , 7. The thaceae) , 8. Ellethaceae)  listed sthaceae) . An accuRhombomys opimus sodalis is only found at elevations of approximately 600–1000 m in Golestan province on the northern slope of the Elburz Mountains in addition to small patches in North Khorasan (Bojnourd) and Razavi Khorasan (Sarakhs and Dargaz) provinces in northeast of Iran, whereas R. o. sargadensis, which is considered the more widely distributed subspecies, could be found at higher elevations than R. o. sodalis on the southern slope of the Elburz Mountains and around the Kavir Desert in Central Iran [In Iran, ral Iran , 10, 11 R. o. sodalis in a population of R. o. sargadensis in the Shahrood district (Semnan province), near Kavir Desert [R. o. sodalis from Golestan province to the territories of R. o. sargadensis in Semnan province is possible. However, Oshaghi et al. [R. o. sodalis males and R. o. sargadensis females. However, the reciprocal cross of R. o. sodalis females with R. o. sargadensis males resulted in the death of the paired individuals, suggesting the possibility of some prezygotic isolation mechanism. Bakhshi et al. [R. o. sodalis and R. o. sargadensis [These two subspecies are also morphologically distinct from each other with known differences in coloration and size . Due to r Desert , it has i et al.  and Bakhi et al.  attributi et al.  show thai et al.  obtainedgadensis , and congadensis .Great gerbils can damage crops and irrigation canals, destroy the vegetation and cause the die-off of plants over their colonies by biting the main roots. Hence, they are in direct competition with livestock and have been recognized as carriers of several zoonotic diseases  –14. The R. opimus have different habitat suitability and niche specialization throughout the Iranian Plateau and/or in the contact zone, where they are found in sympatry, and also (ii) to understand the role of environmental variables on their regional spatial distributions.With respect to the lack of information on current habitat delimitation and the potential distribution pattern of great gerbil genetic lineages (or subspecies) in Iran, we seek to elucidate their environmental niches through species distribution models. Accordingly, this study aimed to examine (i) whether the genetic lineages of www.worldclim.org) at a spatial resolution of 1 km2 and then processed to model the target genetic lineage distribution. Since multicollinearity among bioclimatic variables may result in misunderstanding the contribution ratio of the most important variables to the model [r≥ 0.75) were ignored for final analysis. The final set contained 13 climatic variables, as shown in To use a machine learning model, 19 bioclimatic variables were selected from the WorldClim database  , VertNet as a publicly accessible database of vertebrate biodiversity data from natural history collections around the world, and published papers and books e.g., , 22, 23. n = 46) . The poiwww.cs.princeton.edu) was used [To model the current geographic distribution range of great gerbil subspecies, the maximum entropy modeling algorithm . To determine the model performance, the calculated value of the area under the curve (AUC) of the receiver operating characteristic (ROC) curve on the training and testing data was considered. The AUC evaluator indicates the power of the model in distinguishing presence from absence records. A calculated value close to 1 indicates the high predictive ability of the model, while a value of 0.5 suggests that the model lacks sufficient power to predict the species distribution range , 31–34. D  and HellD [I  indices.R. o. sargadensis; AUC = 0.92 for R. o. sodalis), so the MAXENT approach seems to perform well for modeling ecological niche segregation. Based on our models, the most current suitable habitats of R. o. sodalis were mainly projected in Golestan province, along the northern slope of the Elburz Mountains, and penetrated further into North Khorasan provinces as well as small patches in the western parts of Iran  and precipitation of the wettest month (Bio13)  . The resR. o. sargadensis were situated in northeastern, northwestern and Central Iran, while the geographic range of R. o. sodalis was rather smaller and confined mostly to Golestan province, northern Iran. Moreover, the results of ecological niche modeling revealed that there is no significant niche overlap between the two subspecies .In addition, most suitable habitats of great gerbils of the genetic lineage R. o. sargadensis and R. o. sodalis, followed by the annual precipitation amount. However, Gholamrezaei et al. [R. opimus. Furthermore, Gao et al. [R. opimus is distributed in the area of elevation between 200 and 600 m with a slope of 0–3 degrees, an average annual temperature from 6 to 10 °C and an annual precipitation of 120–200 mm in Xinjiang, northwest China.As a desert-adapted rodent, the distribution of great gerbil is significantly associated with temperature, precipitation, terrain, vegetation and other ecological environmental factors . The resi et al.  indicateo et al.  showed tR. o. sargadensis was strongly constrained in an isolated habitat patch in the marginal part of the species distribution range; a patch of habitat in the extreme northwest of Iran might contain genetically isolated populations  . These iHabitat suitability for hosts and/or vectors can have effects on the spreading of pathogens and may influence both their abundance and movements , 43. SuiHarsh climatic conditions (arid or semi-arid climates) in some parts of the Kavir Desert act as another structural barrier for great gerbil to expand its distribution