Necrotic Patches Research Articles

Characterization of feeding damage by tea mosquito bug, Helopeltis theivora Waterhouse (Hemiptera: Miridae) on Hainan Dayezhong tea cultivar.

The tea mosquito bug, Helopeltis theivora Waterhouse (Hemiptera: Miridae), is a devastating piercing-sucking pest in tropical tea plantations. The Hainan Dayezhong (HNDYZ) is a large-leaf tea cultivar widely cultivated around the Hainan tea region in South China. However, information regarding the feeding damage of H. theivora on the HNDYZ tea plant remains scarce. Here, we first describe the morphology of H. theivora in Hainan tea region. Subsequently, we investigate the feeding biology of H. theivora on HNDYZ tea shoots under laboratory conditions. Additionally, we survey the infestations of H. theivora in a small-leaf Jinxuan tea plantation and three large-leaf HNDYZ tea plantations under varying shaded conditions. The results indicated that the morphological features of eggs, nymphs, and adults of H. theivora in the Hainan tea region were similar to those of the same species reported in other tropical tea regions. Nymphs and adults of H. theivora primarily fed on tender leaves and produced a subcircular spot within 2 to 4 minutes. This feeding spot would gradually turn dark brown within 24 hours. Furthermore, the adjacent scattered spots would connect after 48 hours, resulting in a necrotic patch on the leaves by 72 hours. The peak feeding time for H. theivora occurred at night, specifically from 7:00 PM to 1:00 AM. The most preferred feeding site was at the second leaf position, accounting for 70.94 ± 3.68% of daily feeding spots. During the feeding peak, adults H. theivora produced more feeding spots than nymphs, with females and 5th-instar nymphs creating the largest feeding areas among all life stages. Field investigations showed that damage caused by H. theivora on the large-leaf HNDYZ tea cultivar was significantly greater than that on the small-leaf Jinxuan tea cultivar. More serious infestations of H. theivora were observed in the high-shade HNDYZ tea plantation compared to the medium-shade and no-shade HNDYZ tea plantations. This suggests that the different tea cultivars and shade conditions in tea plantations may influence the population of H. theivora in the field. These findings provide new insights for further research related to the feeding strategy of H. theivora on the HNDYZ tea cultivar.

Comparative Study of the Histopathological Changes of Liver and Lung of Two Lines of laying Chickens Experimentally Infected by Ascaridia galli

This work aimed to determine the effect of A. galli on the histological changes in the liver and lung of two distinct lines of laying chickens and study the resistance of each line. In this study, fifty laying hens 8 weeks of age were used, (25 of which were Lohmann brown-classic and 25 of which were Lohmann lsl-lite). Each line was divided into two groups: the infected group (N=15) these chickens were infected with a single dose of 500 embryonated eggs through oral inoculation, and the second group control group (N=10) was given normal saline. After 8 weeks the chickens were killed to examination the histological changes. The results showed a histopathological lesion of the liver of the Lohmann brown-classic infected group showed fatty degeneration, blood vessels congestion and the fibrin and inflammatory cells in the lumina of the dilated veins these inflammatory cell infiltrations include eosinophil, while Lohmann lsl-lite showed congestion of blood vessels, as well as lymphocyte infiltration of the inflammatory cells. The lung of Lohmann brown-classic showed congestion in inflammatory cell blood vessels and infiltration with alveolar congestion and eosinophils inflammation and necrotic patches, whereas histopathological changes of the lung of Lohmann lsl-lite showed eosinophils proliferation within an alveolar wall, deterioration of their lumina as a result of haemorrhage. While the control group of two-line showed normal liver and lung. Concluded that the A. galli had an effect on the histological structure of the liver and lung, additionally, Lohmann lsl-lite was more affected and less resistant compare to Lohmann brown-classic.

Management of Dustavrana by Chedana Karma and application of Madhu - A Case Study

Dusta Vrana management is most complicated problem encountered by medical practitioner all over the globe, that is non-healing, rapidly growing, disturb the day to day activities of patients, without early and optimal intervention it may ends with amputation of affected limb to save the life, This study reveals that Chedan (debridement) of necrotic patches followed by application of Madhu gives tremendous result in healing of ulcers, As per Sushruta Samhita, Lakshana of Dusta Vrana are Dourganda, Putisrava, Twak Mamsadi Kotha etc., In this case we had Lakshanas like Twak Kotha, Dourganda, Atiputisrava, Kandu etc. In Sushruta Samhita, Chikitsa Sthana 1st chapter Sushruta describes Shasti Upakrama for Vrana Chedana and Madhu Lepana is one among them. Chedana is mainly indicated for Dustavrana, Kotha etc., Honey has properties like Vrana Shodana, Vrana Ropana, Vvrana Sandhaniya. A 45 year male patient with wound over lateral aspect of thigh, knee joint and over lateral aspect of tibia associated with pain, itching fever since 1 week. Patient also had a history of admission in allopathic hospital under diagnosis of cellulitis, where in fasciotomy / fasciectomy was done. As the patient didn’t get relief so he approached our hospitals Shalya Tantra OPD. Patient had no comorbidity like diabetic mellitus, hypertension and patient was treated with Chedana Karma and application of Madhu along with Shamana Aushadi which has been discussed here.

