Tengchong County was one of the counties located at the China-Myanmar border with high malaria incidence in the previous decades. As the pilot county for malaria elimination at the border area, Tengchong County is aiming to be the first county to achieve malaria elimination goal. A cross-sectional entomological survey was carried out to evaluate the feasibility of elimination approach and assess the receptivity of malaria reintroduction.Light traps associated with live baits were used to investigate the abundance of adult mosquitoes in nine villages in Tengchong County. Light traps were set to collect adult mosquitoes in both human houses and cowsheds from dusk till dawn in each site.Anopheles mosquitoes were collected from May to December in two villages. Of the mosquitoes were captured, 24.2% were in human houses and 75.8% in cowsheds. The peak of abundance occurred in July for An. sinensis and in September-October for An. minimus (s.l.) Ten Anopheles species were collected, the most prevalent being An. sinensis (50.3%), An. peditaeniatus (31.6%) and An. minimus (s.l.) (15.8%), contributing to 97.6% of the sample. Potential breeding sites were also investigated and a total of 407 larvae were collected, with An. sinensis (50.1%) and An. minimus (s.l.) (46.2%) as predominant species. Ponds and rice fields were the two preferred breeding sites for Anopheles mosquitoes; however, the difference between the number of adults and larvae captured suggest other breeding sites might exist. Both An. sinensis and An. minimus (s.l.) were found zoophilic with human blood index as 0.21 and 0.26, respectively. No Plasmodium positive Anopheles specimens were found by PCR among 4,000 trapped mosquitoes.A total of 4948 adult An. sinensis, associated to rice fields, is particularly worrying. Sustained entomological surveillance is strongly suggested even after malaria elimination certification.Although no indigenous malaria cases have been reported in Tengchong County since 2013, there is still a risk from the presence of vectors in the context of human population movements from neighboring malaria endemic areas. The presence of  Malaria is the deadliest vector-borne disease in tropical and subtropical areas, with a number of confirmed cases estimated at 216 million with 445,000 deaths in 2016 . HoweverTengchong County (TCC) is located in the southwest of Yunnan Province at the China-Myanmar border Fig. . Because2 with a population of 6.68 million inhabitants and an international borderline with Myanmar of 148 km . Mountains cover 84% of the territory with a maximum elevation of 3780 m  around the selected villages were investigated for larvae. The hand dipper sampling method was used to collect larvae  [28S rDNA sequences were run to identify the sibling species of the An. minimus, An. culicifacies and An. fluviatilis complexes and An. maculatus group [All potential terbody) . The morus group –24.Anopheles species were aggregated to analyze seasonal fluctuations. The resting behavior and breeding preference of adult mosquitoes were explored by analyzing the adult and larval composition in each study place. The adult population density for each Anopheles species was calculated as the number of females per trap per night (f/t/n). The overall (pooled) Anopheles density was calculated by summing captured individuals of all Anopheles species. Species richness was measured by the number of species and the indices described below. Generally, species diversity is an indicator of the wellbeing of an ecosystem [Monthly abundance data of each cosystem . Simpsoncosystem . The larThe indices were calculated as follows:Simpson’s diversity index: pi×ln piShannon-Wiener’s index: H =  −∑ pi is the fraction of a species which belongs to the i-th species and N is the number of species (pi = Ni/N).where H/ ln SEvenness index: E = H is the Shannon-Wiener’s diversity index and S is the total number of species observed in a given place.where Plasmodium spp. test while the abdomen was used for blood-meal identification. DNA extraction was conducted with QIAamp DNA Mini Kit  according to the supplier. PCR tests for blood source identification were conducted as previously described [Captured mosquitoes were dissected into different segments under light microscope. The head and thorax were separated for escribed –29. PrimMosquitoes were collected by six collectors using an aspirator in different locations, including human houses and cowsheds Fig. . The colhttp://www.gadm.org). The relief model was prepared using SRTM 90m digital elevation data v4.1 [Administrative spatial data were obtained from the GADM database of Global Administrative Areas (ata v4.1 . Land coata v4.1 . Data weThe diversity of species measured by the Simpson’s diversity index (D) and the Shannon-Wiener’s index (H), both for human houses and cowsheds, was at the highest in Man Lve and Nong Ling at the beginning of the fall, i.e. September-October, although the trend was already visible in August Table . The divAn. sinensis , An. minimus (s.l.)  and to a lower extent An. peditaeniatus . Three other species were collected, i.e. An. splendidus , An. culicifacies (s.l.)  and An. aconitus , but in very limited numbers. Anopheles sinensis was mostly present from June to August with a peak in July but remained present until November  was less prevalent but displayed a bimodal curve with two peaks in July and September. Anopheles peditaeniatus was recorded only from July to October with a maximum plateau in August and September. The survey of mosquito prevalence in cowsheds yielded a slightly different pattern  and An. peditaeniatus, but with different ratios than in human houses. Anopheles sinensis was still the most prevalent , followed by An. peditaeniatus  and An. minimus (s.l.) . Three more species were detected in cowsheds at a lower extent: An. splendidus , An. culicifacies (s.l.)  and An. argyropus . The main three species were recorded for the same period as in human houses. An. sinensis was mostly present from June to September with a peak in July; An. peditaeniatus was recorded from July to October, with a peak in August, and An. minimus (s.l.) displayed the same bimodal curve with peaks in July and September  were not found in Man Lve.The collection of adult mosquitoes in the village of Man Lve Fig.  was condern Fig. . The firber Fig. . Anopheln = 479) was in the same range as in Man Lve, with the same three main species, i.e. An. sinensis , An. peditaeniatus , and An. minimus (s.l.) . Unlike in houses in Man Lve, An. peditaeniatus was the second most prevalent species. The main difference was the population size in cowsheds: 22 in Man Lve and 112 in Nong Ling , although less than in Man Lve. The ratio between cowsheds and houses in Nong Ling was only 2.4 (1169/479). The most prevalent species in cowsheds was An. peditaeniatus  followed by An. sinensis  and An. minimus (s.l.)   displayed a plateau covering September and October. Beside these three dominant species, four more species were also collected such as An. culicifacies (s.l.) , An. liangshanensis , An. maculatus (s.l.) and An. splendidus  in human houses. Anopheles aconitus and An. argyropus, rare in Man Lve, were not found in Nong Ling.Adult mosquitoes were collected in the village of Nong Ling Fig.  over the69) Fig. . The bimAn. sinensis , An. peditaeniatus  and An. minimus (s.l.) . They contributed for 97.7% of the total Anopheles mosquitoes collected. An. sinensis was the predominant species in both human houses  and cowsheds . However, An. minimus (s.l.) was the second largest mosquito species in human houses . The PCR analysis of the 647 specimens of An. minimus (s.l.) indicated that 64.8% (419/647) were An. harrisoni (former An. minimus species C [An. minimus (former An. minimus species A) , while 79.9% were isolated from cowsheds (686/859) ; Yan Si (n = 123); Man Lve (n = 184); and Nong Ling (n = 306). The most frequent species in the 9 villages were also An. sinensis , An. minimus (s.l.)  and An. peditaeniatus . Anopheles sinensis was found in all the sites investigated and An. minimus (s.l.) in all but one site, Man Duo  and 35.Plasmodium parasite was detected in any captured mosquitoes (Table An. sinensis and 194 were An. minimus (s.l.) With respect to An. sinensis, 88 individuals (87.1%) were parous while 180 (92.8%) An. minimus (s.l.) mosquitoes were also found parous.No es Table . Out of An. sinensis was found more zoophilic (27.3%) than An. minimus (s.l.) (10.7%). Anopheles minimus (s.l.) displayed more mixed blood meals, either animal/animal (13.1%) or animal/human (14.3%) than An. sinensis  (Table An. sinensis and An. minimus (s.l.) are 0.21 (28/132) and 0.26 (44/168), respectively.A total of 300 blood samples from trapped mosquitoes were tested. y) Table . The humAnopheles species were identified among the fourth instar larvae and pupae collected , i.e. An. sinensis, An. minimus (s.l.), An. culicifacies (s.l.) and An. peditaeniatus. Anopheles sinensis  and An. minimus (s.l.)  were the predominant species , pool, ditch and rice field  is one of the major malaria endemic counties in Yunnan Province with both smission , 14, 18.smission . After 2Anopheles species or complexes were previously recorded as predominant malaria vector in TCC, i.e. An. minimus (s.l.), An. dirus (s.l.), An. sinensis and An. liangshanensis (syn. An. kunmingensis) [P. falciparum malaria with a local transmission in TCC at high altitude (> 1700 m), due to its greater susceptibility to P. falciparum compared to P. vivax [An. liangshanensis populations in line with the number of local P. falciparum malaria cases [Anopheles dirus was found neither at the adult nor larval stage in this study. This indicates that the population density of An. dirus has decreased and might now play a negligible role in malaria transmission. Similar results were reported in neighboring counties [An. dirus initially present as the primary malaria vector has disappeared [An. dirus might not only be due to vector control activities (particularly the use of LLIN/ITN) but also to the destruction of breeding sites such as forests to develop plantations of cash crops.Four ngensis) , 44. TheP. vivax . Howeveria cases , 46. Anocounties  and in Hcounties  where Anappeared , 43. TheAn. sinensis was found to be the predominant species in both human houses and cowsheds. This differs significantly from previous reports from neighboring counties [An. minimus (s.l.) was the predominant taxon. This is particularly important because An. sinensis displays specific traits making it a potential threat for malaria elimination. First, An. sinensis is associated with rice fields [An. sinensis has been reported as resistant to insecticides such as pyrethroids and Malathion [An. sinensis displayed a similar tropism to humans as An. minimus (s.l.). These traits, combined with the predominant abundance of An. sinensis in TCC, are major concerns for the success and sustainability of malaria elimination. The exact role of An. sinensis in malaria transmission in TCC and, more widely in Yunnan Province, should then be thoroughly investigated. Moreover, regular movements of populations across the China-Myanmar border, owing to the existence of endemic malaria in Myanmar [An. sinensis. This threat must thus be further assessed and modeled while scenarios of risk management must be developed. Furthermore, An. minimus (s.l.), another malaria vector in TCC, was consistently found in this study with two peaks of density during the year. A large part of the An. minimus (s.l.) population (64.8%) belonged to An. harrisoni. This could explain the lower local malaria transmission in TCC since An. harrisoni was reported to be more exophagic and zoophilic than An. minimus, its sibling species [An. harrisoni was also reported to be more adaptable to the environmental changes and flexible in its trophic behavior than An. minimus [An. minimus and An. harrisoni was reported to be a consequence of vector control measures in Southeast Asia [An. minimus (s.l.) found in this study may have resulted from vector control measures, as well as environmental changes increasing the proportions of An. harrisoni versus An. minimus. Unfortunately, PCR techniques for identification of the Anopheles complexes were not used in previous routine surveillance and there is thus a lack of detailed information about the initial distribution of An. minimus (s.l.) [An. minimus (s.l.) This indicates that more detailed integrative analyses should be conducted to better understand the mechanisms involved in the dynamic of vector populations. A closer attention should also be brought to PCR species identification, an approach to be implemented in routine surveillance at malaria elimination stage and post-elimination stage. Despite the decrease of population density of An. minimus (s.l.) in TCC, the ecological behavior such as resting or blood-seeking behavior was found to be similar as previously described prior to population decrease [An. sinensis and An. minimus (s.l.) in potentiating malaria transmission should not be ignored. Owing to its zoophilic diet preference and considering the lack of competence for transmission of malaria parasites, An. peditaeniatus was not previously reported as a malaria vector [P. falciparum in one specimen of An. peditaeniatus was recently confirmed by ELISA in Indonesia [An. peditaeniatus was the second largest population of adult mosquitoes found in this study, further investigation should thus be conducted to monitor the risk of malaria transmission by this species in TCC. Another aspect to consider is the discrepancy between the number of adults and larvae of An. peditaeniatus captured. Only three fouth-instar larvae of An. peditaeniatus were identified out of 199 Anopheles larvae indicating a very low prevalence in all the breeding sites investigated. Conversely, there is a high prevalence of adults in cowsheds. This suggests that the actual breeding sites of this species were most likely missed. It is therefore essential to investigate thoroughly all possible breeding sites for An. peditaeniatus.In this study, counties , where Ae fields  and theralathion , 49. Thi Myanmar , 6, incr species , 50–52.  