throughout Iran. For the great gerbil, the Elburz Mountains in the north and the Zagros Mountains in the west of the country play major barriers against dispersal.Yersinia pestis, and hence, the transmission rates of plague infection may increase.Lizhi et al.  showed tLeishmania major in Iran, which is transmitted by sand flies of the genus Phlebotomus [L. major is known as one of the zoonoses increasing in Iran [Leishmania in population of great gerbils in Badrood district, Esfahan province, central Iran were surveyed. The lowest and highest infection rates were observed in summer and fall, respectively. Gerbils were found to be infected with three species of L. major, L. turanica and L. gerbilli, which transmit in the population of R. opimus in central part of Iran. Leishmania major infection is generally accompanied by L. turanica in infected great gerbils, showing the highest rate in fall.Great gerbil is considered as the most important reservoir host of ebotomus , 17, 19. in Iran  stated t in Iran , seasonaL. turanica in rodents showed coincidence with the distribution patterns of sandflies [Xenopsylla and Nosopsyllus genera [Hyaloma [Ornithonyssus bacoti [Dentostomella translucida is a nematode parasitizing the digestive system of great gerbils [Hymenolepis nana, which is a cyclophyllidean zoonotic enteric parasite, known to be occurred in different rodents, including gerbils, of Golestan and Razavi Khorasan provinces in north of Iran [The distribution of andflies . Deep buandflies , will haandflies . This ros genera , 55, tic[Hyaloma , 56, ands bacoti –59. More gerbils , 61. Kam gerbils  reported of Iran .Contact zones create candidate habitats where future researches could examine the extent of ecological divergence between the known lineages , 65. TheR. o. sargadensis distributed across most areas of Iran, southern Afghanistan and western Pakistan, the sampling area represents more than half of the geographic range of the subspecies, and the resulting model can potentially be of relatively good quality. However, this is not the case for R. o. sodalis. This subspecies is distributed across northeastern Iran, Turkmenistan and northern Afghanistan. Cumulatively, future studies should consider covering the whole geographical distribution range, especially identifying contact zones, population structure, and comprehensive distribution samplings for genetic studies are required to clarify the taxonomic status of this species. Using a more comprehensive dataset for ecological niche modeling and habitat evaluation will increase the precision of the models and estimate the probable future distribution of the species considering the role of climate changes affecting the environmental variables.The present study provided projections of the potential geographic distribution of the two subspecies of great gerbil, with a fundamental role in the epidemiology of zoonoses. For S1 Appendix(XLSX)Click here for additional data file.


Leishmania tropica is a neglected tropical disease characterized by a wide geographical distribution in the Mediterranean basin and is endemic in several of its countries. In addition, the vector Phlebotomus sergenti is abundantly present all around the basin. Its transmission cycle is still subject to debate. In some countries, the presence of an animal reservoir has been confirmed. In Morocco, CL due to L. tropica has risen since the 1980s and has spread widely to become the most abundant form of leishmaniasis in the territory. However, the anthroponotic transmission is so far the only recognized mode, despite recordings of L. tropica infection in animal hosts. In this review article, we assess the situation of CL due to L. tropica in the Mediterranean basin with a focus on Morocco and gather knowledge about any potential zoonotic transmission in the country. A concomitant zoonotic transmission could explain the persistence of the disease in areas where human protective measures combined with vector management did not help reduce the disease burden.Cutaneous leishmaniasis (CL) due to  Leishmania tropica is a neglected tropical disease still considered to be anthroponotic in Morocco despite evidence of zoonotic transmission in other endemic countries of the Mediterranean basin. Its high incidence in the country makes it necessary to investigate whether or not a potential concomitant zoonotic transmission exists alongside the anthroponotic one. Moreover, several reports of natural animal infection have been noted in the country. In this paper, we review how CL due to L. tropica is present around the Mediterranean basin, with a particular focus on the Moroccan foci, and try to unveil the existence of a zoonotic transmission in the area. A deep understanding of the transmission patterns of L. tropica in the region is necessary for the implementation of effective control measures and a better assessment of the disease burden. In addition, the complexity of this neglected disease requires more fundamental and applied research to unveil potential factors involved in its spread.Cutaneous leishmaniasis (CL) due to  Leishmania and transmitted through the bite of female sand flies. The disease is distributed worldwide and characterized by 3 main clinical forms: cutaneous leishmaniasis (CL), visceral leishmaniasis (VL), and mucocutaneous leishmaniasis (MCL) [Leishmania major, Leishmania aethiopica, and Leishmania infantum are responsible for zoonotic CL . Leishmania tropica is mainly considered responsible for anthroponotic CL. Healed CL due to L. tropica can sometimes be followed by Leishmaniasis recidivans (LR), usually appearing as new