COVID-19 and COVID-19 Vaccine-Related Skin Ulcerations in the Lower Extremities: A Case Report and Literature Review

Several associations have been made between COVID-19 and vasculitis. Recent data also shows the prevalence and association of de novo vasculitis with either COVID-19 infection or COVID-19 post vaccination. In this article, we present the case of new-onset leukocytoclastic vasculitis, secondary to COVID-19 vaccination, that was complicated by severe infected and nonhealing ulcers in the lower extremities. Case A 53-year-old male patient presented to the dermatology clinics with a three-week history of painful necrotic patches coalescent of the lateral malleolus of the right and left ankles. History goes back to when the patient reported developing pruritic papules two weeks after receiving his second shot of the Pfizer BioNTech COVID-19 vaccine (BNT162b2). Punch biopsy was consistent with leukocytoclastic vasculitis. He was prescribed a four-week course of systemic corticosteroids and antibiotics as per cultures. Vascular assessment confirmed normal peripheral arterial and venous system. Two months later, the patient re-presented with fever and worsening of his lower extremity ulcers. He underwent debridement of his wounds. Intra-operative cultures revealed multidrug resistant bacteria. He required an additional debridement session a few days later and a 14-day course of Piperacillin-Tazobactam. The patient was subsequently discharged on corticosteroids and Azathioprine and followed up in the vascular surgery and rheumatology clinics. At four months follow-up, the patient's wounds were almost completely healed. Conclusion This article highlights a case of severe new-onset COVID-19 vaccine-associated leukocytoclastic vasculitis complicated with infected ulcers that required debridement twice in addition to a prolonged course of antibiotics and immunosuppression therapy. To our knowledge, none of the cases reported in the literature were this severe in nature. In this post-pandemic era, it must remain high on the differential list, and healthcare specialists should maintain a high index of suspicion when evaluating sudden new-onset skin lesions that do not have an immediately apparent etiology.

Assessment of the clinical and laboratory risk factors for thrombosis in neonates admitted to neonatal intensive care unit (two Egyptian tertiary centers experience)

In neonates admitted to the neonatal intensive care unit (NICU), arterial and venous thromboembolism is a major cause of morbidity and death which could be attributed to multiple risk factors exposure. This study aimed to evaluate the clinical characteristics, laboratory and radiological assessments, predisposing risk factors, and outcomes of thrombosis in neonates admitted to NICU. This prospective cohort study was conducted at NICU, Minia, and Alexandria University Children’s Hospital. Screening of 886 patients admitted to NICU over one year with different clinical presentations, patients were classified into the thrombotic and non-thrombotic groups based on the presence or absence of thrombosis. Thrombosis was diagnosed based on clinical, laboratory and different radiologic assessments. Genetic testing for factor V Leiden mutations G1691A, prothrombin mutation G20210A, protein C, protein S, and antithrombin III gene mutations were performed for patients with a family history of thrombosis. Out of a total of 886 neonatal admissions, 36 patients were diagnosed with evident thrombosis (40 per 1000 NICU admissions). The sites of venous thrombosis detection were Portal vein thrombosis in 11 patients (30.6%), superior vena cava thrombosis in 7 patients (19.4%), deep venous thrombosis in 5 patients (13.9%), central venous thrombosis in 5 patients (13.9%), intra-cardiac thrombosis in 3 patients (8.3%) and necrotic skin patches in one patient (2.8%). Only 69% of enrolled thrombosis patients showed genetic mutations the most common of which was factor V Leiden mutation (52.3%). Sepsis, central venous line (CVL) insertion, C reactive protein (CRP), and duration of NICU admission were significantly more common in the thrombotic group (p < 0.001) and were associated with a higher risk of thrombosis (ORs: 1.02, 7.7, and 1.11, respectively) (p < 0.001). Higher mortality occurred in thrombosis neonates compared with a non-thrombotic group (52.8% versus 17.4%) (p < 0.001). NICU-admitted neonates are exposed to multiple overlapped risk factors, the detection of which is important for preventing potential thrombosis and improving the patient’s outcomes. The complexity of sepsis pathogenesis and management could potentiate multiple acquired risk factors. inherited thrombophilia detection is required for prevention of further morbidities.

