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Pyrenophora tritici-repentis causes tan spot, an important foliar disease of wheat. A collection of P. tritici-repentis isolates from Tunisia, located in one of the main secondary centers of diversification of durum wheat, was tested for phenotypic race classification based on virulence on a host differential set and for the presence of the necrotrophic effector (NE) genes ToxA, ToxB, and toxb by PCR analysis. While races 2, 4, 5, 6, 7, and 8 were identified according to their virulence phenotypes, PCR testing indicated the presence of “atypical” isolates that induced necrosis on the wheat differential ‘Glenlea,’ but lacked the expected ToxA gene, suggesting the involvement of other NEs in the P. tritici-repentis/wheat interaction. Genetic diversity and the P. tritici-repentis population structure were explored further by examining 59 Tunisian isolates and 35 isolates from Algeria, Azerbaijan, Canada, Iran, and Syria using 24 simple sequence repeat markers. Average genetic diversity, overall gene flow, and percentage polymorphic loci were estimated as 0.58, 2.09, and 87%, respectively. Analysis of molecular variance showed that 81% of the genetic variance occurred within populations and 19% occurred between populations. Cluster analysis by the unweighted pair group method indicated that ToxB– isolates grouped together and were distantly related to ToxB+ isolates. Based on Nei’s analysis, the global collection clustered into two distinct groups according to their region of origin. The results suggest that geographic origin and the host specificity imposed by different NEs can lead to differentiation among P. tritici-repentis populations.

Background. The prevalence of giardiasis among children is 355 cases per 100,000, with 80 % of cases of infestation occurring in children under the age of 14 years. The urgency of the high prevalence of protozoal pathology in children is determined by the fact that its manifestations are often masked under various pathological conditions. The purpose was to study the incidence, predictors and criteria for the diagnosis of giardiasis in children. Materials and methods. Research design is a controlled randomized trial. The study included children suffering from giardiasis (n = 55), aged 3 to 17 years. The comparison group consisted of 30 apparently healthy children. A survey of patients and collection of material performed at the outpatient and gastroenterology departments of the MHI “Chernivtsi Regional Clinical Hospital” during 2014–2016. Laboratory and instrumental investigations were conducted in Ukrainian-German laboratory “Bukintermed” and children’s gastroenterology center “Digest”. Indirect (serological one, macroscopy of the duodenal mucosa in endoscopic examination, indirect signs of giardiasis in duodenal biopsy samples) and direct methods were used (microscopy of the feces, duodenal contents, detection of vegetative forms of lamblia in duodenal biopsy samples during histological examination). A parent survey was conducted. Results. A positive result was obtained in 29 out of 565 subjects surveyed for lamblia (5.1 %). More often, additional examination was required for the presence of giardiasis in children suffering from chronic diseases of the upper gastrointestinal and hepatobiliary system, allergic reactions, intestinal dysbiosis and persistent eosinophilia. Children affected with giardiasis were more likely to have a concomitant illness than the children of the comparison group (p < 0.05). In 41.4 % of cases, children lived in conditions that were not satisfactory, did not always follow the rules of sanitary hygiene. We have established antenatal (health during pregnancy, the course of pregnancy, the presence of occupational hazards), intranatal (premature birth, stimulation of labor and cesarean section, birth trauma and asphyxia in children) and postnatal adverse factors (duration of breastfeeding, concomitant pathology). All histological features can be used as possible criteria for giardiasis in children, but with the highest probability, the following sign can be used — the presence of villous epithelium desquamation areas with simultaneous dominance of eosinophils and mast cells in the infiltrate. Conclusions. The incidence of giardiasis in children and adolescents is 5.1 %. Anamnestic factors (social, hygienic, medical and biological ones) established by us can be considered as predictors for the development of giardiasis in children. In the clinical picture of giardiasis, the defeat of the digestive tract dominates — in 96.3 % of children. To improve the accuracy of giardiasis diagnosis in children, it is necessary to use a set of direct and indirect methods of the diagnosis.

Resistance to late blight caused by Phytophthora infestans clonal lineage US-23 in 217 old and modern potato cultivars was evaluated in field trials in 2016 and 2017 in Pennsylvania. Significant differences in resistance were found among these cultivars (P < 0.0001). Significant interaction between cultivars and environments was found (P < 0.0001). The values of relative area under the disease progress curve ranged from 0 to 0.5841 in 2016 and from 0 to 0.5469 in 2017. Broad-sense heritability of late blight resistance was estimated to be 0.91, with a 95% confidence interval of 0.88 to 0.93. Cluster analysis classified the cultivars into five groups: resistant, moderately resistant, intermediate, moderately susceptible, and susceptible. Thirty cultivars showing resistance and 32 cultivars showing moderate resistance were identified. The 217 cultivars were also evaluated for foliar maturity, tuber yield, and resistance to early blight caused by Alternaria solani. A few cultivars with late blight resistance independent of late maturity were found. Late blight resistance and early blight resistance were positively correlated, and 17 cultivars possessed resistance to both diseases. Yield trade-off associated with late blight resistance was not observed among the cultivars in the absence of disease pressure.

The soybean cyst nematode (SCN) Heterodera glycines continues to be a major threat to soybean production worldwide. Morphological discrimination between SCN and other nematodes of the Heterodera schachtii sensu stricto group is not only difficult and time-consuming but also requires high expertise in nematode taxonomy. Molecular assays were developed to differentiate SCN from sugar beet cyst nematode (SBCN) and other nematodes and to quantify SCN directly from DNA extracts of field soils. SCN- and SBCN-specific quantitative real-time PCR (qPCR) primers were designed from a nematode-secreted CLAVATA gene and used for these assays. The primers were evaluated on the basis of target specificity to SCN or SBCN using DNA from 20 isolates of SCN and 32 isolates of other plant-parasitic nematodes. A standard curve relating threshold cycle and log values of nematode numbers was generated from artificially infested soils and was used to quantify SCN in naturally infested field soils. There was a high correlation between the SCN numbers estimated from naturally infested field soils by conventional methods, and the numbers quantified using the SYBR Green I-based qPCR assay. The qPCR assay is highly specific and sensitive and provides improved SCN detection sensitivity down to 1 SCN egg in 20 g of soil (10 eggs/200 g soil). This assay is useful for efficient detection and quantification of SCN directly from field soil. Species-specific conventional PCR assays were also developed each for SCN and SBCN, alongside a qPCR assay that simultaneously discriminates SCN from SBCN. These assays require no expertise in nematode taxonomy and morphology, and they may serve as useful diagnostic tools in research, diagnostic laboratories, and extension services for SCN management. Sensitive and accurate detection and quantification of SCN are essential for recommending effective management measures against SCN. We also investigated the impact of soil texture and nematode life stage on molecular quantification of SCN.

Fire blight, caused by the bacterium Erwinia amylovora, is one of the most important diseases of apple. The antibiotic streptomycin is routinely used in the commercial apple industries of New York (NY) and New England to manage the disease. In 2002 and again, from 2011 to 2014, outbreaks of streptomycin resistance (SmR) were reported and investigated in NY. Motivated by new grower reports of control failures, we conducted a follow-up investigation of the distribution of SmR and E. amylovora strains for major apple production regions of NY over the last 6 years (2015 to 2020). Characterization of clustered regularly interspaced short palindromic repeat (CRISPR) profiles revealed that a few “cosmopolitan” strains were widely prevalent across regions, whereas many other “resident” strains were confined to one location. In addition, we uncovered novel CRISPR profile diversity in all investigated regions. SmR E. amylovora was detected only in a small area spanning two counties from 2017 to 2020 and was always associated with one CRISPR profile (41:23:38), which matched the profile of SmR E. amylovora, discovered in 2002. This suggests the original SmR E. amylovora was never fully eradicated and went undetected because of several seasons of low disease pressure in this region. Investigation of several representative isolates under controlled greenhouse conditions indicated significant differences in aggressiveness on ‘Gala’ apples. Potential implications of strain differences include the propensity of strains to become distributed across wide geographic regions and associated resistance management practices. Results from this work will directly influence sustainable fire blight management recommendations for commercial apple industries in NY state and other regions.

