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BackgroundWheat is the main cultivated crop and the highest cereal used up by humans in Iraq. Two Fusarium species causing Fusarium head blight (FHB) disease on wheat has been detected only in the middle of Iraq, and Fusarium crown rot (FCR) disease has been detected in several provinces, but it was limited to only three species. This study was conducted to deepen and enlarge our current understanding of the causal agents of these symptoms observed in the wheat fields and to isolate, purify, and identify (molecularly) the most common soil fungi occurring in the rhizosphere of seedlings showing, root, crown, stem, and head at symptom occurrence.ResultsEighty-eight Fusarium isolates were isolated from 7 wheat cultivars planted in 14 fields in the south of Iraq, Basra province, by using polymerase chain reaction (PCR) technique. Based on molecular identification methods as well as the analysis of phylogenetic trees, the results of this study have verified that the main species belonging to the genus Fusarium causing FHB and FCR diseases on wheat in the studied areas are F. pseudograminearum, F. graminearum, F. equiseti, F. culmorum, F. solani, F. avenaceum, F. chlamydosporum, F. cerealis, and F. nygamai. The first 3 species have been identified previously in several Iraqi studies on wheat and barley. This is the first record of the last 6 Fusarium species from the wheat crop in Iraq as causative pathogens of FHB and FCR. Individual phylogenetic trees of 71 Fusarium isolates reconstructed based on the internal transcribed spacer (ITS) sequences showed that all isolates belonging to each of the 9 Fusarium species form monophyletic groups with identical isolates of the same species. Collective phylogenetic tree of the remaining 17 Fusarium isolates reconstructed based on translation elongation factor 1 alpha (TEF1-α) gene sequences showed that all isolates belonging to each of the 4 Fusarium species forms monophyletic groups with identical isolates of the same species.ConclusionIraqi wheat cropping system is seriously threatened by FHB and FCR diseases. Other Iraqi provinces should be inspected to understand the distribution of both diseases.

This study was aimed to the detection of Nosema in European honeybees at Kanchanaburi Province and Identify species of Nosema by Polymerase Chain Reaction technique. The ventriculus of bees was individually checked to verify the presence of Nosema spores under light microscope. The number of spores per bee were quantify on a haemocytometer for infectivity. It was studied for three periods of the year. The first period was studied between October 2015 to January 2016, the second period from February to May 2016 and the third period from June to September 2016. The results showed that the highest infection rate in June-September 2019 was 100 and the lowest infection rate was 40 in October 2015 and January 2019. The average number of Nosema spores infection level in an individual was infectivity. The highest infectivity was 2.47x106 spores/bee in February to May 2016 and the lowest infectivity was 0.04x106 spores/bee in October 2015 to January 2016. Nosema ceranae was found in this study by Polymerase Chain Reaction technique.