314. TRANSORAL ANTEGRADE T-TUBE INSERTION FOR MANAGEMENT OF OESOPHAGEAL LEAKS FOLLOWING MINIMAL INVASIVE OESOPHAGECTOMY

Abstract Introduction Oesophageal leaks following minimal invasive oesophagectomy (MIE) present significant challenges to upper GI surgeons, with reported rates ranging from 11% to 21%. Management strategies include: conservative, endoscopic, and surgical interventions. These vary based on patient clinical status and institution. Here, we propose a hybrid approach involving thoracoscopic washout coupled with antegrade T-tube insertion via gastroscopy for managing these complex leaks. Methods We describe the case of a 66-year-old male who underwent MIE via the McKeown approach for Siewert 1 Gastro-oesophageal adenocarcinoma. His neoadjuvant chemotherapy had been interrupted by a complex ischiorectal abscess and thus a decision was made to proceed for surgery. The surgery proceeded with a cervical anastomosis using a linear stapled technique and closure with 3.0 PDS. Postoperatively, the patient deteriorated on day 2, necessitating intubation due to decreased oxygen saturation. The chest tube output looked slightly bile stained. A subsequent CT scan revealed mediastinal fluid accumulation, prompting urgent intervention. Results Diagnostic thoracoscopy revealed contamination of the right chest and posterior mediastinum, necessitating extensive washout. A 2cm necrotic patch and perforation were identified on the gastric side of the oesophago-gastric anastomosis. Intraoperative gastroscopy localized the leak with a positive air bubble test. A Maryland forceps was passed through the defect to confirm the defect. Subsequently, a guidewire was passed antegrade into the cavity using the gastroscope. Decision was made to purse-string the defect (around the guidewire) using 3.0 PDS and the T-tube was exchanged transorally via the guidewire into the cavity and exteriorized through a 5mm port to the skin. Existing large-bore chest and Blake drains were retained, and a feeding jejunostomy was established. Conclusion This hybrid management approach, combining endoscopy and thoracoscopic washout with T-tube insertion, offers an alternative for addressing complex anastomotic leaks following MIE. By avoiding resection and diversion, this technique potentially reduces morbidity. Follow-up scans demonstrated resolution of the leak allowing the patient to be discharged. https://1drv.ms/v/s!Aghd8kXfo8oQlX6QyliVLgRDmIu-?e=EGN5YD

Clinical Presentation and Treatment Outcomes of Necrotizing Fasciitis: A Prospective Comparative Study of Children and Adults in Sokoto.

Necrotizing fasciitis (NF) is a severe soft tissue infection typified by swiftly spreading necrosis of the fascia and subcutaneous fat with successive necrosis of the skin which affects all age groups. To compare the clinical presentation and treatment outcome of NF between children and adults. A prospective descriptive study of all patients presenting with NF to the (Usmanu Danfodiyo Univrersity Teaching Hospital, Sokoto), from September 2018 to August 2019. Participants were stratified into adults and children for comparison during data analysis. A total of 32 patients were analyzed, 24 (75.0%) were adults. The age range was 2 weeks to 75 years with a median age of 35 years. Necrotic skin patches and skin erythema were the most common cutaneous findings. The lower limb was the most commonly affected region in adults (58.3%), whereas the head and neck and abdominal wall were equally commonly involved in children (37.5%). Children had more multiple isolates than adults, 87.5% compared to 62.5%, respectively, and it was significant (P value 0.04). Escherichia coli and Klebsiella species were both significantly commoner in isolates of children than adults (P value 0.01 and 0.04, respectively). The method of wound coverage was significantly different (P value 0.003) as fewer children required skin grafting. The mortality rate was 6.25%, the two deaths occurred in adults. NF is commoner in adults in our region. The predilection sites of the disease in the two groups differ and healing by secondary intention was more possible in children.

Roles of the Arabidopsis KEULE Gene in Postembryonic Development.

Cytokinesis in plant cells begins with the fusion of vesicles that transport cell wall materials to the center of the cell division plane, where the cell plate forms and expands radially until it fuses with the parental cell wall. Vesicle fusion is facilitated by trans-SNARE complexes, with assistance from Sec1/Munc18 (SM) proteins. The SNARE protein KNOLLE and the SM protein KEULE are required for membrane fusion at the cell plate. Due to the crucial function of KEULE, all Arabidopsis (Arabidopsis thaliana) keule mutants identified to date are seedling lethal. Here, we identified the Arabidopsis serrata4-1 (sea4-1) and sea4-2 mutants, which carry recessive, hypomorphic alleles of KEULE. Homozygous sea4-1 and sea4-2 plants are viable and fertile but have smaller rosettes and fewer leaves at bolting than the wild type. Their leaves are serrated, small, and wavy, with a complex venation pattern. The mutant leaves also develop necrotic patches and undergo premature senescence. RNA-seq revealed transcriptome changes likely leading to reduced cell wall integrity and an increase in the unfolded protein response. These findings shed light on the roles of KEULE in postembryonic development, particularly in the patterning of rosette leaves and leaf margins.

Unveiling Dominant Fungal Pathogens Associated with Root Rot of Hybrid Bermudagrass Based on Culture Dependent and Independent Methods.