minimus , 53–55, ast Asia . Comprehast Asia . The coms (s.l.) . It is tdecrease . Furthera vector . Howeverndonesia . Since AAn. peditaeniatus was found in large number in only three localities out of nine and at low level in two more. The two main vectors, An. sinensis and An. minimus (s.l.), were present in all localities and in all but one, respectively. Nevertheless, the main difference is the species richness and the number of individuals between human houses and cowsheds, which might be linked to Anopheles blood preference or more favorable living conditions. Although some predominant Anopheles species are known as zoophilic, a reduction of livestock may favor malaria re-emergence as some species are quite ubiquitous like An. sinensis [Environmental factors should also be considered when conducting entomological survey. sinensis . Furthersinensis .Plasmodium-infected mosquito was found, the high parous rate of An. sinensis (87.1%) and An. minimus (s.l.) (92.8%) suggests a high daily survival probability [An. minimus complex and An. sinensis across TCC and the border areas; (iii) Evaluation of the length of the possible transmission season for P. falciparum and P. vivax; and (iv) Evaluation of the vectorial capacity of An. minimus, An. harrisoni and An. sinensis.Although no bability . ContinuAnopheles populations, and the risk posed by secondary vectors and insecticide resistance, further efforts should be devoted to surveillance, monitoring and development of scenarios for timely response to imported malaria cases. A specific attention should be paid to local environments and variation of vector prevalence when developing scenarios. Large-scale analysis might not be accurate and reliable enough. Precise actions from both local CDC and national program at this border area will be essential for the success of sustainable malaria elimination.TCC was granted the malaria elimination certificate by Yunnan provincial authorities in 2016. However, considering the increasing mobility of the local populations, the border location with Myanmar, the positive vector competence of local


Lutjanus analis and 19 of Lutjanus jocu, of which 46 isopods from 3 different species were collected, i.e. Rocinela signata, Cymothoa excisa, and Excorallana richardsoni. The species R. siganata and E. richardsoni are reported for the first time in L. jocu. A significant relationship between the parasite R. signata and the size of the fish L. jocu was also observed. The isopod C. excisa is considered an incidental finding in L. analis. To the authors' knowledge, this is the first report of this species in fish from Brazil. The three species of isopods are new occurrences in the State of Sergipe, northeast region of the country. An additional morphological characteristic observed in the dorsal setae of pleotelson in specimens of E. richardsoni was that one end of this structure wws bifid. This information contributes to the current body of knowledge of the morphology of this particular species.Parasitic isopod species are poorly known in the northeastern coast of Brazil. In this sense, this study presents novel records of Isopoda of the families Aegidae, Cymothoidae, and Corallanidae. A total 69 specimens of Lutjanus analis  - and the “Dentão” - Lutjanus jocu . These two species are important fish resources in the State of Sergipe, northeast region of the country [Lutjanidae is a family of cosmopolitan fish that occur most often in regions of tropical and subtropical seas of Australia, Africa, North America and South America. In Brazil, several species of this family are widely explored by artisanal fishermen. Because of their generalistic carnivorous behavior, they play an important role in the ecological control of other populations in reef environments of coral reefs. On the Brazilian tropical coast, 15 species of fish belonging to Lutjanidae have been reported including the “Cióba” - country . BesidesCuvier, 128 - and  country –14. HoweFor this survey, fish were purchased monthly according to the supply and availability at the Municipal Fishing Terminal of Aracaju, SE, Brazil (10°54′17′′S 37°2′56′′W) between March 2015 and October 2016 directly from artisanal fishermen working along the northeastern coast of Brazil in the Western Atlantic. These fish were shipped to our laboratory and identified at the species level according to the taxonomic keys published by , and Sil L. analis, and 19 specimens of L. jocu were analyzed, of which 13.8% were parasitized by at least one species of isopod. We found a total of 46 specimens of parasites which belonged to the families Aegidae, Corallanidae and Cymothoidae. Cymothoa excisa Perty, 1833 , Rocinela signata Schioedte & Meinert, 1879 and Excorallana richardsoni Lemos de Castro, 1960 were the species of parasites from these three families that were found in the fish examined. R. signata was the species of greater prevalence in both L. analis and L. jocu, followed by E. richardsoni and C. excisa . The parasite was found in males and females of two host species: L. analis - 4 isopods in two male specimens and in L. jocu - 6 isopods in 5 male specimens and 1 isopod in 1 female specimen. A male of L. jocu had 2 species of isopods in its gills: R. signata and E. richarsoni.The speciesThe following are the morphological characters and taxonomy of the specimens found in the fish examined:Aegidae Leach, 1815Rocinela Leach, 1818Rocinela signata Schioedte & Meinert, 1879 Location: Northeastern Coast, Aracaju, SE, Brazil.Cymothoidae Leach, 1818Cymothoa Fabricius, 1793Cymothoa excisa Perty, 1830 Location: Northeastern Coast, Aracaju, SE, Brazil.Corallanidae Hansen, 1890Excorallana Stebbing, 1904Excorallana richardsoni Lemos de Castro, 1960  including Cavalcanti et al. [ R. signata in L. synagris with 10% of parasitized fish and a mean intensity of 1.25. The parasite has a distribution from Central America to South America, and has been recorded by Hermida et al. [ L. analis, corroborating the registry in the same host of the present study, these authors showed a prevalence of 3.3% and an average intensity of 1.00. Similar results were obtained in the present study, The most recent record of parasitism was published by Cardoso et al. [ Pseudupeneus maculatus Bloch, 1793 with a prevalence of 8.3%, an average intensity of 1.1, and a variation of 1 or 2 parasites per fish. In the present survey, a lower prevalence of isopods was observed in the fish examined. The gill chamber was the preferred site of infestation for the parasite R. signata. The fish L. jocu is reported here as a new host for R. signata.Lima et al.  recordedi et al.  that reca et al.  in L. ano et al.  in the c R. signata had the same number of antenulla segments and antenna flagella as described by Brusca and Iverson [ R. signata is an isopod frequently found in fish in northeast Brazil. As other isopods, R. signata have a negative impact on fish's health. It induces slow growth and respiratory problems and cause injury to the host especially when large numbers of parasites are observed. In cases of severe parasitic infestation, opportunistic secondary fungal and bacterial infections may occur, which is most observed in confinement situations. However, because they are coastal fish, it was not possible to measure such disturbances, since parasite intensity and host immunity are also a determining factor for the occurrence of infections in aquatic populations.Specimens of Iverson  with 4 a Iverson  and Herm Iverson , R. signCymothoa excisa is a species of isopod that has low specificity, and in the world is already known occurring in several vertebrates, in fishes has been reported in 6 different families of fish including Lutjanidae [ Micropogonias furnieri (Sciaenidae) . The occurrence of C. excisa in L. analis is in agreement with previous studies published by Weinstein [ Lutjanus, in the Yucatan Peninsula, Mexico and Panama, where it was also recorded in three other species of Lutjanidae: Lutjanus synagris, L. mahogoni and Ocyurus chrysurus [ L. analis in Brazil. Although they are protandritic animals, becoming functional males in one part of the cycle and later becoming functional females [tjanidae , 27. In tjanidae  in Microeinstein  and Bunkeinstein . These ahrysurus . However females , accordi females , isopods L. analis and L. jocu which are both reef fish species, still have some differences regarding habitat: L. analis has preference for sandy bottoms with vegetation, estuaries, and regions near mangroves whereas L. jocu prefers coastal waters, particularly estuaries, and occasionally rivers. C. excisa was found only in L. analis, in this sense, Weinstein [Allen  states teinstein  explain  E. richardsoni was also found only in male L. jocu. Barriga and Briones [ Excorallana in the east coast of Mexico; 7 species were identified: E. acuticauda Miers, 1881; E. delaneyi Stone and Heard, 1989; E. oculata Hansen, 1890; E. subtilis Hansen, 1890; E. sexticornis Richardson, 1901; E. tricornis tricornis Hansen, 1890; and Excorallana sp. Luque et al. (2013) reported Excorallana sp. on the body surface of Ageneiosus inermis Linnaeus, 1766, and C. excisa in the buccal cavity of Micropogonias furnieri Desmarest, 1823. To date, these are the only records of these two parasites of fish in Brazil. Koening [ Excorallana oculata along the northeast coast of the country from the State Amapá to the State of Espírito Santo.The species Briones  reported Koening  document E. richardsoni is distributed in the States of Amapá, Pará, Maranhão, Ceará, Rio Grande do Norte, Pernambuco, Alagoas, Espírito Santo, and Rio de Janeiro. The first report of this isopod in the country was published by Koening and Coelho [In Brazil,d Coelho  in the Sd Coelho  in the S E. richardsoni are in agreement with those presented by Silva and Souza-Filho [ Excorallana have bifid setae with mid-apical setae in the dorsal pleotelson region (In the present study, the morphological characteristics ofza-Filho . Menziesza-Filho  describen region . In our n region , arrow.  R. signata presented the highest infection rates in L. analis and L. jocu. The presence of E. richardsoni and R. signata in the State of Sergipe, which lies on the northeast Brazil's Atlantic coast, may have occurred due to the migratory habit of the fish since this ectoparasites has been previously recorded in waters of the State Alagoas, northeastern region of the country, thus increasing its geographical distribution. Although the bifid end of the dorsal setae of the pleotelson is not a definitive morphological character used for the taxonomic identification of E. richardsoni, this particular feature was observed in all specimens examined in this study. The present survey presents the first report of three species of isopods in the state of Sergipe, the first record of Cymothoa excisa in Lutjanus analis in the country. We describe Lutjanus jocu as a novel host for Rocinela signata and Excorallana richardsoni.In conclusion, low levels of isopod infection were found in the fish species studied.