Bletilla striata (Thunb.) Rchb. f. (Orchidaceae family, known as Baiji in Chinese) is a perennial herb and has been traditionally used for hemostasis and detumescence in China. In April of 2020, a leaf spot disease on B. striata was observed in plant nurseries (∼0.2 h) in Guilin, Guangxi Province, China. Approximately 20% of the plants were symptomatic, of which 150 plants were randomly selected for investigation. Initial symptoms include the appearance of small, circular or irregular light brown spots, randomly scattered on the edges and surfaces of the leaves, which progressively expand into large, suborbicular or irregular-shaped dark brown, necrotic areas. At the severe stage, the lesions coalesced into large necrotic areas and ultimately resulted in leaf abscission. To isolate the pathogen, three representative plants exhibiting symptoms were collected from the nurseries. Leaf tissues (5 × 5 mm) were cut from the margin of necrotic lesions (n = 18), surface-disinfected in 1% sodium hypochlorite (NaOCl) solution for 2 min, then rinsed three times in sterile water before isolation. The tissues were plated on potato dextrose agar (PDA) medium, and incubated at 28°C (12-h photoperiod) for 3 days. Hyphal tips from recently germinated spores were transferred to PDA to obtain pure cultures. Nine fungal isolates with similar morphological characteristics were obtained. Three single-spore isolates, BJ23.1, BJ55.1, and BJ91.3, were subjected to further morphological and molecular characterisation. Colonies on PDA plates were villose, had a dense growth of aerial mycelia and appeared white (1A1) to yellowish white (3A2). Macroconidia were smooth, hyaline, straight to slightly curved, usually contained three or five septa, and measuring 23.3 to 42.1 × 3.0 to 6.2 μm (mean ± SD: 31.2 ± 5.1 × 4.2 ± 0.6 μm, n = 50). Microconidia were generally cylindrical, straight to slightly curved, aseptate, and measuring 7.2 to 18.8 × 2.5 to 4.3 μm (mean ± SD: 12.1 ± 2.8 × 3.3 ± 0.5 μm, n = 62). Morphological characteristics are similar to those of F. commune (Skovgaard et al. 2003). For molecular identification, the genomic DNA of the isolates BJ23.1, BJ55.1, and BJ91.3 were extracted using the CTAB method (Guo et al. 2000). The internal transcribed spacer (ITS) region of rDNA, partial translation elongation factor-1 alpha (TEF-1α), RNA polymerase second largest subunit (RPB2), and the mitochondrial small subunit rDNA (mtSSU) genes were amplified using primer pairs [ITS1/ITS4 (White et al. 1990), EF-1/EF-2 (O'Donnell et al. 1998), and 5f2/11ar (Liu et al. 1999, Reeb et al. 2004), MS1/MS2 (Li et al. 1994), respectively]. The obtained sequences were deposited in NCBI GenBank under the following accession numbers: ITS (MZ424697 to MZ424699), TEF-1α (MZ513467 to MZ513469), RPB2 (MZ513473 to MZ513475), and mtSSU (MZ513470 to MZ513472). BLAST® analysis of the deposited sequences showed 99 to 100% identity with those of F. commune present in GenBank (Accession numbers: DQ016205, MH582348, MH582181, AF077383). In addition, a phylogenetic analysis using concatenated sequences of ITS, TEF-1α, mtSSU genes showed that BJ23.1, BJ55.1, and BJ91.3 located on the same clade with strains of F. commune. Therefore, based on morphological and molecular characteristics, the isolates were identified as F. commune (Skovgaard et al. 2003, Stewart et al. 2006). Pathogenicity was tested using 1.5-year-old B. striata plants. Healthy leaves on plants were inoculated with 5 × 5 mm mycelial discs of strains BJ23.1, BJ55.1, and BJ91.3 from 3-day-old PDA cultures, each isolate was inoculated onto three plants; three other plants inoculated with sterile PDA discs served as controls. All plants were enclosed in transparent plastic bags and incubated in a greenhouse at 28°C for 14 days (12-h photoperiod). Three days post-inoculation, leaf spot symptoms appeared on the inoculated leaves. No symptoms were detected on control plants. Experiments were replicated three times with similar results. To fulfill Koch's postulates, F. commune was consistently re-isolated from symptomatic tissue and confirmed by morphology and sequencing, whereas no fungus was isolated from the control plants. F. commune has been reported to cause diseases on some plants, including sugarcane (Wang et al. 2018), maize (Xi et al. 2019) and Wax Gourd (Zeng et al. 2020). To our knowledge, this is the first report of F. commune causing leaf spot disease on B. striata in China. Identification of this pathogen provides the information for further studies to develop management strategies to control the disease.

Stratigraphic units during Lower Cretaceous in southern of Iraq were affected largely by tectonic agent. Neo-Tethys ocean occupied the distance between passive margins of Arabian and Iranian plates. South of Iraq represented the continental shelf of passive margin of Arabian plate. This tectonic situation made the sediments in eastern parts of passive margin in south of Iraq effected by marine agents, and the sediments in western parts was effected by continental agents, whereas the middle parts was effected by both agents. As result of that, three sediment regions were formed in south of Iraq. First, marine sediment region that occupy the eastern part of south of Iraq near the Iraq-Iran borders. Second, mixed sediment region that occupy Basra area and extend toward north. Third, continental sediment region that occupy western part of Basra near the western desert. Half graben basins that formed by listric faults in the passive margin caused the local changes in distribution of sediments in south of Basra. These three sediment regions formed all during Lower Cretaceous and maybe extend toward the middle of Iraq and north of Baghdad