Fusarium wilt (FW), caused by Fusarium oxysporum f. sp. vasinfectum (Atk.) W.C. Snyder & H.N. Hans (FOV), is one of the most destructive diseases of cotton (Gossypium spp.) worldwide. FOV race 4 (FOV4) is a highly virulent nominal race of this pathogen and a significant threat to cotton production in the western and southwestern USA and, potentially, the entire Cotton Belt. A field survey to identify FOV4 was performed in three southern counties of New Mexico in 619 cotton fields from 2018 to 2020. From 132 samples of cotton plants that exhibited wilt symptoms, Fusarium spp. were the most frequently isolated group of fungal species, with an isolation frequency of 57.4%. Eighty-four Fusarium spp. isolates were subsequently characterized by a DNA sequence analysis of three genes, EF-1α, PHO, and BT, encoding for translation elongation factor, phosphate permease, and β-tubulin, respectively. Forty-two isolates from 10 cotton fields were identified as FOV4 and confirmed with a positive 500-bp fragment diagnostic for FOV4. Twenty-six (62%) of the 42 FOV4 isolates were T type and the remainder (38%) were null type with and without a Tfo1 insertion in PHO, respectively. Each FOV4-infested field contained the same FOV4 genotype. Ten representative FOV4 isolates (one each from the 10 FOV4-infested fields) were evaluated for their pathogenicity on resistant Pima PHY 841 RF and susceptible Upland PHY 725 RF at 7, 14, 21, and 28 days after inoculation under temperature-controlled conditions at 21 to 22°C. Based on the disease severity rating, mortality rate, and area under the disease progress curve value, all 10 isolates were pathogenic to both cotton cultivars and differed in virulence; four isolates of the T genotype as a whole were more virulent than the six isolates of the N genotype. PHY 841 RF had significantly higher levels of resistance than PHY 725 RF to all FOV4 isolates. The results provide the first comprehensive account of the occurrence, distribution, and virulence of FOV4 in cotton production in New Mexico and will be useful for developing an effective strategy to manage FW in the state of New Mexico and the entire western and southwestern Cotton Belt.

Within the framework of a taxonomic revision of Hyacinthaceae subfamily Ornithogaloideae, we present two new combinations in Eliokarmos for South African species, and five new combinations in Loncomelos for taxa occurring in Iran and Iraq. Details on the concerned types are given, and a new lectotype is designated. This work was partly supported by H2020 Research and Innovation Staff Exchange Programme of the European Commission, Project 645636: 'lnsect-plant relationships: insights into the biodiversity and new applications' (FlyHigh), and the grants ACIE18-03 and UAUSTI18-02 from University of Alicante.

South Tyrol (northern Italy) harbors one of the largest interconnected apple farming areas in Europe, contributing approximately 10% to the apple production of the European Union. Despite the availability of sophisticated storage facilities, postharvest diseases occur, one of which is bitter rot of apple. In Europe, this postharvest disease is mainly caused by the Colletotrichum acutatum species complex. This study aimed to characterize the Colletotrichum spp. isolated from decayed apple fruit collected in 2018 and 2019 in South Tyrol. The characterization of Colletotrichum spp. was accomplished based on multilocus DNA sequences of four different genomic regions—actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), histone H3 (HIS3), and the internal transcribed spacer (ITS) region—as well as morphological and pathogenicity assessment. A phylogenetic analysis based on multilocus DNA sequences showed that the isolates obtained from apples with symptoms of bitter rot belonged to the species Colletotrichum godetiae and Colletotrichum fioriniae, which are part of the Colletotrichum acutatum species complex. A third species isolated from apples belonging to the same species complex, Colletotrichum salicis, was described in this area. Moreover, the Colletotrichum isolates found in this study proved to be virulent on Cripps Pink, Golden Delicious, and Roho 3615/Evelina. To the best of our knowledge, C. godetiae and C. fioriniae have so far never been mentioned as postharvest pathogens of apple in Italy, although the reanalysis of samples collected in the past indicates that these pathogens have been occurring in Italy for at least a decade. So far, bitter rot seems to play a minor role as a postharvest disease in South Tyrol, but it was disproportionately represented on a few scab-resistant apple cultivars, which are increasingly planted in organically managed orchards. Considering that the expansion of organic apple production and the conversion to new potentially Colletotrichum-susceptible cultivars will continue, the present study represents an important contribution toward a better understanding of bitter rot in this geographic area. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY 4.0 International license .

Background: Seasonal outbreaks of acute encephalitis syndrome (AES) have been reported especially in the pediatric population with a high case fatality rate in Eastern Uttar Pradesh, India. Orienti...

The ability to detect and quantify aerially dispersed plant pathogens is essential for developing effective disease control measures and epidemiological models that optimize the timing for control. There is an acute need for managing the downy mildew pathogens infecting cucurbits and hop incited by members of the genus Pseudoperonospora (Pseudoperonospora cubensis clade 1 and 2 isolates and Pseudoperonospora humuli, respectively). A highly specific multiplex TaqMan quantitative polymerase chain reaction (PCR) assay targeting unique sequences in the pathogens’ mitochondrial genomes was developed that enables detection of all three taxa in a single multiplexed amplification. An internal control included in the reaction evaluated whether results were influenced by PCR inhibitors that can make it through the DNA extraction process. Reliable quantification of inoculum as low as three sporangia in a sample was observed. The multiplexed assay was tested with DNA extracted from purified sporangia, infected plant tissue, and environmental samples collected on impaction spore traps samplers. The ability to accurately detect and simultaneously quantify all three pathogens in a single multiplexed amplification should improve management options for controlling the diseases they cause.

Matricaria chamomilla flower extract was used as a biocompatible material for synthesis of zinc oxide nanoparticles (ZnONPs). The synthesized NPs were evaluated for their antibacterial potential in vitro and in vivo against the Gram-negative bacterium Ralstonia solanacearum, which causes devastating bacterial wilt disease in tomato and other crops. Synthesized ZnONPs were further analyzed by UV-visible spectroscopy, Fourier transform infrared spectroscopy, x-ray diffraction, transmission electron microscopy, and scanning electron microscopy with energy-dispersive spectroscopy. The synthesized polydisperse ZnONPs were found to be in the size range of 8.9 to 32.6 nm, and at 18.0 µg ml−1 exhibited maximum in vitro growth inhibition of the pathogen R. solanacearum. Scanning electron microscopy analysis of affected bacterial cells showed morphological deformation such as disruption of the cell membrane and wall, and the leakage of cell contents. Results of in vivo studies also showed that application of ZnONPs to the artificially inoculated tomato plants with the pathogen R. solanacearum significantly enhanced the plant growth by reducing bacterial soil population and disease severity as compared with the untreated control. Biosynthesized ZnONPs could be an effective approach to control the bacterium R. solanacearum.

Blackleg of oilseed rape is a damaging invasive disease caused by the species complex Leptosphaeria maculans (Lm)/L. biglobosa (Lb), which is composed of at least two and seven phylogenetic subclades, respectively. Generally, Lm is more virulent than Lb, but under certain conditions, Lb can cause a significant yield loss in oilseed rape. Lb ‘brassicae’ (Lbb) has been found to be the causal agent for blackleg of oilseed rape in China, whereas Lm and Lb ‘canadensis’ (Lbc) were frequently detected in imported seeds of oilseed rape, posing a risk of spread into China. To monitor the blackleg-pathogen populations, a diagnostic tool based on loop-mediated isothermal amplification (LAMP) was developed using a 615-bp–long DNA sequence from Lbb that was derived from a randomly amplified polymorphic DNA assay. The LAMP was optimized for temperature and time, and tested for specificity and sensitivity using the DNA extracted from Lbb, Lbc, Lm, and 10 other fungi. The results showed that the optimal temperature and time were 65°C and 40 min, respectively. The LAMP primer set was specific to Lbb and highly sensitive as it detected the Lbb DNA as low as 132 fg per reaction. The LAMP assay was validated using the DNA extracted from mycelia and conidia of a well-characterized Lbb isolate, and its utility was evaluated using the DNA extracted from leaves, stems, pods, and seeds of oilseed rape. The LAMP assay developed herein will help for monitoring populations of the blackleg pathogens in China and in developing strategies for management of the blackleg disease.