lesions around the primary healed ones and characterized by a progressive expansion [The development and spread of infectious diseases are influenced by humans, animals, and the environment, and around 61% of human infections are zoonotic. Zoonotic diseases are constantly emerging and expanding, and new transmission patterns are being uncovered [is (MCL) . In the xpansion .L. tropica has a wide geographical distribution in Greece, Turkey, and North African countries. CL due to L. tropica is characterized by its highest incidence in Morocco where it has been expanding since the 1990s [L. tropica cases [L. tropica could rise without the involvement of an anthropogenic spread [L. tropica is still debated. Several potential host species are now considered reservoirs in Algeria, Tunisia, Palestine, and Israel [L. tropica around the Mediterranean basin and Morocco and identify research gaps that need to be addressed.Around the Mediterranean basin, he 1990s . These lca cases . The emeca cases . As climc spread . The antd Israel –10. HoweLeishmania tropica” were combined to the following keywords: “Zoonotic,” “Reservoir,” “Animal host,” “Mediterranean,” “Morocco,” “Vector,” “Epidemiology,” and “Distribution.” Articles in English and French were browsed. No year limitation was assigned. Several combinations of these keywords were entered, and articles were selected from titles and abstract matching the reviews’ aim. Relevant articles on the epidemiology of CL due to L. tropica in the Mediterranean basin, the distribution of L. tropica’s vector, and the animal hosts of L. tropica in the same area were included in the manuscript.A review of the literature was conducted through articles obtained via PubMed, Web of Science, and Google Scholar databases. Several sets of keywords were used for this search. The terms “Cutaneous leishmaniasis” and “L. tropica has widely been reported across the Mediterranean basin. Syria is historically endemic to CL due to L. tropica for centuries and had previously the highest burden for this form of leishmaniasis in the region [L. tropica [L. tropica in the region. The largest portion of cases originates from the Mediterranean southern coast of Turkey [L. tropica is endemic, although its cycle of transmission is not fully elucidated in the region, and in Egypt, the first L. tropica CL cases were reported in 2009 from a classical L. major focus in North Sinai [CL due to e region . These le region ,11. Two  tropica ,13. A sith Sinai .L. tropica is characterized by the widest geographic distribution in the country, as 43% of CL cases  were due to L. tropica between 2008 and 2017 [L. tropica cases have increasingly been reported from different regions of the country. In 1991, an ecoepidemiological study unveiled a vast L. tropica focus in central and southern areas of the country such as Guelmim, Agadir, and Essaouira [L. tropica even spread to Settat Province that was nonendemic to CL. From 2007 to 2012, 553 cases were reported from this province, most of which originated from the same locality (Lbrouj) [L. tropica’s presence was confirmed in semirural localities of Toundout and Agdz, which are separated by the Atlas Mountains, north and south of Ouarzazate. This was the first report of L. tropica in the south of the Atlas Mountains, since the species is endemic to central arid and semiarid regions and northwest to the Atlas Mountains [L. tropica foci have spread to several regions of Morocco including those that previously reported CL caused by L. major or L. infantum. Recently, a survey carried out in Casablanca highlighted the possible introduction of L. tropica to urban areas [In North Africa, Morocco has the highest burden of this form of CL. Currently, CL due to and 2017 . It is pand 2017 . During ssaouira . In 1996ssaouira . In 2006(Lbrouj) . More reountains . Over thL. tropica infection were notified [Leishmania killicki (syn. L. tropica) are continuously reported in Algeria, Libya, and Tunisia [In Algeria and Libya, only sporadic cases of notified . Neverth Tunisia . It has  Tunisia .L. tropica has only been reported in Greece, where it was recorded for the first time in 1984 and occurs sporadically. Nevertheless, the proximity of certain European countries to endemic neighbors alongside the anthroponotic character generally recognized for L. tropica, as well as the presence of the vector Phlebotomus sergenti poses a major risk of introduction of CL due to L. tropica in south Europe [Finally, in southern Europe, CL due to h Europe .Phlebotomus (Paraphlebotomus) sergenti is the proven main vector of L. tropica. It is characterized by a wide geographical distribution in the Mediterranean basin [L. tropica in Algeria, Israel, Morocco, and Tunisia. Moreover, it is suspected to be a vector in each of Greece, Libya, Palestine, Syria, and Turkey [an basin ,28. It id Turkey .P. sergenti is still the main proven vector of L. tropica, Phlebotomus arabicus and Phlebotomus similis have also been studied for their vectorial capacities for L. tropica. Indeed, P. arabicus has been proven as a potential vector in northern Israel, and the widespread presence of P. similis (classified in the same subgenus as P. sergenti) in the island of Crete makes it a likely vector of L. tropica in the region [L. tropica was detected in other species of Phlebotomus sand flies, mainly Phlebotomus tobbi and Phlebotomus perfiliewi that could be potential vectors of L. tropica [Leishmania species may be isolated from P. sergenti [L. tropica in experimental conditions [Leishmania parasite originating from the same area must be proven [Even though e region . Moreove tropica . Althougsergenti , it is snditions . Howevere proven .P. sergenti is wider than the parasite it harbors. This species is distributed in CL endemic and nonendemic Mediterranean countries, where it is widely spread across semiarid regions. In Morocco, the first detection of L. tropica in P. sergenti was reported in 1991 from the first documented focus of Tanant [P. sergenti infection with L. tropica was reported in central and the northern parts of the country [P. sergenti and Phlebotomus papatasi are the most prevalent sand fly species, and they equally share the territory, although their abundance and density are under the influence of ecological factors. For instance, P. sergenti’s density is significant in arid and Saharan zones, whereas P. papatasi is capable of well adapting to different arid zones [P. papatasi, P. sergenti was found up to 1,400-m altitude in Al-Haouz region of the High Atlas mountains [P. sergenti was distinguished by an anthropophilic character and was mainly collected in domestic and peridomestic habitats [Interestingly, the geographic distribution of f Tanant . Since t country –38. In M country . P. sergid zones . In contountains . In thishabitats .P. sergenti was previously commonly found in mild to high altitudes and was described as a “mountainous” species, in Morocco, it was found in areas up to 2,000 m [Although  2,000 m . In turn 2,000 m .L. tropica have been recorded in Europe, P. sergenti is present, and even high infection rates of L. tropica were detected in 21 sand fly species in Greek refugee camps [P. sergenti in the Iberian Peninsula and the identification of one common mitochondrial lineage in both Morocco and southwestern Europe, in addition to the presence of L. tropica in northern parts of Morocco [L. tropica [P. sergenti poses a risk of introduction of L. tropica to the region. Even though sand flies are known to travel short distances, the anthroponotic character of L. tropica poses a threat for the emergence of cases, since a heavy traffic of potentially infected tourists or migrants exists between this region and the endemic North African foci.Interestingly, while no autochthonous cases of CL due to ee camps . This spee camps , identifee camps , as wellee camps . Its preee camps . Moreove Morocco , as well tropica . SoutherL. tropica is disputed. Some potential mammalian hosts have been investigated and could contribute to the transmission of the disease  and Massoutiera mzabi were suggested as possible reservoir hosts of the L. killicki (syn. tropica) [L. tropica genotype was detected in Gundis, humans, and P. sergenti within the same southeastern Tunisian L. tropica foci [Gerbillus pyramidum floweri was found to be infected by L. tropica [In other North African countries, CL cases due to smission . In Tunitropica) ,9. Inter tropica .L. tropica hosts. No apparent lesion was detected in black rats (Rattus rattus) experimentally infected with L. tropica, making them a potential asymptomatic host for the parasite. Moreover, these black rats originate from an endemic L. tropica focus in Turkey and were found to be infective for the sand fly vector [L. tropica was reported from small wild rodents of Turkey [Procavia capensis) are now considered as a reservoir [L. tropica. Wild canids such as golden jackals and red foxes were infected by L. tropica and thus may transmit this species from infected hyraxes to naive hyrax populations or even humans [L. tropica was detected in cats in Turkey and in wild rodents and bats in Ethiopia [L. tropica was notified from Jerusalem [Throughout the Mediterranean basin, several rodent species were investigated for their capacity to be y vector . This imf Turkey . In Israeservoir . Higher eservoir . The reseservoir . These rn humans . In addiEthiopia –61, and erusalem . TherefoL. tropica in the Mediterranean basin. Rock hyraxes are now considered as reservoirs in Palestine and Israel where both anthroponotic and zoonotic transmission cycles coexist. In Morocco, the relative variety of animal hosts that were shown to be infected by L. tropica suggests that its transmission cycle may also be zoonotic  as well as artificial rocky habitats (rock piles). This affinity could be explained by the fact that rock hyraxes live in these rocky habitats. P. arabicus, incriminated in L. tropica transmission, also shares the same habitat preferences as P. sergenti. Anthropogenic landscape changes, particularly the increase of rock piles, resulted in an increase of L. tropica CL cases in the region [Environmental changes to habitats of hosts and vectors affect the potential manifestation of zoonotic diseases. In Israel, e region .L. tropica in urbanized settings [L. tropica foci is linked to human travel, since the country’s road and railway infrastructure system has evolved in the recent years, increasing population movements [L. tropica in Morocco  can offer an accurate and user-friendly alternative for remote areas, as they can instantly give results while the patient is still in consultation. A recent study conducted in Morocco compared the efficacy of an RDT to standard tests to diagnose patients from various endemic areas. Even though the test had a relatively low sensitivity, it still could be used in isolated localities and would be an important addition to clinical CL management in Morocco. However, further improvements need to be done in terms of cost .L. tropica’s transmission patterns in the Mediterranean context highlights the need of a multidisciplinary approach to control the disease. The current Coronavirus Disease 2019 (COVID-19) pandemic spotlights the