Schistosoma. Worldwide, an estimated 250 million people are infected with these parasites with the majority of cases occurring in sub-Saharan Africa. Within Asia, three species of Schistosoma cause disease. Schistosoma japonicum is the most prevalent, followed by S. mekongi and S. malayensis. All three species are zoonotic, which causes concern for their control, as successful elimination not only requires management of the human definitive host, but also the animal reservoir hosts. With regard to Asian schistosomiasis, most of the published research has focused on S. japonicum with comparatively little attention paid to S. mekongi and even less focus on S. malayensis. In this review, we examine the three Asian schistosomes and their current status in their endemic countries: Cambodia, Lao People’s Democratic Republic, Myanmar, and Thailand (S. mekongi); Malaysia ; and Indonesia, People’s Republic of China, and the Philippines (S. japonicum). Prospects for control that could potentially lead to elimination are highlighted as these can inform researchers and disease control managers in other schistosomiasis-endemic areas, particularly in Africa and the Americas.Schistosomiasis is an infectious disease caused by helminth parasites of the genus  Schistosoma. Six species of schistosomes infect humans: Schistosoma mansoni , S. haematobium , S. intercalatum and S. guineensis , S. japonicum (Asia), and S. mekongi , and previously Thailand whose current status is transmission interruption) [S. malayensis, which is thought to be closely related to S. mekongi, is endemic in Malaysia [S. japonicum, S. mekongi, and S. malayensis, which cause intestinal schistosomiasis. The three Asian schistosomes are all zoonotic, whereas the remaining species infecting humans are generally considered human-only parasites, with some notable exceptions [Schistosomiasis is a parasitic disease caused by blood flukes of the genus ruption) ,2,3. A sMalaysia . In thisceptions ,7,8,9,10S. japonicum eggs in a mummy [Schistosomiasis has a long history in Asia with the first descriptions and reports of the disease in modern times appearing in the early 1900s, although it is thought to have been endemic for at least 400 years earlier in Japan, and at least 2200 years ago in the People’s Republic of China (P.R. China) after the discovery of  a mummy ,12,13,14 a mummy . Current a mummy . Conside a mummy . Old cha a mummy .The aim of this review is to provide an overview of the current status of schistosomiasis in Asia, with a particular focus on endemic countries in the region and the unique challenges they face. Our review also aimed to identify current knowledge gaps and future research needs as the affected countries move toward the ultimate goal of control and elimination of this persistent and debilitating disease.Schistosoma is a group of parasitic blood flukes, or flatworms, of the class Trematoda. Unique amongst the trematode class, schistosomes have separate sexes as adults, whereas all other trematodes are hermaphrodites. The Asian schistosomes discussed in this review are considered zoonotic, unlike schistosome species occurring elsewhere, which are largely human-only excepting some hybrid forms in Africa [S. mansoni infecting non-human primates in Africa and the Caribbean, and rats in Guadeloupe and Brazil [S. japonicum is the most cosmopolitan, with 46 mammalian definitive hosts identified thus far, whereas S. mekongi has been found in dogs, and S. malayensis in rodents, specifically Rattus muelleri [S. mekongi, but, to date, no natural infections have been identified in these hosts [The genus n Africa ,9,10,18,d Brazil ,8. S. jamuelleri ,20,21,22se hosts . Morpholse hosts .The schistosome lifecycle is complex with an intermediate molluscan host, definitive host, and seven lifecycle stages involving both asexual and sexual reproductive phases . An in-dS. japonicum is the most prevalent of the Asian schistosomes. It is endemic in P.R. China, the Philippines, and small foci occur in Indonesia. There are 46 known mammalian definitive hosts of S. japonicum, although water buffalo and cattle have previously been shown to be the major reservoirs of infection [S. japonicum was first identified in Japan in 1901, whereas the last new human case was recorded there in 1977. S. japonicum parasites in P.R. China and the Philippines have distinct genetic differences, resulting from geographic isolation over time. In general, the strain of S. japonicum in P.R. China is more virulent than the parasite in the Philippines; additional genetically variant geographic isolates are known to be present in both countries [nfection ,28,29. Sountries ,31,32.S. mekongi was first identified in 1857 [S. japonicum, S. mekongi differs in a number of characteristics that indicate it is a distinct species. These differences include the morphology of the testis and ovary in adult worms [S. mekongi are smaller and more round than those of S. japonicum , and studies have found Oncomelania spp. snails to be refractory to infection with S. mekongi [Apart from human infection,  mekongi .S. malayensis is zoonotic and is primarily a parasite of the rodent R. muelleri , and the third ‘chronic’ stage ,37. As tThe initial disease phase begins as a skin rash caused by an immune reaction to the penetrating cercariae , I. AfteA number of diagnostics are available, including coproparasitological examination (CopE), as well as molecular and immunological diagnostics. CopE methods rely on direct detection and visualization of parasite eggs in feces and include the Kato-Katz (KK) thick smear procedure, which is a mainstay of control programs due to the relative ease of performing the test at low cost, although it lacks sensitivity in low-intensity infections ,47 and FSensitive and specific diagnostic procedures are required to monitor the success or failure of schistosomiasis control programs as well as to determine whether control efforts have resulted in elimination. However, the most sensitive diagnostics, involving molecular or immunological techniques, can be expensive and require specialized facilities and equipment and trained personnel to perform the procedures ,76.S. japonicum and S. mekongi [Laboratory studies and clinical trials have shown that praziquantel, a pyrazinoisoquinoline derivative, is a safe and highly efficacious oral drug that is active against all schistosome species, although it is less active against juvenile schistosomes compared with adult worms and eggs ,79,80,81 mekongi ,78. Praz mekongi . A singl mekongi .Treatment with praziquantel does not prevent reinfection  and is tDue to the requirement of an intermediate host snail, schistosomiasis is a focal disease, occurring in areas where snail habitats and susceptible transmitting snails are present. This means that village-level prevalence can be very high, whereas country and province prevalence can be low. Demographic factors such as age, sex, and occupation are strongly associated with risk of infection ,88. OpenSnail habitats generally occur in still or slow moving water bodies such as streams, lakes, dammed waterways, and rice fields. The susceptible snails also have a preference for vegetation and snail control measures can include the removal of this vegetation.S. japonicum hosts, bovines, particularly water buffalo, are considered the most important for transmission due to the high levels of schistosome eggs they excrete into the environment [S. japonicum infections, and hence are considered to be the most important reservoir hosts [S. japonicum in P.R. China include goats, pigs, and dogs due to their close contact with humans and water [Of the 46 known ironment ,90 and tir hosts ,89,90,91ir hosts ,93. Othend water .S. japonicum [S. japonicum in water buffalo  in the Philippines has been reported to be as high as 80%, particularly in agricultural areas, and water buffalo are thought to be a major reservoir [S. japonicum for much longer. Although studies suggested that water buffalo exhibit some age-acquired resistance to infection and self-cure [In Indonesia, 13 mammalian species, mostly wild animals , but also cattle, water buffalo, horses, and dogs, have been identified as susceptible hosts for aponicum . The preeservoir . In bothelf-cure ,97,98, telf-cure .S. japonicum and S. mekongi, bovines could act as reservoir hosts of S. mekongi but, to date, this has not been demonstrated. A range of potential animal hosts have been examined for S. mekongi, but currently dogs are the only animal species that have been confirmed as natural hosts of this species [Due to the close genetic relationship between  species .S. japonicum, with the exception of P.R. China, which has practiced both chemotherapy of bovines and removal of the animals, facilitated by mechanization of agriculture  [S. mekongi has been highlighted as a priority. This has been proposed as part of community-led initiatives to eliminate schistosomiasis that combine deworming with water, sanitation, and hygiene interventions: community-led school, water, sanitation, and hygiene (CL-SWASH) activities [In regards to control, little has been done to target definitive hosts of ractors) ,100,101.ractors) . In Lao tivities .Animal vaccines against schistosomiasis have been developed and used in controlled trials ,106,107.S. japonicum are amphibious and belong to the genus Oncomelania, with species dependent on geographical location. Studies investigating the susceptibility of snails from different geographic locations to cercariae from disparate locations have produced mixed results, indicating a certain amount of genetic drift of the parasite [Oncomelania hupensis nosphora and O. hupensis formosana, are still present [The intermediate snail hosts of parasite ,112. Alt present .Oncomelania hupensis have been identified based on morphological and molecular characteristics [O. h. guangxiensis and O. h. tangi [O. h. hupensis and O. h. robertsoni as the dominant sub-species [O. h. hupensis is the most widely distributed [S. japonicum infection rates in Oncomelania snails in endemic regions of P.R. China in the last decade. One study in the marshland regions along the Yangtze River [In P.R. China, four sub-species of the intermediate host eristics ,115,116,h. tangi  leaving -species . Of thestributed . Few stuze River  examinedze River . An earlze River . Accordize River . Howeverze River . As snaize River ,120.S. japonicum is O. h. quadrasi. It is amphibious but prefers an aquatic environment, such as wet soil surfaces, swamps, rice fields, ponds, and stream banks, thus making chemical snail control difficult due to the risk of contaminating the water or food source [S. japonicum infection in the snail intermediate hosts in the Philippines is poorly understood. One study, conducted in Samar province, found a mean infection rate of 1.09% across 147 sites with higher infection among snails located in irrigated compared to rain-fed villages [In the Philippines, the sole intermediate host snail for d source . The curvillages . More revillages .S. japonicum is O. h. lindoensis, which is located focally around Lake Lindu. In 2011, the prevalence of infected O. h. lindoensis snails in Lindu Valley was 3.6% and 4.0% in Napu Valley, although the prevalence has fluctuated between 0 and 13.4% in Napu Valley and between 0% and 9.1% in Lindu Valley since 2005 [In Indonesia, the intermediate snail host of nce 2005 . Prevalence 2005 .S. mekongi are Neotricula spp., and endemic areas are closely associated with the Mekong Delta where these snails occur. The prevalence of infected snails in Lao PDR was quite low, 0.01% on the Mekong Islands [Neotricula aperta snail density in Thailand decreased between 2005 and 2011 in the downstream area of the Nam Theun 2 hydroelectric dam, which began operation in 2010 [Oncomelania snails in low-land areas was