Rice is a key food security crop in Africa. The importance of rice has led to increasing country-specific, regional, and multinational efforts to develop germplasm and policy initiatives to boost production for a more food-secure continent. Currently, this critically important cereal crop is predominantly cultivated by small-scale farmers under suboptimal conditions in most parts of sub-Saharan Africa (SSA). Rice blast disease, caused by the fungus Magnaporthe oryzae, represents one of the major biotic constraints to rice production under small-scale farming systems of Africa, and developing durable disease resistance is therefore of critical importance. In this review, we provide an overview of the major advances by a multinational collaborative research effort to enhance sustainable rice production across SSA and how it is affected by advances in regional policy. As part of the multinational effort, we highlight the importance of joint international partnerships in tackling multiple crop production constraints through integrated research and outreach programs. More specifically, we highlight recent progress in establishing international networks for rice blast disease surveillance, farmer engagement, monitoring pathogen virulence spectra, and the establishment of regionally based blast resistance breeding programs. To develop blast-resistant, high yielding rice varieties for Africa, we have established a breeding pipeline that utilizes real-time data of pathogen diversity and virulence spectra, to identify major and minor blast resistance genes for introgression into locally adapted rice cultivars. In addition, the project has developed a package to support sustainable rice production through regular stakeholder engagement, training of agricultural extension officers, and establishment of plant clinics.

Recently, there has been a significant growth in parasitic invasions among young children. At a younger age, parasitic infestations most often occur “under the mask” of gastroenterological pathology. At this age range, according to the domestic and world literature, functional diseases prevail. In May 2016, at the week of American gastroenterology, the Rome criteria IV were adopted. According to the the Rome criteria IV, as well as in the previous editions, it is suggested to prescribe therapy on the basis of the predominant symptom principle, giving preference to so-called complementary and alternative medicine, and, in particular, phytotherapy as additional therapy for the disease, but all also from the standpoint of evidence. The first such combined phytocomplex, which attracted interest from clinical researchers and patients, is Vormil Phyto — a mix of extracts of thirteen plants. The purpose of our research was the improvement of therapeutic approaches to the correction of the combined functional pathology of the biliary tract and parasitic invasion, as well as unassociated pathology of the biliary tract using phytocomplex Vormil Phyto in the main treatment regimens. Sixty children were divided into three groups: the first group with combined functional disorder of the biliary tract and parasitic infestation; the second group with a functional biliary tract disorder, received the phytocomplex Vormil Phyto; the third group with a functional biliary tract disorder, received traditional therapy. In the course of the research, data were obtained that indicate the effectiveness of the phytocomplex Vormil Phyto, which has not only an antihelmintic effect, but also normalizes the kinetics of the biliary tract that improves appetite rapidly and does not induce dyspeptic symptoms at earlier terms.

Background. Pediatric co-infections are an emerging clinical problem due to their increasing prevalence and tendency to transform a typical clinical presentation of particular diseases. Thereafter, it tangles the accurate estimation of etiology, complicates the management and negatively impacts the outcome. Given the climatic changes, significant migratory flows and international tourism, tropical helminthiases, previously not common in Ukraine, are a real threat to the public health, especially in combination with other pathogens. Materials and methods. We observed a case of strongyloidiasis and salmonellosis in a Ukrainian 5-month-old female infant who had no history of visiting any of the sub- or tropical territory of the globe. The girl came from a socially vulne­rable family and was abandoned by her homeless parents immediately after admission. The girl presented with severe toxic manifestations, diarrhea, developmental delay, moderate-to-severe malnutrition and dehydration, and maculopapular rash on the trunk and lower extremities. Direct light microscopy of feces revealed Str. stercoralis in the number of more than 10 mobile larvae per high-power field, at different stages of development. Bronchial lavage fluid contained no larvae of Str. stercoralis. Fecal culture revealed group D S. enteritidis. Chemotherapy with ceftriaxone IV and oral albendazole resulted in elimination of both pathogens. Conclusions. The given case of S. enteritidis and Str. stercoralis co-infection should be considered as a probable case of autochthonous Str. stercoralis infection, as it was not confirmed by more reliable diagnostic methods (e.g. PCR for Str. stercoralis DNA), and demonstrates a doubtful epidemiological history. Consequently, to improve the diagnosis of endemic parasitic infections, it is necessary to introduce such a verification as compulsory, and mandatory registration of relevant cases in the Ukrainian National System of Epidemiological Surveillance and Biosecurity is required., Актуальность. Сочетанные инфекции в педиатрии являются клинической проблемой, приобретающей актуальность из-за их распространенности и тенденции к изменению типичной клинической картины отдельных заболеваний. Соответственно, это затрудняет точную оценку этиологии, процесс лечения и негативно влияет на результат. Учитывая изменения климата, значительные миграционные потоки и международный туризм, тропические гельминтозы, которые ранее не были распространены в Украине, являются реальной угрозой для здоровья населения, особенно в сочетании с другими патогенами. Материалы и методы. Мы наблюдали случай стронгилоидоза и сальмонеллеза у 5-месячного ребенка, который не посещал никаких суб- или тропических территорий земного шара. Ребенок происходит из социально незащищенных слоев общества и оставлен бездомными родителями сразу после госпитализации. У девочки отмечались выраженные тяжелые токсические проявления, диарея, задержка развития, нарушение питания и обезвоживание от умеренной до тяжелой степени, пятнисто-папулезные высыпания на туловище и нижних конечностях. Прямая световая микроскопия фекалий обнаружила Strongyloides stercoralis в количестве более 10 подвижных личинок в поле зрения, на разных стадиях развития. Промывные воды бронхиального лаважа не содержали личинок Str. stercoralis. Фекальная культура обнаружила Salmonella enteritidis группы D. Химиотерапия цефтриаксоном внутривенно и альбендазолом перорально привела к элиминации обоих патогенов. Выводы. Данный случай смешанной инфекции S. enteritidis и Str. stercoralis может рассматриваться только как вероятный случай аутохтонной инфекции Str. stercoralis, поскольку она не была подтверждена более надежными методами диагностики (например, ПЦР-определение ДНК Str. stercoralis) и демонстрирует сомнительную эпидемиологическую историю. Соответственно, для улучшения диагностики эндемичных паразитарных инфекций необходимо ввести такую верификацию как обязательную и обязательно регистрировать соответствующие случаи в национальной системе эпидемиологического надзора и биологической безопасности., Актуальність. Поєднані інфекції в педіатрії є клінічною проблемою, що набуває актуальності через їх поширеність і тенденцію до зміни типової клінічної картини окремих захворювань. Відповідно, це ускладнює точну оцінку етіології, процес лікування та негативно впливає на результат. Враховуючи зміни клімату, значні міграційні потоки та міжнародний туризм, тропічні гельмінози, які раніше не були поширені в Україні, є реальною загрозою для здоров’я населення, особливо в поєднанні з іншими патогенами. Матеріали та методи. Ми спостерігали випадок стронгілоїдозу та сальмонельозу в 5-місячної дитини, яка не відвідувала будь-якої суб- або тропічної території земної кулі. Дівчина походить із соціально незахищених верств суспільства і була залишена безпритульними батьками відразу після госпіталізації. У дівчини відмічались виражені тяжкі токсичні прояви, діарея, затримка розвитку, порушення харчування та зневоднення від помірного до тяжкого ступеня, плямисто-папульозні висипання на тулубі й нижніх кінцівках. Пряма світлова мікроскопія фекалій виявила Strongyloides stercoralis у кількості понад 10 рухливих личинок у полі зору, на різних стадіях розвитку. Промивна рідина бронхіального лаважу не містила личинок Str. stercoralis. Фекальна культура виявила Salmonella enteritidis групи D. Хіміотерапія цефтріаксоном внутрішньовенно й альбендазолом перорально призвела до елімінації обох патогенів. Висновки. Цей випадок змішаної інфекції S. enteritidis і Str. stercoralis може розглядатися лише як імовірний випадок автохтонної інфекції Str. stercoralis, оскільки вона не була підтверджена більш надійними методами діагностики (наприклад, ПЛР-визначення ДНК Str. stercoralis) і демонструє сумнівну епідеміологічну історію. Отже, для поліпшення діагностики ендемічних паразитарних інфекцій необхідно запровадити таку верифікацію як обов’язкову та обов’язково реєструвати відповідні випадки в національній системі епідеміологічного нагляду та біологічної безпеки.