threat for humans posed by potential outbreaks of zoonotic infectious diseases, making it necessary to watch closely emerging infectious diseases and fully understand the risk factors involved in their spread. In Morocco, a zoonotic spillover from one of the potential animal hosts of L. tropica is very likely, and it may enhance the disease’s spread or may even impact the clinical manifestations, leading to more severe ones. Some gaps are still hampering the fight against CL due to L. tropica in our country. For instance, the reporting system still needs improvements, especially in remote endemic areas. Moreover, not enough investigations were done regarding a potential animal reservoir, reducing the efficiency of the preventive measures taken by health instances. This added to a gap in applying protective measures in endemic localities by local populations seriously affects the disease burden. Finally, the general public is still not entirely aware of the disease, making it urgent to instigate awareness campaigns throughout the country.The complexity of L. tropica in Morocco, to collect as much data as possible from endemic areas to understand its transmission pattern. It is also important to consider the training of professionals in endemic remote areas in order to improve the health reporting system. Moreover, a diverse group of experts should be involved in decision-making as part of One Health collaboration such as epidemiologists, physicians, entomologists, veterinarians, and researchers. The complexity of Leishmaniases and infectious diseases in general recalls for integrating a multidisciplinary approach to control their spread. In the case of CL due to L. tropica, this global approach could lead to not only better apprehend the disease dynamics, but also initiate the establishment of more efficient preventive techniques that take into consideration every aspect of the disease. To promote a coordinated response to zoonotic public health challenges, efficient One Health actions should take into consideration 3 types of factors: (i) individual factors such as education and training; (ii) organizational factors including communication and organizational structures; and (iii) network factors including network structures, leadership, and management. The implementation of these factors is crucial to fostering effective collaborations and multisectorial response to health concerns [Therefore, the existence of concomitant anthroponotic and zoonotic transmission cycles should not be overlooked. If a zoonotic transmission cycle is proven, a targeted control strategy of the animal reservoirs should be set up. It is therefore important to closely monitor the evolution of CL due to concerns . The firL. tropica, but also to design new diagnostic and prevention tools. When implementing control programs, local sand fly epidemiology and behavior and multiple local transmission pathways must be taken into consideration. Finally, local populations should play an active role in the control process, and the control methods used must be sustainable.Nonendemic Mediterranean countries must be prepared for the next potential CL outbreak. Thus, it is urgent to prioritize research in basic and applied science not only to complete the knowledge of the transmission cycles due to Leishmania tropica is endemic to many countries of the Mediterranean basin where it is widely distributed.Cutaneous leishmaniasis (CL) due to L. tropica in the Mediterranean basin, and some were incriminated as reservoirs.Several mammalian hosts were found to be naturally infected by Phlebotomus sergenti is present in nonendemic north Mediterranean regions, suggesting the possible introduction of L. tropica in these areas.A potential concomitant zoonotic transmission in Morocco should not be overlooked.One Health initiatives are key to implementing efficient control programs.10.1016/S0140-6736(18)31204-2Burza S, Croft SL, Boelaert M. Leishmaniasis. Lancet. 2018;392:951–970. doi: 10.1016/j.actatropica.2013.12.018Chaara D, Haouas N, Dedet JP, Babba H, Pratlong F. Leishmaniases in Maghreb: An endemic neglected disease. Acta Trop. 2014;132:80–93. doi: 10.1051/parasite/2014014Aoun K, Bouratbine A. Cutaneous Leishmaniasis in North Africa: a review. Parasite. 2014;21:14. doi: 10.1016/j.pt.2016.04.007Han BA, Kramer AM, Drake JM. Global Patterns of Zoonotic Disease in Mammals. Trends Parasitol. 2016;32:565–577. doi: 10.2807/ese.15.10.19505-enReady PD. Leishmaniasis emergence in Europe. Euro Surveill. 2010;15(10):29–39. doi:


Brugia malayi is known to be zoonotically important because it can be transmitted from animals  to humans or from humans to humans through mosquito vectors. This study was conducted to explore the fauna associated with Malayan filariasis transmission in Sedang village, Suak Tapeh District, Banyuasin Regency, South Sumatra Province, Indonesia.Mansonia spp. was observed. The presence of microfilaria was confirmed under a microscope in night blood samples of inhabitants and cats. The presence of infective larvae (L3) of B. malayi was identified microscopically and based on the polymerase chain reaction method in female Mansonia mosquitoes.A cross-sectional research design with an observational and analytical approach was applied in this study, and it was conducted in May 2018. Mosquitoes were collected twice using human bait both inside and outside the house from 6:00 p.m. to 6:00 a.m. The presence of competitors, predators, and reservoir hosts in the areas of five breeding habitats of Mansonia uniformis was the dominant mosquito, and the predominant competitor was Mansonia