Epidemiology and management of Fusarium wilt of Eucalyptus camaldulensis through systemic acquired resistance.

Eucalyptus camaldulensis is a multifunctional tree and is globally used for the reclamation of problematic lands. Eucalyptus camaldulensis is prone to attack by a number of pathogens, but the most important threat is the Fusarium wilt (Fusarium oxysporum). Keeping in view the importance of E. camaldulensis and to manage this disease, five plant activators, i.e., salicylic acid (C7H6O3), benzoic acid (C7H6O2), citric acid (C6H8O7), dipotassium phosphate (K2HPO4), monopotassium phosphate (KH2PO4) and nutritional mixture namely Compound (NPK) and nutriotop (Fe, Zn, Cu, B, Mn) were evaluated in the Fusarium infested field under RCBD in the Research Area, Department of Forestry and Range Management, University of Agriculture, Faisalabad (UAF). Among plant activators, salicylic acid and a combination of compound + nutriotop exhibited the lowest disease incidence and enhanced fresh and dry weight of leaves compared to other treatments and control. Results of the environmental study indicated maximum disease incidence between 35-40 °C (max. T), 6-25 °C (mini. T), 70-80% relative humidity and 1.5-2.5 km/h wind speed while pan evaporation expressed weak correlation with disease development. It was concluded that Fusarium wilt of Eucalyptus camaldulensis could be managed through activation of the basal defense system of the host plant with provision of salicylic acid and balanced nutrition by considering environmental factors. Recent exploration is expected to be helpful for future research efforts on epidemiology and ecologically sound intervention of Fusarium wilt of Eucalyptus camaldulensis.

Hydrochemical, hydrogeological, and cancer health risk analysis of groundwater due to anthropogenic activities, Hasilpur, Pakistan.

Drinking water quality deteriorates rapidly due to anthropogenic activities and rapid population growth. These activities, in developing countries, will lead to water scarcity. In Pakistan, 70% of the population has no access to safe water, and people use canal water to drink. This study performed hydrochemical, hydrogeological, and cancer risk analyses on Tahsil Hasilpur, Bahawalpur, Pakistan. Thirteen tube wells were selected for groundwater and borehole log study. Twenty-two drinking water quality parameters were analyzed using standard methods and quality checks. The borehole data (2D and 3D) shows the abundance of sand (fine and coarse) with some uniformities, which changes the groundwater quality. The results of water quality parameters show that the concentration of TDS (2064-11,159 mg/L), Cl-1 (213-4917 mg/L), As+3 (0.048-0.158 mg/L), Pb+2 (1.294-1.673 mg/L), and Cd+2 (0.008-0.053 mg/L) were beyond guideline values. The statistical analysis showed that the parameters have a moderate to strong correlation (Pearson correlation), which may be due to the same origin (ANOVA). The principal component analysis and cluster analysis confirm the multiple sources of pollutants in the groundwater of the study area. The Piper, Durov, Stiff, and Scholler diagrams confirm that the groundwater system has an abundance of Ca+2 and Mg+2 with Cl-1. The Gibbs diagram showed that the groundwater is not saturated and tends to dissolve more minerals. The hazard quotient values are above 1.0, which indicates noncancer risk severity. The HQ trend was As+3 > Pb+2 > Cd+2 > Ni+2 > Cu+2 > Cr+2 > Zn+2 > Fe+2. The cancer risk values showed that 3-5 people/100 population were exposed to cancer risk. The trend of CR was As+3 > Cd+2 > Cr+2 > Pb+2 > Ni+2. The GIP mapping of pollutants showed that the concentration of pollutants near the canals was high compared to the locations away from the canal. The overall groundwater quality is alarming and needs immediate government attention.

An eco-friendly hydroentangled cotton non-woven membrane with alginate hydrogel for water filtration.