initially seen in P.R. China after the building of the Three Gorges Dam [The intermediate hosts of  Islands  and 0.22 Islands . Neotric in 2010 . A similrges Dam . HoweverControl of intermediate host snails is an important aspect of schistosomiasis control programs, particularly as the stage of the lifecycle occurring in snails is asexual and involves an exponential increase in parasite numbers. Snail control measures previously implemented in Asia have involved environmental modification and chemical mollusciciding. The snails live in vegetation around rivers and lakes and removal of this vegetation can lead to removal of the snails themselves. This method was used with great success in Japan in combination with mollusciciding. In Japan, the most common method of environmental modification was the use of concreting canals where the snails lived . This meMollusciciding has also been used in P.R. China, Indonesia, and the Philippines, although to a limited degree in the latter two countries. Environmental contamination with chemical molluscicides is an important issue, and a number of previously used compounds have been abandoned due to these concerns and as a result of the damage they cause to the environment. In the schistosomiasis foci in Indonesia, the snail habitats occur close to the Lore Lindu National Park, which precludes the use of molluscicides  and enviNeotricula spp., the intermediate host snails of S. mekongi, exhibit a different ecological niche than those transmitting S. japonicum in that, rather than being present in marshlands and rice fields, these snails are primarily found in shallow areas of rivers (particularly the Mekong River) and tributaries. Thus, snail control for S. mekongi has largely been deemed infeasible [feasible , althougfeasible .S. japonicum transmission are related to distance to a snail habitat or those that influence snail habitats, such as the building of dams. The majority of S. japonicum-endemic zones are within 1 km of water bodies such as rivers, lakes, or wetlands [The majority of environmental factors associated with wetlands ,132. Envwetlands ,134, seawetlands .S. japonicum, and might account for 95% of the snail habitats [Oncomelania snails survive best at areas of low elevation—one of the potential environmental factors associated with high prevalence of snails in marshy areas. A study on snail habitats in mountainous and hilly areas identified a maximum elevation of 2300 m above sea level for snail survival [Oncomelania snails are amphibious; yet, require water to survive. A distance of ≤1000 m from the nearest stream was found to be ideal for snail habitats [In P.R. China, endemic areas occupy three different geographical landscapes: (1) marshland and lake areas, (2) mountainous and hilly areas, and (3) water network areas. Of these, marshland and lake areas are characteristic of the major endemic foci for habitats ,87. Oncosurvival . The samhabitats .As the marshy and lake areas are categorized by the presence of water bodies, more areas are available for the snails to exist. The area of these landscapes, which cover the four provinces of Hunan, Jiangxi, Anhui, and Hubei, is vast, complicating snail control in these locations . MountaiO. h. hupensis as well as O. h. robertsioni and O. h. guangxiensis [Oncomelania spp. snails. The decrease in flooding events has decreased the density of snails in some areas, although in others, the density appears unchanged, or is on the increase [Local epidemic outbreaks and the geographic distribution of snail hosts are heavily influenced by flooding events caused by the Yangtze River as they facilitate snail dispersion to new localities such as rivers, lakes, and wetlands ,137. Largxiensis . The Thrincrease .S. japonicum infection: 80% in Mindanao, 18% in Visayas, and 2% in Luzon [S. japonicum infection prevalence differently in the three main regions of the Philippines [O. h. quadrasi include flood plains, forests, and swamps, whereas man-made habitats resulting from agricultural development are thought to be important habitats . These snails are generally found on banks but also occur in shallow water (depth <20 cm) [O. h. quadrasi snails prefer areas shaded by vegetation where the temperature is relatively stable and cool.In the Philippines, more than 3000 bodies of water are thought to be infested with snails susceptible to in Luzon . Endemicin Luzon ,82. Envilippines . As is tlippines . Some dilippines . A confo <20 cm) . O. h. qS. japonicum in snails from this area ranged from 0 to 13.4% in the Lindu Valley and 0 to 9.1% in the Napu Valley, although human prevalence remained <1% as of 2006 [Endemic regions of Indonesia are located in marshland areas around Lake Lindu and Napu and Bada valleys. Prevalence of  of 2006 .S. mekongi transmission occurs in the Mekong Delta. In Khong and Mounlapamok districts in Lao PDR, 202 villages are situated along the Mekong River with 114 currently or previously endemic for schistosomiasis. The only villages with zero prevalence for S. mekongi are in parts of the river where the riverbed is sandy, which is not conducive to the intermediate host snail, or those villages that are more than 6 km away from the river [he river .Limited information is available regarding risk factors and snail intermediate hosts in Myanmar . The curS. japonicum. Infection with S. japonicum is strongly associated with age, sex, and occupational exposure. Males aged 40 years and above who engage in fishing, farming, and herding are at greatest risk of infection [In P.R. China, transmission usually occurs across two distinct seasons , coincidnfection . DefecatIn the Philippines, there is no distinct dry season on the main endemic islands of Leyte and Samar and the province of Mindanao. Hence, transmission is not as variable by season as in P.R. China and occurs all year round . The mosRattus have been suggested as the primary source of transmission, with a peak prevalence of 20% found in one endemic village [R. exulans, R. hoffmani, R. chysocomus rallus, R. marmosurus, and R. celebensis [Whereas 13 species of mammalian hosts have been identified in Indonesia, limited research has been undertaken on their involvement in transmission. Rodents of the genus  village . Primarylebensis . Reservalebensis ,93. As iS. mekongi is matched with the lifecycle of the snail. During times of high water levels (2–3 m), the majority of available snails are young, while in times of low water , the snails have matured to adults capable of carrying the infection. Peak transmission of S. mekongi in Cambodia occurs between February and April coinciding with peak water use for fishing [The transmission season for  fishing ; whereasIn Malaysia, the wet season differs between the southwest where the monsoon season is May–October, and the northeast where the monsoon season runs from November to March, and the typhoon season occurs from April to November. It is therefore difficult in the absence of yearly surveys to pinpoint when transmission in Malaysia might peak, but it is certainly influenced by rainfall brought by the monsoons and typhoons.Preventive chemotherapy with praziquantel has been the mainstay of schistosomiasis morbidity control and, in addition to targeting mammalian and snail hosts mentioned earlier, efforts to control transmission have also included programs aimed at improving WASH and health education. Until relatively recently, the role of WASH in schistosomiasis control was limited ,147,148,Health education is important not only for educating the public on risk reduction measures and changing behavior, but has also been found to facilitate diagnosis, surveillance, and treatment . ThroughS. japonicum, there is the rationale for the development and deployment of a transmission-blocking anti-S. japonicum vaccine targeting bovines [Research is ongoing for the development of schistosomiasis vaccines targeting humans and, unlike the African schistosomes, the zoonotic nature of the Asian schistosomes allow for the targeting of animal hosts. With bovines, particularly water buffalo/carabao confirmed as the major reservoirs of  bovines ,166,167. bovines  and clusNotably, most prevalence estimates reported are based on microscopic detection of schistosome eggs in stool samples, usually using the KK thick smear technique ,168,169.S. japonicum in P.R. China and the Philippines occurred around the same time in the early 1900s [The first reports of ly 1900s ,172 and ly 1900s .S. japonicum was once found in Japan, the last reported human case was in 1977, and elimination of schistosomiasis was declared in 1996 [Oncomelania nosphora, is still present [As indicated in the species name,  in 1996 . There w in 1996 . Due to  present . In addiOncomelania snails bred, began in 1938 and was the primary method used [Environmental modification in the form of concreting canals, where the hod used . In addihod used . Land ushod used . EducatiS. japonicum is predominantly found in areas along the middle and upper reaches of the Yangtze River Valley in the southern part of the country where the climate and environment are highly suitable for the propagation of Oncomelania snails. Endemic regions are concentrated in the lake regions and in the mountainous region in the western part of the country [S. japonicum was endemic in 12 provinces, but due to political will and sustained efforts, primarily through snail control and preventive chemotherapy, five provinces have achieved transmission interruption. Of the remaining seven provinces, four  achieved transmission control  by 2014, whereas Anhui, Jiangxi, and Hunan are still in the infection control stage  [In P.R. China,  country . Until ranimals) ,175. In animals) . In 2016Control approaches in P.R. China have been extensive due to the strong commitment by the national government. Funds from a 10-year World Bank Loan Project (WBLP) implemented in the 1990s  were putS. japonicum is distributed throughout all three major island groups , although the majority of cases occur in Mindanao and the Visayas [S. japonicum is more widespread in the Visayas compared to Luzon and Mindanao, where it is more focal [In the Philippines,  Visayas ,180  in 1991, which was financially backed by a World Bank loan. The focus shifted to active case finding and mass drug administration with praziquantel; WASH interventions and snail control were included as additional measures. Drug coverage of the target population was reported to be 100% during the 1990s . However50 years ,183,184.50 years ,183.In the Philippines, niclosamide has been banned for use in snail control under the Clean Water Act . So, whiS. japonicum infection in animals by strengthening veterinary health teams in priority areas through capacity building and operational research and moving toward implementation of WASH programs, as outlined during the 17th Meeting of the Regional Program Review Group on Neglected Tropical Diseases in the Western Pacific (175). These efforts are laudable and it can be anticipated that WASH will feature more prominently in the Philippines than in the past [The success of P.R. China’s efforts since after the launch of the WBLP compared to the Philippines is largely attributable to the Chinese targeting both human and animals for chemotherapy as opposed to humans only in the Philippines, which ultimately proved to be far less effective . The Phithe past . Lessonsthe past ,96.S. japonicum has fluctuated over the last decade, prevalence tends to be higher in Napu Valley and overall appears to demonstrate an upward trend. Up to 2005, control efforts had decreased prevalence from 37% to 1% or less in Napu and Lindu valleys, but in the period from 2008 to 2011, prevalence varied between 0.3% and 4.8% in Napu and 0.8% to 3.2% in Lindu [In contrast to P.R. China and the Philippines, schistosomiasis in Indonesia is endemic