Sillago species, the black-banded sillago, Sillagonigrofasciatasp. nov., is described based on 302 specimens sampled from the southern coast of China. Morphological comparisons have been conducted between the new species and ten other Sillago species. The results show that the new species is characterized by a black mid-lateral band below the lateral line when fresh; other characteristics are similar to those of Sillagosihama but subtle differences exist on the swim bladder between Sillagonigrofasciata sp. nov. and S.sihama. A detailed description and illustrations are provided for the new species. The validity of this new species is also supported by a genetic comparison using sequences of the mitochondrial cytochrome c oxidase subunit I (COI) gene.A new  Sillaginidae Richardson, 1846, commonly known as sand whiting or sand borer, is a small family of demersal marine fishes that primarily inhabit inshore waters with sandy substrates or estuarine areas of rivers throughout the Indo-West Pacific  exhibits many cryptic lineages across its Indo-West Pacific distribution ; Sillago Cuvier, 1817 ; and Parasillago McKay, 1985 (swim bladder with a single posterior extension and the duct-like process). subgenusSillago . The presence of two posterior extensions of the swim bladder observed in five species suggested that they should belong to the subgenusSillago. In addition, a redescription of S.indica McKay, Dutt & Sujatha, 1985, reassigns it to the subgenusSillago  and Fishery Ecology Laboratory, Fisheries College, Ocean University of China, Qingdao (OUC_FEL).The unidentified specimens were collected from the southern coast of China, more precisely in Fuding , Xiamen , Changhua , Chiayi , Shantou , Zhuhai , Zhanjiang , Beihai , Fangchenggang , Haikou , and Danzhou   DNA cytochrome oxidase subunit I (COI) fragments of Sillago spp. were amplified based on the method of PCR) followed the protocols of NJ) tree under the Kimura 2-parameter (K2P) model. COI sequences of S.nigrofasciata sp. nov. obtained in the present study were submitted to GenBank with the following accession numbers: KU051808, KU051809, KU051812, MG571453–MG571458, MG911029–MG911030. Twenty-nine COI sequences were obtained from GenBank with the following accession numbers: S.indica (KM250229–KM250232), S.panhwari , S.parvisquamis (HQ389247–HQ389249), S.shaoi , S.sihama , S.sinica , and S.suezensis (FJ155362–FJ155364). Sillaginodespunctata was selected as the outgroup for genetic analyses based on a previous phylogenetic hypothesis of To analyze genetic differences between this new species and other congeners, mitochondrial  were shown in Table NJ tree based on the COI gene sequences revealed that all previously recognized and the newly discovered S.nigrofasciata sp. nov. individuals formed monophyletic groups , I+20–22, soft anal fin rays 20–22; scales in lateral line 67–75, scales above lateral line 4–6; gill rakers 2–4+5–8; vertebra: abdominal 14 or 15 (mostly 14), modified 3–7 (mostly 4 or 5), caudal 13–18, and total 34 or 35 (mostly 34)  in SL. Snout long, 41.7% (39.6–52.7%) of HL. Eye moderate, its margin slightly covered with adipose eyelid, diameter 17.9% (16.5–24.9%) of HL. Interorbital region flat, interorbital width 23.5% (14.8–31.2%) of HL. Nostrils situated anterior to upper margin of eye; posterior margin of anterior nostril with single anteriorly directed flap; posterior nostril lacking flap. Mouth small, terminal, anterior tip of upper jaw situated at almost same position as tip of lower jaw. Upper jaw with small canines forming a wide tooth band becoming narrower posteriorly. Lower jaw with small canines, forming tooth band anteriorly, width same as upper jaw tooth band, tooth band gradually becoming narrower posteriorly, ending in one row. Palatine and tongue toothless. Vomer with three to four rows of canine teeth. Posterior margin of preopercle slightly serrated. Gill aperture large, lateral, extending to ventral side of head, stopping at middle bottom of opercle. Gill rakers on the first arch pointed but short. Caudal peduncle short, depth of caudal peduncle 61.0% (51.3–88.5%) of length of caudal peduncle.General body features are shown in Figure Body covered with small or moderate sized ctenoid scales, and cheek scales cycloid, arranged in two or three rows. Lower part of pre-opercular-mandibular canal covered with cycloid scales. The base of pectoral fin and ventral fin lacking scales. Lateral line beginning above gill aperture and anterior portion of pectoral fin, extending along curve of dorsal edge to the end of body.Two separated dorsal fins, first dorsal fin XI (X–XII), obviously higher than second, origin posterior to top of pectoral fin base, composed of spines, gradually shortening. Fin membrane with dense black spots. Base of second dorsal fin long, composed of a single spine and 21 (20–22) soft rays, originating mid-body, and not extending to caudal fin origin when placed flat. Origins of anal fin slightly posterior to cloacal pore, with II+22 (20–22), not extending to caudal fin origin when placed flat. Pectoral fin 16 (14–16), slender. Two separated ventral fins broad, I+5, approximately triangular, and shorter than pectoral fin.Upper surface of head dark brown and trunk bright brown, grading to silver on abdomen. Dorsal side of snout brownish gray. Cheek yellow, slightly silver posteriorly, with black dots amassed on the anterior inferior part of eyes. A wide faint stripe composed of tiny black dots on skin always present, from opercular to caudal peduncle. Dorsal fins yellowish hyaline, small dark dense spots on fin membrane. Pectoral, ventral, and anal fins yellowish hyaline with dark spots; caudal fin yellowish dusky with a black margin and grayish brown margin posteriorly, lobes usually broadly truncated posteriorly.Swim bladder large. Two anterior extensions diverging to terminate on either side of the basioccipital above the auditory capsule. Two posterior tapering extensions of the swim bladder penetrating into the caudal region, one usually longer than the other. Two anterolateral extensions originate anteriorly, each branch into anterior and posterior sub-extensions: the anterior one comprising a short, simple blind tubule and the posterior sub-extensions kinked, long and complex, extending along the abdominal wall ventral to the base of the posterior extensions, respectively, tangent but not interconnected. A single duct-like process originating from ventral surface of swim bladder extending to the urogenital opening and a sub-extension connecting with a sanguineous vesicle close to vertebra, of unknown function. Eight or nine lateral processes extending from entire lateral surface of main body of swim bladder, anterior three or four stout and horn-like, posterior five or six small and triangular in shape.S.sihama in nearshore areas and frequently entering estuaries for considerable periods, it is common along the beaches, sand bars, and mangrove creeks with sandy substrates. Depths ranging from 0 to 20 m, and frequently captured by trawling vessels.Habitat is similar to Sillagonigrofasciata sp. nov. was only found along the southern coast of China including the coastal waters of the South China Sea and the Taiwan Strait. Actually, its distribution range is similar to that of S.sihama in China , from S.parvisquamis and S.sinica by the dusky spots on the second dorsal fin membranes (five or six rows in S.parvisquamis and three or four rows in S.sinica). Empirically, S.nigrofasciata sp. nov. could also be distinguished from S.sihama, S.indica, S.panhwari, and S.suezensis by the coloration of anal fin .According to the subgeneric grading system in S.nigrofasciata sp. nov. was easily distinguishable from other species by the following: S.megacephalus by having a smaller head  and less soft rays in anal fin , from S.parvisquamis and S.sinica by having 34–35 total vertebrae (39–40 in S.parvisquamis and 37–39 in S.sinica), from S.parvisquamis and S.sinica can also by having 67–75 scales on lateral line (79–84 in S.parvisquamis and 75–79 in S.sinica), and from S.caudicula by gill rakers (4/11 in S.caudicula) and soft rays in anal fin (23–24 in S.caudicula).Moreover, by the primary diagnostic features Table , S.nigrS.suezensis was always controversial (S.suezensis (Mediterranean population) and S.sihama (Hong Kong and southern Red Sea populations) showed a strong genetic divergence . Sillagonigrofasciata sp. nov. could also be easily distinguished from S.intermedius and S.caudicula by those swim bladders with simple anterolateral extensions; S.parvisquamis stood out as having the strongest anterolateral extensions in comparison with the others , which indicated that the COI gene used as a barcode is an effective tool to identify Sillago species.Species-level taxonomy for  bladder . Differe bladder . This stes level . This stSillago species, this species could be confused with S.sihama based on the countable characters and the shape of the swim bladder. In fact, as one clade of S.sihama complex  and the swim bladder (the origin of the duct-like process of S.nigrofasciata sp. nov. is at the terminus of the swim bladder and starting at the joint of the roots of two posterior extensions). Sillagosihama lacks the mid-lateral stripe and dark spots on anal fin, and its origin of the duct-like process is anterior to the terminus of the swim bladder and anterior to the joint of the roots of both posterior extensions.According to the conventional classification of der Fig. , but witder Fig. . S.nigrSillagonigrofasciata sp. nov. overlaps with that of S.sihama in China: the coastal waters of the South China Sea and the Taiwan Strait. Actually, S.sihama across its Indo-West Pacific range exhibits by far the highest levels of genetic diversity. A few new species have been described in this range across the IWP: S.caudicula (from Oman and Madagascar), S.sinica (from China), S.suezensis (from the northern Red Sea and Mediterranean), S.shaoi (from Taiwan Strait), and S.panhwari (from the northern Arabian Sea), and they have been always regarded as junior synonyms of S.sihama (S.sihama range that probably represent species-level taxa. These findings indicate a thorough taxonomic review of S.sihama (and its junior synonyms) is necessary (At present, the distribution of .sihama . Howeverecessary .