annulifera. Dragonflies, as predators were found in two breeding habitats and fish were found in one breeding habitat. The L3 of B. malayi were not identified in the mosquitoes, and the microfilariae of B. malayi were not found in the blood samples of inhabitants and cats.A total of 12 mosquito species were found, among which Mansonia mosquito population was abundant in Banyuasin Regency, the mosquito was not confirmed as an intermediate host of B. malayi, and the cat was not confirmed as a reservoir of B. malayi in the location.Although  Wuchereria bancrofti, Brugia malayi, and Brugia timori and is a major health problem in several tropical and subtropical countries. In 2004, an estimated 120 million people suffered from filariasis in 73 filariasis-endemic countries with an estimated 1.3 billion people in the filariasis transmission area ) of mosquitoes collected using the landing method . Cats were found in every breeding habitat area and can act as reservoir hosts of the nocturnal subperiodic type of B. malayi  for insect growth and development, and in breeding habitat 2, the maximum number of B. malayi in female Mansonia mosquitoes. This finding indicates that filarial larvae were not detected in all the captured mosquito species.We found negative results in the microscopic examination and the molecular PCR method conducted to identify the L3 of Mansonia spp. growth, the presence of other mosquitoes as competitors, the presence of other insects or animals as predators, and the presence of animals as reservoir hosts.Epidemiologically, the transmission of lymphatic filariasis involves several complex factors, such as the presence of lymphatic filarial worms as agents of disease, humans as hosts, adult mosquitoes as vectors, and physical, biological, and social environmental factors. The environment is highly influential on the distribution and transmission chain of filariasis cases. The biological environment involved in the transmission chain of filariasis includes the presence of aquatic plants as a place of Mansonia mosquitoes was found to be abundant in this study, no B. malayi L3 were identified. Therefore, Mansonia mosquitoes were not confirmed as an intermediate host of B. malayi in this study. The microfilariae of B. malayi were also not found in cat blood samples in this study; therefore, cats were not confirmed as the reservoir of B. malayi in this location. Similarly, a study conducted in 2006 in Sungai Rengit village, Banyuasin Regency, South Sumatra [B. malayi L3 was not identified in the mosquitoes. Conversely, a study conducted in Narathiwat Province, South Thailand [B. malayi microfilariae in cat blood samples.Although the population of  Sumatra , and a s Sumatra , reporteThailand , and anoThailand  reportedThe development of insects, including mosquitoes, is influenced by two factors, that is, the internal factors from the insects themselves and the external factors that are present in the surrounding environment where the insects live. High or low insect populations at any given time are influenced by these two factors . All vecMansonia spp. in the breeding habitats with optimum temperature. Biological environment includes all the living things that are found around humans, that is, flora and fauna, which can act as a chain of filariasis transmission. Mansonia spp. are associated with swamps and large rivers on the edge of the forest or in the forest. Larvae and pupae with their siphon are attached to the roots or branches of aquatic plants such as E. crassipes, Nymphaea (water lily), and I. aquatica [Mansonia spp. In Pistiastratiotes and E. crassipes, the aerenchyma  is larger in size than that in Mimosa pudica and Azolla pinnata and can store more oxygen. The root tissues of M. pudica and A. piñata are more rigid and can inhibit perforation by the siphon larvae of Mansonia spp. The root structure of P. stratiotes and E. crassipes can reduce the effectiveness of natural predators from mosquito larvae in ponds. A. pinnata is believed to have secondary metabolites that are dangerous to Mansonia spp. larvae [Geographically, the mosquito is a cosmopolitan insect with a widespread distribution in the tropical and subtropical regions, and the changes in the environment influence the activity of the insect, which has an impact on its diversity and distribution . Mosquitaquatica . The veg. larvae .B. malayi are generally found around rivers, forests, swamps, along rivers, or other water bodies overgrown with aquatic plants. Areas with different flora have different disease patterns. Swampy or muddy environment and bushes around houses are suitable breeding habitats for the Malayan filariasis vector (Mansonia spp.), and there exists a relationship between the presence of aquatic plants and the incidence of filariasis. Therefore, people living in endemic areas in houses with aquatic plants could have a risk of transmission of filariasis [Endemic areas of lariasis .M. uniformis was the dominant mosquito, and the predominant competitor was Mansonia annulifera. The study results also indicated the presence of dragonflies as predators in two breeding sites and the presence of fish in one breeding habitat. Furthermore, despite the abundant population of Mansonia mosquitoes in Banyuasin Regency, the mosquito was not confirmed as an intermediate host of B. malayi, and the cat was also not confirmed as a reservoir of B. malayi in the location.This study found 12 mosquito species, among which BM: Designed the study. SRU and SH: Conducted the field survey. EE: Collected mosquito sample. All authors drafted, revised, read, and approved the final manuscript.