Alternaria rot caused by Alternaria species is one of the major postharvest diseases of mandarin fruit in California. The aims of this study were to identify these Alternaria species via phylogenetic analyses and morphological characteristics and test their pathogenicity on mandarin. Decayed mandarin fruits exhibiting Alternaria rot symptoms were collected from three citrus fruit packinghouses in the Central Valley of California. In total, 177 Alternaria isolates were obtained from decayed fruit and preliminarily separated into three groups representing three species (A. alternata, A. tenuissima, and A. arborescens) based on the colony characterization and sporulation patterns. To further identify these isolates, phylogenetic analysis was conducted based on DNA sequences of the second largest subunit of RNA polymerase II (RPB2), plasma membrane ATPase (ATPase), and Calmodulin gene regions in combination with morphological characters. Of the 177 isolates, 124 (70.1%) were identified as A. alternata, and 53 (29.9%) were A. arborescens. The isolates initially identified as A. tenuissima based on the morphological characteristics could not be separated from those of A. alternata in phylogenetic analysis and thus considered A. alternata. Pathogenicity tests showed that both Alternaria species were pathogenic on mandarin fruit at both 5°C and 20°C. Our results indicated that two Alternaria species, A. alternata and A. arborescens, were responsible for Alternaria rot of mandarin fruit in California, with A. arborescens causing fruit rot on mandarin being reported for the first time.

Apple powdery mildew, caused by Podosphaera leucotricha, continues to be a challenge in commercial apple orchards in the U.S. Pacific Northwest and worldwide. In this study, P. leucotricha isolates were collected in 2018 and 2019 from two organic (baseline) and eight conventional (exposed) apple orchards in Washington, New York, and Virginia, and assessed for their sensitivity to trifloxystrobin (TRI, n = 232), triflumizole (TFZ, n = 217), and boscalid (BOS, n = 240) using a detached leaf assay. Effective concentrations inhibiting 50% growth (EC50) were not significantly different between baseline and exposed isolates, and ranged from 0.001 to 0.105, 0.09 to 6.31, and 0.05 to 2.18 µg/ml, for TRI, TFZ, and BOS, respectively. Reduction in sensitivity by factors of 105, 63, and 22 to TRI, TFZ, and BOS, respectively, were observed in some isolates, but all isolates were controlled by the commercial label rates of the three fungicides on detached leaves. Sequencing of the cytochrome b (cytb), cytochrome P450 sterol 14α-demethylase (CYP51), and the iron-sulfur protein subunit (SdhB) genes in isolates with high EC50 revealed no mutation previously reported to confer resistance to these fungicides in other fungi, and presence of a group I intron after codon 143 in the cytb gene. Significant (P < 0.001) moderate positive correlations (r = 0.38) observed between sensitivity to TRI and TFZ warrant continuous rotations of fungicides with different modes of action in conventional orchards. The established baseline sensitivities and the molecular markers will help in selecting discriminatory doses and bypassing the challenging in vivo testing for future sensitivity monitoring in P. leucotricha.

Target spot, caused by Corynespora cassiicola, has reemerged as a disease of economic importance in cotton (Gossypium hirsutum L.) in the southeastern United States. Although this pathogen affects other economically important crops, relatively little is known about C. cassiicola on cotton, especially with regard to conditions conducive for disease development and sources of genetic resistance. Therefore, to more efficiently screen cotton genotypes for resistance, a greenhouse protocol is needed. Optimum temperature and conducive leaf wetness duration were determined in growth chamber trials. Temperature range for disease onset and greatest lesion counts occurred between 20 and 28°C. Generally, with ≥24 h of leaf wetness at these temperatures, disease onset was noted as rapidly as 1 day after inoculation on a cultivar that was previously determined to be susceptible to target spot. A mist irrigation system was used to maintain prolonged periods of leaf wetness in the greenhouse. In greenhouse trials, inoculation of cotyledons with 4 × 104 conidia/ml allowed differentiation of five selected genotypes with disease reactions that reflected their field rankings. The current protocol will be useful for evaluating cotton breeding lines for resistance to target spot.

Culex pipiens sensu lato has been documented as West Nile virus (WNV) vector in southeastern Romania. Bucharest, the densely populated capital city of Romania, and the surrounding Ilfov county are WNV hotspots. In this area, the morphologically indistinguishable biotypes of Cx. pipiens, namely pipiens and molestus, are usually differentiated by their behavioral and physiological traits. Their involvement in WNV transmission, as suggested by entomological investigations, was not previously documented for each biotype. We used a Real-Time PCR assay based on CQ11 microsatellite to identify the Cx. pipiens biotypes and their hybrids collected in various habitats in the Bucharest metropolitan area. A sympatric distribution of both biotypes was observed, with a preference of green areas for pipiens, and human settings and animal farmlands for molestus. In the latter habitats, pipiens and molestus were found in mixed aboveground populations. A low number of hybrids was found suggesting existence of reproductive isolation. In subway tunnels molestus was dominant with a higher number of hybrids recorded than aboveground. Blood-engorged mosquitoes were identified to biotype and the blood meal source identified by DNA barcoding. Overall, Cx. pipiens s.l. fed mainly on birds, commonly on house sparrows, collared doves, and blackbirds, which are potential WNV-amplifying hosts. The preference for avian hosts was expressed strongest by pipiens biotype, while molestus was substantially less specific, feeding on avian and mammal hosts with similar frequency, with humans representing 20% of the hosts. Hybrids had a host choice closer to that of molestus. These findings highlight the role of pipiens biotype as enzootic/epizootic vector, and specifically show molestus as the bridge vector for WNV. The pipiens and molestus biotypes show important differences in habitat preferences, including oviposition; these findings demonstrate that targeted mosquito control to limit WNV transmission may be possible.

Plague is caused by a bacterial pathogen (Yersinia pestis) that can infect a wide range of mammal species, but its presence in wildlife is often underappreciated. Using a large-scale data set (n = 44,857) that details the extent of Y. pestis exposure in wildlife, we document exposure in 18 wildlife species, including coyotes (Canis latrans), bobcats (Lynx rufus), and black bears (Ursus americanus). Evidence of plague activity is widespread, with seropositive animals detected in every western state in the contiguous United States. Pathogen monitoring systems in wildlife that are both large scale and long-term are rare, yet they open the door for analyses on potential shifts in distribution that have occurred over time because of climate or land use changes. The data generated by these long-term monitoring programs, combined with recent advances in our understanding of pathogen ecology, offer a clearer picture of zoonotic pathogens and the risks they pose.

Giardia lamblia (syn Giardia duodenalis) is an important protozoan parasite that can cause enterocyte damage and loss of brush border of the epithelial cells in the intestine, resulting in shortening of microvilli and altered epithelial barrier function. Many animals have been detected as the hosts of the G. lamblia. However, the information on the epidemiology and molecular detection of G. lamblia in dairy calves and sika deer in northeastern China is limited. To investigate the prevalence and genotypes of dairy calves and sika deer in northeastern China, a total of 321 fecal samples from dairy calves in Heilongjiang Province and 818 fecal samples from sika deer in four provinces (Jilin Province, Heilongjiang Province, Liaoning Province, and Inner Mongolia Autonomous Region) in China were conducted by PCR methods, between September 2017 and April 2018. The overall prevalence of G. lamblia in dairy calves in Heilongjiang Province and sika deer in the four provinces was 4.98% (16/321) and 0.61% (5/818), respectively. In this study, the point prevalence of Giardia spp. in different factor groups was dissimilar. A total of 16 Giardia spp. positive samples in dairy calves were identified as assemblage E based on the triosephosphate isomerase (tpi), β-giardine (bg), and glutamate dehydrogenase (gdh) genes. Furthermore, two positive samples of assemblage A and three positive samples of assemblage E were identified with gdh and bg genes in the sika deer. Assemblage A was zoonotic genotype of G. lamblia, and assemblage E was identified as the predominant assemblage in dairy calves and sika deer. This study reported the prevalence and genotypes of G. lamblia in dairy calves in Heilongjiang Province and sika deer in four provinces in China. These results provided basic information to understand the epidemiology of G. lamblia in dairy calves and sika deer in China.

Sclerotinia sclerotiorum is a devastating plant pathogen with a broad host range and worldwide distribution. The application of chemical fungicides is a primary strategy for controlling this pathogen. However, under the high selective pressure of chemical fungicides, fungicide resistance has emerged and gradually increased, resulting in the failure to control S. sclerotiorum in the field. Quinofumelin is a novel quinoline fungicide, but its antifungal activities against plant pathogens have been rarely reported. Here, we determined the antifungal activity of quinofumelin against S. sclerotiorum in vitro and in planta. The median effect concentration (EC50) values ranged from 0.0004 to 0.0059 µg ml−1 with a mean EC50 of 0.0017 ± 0.0009 µg ml−1 and were normally distributed (P = 0.402). In addition, no cross resistance was observed between quinofumelin and other fungicides, dimethachlone, boscalid, or carbendazim, which are commonly used to manage S. sclerotiorum. Quinofumelin did not affect glycerol and oxalic acid production of either carbendazim-sensitive or -resistant isolates. Moreover, quinofumelin exhibited excellent protective, curative, and translaminar activity against S. sclerotiorum on oilseed rape leaves. Protective activity was higher than curative activity. Interestingly, quinofumelin inhibited the formation of the infection cushion in S. sclerotiorum, which may contribute to the control efficacy of quinofumelin against S. sclerotiorum in the field. Our findings indicate that quinofumelin has excellent control efficacy against S. sclerotiorum in vitro and in planta as compared with extensively used fungicides and could be used to manage carbendazim- and dimethachlone-resistance in S. sclerotiorum in the field.