A total of 13 samples of domestic cat Felis cattus (Linnaeus, 1758 ) and 9 samples of wild cat Felis chaus furax (de Winton, 1898) of the Felidae Family were trapped and examined to detect the hard ticks. The areas of the collection were: Baghdad, Al-Rashidiya, Tharthar, Nahrawan, AL-Mahmoudiya (middle of Iraq) and AL-Haretha (south of Iraq), Mosul (north of Iraq). The results of the current study revealed that four species belong to two genera of hard ticks: Haemaphysalis sp. (Koch, 1844), Rhipicephalus turanicus (Morel, 1969), Rhipicephalus sanguineus (Neumann, 1904) and Rhipicephalus appendiculatus (Santos, 1955). The rates and the density of infestation were discussed. The current study aimed to clarify the infestation difference between domestic and wild cats with hard ticks (3, 14.88) because domestic cats enjoy human attention, as they live close to him. The current study identified the Rhipicephalus appendiculatus for the first time in Iraq from domestic cat Felis cattus. Keywords: Felidae, Haemaphysalis, Ixodidae, Rhipicephalus, wild cat.

Small ruminants theileriosis are widespread in Iraq andacute infections usually with hight mortality. However, the survived animals suffer from low production of meat and milk. Coinfection with more than Theileria sp. And/or Anaplasmosis could have an impact on the disease severity. The main finding was identifying T. lestoquardi, T. ovis , T. annulata, blood samples of infected sheep with a history of chronic theileriosis (n=48) and with acute clinical theileriosis sign (n=24) were being collected from fields located in Babylon province (middle of Iraq) after chlinical examination and Polymerase chain reaction and real time PCR were performed for detection. Theileria. lestoquardi was the highest of these species within the acute and chronic cases. As well as, the load of this species in acute cases was significantly higher (P<0.01) to that in chronic. However, the load of T. ovis and T. annualta were similar in acute and chronic cases. Importantly, all these cases were coinfected with Anaplasma phagocytophylum. This could be due to the infection of leukocytes meanwhile weakening of the animal's immune system. Also, these parasites transmitted by the same tick-vector. The impact of this finding could help in disease prevention and diagnosis.

The aim of this study was to compare the sensitivity and specificity of polymerase chain reaction (PCR) using two primer pairs and combined with blood culture, immunoglobulin M enzyme-linked immunosorbent assay (IgM ELISA), microscopic agglutination test (MAT) and slide agglutination test (SAT) in the diagnosis of human leptospirosis. We analysed 124 serum samples: 60 from patients with confirmed leptospirosis, 20 from patients with other diseases and 44 from healthy individuals. Analysing the first serum sample collected during the first 3-8 days of disease, the sensitivities of the four tests MAT, IgM ELISA, SAT and PCR were, respectively, 69.0%, 79.3%, 72.4% and 62%. In subsequent samples, those same sensitivities were, respectively, 95.4%, 100%, 100% and 72.7% in samples collected from days 9 to 14 and 88.9%, 88.9%, 77.8% and 44.4% in those collected from days 15 to 42. The most specific method (at 100%) was PCR and the least specific (at 89.1%) was IgM ELISA. Although we found PCR to be less sensitive than the serological tests over the course of the disease, our data indicate that PCR was the most sensitive in those initial serum samples presenting no specific antibodies detectable by any of the serological methods tested. We also recommend that PCR can be used in combination with serological tests as we found that this improves the sensitivity of the diagnosis of leptospirosis in the first phase of the disease (93.1-96.5%).

This thesis describes the isolation and characterisation of novel lectins from Australian indigenous mushroom species and evaluation of their antiproliferative activity against human colon adenocarcinoma (HT29) cell line. In this project, eighteen (18) Australian mushrooms were selected for screening of lectin activity based on published data, availability, distribution, native or exotic, and were collected from the Royal Botanical Garden, Melbourne. The results of the project identified eight (08) mushrooms with lectin activity that led to the isolation and purification of three (03) lectins from three mushroom species. Of these three lectins isolated, two lectins those of, Phlebopus marginatus (PML) and Phaeolus schweinitzii (PSL-2), are novel and one lectin was isolated for the first time from Psathyrella asperospora (PAL). All mushroom species were homogenated with PBS and then were screened for hemagglutination and lectin activity. Hemagglutination activity was evaluated using both neuraminidase treated and untreated rabbit and human A, B, and O erythrocytes. Lectin activity was determined by the ability of various mono- and di-saccharides to inhibit hemagglutination activity. The results of this screening study identified eight (08) mushroom species that showed lectin activity, Agaricus bitorquis, Chlorophyllum brunneum, Coprinus comatus, Cortinarius sp. TWM 1710, Omphalotus nidiformis, Phlebopus marginatus, Psathyrella asperospora and Phaeolus schweinitzii. The remaining ten (10) mushroom species only showed hemagglutination activity devoid of any lectin activity.