Rhipicephalus sanguineus group encompasses at least 12 validated species of Palearctic and Afrotropical hard ticks, which are relevant in veterinary medicine and public health. The taxonomy of R. sanguineus s.s., has been particularly intensely debated, due to its wide geographic distribution, morphological variants, parasite-host associations, and its capacity and vectorial competence for the transmission of several pathogens. By sequencing mitochondrial markers, it was possible to identify the existence of multiple lineages, among which the Tropical and the Temperate lineages stand out, particularly in America. However, the northern limit between these lineages is not clear due to the lack of extensive sampling across Mexico. For this reason, the aim of the present study was to determine the genetic diversity and structure of the R. sanguineus group in Mexico and to compare it with the populations reported in the Americas, in order to propose the northern limit of the R. sanguineus Tropical lineage and the potential regions of sympatry with R. sanguineus s.s. The findings of this study now confirm the presence of R. sanguineus s.s. in Mexico, showing a subtle genetic structure and high genetic diversity throughout its distribution in the Americas. In contrast, the Tropical lineage seems to be genetically less diverse in its overall distribution in the Americas. The genetic diversity of these two independent lineages could have important epidemiological implications in the transmission of tick pathogens.The  Rhipicephalus sanguineus group encompasses at least 12 validated species  of Palearctic and/or Afrotropical hard ticks widely distributed worldwide by several historical colonization processes, which exhibit a high relevance in veterinary medicine and public health due to direct damage caused by infestation in the hosts and by the transmission of pathogens  and CMT-30 [MZ618826]), four sequences from the US , and two from France (MH630343 and MH630344). The second group was formed by two sequences from Brazil (MF477856 and KX533942), Argentina (KU498302 and MT152825), and Chile (MW509950 and KX632155), as well as the single sequence from Uruguay (GU553084) [Finally, with the Nei’s distances, the group of U553084) .R. sanguineus group from the Americas. A total of 156 (77.3%) corresponded to the Tropical lineage and the remaining 46 records (22.7%) were from R. sanguineus s.s. The Tropical lineage is widely distributed across the Neotropical Region; meanwhile, R. sanguineus s.s. is restricted to North America, including the US and the US-Mexico border, and is also present in the South American Cone, where it is restricted to Argentina, Chile, and Uruguay , the cytochrome b (cytB), or the second internal transcribed spacer of the nuclear ribosomal gene cluster (ITS2) to further characterise the subpopulations [Specifically, this gene fragment revealed subtle genetic differentiation across the distribution of both lineages. Regarding ulations ,33,34.R. sanguineus s.s. were lower than those from the Tropical Lineage, when these two lineages were compared with the haplotype network, R. sanguineus s.s. seems to be more diverse than the Tropical lineage, with six haplotypes of the total 10. This was also supported by the genetic distances, where a subtle genetic structure could be shown within this species in North and South America. This obvious genetic diversity could be due to an isolation-by-distance effect or by two independent colonization events, that may have been influenced by multiple factors related to radiation across several new hosts and climate niche partitioning, as was recently proposed by Bakkes et al. [Although the numbers of available sequences from s et al. . HoweverR. sanguineus s.s., the use of the 16S rDNA gene fragment seems to be less informative for the Tropical lineage, containing only four haplotypes and without an obvious genetic structuring. However, it is important to note that in this case, a single haplotype (H2) concentrates most of the diversity and could be inferred as the differentiation centre, since all mentioned countries are included in this haplotype. This has been proposed for other biological systems, with some unique haplotypes from South America (Colombia and Brazil) as tip haplotypes [Amblyomma maculatum within the US [Contrary to the information obtained with plotypes ,36,37. Tn the US .R. sanguineus group to transmit, for example, E. canis in Brazil [This differentiation might be epidemiologically relevant due to variability in the vectorial capacity and the competence of some of the populations of the n Brazil .R. sanguineus s.s. and that of the Tropical lineage. In the Northern limit, the two sympatric locations for both lineages are highly crowded border crossings that allow a constant exchange between the populations of R. sanguineus of the US and Mexico through the movement of domestic animals.Based on the biogeographic regionalisation, no correlation could be evidenced between the presence of R. sanguineus s.s. and Tropical linage only share the Chacoan province in Argentina. In this context, the use of the distribution proposed in this