Alginate hydrogel is highly efficient for water filtration due to its anti-fouling nature and formation of strong hydration membranes. However, poor mechanical properties of alginate hydrogel membrane limit its installation in water treatment. There is a need to enhance mechanical properties of alginate hydrogel membranes using eco-friendly, cost-effective materials and technologies. In this work, hydroentangled non-woven from cotton waste (comber noil) fibers was prepared. This non-woven was immersed in solution of sodium alginate (0.5 %, 1 %, 1.5 %) followed by dipping in calcium chloride solution which resulted in gel formation on and into cotton fibers. The successful formation of gel on non-woven fabric was confirmed through FTIR (Fourier transform infrared spectroscopy) and properties of this composite membrane were analyzed by SEM (Scanning electron microscopy), XRD (X-ray diffraction), DSC (Differential scanning calorimeter), water contact, water flux, oil-water filtration, air permeability, tensile strength, and porosity tests. The results showed that porosity of prepared hydrogel membranes decreased with increasing alginate concentration from 0.5 % to 1.5 % which resulted in decreased water permeation flux from 2655 h-1/m2 to 475 h-1/m2. The prepared membrane has separation efficiencies for the oil-water mixture in the range of 97.5 % to 99.5 %. Moreover, the developed samples also showed significant antibacterial activity as well as improved mechanical properties. The strength of the prepared membrane is in the range of 40 N to 80 N. The developed sodium alginate hydrogel-based non-woven membrane could have potential applications for commercial water filtration systems.

Comprehensive genomic analysis of Bacillus paralicheniformis strain BP9, pan-genomic and genetic basis of biocontrol mechanism.

Many Bacillus species are essential antibacterial agents, but their antibiosis potential still needs to be elucidated to its full extent. Here, we isolated a soil bacterium, BP9, which has significant antibiosis activity against fungal and bacterial pathogens. BP9 improved the growth of wheat seedlings via active colonization and demonstrated effective biofilm and swarming activity. BP9 sequenced genome contains 4282 genes with a mean G-C content of 45.94% of the whole genome. A single copy concatenated 802 core genes of 28 genomes, and their calculated average nucleotide identity (ANI) discriminated the strain BP9 from Bacillus licheniformis and classified it as Bacillus paralicheniformis. Furthermore, a comparative pan-genome analysis of 40 B. paralicheniformis strains suggested that the genetic repertoire of BP9 belongs to open-type genome species. A comparative analysis of a pan-genome dataset using the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Cluster of Orthologous Gene groups (COG) revealed the diversity of secondary metabolic pathways, where BP9 distinguishes itself by exhibiting a greater prevalence of loci associated with the metabolism and transportation of organic and inorganic substances, carbohydrate and amino acid for effective inhabitation in diverse environments. The primary secondary metabolites and their genes involved in synthesizing bacillibactin, fencing, bacitracin, and lantibiotics were identified as acquired through a recent Horizontal gene transfer (HGT) event, which contributes to a significant part of the strain`s antimicrobial potential. Finally, we report some genes essential for plant-host interaction identified in BP9, which reduce spore germination and virulence of multiple fungal and bacterial species. The effective colonization, diverse predicted metabolic pathways and secondary metabolites (antibiotics) suggest testing the suitability of strain BP9 as a potential bio-preparation in agricultural fields.

Nanobiotechnology in crop stress management: an overview of novel applications.

Agricultural crops are subject to a variety of biotic and abiotic stresses that adversely affect growth and reduce the yield of crop plantss. Traditional crop stress management approaches are not capable of fulfilling the food demand of the human population which is projected to reach 10 billion by 2050. Nanobiotechnology is the application of nanotechnology in biological fields and has emerged as a sustainable approach to enhancing agricultural productivity by alleviating various plant stresses. This article reviews innovations in nanobiotechnology and its role in promoting plant growth and enhancing plant resistance/tolerance against biotic and abiotic stresses and the underlying mechanisms. Nanoparticles, synthesized through various approaches (physical, chemical and biological), induce plant resistance against these stresses by strengthening the physical barriers, improving plant photosynthesis and activating plant defense mechanisms. The nanoparticles can also upregulate the expression of stress-related genes by increasing anti-stress compounds and activating the expression of defense-related genes. The unique physico-chemical characteristics of nanoparticles enhance biochemical activity and effectiveness to cause diverse impacts on plants. Molecular mechanisms of nanobiotechnology-induced tolerance to abiotic and biotic stresses have also been highlighted. Further research is needed on efficient synthesis methods, optimization of nanoparticle dosages, application techniques and integration with other technologies, and a better understanding of their fate in agricultural systems.