Hybrid bermudagrass (Cynodon dactylon × C. transvaalensis) is widely used as turf in southern and transition zones of China. From June to September in 2022, an unknown disease was consistently observed on hybrid bermudagrass in different regions of Nanjing, China, that exhibited distinct symptoms of leaf necrosis, severe root rot, and circular or irregular necrotic patches with 20- to 300-cm diameters. In this study, culture-independent and -dependent methods were used to elucidate the dominant fungal pathogens associated with the disease. Basidiomycota and Marasmiellus were shown to be the dominant phyla (51.96 to 70.60%) and genera (50.09 to 69.84%) in the symptomatic samples. A total of 128 fungal strains were isolated from symptomatic root tissues, and 40 strains representing the largest proportion (31.25%) were identified as Marasmiellus mesosporus based on the morphological characteristics, phylogenetic analysis of the ITS and LSU rDNA region, and pathogenicity testing. Temperature sensitivity tests revealed that M. mesosporus grew well at high temperatures (growth rate of 13.74 mm/day at 36°C). To our knowledge, this is the first report of M. mesosporus causing root rot disease on hybrid bermudagrass during hot summer months. The study will have important implications for the management of the disease.

A necrotic patch on the elbow of a child.

A necrotic patch on the elbow of a child.

Genetic characterisation of canine adenovirus type 1 in an 18‐month‐old mixed breed male dog and its effect on clinico‐pathology, hemato‐biochemistry and serum oxidant–antioxidant status

AbstractAn 18‐month‐old, mixed breed male dog was presented with hypothermia, recumbency, hematemesis, melena, and moribund condition, and screened by rapid antigen detection kit followed by polymerase chain reaction confirmation for canine adenovirus type 1 infection. The phylogenetic analysis of the sequence revealed clustering with Indian, Spanish, Italian, Portuguese, Chinese, and Australian sequences with amino acid homology of 98.81%–100%. There was a mutation (N127D) in the E3 protein encoding sequence, which was hitherto reported only in one Indian (GQ340423) sequence. Haemogram revealed anaemia, neutropenic leukopenia, and thrombocytopenia. Serum biochemistry revealed marked hyperglycaemia, hypoproteinemia, hypoalbuminemia, and an increase in the activities of hepatic enzymes. Serum oxidant antioxidant parameters revealed a marked increase in the level of oxidant and a decreased level of antioxidants. Postmortem examination revealed multifocal necrotic patches in the liver, marked haemorrhage in the stomach and intestines, emphysematous lungs, splenomegaly, and congested kidney. Histopathology revealed necrohaemorrhagic hepatitis and intranuclear inclusion bodies in the hepatocytes and glomerular degeneration.

Characterization of Athelia rolfsii associated with southern blight of pointed gourd (Trichosanthes dioica) in India

Pointed gourd (Trichosanthes dioica Roxb) is a perennial cucurbitaceous vegetable popular in the eastern and northern parts of India. In July 2018, an occurrence of southern blight disease was observed in a pointed gourd crop grown in Odisha state located in the eastern part of India. The typical symptoms included angular necrotic patches and disintegration of leaf tissues with the vein and veinlet intact, exhibiting a diagnostic netted pattern of veins. Silky white, fan-shaped mycelial growth with sclerotial bodies was observed on the lower surface of infected leaves. Eventually, the infected leaves dried up, leaving several bare patches in the field. Associated fungi were isolated, which resembled the reference strain Athelia rolfsii based on colony characteristics. Phylogenetic analysis of the internal transcribed spacer of rRNA gene region confirmed that the isolates were A. rolfsii. Pathogenicity was successfully proved on detached leaves as well as on potted pointed gourd plants. On the basis of symptoms associated with the disease, fungal morphology, molecular identification, and pathogenicity tests on host plants, the causal fungus was identified as A. rolfsii. This is the first report of southern blight disease caused by A. rolfsii on pointed gourd in India.

A CASE STUDY ON THE PRE-GANGRENOUS STAGE FOLLOWED BY LEECH THERAPY AND AYURVEDA INTERVENTION OF KAISHORE GUGGULU

Around the world, a lower leg is amputated every 30 seconds due to gangrene. The mortality rate from diabetic foot gangrene is almost precisely the same as the mortality rate from cancer. When gangrene becomes severe, the only alternative treatments are surgical debridement of the injured tissue (which cannot be reversed) or amputation of the affected limb. The current case study provides an alternative to conventional medical care for chronic wounds &amp; the gangrenous stage that is not healing correctly: Ayurvedic medicine (blood purifier and oxygen-sufficiency enhancer). A 60-year-old woman who had developed a chronic wound which was eventually converting into dry gangrene alongside intense pain was in danger as she was advised to have her left foot amputated below the knee underwent this trial. The patient's gangrenous great toe was extremely painful (80 mm on a 100 mm visual analogue scale) and had necrosed modifications that smelled bad. Aragvadhadi kashaya &amp; Shatdhaut Ghrita were used as wound dressing since it is particularly effective at removing necrotizing areas. Leech Therapy or Hirudotherapy, along with Kishore Guggulu, were used for pain management together to promote healing, oxygen sufficiency &amp; rejuvenation of the skin. Within 25 days, the pain score on a 100 mm visual analogue scale dropped to 0-10 mm, and no more painkillers were needed. Necrotic patches vanished, and the wound healed entirely over almost eight weeks (2 months)

First report of anthracnose caused by Colletotrichum karstii on Dalbergia odorifera in China.