in three comparatively small, isolated highland regions surrounding Lore Lindu National Park in central Sulawesi. These areas include marshes around Lake Lindu, particularly in the villages of Anca, Langko, Tomado, and Puro’o; Napu Valley  located in Lindu . When Bain Lindu .Schistosomiasis control strategies set by the National Objectives for Health (NOH) directive for 2011–2016 varied based on the degree of endemicity, but overall took a multifaceted approach. Control objectives included mass drug administration coverage of 85% of the entire population in high endemic areas, active selective treatment in moderate endemic areas, and passive selective treatment in low endemic areas. Preventive chemotherapy has been supplemented with strategies that also focused on treating domestic animals, snail control, health education, improving water, and sanitation, and monitoring and evaluation and capacity building  in an efS. japonicum and S. mekongi [n = 315), whereas 302 cases were identified between 2016 and 2018 [S. mekongi prevalence of 3.9% (n = 205) in the Bago Region of Myanmar [Myanmar has been previously thought to be non-endemic for schistosomiasis, although there have been some historical unconfirmed reports of the presence of both  mekongi . Recent  mekongi . Serologand 2018 . The WHOand 2018 . Recentl Myanmar ,197.A recent schistosomiasis outbreak occurred in Rakhine State with >400 confirmed cases and >800 suspected cases as of August 2018 . RakhineS. japonicum has been definitively diagnosed in Myanmar. Until the status of schistosomiasis is further clarified, ideally by molecular methods with which S. mekongi has already been confirmed, we rely on early reports that suggest both species are present [It is not immediately clear from published reports if  present ,197.The first case of schistosomiasis occurring in Cambodia was identified in 1968 in Eastern Cambodia and was seemingly confined to Vietnamese fishermen living in raft houses. Prevalence among children was higher (14–22%) than in adults (7–10%) . At presControl of schistosomiasis in Cambodia has been impacted by political unrest and upheaval through the 1970s and 1980s . It was S. mekongi, described as a S. japonicum-like infection, was reported in 1950 in Thailand [The first case of Thailand . This ‘pThailand . ThailanThailand .S. mekongi in 1957 in a patient who had been living in Paris for nine years, but had spent the first nine years of life in Lao PDR [S. japonicum-like egg. It was not until 1966 that a second case was identified in Lao PDR and, in 1969, an epidemiological survey determined a prevalence of 14.4% (n = 72) on Khong Island. As with S. japonicum, hepatosplenomegaly was associated with infection [Lao PDR followed Thailand in the identification of  Lao PDR ,199. Thinfection ,199,204.nfection .n = 994) in humans and 14.7% (n = 68) in dogs for S. mekongi [n = 29,583) of collected snails being positive [A study examining the prevalence of a range of parasites in islands of the Mekong in Lao PDR performed in 2011 determined a prevalence of 22.2%  [S. malayensis.Schistosomiasis resembling  autopsy . Previouingapore . A reviein 1988) ,207. No in 1988) . The strin 1988) , and likS. malayensis were identified in patients who were deceased with schistosomiasis identified in autopsies, although not as the reported cause of death [S. malayensis have been aboriginal Malaysians (Orang Asli) living in rural areas and patients were either previously or concurrently co-infected with other infectious diseases. The first case identified in a living patient occurred in Selangor State and the patient presented with hepatosplenomegaly [Until 1978, all cases of of death ,208. In nomegaly ,208.S. malayensis has only been recorded sporadically in humans, with few recent accounts of infected individuals. The most recent identification of infection in humans was reported in a pathology report of an individual in 2011 [In the meantime,  in 2011 . In this in 2011 .Multi-component, intersectoral, and integrated control approaches provide a promising path forward for the elimination of schistosomiasis in Asia. Behavioral changes that prevent infection, such as avoiding the practice of open defecation and contact of open freshwater bodies in endemic areas, are necessary. However, without accompanying infrastructure, such as toilets provided in WASH programs, these behaviors will continue. Combined with chemotherapy of both humans and reservoir hosts , snail control, animal vaccination, health education, and WASH targeting multiple points in the schistosome life cycle will significantly impact the prevalence and re-infection of schistosomiasis. The success achieved in P.R. China in schistosomiasis control and elimination is due to the commitment and support of the Chinese government, including the removal of water buffalo—a major reservoir host in P.R. China—from endemic areas, thus effectively removing them from the transmission cycle. Without such governmental support, sustaining control programs is difficult, particularly those that include more than preventive chemotherapy and require considerable resources to implement, such as required for CL-WASH. There is a niche role for health education that is missing from many schistosomiasis control programs, as knowledge remains limited about the parasite and which behaviors lead to infection in endemic populations.S. mekongi, particularly for S. malayensis, the number of human cases, and the role played by animal reservoirs in transmission. Thus, assessing the true importance of schistosomiasis in the countries where these two schistosome species are endemic is difficult.Limited data are available on prevalence for S. japonicum prevalence is conceivably underestimated, both in P.R. China and the Philippines, due to the lack of sensitive diagnostics used in control programs. While P.R. China closes in on elimination targets, the use of sensitive diagnostics will be important to determine whether elimination has indeed occurred and to prevent re-bounding infections after treatment and the cessation of the control program. A recent study in P.R. China found a human prevalence <1% by the MHT but 11% by qPCR [The true  by qPCR . AlthougFuture challenges for schistosomiasis control and elimination include climate change and the potential spread of the disease to new areas ,211,212.


Fungal infections affect over a billion people, with mortality rates estimated at 1–2 million per year . AdvanceGalleria mellonella, that is one of the most common insect invertebrate models used in biomedical research. G. mellonella is an insect with functional similarities to the innate immune response of mammals. In this review, the authors outline the immune response of G. mellonella with a focus on the cellular process of phagocytosis, nodulation, and encapsulation, as well as other responses, such as antimicrobial peptides (AMPs), melanization, clotting, free radical production, and primary immunization. The main advances in the G. mellonella model are also discussed, including the sequencing of its genome and the availability of transcriptomic data. Notably, recent results from transcriptomic analyses reveal that the immune system of G. mellonella can distinguish between specific classes of pathogens.Pereira et al.  present G. mellonella respond to infection by different fungal pathogens. For example, after infection by Candida albicans, the hemocytes of G. mellonella induce nodulation around C. albicans cells and this process is accompanied by early melanization. The immune response to Aspergillus fumigatus also results in the formation of melanized nodules and accumulation of antimicrobial peptides. Interestingly, infection with non-lethal inocula of C. albicans or A. fumigatus is enough to protect the larvae against a subsequent infection with lethal inocula. Interestingly, in the case of Cryptococcus neoformans, the infection in G. mellonella is not accompanied by early melanization due to the presence of the capsule and to the different cell wall composition. In the infection by C. neoformans, fungal cells are avidly phagocytosed by G. mellonella hemocytes, but can survive and replicate after internalization, behaving as intracellular pathogens similar to infection processes in mammals. Moreover, Singulani et al. [G. mellonella against other important fungal pathogens, including Paracoccidioides spp., Histoplasma capsulation, Blastomyces dermatitidis, Sporothrix spp, and Talaromyces marneffei.Extending the Pereira et al.  report, i et al.  highlighG. mellonella, this model creates the possibility of determining the efficacy of antifungal agents by evaluating both larval survival, as well as by monitoring the immune response associated with the infection process [G. mellonella model for the study of antifungal treatments. Many topics are discussed in this article, such as the assays to test relative toxicity in vivo, to identify the optimum combinations of antifungal drugs for resistant microorganisms, to study the mode of action of different classes of compounds, and to establish dosages prior to use in mammals. In another article, Cruz et al. [G. mellonella larvae infected with C. albicans, and reached the same efficacy as fluconazole. Among them, compound 2 (2-((2-(hexan-3-ylidene) hydrazino)-4-(4-methoxyphenyl)-thiazole) showed the most promising results and was identified as a potential candidate for further studies in murine models.Given that several fungal infections can be studied in  process . In this process  explore z et al.  show ineG. mellonella model has been widely adopted by many researchers around the world [G. mellonella for research, including age, size, availability of food, reared techniques, physical stress to transportation, incubation temperature, storage time, and possible residual levels of antibiotics and hormones. The authors suggest solutions on how to address these issues such as the pathological scoring system using automated real-time imaging, and developing of a standard well-characterized G. mellonella strain for research. While research and standardized procedures advance rapidly to make G. mellonella a robust and consistent infection model [Tenebrio molitor  as a potential model for the study of pathogenic fungi. This insect host has similar characteristics to G. mellonella, however its breeding in laboratory conditions is less laborious. These authors outline the first studies with T. molitor involving C. albicans, C. neoformans, Malassezia furfur and Fonsecaea pedrosi, and discuss the advantages of this model for the study of fungal virulence, mycotoxin effects, host immune responses, and the efficacy of antifungal agents.The he world ,20,21,22he world ,23,24. Ion model  present Caenorhabditis elegans is approached by Elkabti et al. [Candida spp. infections. A great focus is given to the development of new visualization techniques to enhance capturing infection processes within live worms, tracking of nematodes, and the automation of high throughput assays. Other interesting topics are also discussed, such as the recent studies on the microbiome of C. elegans in nature, and the use of this model to identify molecular mechanisms involved in the development of drug resistance. In another article, Rosowski et al. [In addition to insect model hosts, the nematode i et al.  who desci et al.  present Acanthamoeba spp., a free-living protozoa also used as a model host for fungal pathogens [To complete this special issue, Casadevall et al. raise important aspects in relation to athogens . The artOverall this special issue highlights the consensus regarding the alternative model hosts, such as amoebae, insects, worms, and zebrafish that are valuable and reliable tools to study fungal pathogenesis, but there are still several challenges and unexplored fields to be conquered.