Acute Pancreatitis Secondary to Pembrolizumab-Induced Hypertriglyceridemia.

Pembrolizumab is a humanized monoclonal antibody targeted against programmed cell death protein 1 (PD-1) receptor of lymphocytes. It is used alone or in combination with many chemotherapy regimens for a wide variety of cancers. It has been reported to cause various side effects including endocrinopathies, colitis, rash, and pneumonitis. Hypertriglyceridemia (HTG) has been recently added to its side effect profile with a possible pathogenic mechanism involving autoantibodies against glycosylphosphatidylinositol-anchored high-density lipoprotein binding protein 1 (GP1HBP1). We are presenting a case of acute pancreatitis secondary to HTG in a cervical cancer patient. HTG was successfully treated with insulin infusion. As the patient's symptoms improved, she was started on the diet. She was discharged on statin and fibrate therapy. We are reporting this case to increase awareness of this rare side effect, inpatient management, and outpatient screening while on immunotherapy.

The mystery of post-cholecystectomy persistent bile leak: a case report.

Laparoscopic cholecystectomy (LC) is commonly performed for benign gallbladder diseases. Biliary leak is the most common complication of bile duct injury following this surgery. We report a case of persistent bile leak following the procedure despite endoscopic and radiological management. A female patient presented to the hepatopancreatobiliary unit of the Bahria International Hospital (Orchard), Lahore, with complaint of persistent bile leakage after laparoscopic cholecystectomy performed elsewhere. She had been investigated in various hospitals but the cause of the persistent bile leak remained a mystery and she was offered surgery. After real time fluoroscopic contrast enhanced imaging, further confirmed by a Computerised Tomography (CT) Scan of the abdomen, it was revealed that the persistent bile leak in the drain was due to iatrogenic injury of the duodenum secondary to percutaneous catheter insertion. The patient was managed non-surgically. She remained stable. This is a rare complication of one of the most common surgical procedures performed in the world.

Primary squamous cell carcinoma of liver. First case report from Pakistan and South Asia.

INTRODUCTION AND IMPORTANCE: Primary squamous cell carcinoma (PSCC) of liver is quite rare and very few cases reported in literature. It has high metastatic rate with poor prognosis. The pathogenesis is unclear, but is generally considered to be correlated with the long-term inflammation or metaplasia of biliary epithelial cells or congenital cyst of the liver. We report here a case of PSCC of liver which mimicked a complex hydatid cyst. CASE PRESENTATION: A 25 years male admitted with right hypochondrium pain associated with fever and yellowish discoloration of eyes for 20 days. He was jaundiced with epigastric tenderness and deranged liver function tests. When thoroughly investigated with ultrasound, CT abdomen and MRI liver, he was found to have a large cystic lesion in right lobe of the liver. He underwent right hepatectomy, peri-cystectomy of the cyst and T-tube placement in common bile duct. Histopathology of the resected sample showed primary squamous cell carcinoma of liver. Patient was discharged after 7 days and died after 6 months due to acute liver failure. CLINICAL DISCUSSION: Because of a very low incidence of hepatic SCC, there is not a single definite therapeutic regime and various different methods of management include surgical resection, generalized chemotherapy, radiotherapy, Hepatic Arterial ChemoEmbolization (HACE) and the combinations of these therapies. CONCLUSION: PSCC is a rare condition of the liver and is associated with other benign liver conditions such as non-parasitic and epidermoid cysts. Histopathology with radiological investigations are needed to diagnose and treat this aggressive tumor before it metastasizes.

