Water scarcity is a world issue, and a solution to address it is the use of treated wastewater. Indeed, in these wastewaters, pollutants such as pharmaceuticals, pesticides, herbicides, and heavy ions can be present at high concentrations. Thus, several analytical techniques were initiated throughout recent years for the detection and quantification of pollutants in different types of water. Among them, the surface-enhanced Raman scattering (SERS) technique was examined due to its high sensitivity and its ability to provide details on the molecular structure. Herein, we summarize the most recent advances (2021–2023) on SERS sensors of pollutants in water treatment. In this context, we present the results obtained with the SERS sensors in terms of detection limits serving as assessment of SERS performances of these sensors for the detection of various pollutants. Nowadays, the decrease of the supply in water is a world issue due to the pollution of water. Moreover, various factors such as climate changes, population increase, and industrialization increase the demand in water ,2. Thus,In this mini-review, the goal is reporting on the most recent advances (2021–2023) in SERS sensors for the detection of compounds of interest (pollutants) in water treatment. Moreover, various review articles on the detection of pollutants for environmental analyses  are already available in the scientific literature, but with different focuses ,32,33,34SERS sensing is not an issue in water because the targeted molecules (here pollutants) may be detected within the water. Thus, the SERS technique is a perfect tool allowing a quick sensing of various molecules in water ,36. The Here, the detection of pollutants such as pesticides, herbicides, and organic dyes for water treatment is addressed see . We starConcerning to the detection of organic dyes, Li et al. have reported a LOD of nd water . For thind water . Furthernd water . LODs ofimproved . Anotherectively . The autl system . In anototspots) .block-poly(acrylic acid), called PS-b-PAA), which is decorated with silver nanoparticles [Moreover, Liu et al. studied the use of plasmonic sensors composed of silver nanocubes (AgNCs) having an elevated degree of purity for the detection of MG molecules in aquaculture water. Liu et al. assessed the LOD of MG molecules at re water . This LOre water . Chen eten AgNPs . To finiarticles   in river water . This SEolecules . Anotheras found . The SERobtained . Zhu et AgNCs/GO . Han et trometer . Thus, aarticles . To concd waters   in mineral and river waters with a promising SERS substrate . This SEubstrate . To finiubstrate  , and heavy ions in water treatment is introduced see . We begiRegarding the detection of pharmaceuticals and personal care products, Haung et al. investigated the sensing of the antibiotic quinoline in wastewater. The authors employed self-assembled gold nanoparticles with highly uniform gaps of 0.9 nm between Au nanoparticles producing various hotspots, thus inducing a good enhancement of the SERS effect. Therefore, a LOD of 5 stewater . An altestewater . In thisμg/L see d. This Lμg/L see a produciAnother strategy was developed by Berganza et al. for the detection of another antibiotic (ciprofloxacin = CIP) in water. This strategy consisted of the use of magnetite  coupled to an extraction process, both in a microfluidic chip . The priin water . Anotherin water . This costewater . Moreovestewater . To do tstewater . In anotstewater . Mao et stewater . Therewistewater . The intstewater . To finistewater .Concerning the detection of heavy ions, Zorlu et al. investigated the SERS detection of the heavy ion Cuap water . To do tobserved . With anin water . He et ain water . Anotheroworkers . This SE reached . In addiched see b,c. To dappeared .To conclude this part on the detection of heavy ions and, more generally, this mini-review, Logan et al. developed a technique of SERS detection of HgNSs-PEG) . The priseawater .To summarize, in this mini-review, we have described the most recent advances of SERS sensors of pollutants in water treatment. Due to the performances of these SERS sensors, limits of detection were achieved ranging from nductors  and organductors . Lastly,nductors ,88,89,90


Chen S and Wang ZP analyzed the data. Zhang WX and Zhou L contributed analysis tools. Chen S, Li SX, Wang ZP and Zhang WX provided critical inputs on design, analysis, and interpretation of the study. All the authors had access to the data. All authors read and approved the final manuscript as submitted.


Epiperipatus ohausi  is the first species known from Rio de Janeiro, and more than 120 years later a new species is described in the state of Rio de Janeiro (RJ). In this study, we describe the second species in the state of Rio de Janeiro, which we are naming in honor of the indigenous population called puri who resided in southeastern coastal Brazil. The species can be diagnosed mainly by large dorsal primary papillae close to the insertion of the legs drawing a light band from the anterior to the posterior region of the body, and large dorsal primary papillae alternating on the dorsal plicae. Moreover, they are recognized in vivo by the color of the diamond-shaped marks brownish orange on the dorsal portion of the body. Epiperipatus puri sp. nov. morphologically seems to be related to Epiperipatus acacioi  by the shape of the primary papillae apical piece and to E. ohausi by the resemblance of dorsal papillae. The phylogeny shows a close relationship between the new species and E. ohausi in a clade with a still undescribed species from Rio de Janeiro, Brazil located within the Atlantic Forest, one of the most threatened biomes in the world. The phylum Onychophora (velvet worms) has received more attention in the last decade than ever, especially the clade Neopatida , which resulted in the recent description of five species , designating Peripatus edwardsii Blanchard, 1847 as the type species and transferring more nine species to this subgenus . Since Epiperipatus is treated as genus, although this author did not explicitly elevate the rank from the subgenera of Peripatus. Peck cited it under an identification key for families and genera of Onychophora and made the combinations with the species without using Peripatus in the binomina , an extremely threatened biome , as part of the rebuilding of the burned MNRJ zoological collections. Epiperipatus puri sp. nov. is the first species of onychophorans deposited in MNRJ after its main building was lost in a ruthless fire. The collection of Onychophora was held together with the collections of Arachnida and Myriapoda, under the curatorship of Adriano B. Kury. Most of the three collections were lost in the fire except for material under loan and all the data, which were safe due to the routine backup policy of the curator  from Nova Iguaçu municipality, Rio de Janeiro, Brazil to detect diagnostic features of the new species: MNRJ 0056; 1♀; BRAZIL, Rio de Janeiro, Nova Iguaçu, Reserva Particular do Patrimônio Natural dos Petroleiros; 23.XII.2009; Costa, C.S., Giupponi, A.P.L. leg. MNRJ 0058; 1♂; same locality; 11.III.2010; Costa, C.S., Chagas-Jr, A., Giupponi, A.P.L., Kury, A.B. leg.The type-series is composed of six specimens deposited in Museu Nacional, Universidade Federal do Rio de Janeiro, Rio de Janeiro (MNRJ) (curator A. B. Kury) and Museu de Zoologia da Universidade de São Paulo, São Paulo (MZUSP) (curator R. Pinto-da-Rocha), both in Brazil. The specimens were collected between 2012 and 2018 in a small patch of tropical humid forest  Fig. 1)Fig. 1) aWe studied one of the specimens  using scanning electron microscopy (SEM) following in vivo were taken with a SONY Cybershot DSC-HX1 with built-in flash, or Canon EOS Rebel XS with a macro lens and flash circular camera. Images were edited using Adobe Photoshop CS5  will represent a published work according to the International Commission on Zoological Nomenclature (ICZN). This published work and the nomenclatural acts it contains have been registered in ZooBank, the online registration system for the ICZN. The ZooBank LSIDs (Life Science Identifiers) can be resolved, and the associated information viewed through any standard web browser by appending the LSID to the prefix Taxonomic resultsPeripatidae Evans, 1901Family Epiperipatus Clark, 1915Genus Epiperipatus purisp. nov.Epiperipatus [sp6]: Costa: 2016; Costa, Giribet and Pinto-da-Rocha: 2021: 6, 8, 25.–4Epiperipatus species with large dorsal primary papillae close to the insertion of the legs drawing a narrow light band from the anterior to posterior regions of the body. The distribution of large pale dorsal primary papillae alternates the dorsal plicae with clear differences in the number in two sequential plicae. Moreover, they are recognized in vivo by the color the diamond-shape marks brownish-orange (Diagnosis (based on a combination of characters as follows). h-orange –2D.Description of female holotype (MNRJ 0093).Measurements. Length 43; width 3.0, height 3.9.Color  each, absent in females. Anal glands are inconspicuous and represented only by two pores on the anterior margin of the anal aperture, absent in females.Type material. Holotype: MNRJ 0093, 1♀, BRAZIL, Rio de Janeiro, Cachoeiras de Macacu, Reserva Ecológica de Guapiaçu (REGUA), X.2012, A.P.L. Giupponi, J.S. Silva leg. Paratypes: MNRJ 0087, 1 unsexed specimen, MNRJ 0088, 1♂, same locality, 28.II – 02.III.2012, A.P.L. Giupponi, J.S. Silva leg; MNRJ 0107, 1 unsexed specimen, same locality, 19.III.2018, R.L.C. Baptista leg; MZUSP 0122, 1♀, same locality, 21.XII.2014, A. Ferreira, A.P.L. Giupponi, A. Rezende, C.S. Costa leg.Distribution. Only known from the type locality .Macro-Jê linguistic group. They inhabited, among other places, the mountain region of the Rio de Janeiro state where specimens of this species were collected. Noun in apposition.Etymology. The epithet puri (in apposition) refers to the Puri indigenous group belonging to the Remarks. Paratype. Length 12 to 22; width 1.0 to 2.5. Legs. 26  and 27  pairs of legs.Epiperipatus is one of the most speciose genera of Peripatidae  and a potentially undescribed species  and three other unnamed species from the state of Espírito Santo, Brazil, which is geographically close to the state of Rio the Janeiro and located within the Atlantic Forest.However recent studies based on the molecular data regards the genus as non-monophyletic . PhylogEpiperipatus purisp. nov. is characterized as a new species by the roundish insertion of dorsal papillae, the three posterior scale ranks, and two prolateral and one retrolateral foot papillae in the feet of the fourth and fifth oncopods , Epiperipatus tucupiEpiperipatus cratensisEpiperipatus diadenoproctusE. purisp. nov.oncopods . The preEpiperipatus paurognostusin vivo) and the fourth spinous pad can be complete or incomplete, in E. purisp. nov. the background color of the body is dark reddish-brown (in vivo) and the fourth spinous pad is complete. The apical piece is conical in Epiperipatus adenocryptus  by the shape of the apical piece of the primary papillae, however, E. purisp. nov. primary papillae are lighter than other papillae. E. ohausi and E. purisp. nov. bear dorsal papillae with similar shape and size, but the latter also bear accessory papillae on the flanks and uniform background color of body and oncopods.In ocryptus  and E. marajoara , and sph beckeri  and Epipitanicus , robust rbolicus , while tEpiperipatus are closer to Peripatus than to the remaining “Epiperipatus”, however, this could be confirmed only by the inclusion of Peripatus juliformis Guilding, 1826 (type species of Peripatus) in Peripatidae analyses, neither of both studies included this species due to the lack of specimens in the collections adequate for the analyses.Additionally, the results of Epiperipatus and Peripatus remain unclear, we cautiously preferred keeping the new species as Epiperipatus, as their putative closer species E. ohausi is currently classified. According to Peripatus due to the presence of only two pairs of pregenital oncopods with crural papillae, in Peripatus should be more than three. Also, according to Peripatus because it has three ranks of scales in the apical piece, Peripatus should have more ranks.Although the boundaries between Epiperipatus purisp. nov. is the 16th Epiperipatus species described from Brazil  . The resandscape .E. purisp. nov. is the first Onychophora to REGUA. This demonstrates the high value of this reserve for the recovery of the endangered area it is embedded.Although composed mainly of secondary forests, REGUA is important to the conservation of local fauna. The area is known for its rich fauna of birds , buttere.g., For more than 30 years, scientists advocated for the importance of invertebrates and their conservation, but the perspective has not changed much . The velE. ohausi  and Nova Iguaçu , with policies prioritizing the fight against poverty, environmental destruction, and historical deficit in science and education, Brazil is passing through severe economic, political, and social turmoil . RecentlAlthough fires in Amazonian Forest, Cerrado, and Pantanal are usually related to the replacement of natural vegetation by cattle ranching and soy crops, in the AF the deforestation is related to urbanization see  and presEpiperipatus purisp. nov. contributes to the knowledge of the biodiversity in a hotspot for conservation, the Atlantic Forest. We characterized the species morphologically with the use of SEMs and photographs, including in vivo (important for recognizing the species in the field). All the type material was collected in a private reserve that is contiguous to a State Protected Area, demonstrating the importance of this type of initiative. E. purisp. nov. was assigned to Epiperipatus such as the putative closer species, E. ohausi, but future studies could reveal the actual boundaries of the genera for there is molecular evidence that they could belong to Peripatus (Our description of eripatus . One oferipatus .10.7717/peerj.15384/supp-1Supplemental Information 1Click here for additional data file.