Candidiasis can be present as a cutaneous, mucosal or deep-seated organ infection, which is caused by more than 20 types of Candida sp., with C. albicans being the most common. These are pathogenic yeast and are usually present in the normal microbiome. High-risk individuals are patients of human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS), organ transplant, and diabetes. During infection, pathogens can adhere to complement receptors and various extracellular matrix proteins in the oral and vaginal cavity. Oral and vaginal Candidiasis results from the overgrowth of Candida sp. in the hosts, causing penetration of the oral and vaginal tissues. Symptoms include white patches in the mouth, tongue, throat, and itchiness or burning of genitalia. Diagnosis involves visual examination, microscopic analysis, or culturing. These infections are treated with a variety of antifungals that target different biosynthetic pathways of the pathogen. For example, echinochandins target cell wall biosynthesis, while allylamines, azoles, and morpholines target ergosterol biosynthesis, and 5-Flucytosine (5FC) targets nucleic acid biosynthesis. Azoles are commonly used in therapeutics, however, because of its fungistatic nature, Candida sp. evolve azole resistance. Besides azoles, Candida sp. also acquire resistance to polyenes, echinochandins, and 5FC. This review discusses, in detail, the drug resistance mechanisms adapted by Candida sp. Candida species are commensals and thus are part of the normal human flora and are localized on skin and gastrointestinal and genital tracts. However, Candida can also cause various infections in susceptible patients that includes elderly, hospitalized, or immunosuppressed patients. Invasive Candida infection is one of the most common fungal infections globally [Candida sp. were reported to be one of the leading causes of healthcare-associated infections [Candida sp., Candida albicans is the most commonly recovered (37%) from clinical species, followed by Candida glabrata (27%). Other clinically relevant species recovered from blood stream infections include C. parapsilosis (14%), C. krusei (2%), C. tropicalis (8%), C. dubliniensis (2%), C. lusitaniae (2%), and the most recent, C. auris. C. auris is an emergent multi-drug-resistant pathogen that is often misidentified and at present is a major concern in healthcare settings. C. auris reported cases increased by 318% between 2015 and 2018. As per the Centers for Disease Control and Prevention (CDC), an estimated 34,000 cases of Candidiasis were reported in hospitalized patients and about 1700 people died in 2017 [Candidiasis has a diverse clinical spectrum ranging from non-life threatening superficial mucocutaneous infections to devastating invasive disease associated with candidemia. In fact, the attributable mortality observed with candidemia is between 30% and 47% [Candida infection is also commonly associated with medical devices such as central venous catheters, cardiovascular devices, and urinary catheters [globally . In the fections . Amongstglabrata 7%. Otherecovered 7% from csilosis 1%, C. kruCandidiasis, most Candidiasis cases are still diagnosed by routine fungal cultures of blood, urine tissue, and other body fluids. This method can have a low sensitivity, and in some cases, it can also render false-positive results because of contamination, which can occur in the process. Commonly, empiric anti-fungal therapy is initiated in febrile or septic patients in the intensive care unit with an indwelling central venous catheter, recent abdominal surgery, or chemotherapy in the absence of response to anti-microbial therapy. This approach can lead to the unnecessary use of antifungal agents and promote the emergence of resistance in individuals without invasive Candidiasis or a delay in effective antifungal therapy for those who are infected.Because of a lack of rapid diagnostic assays for invasive Candidiasis depends on the patient’s immune status, location, and severity of the infection. In addition to an adequate source control, removal of infected medical devices and antifungal agents have been important therapeutic tools for invasive Candida infections [Candida species are available, including polyenes, azoles, echinocandins, and 5-Flucytosine (5FC). Similar to antibiotics for bacterial infection, an emergence of antifungal resistance among Candida species is a serious threat to public health worldwide. According to the US Center of Disease Control and Prevention (CDC) 2019 report of antibiotic resistance threat, more than 34,000 cases and 1700 deaths annually were due to drug-resistant Candida sp. In addition, 323 cases of emerging multidrug- resistant Candida auris infection were reported.The strategy for treatment of invasive fections . At presCandida sp.In the current review, we discuss the molecular mechanisms of action of these antifungal agents as well as mechanisms of drug resistance used by Candidiasis can be treated with antifungals that belong to different classes of drugs and target different cellular processes, thereby either inhibiting (fungistatic) or killing  the growth of this pathogenic yeast. These cellular processes include the biosynthesis of the cell wall, cell membrane, and biosynthesis of RNA. Each of these biosynthesis processes involves a series of enzymes. The targets and the mechanisms of antifungals employed for treating Candidiasis are outlined below.Ergosterol is the major sterol component of fungal cell membranes, including the plasma and mitochondrial membranes. It is vital for fungi to maintain the structure and function of these membranes. Together, sterols and sphingolipids form lipid rafts in the cell membrane. These lipid rafts contain many biologically important proteins, which are involved in signaling, response to stress, mating, and nutrient transport ,7. ErgosAzoles: Azoles are five-membered heterocyclic compounds with antifungal properties. They are broadly classified into two groups that include imidazole and triazole. Imidazoles contain two nitrogen in the azole ring and include antifungals clotrimazole, econazole, ketoconazole, miconazole, and tioconazole. Triazoles contain three nitrogen in the azole ring and include fluconazole, itraconazole, voriconazole, isavuconazole, and posaconazole. Fluconazole is the most common azole used during therapy. Second generation of triazoles that include voriconazole, posaconazole, and isavuconazole are more potent against resistant pathogens. Isavuconazole is a novel azole, as effective as voriconazole but less toxic than voriconazole. Toxicity is lower in isavuconazole, since it is highly water soluble and does not require beta-cyclodextrin in its intravenous (IV) formulation, which is required for voriconazole, causing reduced nephrotoxicity [toxicity ,9. Isavutoxicity ,11.Candida infections. Azoles target the enzyme 14α–demethylase (Erg11p), an important enzyme in ergosterol biosynthesis  in the ergosterol biosynthesis pathways [pathways . These dpathways .Morpholines: This class of drugs include fenpropimorph, tridemorph, and amorpholine. They target the ergosterol biosynthetic enzyme C-14 sterol reductase (Erg24p) [(Erg24p) . The mor(Erg24p) . Only 5%(Erg24p) .Candida sp. is also targeted by various antifungals. The cell wall is the rigid outermost layer of the fungal cell and is the first line of defense, protecting the fungal cells from osmotic stress. Mammalian cells do not have cell walls and therefore the enzymes of the cell wall biosynthetic pathways are important antifungal targets [FKS1, FKS2, and FKS3 [Besides ergosterol, cell wall biosynthesis of  targets . Echinocand FKS3 . β1-3 gland FKS3 . These dand FKS3 . Echinocand FKS3 .Other important antifungals are those that target nucleic acid biosynthesis. These were originally designed in the 1950s and are one of the oldest classes of antifungal drugs. 5-Flucytosine (5FC) is an antifungal that interferes with nucleic acid biosynthesis. The susceptible cells import 5FC via the cytosine permease enzyme . 5FC is Several important factors may contribute to treatment failure. These are listed below.Bioavailability: Differences in drug bioavailability between different tissues can contribute to the effectiveness of antifungal agents in treating fungal infections. For example, bioavailability of azole drugs in vaginal tissues with a low pH is much lower than that in blood [Candida infections in the urinary tract. Importantly, different lipid formulations vary in their tissue distribution and toxicity. Specifically, although LAMB has a better overall therapeutic index, ABLC achieves higher lung and kidney tissue concentrations [in blood . Also, itrations .Host Immune system: The host immune system plays a critical role in successful therapeutics. Especially, static antifungal drugs such as the azoles require support from the host’s immune system for successful treatment of invasive Candida infections. In severely neutropenic patients, these drugs may be less effective and supportive treatment with growth factors to boost engraftment is required.Candida sp. can colonize the mouth of 64% to 84% of human immunodeficiency virus HIV-infected patients [+ cell counts (<50/mm3) [Candida sp. In one case, a series of 17 C. albicans clinical isolates were obtained from a single HIV patient receiving azole therapy over two years. Azole-resistance increased steadily as these isolates were exposed to increasing amounts of drugs [Successful azole therapy depends on the drug and on the host’s immune response to the fungus. Without a functional immune system, azoles cannot control the fungal infections. For example, patients . Develop<50/mm3) . These pof drugs .Biofilm formation: Fungi can develop biofilms on the surface of medical equipment, such as catheters. These biofilms are not only resistant to the penetration of some antifungals but also the drug targets may be less expressed [xpressed .Drug pharmacokinetics, infection severity and side-effects: The pharmacokinetics of the drugs is an important factor that contribute to drug efficacies. These include drug metabolism, absorption, and distribution that can alter drug effectiveness. Additionally, drug efficacy can depend on the severity of the infection, and the population size of the infecting organisms [Candida sp. [rganisms . Furtherdida sp. . Toxicitdida sp. . Nephrotdida sp. ,31. Amphdida sp. . Azoles dida sp. . Similardida sp. .Antifungal drug resistance: Another important factor that can cause therapeutic failure is antifungal drug resistance of the infecting Candida sp. For example, prolonged use of a fungistatic azole drugs like fluconazole may cause the pathogenic yeasts to develop resistance, making the drug less effective. Fluconazole is the most common azole used for prophylaxis and treatment of Candida infections. However, several Candida sp. have evolved resistance to azoles, which is an emerging problem causing therapeutic failures [C. albicans became a severe problem in the 1990s, when 90% of acquired immunodeficiency syndrome (AIDS) patients had oral candidiasis, many received a long-term azole therapy and certain patient populations developed resistance [failures . For exasistance .For decades, extensive research has analyzed the molecular mechanisms of azole resistance in pathogenic fungi. The known mechanisms are described below and summarized in Candida clinical isolates.The different mechanisms of azole resistance are described below. These mechanisms are all well studied, are all equally important, and are frequently