A Rare Case of Pancreatic Cancer: Undifferentiated Carcinoma of the Pancreas With Osteoclast-Like Giant Cells.

Ductal adenocarcinoma of the pancreas is the most common pancreatic cancer, but undifferentiated carcinoma of the pancreas with osteoclast-like giant cells (UC-OGCs) is an exceedingly rare tumor. Microscopically, this tumor is characterized by the presence of two different cellular elements, namely, spindle or ovoid mononuclear cells and osteoclast-like giant cells (OGCs). Here, we report a rare case of UC-OGCs in a 79-year-old male with a one-month history of epigastric abdominal pain and unintentional weight loss. A blood workup revealed new-onset type 2 diabetes mellitus, and a computed tomography scan of the abdomen showed acute pancreatitis with a hypodense lesion in the head of the pancreas concerning for malignancy. He underwent an endoscopic ultrasound that also revealed a mass in the head of the pancreas, but no lymphadenopathy was observed. Biopsy was obtained and histopathology revealed UC-OGCs. We present this case to increase awareness of this rare clinical entity in patients presenting with acute-onset pancreatitis.

Silencing of an odorant binding protein (SaveOBP10) involved in the behavioural shift of the wheat aphid Sitobion avenae (Fabricius).

Insects are highly reliant on their active olfactory system in which odorant binding proteins play a role to selectivity and sensitivity during odour perception and processing. This study sets out to determine whether and to which extent the antennal loaded SaveOBP10 in English grain aphid Sitobion avenae, contributes in olfactory processing during host selection. To understand this possible relationship, we purified the SaveOBP10 recombinant protein and performed fluorescence ligand binding tests, molecular docking, RNA interference (RNAi) and behavioural trials. The results showed that SaveOBP10 had strong binding affinities (Ki ≤5 µM) with most of wheat plant volatiles at pH 5.0 as compared to pH 7.4. In Y-tube olfactometer bioassays, the S. avenae was attracted behaviourally towards pentadecane, butylated hydroxytoluene, tetradecane and ß-caryophyllene however repelled by naphthalene. After RNAi of SaveOBP10, the aphid showed nonattraction towards ß-caryophyllene and nonsignificant behavioural response to pentadecane, butylated hydroxytoluene and tetradecane. Furthermore, the three-dimensional structure modelling and molecular docking of SaveOBP10 were performed to the volatiles with high binding abilities. Together these findings indicate that SaveOBP10 can bind more strongly to the volatiles that involved in S. avenae behaviour regulation and possibly will contribute effectively in S. avenae integrated pest management.

In Silico Analysis and Functional Characterization of Antimicrobial and Insecticidal Vicilin from Moth Bean (Vigna aconitifolia (Jacq.) Marechal) Seeds.

Vicilin has nutraceutical potential and different noteworthy medicative health-promoting biotic diversions, and it is remarkable against pathogenic microorganisms and insects. In this study, Vigna aconitifolia vicilin (VacV) has been identified and characterized from the seed of Vigna aconitifolia (Jacq.) Marechal (Moth beans). LC-MS/MS analysis of VacV provided seven random fragmented sequences comprising 238 residues, showing significant homology with already reported Vigna radiata vicilin (VraV). VacV was purified using ammonium sulfate precipitation (60%) followed by size exclusion chromatography on Hi-Load 16/60 Superdex 200 pg column and anion-exchange chromatography (Hi trap Q FF column). Purified VacV showed a major ~50 kDa band and multiple lower bands on 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under both reduced and non-reduced conditions. After all, a three-dimensional molecular structure of VacV was predicted, which showed ß-sheeted molecular conformation similar to crystallographic structure of VraV. All Vicilins from V. aconitifolia and other plants were divided into six sub-groups by phylogenetic analysis, and VacV shared a high degree of similarity with vicilins of Vigna radiata, Pisum sativum, Lupinus albus, Cicer arietinum and Glycine max. Additionally, VacV (20 µg) has significant growth inhibition against different pathogenic bacteria along strong antifungal activity (50 µg). Likewise, VacV (3.0 mg) produced significant growth reduction in Rice Weevil Sitophilus oryzae larvae after 9 days compared with control. Furthermore, by using MMT assay, the cytotoxicity effect of VacV on the growth of HepG2 liver cancerous cells was tested. VacV showed cytotoxicity against the HepG-2 line and the acquired value was 180 µg after 48 h. Finally, we performed molecular docking against caspase-3 protein (PDB ID: 3DEI) for VacV bioactive receptor interface residues. Hence, our results reveal that VacV, has nutraceutical potential and moth beans can be used as a rich resource of functional foods.