Sweet viburnum [Viburnum odoratissimum (L.) Ker Gawl] is an evergreen shrub mainly cultivated along roadsides in urban landscapes and also in parks and residential areas. A foliar disease occurred on about 40% of sweet viburnum plants near Anhui Grand Theatre, Anhui Province of China in June 2019. In early stages of sweet viburnum infection, the symptoms appeared as small brown spots ranged in length from 2 to 3 millimeters on the leaves. The spots developed on the upper, middle, and lower leaves of the plant, however, the upper leaves got more severely affected. As the disease develops, the spots enlarged and became rectangular or oval, brown to dark-brown, and their centers became ashen gray. In later stages of infection, the diseased leaves became wilting. Diseased leaves were surface disinfested and three small sections (2-3 mm2) were cut from the margin of the lesions. Sections were placed in 1.5% NaClO for 2 min, submerged in three changes of sterilized distilled water for 1 min each, placed onto potato dextrose agar (PDA) medium amended with 50 μg/ml of ampicillin and kanamycin, and incubated at 25℃ for 3 days. The mycelium from the leading edge of colonies growing from the tissue was sub-cultured onto a PDA plate for 3 days, followed by spore induction (Simmons 2007) and single spore isolation to obtain a pure culture of the putative pathogen. Colonies of one single spore isolate HF0719 were rounded, grayish white with dense aerial mycelium viewed from above and dark brown viewed from below. On potato carrot agar (PCA) medium, conidiophores were branched or occasionally unbranched. On branched conidiophores, conidia were in dwarf tree-like branched chains of 2-5 conidia. On unbranched conidiophores, conidia were simple or in chains of 2-8 conidia. Conidia were light brown or dark brown, ovoid, ellipsoidal to fusiform, and ranged in size from 7 to 26.5 × 4.5 to 11 μm with an average size of 16 × 7 µm based on 500 spore observations, with one beak and 1-7 transverse, 0-3 longitudinal, and 0-3 oblique septa. Beaks were ranged in (1.5-)2-10(-16) μm long. Based on cultural and morphological characteristics, isolate HF0719 was identified as Alternaria spp. (Simmons 2007). For molecular identification, total genomic DNA was isolated from mycelia collected from 7 day-old colonies of isolate HF0719 using the fungal genomic DNA extraction kit (Solarbio, Beijing, China). Fragments of five genes, including those encoding glyceraldehyde-3-phosphate dehydrogenase (gpd), plasma membrane ATPase, actin, calmodulin, and the Alternaria major allergen (Alt a1) regions of isolate HF0719 were amplified and sequenced using primer pairs gpd1/gpd2 (Berbee et al. 1999), ATPDF1/ATPDR1, ACTDF1/ACTDR1, CALDF1/CALDR1 (Lawrence et al. 2013), and Alt-for/Alt-rev (Hong et al. 2005), respectively. The obtained nucleotide sequences were deposited into GenBank as accession numbers: gpd, MT614365; ATPase, MT614364; actin, MT614363; calmodulin, MN706159; and Alt a1, MN304720. Phylogenetic tree using a maximum likelihood bootstrapping method based on the five-gene combined dataset in the following order: gpd, ATPase, actin, calmodulin, Alt a1 of HF0719 and standard strains representing 120 Alternaria species (Lawrence et al. 2013) was constructed. Isolate HF0719 formed a separate branch. On the basis of morphological characteristics and phylogenetic pattern, isolate HF0719 was identified as Alternaria spp.. A pathogenicity test was performed by rubbing 32 healthy leaves of six 5-year-old sweet viburnum plants with a cotton swab dipped in spore suspension containing 2.6 × 106 spores/ml, following leaf surface disinfection with 70% ethanol in the open field. Sterilized distilled water was used as control. The average air temperature was about 28℃ during the period of pathogenicity test. Eleven days after inoculation, 100% of inoculated leaves showed the leaf spot symptom identical to symptoms observed in the field. Control leaves were symptomless. The experiment was done three times. The re-isolated pathogen from the leaf lesion had the same morphological and molecular characteristics as isolate HF0719, thus satisfying Koch's postulates. The genus Alternaria has been reported to cause leaf spot on sweet viburnum in Florida, USA (Alfieri et al. 1984). To our knowledge, this is the first report of Alternaria spp. causing leaf spot on sweet viburnum in China, a highly valued ornamental plant. Our findings will contribute to monitoring and adopting strategies for manage leaf spot disease on sweet viburnum.