observed in several drug-resistant C. albicans [A large number of membrane proteins are present in pathogenic yeast. These membrane proteins are present in the cell membrane , vacuolaalbicans .a) ABC-transporters. Adenosine Triphosphate (ATP)-binding cassette transporters (ABC-T) are active transporters requiring ATP as an energy source. Each ABC-T consists of two membrane-spanning domains (MSD), each containing six transmembrane segments, and two nucleotide binding domains (NBD). Each NBD consists of an ATP-binding cassette (ABC) that binds ATP [inds ATP . Azoles inds ATP .b) MFS-transporters. Major facilitator transporters (MFS-T) require a proton gradient of the plasma membrane as an energy source to transport xenobiotics. MFS-Ts do not have the NBDs that are characteristic of ABC-Ts, and they have 12 to 14 transmembrane segments [segments . Azoles Two types of membrane transporters have been identified that correlate to azole resistance in fungi. They are described below  and CaPDR16 (an ABC-T) has been observed in azole-resistant isolates [C. albicans genome contains many other membrane transporters. However, to date, none have been shown to effect azole resistance [Besides isolates . CaPdr16isolates . CaFlu1pisolates . The C. sistance .CaCDR1 and CaCDR2 are regulated by transcription factor CaTac1p, while CaMDR1 is regulated by transcription factor CaMrr1p [2-Cys6). Gain of function (GOF) mutations in CaTAC1 and in CaMRR1 result in constitutive overexpression of their respective pumps, and azole resistance in many clinical isolates Δ [The efflux pumps  CaMrr1p ,49. Bothisolates . GOF mutisolates ,52. GOFs7-I998)Δ .CaCDR1 and CaCDR2, including the transcription factors, CaNDT80, CaFCR1, and CaFCR3 [CaNDT80, CaFCR1, and CaFCR3 with resistance in clinical isolates. CaMDR1 is transcriptionally regulated by CaMRR1 as well as CaCAP1 and CaMCM1 [There are other transcription factors that regulate the expression of d CaFCR3 ,55. To dd CaMCM1 . In vitrd CaMCM1 .C. glabrata, overexpression of the ABC-T efflux pumps CgCDR1, CgSNQ2, and CgPDH1 has been shown to contribute to azole resistance in clinical isolates [CgPDR16 has also been associated with the azole resistance mechanism in C. glabrata [CgFLR1 and CgQDR2 is also observed in azole-resistant clinical isolates [In isolates ,59. Overglabrata ,61. Amonisolates ,63.CgCDR1, CgSNQ2, and CgPDH1 [CgPDR1 also regulate virulence. For example, GOF L280F in PDR1 regulates adherence to macrophages and macrophage-mediated phagocytosis [CgPDR1 results in increased expression of adhesin proteins like Epa1, causing increased adherence [CgPDR1 also regulates MFS-T CgQDR2, which upregulates in response to azoles [CgQDR2 is caused by GOF L946S in CgPDR1. Deletion of CgQDR2 showed susceptibility to azole, miconazole, suggesting the role of CgQDR2 in azole resistance [GOF mutations in the transcription factor CgPdr1p activates CgPdr1p, which in turn induces the ABC-T pumps d CgPDH1 . These Gd CgPDH1 . GOF mutocytosis . GOF in dherence . CgPDR1 o azoles . This upsistance .C. glabrata. Loss of mitochondria leads to azole resistance, which correlated with upregulation of CgCDR1, CgPDH1, and other genes [Mitochondria also plays an important role in efflux pump-mediated azole resistance in er genes .C. krusei. In clinical isolates of C. krusei, overexpression of the ABC-T pumps CkAbc1p and CkAbc2p correlates with drug resistance, expression of both pumps can be induced by azoles, and expression of the pumps in a hyper-susceptible S. cerevisiae strain results in resistance [C. krusei [C. parapsilosis and C. dubliniensis [Membrane transporter overexpression can also affect intrinsically azole-resistant sistance . However. krusei . Increasiniensis ,71.C. auris, an emerging multidrug-resistant fungal pathogen [C. auris contains C. albicans homolog CDR1, which was found to be overexpressed in generationally aged C. auris that showed increased tolerance to fluconazole [C. albicans homolog of CDR1 contributed in the evolution of azole-resistant C. auris isolates [TAC1 in the evolved strain when compared to its parent [Membrane transporter-based azole resistance was also recently observed in pathogen ,73. C. aconazole . Recentlisolates . This ins parent .Mutation and/or overexpression of ergosterol pathway genes - trienol during ergosterol biosynthesis  in UPC2  and aneuploidy can correlate with azole resistance in fungi. If one allele of a gene is mutated, LOH can copy the mutation to the second allele. In clinical isolates of sistance . Clinicasistance . Additiosistance . Howeversistance . Furthersistance ,106. Triand CDR2 , and incpression . Aneuplopression . Chromosexposure .C. glabrata. For example, segmental rearrangements were previously observed in chromosome M and F in azole-resistant C. glabrata clinical isolates. Chromosome M harbors efflux pump-encoding gene CgCDR1, which was duplicated in these isolates, while chromosome F harbors efflux pump-encoding gene CgPDH1 [ERG11 in C. glabrata caused increased azole resistance [CDR1 and ERG11 was also observed in generationally aging C. auris cells that rendered increased resilience to fluconazole [Chromosomal alterations are also observed in e CgPDH1 . Besidessistance . Recentlconazole .C. albicans [Several studies have hypothesized that defective azole import may contribute to drug resistance  or fungicidal (echinocandins). The efficacies of these drugs depend on the quality of the hosts’ immune system, site and severity of infection, and pharmacokinetics of the drugs. Besides drug efficacies, therapeutic failures may result from evolution of drug resistance in the infecting Candida sp. The molecular mechanisms associated with antifungal resistance were summarized in this review. These mechanisms include overexpression of membrane transporters, altered cell wall and ergosterol biosynthesis, gain of function mutations in the transcription factors regulating membrane transporters, and ergosterol biosynthesis. The ongoing research on better understanding of these mechanisms may aid in detecting resistant isolates, identify novel drug targets, and inhibiting the rise of drug resistance.


Lycium chinense is an important edible and medicinal plant. Now, the complete chloroplast (cp) genome of L. chinense was assembled based on the Illumina sequencing reads. The cp genome of L. chinense was 155,736 bp long and contained two short inverted repeat regions , which were separated by a small single-copy region  and a large single-copy region . The cp genome encodes 113 unique genes, including 79 protein-coding genes, 30 transfer RNA genes, and 4 ribosomal RNA genes. The topology of the phylogenetic tree showed that L. chinense is closely clustered with species Lycium ruthenicum and Lycium barbarum. Lycium contains several well-known medicinal and food plants in Solanaceae, such as Lycium barbarum, Lycium chinense, and Lycium ruthenicum. There are about 80 Lycium species around the world  genome of L. chinense. The resultant data have been made publicly available as a resource for genetic information for Lycium species, and will provide a valuable plastid genomic resource for the future genetic and phylogenetic studies about L. chinense.The genus d Miller . Lycium  Olivier . The fruL. chinense were collected from the field of Kunming . The voucher specimen was deposited at Herbarium of Yunnan Agricultural University (No. 2019HSL003). Total genomic DNA was isolated from fresh leaves using a DNeasy Plant Mini Kit  according to the manufacturer’s instructions to construction cp DNA libraries. The Illumina sequencing was conducted by Biomarker Technologies Inc. . Resultant clean reads were assembled using GetOrganelle pipeline (https://github.com/Kinggerm/GetOrganelle). The genome was automatically annotated by using the CpGAVAS pipeline .The fresh leaves of L. chinense was 159,898 bp in length (Genbank accession number: MN102357). It was the typical quadripartite structure and contained two short inverted repeat regions(IRa and IRb) of 26,540 bp each, which were separated by a small single-copy (SSC) region  and a large single-copy (LSC) region . The cp genome encodes 109 unique genes, including 75 protein-coding genes, 30 transfer RNA (tRNA) genes and 4 ribosomal RNA (rRNA) genes. Twenty gene species are partially or completely duplicated, including nine PCG , seven tRNA , and all four rRNA . The overall GC content of the cp genome was 37.8%, while that of LSC, SSC, and IR regions was 35.9, 32.3, and 43.1%, respectively.The complete cp genome of Magnolia alba  as the outgroup. The combined datasets based on plastid genomes were aligned by MAFFT v7.307 (Katoh and Standley L. chinense has a close relationship with species L. ruthenicum and L. barbarum (A total of 28 cp genome sequences were selected to infer the phylogenetic relationships among the main representative species of Solanaceae with barbarum . The com


Persea americana Mill.) production contributes to the economic growth of East Africa. However, poor fruit quality caused by infestations of tephritid fruit flies (Tephritidae) and the false codling moth, Thaumatotibia leucotreta (Meyrick), hampers access to lucrative export markets. Remote sensing and spatial analysis are increasingly applied to crop pest studies to develop sustainable and cost-effective control strategies. In this study, we assessed pest abundance in Muranga, Kenya, across three vegetation productivity classes, viz., low, medium and high, which were estimated using the normalised difference vegetation index at a landscape scale. Population densities of the oriental fruit fly, Bactrocera dorsalis (Hendel) and T. leucotreta in avocado farms were estimated through specific baited traps and fruit rearing. The population density of T. leucotreta varied across the vegetation productivity classes throughout the study period, although not significantly. Meanwhile, B. dorsalis showed a clear trend of decrease over time and was significantly lower in high vegetation productivity class compared to low and medium classes. Ceratitis cosyra  was the most abundant pest reared from fruit with few associated parasitoids, Pachycrepoideusvindemmiae (Rondani) and Toxeumorpha nigricola (Ferriere).Avocado ( Agricultural landscapes contain a range of arthropod pests which influence crop productivity ,2,3. PesPersea americana Mill. (Lauraceae), is one of the most important horticultural crops grown in Kenya for subsistence, local markets and exports  . Two traps per farm, one for each species, were set 20 m apart along a 100 m transect. The transect ran in the middle of the farm, 30 m away from the edges and oriented from north to south. At each trap location along the transect, traps were hung at the east side of the tree, inside the upper 1/2 of the canopy and 2/3 outwards from the tree trunk. Insect samples were collected from traps every two weeks. The choice of this frequency was made to record population densities of a minimum of one generation, as the mean generation time is approximately 31 days and 48 days for B. dorsalis [T. leucotreta [Thaumatotibia leucotreta pheromone dispensers were renewed monthly while the ME coats were replaced after every six weeks. In the laboratory, insects were identified, counted and recorded. Keys described by De Meyer (1998) [Populations of dorsalis  and T. lucotreta , respectr (1998) , Drew & r (1998)  and Gillr (1998)  were useicipe’s Duduville campus, Nairobi  for processing and incubation. While in the laboratory, fruits were counted, weighed, placed into 10 L plastic containers  on a thin layer (0.5 cm) of sterilised sand and incubated in the laboratory . The lids of the containers were perforated at the middle and sealed with a fine mesh (1 mm) to provide ventilation. A total of 1030 pieces of fruit were collected during the study. Fruits were inspected every other day, and emerging pupae were transferred to Petri dishes. Emerged tephritid fruit flies, T. leucotreta and parasitoids were kept alive in the Petri dishes for four days to allow a full development of morphological features. Specimens were preserved in 90% ethanol for further identification.Avocado fruits were sampled monthly during the harvesting peak from February to May 2019. Eight pieces of fruit each from trees and the surrounding ground were collected per farm for further incubation in the laboratory. Four trees spaced at 20 m apart and located along the transect which had traps were targeted for fruit sampling. Fruits were randomly collected on the ground in a 2 m radius from the tree. Simultaneously, fruits with damage symptoms such as whitish perseitol exudates, holes and black spots were picked from the same tree. Fallen fruits were easier to find in some farms than in others because farmers usually feed livestock with fruits. In instances where the required number of fruits  were not found on the ground, only 2 to 4 pieces were collected. Tree and ground-collected fruits were kept separately inside 20 L × 12 W × 40 H cm brown paper bags and transported to Chalcidoid parasitoids were identified by Gerard Delvare, CIRAD, France ; John Noyes, The Natural History Museum, United Kingdom (Encyrtidae); Mircea-Dan Mitroiu, Alexandru Ioan Cuza University Iasi, Romania . The Nyssonidae specimen was identified by Robert Copeland using Bohart & Menke (1976) .B. dorsalis and T. leucotreta were expressed each as the daily capture per trap using the formula: total number of the insect caught/(number of serviced traps × number of days traps) [B. dorsalis and T. leucotreta as a function of landscape vegetation productivity and altitude. Collection time and farm were included as random effects. Full models were built with both random effects. Exceptionally for T. leucotreta, a correlation structure was added in the models to account for the spatial autocorrelation detected in T. leucotreta catches . Simplified models were defined from the full model by sequentially dropping the random effects and/or the correlation structure. The Restricted Maximum Likelihood approach was used to estimate model parameters and the best-fitting model was selected based on the lowest Akaike Information Criterion (AIC) value [P values were adjusted with the Benjamini-Hochberg method. Statistical analyses were performed in R version 3.6.3 [Catches of s traps) . The daiC) value . Model vC) value . The infon 3.6.3 , using ton 3.6.3 , emmeanson 3.6.3  and multon 3.6.3 . SignifiB. dorsalis (99.6%). Few other fruit flies such as C. cosyra, Ceratitis  cuthbertsoni (Munro), Ceratitis  ditissima (Munro), Perilampsis pulchella (Austen) and Ceratitis (Pterandrus) fasciventris (Bezzi) were also caught. The false codling moth T. leucotreta was the most abundant moth species (99.8%) caught in the delta trap followed by the fall armyworm Spodoptera frugiperda (Smith) (0.2%).Avocado farms in low and medium vegetation productivity classes were located between 1329 m and 1689 m asl, while farms in the high class were located between 1563 m and 2020 m asl. The most abundant fruit fly caught in ME-baited trap was B. dorsalis varied between vegetation productivity classes throughout the season. In the low vegetation productivity class, there were two peaks of B. dorsalis catches, respectively in March and May 2019 (up to 25 flies/trap×day). After May 2019, catches of B. dorsalis started declining until the end of the study in August 2019 with 5 flies/trap×day  throughout the study. Slight increases of 6–7 flies/trap×day were recorded in mid-May and mid-June 2019 . Analysis indicated that the farm  and collection time  influence the variance of B. dorsalis catches. Landscape vegetation productivity significantly influenced B. dorsalis catches , being higher in medium vegetation productivity class, followed by low and high classes (The best-fitted model which explained  classes .T. leucotreta remained more abundant in the high vegetation productivity class (up to 6 moths/trap×day in May 2019) than in the low and medium classes, except in June and August 2019, during which the captures were more abundant in low and medium vegetation productivity classes than high class . The best fitted model for T. leucotreta included both farm and collection time as random effects (AIC = 1979.99). The variation among farms  and collection time  explained the variance of T. leucotreta catches. However, there was not a significant effect of landscape vegetation productivity  on the daily capture of T. leucotreta . The daily capture of the pest decreased by 0.05 for every increase unit in altitude. In contrast, T. leucotreta did not vary with altitude .Analysis showed that altitude strongly influenced T. leucotreta and parasitoids were recorded during fruit incubation, even though 7% of pupae failed to hatch. Ceratitis cosyra was the most abundant species (97.7%) recovered from fruits, followed by B. dorsalis (1.6%) and T. leucotreta (0.3%). The analyses focused only on the infestation index for C. cosyra because of its high prevalence. The C. cosyra infestation index of ground-collected fruit was significantly different across vegetation productivity classes  .The mean mass per fruit varied between 150 g and 190 g. Fruit flies, = 0.043) . Fallen Epitranus sp. nov. , Cheiloneurus sp. , Pachycrepoideus probably vindemmiae (Rondani) , Toxeumorpha probably nigricola (Ferriere)  and Bembecinus sp.  (Nyssonidae) (Pachycrepoideus vindemmiae (the first record in Kenya) and T. nigricola emerged from C. cosyra pupae. Host pupae of Cheiloneurus sp., Epitranus sp. nov and Bembecinus sp. were not identified. Epitranus sp. nov. was found only in the medium vegetation productivity class, while Cheiloneurus sp., P. vindemmiae, T. nigricola and Bembecinus sp. were recorded in high vegetation productivity class. No parasitoid was recorded in low vegetation productivity class. Only one individual Cheiloneurus sp. was reared from tree-collected fruits in the medium vegetation productivity class.Only 11 chalcidoid and one sphecoid wasps were recovered during fruit incubation. They were identified as sonidae) . Among tB. dorsalis was caught in ME-baited traps in the avocado farms throughout the season. Landscape vegetation productivity strongly influenced B. dorsalis populations. Bactrocera dorsalis catches were more abundant in medium and low than high vegetation productivity class. This involves that low and medium vegetation productivity are high-risk infestations areas and should receive considerable management efforts. Although vegetation productivity is a general measurement and does not reflect specific resources for the pest, it may probably have interacted with the presence of alternative host plants and influenced the distribution of B. dorsalis. Indeed, there was a gradual decrease in B. dorsalis captures over time across the three vegetation productivity classes. This trend is likely related to the phenology of avocado and alternative host plants such as mango. For instance, the presence of fruiting mango trees in and around avocado farms in low and medium vegetation productivity classes, as observed during the field sampling, would have also contributed to sustaining B. dorsalis populations, as mango is a primary host of B. dorsalis [B. dorsalis catches decreased with increase in altitude. This study further supports the characterisation of B. dorsalis as a lowland pest [Bactrocera dorsalis populations in avocado farms decrease at higher elevations in Kenya and Tanzania [B. dorsalis population in avocado farms, but also the altitude. Future studies should specifically look at the influence of the abundance, distribution and diversity of host plants.The fruit fly dorsalis ,27. Alsoand pest ,50. BactTanzania . TherefoB. dorsalis, we found no significant effect of vegetation productivity and altitude on T. leucotreta captures. However, T. leucotreta was higher in high than low and medium vegetation productivity classes early in the season. Daily captures of T. leucotreta in low and medium vegetation productivity classes showed an increase in July 2019, probably because of the presence of alternative hosts such as macadamia and coffee which would have supported T. leucotreta populations when avocado trees were flowering and had few fruits. The trapping of T. leucotreta for a longer period (at least two years) might have given a clearer and consistent temporal trend of its population density across the landscape vegetation productivity classes. The lack of altitude effect on T. leucotreta can be explained by the thermal tolerance of the pest. Populations of T. leucotreta are sustained well with the presence of host plants in low and highlands areas in South Africa [T. leucotreta host plants need to be further investigated.Unlike h Africa  and Easth Africa . This reC. cosyra was the most abundant species reared from avocado. A potential explanation to this could be the high preference of B. dorsalis for mango over avocado in our study sites. B. dorsalis also prefers mango over Citrus sp., cucurbits and papaya [C. cosyra by B. dorsalis on mango [C. cosyra could have been displaced by B. dorsalis on mango and sought refuges in avocado. This involves fruit fly management in avocado farms also targeting mango trees in the farms. In future studies more emphasis will be put on the monitoring of C. cosyra with terpinyl acetate baited traps. Bactrocera dorsalis was the dominant species reared from avocado in Taita Hills (Kenya), a region where mango is rare, thus making avocado the main host plant for B. dorsalis [Thaumatotibia leucotreta rarely emerged from ground-collected fruits. Females lay eggs on the fruit skin, and larvae hatch and tunnel inside the fruit. Exceptionally on avocado, larvae exit the fruit and pupate in a silken cocoon in the soil [Unexpectedly, the mango fruit fly d papaya . This pron mango . In our dorsalis . Thaumatthe soil . During Bactrocera dorsalis, C. cosyra and T. leucotreta rarely emerged from tree-collected fruit, although infestation symptoms were recorded. Hanging and unripe avocado fruits develop defence mechanisms such as encapsulation and antibiosis to prevent larval development of fruit flies and T. leucotreta [T. leucotreta were more abundant in ground-collected fruits than tree-collected fruits. Fruit fly species C. cuthbertsoni., C. ditissima, P. pulchella and C. fasciventris caught in ME Lynfield traps were not found during the incubation of fruits. Therefore, these species likely are not pests of avocado and could have inadvertently trapped and killed by the malathion insecticide contained in the para-pheromone.ucotreta ,24. AlsoP. vindemmiae and T. nigricola emerged from C. cosyra pupae. Pachycrepoideus vindemmiae is an idiobiont ectoparasitoid used in Hawaii, South America and Benin for the biological control of drosophilids and tephritid fruit flies such as Bactrocera cucurbitae (Coquillet), Bactrocera latifrons (Hendel), C. capitata and B. dorsalis [Toxeumorpha nigricola is commonly found in Africa parasitising pupae of leaf-mining Lepidoptera and Diptera such as C. capitata [Toxeumorpha nigricola may have a lower preference for C. cosyra than other hosts. Moreover, P. vindemmiae is a facultative hyperparasitoid attacking parasitised fruit flies [Epitranus sp. and encyrtids such as Cheiloneurus sp. are occasionally reared from flies and may have parasitised drosophilids, calliphorids or muscoid flies which emerged during fruit incubation [Bembecinus sp. like most of the sphecoids are mud daubers and would have been collected in the ground at the larval or pupal stage during the incubation of fallen avocado fruits. Further studies will help shed light on the host utilisation and efficiency of these parasitoids.In this study, five parasitoids species were reared from avocado, among which dorsalis ,64. Toxecapitata . Howevercubation ,67. BembB. dorsalis and C. cosyra and the false codling moth T. leucotreta damage avocado fruits in Kenya. The vegetation productivity determined by NDVI influences catches of B. dorsalis but not T. leucotreta. Similarly, B. dorsalis populations increase with a decrease in altitude, while T. leucotreta do not vary with altitude. Landscape vegetation productivity is also a predictor for C. cosyra infestation of avocado fruits. In this study, very few parasitoids emerged during fruit incubation, among which the pteromalids T. nigricola and P. vindemmiae parasitised C. cosyra.Fruit flies