AbstractHymenophyllaceae collected on Ambon and Seram islands, the Moluccas, Indonesia, during 1983–86. The collection is composed of forty-seven species and one variety belonging to seven genera. The dataset is deposited in GBIF and available at https://www.gbif.jp/ipt/resource?r=seram_hymen.Identifications are given for 713 specimens of  The flora of Seram and Ambon islands, the Moluccas, covering bryophytes, pteridophytes and seed plants, was investigated during field expeditions in 1983, 1984–85 and 1986. As the Moluccan islands, in particular Seram, have been explored very sparsely, the expeditions aimed to make general collections of the land plants in the area. More than 11,000 field numbers of vascular plant and 5,000 bryophyte specimens were collected, mainly from east, central and west Seram .PageBreakfying our collections. Hymenophyllaceae, were already published by M. Kato and his collaborators , Indonesia.Hymenophyllaceae.PageBreakAmbon and Seram islands, the Moluccas, Indonesia. The collection route map for the 1983 trip is given in Hymenophyllaceae, were collected in Seram and Ambon islands on the expeditions 1983–1986.Pteridophytes specimens, including those of Hymenophyllaceae remained unprocessed. After identification of the specimens by the authors, following the classification system by The pteridophyte flora of Ambon and Seram (Ceram) was comprehensively explored in the 1980s and was studied by M. Kato and his colleagues during 1985 and 2007. Most of the pteridophyte collections have already been studied, but the specimens of Hymenophyllaceae for Flora Malesiana, this taxonomically interesting family has been revised and the taxonomy of the species has been determined, including the identification of the collections cited here. A modern system, including information based on molecular systematics, was proposed by The field research was organised as a joint survey by the Botanical Gardens, the University of Tokyo and Herbarium Bogoriense, LIPI. Along with the work on the See C.apiifolia (C. Presl) Copel.47. Object name: A Specimen List of Hymenophyllaceae of Seram and Ambon collected on Indonesian-Japanese botanical expeditions 1983–1986Character encoding: UTF-8Metadata Language: EnglishPageBreakResource Language: EnglishType: OccurrenceSubtype: SpecimenData License: Creative Commons Attribution (CC-BY) 4.0Thesaurus: GBIF Dataset Subtype Vocabulary: https://rs.gbif.org/vocabulary/gbif/dataset_subtype.xml


I was saddened and quite shocked when I received the phone called informing me that Professor Zhengyi Wu had died. It was the evening of June 20th, 2013. I had just got off a plane from Tibet and turned on my mobile phone. Prof. Wu had been unwell for nearly a year. I knew that this would happen one day, but refused to believe it would really come to pass. A few days ago, some students and I were on a field trip in Tibet. Sitting in our jeep, I watched the mountains and roadside as we drove, and tried to identify the plants along the route. The mountain roads in Tibet seem to always make my students sleepy. I woke them up. I told them that I had heard this first when Prof. Wu was on a field trip to Tibet. During the trip, he always kept an eye on the surroundings outside the vehicle, observing plants and taking notes. At the end of each day, he would prepare a check-list of plants for the region through which he had travelled. Back home it was the middle of the night, and I was awake. My thoughts had turned to my relationship with Professor Wu , volume one, general overview  , written by Qing dynasty botanist Qiyong Wu, and The Illustrated Book of Japanese Botany. In middle school, he was fortunate to meet two Biology teachers: Mr. Shou Tang, his primary middle school Biology teacher, and Mr. Yao Tang, his middle school Biology teacher. They introduced him to fieldwork, and taught him to recognise plants and collect plant specimens. Profoundly inspired by his teachers, he decided to become a botanist. In 1933, he entered Tsinghua University in Beijing and received his BA in Biology four years later. By that time, the War of Resistance against Japanese Aggression had begun, and it was not possible to pursue research in botany. He moved, together with his University, to Kunming, Yunnan Province, which was home to the front during the war. He taught at National Southwestern Associated University and was a staff member at the Medicinal Plant Institute of the National Ministry of Education. During the war, teaching and research conditions were very poor. Regardless, Wu still tried his best to use his botanical knowledge to serve his country. Prof. Wu and other colleagues published a book called ICONES Plantarum Medicarum e libro Tien-Nan-Pen-Tsao Lanmaoano Tom 1 (滇南本草图谱) . At the time, medicine was badly needed but in extremely short supply. Their research, which identified medicinal plants around Kunming, was able to help. Twenty-six medicinal plants, including a new genus, Psammosilene, W. C. Wu et C. Y. Wu were described in the book, which included hand-drawn illustrations. Everything required for publication—writing, drawing and printing—all had to be done by themselves.The manuscript was finally completed, but I made one request. I asked Prof. Wu to write an autobiographical note for the book. He was happy to comply. One week later, Prof. Wu gave me his autobiographical note. When I read it, I was deeply moved. Prof. Wu was born in 1916. He developed an early interest in botany, inspired by his home garden and two books: Florae Republicae Popularis Sinicae (FRPS) in 1987, and 82 books made up of 54 volumes were published under his editorship . During the compilation of FRPS, he also chief edited another two great works: Flora Yunnanica and Flora Xizangica. In order that FRPS could be understood by non-Chinese speakers, Prof. Wu and Peter H. Raven became the co-chairs of the joint editorial committee. All 25 volumes of Flora of China were published in 2014, one year later Prof. Wu passed away unfortunately (Zhou and Sun, Since then, Prof. Wu started his scientific career which spanned over 70 years. Many articles have recorded his contributions to botanical studies already, here I only want to emphasise his contributions in plant taxonomy and phytogeography. Prof. Wu became the fourth editor in chief of the Editorial Committee of The Areal-Types of Seed Plants and Their Origin and Differentiation. He proposed about origin and differentiation of distribution pattern of many seed plants from China. This hypothesis became a big legacy to biogeograpical research. Many research projects were proposed based on Prof. Wu’s hypothesis (Zhou and Sun, By compiling the FRPS Prof. Wu and his colleagues answered the big question of how many plant species we have in China. His phytogeographcial research answered the next big question: “where are these plants growing”. His contribution to phytogeography has been recorded in the above-mentioned book We hope he will be remembered in the history of botany forever.


Mycobacterium houstonense belongs to the unnamed third biovariant complex of the Mycobacterium fortuitum group, which are sorbitol positive. To date, there have been few reports of human infection induced by M. houstonense worldwide.Historically M. houstonense and Escherichia coli. An implant bone plate had been embedded for internal fixation during surgery on the humeral fracture previously. A week later E. coli was isolated from the skin wound secretions. Cefoperazone-sulbactam was used for treatment for two weeks but the infection was not controlled, with a subsequent risk of deep wound infection. External fixation of the fracture was then performed instead of internal fixation. Ten days later, M. houstonense was isolated from new wound secretions. M. houstonense was identified by the molecular sequencing method. The TREK Diagnostic System was used to test the susceptibility to antibiotics by the microbroth dilution method. Levofloxacin and amikacin were used for treatment according to the results of the susceptibility test and the patient’s condition obviously improved.We describe the case of a 68-year-old man with surgical wound infection, following an open humeral fracture, caused by M. houstonense following open humeral fracture. The combination of levofloxacin and amikacin was effective in the treatment of M. houstonense infection.To the best of our knowledge, this is the first case in China of human surgical wound infection caused by  Mycobacterium fortuitum group, the Mycobacterium chelonae/abscessus group and the Mycobacterium smegmatis group [M. fortuitum group comprised M. fortuitum (formerly M. fortuitum biovar fortuitum), Mycobacterium peregrinum (formerly M. fortuitum biovar peregrinum) and the taxon known as the unnamed third biovariant complex. The M. fortuitum third biovariant complex mainly included sorbitol positive and sorbitol negative organisms, represented by Mycobacterium houstonense and Mycobacterium bonickei, respectively.Rapidly growing mycobacteria (RGM) can produce colonies that do not appear or slowly appear with pigmentation on various solid media within 7 days (most within 3 to 4 days), and contain mycobacterial acid and maintain amylase activity for 3 days or 2 weeks , 2. Tradis group . HistoriMycobacterium species: M. senegalense, M. mageritense, M. septicum, M. porcinum, M. neworleanense, M. brisbanense, M. houstonense and M. bonickei [At present, based on the evolution of 16S ribosomal gene (rDNA) sequencing, the unnamed third biovariant complex includes the bonickei , 4. AmonM. houstonense was first reported by the CDC (USA) and was isolated from the face wound of a patient who lived in Houston, Texas, so was named Mycobacterium houston [M. houstonense have been in the USA and Australia [M. houstonense in China. To the best of our knowledge this is the first report of M. houstonense in China. houston , 5. To dustralia . Here weA 68-year-old man had an open fracture of the right humerus due to a fall. The patient was sent to the hospital as an emergency case. The doctor performed debridement and suture of the patient’s wound. When all of the test indexes were normal, surgery of the humeral fracture was undertaken. Large bone defects in the middle and lower parts of the humerus were found during the operation. Fig.  After prEscherichia coli was isolated from the wound secretion by culture. E. coli isolates were multidrug resistant as determined by antimicrobial susceptibility testing using the disk diffusion test. The procedure and interpretation of the results of the antimicrobial susceptibility tests were conducted in accordance with the CLSI 2018 guidelines [A week later, wound secretions exuded through the original drainage tube. idelines . AntimicTwo weeks later, the drainage tube had been closed but yellowish cloudy secretions exuded on the lateral side of the arm incision. The doctors suspected that the deep wound was infected, therefore, re-debridement of the patient’s wound and external fixation of the fracture were performed Fig. .Fig. 3ExM. fortuitum group using an IVD-MALDIBIOTYPER . The isolated strain was identified as M. houstonense by sequencing analysis. Monoclonal colonies were scraped and genomic DNA of the isolate was extracted using a commercial kit . Primer design was based on the reports of Lane(1991) and CLSI MM18-A, and the primers for 16S rRNA PCR were as follows 27F: AGAGTTTGATMTGGCTCAG, 1492R: TACGGYTACCTTGTTACGACTT. The amplification conditions for PCR were based on those of previous reports [M. houstonense (GenBank accession no. NR_042913.1).Ten days later, another secretion from the wound was observed Fig. . RGM werM. houstonense by the microbroth dilution method. The antimicrobial drug sensitivity results were interpreted according to the CLSI M24 A2 guidelines [M. houstonense was only sensitive to levofloxacin, moxifloxacin and amikacin.The TREK Diagnostic System  was used to test the antimicrobial drug susceptibility of idelines . The resA daily intravenous drip of 0.3 g of levofloxacin and injection of 100 ml of sodium chloride, and injection of 0.2 g of amikacin and 250 ml of sodium chloride twice a day, were used for treatment. Three weeks later, the wound was healing well and no secretions were detected.M. houstonense in China. Reports of infection induced by M. houstonense are scarce throughout the world. The mechanism of infection with this organism remains unclear. Some studies have reported the isolation of M. houstonense from freshwater fish and fish products, especially retail frozen fish [M. houstonense. However, in this case, the patient had not recently been exposed to fish products. In the first report of M. houstonense in the USA, M. houstonense