Dalbergia odorifera (Family: Fabaceae) is a national second-grade protected tree in China with high medicinal and economic value (Zhao et al., 2020). In July, 2022, a leaves spot disease on D. odorifera with typical anthracnose symptoms was observed in plantations in Haikou (110.319153°E, 19.072900°N), Dongfang (108.630297°E, 19.103838°N) and Qiongzhong (109.704460°E, 19.088440°N), Hainan Province, China. Disease incidence was 7.5% (n = 50 plants). Early symptoms of infected leaves were small and round dark brown spots, which developed into larger irregular necrotic lesions and leaves withered. Leaf tissues (5×5 mm) at the disease-health junction of spots from 19 leaves were sterilized with 2.5% sodium hypochlorite for 1 min, and rinsed with sterile distilled water three times. These sterilized tissues were placed on potato dextrose agar (PDA) and incubated at 28 ℃ for 5 d. 7 strains of fungi with similar morphology were isolated, and 3 single-hyphal isolates (HHL01, HHL02 and HHL03) from each location were selected for further study. Colonies on PDA were fluffy orange-yellow mycelium. Conidia were aseptate, cylindrical, smooth-walled, straight, hyaline with both ends bluntly rounded, 11.82 to 15.77 × 3.87 to 6.71 μm (n = 100; average = 13.75 × 5.52 μm). Appressoria formed on slides, measured 5.54 to 10.64 × 4.19 to 7.41 μm (n = 30; average = 8.06 × 5.97 μm) were brown to black, elliptical to irregular. For molecular biological identification, the genomic DNA of three isolates was extracted by fungal genomic DNA extraction kit (Tiangen Biotech (Beijing) Co., Ltd.). The partial sequences of internal transcribed spacer region (ITS; ITS1/ITS4), glyceraldehyde-3-phosphate dehydrogenase (GAPDH; GDF1/GDR1), actin (ACT; ACT512F/ACT783R), β-tubulin (TUB2; TI/Bt2b) and calmodulin (CAL; CL1C/CL2C) were amplified and sequenced by Sangon Biotech (Shanghai) Co., Ltd (Carbone and Kohn, 1999; Weir et al., 2012). The sequences were deposited as GenBank Accession Nos. OR018110-OR018112 (ITS); OR050529-OR050537 (GAPDH, ACT and CAL) and OR192168-OR192170 (TUB). BLASTn results showed these sequences were more than 99% identity with the strain of C. karstii CORCK1 (GenBank Accession Nos. HM585406, HM585387, HM581991, HM585424 and HM582010, respectively). Multi-locus phylogenetic tree of Colletotrichum spp. showed that those three isolates were sister to C. karstii based on the maximum likelihood and bayesian inference methods. To verify pathogenicity, 2 mL spore suspension (1 × 106conidia/ml) of the isolates was sprayed on each leaves of 1-year-old D. odorifera plants, and sterile distilled water was similarly sprayed on other leaves as a negative control. The plants were incubated in a greenhouse under 90% ± 5% RH at 28 °C. Light brown small round necrotic patches developed 3 days after inoculation, while the control was asymptomatic. Photographs were taken on the fifth day after inoculation. The fungi were re-isolated from the diseased leaves and identified by morphological characterization and molecular identification, fulfilling Koch's postulates. C. karstii has been reported causing leaf rot of Carissa grandiflora in Spain (Garcia-Lopez et al., 2021), and anthracnose caused by C.tropicale was reported on D. odorifera (Yi et al., 2023). To our knowledge, this is the first report of Dalbergia odorifera leaf spot disease caused by Colletotrichum karstii. This finding provides an important basis for further research on the control of the disease.

First Report of Alternaria alternata Causing White Leaf Spot on Allium tuberosum in China.