A total of 9 samples of wild cat Felischausfurax (de Winton, 1898) and 13 (11 positives) samples of domestic cat Feliscattus (Linnaeus, 1758) belong to Family Felidae. All cats were trapped and used hair and skin scrapings by forceps and surgical blades. The areas of the collection were: Mosul province (north of Iraq); Baghdad, Al-Rashidiya, Tharthar, Nahrawan, AL-Mahmoudiya (middle of Iraq) and AL-Haretha (south of Iraq). The current study revealed that the sensitive and specific PCR assay allowing rapid and reliable identification of Malasseziarestricta by the fragment size amplified was 500bp in the ITS1 gene in one sample of wild cats. The current study recorded a new strain of Malasseziarestrictathat called AF2013 strain "small subunit ribosomal RNA gene, partial sequence; internal transcribed spacer 1", complete sequence; and 5.8S ribosomal RNA gene, partial sequence. Which was inserted in GenBank: MW376484.1 from wild cat Felischausfurax for the first time in Iraq. Sequencing revealed close matching of the phylogenetic tree to an isolate from Korea (CP030254). The compression was performed using NCBI – the based nucleotides website. Keywords: Dermatitis, Cutaneous microflora, fungi, Genotype, Malasseziarestricta.

The polymerase chain reaction (PCR) was used to identify an isolate of Tomato yellow leaf curl Sardinia virus (TYLCSV) from southwestern Morocco and to detect the members of the Tomato yellow leaf curl virus (TYLCV) complex. Thirty-five tomato samples with typical TYLCV symptoms were collected from infected tomato fields in the Souss-Massa region. PCR was performed with a general primer pair based on the coat protein (Cp) gene of the TYLCV complex, as well as with specific primer pairs for TYLCV and TYLCSV. Of the 35 samples tested, 29 generated a viral DNA product with the general primer pair, 29 samples gave a viral DNA product with the TYLCV-specific primers, and of these, 9 also gave a product with the TYLCSV primer pair; 6 samples did not give any PCR product with either primer pair. The full-length genome of TYLCSV was amplified with overlapping primers at the unique NcoI site in the TYLCSV genome (GenBank accession number X61153). The full-length genome of the TYLCSV isolate from Morocco is 2,777 nucleotides long (accession number AY702650) and is almost identical (97% nucleotide identity) to a TYLCSV isolate from Murcia, Spain (accession number Z25751). A PCR-based diagnostic method was developed to distinguish between TYLCV and TYLCSV in Morocco. To diagnose the TYLCV/TYLCSV complex a general primer pair was designed that anneals to a conserved region of the Cp gene. To diagnose TYLCSV exclusively, two primer pairs were designed to anneal specifically to the replication-associated protein gene (Rep) of TYLCSV from Morocco. To detect TYLCV exclusively, a primer pair previously described to amplify the intergenic region (IR) of TYLCV was used. The PCR primers were tested for their effectiveness using DNA clones of the TYLCSV from Morocco and of the TYLCV from the Dominican Republic. PCR using these primers offers a rapid means to detect the TYLCV complex and to distinguish between the two TYLCV species present in Morocco.