Rhodococcus spp. from Tunisia are reported here. Two Rhodococcus fascians strains were obtained from almond rootstocks, and one Rhodococcus kroppenstedtii strain was obtained from a pistachio tree. The fourth Rhodococcus sp. strain was isolated from an ornamental plant.The draft genome sequences of plant-associated  Rhodococcus spp. from Tunisia are reported here. Two Rhodococcus fascians strains were obtained from almond rootstocks, and one Rhodococcus kroppenstedtii strain was obtained from a pistachio tree. The fourth Rhodococcus sp. strain was isolated from an ornamental plant.The draft genome sequences of plant-associated  Rhodococcus spp. are known to cause disease on herbaceous and woody species . Protein-coding features and tRNA were predicted using Prodigal version 2.6 ( strains  were iso strains  and grow strains . Genomic strains  with a s strains . The ass strains . The gension 2.6 , and rRNsion 2.6 . For taxsion 2.6 . DefaultRhodococcus species. The sequence of the isolate K5 genome is 97.9% identical to the type strain of Rhodococcus kroppenstedtii, the isolate B10 genome is identical to multiple Rhodococcus type strains, the isolate SB10 genome is 97.7% identical to Rhodococcus fascians NBRC 12155, and the isolate GS6 genome is 97.8% identical to Rhodococcus fascians NBRC 12155.The assembly statistics and total number of genes are shown in JAAFYU000000000, JAAFYV000000000, JAAFYW000000000, and JAAFYX000000000. The versions described in this paper are JAAFYU010000000, JAAFYV010000000, JAAFYW010000000, and JAAFYX010000000. The raw sequence reads have been deposited in the NCBI Sequence Read Archive under BioProject number PRJNA598862 and run numbers SRR11109605, SRR11109606, SRR11109607, and SRR11109608.This whole-genome shotgun project has been deposited at DDBJ/ENA/GenBank under accession numbers


Candida albicans is a common opportunistic pathogen that can cause serious infection by blood transmission. C. albicans enters the blood circulation and adheres to the endothelial cells of the vascular wall. However, the detailed mechanism of the effect of C. albicans on the endothelial cells remains unclear. In this study, the microarray expression profile of human umbilical vein endothelial cells exposed to C. albicans was analyzed. The 191 up-regulated genes were enriched in TNF, T cell receptor, and NF-kappa B signaling pathways. The 71 down-regulated genes were enriched in pyruvate metabolic, purine nucleotide metabolic, purine nucleotide biosynthetic, and humoral immune response processes. Gene set enrichment analysis showed that apoptosis, oxidative phosphorylation, IL6/JAK/STAT3 signaling pathways were enriched. Moreover, two hub genes with a high degree of connectivity, namely, MYC and IL6, were selected. Molecular screening of traditional Chinese medicine libraries was performed on the basis of the structure of MYC protein. The okanin had the highest docking score. MYC might be used as molecular targets for treatment. In addition, okanin may inhibit the infection of C. albicans. Thus, MYC can be subjected to further research.The online version of this article (10.1007/s12088-019-00847-5) contains supplementary material, which is available to authorized users. Candida albicans is a polymorphic yeast and one of the most important pathogens that causes iatrogenic infections in immunodeficient populations [C. albicans can cause mucosal infections, thereby causing diseases such as thrush, fungal vaginitis, and rash [C. albicans can invade the mucosal epithelium and vascular endothelium, thereby causing disseminated infections in susceptible populations [C. albicans invades the host’s non-phagocytic cells, such as endothelial cells, and plays a vital role in the early stages of the disease [C. albicans on endothelial cells.ulations . In geneand rash , 3. In sulations . In the  disease . CurrentBidens [C. albicans infection activity of okanin has not been investigated yet.High-throughput virtual screening revealed that okanin has the highest docking score with MYC , 7. OkanBidens . PharmacBidens . AlthougBidens , the antC. albicans infection were analyzed by performing bioinformatics analysis. The key functions and pathways of endothelial cells after C. albicans infection were also identified. By conducting the topological analysis, we found and further screened the key hub genes in the affected endothelial cell lesions to obtain okanin. The candidate molecule for traditional Chinese medicine (TCM) may inhibit C. albicans infection.In the present study, differentially expressed genes (DEGs) of endothelial cells after C. albicans-infected HUVEC monolayer. The data quality was assessed by calculating residuals.sign, residuals, weight, relative log expression (RLE), normalized unscaled standard errors (NUSE), and RNA degradation. We used the R packages of pheatmap and limma to analyze the difference in RNA expression profiling among the eight groups. We set the |log2fold change| ≥ 0.6 of the cutoff limit for DEGs.The RNA expression profiling of GSE7355 downloaded from the GEO database was used in this study. Four independent experiments compared the expression profiles of untreated HUVEC monolayer and of GSEA was conducted using the GSEA software .http://metascape.org). The enriched pathways or functions were then drawn into bubble maps with the R package of ggplot2. PPI annotation of the DEGs was retrieved from the STRING database. The PPI network was visualized using the Cytoscape software. Then, the Cytoscape apps of CentiScape and MCODE were used for detecting densely connected regions [The DEGs were further analyzed using the gene ontology (GO), KEGG pathway, and protein–protein interaction (PPI) analyses . The GO  regions , 14.http://tcm.cmu.edu.tw/) [http://www.rcsb.org/, PDBID: 1NKP) [A total of 32,364 TCM molecules were obtained from the TCM database (edu.tw/) . All theedu.tw/) , 17. ForD: 1NKP) . The proD: 1NKP) , 20.The HUVEC cell lines, obtained from KeyGen Biotech , were transfected with dual-reporter constructs of MYC using the transfection regents. After transfection of 24 h, cells were treated with 40 µM of okanin. After another 48 h, the culture medium was collected into a 96-well plate and the luminescence was measured using a luminometer. The fluorescence intensity reflects the transcriptional activity.The okanin was added to the HUVEC cell culture at a concentration of 40 µM when the cell density reached to 80%. After incubation of 48 h, the cells were lysed and 30 µg of total protein was used for sodium dodecyl sulfatepolyacrylamide gel electrophoresis and transferred onto polyvinylidene difluoride membranes . The membrane was incubated with primary antibodies against GLUT2 , MYC , and GAPDH , followed by the secondary antibody . The blots were detected using an enhanced chemiluminescence detection kit . The expression analysis was performed using the ImageJ software. The ratio of densitometry value to the corresponding GAPDH was used to reflect the relative protein expression.C. albicans on endothelial cells, we downloaded the GEO database of GSE7355 from the GEO database. By calculating residuals.sign, residuals, weight, RLE, NUSE, and RNA degradation, the data quality was acceptable . The heat map analysis result showed a quite different expressed gene profile between the control HUVEC monolayer and C. albicans-infected HUVEC monolayer . The volcano plot further described a difference in gene expression profile between the two groups. For instance, many genes were considerably upregulated and labeled in red color, whereas some genes were remarkably downregulated and labeled in blue color .To clarify the effects of C. albicans infection could affect the apoptosis and oxidative phosphorylation processes. Furthermore, the C. albicans infection could affect the inflammatory response and its related signaling pathways, namely, NFKB, IL6-JAK-STAT3, and IL2-STAT5 pathways . Further analysis of the DEGs revealed that C. albicans infection extensively promoted immune- and inflammation-related signaling pathways, such as TNF, Toll-like receptor, T cell receptor, NF-kappa B, and chemokine-mediated signaling pathway . This network consisted of 105 nodes and 360 edges , may inhibit C. albicans infection by inhibiting MYC. Okanin can lower the mRNA and protein level of IL6 in the LPS-induced pro-inflammatory model [C. albicans infection.This study demonstrated that okanin, a major effective constituent of ry model . It alsory model . The strry model . TherefoBelow is the link to the electronic supplementary materialSupplementary Material 1Supplementary Material 2