was isolated from a wound in the patient’s face [M. houstonense often inhabits water, soil and dust. Healthcare-associated outbreaks and community-acquired diseases of RGM are usually related to the pollution of water or soil [M. houstonense after an open fracture.To the best of our knowledge this is the first report of zen fish . Freshwat’s face , 5. Simi or soil . Steppin or soil . In thisM. senegalense that was originally found in Africa and has never been described elsewhere [M. houstonense have mostly been reported in the USA and Australia to date. This is the first study to report the existence of M. houstonense in China, widening the regional distribution of M. houstonense throughout the world.RGM are widely distributed and human infections due to RGM have been reported in most geographic areas of the world , with thlsewhere . AccordiMany experimental methods have been reported for drug susceptibility tests of RGM, such as agar disk elution, the E-test, broth microdilution and agar disk diffusion. Each method has its own advantages and disadvantages. The microbroth dilution method is recommended by the CLSI as a RGM drug sensitivity test, and is currently considered the gold standard. In the CLSI M24 A2 guidelines, the break points of the MIC method for amikacin, tobramycin, cefoxitin, clarithromycin, ciprofloxacin, moxifloxacin, doxycycline, imipenem, meropenem, linezolid and trimethoprim/sulfamethoxazole were specified . In thisM. houstonense infections. For the treatment of RGM, therapeutic regimens frequently vary depending on the nature of the disease. Single drug therapy for small or localized lesions is usually effective. However, in the case of diffuse or pulmonary infections, it is necessary to combine oral and injectable antibiotics [M. fortuitum group showed > 90% susceptibility or intermediate susceptibility to amikacin, cefoxitin, ciprofloxacin, gatifloxacin, imipenem, levofloxacin, linezolid, sulfamethoxazole or trimethoprim-sulfamethoxazole, and < 90% susceptibility or intermediate susceptibility to clarithromycin, doxycycline and vancomycin [M. houstonense isolate 14,133 appeared susceptible to clarithromycin (MIC < 4 μg/ml), but overnight incubation in clarithromycin (0.1 μg/ml) increased the clarithromycin MIC to > 128 μg/ml [erm genes are inducible at the molecular level. Therefore, it is necessary to be careful when interpreting the drug sensitivity of macrolides. M. houstonense is known to express the intrinsic macrolide erm gene, so inducible drug resistance may occur affecting the macrolide susceptibility results [M. houstonense was only susceptible to levofloxacin, moxifloxacin and amikacin. In this case, the patient was treated with a combination of levofloxacin and amikacin intravenous drip according to the drug sensitivity results. The therapeutic effect was good. However, a standardized treatment for M. houstonense remains to be proposed.Little is known about potential antimicrobial therapies to treat ibiotics . A reporncomycin . As prev results , 13. OurM. houstonense.A slight wound infection induced by RGM can heal itself and minor wound infections can be resolved be surgical debridement . Many caE. coli and M. houstonense. As the first report of infection caused by M. houstonense in China, it aids our understanding of the distribution of M. houstonense throughout the world.This case report describes an open fracture infection caused by combined infection of


Following publication of the original article , the autBH and JB conceptualised the study. JB supervised data collection, PH, AD and GGP analysed and interpreted the data and drafted the manuscript. BH, NW, and CS revised the manuscript critically. All authors read and approved the final manuscript.The (incorrect) text in this section says:PH drafted the manuscript. BH, NW, and CS revised the manuscript critically. All authors read and approved the final manuscript.BH and JB conceptualised the study. JB supervised data collection, PH, AD and GGP analysed and interpreted the data and As such, please be advised that the correct text (*correction in bold) is:


Cronobacter sakazakii was from a can of dried milk in 1950, although these organisms have likely existed for millions of years. In 1980, John J. Farmer III, proposed the name Enterobacter sakazakii for what had been known as “yellow-pigmented E. cloacae,” in honor of Japanese bacteriologist Riichi Sakazaki. Over the next decades, E. sakazakii was implicated in scores of cases of meningitis and sepsis among infants, frequently in association with powdered infant formula. In 2007, the genus Cronobacter was created to accommodate the biogroups of E. sakazakii, with C. sakazakii as the type species. The genus was named for Cronos, the Titan of Greek myth, who devoured his children as they were born (The first documented isolation of what would become known as ere born .


Trypanosoma that are transmitted cyclically by tsetse flies, mechanically by other haematophagous flies, or sexually. Trypanosoma congolense (subgenus Nannomonas) and T. vivax (subgenus Dutonella) are genetically and morphologically distinct from T. brucei, T. equiperdum and T. evansi (subgenus Trypanozoon). It remains controversial whether the three latter taxa should be considered distinct species. Recent outbreaks of surra and dourine in Europe illustrate the risk and consequences of importation of equine trypanosomosis with infected animals into non-endemic countries. Knowledge on the epidemiological situation is fragmentary since many endemic countries do not report the diseases to the World Organisation for Animal Health, OIE. Other major obstacles to the control of equine trypanosomosis are the lack of vaccines, the inability of drugs to cure the neurological stage of the disease, the inconsistent case definition and the limitations of current diagnostics. Especially in view of the ever-increasing movement of horses around the globe, there is not only the obvious need for reliable curative and prophylactic drugs but also for accurate diagnostic tests and algorithms. Unfortunately, clinical signs are not pathognomonic, parasitological tests are not sufficiently sensitive, serological tests miss sensitivity or specificity, and molecular tests cannot distinguish the taxa within the Trypanozoon subgenus. To address the limitations of the current diagnostics for equine trypanosomosis, we recommend studies into improved molecular and serological tests with the highest possible sensitivity and specificity. We realise that this is an ambitious goal, but it is dictated by needs at the point of care. However, depending on available treatment options, it may not always be necessary to identify which trypanosome taxon is responsible for a given infection.Equine trypanosomosis is a complex of infectious diseases called dourine, nagana and surra. It is caused by several species of the genus  Trypanosoma. They give rise to important economic losses in Africa, the Middle East, Asia and Latin America. Nevertheless, they can be considered as animal diseases that are seriously neglected, both by the scientific community as by veterinary authorities and regulatory organisations. The situation is aggravated by the reluctance of many endemic countries to notify dourine and surra to the World Organisation for Animal Health. Major obstacles to the local and global control of equine trypanosomosis are the lack of vaccines, the inability of drugs to cure the neurological stage of the infections, the inconsistent case definitions and the limitations of current diagnostics. Recent outbreaks of surra and dourine in Europe illustrate the risk and consequences of importation of equine trypanosomosis with infected animals into non-endemic countries. In view of the ever-increasing movement of horses around the globe, there is not only the obvious need for reliable curative and prophylactic drugs but also for accurate diagnostic tests and algorithms. The aim of this article is to provide an overview of the existing knowledge on the aetiology of equine trypanosomosis and the taxonomic status of the infecting trypanosomes, the geographical distribution, prevention and treatment options and diagnostic tools. We further discuss the issue of imperfect case definitions and propose research to address the limitations of current diagnostics.Equine trypanosomosis is a complex of infectious diseases called dourine, nagana and surra that are caused by several closely related species of Trypanosoma, including T. evansi, T. equiperdum, T. brucei, T. vivax, T. congolense and T. cruzi. Infections of horses with T. cruzi are very rare and are not further considered here [T. evansi), “dourine” (T. equiperdum) and “nagana”  but careful examination of published and unpublished data reveals that for all these three diseases, the clinical signs observed, including ventral oedema, emaciation, anaemia and neurological symptoms, can be very similar and are certainly not pathognomonic [Equine trypanosomosis is an infectious disease that is caused by several species of the genus red here . Historignomonic –7.Trypanosoma brucei and T. congolense are the only species that are confined to the distribution of tsetse flies (the vector) in sub-Saharan Africa. Trypanosoma equiperdum is transmitted sexually, and T. evansi is transmitted mechanically by blood-sucking flies, vampire bats, and possibly sexually [sexually , 9.T. evansi via contaminated meat or carcasses is well documented but normally does not occur in equines [Trypanosoma vivax can be transmitted both cyclically by tsetse flies and mechanically by other haematophagous flies. The global distribution of T. equiperdum, T. evansi and T. vivax is much wider, including Africa and Latin America for T. vivax, Africa, Latin America and Asia with sporadic import cases in Europe for T. evansi and worldwide, except Oceania, for T. equiperdum [T. brucei, T. congolense, T. equiperdum and T. vivax in horses. However, evidence is increasingly being published on horses infected with T. evansi, T. equiperdum and T. vivax in Brazil, Ethiopia, India, Israel, Jordan, Mongolia, Nigeria, Pakistan, South East Asia, Sudan, Venezuela, etc. [Oral transmission of  equines , 11. Tryuiperdum , 13. Mosuiperdum . From reuiperdum –15. No sla, etc. –25.T. congolense, all the other trypanosomes are known to reside mainly in extravascular spaces of many tissues and organs, including the central nervous system. Evidence is accumulating that none of the aforementioned drugs is effective in the neurological stage of the disease since none is able to cross the blood-brain-barrier in sufficient amounts [T. evansi) in Spain was brought under control upon treatment with melarsomine hydrochloride  of all parasitologically confirmed and suspect animals  as well as of all animals that were in direct or indirect contact with the index case [T. equiperdum), occurred in Italy in 2011 and illustrates the risk of importation of equine trypanosomosis with infected animals into non-endemic countries [Vaccines against equine trypanosomosis do not exist. Chemotherapy of equine trypanosomosis consists of treatment with diminazene diaceturate, isometamidium chloride, quinapyramine chloride/quinapyramine sulphate combination, suramin or melarsomine hydrochloride. Except for trypanosome strains that display an innate or acquired resistance, these drugs are able to clear the parasites from the blood circulation . However amounts , 27–30. dex case . The latdex case , 32. Anoountries . That ouountries , 33, 34.Trypanosoma congolense (subgenus Nannomonas) and T. vivax (subgenus Duttonella) are species that are clearly separated, both genetically and morphologically, from the other taxa within the genus Trypanosoma. On the other hand, whether T. brucei, T. equiperdum and T. evansi, traditionally grouped together under the subgenus Trypanozoon, can be considered distinct species remains controversial. Morphologically, T. brucei can be distinguished from the latter two taxa by its pleomorphic nature with long slender, intermediate and short stumpy trypomastigote forms present in the mammalian host. The short stumpy forms are those that will initiate infection in the vector of nagana, the tsetse fly [Trypanosoma equiperdum and T. evansi are monomorphic and display only long slender forms, although in T. evansi, intermediate forms may sometimes be found [T. equiperdum and T. evansi, and the fact that genetic differences to T. brucei are small, have led to proposals that they should be classified as subspecies or strains of T. brucei [T. equiperdum isolates are in fact misidentified T. evansi strains, and that T. evansi and T. equiperdum evolved separately and on more than one occasion from T. brucei [T. evansi is further divided into T. evansi type A and type B, the latter so far only isolated from camels in eastern Africa [T. equiperdum strains can be divided in at least two and maybe more clades of distinct evolutionary origin [T. equiperdum strains from outbreaks in Italy and in Mongolia may even further complicate the taxonomic situation [Trypanosoma taxa that may cause trypanosomosis in equines.etse fly . Trypanobe found . Lack of. brucei –38. Othe. brucei . Today, n Africa . The “try origin . Newly iituation , 16. TabDiagnosis of equine trypanosomosis can be challenging due to the absence of specific clinical signs, and because parasitaemias in infected hosts are usually below the detection limit of parasitological tests and can even be below the detection limit of molecular DNA tests . TherefoClinical signs of nagana, dourine and surra vary with disease stage , infecting strain and variable intra- and inter-species susceptibility of the host.T. brucei, T. equiperdum or T. evansi, ataxia and paralysis of the hind quarter and lips usually precede death. The clinical course of the disease and long-term outcome depend on the parasite strain involved and the immunological status of the host. Other equine diseases that share some clinical signs with trypanosomosis are equine viral arteritis, equine infectious anaemia, contagious equine metritis, anthrax [For example, donkeys can carry an infection without developing classical signs of the diseases . Typical anthrax .T. vivax and T. congolense from the Trypanozoon species based on morphology, but this is not evident for the species within the subgenus Trypanozoon. In case of a microscopic trypanosome-positive specimen, species identification is best performed with a combination of molecular tests that have also the advantage of having a lower detection limit than microscopic parasite detection and that are able to detect mixed infections that may remain cryptic by microscopic examination [For microscopic parasitological diagnosis, the most common biological fluid examined is blood, but parasites may be detected in lymph aspirated from superficial lymph nodes, cerebrospinal fluid, milk and vaginal or preputial discharges , 49, 50.mination .18S and ITS1 sequences within the ribosomal DNA (rDNA) locus as well as satellite DNA are preferential targets for species-specific molecular diagnostics because of their multi-copy nature and the possibility to discriminate the taxa of the subgenus Trypanozoon from T. congolense and T. vivax [Trypanozoon is more challenging. PCR tests based on the mitochondrial DNA of these parasites (kinetoplast DNA or kDNA) can potentially distinguish T. evansi and some T. equiperdum from T. brucei by the absence of the maxicircle component of kDNA, and identify T. evansi type B by presence of the type B minicircle [T. evansi strains are naturally akinetoplastic (i.e. complete lack of kDNA) so that molecular tests targeting kDNA have limited value. Some authors claim that T. evansi type A can be identified by the presence of the type A-specific RoTat 1.2 variant surface glycoprotein (VSG) gene and T. evansi type B by the presence of the type B-specific JN 2118HU VSG [T. brucei has not been investigated extensively. Furthermore, the VSG repertoire is subject to recombination and therefore inherently unstable, and other researchers have shown that some Kenyan Trypanozoon isolates with type A minicircles appear to lack the RoTat 1.2 gene, while the JN 2118Hu VSG gene was also found in T. b. gambiense type II [T. brucei from T. equiperdum, which means that the epidemiological context has to be taken into account to identify the trypanosome species in an infected horse.A plethora of molecular diagnostic tests has been described which target DNA sequences that are specific at different taxonomic levels and which use diverse technology. The most commonly used test formats are polymerase chain reaction (PCR) and quantitative PCR (qPCR). The T. vivax , 51–54. nicircle , 55, 56.18HU VSG –59. Howe type II . UnfortuT. congolense and, to a lesser extent, T. vivax, Trypanozoon trypanosomes are primarily tissue parasites; parasitaemia is often very low and seldom reaches the threshold of current parasitological or even molecular diagnostic tests, especially in asymptomatic carriers. Therefore, antibody detection tests are available and can provide indirect evidence of infection.In contrast to T. equiperdum infection [T. evansi infection, OIE recommends IFAT, an enzyme linked immunosorbent assay (ELISA/RoTat 1.2), a card agglutination test for trypanosomosis using T. evansi antigen (CATT/T. evansi) and immune trypanolysis (IT) [T. brucei, T. vivax and T. congolense, OIE recommends using IFAT and ELISA [The complement fixation test (CFT) and the indirect fluorescence antibody test (IFAT) are the only OIE recommended tests for nfection . For T. sis (IT) . For infnd ELISA .T. evansi type A [T. evansi test is a direct agglutination test that makes use of the same T. evansi RoTat 1.2 clone used in the IT test, but the preparation of the reagent exposes other surface antigens as well, resulting in some cross-reactivity with T. vivax and T. equiperdum [The IT test detects exclusively antibodies that recognise one single multi-copy epitope at the surface of the RoTat 1.2 VSG expressing i type A . As a reuiperdum , 65, 66.in vivo (mice or rats) or propagated in vitro [T. equiperdum strain, has the advantage that it, to a certain level, also reacts with antibodies against T. evansi; however, this also entails the risk of false positive reactions [T. equiperdum infection is considered non-curable, any false positive reaction will have serious consequences . Reactivity of CFT with horses infected with T. brucei, or even with T. congolense or T. vivax, seems possible but remains undocumented. An additional problem with CFT is that reactivity may vary in function of the trypanosome strain used to prepare the antigen and of the VSG expressed by the trypanosomes at the moment of harvesting. Efforts to harmonise the antigen preparation of CFT in the diverse laboratories around the world are challenged by the fact that, within the subgenus Trypanozoon, genetic distinction between T. equiperdum, T. evansi and T. brucei is blurred, as discussed above [T. equiperdum for the purpose of CFT antigen preparation. It is this strain that is now used by the European Reference Laboratory for Equine Diseases in France, the National Reference Laboratory for Dourine in Germany and the USDA National Veterinary Services Laboratories [T. equiperdum isolates have not been fully typed; whether they may serve as reference strains for the Asian region remains to be investigated [Other serological tests use crude preparations derived from bloodstream form trypomastigotes grown in vitro , 68. Theeactions . Since Ted above , 69, 70.ed above , the conratories , 68. Newstigated .T. evansi (other than RoTat 1.2), T. brucei, T. vivax and T. congolense have not been defined. Furthermore, as a result of antigenic variation, it is not possible to control the particular VSG that is expressed at the moment of antigen preparation. Since this VSG accounts for 10% of the protein content of a bloodstream form trypomastigote, it is a major, yet undefined, component of crude antigen preparations for IFAT and ELISA. Other proteins in crude trypanosome antigen preparations may cross-react with antibodies unrelated to infection with trypanosomes [Similar to the CFT, the trypanosome strains used for IFAT and ELISA for anosomes .Trypanozoon parasites that can be reliably distinguished from T. brucei and T. evansi and (ii) if CFT, IFAT and PCR exist that are fully specific for these parasites. Unfortunately, both conditions are not fulfilled.Case definitions with unequivocal criteria to identify a case of dourine, surra and nagana are essential to estimate the impact of these diseases on the horse population, to guide surveillance, control and prevention strategies, and to choose among treatment options, the latter taking into account that treatment success is largely dependent on the stage of the diseases rather than on the infecting trypanosome strain. An attempt at such case definition was made during the dourine outbreak in Italy in 2011; subsequently, the same case definition was adopted by the OIE Terrestrial Manual , 33. AccT. evansi (anti-horse IgG whole molecule), CATT/T. evansi, PCR-TBR (i.e. PCR targeting Trypanozoon-specific satellite DNA [Trypanozoon spp. (sic) if it is positive to PCR-TBR and/or if Trypanozoon parasites are observed by microscopic examination. An equid is considered as seropositive to surra if it is positive to ELISA-T. evansi and/or CATT/T. evansi; in this case, the animal should be tested for CFT-dourine, and, if it is positive for CFT-dourine, it is also considered as seropositive to dourine; if it is negative to CFT-dourine, it is considered as seropositive to surra only [Trypanozoon taxa, but the definition of seropositivity is questionable and does not take into account possible cross-reactions with T. brucei, T. vivax and T. congolense. It also disregards the possibility to use more specific tests such as the IT and the ELISA/RoTat 1.2 to rule out or suggest infection with T. evansi type A. Both may miss T. evansi type B infections, but T. evansi type B has so far only been isolated from dromedary camels in Kenya and Ethiopia [According to the OIE Terrestrial Manual , an equilite DNA ) and micrra only . This “cEthiopia , 56.T. brucei, T. vivax and T. congolense, the OIE Terrestrial Manual does not provide any case definition.For equine infections with In order to improve the specificity of diagnostics for equine trypanosomosis, the focus should be on molecular and serological tests.T. congolense, T. vivax and Trypanozoon taxa. However, within Trypanozoon, no single test is able to identify each taxon unequivocally. On the other hand, for the purpose of international movement of horses, it seems sufficient to use a combination of genus- and subgenus-specific molecular tests. As qPCR is becoming the standard now, the conventional PCR tests might be transformed into qPCR to improve standardisation and sensitivity. In any case, given the usually low parasitaemia in clinically healthy, yet infected, equines, the negative predictive value of molecular tests is inherently low.Molecular tests can identify with relatively high sensitivity and excellent specificity T. congolense and T. vivax, recombinant fragments of respectively Cathepsin B1 and the cytoskeleton associated protein GM6 are used in the VeryDiag test [T. vivax Cathepsin L has some diagnostic potential [Trypanozoon, many common proteins have already been expressed as recombinant antigens with proven diagnostic potential, in particular the invariant surface glycoproteins 65 and 75 , the cytoskeleton associated protein GM6 and, more recently, a T. equiperdum protein that exhibits homology with the regulatory subunit of mammalian cAMP-dependent protein kinases [Trypanozoon infections in general, irrespective of the infecting trypanosome species and geographical origin. In case there is a need to detect T. evansi type A specific antibodies, a recombinant fragment of VSG RoTat 1.2, expressed in different systems, has shown its diagnostic potential [Regarding serological tests, improvements are possible with respect to test format and antigens. CFT and IFAT should be replaced by techniques that are less complex and easier to standardise, such as ELISA for high throughput or immunochromatography for individual testing. Important to keep in mind, however, is the market failure that prevents commercial companies to invest in the development of such tests. The diagnostic specificity of antibody detection tests is largely defined by the antigen preparation. Therefore, the use of crude antigen preparations should be discouraged, and initiatives to replace them with recombinant or synthetic peptides should be supported. For iag test . Also, rotential . For the kinases –86. Noneotential , 87–89.Trypanozoon. We propose to conduct studies into (i) improved molecular tests with the highest possible sensitivity; and (ii) improved serological tests with the highest possible specificity. For the proper evaluation of these diagnostics, it will be necessary to establish a panel of representative trypanosome strains, and their corresponding DNA/RNA, as well as a panel of biological specimens (serum/plasma/blood) from horses naturally or experimentally infected with T. vivax, T. congolense and representative Trypanozoon taxa.To address the limitations of the current diagnostics for equine trypanosomosis we recommend not to aim for the distinction between taxa within the subgenus