Anthracnose is an important disease of grapevines caused by the fungus Elsinoë ampelina. In recent years, there have been regular outbreaks in humid grape-growing regions around the world. Young leaves and berries are reported to be highly susceptible to E. ampelina, but detailed and seasonal development of age-related susceptibility remains unclear. Experiments were conducted under greenhouse and vineyard conditions by inoculating 1- to 19-day-old leaves, flowers, and berries at different phenological stages of three grapevine cultivars (Vandal-Cliche, Marquette, and Vidal). Leaf susceptibility was highest when inoculated at 1 to 2 days old, and inoculated leaves were moderately susceptible at 3 to 6 days old and almost resistant when older than 6 days. The influence of leaf age on anthracnose relative severity was adequately described by an exponential decay model. The susceptibility of the inflorescences was high when inoculated from their initiation to the full flowering (50% fall of the caps), and the inflorescences/flowers were moderately susceptible until veraison, after which the berries were practically resistant. The flower/berry susceptibility as a function of degree-days accumulated since 1 April was modeled using a sigmoid model. Based on this model, 50% disease incidence is reached when 656, 543, and 550 degree days are accumulated for the cultivars Vandal-Cliche, Marquette, and Vidal, respectively. These results suggest that the risk of anthracnose development is high from bud-break to fruit set, and on newly emerged leaves either early in the season or following pruning. More knowledge on anthracnose epidemiology is needed, but these results could be used to improve timing of fungicide applications and pruning activities.

Based on marine benthic diatoms collected at Potter Cove (Western Antarctic Peninsula), we present a detailed study, using both light and electron microscopy, of the morphology of Campyloneis frenguelliae, a species recently transferred from Cocconeis. Ultrastructural observations revealed a combination of new and unusual features within the Achnanthales: a raphe valve with different areolation patterns, a hollow valvocopula with no apparent fimbriae, and a sternum valve externally ornamented by an embossed structure, areolae occluded by hymenes mostly supported by one short peg, and multiple marginal pores that open internally into shallow valve depressions. In view of these unique set of features, we describe Australoneis gen. nov. to include Australoneis frenguelliae comb. nov. and designate a sample from Potter Cove as epitype of the latter. The main characteristics of the new genus are compared to those of its closely allied Cocconeis, Campyloneis, Xenococconeis and Amphicocconeis.

The branchial attaching, fish parasitic genus Mothocya Costa, in Hope, 1851 is recorded for the first time from Nigerian brackish waters on Monodactylus sebae (Perciformes: Monodactylidae). Mothocya andonin. sp. is characterised by a trapezoid-shaped cephalon with a subtruncate rostrum; pereonite 1 anterolateral angles only reaching to the posterior margin of the eyes; particularly large and wide coxa 7 extending to pleonite 4; pereopod 7 with a narrow basis; broadly rounded pleotelson; and uropods extending to the pleotelson posterior margin, with subequal rami. Mothocya powellin. sp. is characterised by its small size (7 mm); a posteriorly ovoid body shape; cephalon anterior margin truncate; a short pleon with pleotelson lateral margins converging to a narrowly rounded apex; pleonite lateral margins overlapped by pereonite 7; slender uropods that extend to the posterior margin of the pleotelson with the uropodal exopod almost double the length of the endopod., Graphical abstract Image 1, Highlights • Mothocya andoni n. sp. and Mothocya powelli n. sp. are described. • First records of Mothocya from Nigeria. • Full descriptions and identifying characteristics for the two new species are provided. • Increased knowledge on the understudied eastern South Atlantic.

In Europe wildlife animals such as the red fox (Vulpes vulpes) are considered the main reservoir for Angiostrongylus vasorum as well as a potential threat for domestic dog infection. Though this parasite is endemic in fox populations, data on A. vasorum infection in wolves (Canis lupus italicus) are still scant, having only recently been described in Northwestern Spain, in Italy, in Croatia and in Slovakia. Based on the rising number of cases of canine lungworm infection in Central Italy (Abruzzo region), the aim of the present study was to investigate the infection by A. vasorum in fox and wolf populations sharing the same geographical area of dogs. From October 2008 to November 2019, A. vasorum specimens were collected, through routine post-mortem examination, from 56 carcasses (44 foxes and 12 wolves). Adult parasites were searched for in the right side of the heart and in pulmonary artery of all carcasses. First stage of larvae (L1) was searched in faeces using the Baermann technique and in lungs by tissue impressions. Overall, 230 adult specimens were collected and identified on a morphological basis. To confirm the morphological identification, 4 adult specimens (n = 3 from fox, n = 1 from wolf) were molecularly identified as A. vasorum by amplification of partial fragment of nuclear 18S rRNA (~1700 bp) genes. The anatomo-pathological and parasitological examinations indicated the presence of A. vasorum in 33 foxes (75%) and in 8 wolves (66.7%). The level of prevalence of infested wolves was higher than the previous one reported in other European countries. Interestingly, the prevalence of infection in foxes herein recorded was higher than that described in dogs (8.9%) living in the same geographical area. This result may confirm the hypothesis that the spread of canine angiostrongylosis is linked to fox populations infection., Graphical abstract Image 1, Highlights • Angiostrongylus vasorum was found in dead foxes and wolves in Chieti province, Italy. • The prevalence in foxes was higher than that described in dogs. • The prevalence in wolves was higher than the previous one reported in European countries. • The spread of canine angiostrongylosis is linked to fox populations infection. • The province of Chieti appears to be an enzootic focus for canine angiostrongylosis.

The American pumpkinseed Lepomis gibbosus, was introduced to Europe more than one hundred years ago. Currently it is a common fish in European freshwaters but relatively few specific parasites infect this fish in this new habitat. In Europe only a single species, Myxobolus dechtiari seems to represent the American myxosporean fauna of centrarchid fishes. M. dechtiari was found in both Portugal and Hungary. This species forms plasmodia with elongated shape inside the cartilaginous rays of gill filaments. In the advanced stage of infection, after disruption of plasmodia, small groups of myxospores remain enclosed in the cartilaginous gill rays causing distortions in the filaments. Myxospores were ellipsoidal in frontal view and lemon-shape in sutural, length 12.5 ± 0.46 (12–13.4) μm, width 10 ± 0.37 (9.6–10.4) μm, and thickness 7.4 ± 0.37 (7–8) μm; the polar capsules were pyriform, equal in size, length 5.6 ± 0.21 (5.3–6) μm, width 3.2 ± 0.16 (3–3.6) μm; Seven to eight polar tube coils were arranged perpendicularly to the capsule length. There was a small, round, 0.4 ± 0.1 (0.3–05) (N = 50) intercapsular appendix in the spores. The small subunit ribosomal DNA (ssrDNA) of M. dechtiari differed from other myxozoans sequenced to date. Phylogenetic analysis of the ssrDNA gene sequence placed this species in a clade including actinospores and Myxobolus species: Raabeia type1, Triactinomyxon sp., and Myxobolus osburni infecting the same host fish. The focus of our study was to prove that the pumpkinseed, a fish originated from North-America introduced one of its myxosporean parasite to Europe. Emphasis was put on to demonstrate the unique feature of this parasite causing infection in the cartilaginous gill rays., Graphical abstract Image 1, Highlights • American pumpkinseed fish were introduced to Europe as an ornamental fish. • Relatively few specific parasites are reported from the pumpkinseed in Europe. • The myxosporean Myxobolus dechtiari described originally in Canada infects the introduced host both in Hungary and Portugal. • SsrDNA sequences of M. dechtiari from pumpkinseed fish is provided.