Sarcophaga (Pandelleisca) mersinensissp. nov. is described from the Mediterranean region of Turkey. The male terminalia are documented with line drawings, photographs and scanning electron microscope images. The species is compared with the two most similar species, Sarcophaga (Pandelleisca) baudeti (Lehrer) and Sarcophaga (Pandelleisca) theodori (Lehrer), both known from Israel. A key is provided to the western Palaearctic species of Pandelleisca Rohdendorf.A new species,  Pandelleisca Rohdendorf, 1937  contains 24 species of flesh flies, mainly distributed in the Oriental region (S. (P.) baudeti  and S. (P.) theodori , both known only from Israel, and S. (P.) similis Meade, 1876, which is widely distributed in both the Palaearctic and the Oriental regions (Sarcophaga (P.) similis is the only representative of the subgenus recorded in Turkey so far  from the Mediterranean region of Turkey, providing photographs, scanning electron microscope (SEM) images, and line drawings of the male terminalia, and a key to the western Palaearctic species.This paper describes a new species of The material was collected during the years 2013–2017 in Mezitli and Erdemli districts of Mersin Province of Turkey using insect sweep nets. The specimens were killed in ethyl acetate vapour, pinned shortly afterwards when they were still fresh and air-dried.Males were relaxed in a humidifier, and the terminalia of each specimen were detached from the abdomen using forceps and fine insect pins. The dissected terminalia of the holotype were subjected to 10% KOH for 12 hours, rinsed with distilled water and placed into glycerine for further examinations under a Leica S8APO stereomicroscope.SEM by fixing on an aluminium stub with carbon double-stick tape. The gold-coated specimens were examined and imaged in a FEI Quanta 450 FEG scanning electron microscope at BILTEM (Science and Technology Application and Research Centre of Yozgat Bozok University) using high vacuum.The air-dried genitalia of the paratype were prepared for Light microscope photographs were taken with a Leica DFC 450 camera integrated on a Leica M125 stereomicroscope and stacked in Helicon Focus Pro (version 7.6.1). Line drawings of terminal structures were produced with CorelDraw Graphics Suite 2019.The terminalia of the holotype of the new species are stored in a micro-vial with glycerine and the dissected parts of the terminalia of the paratype were glued to a piece of card and both are pinned together beneath the source specimens. All the samples are deposited in the Entomology Collection of the Department of Plant Protection, Faculty of Agriculture, Bozok University, Yozgat, Turkey.S. (P.) baudeti  and S. (P.) theodori  with the other western Palaearctic species were based on the original descriptions of For identification, the following works were consulted: Data from labels of the type specimens are quoted verbatim: commas are used to separate the lines on the same label, labels are separated by a double forward slash, and any remarks are given in square brackets.Taxon classificationAnimaliaDipteraSarcophagidae1736C6B6-E816-5E24-87F6-D9AD463A4187http://zoobank.org/31EF981D-0E7D-459D-98F7-F4D6815DABC3Holotype: ♂, TR// Mersin province [southern Turkey], Mezitli district, 1.2 km NE Kuzucu village, 608 m, 36°50'32"N, 34°25'24"E, 07.VII.2017, Leg. G. Bakır [printed on white paper] // Holotype ♂ Sarcophaga (Pandelleisca) mersinensis, Det. Pekbey, 2020 [printed on red paper]. Paratype: ♂, TR// Mersin province [southern Turkey], Erdemli district, Kösbucağı village, 542 m, 36°40'58"N, 34°14'37"E, 11.VII.2013, Leg. C. Metin [printed on white paper]. Sarcophaga (Pandelleisca) mersinensis, Det. Pekbey, 2020 [printed on red paper].Sarcophaga (Pandelleisca) mersinensis sp. nov. is similar to the East Mediterranean species S. (P.) theodori  and S. (P.) baudeti . It is distinguished from S. (P.) baudeti by having a brown epandrium  theodori by the following features of the male terminalia: in S. (P.) mersinensis the harpes are subtriangular in lateral view, poorly sclerotized and very small .Head. Black with golden microtomentum and the eye 0.38 times as wide as head in dorsal view. Inner vertical seta long and strong, outer vertical and proclinate orbital seta indistinct. Reclinate orbital seta well developed. Eye bare. Postocular seta black, arranged in two rows on each side of occiput. Frons apically protruding and at its narrowest point 0.68 times as wide as an eye in dorsal view. Frontal vitta black, slightly widening to antennal insertion, 0.48 times as wide as frons. Frontal bristles 11 or12 pairs, not descending below of the midline of pedicel. Parafacial plate black with golden microtomentum, with a row of fine and black setulae in lower half near eye margin. Parafacial at its narrowest point 0.42 times as wide as an eye at maximum eye width in lateral view. Gena black with golden-silvery microtomentum, anterior half covered with black seta, post genal seta pale. Gena in profile 0.36 times as high as the height of an eye. Genal dilation distinct, brownish black. Vibrissa well developed. Facial ridge with a few decumbent setulae above vibrissa. Antenna brownish black, pedicel with a reddish-brown tinge on the distal part. Postpedicel 2.76 times longer than pedicel. Arista light brown, 2/3 plumose, slightly thickened on basal part. Prementum and palpus dark brown, 2.2 times longer than wide , katepisternals 2 + 1; scutellum with two pairs of subapical setae, one pair of basal and one pair of discal setae.Legs black. Fore tibia with three anterodorsal and one posteroventral seta. Mid femur with scarce and short ctenidium. Mid tibia with two or three anterodorsal, one anteroventral, and three or four posteroventral setae. Hind tibia with a row of hair-like setae on posteroventral and ventral surface, with two strong anterodorsal setae, 1 anteroventral and one posterodorsal.Wing. Hyaline. Epaulet black. Basicosta bright yellow. Costal spine absent. Vein R1 bare. Vein R4+5 dorsally with short and black setulae at base. Distal part of M curved at a right angle. Second costal section 1.44 times as long as fourth costal section. Cell r4+5 open at wing margin. Haltere brown. Lower calypter yellowish white .The species epithet is derived from Mersin Province situated in the Mediterranean region of Turkey, where the type series was collected.Pandelleisca are recorded only from the Oriental region, and the six Palaearctic species are restricted to the far eastern territories including Palaearctic China, Russia, North Korea, South Korea, and Japan (Sarcophaga (P.) similis is widely distributed throughout the Palaearctic and Oriental regions and is found mostly in mesophytic forest habitats (S. (P.) mersinensis was collected from the Mediterranean coastal region as are the two Israeli species, S. (P.) theodori (S. (P.) baudeti .Thirteen species of nd Japan . Sarcophhabitats . The spehabitats . The newtheodori  and S. (Sarcophaga (P.) theodori is the species most similar to S. (P.) mersinensis sp. nov. with regard to morphological structures of the phallus. In both species, the distiphallus expands abruptly apicolaterally in dorsal view and has elongated ventral appendages. The median process of juxta is broad, flattened, and spur-like in lateral view in both species, and bends anteroventrally with a wide angle towards the lateral juxtal arms. These arms are paired, widened basally, and slightly bifurcated at the tip. The superior vesical lobes are leaf-like, and the inferior ones are relatively thin and spiky. The lateral styli are broad and tubular in both species.Pandelleisca species have a shiny black epandrium (S. (P.) ballardi , S. (P.) brachiata , S. (P.) quinqueramosa , and S. (P.) theodori  the colouration of the epandrium is brown as in S. (P.) mersinensis sp. nov.As stated by some authors, the vast majority of pandrium , but in Pandelleisca and Liosarcophaga are still unsettled. Some molecular studies have shown that Liosarcophaga and Pandelleisca are not monophyletic in their current circumscriptions, and that the subgenus Pandelleisca as proposed by Pandelleisca. The phylogenetic placement of S. (P.) similis in the study of Sarcophaga (Rosellea) aratrix Pandellé rather than grouping with all other species here considered under Pandelleisca (Sarcophaga.The taxonomic limits between elleisca . This muPandelleisca species appear to be recorded from the coastal geographic regions in Turkey, it is thought that future more comprehensive faunistic surveys will reveal the true distribution of this subgenus.In conclusion, although all


Aedes aegypti, Anopheles minimus and An. dirus, and two field populations of An. harrisoni and Ae. aegypti, respectively. Established TFT sublethal (LC50 and LC75), lethal concentrations (LC99) and discriminating concentrations (DCs) were selected corresponding to each mosquito test species. Spatial repellency and contact excitation (‘irritancy’) responses on adult mosquitoes to TFT were assessed using an excito-repellency assay system. At LC50, TFT exhibited strong avoidance with An. minimus (60.1% escape) and An. dirus (80% escape) laboratory strains, showing between 12 and 16x greater escape response than Ae. aegypti (5% escape). Repellency responses for field collected Ae. aegypti and An. harrisoni were 54.9 and 47.1% escape, respectively. After adjusting the initial contact escape response (a measure of combined irritancy and repellency) to estimate only escape due to contact, the LC50 and LC99 showed moderate escape irritancy with laboratory Ae. aegypti (41.4% escape) and no contact activity against the field population. Adjustment showed only weak contact activity (16.1% escape) in laboratory An. minimus at LC50. Spatial repellency is the predominant mode of action of TFT among colonized and field mosquitoes used in this study. Established baseline (susceptible) dose-response curves assist in optimizing SR products for mosquito control and pathogen transmission prevention.Airborne spatial repellency (SR) is characterized and distinguished from other chemical actions including contact locomotor excitation and toxicity. The use of volatile spatial repellents is a potential new intervention class for combatting mosquito-borne pathogen transmission; therefore, continuing investigations on the actions of these chemicals that modify mosquito host‐seeking behavior  is needed. The objective of this study is to characterize the key behavioral avoidance actions of transfluthrin (TFT) to advance spatial repellent development into practical products. Behavioral avoidance responses were observed for adult laboratory strains of  Aedes-borne viruses  are projected to expand and pose a greater risk, both geographically and demographically, in the decades ahead [Plasmodium vivax infections, a latent relapse form of parasite complicating elimination goals in the country [More than 4 billion people, primarily residing in tropical and sub-tropical countries, are at risk of infection from mosquito-borne diseases. For example, globally, an estimated 390 million dengue infections occur and around 200 million malaria cases are reported annually , 2. Denges ahead . While mMosquito vector control through the use of insecticide-treated bed nets (ITNs) and indoor residual spraying (IRS) are considered to have made major contributions towards the reduction in the global malaria burden since 2000 . Among tAedes aegypti (L.) and Aedes albopictus (Skuse) are the two most important vectors of dengue as well as chikungunya and Zika viruses [Aedes aegypti, in particular, is a formidable mosquito to control using current vector abatement methods. It is a strongly synanthropic/anthropophilic species and predominantly found in and near human dwellings with a high tendency of adults to rest inside houses [ viruses . In gene viruses . Aedes ae houses .Anopheles malaria vector species in Thailand, Anopheles dirus Peyton & Harrison and An. minimus Theobald are among the most important [Among the primary mportant , 15. Botmportant . Althougmportant , 17. Premportant , 14, 18 mportant .Ae. aegypti (37.5% repellency) at 0.001% and Ae. albopictus (45% repellency) at 0.01%. However, to our knowledge, there have been no published studies with TFT distinguishing the two types of behavioral avoidance responses (irritancy and repellency). Information on TFT and mosquito responses are limited; therefore, investigations are required on this AI for development of innovative spatial repellent products.Therefore, there has been increased interest in the spatial repellent properties of certain AIs as a possible means of arresting outdoor biting –22. By dA better understanding of the functional effects of chemical insecticides and mosquito responses is, therefore, critical in designing practical interventions of TFT-based spatial repellent products , 26. TheAedes aegypti (L.) (USDA strain), obtained from the U.S. Department of Agriculture, was maintained at the Department of Entomology, Faculty of Agriculture, Kasetsart University (KU) for over 20 years. 2) Anopheles minimus s.s. Theobald (DDC strain), from the Malaria Division, Department of Disease Control, Ministry of Public Health, Nonthaburi, Thailand, originally collected in Rong Klang District, Prae Province, northern Thailand in 1993 and maintained at the Kasetsart University laboratory shortly thereafter. 