Zhang ZW, Li YL and Gao L performed the experiments. Ye GB and Ling B analyzed the data. Huang W and Ling B contributed analysis tools. Ye GB, Huang W and Ling B provided critical inputs on design, analysis and interpretation of the study. All the authors had access to the data. All authors read and approved the final manuscript as submitted.


Imported fire ants, an invasive species, are reported from multiple locales in Kentucky, USA from 2014 to 2022.Solenopsis invicta Buren (Red Imported Fire Ant), S. richteri Forel (Black Imported Fire Ant), and their hybrid form Solenopsis invicta X richteri have spread throughout portions of the USA, especially in the southeastern region. Imported fire ants are a serious invasive and economically significant species in the USA and elsewhere, and their spread into new parts of the country is of great concern. Although early models predicted that the fire ants would not be able to survive very far north into the USA, these ants have nonetheless successfully continued their spread into higher latitudes. Based on Cooperative Agricultural Pest Survey (CAPS) samples, the Mississippi Entomological Museum Invasive Insect Screening Center, at Mississippi State University, has verified the presence of imported fire ants collected in Kentucky at multiple locations from 2014 to 2022.Since their introduction into the United States in the early 1900′s, imported fire ants, namely  The Mississippi Entomological Museum Invasive Insect Screening Center, funded by the USDA Plant Protection Act Section 7721 (PPA7721), was established by the Mississippi Entomological Museum (MEM) at Mississippi State University to provide taxonomic services for Federal and state invasive insect survey programs in the eastern USA. These services were designed to speed up the identification and early detection of exotic species. Samples are received via in-person drop-off and mail services from participating state Cooperative Agricultural Pest Survey (CAPS) programs, PPA7721 Goal 1 Survey participants, and USDA Plant Protection and Quarantine programs in the USA. Upon arrival, samples are screened for target taxa and possible exotics are then examined by experts. Positive identifications are reported, and voucher specimens are pinned, labeled, and deposited in the Mississippi Entomological Museum (MEM) with data entered into the Symbiota Collections of Arthropods Network (SCAN). Additionally, representatives of native target taxa are also retained and curated into the museum collection. The deposition of accurately labeled and identified specimens into museum collections and reporting of findings are critical components of the monitoring process. Solenopsis invicta Buren (Red Imported Fire Ant), Solenopsis richteri Forel (Black Imported Fire Ant), and their hybrid form Solenopsis invicta X S. richteri, are native to South America and are seriously invasive and economically significant species in the USA and elsewhere. Here we report records of S. invicta, S. richteri, and S. invicta X S. richteri from Kentucky, provide information about identification, discuss their economic and medical importance, and give a brief history of their spread in the region. Imported fire ants (Hymenoptera: Formicidae) including Imported fire ants can easily be detected in the field by the large mounds they typically construct, their aggressive stinging behavior, relatively small size , and black and red coloration. Other diagnostic characteristics for the worker caste include the antennae having 10 antennomeres of which the apical two are enlarged forming a club, propodeum lacking adornment in the form of spines or other protuberances, waist with two nodes, and gaster with a prominent sting. Females are similar to workers, but larger  and have 11 antennomeres. Males are slightly larger than large workers , body concolorous black, head tiny in proportion to the body, eyes enlarged, and have 12 antennomeres [S. invicta from S. richteri may be difficult where their geographic ranges overlap because they create hybrids that have a mixture of features from the two species. Definitive identifications may require examination of cuticular hydrocarbons and venom alkaloids or DNA analysis [S. richteri  to new areas to start colonies , and queHuman activities play a significant role in the spread of invasive species including imported fire ants. Newly mated queens may be transported on motor vehicles on which they have landed. Colonies may be transported in the soil of nursery stock, mulch, sand, gravel, grass, sod, hay, wood, beehives, soil-moving equipment, and other inadvertent means ,11. S. richteri and S. invicta are thought to have first arrived via cargo ships in the USA in Mobile, Alabama with S. richteri arriving as early as 1918 and S. invicta likely in the early 1930s [S. invicta and S. richteri were not formally recognized as two distinct species until 1972, when Buren revised the group [S. invicta was introduced to the USA, it proved to be the more aggressive species and apparently displaced S. richteri from the Mobile area, with the idea being that S. richteri was continuously being forced east, north, and west by S. invicta. Trager [S. richteri “apparently occupied much of Mississippi and Alabama”, but specimen records do not reflect this. Instead, the movement of S. richteri may have been less linear, and instead patchier due to incidental movement by trains or other means. Historical records indicate that S. richteri (and possibly a hybrid form) was forced outward from the Mobile area into the southeastern coastal area of Mississippi and southwestern Alabama. By 1949, S. richteri was no longer found in those areas and was replaced by populations of S. invicta [S. richteri populations in those localities resulted from accidental introductions via trains. Although S. richteri was eliminated from the coast by fierce competition of S. invicta, conditions and isolation from S. invicta in these locations in central Alabama and Mississippi appeared to have allowed S. richteri to gain a stronger foothold. Eventually, S. invicta continued its spread and when it reached these northern populations of S. richteri, hybridization occurred between the two species at the edges of their distributions and pure populations of S. richteri were again pushed northward. This northward spread of S. richteri, stemming primarily from the disjunct central Mississippi populations, has continued north through western Tennessee and most recently into southwestern Kentucky, with S. invicta surrounding populations of S. richteri both to the south, west, and the east and the hybrid form forming a distinct “ring” between the two species. Solenopsis richteri has been reported in the literature from Arkansas [S. richteri was formerly recognized as a species and these unverified records seem unlikely. Examination of historical museum specimens from Arkansas by MacGown, as well as numerous collections by MEM staff and other researchers in Arkansas corroborate the idea that S. richteri has not been found in Arkansas. Both ly 1930s . It shouhe group , althoughe group  indicatehe group , after e. Trager  wrote th invicta . However invicta , Meridia invicta , Sessums invicta . These sArkansas ,16, but S. invicta would be limited to a latitude of approximately 42° along the Pacific coastline and approximately 37° elsewhere based on the cold tolerance of the ants. The authors did not estimate cold tolerance for S. richteri or S. invicta X richteri but speculated that they may have slightly more tolerance for cold temperatures than S. invicta. Overall, the model given by Korzukhin et al. [S. invicta would spread westward to California to the southern Oregon border and eastward up the Atlantic coast as far north as Virginia. However, imported fire ants have spread further north than predicted and have been found in areas once thought to have been improbable such as Oregon, Washington D.C., Maryland, and most recently Kentucky. Based on habitat and climate in their native ranges, a model of range expansion in the USA given by Korzukhin et al.  predicten et al.  has provS. invicta, S. richteri, and their hybrid from one another based on different cuticular hydrocarbon and venom alkaloid profiles as described in Menzel et al. [Specimens from possible fire ant mounds reported by Kentucky citizens were collected by CAPS employees and placed in ethanol and/or hexane and sent to the MEM Invasive Insect Screening Center for identification and verification. Identifications were made using morphological methods and the keys found in . When pol et al. . SpecimeThough the model given by Korzukhin et al.  predicteS. invicta. These specimens were not corroborated by the MEM. Three years later in 2012, fire ant mounds were again detected in Calloway County and confirmed by a USDA identifier as S. invicta [The earliest observations of imported fire ants in Kentucky were in October 2009 when mounds were detected during a CAPS survey at a nursery in Murray in Calloway County in the southwestern portion of the state . Mounds  invicta . S. richteri by a USDA identifier. The following year, a sample collected in Trigg County at the Lake Barkley Marina (36.8530–87.9422) on 14 October 2014 by J. Lensing was sent to the MEM for verification and specific identification. Specimens, which had been collected in ethanol, were identified as S. richteri based