A single fungal pathogen was consistently isolated from symptomatic wood of olive trees (Olea europaea) displaying branch and trunk cankers in superhigh-density orchards in the Sacramento and San Joaquin Valleys of California. Morphological characters of the pathogen included two distinct types of conidia (thick-walled, dark brown, and globose and thin-walled, hyaline, and oblong to ellipsoid) and three types of phialides, indicating a pleurostoma-like fungus. Phylogenetic results of four nuclear loci including the internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2) and partial sequences of the actin, beta-tubulin, and translation elongation factor 1-α genes confirmed the isolates as Pleurostoma richardsiae. Pathogenicity trials conducted in the field involving 2- to 3-year-old branches of three widely planted oil olive cultivars (Arbequina, Arbosana, and Koroneiki) satisfied Koch’s postulates and confirmed the pathogenic nature of this species to cause the decline of olive trees in California. All three cultivars were equally susceptible to Pl. richardsiae, indicating no detectable resistance to the pathogen. Additional isolations from symptomatic hosts including almond, peach, pistachio, and plum, also confirmed this species, suggesting that Pl. richardsiae is widespread in agricultural systems and should be considered an emerging pathogen of fruit and nut crops in California.

Anthracnose is a destructive disease of alfalfa (Medicago sativa) that causes severe yield losses. Biological control can be an effective and eco-friendly approach to control this alfalfa disease. In the present study, Bacillus amyloliquefaciens LYZ69, previously isolated from healthy alfalfa roots, showed a strong in vitro antifungal activity against Colletotrichum truncatum, an important causal agent of anthracnose of alfalfa. The strain LYZ69 protected alfalfa plants (biocontrol efficacy of 82.59%) from anthracnose under greenhouse conditions. The cell-free culture (CFC) of LYZ69 (20%, vol/vol) caused 60 and 100% inhibition of mycelial growth and conidial germination, respectively. High-performance liquid chromatography tandem mass spectrometry separated and identified cyclic lipopeptides (LPs) such as bacillomycin D and fengycin in the CFC of LYZ69. Light microscopy and scanning electron microscopy revealed that the mixture of cyclic LPs produced by LYZ69 caused drastic changes in mycelial morphology. Fluorescence microscopy showed that the LPs induced reactive oxygen species accumulation and caused apoptosis-like cell death in C. truncatum hyphae. In summary, our findings provide evidence to support B. amyloliquefaciens LYZ69 as a promising candidate for the biological control of anthracnose in alfalfa.

Borrelia burgdorferi sensu lato (s.l.) is the most common pathogen of medical significance transmitted by ticks of the family Ixodidae in Belarus. Human infection with B. burgdorferi causes Lyme borreliosis, most commonly referred to as Lyme disease. Currently, 20 species of Lyme disease-associated Borrelia and more than 20 relapsing fever-associated Borrelia species have been identified. These etiologic agents belong to the genus Borrelia in the family Spirochaetaceae. Genetically characterized isolates with specific sequences have proven that these pathogens are endemically transmitted in many European and Asian countries. In addition, joinpoint regression analysis is often applied to characterize infection trends over time and to identify the time point(s) at which the trend significantly changes. In this epidemiological investigation, joinpoint analysis was applied to investigate the temporal trend of B. burgdorferi s.l. infections in 4070 ticks collected between April and October 2012-2019. Detection of Borrelia species in ticks is an important tool to determine temporal and geographic distribution and abundance, and to predict the risk of Lyme disease to people in different regions. Our data provide a basis for further studies to determine the distribution and abundance of B. burgdorferi s.l. species in Belarus.

Background: New risk areas for tick-borne encephalitis (TBE) are emerging and the spread of disease and vaccine coverage is unclear in Sweden. We wanted to study the prevalence and levels of TBE-virus (TBEV) antibodies in southern Sweden, and to investigate whether there were individuals with undiagnosed TBE. Materials and Methods: Two cohorts of sera were collected: One group of anonymous individuals in rural areas (AIRA) in Skane and one group of volunteers who often got tick-bites (tick-bitten individuals [TBI]). An enzyme-linked immunosorbent assay for TBEV IgM and IgG was performed, as well as a TBEV neutralization test (NT) in selected individuals. Results: In the AIRA group, there was an IgG seropositivity of 5.3%. There were individuals with high antibody levels both in areas previously considered as risk areas (Bromolla and Knislinge), as well as in another area (Tyringe). In the TBI group, 45% of the individuals were vaccinated according to the questionnaires and IgG seropositivity was 28%. A lower seroprevalence and levels of antibodies were seen in the middle-aged group (50-69 years) compared with younger or elderly study participants. A positive NT revealed several individuals with suspected undiagnosed episodes of TBE. Conclusion: Subclinical or misdiagnosed cases have probably occurred in Skane. Middle-aged individuals had lower levels of IgG, which could indicate either less tick exposure or a lower vaccine response. Less than half of the TBI were vaccinated, an indication that more information about the disease and vaccine might be needed. We conclude that the study motivates an increased awareness of TBEV in the region.

Japanese rock ptarmigans, Lagopus muta japonica, are classified as an endangered species in Japan and are found only in the Japanese Alps. The number of birds has decreased in the last half century and cage protection projects have been undertaken as in situ conservation strategies (one of the projects for the recovery plan of Japanese rock ptarmigan) in the mountains. During the period with cage protections, some chicks died and two Eimeria spp., E. uekii and E. raichoi, were identified in the chicks. Here, we examined the soil within the cages and in the surrounding environment to assess potential sources of infection between July to August 2020. We found high numbers of oocysts in the cages, especially at the back sides where the ptarmigan family frequently congregated, but soils in other areas outside the cages were less contaminated or not contaminated at all. The time required for more than 50% of the oocysts to sporulate at 15, 20 and 25 °C for E. uekii was 20, 11, and 5 h, respectively, and 72, 48 and 18 h, respectively, for E. raichoi. Our results cast some doubt that coprophagia by chicks is the source of infection because chicks consumed fresh cecal feces (approximately within 1 h) as far as we know, and instead, the protected chicks might be directly or indirectly infected by oocysts in soils or the environment., Graphical abstract Image 1, Highlights • Cage protection is effective for protecting chicks of Japanese rock ptarmigans. • Soils at the back sides in the cages were highly contaminated with Eimeria spp. • E. uekii can rapidly be sporulated at 15 °C in timber regions. • Protected chicks might be infected by oocysts in soils or the environment.

Fascioloidosis is an allochthonous parasitic disease in Europe caused by the digenean trematode Fascioloides magna. The final hosts of F. magna in Europe are defined as definitive, aberrant and dead-end. Roe deer are aberrant hosts in which juvenile flukes permanently migrate through the liver parenchyma. Failure in pseudocysts formation leads to the death of both the host and the parasite. In this paper we present gross and histological findings of F. magna infection in 34 roe deer. The special emphasis is on the pseudocyst formation accompanied with new fluke's migratory channels observed in 7 animals, suggesting reinfection and prolonged survival of roe deer. No F. magna eggs were recovered from the faeces of the infected animals. These findings indicate that pseudocyst formation is essential for roe deer survival, but also point out the potential beginning of adaptation processes in roe deer, altering otherwise acute and fatal disease into a chronic one.

Diseases caused by fungi can affect the quality and yield of the leaves of tea [Camellia sinensis (L.) Kuntze]. At present, the availability of highly effective and safe fungicides for controlling tea plants remains limited. The objectives of this study were to identify novel compounds with antifungal activities and to determine their molecular mechanisms. A series of sulfone compounds containing 1,3,4-oxadiazole were evaluated in China for their antifungal activities against several pathogens causing foliar diseases and high production losses. Transcriptomics and bioinformatics were used to analyze the differentially expressed genes of Lasiodiplodia theobromae treated with a representative compound, jiahuangxianjunzuo (JHXJZ). Moreover, the effects of JHXJZ on ergosterol content, membrane permeability, cell structure, and seven key genes involved in the ergosterol biosynthetic pathway were investigated. JHXJZ had a strong antifungal activity against L. theobromae in vitro, with an effective concentration giving 50% inhibition of 3.54 ± 0.55 μg/ml, and its curative efficacies on detached tea leaves reached 41.78% at 100 μg/ml. JHXJZ upregulated 899 genes (P < 0.05) and downregulated 1,185 genes (P < 0.05) in L. theobromae. These genes were found to be associated with carbohydrate metabolic processes, which are closely related to steroid biosynthesis in the Kyoto Encyclopedia of Genes and Genomes pathways. Because JHXJZ regulates the key genes of sterol biosynthesis, it decreased the ergosterol content, increased cell-membrane permeability, changed the cellular structure, enhanced the roughness of the surface of the hyphae, and resulted in degradation of the hyphal nuclei and necrosis of the hyphal cytoplasm. Our study demonstrates that JHXJZ is a fungicide with a novel mechanism of action that differs from that of triazole fungicides. JHXJZ has potential for applications in controlling tea plant diseases. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Malaria vectors are supposedly uncommon in urban areas owing to the lack of suitable breeding sites for their development. However, the maintenance in urban areas of traditional rural practices alo...