Puding County is the major Allium tuberosum growing area in Guizhou Province of China. In 2019, white leaf spots were observed on Allium tuberosum in Puding County (26.31°N, 105.64°E). The white spots, ranging from elliptic to irregular in shape, first appeared on leaf tips. With disease aggravation, spots gradually coalesced, forming necrotic patches with yellow margins causing leaf necrosis; sometimes there was gray mold on dead leaves. The incidence of the diseased leaf rate was estimated to be 27-48%. To identify the pathogenic agent, 150 leaf tissues (5 mm × 5 mm) were obtained from disease-healthy junctions of 50 diseased leaves. Leaf tissues were disinfected in 75% ethanol for 30 s, soaked in 0.5% sodium hypochlorite for 5 min, and flushed three times with sterile water, before being placed on potato dextrose agar (PDA) in the dark at 25 °C. When colonies appeared, the mycelial tips were picked and placed on new PDA. Purified fungus was obtained after repeating this last step several times. The colonies were grayish-green with white round margins. Conidiophores (2.7-4.5 μm × 27-81 μm) were brown, straight, or flexuous with branches and septa. Conidia (8-34 µm × 5-16 µm) were brown, with 0-5 transverse septa and 0-4 longitudinal septa. The 18S nuclear ribosomal DNA (nrDNA; SSU), 28S nrDNA (LSU), RNA polymerase II second largest subunit (RPB2), internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and translation elongation factor 1-alpha (TEF-α) (Woudenberg et al. 2013) were amplified and sequenced. The sequences were deposited in GenBank (ITS: OP703616, LSU: OP860684, SSU: OP860685, GAPDH: OP902372, RPB2: OP902373, TEF1-α: OP902374). According to BLAST analysis, the ITS, LSU, GAPDH, RPB2, SSU, and TEF1-α of the straishowed 100% (689 of 731 base pairs; bp), 100% (916 of 938 bp), 100% (579 of 600 bp), 100% (946 of 985 bp), 100% (1093 of 1134 bp), and 100% (240 of 240 bp) sequence identity to those of Alternaria alternata (ITS: LC440581.1, LSU: KX609781.1, GAPDH: MT109295.1, RPB2: MK605900.1, SSU: ON055699.1 and TEF1-α: OM220081.1). A phylogenetic tree was constructed using PAUP4 and the maximum parsimony method with 1000 replicas of bootstrapping for all datasets. According to morphological characteristics and phylogenetic analysis, FJ-1 was identified as Alternaria alternata (Simmons 2007, Woudenberg et al. 2015). The strain was preserved in the Agricultural Culture Collection of China (preservation number: ACC39969). To determine the pathogenicity of Alternaria alternata against Allium tuberosum, wounded healthy leaves were inoculated with a conidial suspension (106 conidial/mL) and round mycelial plugs (4mm). Sterile agar PDA plugs with no mycelium or sterile water were inoculated as negative controls. Three days later, white spots appeared on the wounded leaves inoculated with mycelial plugs or conidial suspension. However, the symptoms caused by conidial suspensions were weaker than those caused by mycelial plugs. No symptoms were observed in the control group. The experimental symptoms were consistent with the phenomena observed in the field. The same fungus was reisolated from necrotic lesions and identified as Alternaria alternata using the method described above. To our knowledge, this is the first report of Alternaria alternata causing white leaf spots on Allium tuberosum in China, a disease seriously affected the yield and quality of Allium tuberosum and caused economic losses to farmers. Reference: Simmons EG (2007) Alternaria: an identification manual. CBS Fungal Biodiversity Centre, Utrecht, the Netherlands. Woudenberg JHC, Groenewald JZ, Binder M, Crous PW ( 2013) Alternaria redefined. Stud Mycol, 75: 171-212. https://doi.org/10.3114/sim0015. Woudenberg JHC, Seidl MF, Groenewald JZ, Vries M de, Stielow JB, Thomma BPHJ, Crous PW (2015) Alternaria section Alternaria: Species, formae speciales or pathotypes? Stud Mycol, 82:1-21. https://doi.org/10.1016/j.simyco.2015.07.001.

Fig Rust Caused by Cerotelium fi ci: Past, Present and Future

Fig rust is one of the serious fungal diseases caused by Cerotelium fi ci which causes great loss to the overall growth and yield of the plant. This disease is generally characterized by pale reddish-brown to reddish-brown angular leaf spots, typically found on the adaxial surfaces of the leaves, whereas the corresponding abaxial surface of the same leaves was covered with a lot of orange rust pustules. With the progression of this rust disease, angular spots fused to form necrotic patches whereas, leaf edges curled upward. Premature abscission of leaves takes place, leaving bare branches and unripe fruits. This disease showed its global distribution in tropical and subtropical areas, however, the highest distribution was found in the African continent (18 countries) followed by Asia (14 countries), North &amp; South America (10 countries each), Europe (8 countries) and 7 countries in the Oceanian continent. Apart from the rust pathogen Cerotelium fi ci, the Phakospora nishidana was also reported to cause fi g rust which has created confusion on the identity of C. fi ci as a fi g rust pathogen. This suggests a need to re-examine all specimens of fi g rust at the global scale, based on morphological and modern DNAbased methods so that the taxonomic ambiguities on fi g rust pathogens can be resolved and suitable control methods can be implemented to avoid serious crop and yield loss.