on morphological characteristics, and genetic assays agreed with this determination. In 2015, the MEM received additional samples of fire ants from Trigg County collected near the Golden Pond Visitor Center (36.7797–88.0670) on 19 May 2015 by J. Lensing. Samples were again collected in ethanol and based on morphological examinations and genetic assays the ants were determined to be S. richteri. Another sample was made in Trigg County near the Golden Pond Visitor Center (36.7797–88.0670) on 1 August 2015 by J. Lensing and sent to the MEM. Specimens were collected in both ethanol and hexane and again determined to be S. richteri. The hexane sample was very helpful as it allowed us to corroborate the S. richteri using gas chromatography and mass spectrometry methods. In July 2013, United States Forest Service personnel found fire ant mounds in the Land between the Lakes National Recreational Area in Trigg County, which is located in the southwestern part of the state. Specimens were collected and identified as S. invicta [S. invicta, which corroborated earlier identifications of fire ants from the same site. All mounds at the Kenlake sites were treated beginning in September 2017 and retreated in October 2017. The treatments appeared to be successful as no fire ants were found on return visits to the site after the collections were made in September 2017. Likewise, fire ants were not detected there during surveys in 2019 and 2021 [S. richteri, rather than S. invicta, which had been previously found at the site in 2017 and 2018. In 2017, 79 fire ant mounds were detected at the Kenlake State Resort Park in Marshall County, and specimens were identified as  invicta . On 18 SS. invicta X richteri.In 2022, the MEM received samples of fire ants collected by J. Collins from McCreary and Whitley Counties in the southeastern part of the state. Data for specimens were as follows: KY, McCreary Co., Strunk, 36.6409, −84.4100, 21 February 2022, J. Collins, collected at farm/residence; KY, McCreary Co., Stearns, 36.7164, −84.6099, 14 June 2022, J. Collins, collected at residence; KY, Whitley Co., Williamsburg, 36.6853, −84.2395, 22 September 2022, J. Collins, collected on roadside; KY, Whitley Co., Williamsburg, 36.7732, −84.0765, 22 September 2022, J. Collins, collected on farm; KY, Whitley Co., Williamsburg, 36.6409, −84.2589, 21 June 2022, R. Bessin, collected on farm. All samples from McCreary and Whitley Counties were morphologically identified as S. invicta and S. richteri in Tennessee, S. richteri populations were observed only in the western portion of the state from the northern border of Mississippi to near the southwestern border of Kentucky. According to Pandey et al. [S. invicta only comprised 2.3% of imported fire ants found during a survey done in Tennessee from 2004–2005, and colonies were only found in a few sites in the central and eastern parts of the state, mostly in metropolitan areas. Populations of the hybrid form were found to be widespread across the entire southern half of the state [Based on results from a recent paper by Pandey et al.  on the dy et al. , populathe state . S. richteri collected in Kentucky have only been found in the southwestern region directly north of a line following the distribution of this species from Mississippi through west and central Tennessee [Thus far, populations of ennessee . Similarennessee . S. invicta is thought to be less cold tolerant than S. richteri or their hybrid form [S. invicta reported from Tennessee were from Nashville, and that was the only locality in the entire western two-thirds of the state where it has been observed. It would not be a stretch to think that fertilized females of S. invicta might have been inadvertently transported from the Nashville area via truck along Interstate Highway 24 and Highway 80 to the site in Murray, Kentucky where this species was first found at a plant nursery. Due to S. invicta being less cold tolerant, it is probable that eradication efforts were successful. The discovery of S. invicta several years later in 2017 at the Kenlake State Resort Park was also likely due to anthropogenic movement of the species, as the area is known for outdoor recreational activities and fire ants could have easily been transported to the region. At first glance, it may appear surprising that both S. invicta and S. richteri were found in the same area at Kenlake Park, however, their occurrences there did not overlap temporally. Early populations of S. invicta were apparently eradicated, allowing S. richteri to successfully colonize the area later. Given that rid form , its preImported fire ants are clearly moving farther north than predicted, both naturally and by unintentional anthropogenic transport. As a result of the possible adaptation of the ants to colder temperatures and our climate becoming slightly warmer, imported fire ants are now likely established in Kentucky. Diligence when transporting various goods from quarantined areas is essential. Reporting to the Kentucky CAPS program of imported fire ant mounds is also critical in stemming their spread.


Lawsonia intracellularis, which infects the cells lining the ileum and infrequently cells in the large intestine, and it causes inflammation, hemorrhages, and thickening of the intestinal wall  prior to a combined L. intracellularis/PCV2 infection mitigated the clinical signs, lesions, and level of shedding of L. intracellularis infection. Prior PCV2 exposure also led to higher L. intracellularis antibody titers after the combined infection, which warrants further research into the cross-talk between the immune response to viral infections and L. intracellularis infection. The microbiota of the pig's ileum is known to influence the susceptibility to L. intracellularis, and the infection also adversely affects the gut microbiota . Particularly, an increase of pathobionts, including Collinsella, Campylobacter, Chlamydia, and Fusobacterium, was observed in the gut during L. intracellularis infection. Interestingly, the live attenuated L. intracellularis vaccine, in addition to decreasing the L. intracellularis shedding, significantly decreased the relative abundance of pathobionts, highlighting the role of the broader gut microbiome composition in the pathogenesis caused by L. intracellularis. Another article in this collection by Hankel et al., which is based on their analysis of fecal microbiota in pigs naturally infected with L. intracellularis that were varying in their vaccination status  and clinical conspicuity, found the influence of the microbiota on the severity of disease. The causal role of the microbiota in the pathogenesis of L. intracellularis infection is yet to be completely unraveled.Porcine proliferative enteropathy (PE) or proliferative hemorrhagic enteritis (PHE), or commonly “ileitis”, is a severe intestinal disease of domestic pigs and wild boars. It is caused by the bacterium nal wall . The disnal wall . L. intrhamsters . The etidinburgh . L. intrllularis . Now, thllularis , 5. The fections . A crossrom 2015 . The vaccrobiota . In thisL. intracellularis consists of a 1.46 Mb chromosome and three plasmids of 0.03, 0.04, and 0.19 Mb (NCBI Refseq IDs NC_020127 to NC_020130). A complete genome dataset of at least nine strains of L. intracellularis from around the world  are available in the public domain at the end of March 2023. Notably, the E40504 strain was isolated from a horse. The genome data have led to the development of several semi-quantitative and quantitative PCR assays for the diagnosis of L. intracellularis. Xiao et al. expressed the outer membrane protein (Omp2) of L. intracellularis from a synthetic gene based on sequence information. Monoclonal antibodies specific to Omp2 were produced and found to be efficient in detecting L. intracellularis in Western blots, immunohistochemistry, and immunofluorescence assays. Moreover, a comparison of the gene expression profiles of the pathogenic low-passage and non-pathogenic high-passage isolate of L. intracellularis revealed that there were only 401 genes expressed by the pathogenic bacteria , which is commonly found in pathogenic bacteria to secrete effector proteins into host cells. Through a yeast expression array expressing hypothetical genes from L. intracellularis, it has been identified that LI1035 interacts with the mitogen-activated protein kinase (MAPK) pathway and regulates actin organization in yeast . They found that L. intracellularis is endocytosed by both clathrin-dependent and -independent mechanisms, but the internalization of heat-killed bacteria is decreased in the clathrin-depleted cells. This suggests potential active mechanisms, involving a secretion apparatus, effected by L. intracellularis during endocytosis, as observed in many pathogenic intracellular bacteria. Given the wealth of knowledge on the genome of L. intracellularis, potential effector proteins or virulence factors remain to be characterized. With the current level of information on the biology of L. intracellularis, a promising outlook prevails over the efforts for a comprehensive understanding of the pathogenesis of this fastidious organism, which may require uncharacterized inciting factors to cause disease.The reference genome of rea CBNU0, and Chiammation . The stiin yeast . PereiraThe author confirms being the sole contributor of this work and has approved it for publication.