3) Anopheles dirus s.s. Peyton & Harrison (TMMU strain) was originally collected in Khao Mai Kaeo Sub-district, Bang Lamung District, Chonburi Province, eastern Thailand in 1981 and obtained from the Department of Medical Entomology, Faculty of Tropical Medicine, Mahidol University in 2016. This strain has been maintained continuously at KU insectary since 2016. Mosquito colonies were reared following standard handling procedures and conditions  at the KU laboratory [ad libitum with 10% sucrose (w/v) solution as sustenance. An artificial membrane feeding technique [An. dirus; all other species were self-mating in holding cages.Three pyrethroid-susceptible laboratory strains and two recently collected field populations of adult mosquitoes were used. Laboratory strains: 1) boratory , 30. Larechnique  using huechnique  was requAedes aegypti field population was collected as immature stages from Muang District, Nonthaburi Province, central Thailand  in 2016 and maintained at KU insectary; 2) Anopheles harrisoni Harbach & Manguin (a species member of the Minimus Complex) field population was collected from Pu Teuy Village, Sai Yok District, Kanchanaburi Province  according to Sungvornyothin et al. [1 to F3 generation mosquitoes.Field populations: 1) n et al. . The assn et al.  prior ton et al. . All ER R,3S)-3--2,2-dimethylcyclopropanecarboxylate; 97.90% purity) was used. Transfluthrin stock solutions were prepared using analytical grade acetone  as an organic solvent and silicone oil (Dow Corning® 556 cosmetic grade) as a carrier at a ratio of 1.91:0.95.Technical grade TFT  and a discriminating concentration (DC) based on doubling the LC99 x 2 for Ae. aegypti, An. minimus and An. dirus [Whatman No. 1 filter papers, measuring 14.7 x 17.5 cm, were uniformly impregnated with 2.8 mL of TFT solution using a calibrated micropipette. Papers were treated with established 50%, 75% and 99% lethal concentrations  were introduced into each of four chambers . Following a 3-min ‘acclimation’ period, the exit portal was opened for each chamber to initiate observations (escape). The number of mosquitoes that escaped from each exposure chamber into the attached receiving cage were recorded at 1-min intervals for a period of 30 min. At post-exposure, all mosquitoes were transferred to holding cups labeled by the chamber and movement response (escape or non-escape). The number of dead or knockdown (moribund) mosquitoes were recorded separately from inside (those remaining) and outside (having escaped) each chamber. All live and moribund mosquitoes in the chamber were held separately and provided with cotton-soaked 10% sugar solution for 24-h before recording the final mortality determination. The ambient air temperatures and relative humidity were recorded at the beginning of each 30-min trial period. All trials were performed during daytime hours (09:00–16:30 h) and each trial series were replicated four times. All assays were carried out in the same laboratory space, under identical conditions and ER assay systems for both colonized and field-collected mosquitoes.The percent ER escape responses and 24-h mortality were adjusted using Abbott’s formula when escape or mortality in the paired controls (untreated chambers) was between 5 and 20% of the test sample . The num25), 50% (ET50) and 75% (ET75) of test population to escape was estimated. A log-rank test [P < 0.05).Using initial escape data (Abbott’s formula adjusted only), Kaplan-Meier survival analysis was used to estimate rate of mosquito escape in contact and noncontact test formats, and for comparing differences in mosquito escape between the four concentrations . The timank test  comparedAe. aegypti, An. minimus and An. dirus using different species-specific concentrations are shown in Tables P < 0.05) greater percent escape in both noncontact and contact trials compared to the controls, except for Ae. aegypti in noncontact at LC50 (P = 0.0807) (P > 0.05) (Ae. aegypti (USDA), LC50 to LC99 produced percent escape in contact tests (Abbott’s adjusted percent only) ranging between 48 and 66% (99 (0.03412%) and the DC (0.06824%) due to the higher knockdown (KD) effect inside the chamber  compared to those that escaped (4.8–24% KD). Conversely, a positive relationship was found in noncontact test as percent escape is increasec as TFT concentration increased (Tables The contact excitation and noncontact repellency effects of TFT on laboratory-reared  0.0807) . No sign > 0.05) . For Ae. and 66% . The per(99 0.0342% and thAn. minimus (DDC), LC50 and LC75 TFT exhibited the highest mean percent escape in both contact and noncontact tests, ranging from 85.7–80.8% (contact) and 60.1–66.5% (noncontact), respectively (50 and 43.4% at the highest concentration (DC). Relatively high repellent activity in noncontact test was observed with An. dirus (TMMU) ranging from 46.4% escape (LC75) to 85.9% escape (LC99), while the contact test showed a final percent escape ranging between 54.2% (DC) and 74.1% (LC50) across all four concentrations  and contact (ET75 = 25 min) tests. Overall, at all concentrations, at least 50% of An. minimus and An. dirus escaped TFT-treated chambers in both trials within 6–12 and 3–15 min (ET50), respectively. In contrast, Ae. aegypti showed the least excito-repellency, both in final lesser percent escape and longer length of time required to escape the chamber.A wide range of escape time (ET) activity was observed between species and concentrations in noncontact and contact tests . In onlyAe. aegypti, unadjusted contact escape was significantly stronger than noncontact repellency between LC50 and LC99, but not the DC  in non-escape mosquito, while no knockdown (0%) and low to moderate %KD (4.8–36.8%) was found in escape group for all concentration in both contact and noncontact tests  and escape (63.6–83.3%) mosquito, respectively (An. dirus in both ER assays (High percentage of knockdown (82.5–94.7%) was only observed against ct tests . For An.ectively . WhereasR assays . Howeverape 62.5–00% and e %KD 4.8–6.8% was Ae. aegypti had relatively lower escape rates in noncontact tests at all concentrations, and more moderate responses in the contact tests  for escape data, the survival curves at 1-min intervals in contact and noncontact tests were generated Figs . Comaprect tests . Conversats Figs  and 3.Ae. aegypti exhibited only moderate behavioral avoidance (between 26.5–43.3% escape) with contact exposure at LC50—LC99, and only contributed 9.5% of the escape at the highest concentration (DC)  . However minimus  and 10% n. dirus . These fAe. aegypti showed no significant differences in escape responses except for LC50- LC99 and LC99-DC pairings. For noncontact tests, all LC50 pairings with the other three higher concentration were significantly different. For An. minimus, significant differences in contact escape were found in four pairwise comparisons (excluding LC50-LC75 and LC99-DC); whereas there was no difference in escape between noncontact comparisons. For An. dirus, no significant differences were observed in contact escape pairings and in only one instance was a difference detected in the noncontact assays (LC75-LC99).Within species, log-rank multiple paired comparisons between TFT concentrations and escape responses in contact and noncontact tests were performed . In contAe. aegypti (NON) were found resistant to TFT  using the previously established DC (0.06824%); whereas, An. harrisoni (KAN) was fully susceptible to TFT using the DC 0.06382% [WHO insecticide susceptibility tests showed that field-collected 0.06382% .50) established for Ae. aegypti (USDA) (0.00852%) and An. minimus (DDC) (0.00423%) [Ae aegypti (NON) and An. harrisoni (KAN), respectively. In noncontact tests, Ae. aegypti (NON) had significantly higher escape response compared to the USDA colony . There was a significantly higher percent escape in An. minimus (DDC) compared to An. harrisoni (KAN) in both noncontact (P = 0.0048) and contact (P < 0.0001) tests  (P < 0.0001) reductions in estimated percent escape due to contact only was also seen in the colonized strains. High % knockdown (>93.6%) were also observed in non-escape An. harrisoni KAN for both contact and noncontact tests, while 68–74% knockdown was found for non-escape Ae. aegypti NON mosquito. Both field populations recovered  after 24-h post exposure with reduced mortality ranging from 0% to 20%   were useP < 0.000; 1 and 63%  were contact) . Followi% to 20% .Ae. aegypti, An. minimus and An. dirus. When allowing mosquitoes a free flight choice, TFT was most effective in eliciting substantial spatial repellency, particularly Anopheles mosquitoes. Relatively high escape reactions in both contact and noncontact tests with all concentrations, lethal and sublethal, occurred with An. minimus and An. dirus. Aedes aegypti showed a more reduced and delayed escape responses compared to the anophelines. Transfluthrin at LC50, LC75, and LC99 also induced a moderate knockdown effect and 24 h mortality against Ae. aegypti, but higher KD and mortality on anophilines, particularly An. dirus. At the final concentrations used, the lower post-exposure (24 h) mortality compared with the higher initial percentage of knockdown mosquitoes at 30 min , for those not escaping (remaining in the exposure chamber) and those successfully escaping the chambers, suggests that the overall behavioral avoidance was the primary mode of action of TFT.The key feature of the excito-repellency (ER) assay system is the separation between contact excitation (‘irritancy’) and noncontact spatial repellency of a target compound that result in movement away or non movement from the physical source of the active compound (i.e. impregnated papers) ,44. UsinPreviously, the standard WHO susceptibility assay, using lethal and discriminating concentrations of TFT on cellulose filter papers, was used to measure phenotypic response of mosquitoes with survival or mortality as endpoints . Given tThe relatively high vapor pressure chemistry of TFT compared to other pyrethroids, e.g., deltamethrin and permethrin, may require the use of treated papers in tests much closer to time for treatment. Alternatively, if wanting to link actual dose with response, when repeating similar experimentation, the preferred method is to analyze the amount of AI present in the papers at the time of testing. The optimum time-lag between treatment and testing procedures needs to be explored further and verified for comparing test standardization and interpretation of findings with other compounds with high vapor pressure properties . Additionally, the physical/chemical effects of silicon oil (diluent) on the rate of vaporization needs clarification. Until there are a determination and agreement on preferred testing procedures, we provide a cautionary note when using treated papers in testing systems for the evaluation of TFT and similar compounds against insects.Ae. aegypti [Culex quinquefasciatus Say [An. minimus with increasing concentration when allowed direct contact with TFT, due to the stronger knockdown effect inside the chamber (non-escaping mosquitoes). A notable decrease in escape was also observed in Ae. aegypti between the three lower and highest concentrations, while An. dirus had the highest percent escape at the lowest concentration. The reduced escape is attributed to the stronger knockdown effect inside the chamber (non-escaping mosquitoes). In noncontact tests, colonized Ae. aegypti escape