Plum (Prunus salicina Lindl.) is planted in many places in China. In July 2021, plums were found to be covered with unfamiliar yellow mycelia on the plum trees and on the ground in the campus of Northeast Agricultural University in Harbin. The symptoms on the fruit were initially manifested as white mycelium and water damage, then the mycelium turned yellow and the water immersion area expanded rapidly. Finally, the whole fruit is covered by yellow, dense mycelium, so that the flesh becomes soft rot, and fall off, causing about 3~5% economic loss (Fig. 1A, B). The thirty diseased fruits were collected, and the tissue (5 × 5 mm) was cut from the diseased fruits, and then surface sterilized with 75% alcohol for 30 s, soaked in 3% sodium hypochlorite for 5 min, washed with sterile distilled water for 3 times, and inoculated on potato dextrose agar (PDA) and malt extract agar (MEA) at 28 °C. Strains growing around the tissues from samples were subcultured on PDA, and 3 strains (DNMI01, DNMI02 and DNMI03) were obtained using the single-spore isolation method. On PDA, the hyphae were light yellow in the early stage, and became golden yellow in the later stage. The edge of the colony was fan-shaped, and the growth rate was extremely fast for 3 days to reach a full dish (Fig. 1G). On MEA, the mycelium was light yellow at the initial stage, and became grayish black in the later stage. The colony grew irregularly and grew slowly (Fig. 1H). Sporangiospores have two shapes, yellow, aseptate, globose or subglobose 7.7 to 14.1 × 8.5 to 13.9 μm (n=30), oval 2.5 to 6.4 × 6.2 to 12.0 μm (n=30, Fig. 1C). Columellae frequently pyriform, oval, colorless or yellowish, and 31.5 to 53.7 μm × 50.6 to 83.2 μm (n=30, Fig. 1D, E). Sporangia globose or subglobose, yellow, wall echinulate, and 55.3 to 109.1 × 67.9 to 123.5 μm (n=30, Fig. 1F) (Santiago et al. 2013). Acetyl trimethyl ammonium bromide method was used to extract DNA from 3-day-old. The internal transcribed spacer (ITS) and 28S rRNA partial gene sequences were amplified by primers ITS1 / ITS4 (White et al. 1990) and LR0R / LR5 (O'Donnell 1993), respectively. The sequences obtained by polymerase chain reaction (PCR) were sequenced and the sequences were deposited in GenBank with accession numbers ITS, OR136169-OR136171, and LSU, OR136179-OR136181. BLAST analysis of these sequences showed 96.13%-96.48% similarity to Mucor inaequisporus culture CBS255.36 (JN206177.1), 99.28%-99.58% to culture CBS255.36 (MH867301.1), respectively (Walther et al. 2013). Phylogenetic analysis by maximum likelihood method (MEGA7.0) generated based on the ITS, LSU sequences indicated that the strains formed a supported clade to the related M. inaequisporus type sequences. Strains was found to be most closely related to M. inaequisporus and far from other species. Based on morphological and phylogenetic characteristics, strains DNMI01 to DNMI03 was identified as M. inaequisporus (Lee et al. 2020; Ren et al. 2023). According to Koch 's postulate the pathogenicity test was conducted with healthy Prunus salicina fruits. Ten fruits were surface-disinfected with 75% alcohol for 15 s, rinsed with sterile water 3 times, air-dried, then were inoculated by spraying with a conidial suspension (1 × 106 sporongiospores·mL-1). Sterile water served as a control. The test was repeated 3 times. All fruits were cultured at 27°C. Symptoms typical developed on the fruits after 3 days, yellow hyphae covered the entire fruits, but control fruits remained healthy (Fig. 1I, J). The fungus was reisolated from symptomatic tissues and identified as M. inaequisporus by morphology characteristics and molecular characterization. To our knowledge, this is the first report of Fruit rot caused by M. inaequisporus on Chinese plum (Prunus salicina) in China.