Diplolepis ogawai Abe and Ide sp. nov. (Hymenoptera: Cynipidae) induces galls on Rosa hirtula (Regel) Nakai , which is endemic to a restricted area of Honshu, the main island of Japan. The gall is induced mainly on the leaf of R. hirtula in spring and the mature gall falls to the ground in early summer. The gall-inducing wasp emerges from the gall on the ground in the following spring, suggesting that D. ogawai is univoltine. From spring to summer, the braconid Syntomernus flavus Samartsev and Ku and the eulophid Aprostocetus sp. are parasitic on the larva of D. ogawai in the gall, and the adult wasp of both parasitoid species emerges from the gall on the ground in summer. For S. flavus, this is the first distribution record in Japan and the first host record. Since R. hirtula is threatened with extinction by succession and deforestation, D. ogawai and its two parasitoid wasp species are considered to be at risk of coextinction with the threatened rose. In the event that the population size of this rose species is further reduced, D. ogawai and its parasitoids may become extinct prior to the extinction of R. hirtula. To conserve these three wasp species associated with R. hirtula, protection of remnant vegetation where individuals of this threatened rose species grow is necessary. Rosa hirtula (Regel) Nakai  is endemic to a limited area of Kanagawa, Shizuoka, and Yamanashi Prefectures on the main island of Honshu, Japan , which consist of the genera osa spp. . The gloer clade , 2020. Her clade . Based oin China , the exiin China . In recescovered .R. hirtula shrubs. He observed that gall wasps and their parasitoids emerged from the galls he reared. Preliminarily, morphological examinations of these wasps revealed that the gall wasp was a member of Diplolepis, and the parasitoids consisted of one braconid and one eulophid species. Many rare and undescribed species of insects become extinct before description  fitted with digital single-lens reflex camera , and under a scanning electron microscope  at 1.5 kV. The length of body parts was measured with an ocular micrometer. Focus stacking was performed using CombineZP software (available from: https://combinezp.software.informer.com/) for the microscopic images. All images were processed and assembled using the GNU Image Manipulation Program .In 2020 and 2021, fresh galls that had fallen to the ground from uo Ogawa . After kD. ogawai, the following morphological abbreviations are used: POL, postocellar line ; OOL, ocular–ocellar line ; and LOL, lateral–ocellar line . The morphological terminology for gall wasps follows In the description of D. ogawai and its parasitoids emerged was observed and their diameters were measured with digital calipers to clarify the effects of parasitism on gall shape and size. The galls were then dissected under a binocular stereomicroscope to determine their contents. To compare the structure of the galls on R. hirtula by D. ogawai with that of the galls induced by Diplolepis japonica , galls of the latter species were collected from leaves of Rosa multiflora Thunb. by Yoriko Abe in Kyonan-cho, Musashino City, Tokyo on 25 May 2021 and were reared under field conditions on the Ito Campus of Kyushu University until 26 March 2022 when the female wasps emerged. These wasps were identified as D. japonica based on The shape of nine galls from which wasps of D. ogawai and that of the braconid using previously described methods (https://www.ncbi.nlm.nih.gov/genbank/) under the accession numbers OP281689 for D. ogawai, OP281690 for the braconid and OP281691, OP281692 for the eulophid. The COI sequence of D. ogawai was compared with those of other congeners in GenBank using BLAST .Partial sequences (658 bp) of the COI gene of mtDNA were determined for one wasp of  methods . DNA of  methods . PCR conThe data of all the sample details are provided in www.zoobank.org), the official register of the International Commission on Zoological Nomenclature. The LSID (Life Science Identifier) number of the publication is: urn:lsid:zoobank.org:pub:89AAB7FF-3A3B-49B1-9493-D67A0FB72B4A.This paper and the nomenclatural act(s) it contains have been registered in Zoobank (D. ogawai and other congeners were detected and this new species is described below. The parasitoids reared from the new Diplolepis were identified as Syntomernus flavus Samartsev and Ku (Hymenoptera: Braconidae) and Aprostocetus sp. (Hymenoptera: Eulophidae) based on Morphological differences between (urn:lsid:zoobank.org:act:24F3E6F2-081B-4A3A-A518-FDF2157E7785)Rosa hirtula-leaf-gall wasp in Japanese[Japanese name: Sanshôbara-ha-tamabachi], which means  and 2See Body  black. MHead  and C seMesosoma  dorsallyWing surface and margin closely ciliated. Marginal cell of forewing closed with pale anterior margin, 2.3 times as long as wide, 2r curved without median prolongation into the marginal cell. Areolet indistinct. Hind femur without flange. Apex of metatarsal claw bent; base not expanded to lobe. Lengths of forewing and hind tibia 2.75 and 0.96 mm, respectively.Metasoma  smooth, Not known.n = 4) 2.75–3.11 mm, and of hind tibia 0.96–1.03 mm.Areolet indistinct or absent. Lengths of forewing  metasoma is dark brown and partially reddish-brown in D. japonica, but black in D. ogawai; 2) mesoscutal impression is present in D. japonica, but absent in D. ogawai; 3) areolet is distinct in D. japonica, but indistinct or absent in D. ogawai; 4) forewing is infuscate along all the veins of the marginal cell in D. japonica, but not infuscate in D. ogawai  metasoma is chestnut brown in D. valtonyci, but black in D. ogawai; 2) mesoscutal impression is present in D. valtonyci, but absent in D. ogawai; 3) forewing is infuscate along all the veins of the marginal cell in D. valtonyci, but not infuscate in D. ogawai.The new species is named in honor of Haruo Ogawa, who collected the specimens.D. ogawai can be characterized as follows. Galls are induced on both the upper and lower surfaces of the R. hirtula leaf . In contrast, the gall induced by D. japonica bears sharp-pointed spines . The gall wall is thin (ca. 0.2 mm) in D. ogawai, but thick (2.0–2.7 mm) in D. japonica  Nakai (family: Rosaceae)Japan (Kanagawa and Shizuoka Prefectures in Honshu).A univoltine life cycle was indicated by rearing the wasps .See Diplolepis ogawai Abe and IdeSouth Korea , Japan  of S. flavus from summer to the following spring is unknown.The members of See Diplolepis ogawai Abe and Ide.Japan (Kanagawa and Shizuoka Prefectures in Honshu).Aprostocetus contains more than 800 species worldwide , a Japanese species, has not been amended since its original description  of Aprostocetus sp. from summer to the following spring is unknown.As in Diplolepis radicum (Osten Sacken), Diplolepis rosae (L.), and Diplolepis fusiformans (Ashmead) as having the most similar COI sequences to D. ogawai, with COI sequence homologies in the range 89–90% compared to D. ogawai.The BLAST analysis identified Baryscapus pallidae Graham and Aprostocetus cerricola (Erdös) as the two species with COI sequences most similar to Aprostocetus sp. These two species had COI sequence homologies in the range 90–92% compared to Aprostocetus sp.In addition, the BLAST analysis selected D. ogawai. In addition, two parasitoids, S. flavus and Aprostocetus sp., were reared from D. ogawai. This is a new distributional record for S. flavus from Japan.In this study, we described the gall wasp, D. ogawai and its parasitoid Aprostocetus sp. result in about 90% match. Moreover, the three best hits for D. ogawai are two Nearctic and one European species, and the two best hits for Aprostocetus sp. are European species. Such a situation is ascribable to the lack of appropriate sequence data from most of the Asian representatives of Diplolepis spp. and their parasitoids. Effective DNA barcoding requires a well-sampled, extensive reference database. Reliable DNA-based determination of phylogenetic position of this new species needs nuclear data as well as better Asian sampling.Both BLAST searches for Diplolepis induce galls on plants of the genus Rosa  in Rosa pimpinellifolia L., no reliable data indicates that Diplolepis species are host-specific in Europe at the level of host plant species (D. japonica has been recorded from R. multiflora and R. rugosa (D. japonica is widely distributed in East Asia, including Japan, Korea, and China (D. japonica could be an undescribed species. In contrast to the typical Diplolepis species, galls of D. ogawai have been only recorded from R. hirtula. The exceptionally high host specificity of D. ogawai to R. hirtula, which has a very limited natural distribution, most likely means that the distribution range of this gall wasp is restricted to an area on the main island of Japan. The vulnerable rose R. hirtula is considered to be a keystone species within a local ecological community (a network in which species are trophically connected; e.g., R. hirtula to the newly discovered fauna associated with it.Because the members of sa e.g., , the disg., Rosa . However species . In the . rugosa . Since t. rugosa , D. japond China . HoweverD. ogawai means that it likely does not adversely affect the host plant population. Therefore, when R. hirtula suffers anthropogenic habitat degradation, the extinction of D. ogawai due to the decline of the host plant populations is rather likely to come earlier. R. hirtula is distributed in a limited area (R. hirtula was discovered. The new gall wasp D. ogawai appears to be host-limited to the rare rose, and two parasitoids: S. flavus and Aprostocetus sp. depend on this gall wasp from spring to summer. These three wasp species are also considered to be at risk of coextinction with this vulnerable plant as follows: the primary extinction of R. hirtula causes the secondary extinction of D. ogawai, followed by the tertiary extinction of S. flavus and Aprostocetus sp. Generally, host-limited herbivorous insects may become extinct before their host plants (R. hirtula is reduced further, these three wasp species may become extinct before the vulnerable host plant. Therefore, the insect fauna depending on the rare rose is considered vulnerable. As pointed out by Although some species of cynipid gall inducers often cause serious damage to their host plants , the rarted area  and is tted area . Moreoveted area . In the t plants . If the


High-speed running and sprinting in professional adult soccer: Current thresholds definition, match demands and training strategies. A systematic review By Gualtieri A, Rampinini E, Dello Iacono A and Beato M. (2023) Front. Sports Act. Living 5:1116293. doi: 10.3389/fspor.2023.1116293A Corrigendum on Additional Affiliation(s)In the published article, there was an error regarding the affiliation for Ermanno Rampinini. As well as having affiliation 3, he should also have:Sport and Exercise Discipline Group, Human Performance Research Centre, Faculty of Health, University of Technology Sydney, Moore Park, New South Wales, Australia.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.