First Report of Fusarium Sheath Rot of Rice Caused by Fusarium incarnatum-equiseti Species Complex in the United States.

Fungal diseases, including sheath rot (Sarocladium oryzae), cause significant losses of yield and milling quality of rice (Oryza sativa). In August 2021, symptoms like sheath rot were observed on 20% of rice plants (cv. Presidio) in 1-hectare field in Eagle Lake, Texas. Initial lesions occurred on the upper flag leaf sheaths and were oblong or irregular oval, with gray to light brown centers, and a dark reddish-brown diffuse margin. Lesions enlarged, coalesced, and covered a large area of the sheath. Infection led to panicle rot with kernels turning dark brown. Unlike sheath rot, sheath infection also led to inside culm infection with irregular dark brown lesions. Infected tissue pieces were sterilized with 1% NaOCl for 2 min, followed by 75% ethanol for 30 s, washed in sterile H2O three times, air dried and incubated on PDA at 27℃. Fungal isolates were obtained from 15 diseased plant samples and their singled-spored fungal colonies were whitish, loosely floccose and produced light yellow pigmentation. On carnation leaf agar, macroconidia were slightly curved and tapered at the ends, with 3 to 5 septa, and measured 17.5 to 34.3 × 3.1 to 5.0 µm. Microconidia were ovoid, usually with 0 to 1 septum and were 4.0 to 15.5 × 2.5 to 4.5 µm. Spherical shaped chlamydospores were produced in chain. These morphological characteristics were consistent to those described for Fusarium incarnatum-equiseti species complex (O'Donnell et al. 2009), including F. incarnatum (Wang et al. 2021) and F. equiseti (Avila et al. 2019). For molecular identification, DNA of a representative isolate was extracted and ITS, LSU, and EF1 of the fungus were amplified using the primers of ITS1/ITS4 (Wang et al. 2014), D1/D2 domain region of LSU (Fell et al. 2000), and EF1 (Wang et al. 2014), respectively, and sequenced. The ITS sequence (OL344049) was 99.61% identical to F. incarnatum-equiseti species complex (FD_01692) in Fusarium-ID database and 99.61% identical to F. equiseti (LC514690, KY523100, MW016539) and F. incarnatum (MH979697) in NCBI database. The LSU sequence (OK559512) was 98.77% similar to F. equiseti (MN877913, MN368509) and F. incarnatum (MH877332, MH877326); the EF1 sequence (OK570044) was 99.27% similar to F. equiseti (MK278902) in NCBI database. A phylogenetic analysis based on the concatenated nucleotide sequences grouped this isolate in the F. incarnatum-equiseti species complex clade at 100% bootstrap support. To evaluate pathogenicity, a conidial suspension of 1 x 106 conidia/ml or sterilized water (the controls) was injected into the sheaths and young panicles of three rice plants (cv. Presidio) at boot. Treated plants were maintained in a greenhouse at 25 to 30℃. After 3 weeks, typical symptoms, like those observed in the field, developed on the inoculated plants but not on the controls. The same fungus was consistently re-isolated from the diseased plants. To our knowledge, this is the first report of Fusarium sheath rot caused by F. incarnatum-equiseti species complex in rice in the U. S. F. incarnatum-equiseti species complex has been reported to be associated with panicle infection in wild rice (O. latifolia) in Brazil (Tralamazza et al. 2021). F. incarnatum has also been reported to cause panicle rot in China (Wang et al. 2021). F. proliferatum has been reported to cause Fusarium sheath rot in India (Prabhukarthikeyan et al. 2021) and the U. S. (Cartwright et al. 1995). This research demonstrates the potential of different pathogens being involved in causing sheath rot of rice.