was greater at concentrations above the LC50. Although the repellency escape in DDC and TMMU anopheline strains tended to decline with increased concentration, all but one pairing (LC75 vs. LC99) found differences were not significant. Overall, both Anopheles species showed significantly stronger escape responses compared to Ae. aegypti (statistics not shown). Nentwig et al. [Ae. aegypti. Estrada et al. [Ae. aegypti and Ae. albopictus with increasing TFT concentrations ranging from 0.0005% to 10% using the high-throughput screening system.The degree of contact irritability exhibited by mosquitoes can vary according to species and the type and dose of insecticide used , 25. Hig aegypti  and Culeatus Say . Contrasg et al.  reportedAn. minimus and An. dirus. Contact excitation played a much larger role with colonized Ae. aegypti (USDA) with LC50, more comparable with repellency produced by LC75 and LC99, and a reduced excitation response for the DC. Conversely, using the LC50, the Ae. aegypti (NON) field population showed no contact response. Nentwig et al. [Ae. aegypti using three difference tests systems . Wagman et al. [The ER contact test design potentially measures the combined contact and noncontact mechansims of behavioral avoidance. Unlike the noncontact test design that measures spatial repellency only, the contact test cannot definitively separate between excitation and repellency responses. To estimate the effect of physical contact with a treated surface, mean contact escape responses were adjusted a second time see  using thg et al.  also shon et al.  also repAedes, Anopheles and Culex mosquitoes [The adjusted contact data showed colonized and field population anophelines produced relatively poor contact excitation compared to marked repellent activity. The robust repellency response seen with TFT is in contrast to other less volatile pyrethroids. Using the same ER assay system and a range of different pyrethroids, including deltamethrin, bifenthrin, permethrin, α-cypermethrin and λ-cyhalothrin, contact excitation appears the predominant mode of action for squitoes , 35, 36.squitoes .An. minimus was more susceptible to KD and mortality than An. dirus when exposed to 2% TFT for 60-min in a large semi-field enclosure . However, in this study, An. dirus showed much greater KD and mortality compared to An. minimus, while Ae. aegypti was less sensitive to KD (except for the DC). Exposing to TFT for 30-min or less (mosquito escaped from the treated chamber) may result in variable KD response for each mosquito species. Additionally, mosquitoes recovered within 24-h with decreased mortality in all treatment (except for Ae. agypti at LC50). In this study, spatial repellency of TFT occurred more quickly than a toxic response at test concentrations by causing mosquitoes to escape from the chamber. However, as shown in the recovery of the non-escape groups from knockdown, the test concentrations may not be high enough to completely kill all mosquitoes. As exposure time is one of the important criteria for evaluating the chemical action [Beside ER responses, mosquitoes encountering TFT can be knocked down (KD) or can be killed, depending on the exposure and species , 48. Marl action , furtherAn. minimus in the noncontact tests. This finding suggests that lower sublethal concentrations  TFT may be sufficient in producing strong spatial protection with low knockdown in ceratin mosquitoes . Grieco et al. [50 of deltamethrin, cyphenothrin, d-tetramethrin and tetramethrin were optimal concentrations for repelling Ae. aegypti. Therefore, this was the rationale for using the single TFT LC50 for field populations of Ae. aegypti and An. harrisoni in the ER assay system.Regardless of concentration tested, there was no significant difference in escape responses in o et al.  suggesteo et al. . Mongkalo et al.  also docAe. aegypti from Nonthaburi Province was resistant to TFT, while An. harrisoni from Kanchanaburi Province was fully susceptible at the discriminating concentration [Ae. aegypti NON population compared to the susceptible USDA strain. The USDA strain had a very low repellency response at LC50 compared to contact excitation, while the opposite was observed for the NON population. Similarly, Kongmee et al. [Ae. aegypti escaped the deltamethrin-treated ER chamber faster than a susceptible population. Conversely, Boonyuan et al. [Cx. quinquefasciatus escaped compared to four resistant field populations. Thanispong et al. [Ae. aegypti from various locations in Thailand had similar behavioral responses regardless of degree of background resistance to DDT and α-cypermethrin. It has also been suggested that if the chemical (i.e. TFT) can elicit the behavioral responses through spatial repellency and contact excitation actions, then the degree of resistance in certain mosquitoes to a toxic action would be minimal to no effect in disrupting human-mosquito contact [At time of testing, the field-collected ntration . Signifie et al.  found thn et al.  reportedg et al.  observed contact .An. minimus DDC laboratory strain and recent field An. harrisoni KAN population, genetically closely related ‘sibling’ species, are susceptible to TFT, a significantly  greater escape response was observed among DDC than KAN mosquitoes. Similarly, DDC had more rapid escape response to bifenthrin (pyrethroid) than recently field-collected An. minimus [An. harrisoni from same location (Pu Teuy Village) found weak behavioral responses to pyrethroids compared to An. minimus collected further north in Mae Sot District (Tak Province). Conversely, long-established laboratory strains of An. albimanus  [An. dirus  [Although the long-standing  minimus . Potikas minimus  using Anlle, FL)  and An. Bangkok)  showed rBangkok) , given oAn. arabiensis compared to houses in the unprotected village. They concluded diversion of An. arabiensis from repellent users to non-users. Therefore, it is important to be aware of possible diversion when implementing a spatial repellent as a transmission control measure. However, if applying a spatial repellent that able to protect multiple persons over wide areas , this could minimize the risk of possible diversionary effects [One possible outcome of using a spatial repellent is the diversion of repelled mosquitoes to other areas, thereby possibly increasing transmission risk elsewhere , 51. We  effects .Understanding the behavioral responses of mosquitoes to insecticides is relevant for supporting current mosquito control tools and development of novel ones. We encourage those investigating chemical properties for the control of mosquito-borne pathogens to include detailed observations on behavioral responses as well as toxicity. The use of varying concentrations of insecticide is preferred to observe the sublethal avoidance responses to each, especially so in spatial repellancy assays. In our view, even in the presence of significant physiological resistance, if an insecticide remains effective for controlling disease transmission by preventing human bite exposure via altered behavior it should continue to be part of the abatement arsenal. The ability to evoke strong mosquito avoidance behavior for reducing human-vector contact is invaluable for understanding a chemical’s properties to control transmission.vs. recent field collection) to TFT. Further studies on mosquitoes from different locations can help optimize currently available chemical-based intervention tools and spur the development of spatial repellency products using TFT and similar vapor-active compounds. Furthermore, both indoor and outdoor evaluations of TFT action protecting against mosquito bites using different delivery formats is required under natural settings.This study assessed adult female mosquito behavioral responses with exposure to TFT using an ER assay system. These findings demonstrate clear ER responses but also differences in avoidance behavior between mosquito species and conditions (colony S1 Table(*) significant difference at 95% CI; LC, lethal concentration; DC, discriminating concentration; NC; noncontact control, NT; noncontact treatment, CC; contact control, CT; contact treatment.(DOCX)Click here for additional data file.S2 TableLC, lethal concentration; DC, discriminating concentration.(DOCX)Click here for additional data file.


Cerasus humilis) is a perennial woody fruit  sequencing and high-throughput chromosome conformation capture (Hi-C) technologies  distribution, the estimated genome size of C. humilis is 228.20 Mb  were more abundant than DNA transposons (17.10%). Genome completeness was evaluated using the Benchmarking Universal Single-Copy Orthologs (BUSCO) method  and P. mume (Japanese apricot), and this clade showed a closer relationship with P. persica and P. dulcisi, which coincides with results observed in a previous study by Jiang et al. , P. mume (Japanese apricot), and P. armeniaca (Apricot), which are the closest species with a chromosome-level assembly. Firstly, the conservation synteny among the 8 pseudo-chromosomes of C. humilis was investigated. A total of 2,195 homologous synteny blocks were detected between pseudo-chromosomes, and some homologous blocks within pseudo-chromosomes were also observed . Total genomic DNA was extracted using a DNA Extraction Kit  following the manufacturer's protocols. The quality and quantity of total DNA were determined with 1% agarose gel electrophoresis and a NanoDrop 2000 spectrophotometer .The leaves of a 10-year-old The qualified DNA was randomly disrupted into 350 bp fragments and subjected to construct in the Illumina library using the standard protocol provided by Illumina . The paired-end sequencing was performed using the Illumina HiSeq 4000 system with paired-end 150 bp (PE150). After the quality control of raw data, a total of 20.86 Gb clean reads were generated and used for the estimation of genome size. We calculated the number of 17-mer from the clean reads using the Jellyfish version 2.0 , P. avium (L) L., P. mume (mei), P. dulcis Miller., Malus domestica Borkh., and Pyrus bretschneideri Rehder) were used to analyze gene families. An all-to-all BLASTP analysis of proteins with a length ≥50 amino acid (aa) was performed with an E-value of 10−5. The paralogous and orthologous genes were identified using OrthoFinder software . The conservation synteny among the eight pseudo-chromosomes of C. humilis was investigated. Meanwhile, the gene synteny among the genomes of C. humilis, P. mume, and P. persica were compared. Given that apricot is the closest species of C. humilis based on the result of the species tree, we compared the chromosome synteny of C. humilis and apricot.To investigate the chromosome evolution between SRR10913940SRR10913942. The sequences of genome assembly are available in the figshare with https://doi.org/10.6084/m9.figshare.11669673. The GFF file is deposit at the figshare with https://doi.org/10.6084/m9.figshare.11669514.The raw reads generated from Illumina platform, PacBio long reads and Hi-C data have been deposited in NCBI Sequence Read Archive (SRA) under the accession number SRR10912179, SRR10913200 and SRR10882935, respectively. The Illumina RNA173 seq data used for genome annotation was deposited in the NCBI SRA under the accession number: PW, XM, and JD conceived the study. SY performed bioinformatics analysis. PW and XM collected the samples and extracted the genomic DNA. XM, SY, and PW wrote the manuscript. JZ revised the manuscript. All authors read and approved the final manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.