Pseudopungtungia tenuicorpa. This species was found to preferentially select cracks as its spawning habitat, followed by the nests of other fish. In particular, cracks smaller than 5 cm provided a safe area for the spawned eggs while being difficult for other fish to invade. P. tenuicorpa eggs spawned in cracks larger than 6 cm were continuously consumed or damaged by other predatory fish. In South Korea, cracks smaller than 5 cm are more abundant in the upstream parts of the Han River than in other river basins (Geum River and Nakdong River), hence explaining the limited spatial distribution of P. tenuicorpa.This study explained the effects of the type and abundance of micro-topography in a river basin on the geographical distribution of the endemic fish Pseudopungtungia tenuicorpa in South Korea is closely related to the local abundance of their preferred spawning habitats. Eggs of P. tenuicorpa were most frequently observed in cracks smaller than 5 cm, followed by the nests of other fish, such as Coreoperca herzi. Although eggs of P. tenuicorpa were also found in cracks larger than 6 cm, most of them were consumed or damaged by other fish. However, the number of P. tenuicorpa eggs spawned in cracks smaller than 5 cm remained similar to the number of eggs at the time of initial spawning, even after 9 days. The strong preference of P. tenuicorpa for cracks smaller than 5 cm was found to influence the geographical distribution of P. tenuicorpa in South Korea. In the upper reaches of the Han River, the ratio of cracks smaller than 5 cm is higher than that in the Geum River and in the Nakdong River, which contributes to the stable population growth of P. tenuicorpa there. In the future, management strategies to maintain/restore cracks smaller than 5 cm should be reviewed for the preservation of this endemic fish in South Korea.In this study, we hypothesized that the limited geographical distribution of  The selection of an efficient spatial location for spawning is important for securing the stable population growth of freshwater fish by increasing the hatching rate of spawned eggs ,2. SincePseudobagrus sp.), Coreoperca herzi, and bass  cope with predators or anthropogenic disturbances (such as oxygen deficiency) by protecting their nests during spawning [The construction and selection of efficient nesting locations are coordinated based on local environmental conditions and interaction between individuals or communities ,8. Changspawning ,15.Pungitius sinensis consume a large amount of energy to protect their spawned eggs after nest formation and die soon after the birth of their young ones [However, although the behavioral strategies of freshwater fish for selecting spawning sites or for protecting their nests are suitable for increasing the hatching rate, they are associated with high energy consumption and high costs . For exaung ones . While eRhynchocypris kumgangensis and Hemibarbus mylodon are distributed only in the upper-middle reaches of the Geum River and the Han River, located in the central part of South Korea, and Pseudobagrus brevicorpus, Koreocobitis naktongensis, and Microphysogobio rapidus are distributed only in the Nakdong River basin, located in the southeastern part of South Korea [Coreoperca kawamebari and Acheilognathus somjinensis are only observed in the Yeongsan River basin, located in the southwestern part of South Korea [Acheilognathus lanceolatus, Rhodeus sericeus, and Acheilognathus rhombeus have a habit of spawning on the bodies of freshwater clams [Synodontis multipunctatus) is mainly distributed in Lake Tanganyika, in which cichlids are abundant, since they are known to breed via brood parasitism in the mouth-brooding cichlids [Selecting a locally abundant space as a spawning site is an efficient strategy that can reduce the energy cost toward the search for appropriate nest . Obviousth Korea ,21. Coreth Korea ,21. Moreer clams , and hener clams . Similarcichlids ,25.Pseudopungtungia tenuicorpa with clear geographical distribution limits. Pseudopungtungia tenuicorpa is a native Korean fish species belonging to the genus Pseudopungtungia, which was recorded as a new species in 1980, and is only observed in some upper parts of the Han River. This area not only has continuous water flow but is also covered with giant rocks and gravel, supported by the typical environmental characteristics of the upper reaches of the river [Pseudopungtungia nigra, which belongs to the same genus as P. tenuicorpa, is distributed in the upstream area of the Geum River, which is clearly distinguished from the distribution area of P. tenuicorpa; the two species do not coexist in one area. While P. nigra is known to spawn via brood parasitism in the nests of Coreoperca herzi, distributed in the same area [P. tenuicorpa is still insufficient. Since C. herzi is distributed not only in the Geum River basin but also in most river basins of South Korea, including the Han River, where P. tenuicorpa is observed, we assumed that P. tenuicorpa is also spawned in the nests of C. herzi, just like P. nigra. However, based on the fact that P. tenuicorpa is not observed in river basins other than the upper part of the Han River, the topographical structure unique to the upstream section of the Han River may possibly be used as a spawning habitat. In addition, it is possible that the difference in geographical distribution between river basins has an influence on the selection and behavior of the two fish species (P. tenuicorpa and P. nigra), belonging to the genus Pseudopungtungia, with respect to their different spawning habitats.In this study, we focused on the spawning habitat selection and behavioral strategies of he river . Pseudopame area , researcP. tenuicorpa across potential spawning habitats, such as cracks, the nests of C. herzi, and the nests of other fish; (2) determine the spawning rate, egg number, and egg preservation rate of P. tenuicorpa based on the size class  of the crack; and (3) determine the brood parasitism rate of P. tenuicorpa in the nests of C. herzi. We hypothesized that the unique topographical characteristics of the upper reaches of the Han River would lead to differences in the behavior and spawning habitat selection of P. tenuicorpa as compared to those of P. nigra in the Geum River. In this study, five streams  were selected based on the distribution area of P. tenuicorpa, as recorded in previous studies [P. tenuicorpa, the rate of choice of an alternative spawning habitat over the preferred habitat was expected to increase if there was a difference in the abundance of spawning habitats in each river surveyed. The results of this study can contribute to the identification of geographical distribution limits within which P. tenuicorpa is distributed and will help to establish management strategies for the preservation of P. tenuicorpa in the future.In this study, we aimed to (1) identify the egg observation frequency (%) of  studies ,29,30. D2). The Han River is largely formed of the Namhan River and the Bukhan River entering from the south and north, respectively, and then exiting into the West Sea of South Korea. This is in contrast to the section that passes through the downstream area after the confluence of the Namhan River and the Bukhan River. The upper reaches of the Han River are home to native Korean fish species, such as Brachymystax lenok, Hemibarbus mylodon, Rhynchocypris kumgangensis, Rhynchocypris kumgangensis, and Phoxinus phoxinus [The Han River is located in the northern part of South Korea and has the second longest basin length (481.7 km) after the Nakdong River (510.36 km). However, comparing the basin areas, it has the largest basin , a determination of the environmental variables, exploration of the spawning habitat, and investigation of the spawning behavior of P. tenuicorpa were conducted. P. tenuicorpa were frequently observed in the previous studies [During the spawning period of  studies ,29,30, t studies .P. tenuicorpa was collected from five points. The collection process was conducted using cast nets (7 × 7 mm) and scoop nets (5 × 5 mm) along 500 m transects at intervals of 1 km per section, similar to the measurement points of the environmental variables. The cast and scoop nets were deployed for 30 min and 15 min, respectively. Each of the collected fish was identified up to the species level, according to Kim and Park [After measuring the environmental variables, and Park , based oand Park .P. tenuicorpa in each stream section (5 km), snorkeling was used during the spawning period (April to June) to scrutinize the eggs of P. tenuicorpa spawned in cracks, the nests of other fish such as C. herzi, the surfaces of aquatic plants, and large rocks or gravel surfaces. The results were classified into four types of spawning habitats based on the spawning ratio of P. tenuicorpa eggs in each spawning habitat: (1) cracks, (2) the nests of C. herzi, (3) nests of other fish, and (4) others (plant surfaces and rock/gravel surfaces). The egg observation frequency (%) of P. tenuicorpa in the four types of spawning habitats was calculated as follows:In order to explore the spawning habitat of P. tenuicorpa, cracks were mostly found in large rocks . In each stream, the proportion of gravel less than 50 cm was relatively low, and most of the bottom layers were supported by giant rocks, making it difficult to consider cracks between pieces of gravel.As spawning habitats for C. herzi were also found under or on the sides of large rocks. The nests were present as a single clutch, but it was difficult to count the number of clutches in which P. tenuicorpa or P. herzi had spawned in these spaces.At the study sites, the nests of P. tenuicorpa, additional investigations were conducted on the spawning characteristics and behavior in cracks, where the eggs of P. tenuicorpa were most frequently found. First, snorkeling was used to measure the size of the cracks where P. tenuicorpa eggs were spawned or likely to be spawned, and accordingly, they were divided into three classes . We explored all the existing cracks within each stream section (5 km) for 5 to 6 days and attempted to find all the possible cracks. Then, the number of cracks, the spawning rate (%) of P. tenuicorpa, and the number of eggs were measured for each size class. In cases where a crack was too small or it was difficult to observe the eggs with the naked eye, an underwater piping endoscope camera  was used for investigation. The spawning rate of P. tenuicorpa, according to the size class of each crack, was calculated using the following calculation formula:Among the four spawning habitats of P. tenuicorpa were randomly selected for each size class , and the number of P. tenuicorpa eggs was measured over 9 days for a total of 15 cracks. The 15 cracks were selected as the points where eggs were spawned by P. tenuicorpa less than a day later. After marking the selected 15 crack entrances with PVC bars, the number of eggs was counted through snorkeling during the morning (8:00 to 10:00 A.M.). A total of five  measurements were made on alternate days to eliminate the effect of disturbance  on the measurement of the number of eggs in each crack as much as possible. The preservation rate of the eggs in each crack was calculated as follows:In order to determine the egg preservation rate (%) over time in each crack, 5 cracks spawned by P. tenuicorpa and P. herzi. The sharing status of spawning habitats was divided into four types . The observation frequencies (%) were calculated according to the three crack size classes .An additional investigation was conducted to determine whether spawning habitats (such as cracks) were shared between fish species, such as P. tenuicorpa in the nests of C. herzi in each stream section (5 km) was estimated after an investigation of the spawning behavior of P. tenuicorpa in the cracks. To determine the brood parasitism rate (%) of P. tenuicorpa in the nests of C. herzi, we used snorkeling to find all the nests of C. herzi located in each stream section, from which the number of C. herzi nests spawned by P. tenuicorpa was counted. The number of C. herzi spawned by P. herzi also helped to determine the sharing status of the spawning habitats based on the nests of C. herzi spawned by both P. tenuicorpa and P. herzi in cracks. Similar to the considerations for cracks, the sharing status of the spawning habitats was divided into four types . From these findings, the brood parasitism rate (%) of P. tenuicorpa and P. herzi in the nests of C. herzi was calculated as follows:The brood parasitism of C. herzi, 5 nests spawned by P. tenuicorpa were randomly selected, and the number of P. tenuicorpa eggs was measured over 9 days for a total of 15 nests. These 15 nests were selected