Within the genus Trichinella, Trichinella pseudospiralis is the only recognized non-encapsulated species known to infect mammals and birds. In October 2020, larvae recovered from muscle tissues of a wolf (Canis lupus italicus) originating from Molise Region, Central Italy, were molecularly confirmed as those of Trichinella britovi and T. pseudospiralis. This is the first detection of T. pseudospiralis from a wolf. In Italy, this zoonotic nematode was detected in a red fox (Vulpes vulpes), three birds (Strix aluco, Athene noctua, Milvus milvus) and five wild boars (Sus scrofa), and was also identified as the etiological agent of a human outbreak of trichinellosis in 2015. Since T. pseudospiralis is rarely reported from carnivore mammals in comparison to the encapsulated species frequently detected in these hosts, this finding opens the question of the role of carnivores as reservoirs for this parasite., Graphical abstract Image 1, Highlights • First report of Trichinella pseudospiralis infection in a wolf. • T. pseudospiralis is rarely documented in carnivorous mammals. • T. pseudospiralis and T. britovi mixed infection in a wolf.

Thelohanellus nikolskii, Achmerov, 1955 is a well-known myxozoan parasite of the common carp (Cyprinus carpio L.). Infection regularly manifests in numerous macroscopic cysts on the fins of two to three month-old pond-cultured carp fingerlings in July and August. However, a Thelohanellus infection is also common on the scales of two to three year-old common carp in ponds and natural waters in May and June. Based on myxospore morphology and tissue specificity, infection at both sites seems to be caused by the same species, namely T. nikolskii. This presumption was tested with molecular biological methods: SSU rDNA sequences of myxospores from fins of fingerlings and scales of older common carp were analysed and compared with each other and with related species available in GenBank. Sequence data revealed that the spores from the fins and scales represent the same species, T. nikolskii. Our study revealed a dichotomy in both infection site and time in T. nikolskii-infections: the fins of young carp are infected in Summer and Autumn, whereas the scales of older carp are infected in Spring. Myxosporean development of the species is well studied, little is known, however about the actinosporean stage of T. nikolskii. A previous experimental study suggests that aurantiactinomyxon actinospores of this species develop in Tubifex tubifex, Muller, 1774. The description included spore morphology but no genetic sequence data (Szekely et al., 1998). We examined >9000 oligochaetes from Lake Balaton and Kis-Balaton Water Reservoire searching for the intraoligochaete developmental stage of myxozoans. Five oligochaete species were examined, Isochaetides michaelseni Lastochin, 1936, Branchiura sowerbyi Beddard, 1892, Nais sp., Muller, 1774, Dero sp. Muller, 1774 and Aelosoma sp. Ehrenberg, 1828. Morphometrics and SSU rDNA sequences were obtained for the released actinospores. Among them, from a single Nais sp., the sequence of an aurantiactinomyxon isolate corresponded to the myxospore sequences of T. nikolskii.

Shelters in Jordan accommodate a huge number of dogs, which are rescued as stray dogs from different cities of the country, but their health receives almost no attention. The aim of this study was to examine tick infestation as well as tick-borne protozoa and bacteria of 80 randomly sampled dogs in two Jordanian shelters. Ticks identified as Rhipicephalus sanguineus sensu lato were found on 14 out of 27 animals in a shelter. No ticks were found on dogs in the other shelter. A total of 42 (52.5% [95% confidence interval: 41.7-63.1]) dogs were infected with one or two pathogens. The DNA of three protozoal (Hepatozoon canis, Babesia vogeli, and Babesia negevi) and two bacterial (Anaplasma platys and Candidatus Bartonella merieuxii) species were detected in the blood samples. To the best of the authors' knowledge, except for H. canis, these species are reported for the first time from Jordan.

Hyaloklossia Labbé ,1896 (Alveolata: Apicomplexa) is a monotypic genus of renal coccidia found in anurans, particularly in the edible frog Pelophylax kl. esculentus (Amphibia: Anura: Ranidae), distributed in different parts of Europe. Here we propose a new Hyaloklossia species from the Tokyo daruma pond frog, Pelophylax porosus porosus. The coccidium detected in the renal tissue of P. p. porosus shared some morphological characteristics with the type species, Hyaloklossia lieberkuehni (Labbé, 1894), reported from P. kl. esculentus. However, in addition to size differences in several oocyst and sporocyst features between these parasites, phylogenetic analysis of gene fragments from two nuclear ribosomal loci and the mitochondrial cytochrome c oxidase subunit 1, exposed distinct genetic differences between H. lieberkuehni and our new species. Although our analysis validated the monophyly of Hyaloklossia with some members of the Toxoplasmatinae Biocca, 1957, Cystoisosporinae Frenkel et al., 1987, and Eumonosporinae Chou et al., 2021 (Sarcocystidae Poche, 1913), comparison of genetic differences between Hyaloklossia species from P. p. porosus and H. lieberkuehni revealed the presence of a greater number of polymorphisms than that observed when comparing inter-species (Heydornia spp., Besnoisita spp.) or inter-genus (Toxoplasma vs. Neospora, Neospora vs. Hammondia, and Neospora vs. Heydornia) variabilities among members of the Sarcocystidae. This indicates that Hyaloklossia, as re-erected and defined by Modrý et al. (2001, Int. J. Syst. Evol. Microbiol. 51, 767–772), with its homoxenous life cycle, requires placement in its own subfamily. Thus, we propose a new subfamily, Hyaloklossiinae n. subfam., to accommodate two species, H. lieberkuehni from Europe and Hyaloklossia kasumienesis n. sp. which we describe here from P. p. porosus in Japan., Graphical abstract Image 1, Highlights • Hyaloklossia was detected from the kidney of Pelophylax porosus porosus (Amphibia: Anura). • The species was described as Hyaloklossia kasumiensis n. sp. • This is a new host record for Hyaloklossia and also the first report in Asia. • Hyaloklossiinae, was proposed as a new subfamily in the Sarcocystidae.

Rice bacterial blight is a devastating bacterial disease threatening rice yield all over the world and Xanthomonas oryzae pv. oryzae is traditionally believed to be the pathogen. In recent years, we have received diseased rice samples with symptoms of blighted leaves from Sichuan and Guangdong provinces, China. Pathogen isolation and classification identified two different enterobacteria as the causal agents, namely Enterobacter asburiae and Pantoea ananatis. Among them, E. asburiae was isolated from samples of both provinces, and P. ananatis was only isolated from the Sichuan samples. Different from rice foot rot pathogen Dickeya zeae EC1 and rice bacterial blight pathogen X. oryzae pv. oryzae PXO99A, strains SC1, RG1, and SC7 produced rare cell wall degrading enzymes (CWDEs) but more extrapolysaccharides (EPS). E. asburiae strains SC1 and RG1 produced bacteriostatic substances while P. ananatis strain SC7 produced none. Pathogenicity tests indicated that all of them infected monocotyledonous rice and banana seedlings, but not dicotyledonous potato, radish, or cabbage. Moreover, strain RG1 was most virulent, while strains SC1 and SC7 were similarly virulent on rice leaves, even though strain SC1 propagated significantly faster in rice leaf tissues than strain SC7. This study firstly discovered E. asburiae as a new pathogen of rice bacterial blight, and in some cases, P. ananatis could be a companion pathogen. Analysis on production of virulence factors suggested that both pathogens probably employ a different mechanism to infect hosts other than using cell wall degrading enzymes to break through host cell walls.