A PROSPECTIVE STUDY OF EPIDEMIOLOGICAL FACTORS AND OUTCOME IN PATIENTS OF DIABETIC FOOT WOUND ATTENDING J.A. GROUP OF HOSPITALS: A TERTIARY CARE CENTRE

Objectives: To study about epidemiological factors related to diabetic foot wound such as age, sex, duration of diabetes, relation to ongoing retinopathy, and nephropathy. Methods: After obtaining approval from ethical committee, the present study is to be conducted on 100 patients of diabetic foot in the Department of Surgery, JA Group of Hospitals and GR Medical College, Gwalior (MP) during January 2020 to June 2021 after getting written informed consent from the patients. Results: In our study, out of 100 cases, most of the diabetic foot wound cases observed were in the 50–59 years age group with a mean age of 52.23 and a standard deviation of ±14.92. Out of 100 cases, 82 (61.%) were male and 29 (38.66%) were female. Therefore, male: female ratio was 4.5:1. In our study of 100 patients, 70 (70%) presented with ulcer with or without necrotic patch, gangrene and 30 (30%) with cellulitis with or without abscess. Among all patients of diabetic foot, wound 62 (62%) were associated with retinopathy and 47 (47%) were associated with diabetic nephropathy. Conclusion: Diabetic foot wound is more common in men due to their increased susceptibility to trauma and occupation. Diabetic patients at risk for foot complication must be educated about risk factors and the importance of foot care, including the need for self-inspection and surveillance, monitoring, daily foot hygiene, use of proper footwear, good diabetes control, and prompt recognition and early standard treatment of newly discovered lesions.

Culm blight on Phyllostachys aureosulcata 'Spectabilis' caused by Apiospora locuta-pollinis in China.

Phyllostachys aureosulcata McClure 'Spectabilis' C.D. Chu. et C.S. Chao is predominantly native to subtropical to warm temperate areas and is widely cultivated for landscaping in China (Neményi et al. 2015). In November 2020 (10 - 16 ℃), culm blight symptoms were observed on P. aureosulcata 'Spectabilis' in Wangjiang Tower Park (all kinds of plant areas are about 9.8 ha), Chengdu City (104°09'30.42″ E, 30°63'18.89″ N). Fifty plants were surveyed, and disease incidence was recorded as approximately 30%. Initially, chlorotic necrotic patches appeared on the culms, and gradually the patches became white, expanded to both ends, and encircled the whole culm with black edge and conidiomata, which eventually led to wilt and death. Five samples from different bamboos were collected and one of them were used for morphological observation. Five single conidia isolates were carried out on potato dextrose agar (PDA) at 25±1℃ (Chomnunti et al. 2014). Colonies were initially white and then yellowish in the center with abundant aerial mycelia. On the culm, conidiomata were dry, black, and filamentous. Conidiophores were reduced to conidiogenous cells. Conidiogenous cells were smooth, hyaline, ampulliform to doliiform. Conidia were ellipsoid to globose, dark brown, smooth and aseptate, measuring 5.2 to 9.4 × 4.4 to 7.3 μm, (=8.2 × 6.5μm, n=50). On the PDA medium, conidia were globose to subglobose, olive green to pale brown, and smooth, larger than those from the host in size, measuring 9.0 to 18 × 7.5 to 9.5 μm ( =36.6 × 18.8 μm, n=50). These asexual structures were extremely similar to Apiospora locuta-pollinis (F. Liu & L. Cai) X.G. Tian & Tibpromma (Zhao et al. 2018). DNA was extracted from the representative strain (SICAUCC 22-0036), and the internal transcribed spacer (ITS), translation elongation factor 1-alpha (tef1-α), beta-tubulin (tub2), 28S large subunit rDNA (LSU) were amplified and sequenced with primers ITS1/ITS4 (White et al. 1990), EF1-728F (Carbone & Kohn 1999)/EF2 (O'Donnell et al. 1998), T1 (O'Donnell & Cigelnik 1997)/Bt2b (Glass & Donaldson 1995) and LR0R/LR5 (Rehner & Samuels 1994). The newly generated sequences were deposited in GenBank with accession nos. ON228609 (ITS), ON324018 (tef1-α), ON237657 (tub2), and ON228665 (LSU). Nucleotide blast showed 98.97%, 100% and 99.46% identities with A. locuta-pollinis (LC11683, ex-holotype) (accession nos. MF939595, MF939622, MF939616), and LSU data missing. Phylogenetic analyses using maximum likelihood showed a 92% bootstrap support value in a clade with A. locuta-pollinis (Fig 2). Eight healthy plants (2-year-old) were used for the pathogenicity test. Culms of four healthy bamboos were wounded via sterile double-edged blade and sprayed with conidial suspension (105 conidia/ml) prepared from 4-week-old cultures that were incubated on PDA at 25℃. The other four bamboos were sprayed with sterile distilled water as controls. Inoculated plants were placed in a growth chamber (25℃, 90% relative humidity, 12-h photoperiod). About 60 days later, necrotic patches similar to those observed in the field were found on the inoculated culms, and no symptoms were observed on the controls. The pathogen was reisolated from the diseased culms with identical morphology as previously described. To our knowledge, this is the first report of culm blight on P. aureosulcata 'Spectabilis' caused by A. locuta-pollinis. The risk of this pathogen needs further evaluation, and effective control measures should be taken.