considering those spawned by P. tenuicorpa less than a day later. After marking the selected 15 nest entrances with PVC bars, the number of eggs was counted through snorkeling during the morning hours (8:00 to 10:00 A.M.). A total of five  measurements were made on alternate days to eliminate the effect of disturbance on the measurement of the number of eggs in each nest as much as possible. The preservation rate of the eggs in each nest was calculated as follows:In order to determine the egg preservation rate (%) over time in the nests of C. herzi spawned by P. tenuicorpa per stream section were investigated. Underwater cameras were installed to observe the invading and spawning times of P. tenuicorpa in each nest of C. herzi for approximately 8 days during the spawning period of C. herzi; the invading and spawning rates of P. tenuicorpa per day were determined using the recorded data. The invading (6) and spawning rates (7) were calculated as follows:Furthermore, 10 randomly selected nests of C. herzi identified in the Jucheon Stream and Pyongchang Stream, located in the Han River area, were compared to those in the two streams of the Geum River (upstream of Geum River and Gurang Stream) and the three streams of the Nakdong River . From each stream, two sections (5 km) were selected, as in the case of the Han River, and the numbers of cracks and nests of C. herzi located in each section were counted through snorkeling. The cracks were divided into <5 cm, 6–10 cm, and >11 cm sections to maintain consistency in the investigation.The numbers of cracks and nests of P. tenuicorpa and the ten environmental variables. Furthermore, one-way analysis of variance (ANOVA) was used to examine the differences in (1) the ten environmental variables between the streams; (2) the number of cracks, spawning ratio, and egg number in the cracks; and (3) the numbers of cracks per size  and nests of C. herzi between the seven streams . Tukey’s honest significant difference (HSD) test was performed for additional post hoc comparison analysis in order to determine which of the differences were statistically significant. This statistical analysis method is suitable for identifying differences between values and has been used in various ecological studies. When applying statistical analysis, we used log-transformation to convert the data . This measure minimized the differences in the range of values so that statistical differences could easily be identified. The statistical analyses were performed using SPSS ver. 20 . Differences and relationships were considered significant at p < 0.05.We utilized a stepwise multiple regression analysis to examine the relationship between the abundance of P. tenuicorpa. In this area, the water velocity was in the range of 1.8 to 2.2 m/s, and the DO was in the range of 124 to 131%, which is characteristic of the upper part of the stream. The conductivity (134–164 µs/cm), turbidity (2.2–2.8 NTU), TN (0.613–0.684 mg/L), TP (0.011–0.016 mg/L), and Chl.a (0.8–1.3 µg/L) were found to have relatively low values and supported by the good water quality. The ten environmental variables no significant differences between the five streams (p > 0.05). In contrast to the environmental variables, the density of P. tenuicorpa was clearly different across the five streams , p < 0.05). The comparison using post hoc tests revealed significant differences between three groups  and Pyeongchang Stream ; B-group = Gapyeong Stream ; and C-group = Inbuk Stream  and Soyang Stream ). P. tenuicorpa was the most abundant in Jucheon Stream and Pyeongchang Stream, whereas it was lowest in Gapyeong Stream. The results of multiple regression analysis between the abundance of P. tenuicorpa and ten environmental variables were not statistically significant (p > 0.05).P. tenuicorpa were most frequently observed in cracks, followed by the nests of C. herzi  of P. tenuicorpa from Jucheon Stream and Pyeongchang Stream was more than 80%, and that in the remaining three streams was in the range of 40–60%. The spawning rate of P. tenuicorpa in 6–10 cm cracks in Gapyeong Stream, Inbuk Stream, and Soyang Stream was in the range of 41–44%, whereas that in Jucheon Stream and Pyeongchang Stream was less than 20%. The results of the post hoc tests showed significant differences in spawning ratio (%) between the three size classes of cracks in Jucheon Stream and Pyeongchang Stream , B-Group = 6–10 cm , and C-group = >11 cm ; Pyeongchang Stream, A-group = <5 cm , B-Group = 6–10 cm , and C-group = >11 cm ).The spawning rate (%) of P. tenuicorpa were spawned in cracks larger than 11 cm in Gapyeong Stream. The number of P. tenuicorpa eggs in each crack ranged from 800 to 1000, and the difference based on crack size was minimal. In addition, there was little difference in the number of P. tenuicorpa eggs between the streams.No eggs of P. tenuicorpa spawned in different cracks gradually decreased over time (The egg preservation rate (%) of ver time . HoweverP. tenuicorpa or P. herzi alone. The fish mainly spawned in cracks larger than 6 cm and preferred cracks larger than 11 cm as spawning habitats. Cracks smaller than 5 cm were spawning habitats only for P. tenuicorpa, and eggs of P. herzi were not found in them. The eggs of P. tenuicorpa were found in cracks of all size classes, being the most abundant in cracks smaller than 5 cm.No cracks of any size class were shared as a common spawning habitat among more than two fish species . Each crP. tenuicorpa in the nests of C. herzi was different between the five streams (p < 0.05). The brood parasitism rate (%) of P. tenuicorpa from Jucheon Stream and Pyeongchang Stream was low (less than 20%), whereas it was relatively high (at 64%) in Gapyeong Stream. In Inbuk Stream and Soyang Stream, it was 38% and 39%, respectively. Moreover, the nests of C. herzi were not shared with other fish, similar to the cracks. In most of the C. herzi nests, the eggs of P. herzi were observed more frequently than those of P. tenuicorpa. As an exception, in the nests of C. herzi in Gapyeong Stream, the egg observation rate of P. tenuicorpa was higher than that of P. herzi. In contrast to the case of fine stone meat, the brood parasitism rate of P. herzi in the nests of C. herzi was similar, in the range of 28–33% for all streams. However, except for Gapyeong Stream, the proportion of unspoiled nests of C. herzi spawned by P. tenuicorpa or P. herzi was higher than that of the nests of C. herzi in the four remaining streams.In each stream section, the brood parasitism (%) of P. tenuicorpa spawned in the nests of C. herzi in each stream were stably preserved for 9 days , excluding Gapyeong Stream, the early invading and spawning rate of P. tenuicorpa was less than 35%, but in Gapyeong Stream, the rates were 62% and 58%, respectively. Six to seven days after the early invasion and spawning of P. tenuicorpa in all the streams, no further invasion and spawning of P. tenuicorpa occurred in the nests of C. herzi.The early invading and spawning rate in the nests of  streams . The invP. tenuicorpa in the seven streams, including two streams of the Han River, the numbers of cracks were found to differ between the streams (Comparing the abundance of cracks which were spawning habitats for  streams .p < 0.05), whereas the number of cracks larger than 21 cm was similar between the streams (range 23 to 29). Cracks smaller than 10 cm and 11–21 cm were the most abundant in Jucheon Stream and Pyeongchang Stream, located in the Han River basin , whereas the remaining streams  located in the Geum River and Nakdong River areas were characterized by relatively small numbers of cracks. In contrast, the number of C. herzi nests was barely different between the streams , ranging from 12 to 15.Among the three ranges of crack sizes, the numbers of cracks of two size classes (>5 cm and 6–10 cm) were clearly different between the seven streams , excepting Jucheon Stream and Pyeongchang Stream, the ratio of selection of a crack smaller than 5 cm and of a gap of 6–10 cm as a spawning habitat was relatively high. This implied an increase in the density of P. tenuicorpa in cracks of 6–10 cm as an alternative habitat due to the lower availability of cracks smaller than 5 cm in the three abovementioned streams.In this study, P. tenuicorpa for cracks smaller than 5 cm can be understood from the results for the egg preservation rate of each crack size class. Eggs scattered in cracks smaller than 5 cm had a relatively low decreasing rate even after 9 days, whereas only approximately 25% of the eggs were preserved in cracks measuring 6–10 cm and >11 cm after 9 days. This meant that cracks of 6–10 cm and >11 cm, as spawning habitats for P. tenuicorpa, were riskier than those smaller than 5 cm. We frequently observed predators, such as Pseudobagrus sp. or Odontobutis platycephala, in cracks of around 6–10 cm and >11 cm, even during our investigations of the egg retention rates in these gaps. Therefore, the abundance of P. tenuicorpa in the upper reaches of the Han River can be considered to be mostly due to individuals hatched from eggs spawned in cracks smaller than 5 cm.The strong preference of P. tenuicorpa in the nests of C. herzi in the upper reaches of the Han River is due to the presence of an optimal spawning habitat, namely, the ‘crack’, in this area. Using the crack as a spawning habitat has the advantage of investing a relatively low cost while efficiently avoiding predators. In order for P. tenuicorpa eggs to be spawned in a nest of C. herzi, this not only requires the invasion of the nest of C. herzi but will also cost energy in order to resist the defensive behavior of the male protecting the nest. Although the eggs of P. tenuicorpa that are successfully scattered in the nests of C. herzi are protected by C. herzi males so that they are not consumed by other fish, the disadvantages of high energy consumption and hatching/spawning failure during the brood parasitism process include high energy loss. However, the fact that a nest of C. herzi is selected less frequently by P. tenuicorpa does not mean that this is a spawning habitat that they avoid. As described above in the previous paragraph, the selection of a spawning habitat by P. tenuicorpa is flexible, depending on the available environmental conditions. In streams where cracks were abundant, P. tenuicorpa had a high rate of selection of cracks as spawning habitats, even if there were abundant nests of C. herzi, but in streams (Gapyeong Stream) where there were relatively fewer cracks, the proportion of P. tenuicorpa selecting the nests of C. herzi increased. This implied that the nest of C. herzi is an alternative spawning habitat for P. tenuicorpa in the absence or with a lack of cracks.The relatively low spawning rate of P. tenuicorpa is in contrast to that of P. nigra, belonging to the same genus. P. nigra is only distributed in the upper reaches of the Geum River. Previous studies have reported that the spawning of P. nigra is only dependent on brood parasitism in the nests of C. herzi [P. nigra in the nests of C. herzi was the highest during the first 1–2 days after the nest formation of C. herzi and gradually decreased thereafter. This behavior possibly aimed to align the spawning period (average 10 days) of P. nigra with the nest protection period (14 days for C. herzi’s spawning period) by C. herzi males. When the C. herzi eggs hatch after 14 days, if the C. herzi male is no longer protecting the nest, all the eggs of C. herzi or P. nigra that have not hatched by that time will be consumed by predatory fish  [P. nigra to evolve so as to utilize spawning habitats other than the nests of C. herzi, just like P. tenuicorpa. Therefore, the nest of C. herzi was the most efficient spawning site for P. nigra in the upper reaches of the Geum River.This spawning habitat selection of C. herzi ,44. The cephala; ,45). In P. tenuicorpa in the nests of C. herzi were the highest on the first day after the nest formation of C. herzi, similar to that of P. nigra, and gradually decreased thereafter. However, while the invading and spawning rate of P. nigra in the nests of C. herzi was more than 95%, that of P. tenuicorpa was relatively low, at less than 35%, in four streams , excepting Gapyeong Stream. This implied that the brood parasitism rate of P. tenuicorpa in the nests of C. herzi, distributed in the upper reaches of the Han River, was lower than that in the Geum River. The upper reaches of the Han River were abundant in unspoiled nests not spawned by P. tenuicorpa. As an exception, in the case of Gapyeong Stream, the number of cracks was lower than that in other streams; therefore, the density of P. tenuicorpa using the nests of C. herzi as an alternative spawning habitat increased. The