ancylostomatid Kalicephalus spp. is the common parasitic intestinal nematode of reptiles. West-Asian blunt-nosed viper is a venomous snake found in extensive areas of Iran. From June to September 2017, two dead viper snakes were referred to a parasitology laboratory and examined for intestinal parasites. Several white elongated roundworms were collected and fixed to identify under light and scanning electron microscopes (SEM) based on morphological and molecular characteristics. For the molecular survey, some parts of the identified worms were extracted and the ITS of nuclear ribosomal DNA (rDNA) was amplified by polymerase chain reaction (PCR). Five roundworms were found in one snake and three worms with similar morphological characteristics in another one. All the collected female hookworms were taxonomically identified as Kalicephalus viperae viperae. The SEM findings showed the head was small and had three dorsal, ventral, and middle circumoral papillae with a spike-like process on the median papilla of K. viperae. Moreover, the buccal capsule was bivalvular and included two lateral valves consisting of several chitonid pieces. The tail of the female worm was slim and long with a blunt end and had a terminal spike at its end. In the molecular survey, the ITS of rDNA amplified at about 850 bp was identified as K. viperae. The ITS gene rDNA phylogeny analysis of the K. viperae sequence showed that the isolated species had high similarity to Ancylostoma species from around the world and is close to Ancylostoma braziliense with 88% discrepancies in the phylogenetic tree. The morphological characteristics and a large part of K. viperea viperea rDNA nucleotide sequence were reported in viper snakes for the first time in the world and in Iran.

Demonstration That Mast Cells, T Cells, and B Cells Bearing the Activating Kit Mutation D816V Occur in Clusters within the Marrow of Patients with Mastocytosis

Lasia spinosa (L.) Thwaites (Araceae) is an ornamental medicinal plant grown in tropical and subtropical countries. The leaf extract of L. spinosa has significant anthelmintic efficacy (Yadav and Temjenmongla 2012). During July 2018 and 2019, L. spinosa plants at the Guangxi Botanical Garden of Medicinal Plants of Guangxi province, China, showed leaves with brown spots or irregular white necrotic lesions that were scattered with some shot holes. Sixty-eight plants from a total of 200 plants showed such symptoms. The overall incidence was about 34%. To isolate the pathogen, 15 infected leaves were excised near the margin of necrotic lesions, surface sterilized with 75% ethanol for 1 min, and treated with 0.5% sodium hypochlorite for 10 min followed with three rinses with sterile water. The leaf sections were then plated on potato dextrose agar (PDA) medium and incubated at 28°C. Further, to obtain pure cultures, hyphal tips from recently germinated spores were excised and transferred onto fresh PDA and incubated at 28°C. Ten fungal isolates with similar morphological characteristics were obtained. Colonies on PDA developed white fluffy aerial mycelia and produced abundant conidia 2 weeks later. Microconidia were abundant, oval, clavate, solitary, and hyaline, with an average size of 7.7 × 3.1 μm (n = 46). Macroconidia were mostly three to five septate, slender, almost straight, and sickle shaped, with an average size of 26.7 × 3.6 μm (n = 30). No chlamydospores were observed. These morphological characteristics were consistent with those of Fusarium fujikuroi (Leslie and Summerell 2006). For further identification, five DNA regions of isolate CYA, corresponding to the internal transcribed spacer (ITS), β-tubulin (TUB2), calmodulin (CL), translation elongation factor 1-alpha (TEF1), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were PCR amplified and sequenced (Jiang et al. 2018; Weir et al. 2012). BLASTN analyses of ITS and CL sequences showed 100% similarity with corresponding sequences of F. fujikuroi accession numbers MK630103.1 and MF984412.1; the TUB2 and TEF1 sequences shared 99.71% identity (MH398245.1 and MH341224.1), and the GAPDH sequence shared 98.65% identity (CP023092.1). Sequences were submitted to GenBank under the accession numbers MN704565 (ITS), MN490089 (TUB2), MN490091 (CL), MN490092 (TEF1), and MN490090 (GAPDH). According to those morphological and molecular characteristics, the pathogen was identified as F. fujikuroi. Further, to verify the pathogenicity, spore suspensions (10⁵ conidia/ml) were prepared from 10-day-old PDA cultures and inoculated onto leaves of nine 2-month-old L. spinosa plants using the wound/drop inoculation method (10 μl per wound). Three more plants were inoculated with sterile water as controls. All plants were grown in a growth chamber with a temperature of 28°C and 90% humidity. After incubating for 7 days, leaf spot symptoms were observed on the inoculated leaves, and no symptoms were observed on the control plants. F. fujikuroi was reisolated from leaf lesions on the inoculated leaves, thus confirming Koch’s postulates. The pathogenic experiment was subsequently repeated under the same conditions. To our knowledge, this is the first report of F. fujikuroi causing leaf spot of L. spinosa. Severe leaf spot disease on L. spinosa has affected its medicinal and ornamental values, which suggests breeders should take this disease into consideration in L. spinosa breeding programs.