Genetic Diversity and Pathogenicity of Fusarium fujikuroi Species Complex (FFSC) Causing Sugarcane Pokkah Boeng Disease (PBD) in China.

Pokkah boeng disease (PBD), a sugarcane foliar disease, is caused by various Fusarium spp. within the Fusarium fujikuroi species complex (FFSC). In the current study, we investigated the diversity of Fusarium spp. associated with PBD in China. In total, 320 leaf samples displaying PBD symptoms were collected over 10 consecutive years (2012 to 2021), during winter and summer, from six various sugarcane-growing regions (Guangxi, Yunnan, Guangdong, Zhejiang, Hainan, and Fujian) in China. Phylogenetic analysis of Fusarium spp. was reconstructed using translation elongation factor 1-α, and DNA-directed RNA polymerase II largest subunit and second-largest subunit multigene sequences. Evolutionary studies of these regions categorized the isolates into four FFSC species (F. sacchari, F. proliferatum, F. verticillioides, and F. andiyazi). The identified isolates, which developed irregular necrotic patches and rotting symptoms on the sugarcane plant after approximately 30 days were tested for their pathogenicity. Symptoms that appeared during pathogenicity testing were consistent with those observed under field conditions. Each strain of the pathogenic Fusarium spp. belonged to different vegetative compatibility groups (VCGs), and there was no affinity between VCGs. Our results contribute to understanding FFSC and accurately identifying Fusarium spp. associated with the sugarcane crop.

First Report of Fusarium commune Associated with a Root Rot of Grapevine in China.

During last decade, species belonging to Fusarium, Rosellinia, Armillaria and Dactylonectria were confirmed as phytopathogens causing grapevine root diseases (Highet and Nair 1995; Teixeira et al. 1995; Calamit et al. 2021; Ye et al. 2021). From 2020 to 2021, grapevine decline was observed in several vineyards in Beijing region, China. Leaves turned yellow with brown necrotic patches and roots were poorly developed, which was suggesting that a root disease was affecting the vines. The disease incidence was up to 10-15% of the vineyard for sample collection. Symptomatic root samples (cv. 'Red Globe') were collected and tissue fragments were excised at the margin of the symptomatic tissue in order to isolate the potential pathogen. The surface was sterilized using 1.5% sodium hypochlorite for 3 min, followed by 70% ethanol for 30 sec, and rinsed three times with sterile distilled water (Ye et al. 2020). Tissues were dried and placed onto potato dextrose agar (PDA) plates, followed by incubation at 25°C under dark conditions for 3 d. Hyphal tips of fungi growing from the samples were transferred onto new PDA plates and incubated until they produced conidia. Next, single spores were transferred onto new PDA plates and incubated at 25°C for 7 d. Eight isolates numbered with JZB3110172 to JZB3110179 were obtained and their culture characters were identical, and the re-isolation percentage was 100%. Colonies were white to orange, with abundant fluffy aerial mycelium. Macroconidia were fusiform with a slightly curved apical cell and a foot-shaped basal cell, and measured 16.2-43.2 μm × 2.7-4.9 μm (n=50); microconidia were cylindrical, straight to slightly curved, 5.1-13 × 2.1-3.9 μm (n=50). Morphological characters of the isolates resembled to Fusarium commune (Skovgaard et al. 2003). For phylogenetic analysis, genomic DNA of the eight isolates was extracted with a DNA extraction kit (DNeasy plant Mini Kit). PCR amplifications of two phylogenetic markers (EF-1α and RPB2) were performed using the primers EF-1/EF-2 (Geiser et al. 2004) and RPB2-5F2/RPB2-7cR (Liu et al. 1995), respectively. The sequences were deposited in GenBank ON457645 to ON457660. Comparison of base pairs on Maximum likelihood (ML) phylogenetic analysis was conducted using the RAxML-HPC2 tool on XSEDE on the CIPRES Science Gateway platform (http://www.phylo.org/). The sequences of EF-1α and RPB2 of the eight isolates showed 99 to 100% similarity to the reference isolates of F. commune. In the phylogenetic tree, the isolates from this study clustered with the representative strains of F. commune (NRRL 52764, NRRL 28387 and NRRL 52744). Based on morphological characters and the phylogenetic results, all of isolates were identified as F. commune. Koch's postulates were conducted on healthy, 3-month-old grapevine 'Marselan'. Plant roots were trimmed with sterile scissors and then soaked in a spore suspension (1.0 × 106 spores mL-1) or sterile water (as the control) for 30 min. The inoculated grapevines were transplanted into pots and kept in the greenhouse at 25°C. After 14 days, all the inoculated plants developed necrosis and turned yellow. No symptoms were observed on the control. Koch's postulates were fulfilled by re-isolating the fungus from necrotic root tissues. The isolates obtained from the artificially infected tissue were identified again as F. commune based on morphological and molecular analyses. Overall, this is the first report of F. commune associated with a grapevine root rot globally, which lays a foundation for further study and developing disease control methods.