invasion of the nests of C. herzi was attempted by P. tenuicorpa during the first 5 to 6 days after nest formation by C. herzi, and it did not occur after 7 days. This was also presumed to be due to attempts to match the spawning period of P. tenuicorpa with the nest protection period by C. herzi males, as described for P. nigra above.The invading and spawning rates of P. tenuicorpa was due to the preference for and efficiency of abundant cracks smaller than 5 cm as spawning habitats in this area. Although P. tenuicorpa can use both the nests of C. herzi and cracks larger than 6 cm as spawning habitats, both of these spaces are vulnerable to disturbances such as predation, making it difficult to ensure successful hatching and smooth population growth. Since Pseudobagrus sp. and O. platycephala, which utilize the eggs of P. tenuicorpa as a food source, are frequently distributed in the river basin of South Korea, most of the eggs of P. tenuicorpa spawned in cracks larger than 6 cm are consumed and fail to lead to smooth population growth. In the river basins located in South Korea, cracks smaller than 5 cm are the most abundant in the upstream area of the Han River; therefore, P. tenuicorpa has no choice but to distribute based on the upstream area of the Han River. In freshwater ecosystems, the habitat fragmentation of certain species is generally caused by physical barriers , but in the case of P. tenuicorpa, the species cannot disperse to other areas due to the high efficiency of spawning habitats there.From the study results, we concluded that the geographical distribution limit  of P. tenuicorpa distributed around the upper reaches of the Han River evolved from P. herzi. Pungtungia herzi spawns in various spaces, such as giant rocks, gravel, stone cracks, and fish nests [P. herzi requires an efficient spawning habitat to secure continuous population growth. We suspected that P. tenuicorpa is a species derived from P. herzi, with a small size. Small-sized P. herzi accidentally use the narrow cracks of rocks (<5 cm) as a spawning habitat, and the individuals who have succeeded in hatching have continuously reproduced over generations. Pseudopungtungia tenuicorpa is the only fish species that can enter cracks as small as 5 cm in the upper reaches of the Han River. The environmental characteristics of the upper reaches of the Han River, which are abundant in cracks smaller than 5 cm, support P. tenuicorpa to evolve. The nest of C. herzi is also an appropriate spawning habitat, since the males protect the nest from predators, hence keeping the eggs safe. In the upper reaches of the Geum River, P. nigra is a species of fish that actively utilizes the advantages of the nests of C. herzi. Since P. nigra rarely chooses a spawning habitat other than the nests of C. herzi, most of the nests of C. herzi constructed in the upper reaches of the Geum River are spawned by P. nigra. Nevertheless, there are still a few spawning habitats that can replace the nests of C. herzi in the upper reaches of the Geum River (in terms of efficiency and successful hatching). However, if cracks were abundant in the upper reaches of the Geum River, P. nigra would not have adopted a strategy of using the nests of C. herzi as a spawning habitat. Based on these considerations, P. nigra can be presumed to be a species evolved from stone meat, similar to P. tenuicorpa.We assumed that sh nests ,47, but sh nests . Based oP. tenuicorpa in cracks smaller than 5 cm was possible through avoidance of the sharing of spawning habitats with other fish species as much as possible. In the upper reaches of the Han River, P. herzi uses spawning habitats in a manner similar to P. tenuicorpa , but the species do not overlap each other in these spawning habitats. Of course, it is possible that P. herzi avoided cracks and the nests of C. herzi spawned by P. tenuicorpa, but P. herzi spawned in a more diverse area than P. tenuicorpa and was not sensitive enough to avoid the area spawned by P. tenuicorpa [C. herzi is used as a common spawning habitat for P. nigra and C. herzi [P. tenuicorpa avoids spaces that are spawned by P. herzi. Instinctively avoiding spaces spawned by other fish may be a strategy used to explore cracks smaller than 5 cm. Since most of the cracks spawned by P. herzi are more than 6 cm in dimension, and cracks smaller than 5 cm are only spawned by P. tenuicorpa, the strategy of avoiding cracks spawned by P. herzi is effective for finding cracks smaller than 5 cm. Such a behavior is also believed to occur in the case of C. herzi nests. Similar to cracks, we did not find any nest of C. herzi where the eggs of P. tenuicorpa and P. herzi were found at the same time. This was possibly because cracks smaller than 5 cm were the preferred spawning habitat of P. tenuicorpa.The efficient spawning of nuicorpa . PreviouC. herzi . This fiP. tenuicorpa.Such a behavior of spawning alone in a location can be found in the cuckoo catfish of Lake Tanganyika. The cuckoo catfish is parasitic, breeding in the mouths of cichlids, which coexist in Lake Tanganyika ,50. HowePseudopungtungia tenuicorpa was registered as a new fish species in 1980. In South Korea, fish monitoring has been conducted for a long time, and clearly, no P. tenuicorpa was recorded in the upper reaches of the Han River before 1980. The morphological characteristics of P. tenuicorpa are clearly different from those of P. herzi or P. nigra, and therefore, the probability of an error in species identification is also low. Subsequently, the P. tenuicorpa distributed in the upper reaches of the Han River was recognized as a species that recently divided from P. herzi. This resulted from the unusual environmental characteristics of abundant cracks in the upper reaches of the Han River. The presence of cracks smaller than 5 cm contributes greatly to the continuous population growth of P. herzi. This is similar to the case of P. nigra in the upstream area of the Geum River, where the conservation of the nests of C. herzi will be important.P. tenuicorpa in the upper reaches of the Han River has been damaged due to disturbance from anthropogenic factors. The inflow of pollutants and damage to sediment materials due to various recreation activities in this area are damaging the cracks or inducing the clogging of cracks. The decline in or extinction of P. tenuicorpa in this area would mean the extinction of the species in South Korea, since this is a species that is only distributed in the upper reaches of the Han River. Although the landscape characteristics of the area have high aesthetic value for humans, minimizing the anthropogenic disturbance caused in the area for the continuous conservation of P. tenuicorpa is highly recommended. In the future, it will be necessary to consider a plan to artificially create cracks smaller than 5 cm for the spawning and growth of P. tenuicorpa.However, recently, the habitat of P. tenuicorpa. Since cracks smaller than 5 cm are difficult for fish other than P. tenuicorpa to access, the eggs of P. tenuicorpa spawned in such cracks are effectively protected from predators. The high spawning efficiency of P. tenuicorpa in these cracks has greatly contributed to their continued population growth in the upper reaches of the Han River. Pseudopungtungia tenuicorpa has been designated as an endangered species in Korea due to its geographical distribution limit and low population size in the upper reaches of the Han River; suitable management strategies are required for the continuous conservation of the species. Currently, various anthropogenic activities (recreation and dredging) in this area are affecting and damaging the cracks in the upper reaches of the Han River. Since this river is of high value as a habitat for various fish species in addition to P. tenuicorpa, the options of excluding anthropogenic activities from this area as much as possible and restoring the cracks that have already been lost should be considered seriously.We concluded that the abundance of cracks smaller than 5 cm in the upper reaches of the Han River induced unique spawning behavior and geographical distribution restrictions in


Biol. Open (2020) 9, bio051680. doi:10.1242/bio.051680.There were errors published in In The corrected and original figures are shown below. Both the online full-text and PDF versions of the article have been updated. The authors apologise to readers for this error, which does not impact the results or conclusions of this paper.


Helicobacter pylori (H. pylori) infection is ubiquitous worldwide, with prevalence rates of greater than 70% in Africa. Symptomatic patients present with foregut gastrointestinal symptoms which can be readily diagnosed with standardized non-invasive or invasive tests. The biggest challenge, however, is in the management of this condition with rising antimicrobial resistance rates to most of the antibiotics recommended for therapy. This is a problem worldwide, but more specifically in Africa, where the socio-economic and political climate is such that eradication of this organism seems impossible. Furthermore, the recommended antimicrobial susceptibility testing for drug resistance is not widely available in Africa due to the lack of infrastructural as well as human resources. With the widespread unregulated use of antibiotics in some parts of Africa, the figures of antimicrobial resistance are likely to soar. In the face of these significant challenges, this ‘perspectives’ article aims to address the issue of antimicrobial resistance in Africa, by providing achievable and targeted goals to curb the spread of infection and rising antimicrobial resistance. Helicobacter pylori (H. pylori) is the most common infection in the world, with 4.4 billion people infected [H. pylori prevalence in Africa is limited, two recent meta-analyses reported prevalences of at least 70%, representing the highest pooled rates worldwide [H. pylori has declined in developed nations [infected . Althougorldwide ,2. Altho nations ,3, it is nations . This maH. pylori has an affinity for the gastric mucosa, causing universal inflammation [H. pylori is associated with gastritis, duodenitis, peptic ulcer disease [H. pylori is complex and fraught with challenges in developed nations [H. pylori.ammation . Althoug disease ,7 and va disease . Its mos disease , which h disease ,11. Give disease ,13. Whil nations , but arg nations . This haH. pylori is the alarming rates of widespread antimicrobial resistance to most of the recommended antibiotics that form the backbone of H. pylori therapy [H. pylori in Africa under serious threat. Additionally, the lack of AST facilities contributes to the practice of empiric antimicrobial therapy even after failed second line therapy. Therefore, with competing priorities of eradicating H. pylori vis-à-vis observing prudent prescribing practices and choosing effective antibiotics to reduce the rate of antibiotic resistance, quo vadis? How do we in Africa move forward? This article aims to interrogate the recommended therapies for H. pylori infection and antimicrobial resistance including AST in terms of its relevance and applicability in Africa, and to suggest practical and achievable approaches to curb this infection. It is hoped that these recommendations will afford clinicians in Africa some hope of adequately managing H. pylori to prevent complications.Unquestionably, however, the most concerning and challenging issue in the management of  therapy ,17. The  therapy . FurtherH. pylori [H. pylori eradication in Africa and highlights the need for better-designed observational or randomized control trials of available therapies to determine best practices in each region. Second-line therapy is determined by prior choices for first-line therapy, drug availability and antimicrobial resistance. If BQT fails, empiric flouroquinolone (triple or quadruple) therapy or high-dose PPI-amoxicillin therapy is recommended, assuming low flouroquinolone resistance [Several international guidelines from across the globe have been published to guide the diagnosis and management of . pylori ,19,20. N. pylori , is stro. pylori . First-l. pylori . In Afri. pylori . This stsistance . If CTT sistance . Althougsistance . Mountinsistance . Moreovesistance  lending sistance . In termsistance ,26, althsistance . Howeversistance . Rifabutsistance , hence iH. pylori management has been evolving since the discovery of the organism, and indeed continues to evolve. One of the most striking and concerning shifts over the last decade has been increasing antimicrobial resistance [H. pylori [H. pylori antimicrobial resistance is currently so alarming that in 2018, the WHO listed clarithromycin-resistant H. pylori as a high-priority organism for the development of new