study must be applied with caution until future efforts allow us to complete the distribution analysis in this area.On the other hand, in South America, where a more complex biogeographic regionalisation is found, due to the complexity of the biota and fauna, R. sanguineus s.s. corresponds to the previously known “Temperate Lineage”, containing a subtle genetic structure and high genetic diversity throughout its distribution in the Americas. In contrast, the 16S rDNA region seems to be less informative for inferring the genetic diversity of the Tropical lineage in its overall distribution in the Americas. Yet caution is warranted, since the analysis of the 16S rDNA gene fragment does not permit us to untangle the phylogenetic taxonomy of this complex group formed by other Palearctic and/or Afrotropical species, such as R. turanicus, R. sanguineus Southeast Lineage, R. guilhoni, R. leporis, and R. camicasi. To conclude, the findings of this study confirm that R. sanguineus group in the Americas, showing different biogeographical distributions. Finally, the genetic diversity shown by these two independent lineages could have important epidemiological implications in the transmission of tick pathogens.We also now propose the first fine scale map with the boundaries of these two lineages of the Rickettsia Surveillance’, we received brown dog ticks (R. sanguineus group) from the National Network of Public Health Laboratories of the Mexican states of Aguascalientes, Baja California, Campeche, Chihuahua, Coahuila, Guanajuato, Guerrero, Hidalgo, Morelos, Oaxaca, Puebla, Sonora, Veracruz, and Yucatan. Additionally, we analysed several ticks from previous tick-borne surveillance studies carried out in Chihuahua, Sonora, and Veracruz [As a part of the ‘National Program for Veracruz ,39,40. WVeracruz : the Nea® tube and cut into small pieces using a sterile scalpel blade for each specimen to avoid cross contamination between tick samples. The samples were then macerated with a 1.5-pellet pestle.All ticks were fixed and preserved in 70% ethanol and morphologically identified with an Olympus SZX7 stereoscopic microscope, using the specialised taxonomic keys of Keirans and Litwak , Direcci®, implementing the “Purification of Total DNA from Animal Tissues (Spin-Column Protocol)”. For molecular identification of the tick species, a segment of approximately 400 bp of the mitochondrial 16S rDNA gene was amplified in all samples, using the primers 16S + 1/16S − 1 and the cycling conditions reported by Norris et al. [® Green Master Mix, 2X Promega Corporation , the pair of primers (100 ng each), 6.5 μL nuclease-free water, and 30 ng DNA in a final volume of 25 μL. Negative (reaction mix without the DNA template) and positive  controls were included. The PCR products were resolved in 2% agarose gels using SmartGlow™ Pre-Stain of Accuris Instruments®  and visualised by UV-transillumination. Purified amplification products were submitted for sequencing at Macrogen Inc., Seoul, Korea.The DNA extraction was carried out using the Qiagen “DNeasy Blood & Tissue Kit”s et al. . The reahttps://blast.ncbi.nlm.nih.gov/Blast.cgi?PROGRAM=blastn&PAGE_TYPE=BlastSearch&LINK_LOC=blasthome). Recovered sequences were edited using FinchTV 1.5 , conducting a visual inspection. The final sequences were compared to each other to identify inter/intra species variation, using the BLASTn tool from NCBI (16S rDNA structure, using the Q-INS-i algorithm in the MAFFT version. The set of sequences included all available sequences of R. sanguineus s.l. from the Americas in GenBank, with the sequences of the neotype of R. sanguineus from France. Following the recommendations of Guglielmone et al. [Rhipicephalus sanguineus group, R. turanicus, R. guilhoni, R. leporis, R. camicasi, and R. sanguineus tropical lineage from the Australasian, Oriental, and Afrotropical regions according to Chitimia–Dobler et al. [R. sanguineus s.l. clade III [according to Nava et al. [Rhipicephalus microplus, as an outgroup. The complete list of these sequences and their geographical distribution are presented in We then performed multiple alignments according to the secondary structure of the r et al. , Hornok r et al. , and Slar et al. . AdditioTwo phylogenetic analyses were performed, and the best substitution model was evaluated differentially, following the recommendations by Colunga-Salas and Hernández–Canchola . The firmrbayes models and the greedy scheme search [The second approach was performed using MrBayes 3.2 . To calce search ,53 were e search .R. sanguineus Tropical Lineage and R. sanguineus s.s. from the Nearctic and Neotropical regions, without the outgroup. To provide a more detailed account of the genetic diversity and to detect possible populations and subpopulations in the R. sanguineus Tropical Lineage and R. sanguineus s.s., an ordination PCoA of the eigenvalue scores was performed in PAST 4.04 [For all following analyses, we only used the multiple alignment of the AST 4.04 , based oAST 4.04 .adegenet R package [Haplotype networks with mutational step estimations were constructed using the median-joining network in POPART 1.7 . Additio package , conside