Ticks are important vectors that facilitate the transmission of a broad range of micropathogens to vertebrates, including humans. Because of their role in disease transmission, it has become increasingly important to identify and characterize the micropathogen profiles of tick populations. The objective of the present study was to survey the micropathogens of ticks by third-generation metagenomic sequencing using the PacBio Sequel platform. Approximately 46.481 Gbp of raw micropathogen sequence data were obtained from samples from four different regions of Heilongjiang Province, China. The clean consensus sequences were compared with host sequences and filtered at 90% similarity. Most of the identified genomes represent previously unsequenced strains. The draft genomes contain an average of 397,746 proteins predicted to be associated with micropathogens, over 30% of which do not have an adequate match in public databases. In these data, Anaplasma phagocytophilum and Coxiella burnetii were detected in all samples, while Borrelia burgdorferi was detected only in Ixodes persulcatus ticks from G1 samples. Viruses are a key component of micropathogen populations. In the present study, Simian foamy virus, Pustyn virus and Crimean-Congo haemorrhagic fever orthonairovirus were detected in different samples, and more than 10–30% of the viral community in all samples comprised unknown viruses. Deep metagenomic shotgun sequencing has emerged as a powerful tool to investigate the composition and function of complex microbial communities. Thus, our dataset substantially improves the coverage of tick micropathogen genomes in public databases and represents a valuable resource for micropathogen discovery and for studies of tick-borne diseases., Graphical abstract Image 1, Highlights • The microbial communities from ticks were analysed by third-generation metagenomic sequencing using the PacBio Sequel platform. • In these data, Anaplasma phagocytophilum and Coxiella burnetii were found in four groups, and Borrelia burgdorferi was detected only in Ixodes persulcatus ticks from G1 samples. Viruses are a key component of the composition of micropathogens. • The third-generation metagenomic sequencing is far superior to second-generation sequencing in genome sequence integrity, and the similarity of the sequences obtained via third-generation metagenomic sequencing for discrimination is unmatched by other sequencing methods. • Thus, our dataset substantially improves the coverage of tick micropathogen genomes in public databases and represents a valuable resource for micropathogen discovery and for studies of tick-borne diseases.

West Nile virus (WNV) is a mosquito-borne flavivirus that can cause severe neurological disease in humans, for which there is no treatment or vaccine. From 2009 to 2018, California has reported more human disease cases than any other state in the United States. We sought to identify smaller geographic areas within the 10 California counties with the highest number of WNV cases that accounted for disproportionately large numbers of human cases from 2009 to 2018. Eleven areas, consisting of groups of high-burden ZIP codes, were identified in nine counties within southern California and California's Central Valley. Despite containing only 2% of California's area and 17% of the state's population, these high-burden ZIP codes accounted for 44% of WNV cases reported and had a mean annual incidence that was 2.4 times the annual state incidence. Focusing mosquito control and public education efforts in these areas would lower WNV disease burden.

The Leadbeater's possum (Gymnobelideus leadbeateri) is a critically endangered marsupial in south-eastern Australia. Among other conservation efforts, free-ranging animals in the two remaining geographically separate populations (highland and lowland) have been extensively studied; however, little is known about their health and mortality. Although some wild populations are frequently monitored, cadavers are rarely recovered for post mortem examination. In June 2019, a recently deceased, wild, adult male lowland Leadbeater's possum was collected from a nest box and a comprehensive post mortem examination was conducted. Microfilariae of a filarioid nematode were observed in testes, liver, lung and skin samples in tissue impression smears and upon histopathological examination. No gross or histological changes were seen associated with the parasites, except for a focal area of tissue damage in the skin, suggesting that the possum is a natural host. Using a PCR-coupled sequencing method the filarioid was identified as a species of Breinlia. Species of Breinlia occur in other Australian marsupials and rodents.

Diet is an important factor in understanding bat ecology and conservation. This study assessed seasonal prey availability and diet composition of the Asiatic Lesser Yellow House Bat Scotophilus kuhlii in various districts of Uttar Pradesh between January 2016 to December 2018. Fecal and insect samples were collected seasonally using sweep nets between 1800 and 1900 h. From each location 20 fecal pellets were selected for analysis and searched for taxonomically recognizable remnants. The analysis revealed that S. kuhlii fed on Coleoptera, Diptera, Hymenoptera, Isoptera, Orthoptera, Odonata, Blattodae, Lepidoptera, and Hemiptera, identified from legs, antennae and wings/elytra in fecal pellets. Seasonal variation in the presence of isolated insect remnants and insect abundance at foraging grounds was observed. Thus S. kuhlii is a voracious feeder and plays an important role as a pest control agent.

In this study, the pollen morphology of all Ferula species distributed throughout the country of Turkey was studied with light and scanning electron microscopy for the first time. The aim is to identify the pollen morphological characteristics of 23 Ferula species. The pollen is radially symmetrical, isopolar and tricolporate in all examined species. Pollen grains are prolate and perprolate with the polar axis ranging from 22.28 to 40.47 µm and the equatorial axis from 13.70 to 18.73 µm. Their polar shapes are triangular, triangular to subtriangular and circular to subcircular. Several types of exine ornamentations have been observed on pollen through the use of scanning electron microscopy. The dendrogram constructed by using Average Linkage of the examined data revealed two main groups. It was determined that some pollen characteristics are more useful for classification than others. In particular, P, E, the ratio of P/E (pollen shape) and ornamentation in the polar and equatorial views are the most valuable variables for discrimination the Ferula species.

The paper describes the GUSMP (North Sea Route Directorate) expert council session on the design of a powerful icebreaker of 18000–24000 HP held on June 15, 1935 in Leningrad, based on the archive documents. Analysis of the session materials sheds light on the state-of-the-art of icebreaker building in the mid-1930s. The standpoint of academician A.N. Krylov as well as GUSMP administration is particularly interesting.

The objective of this study was to evaluate the effectiveness of the main anthelmintics used for the control of trematodes in cattle in an endemic area in the humid Mexican tropics. A diagnostic study was carried out in nine cattle farms to determine the prevalence of flukes through faecal examination. Only three farms with more than 20 cows positive to trematodes were chosen to determine the effectiveness of commercial anthelmintics (triclabendazole, TCBZ; ivermectin + closantel, (IVM + CLOS); IVM + clorsulon, (CLORS); nitroxynil, NITROX). The prevalence of Fasciola hepatica was 27.1% and 29.6% of paramphistomes. The faecal egg count of trematodes ranged from 0.0 to 12.2 eggs per gram of faeces. The highest effectiveness against F. hepatica was 96.7%, and 92.7% against paramphistomes. NITROX was the most effective in the control of trematodes, while other products, such as IVM + CLORS and TCBZ obtained values lower than 90%, which puts sustainable trematode control at risk. The presence of trematodes was observed on most farms, although the prevalence per herd was highly variable, which indicates that the trematodes F. hepatica and paramphistomes are endemic to the region and a suitable management programme is suggested to control infections caused by these parasites.

Neospora caninum (Family: Sarcocystidae) is an obligate intracellular protozoan. It is one of the most critical abortifacients in ruminants. The seroprevalence of antibodies against N. caninum and its risk factors was investigated among 430 sheep from four North Egyptian governorates, Alexandria, Gharbia, Menofia, and Qalyubia, during the period from 2017 to 2018. Generally, the overall prevalence rate of N. caninum among sheep was 8.6%. The logistic regression analysis for the obtained data revealed that N. caninum increased significantly with age (OR = 2.4, 95% CI: 8.4–18.7) of the ewe (OR = 3.3, 95% CI: 7.6–14.9), particularly among sheep in contact with dogs (OR = 4.9, 95% CI: 7.5–14.3). Besides, locality, season, and pregnancy status of examined sheep had no significant effect on the appearance of N. caninum infection. the present findings confirm the presence of N. caninum among sheep in Egypt which probably play a role in reproductive failure in sheep. Therefore, sanitary measures and monitoring of the infection should be implemented to reduce the spreading of the infection.

Introduction The aim of the study was to carry out epizootic assessment of male roe deer to detect the presence of Cephenemyia stimulator larvae and determine the influence of the parasite on the carcass and antler weight in animals living in different habitats. Material and Methods The investigations were based on post-mortem analysis of Cephenemyia stimulator infestations of the nasal passages and throat of 177 male roe deer culled between May 11 and September 30, 2020 in hunting districts of the Lublin region in Poland. The individual quality of the animals was assessed by weighing the gutted carcasses after cooling, and the antlers were weighed after dissection and their total weight was determined. Results The parasite prevalence ranged from18 to 48% according to habitat type, with a mean value of 33%. The highest prevalence was detected in bucks living in grassland ecosystems. The presence of the parasites exerted influence on the individual condition of the animals, which was reflected in reduced carcass and antler weight. The differences were found both in young individuals and in somatically fully developed animals, but they were not statistically significant in all age groups. Conclusion Although the parasitic infestations impaired the individual condition traits, their parametric values in most cases were not lower than those reported from other regions of Poland. Despite the Cephenemyia stimulator infestation, male roe deer from the Lublin region are characterised by high carcass and antler weight.