antibiotics [The landscape of sistance . Clarithsistance . Globallsistance ,  of the importance of completing the course of therapy and be warned of potential side effects. In every case, it cannot be overemphasized that patients must be reviewed for symptoms and confirmation of eradication after at least 30 days post-therapy. In Africa, due to the lack of non-invasive testing in some areas, confirmation of eradication is not always done. This poses significant challenges because of the risk of potentiating antimicrobial resistance where treatment success or failure is not documented.The comprehensive management of H. pylori diagnosis and treatment within geographic locations. This may involve sending specimens outside of the region of the treating hospital to neighbouring regions where these facilities exist. To facilitate this, treating clinicians, through their gastroenterology societies, need to reach out to scientists and microbiologists through publications, research platforms and even social media platforms, to enhance awareness. To this end, a group of gastroenterologists and scientists from eight African countries formed the African Helicobacter and Microbiota Group in 2021 [H. pylori management, including research activities, in Africa. This will facilitate collaborations, so that silos are broken down. It is through these efforts that we can consolidate our efforts and reduce redundancy, such that we can collectively apply for funding that will facilitate the setting up of patient and biorepository registries, as well as identify existing laboratories as centralized facilities for the performance of AST. These laboratories can be capacitated to provide training to newly identified laboratories so that this work can be expanded. The aim is to have at least one AST laboratory in each region of Africa . Through this centralized mechanism, using unified protocols, over time, we will be able to accurately report antimicrobial resistance rates across Africa, both between and within regions. In parallel, another practical and short-term strategy would be for Africa to exploit the infrastructure borne out of the COVID-19 pandemic by performing real-time PCR. In this context, it is also possible that H. pylori reagents can be accessed at a cheaper price for AST since the pandemic. This strategy in the long-term will be fortified by a systematic tracking and reporting of practices , H. pylori eradication rates and related outcomes. This information would be a powerful tool in the fight against antimicrobial resistance in that it will bring to the forefront the possibility of an African H. pylori guideline that is context-specific, but, more importantly, evidence-based. These efforts combined will increase the likelihood of successful eradication of this infection. If second-line treatment fails, AST is recommended. As demonstrated, available data in Africa is sparse, and thus not representative. Even when considering available data, there is wide variability in practice, antibiotic resistance rates and availability of antibiotics. Therefore, the most rational and practical approach is to standardize  in 2021 , akin toH. pylori [While it is not practical to recommend population-wide screening, or screening of all family members living in the same household as patients, it is not unreasonable to suggest screening for those with symptoms within families of patients. This could simply involve a screen for symptoms, and, if present, using a non-invasive tool for diagnosis. In Africa, mainly a stool antigen test, and in a few centres, a UBT, are used for non-invasive testing for . pylori . Family H. pylori have been ignored. Both conditions currently have the least public-health-initiative footprint; and yet have very serious complications. The time has come to put H. pylori on the agenda as a serious infection. We need to engage politicians, policy makers, medical regulatory bodies, drug companies, research funding organizations and community leaders to increase awareness around H. pylori infection. We need educational campaigns, not just for healthcare providers, but for patients, too, so that we can collectively take ownership of infection control, e.g., simple measures such as handwashing, improved hygiene and responsible use of antibiotics. Other risk-reducing practices, such as avoiding non-steroidal anti-inflammatory drugs (NSAIDS), testing and treating for H. pylori in patients on long-term aspirin, anticoagulants, steroids, etc., and reducing alcohol intake and smoking, are important. These need to be included in the comprehensive management of this disorder. At a wider scale, all governments, through bodies such as the African Union , can be made conscious of the need for improved infrastructure in Africa, to improve the general living conditions of people. This will improve control of the increasing antimicrobial resistance, not just for H. pylori, but for many other bacterial infections that are so common in Africa. Unfortunately, at this stage, neither a vaccine nor probiotics are available in clinical practice, and thus these cannot be included in the armamentarium in the preventative fight against H. pylori.There are many stakeholders in the control of this infection. The burden of infectious disease over decades in Africa has been due to HIV, TB and malaria, with justified focus on these. Meanwhile, infections such as hepatitis B and H. pylori infection is not only the most common infection worldwide, but is a group 1 carcinogen. In 20% of patients with symptomatic infection, it is associated with significant morbidity and mortality. Although the diagnosis of H. pylori is standardized and relatively easy to make, treatment and eradication of the organism is fraught with significant challenges, the most concerning being antimicrobial resistance. Although AST facilities are becoming more commonplace in developed nations, this is certainly not the case in Africa, and with the injudicious use of antibiotics, antimicrobial resistance is at crisis point. The immediate priority should be to effectively treat those infected, scale up preventative measures, and involve patients, the medical and scientific professionals, as well as other key stakeholders to combine efforts to successfully eradicate H. pylori. There has been a constant evolution in our understanding of H. pylori; as such, the usual strategies for managing bacterial infections are proving not to be sufficient. In this regard, novel approaches are needed to tackle H. pylori.


Viola epipsila Ledeb., V. palustris L.) are endangered because the ranges of both species are drastically decreased due to global climatic changes. Their disappearance might also be the result of the formation of interspecific hybrids which can replace the parent species. The study of such species is particularly important because they might be considered as indicators of anthropogenic changes occurring in peatlands and their disappearances. The taxonomic situation of studied species is intricate, and the presence of interspecific hybrids and putative introgressants [V. pubifolia (Kuta) G. H. Loos ] makes it even more complicated. The main goal of our study was to reconstruct the origin of V. pubifolia and its genetic relatedness to both putative parental species—V. palustris and V. epipsila—using advanced molecular methods. The taxonomic problem has been finally solved. We found no basis to separate V. pubifolia as a species in its own right because its morphological characters as well as genetic variation fall well within the range of variability of V. palustris. We have also concluded that the low genetic differentiation and heterozygosity of V. epipsila in Europe might be a cause of the reduced tolerance of this species to changing environmental conditions and can possibly lead to its extinction.The wetland violets of Central and Northern Europe  G. H. Loos , interspecific hybrids, and putative introgressants. The genetic affinity of V. pubifolia to V. palustris, and their shared origin via hybridization followed by polyploidization, were confirmed using inter simple sequence repeat (ISSR) markers, restriction site-associated DNA sequencing (RAD-Seq), and a low-copy nuclear gene, GPI, which encodes glucose-6-phosphate isomerase. The other taxa of subsect. Stolonosae were not identified as putative parents of V. pubifolia by GPI. Our analyses indicated that V. pubifolia can be included in the morphological and genetic variation of V. palustris. The ISSR, RAD-Seq, and genome size value separated well V. palustris from V. epipsila and hybrids. The results also reopen the discussion on intraspecific variation in the context of taxa ranks and species concepts. The reduced tolerance of V. epipsila in Europe to changing environmental conditions might result from low genetic differentiation and heterozygosity, as well as the increased number of interspecific hybrids , and eventually can possibly lead to its extinction. The disappearance of populations/individuals of this species may indicate anthropogenic changes occurring in peatlands.In Europe, the  Viola L. is one of the largest angiosperm genera, comprising ca. 658 recognized species; it consists of two sub-genera, 31 sections, and 20 subsections. It is distributed mainly in the temperate zones of both hemispheres and it occupies a high diversity of habitats and elevations .,30.ViolaV. pubifolia and V. palustris were harvested randomly from plants growing in natural sites. Thirty samples of V. pubifolia and one sample of V. palustris were taken from plants located in the experimental garden , three V. epipsila , three hybrids , and three V. pubifolia  samples were used from the previous research , V. occidentalis, V. macloskeyi, V. renifolia, V. principis, V. vaginata, V. selkirkii, V. mirabilis, V. rubella, V. pusilla, and V. congesta were taken from GenBank accessions , followed by genome duplication [V. palustris [V. palustris, is the result of the knockout or silencing of the B homoeolog. Polyploidization induces genetic and epigenetic processes, including DNA sequence elimination and gene silencing, contributing to the functional diversification or subfunctionalization of duplicated genes, and the genetic and cytological diploidization of allopolyploids. Epigenetic modifications may produce adaptive epimutations and novel phenotypes [This great morphological variability of lication . In sexualustris , new genenotypes ,75,76.V. pubifolia individuals observed at CH or CL blooming can lead to erroneous classification to V. epipsila or hybrid V. epipsila × V. palustris [The hairy lower leaf surface and the location of bracts in the middle or even above the middle of the pedicel of alustris ,10.V. palustris but not a hybrid, which is sterile. It should also be taken into account that the shape and hairiness of the leaf blade are seasonally variable; there are also differences between leaves developing on stolons and on rhizomes (stolon leaves are more acute and with a more open sinus). Herbarium specimens in the vegetative stage (only leaves and rhizomes) are impossible to be correctly identified in the V. epipsila—V. palustris group .It is recommended to describe and classify specimens in the field during the period of CH or CL fruiting. Normally developed, the seed-filled capsules indicate Viola as important for biodiversity and nature conservation. The increased number of interspecific hybrids , with a simultaneous reduction in the number of V. epipsila and/or V. palustris individuals or populations V. pubifolia fall well within the range of variability of V. palustris.Morphological characters of (2)V. pubifolia is V. palustris, based on ISSR, GPI, and RAD-Seq.Genetically, (3)V. epipsila confirmed by RAD-Seq might explain its low tolerance to changing environments and the risk of extinction.The low genetic diversity and heterozygosity of selected Central and Northern European populations of (4)V. epipsila populations and the disappearance of its natural habitats suggest that in some areas it might require effective conservation strategies.The declining number of


Journal of Clinical Hypertension 24: 1390−1391. https://doi.org/10.1111/jch.14353.Ortiz, M.S. (2022), Inconsistencies in the sensitivity and specificity values in a review paper published in Journal of Clinical Hypertension.This note is for the above Letter to the Editor, published online on August 21, 2021 on Wiley Online Library (wileyonlinelibrary.com), and has been published by agreement between the Journal's Editor‐in‐Chief, Dr Ji‐Guang Wang, and Wiley Periodicals, LLC. In the above Letter to the Editor, concerns were raised regarding inconsistencies in the sensitivity and specificity values in an article by Morisky et al. published in the