Predicting the impact of environmental factors on citrus canker through multiple regression.

Climatic conditions play a significant role in the development of citrus canker caused by Xanthomonas citri pv. citri (Xcc). Citrus canker is regarded as one of the major threats being faced by citrus industry in citrus growing countries of the world. Climatic factors exert significant impacts on growth stage, host susceptibility, succulence, vigor, survival, multiplication rate, pathogen dispersion, spore penetration rate, and spore germination. Predicting the impacts of climatic factors on these traits could aid in the development of effective management strategies against the disease. This study predicted the impacts of environmental variables, i.e., temperature, relative humidity, rainfall, and wind speed the development of citrus canker through multiple regression. These environmental variables were correlated with the development of canker on thirty (30) citrus varieties during 2017 to 2020. Significant positive correlations were noted among environment variables and disease development modeled through multiple regression model (Y = +24.02 + 0.5585 X1 + 0.2997 X2 + 0.3534 X3 + 3.590 X4 + 1.639 X5). Goodness of fit of the model was signified by coefficient determination value (97.5%). Results revealed the optimum values of environmental variables, i.e., maximum temperature (37°C), minimum temperature (27°C), relative humidity (55%), rainfall (4.7-7.1 mm) and wind speed (8 Km/h), which were conducive for the development of citrus canker. Current study would help researchers in designing better management strategies against citrus canker disease under changing climatic conditions in the future.

First Report of Leaf Spot of Rice Caused by Epicoccum sorghinum in the United States.

Brown spot (Cochliobolus miyabeanus), blast (Magnaporthe oryzae) and stackburn (Alternaria padwickii) are common diseases in rice with similar leaf spot symptoms. In August 2021, a leaf spot disease, with symptoms dissimilar to these diseases, occurred on almost 100% of the leaves and sheaths of rice plants (cv. Presidio) in a 1-hectare field in Eagle Lake, Texas. Lesions started as small dark brown spots on lower leaves and sheaths. The spots enlarged to become round or oval (1.5 to 5.0 mm) spots having round ends with gray centers, dark-brown borders or rings, and slight gold halos. The spots on the sheaths were similar to those on the leaf blades, with lesion size ranging from 2 to 5 mm. Pieces of infected tissue were cut from the margin of necrotic lesions, surface disinfected with 1% NaOCl for 2 min followed by 75% ethanol for 30 s and rinsed with sterile distilled water three times. The tissues were then dried on sterilized filter paper, placed on potato dextrose agar (PDA), and incubated at 25℃ for 7 days. Two isolates (LS36 and LS37) were obtained, and their colonies were initially villose, gray at the center and pale at the margin, and then turned dark gray, with the reverse side becoming scarlet. Chlamydospores were unicellular or multicellular and massively produced in nearly spherical shape (11 to 26 × 10 to 22 µm, n=100). Pycnidia were dark and mostly spheroid (105 to 171 × 76 to 128 µm, n=100). Conidia were unicellular, hyaline, ellipsoidal, with the size of 3.6 to 5.8× 1.9 to 2.8 µm (n=100). These morphological characteristics were similar to those described for Epicoccum sorghinum (Zhou et al. 2018). The rDNA internal transcribed spacer (ITS), rRNA large subunit (LSU), and translation elongation factor 1 alpha (EF1) gene of an representative isolate (LS37) were amplified (Fell et al. 2000; Wang et al. 2014) and sequenced. The ITS sequence (OK189534) of the isolate was 96.95% identical to E. sorghinum (KX758542); the EF1 sequence (OK236518) was 98.37% identical to E. sorghinum (MN461167); and the LSU sequence (OK189535) was 99.29% identical to E. sorghinum (MK817520, MK817521, and MK817522). Rice plants (cv. Presidio) at heading were inoculated with the two isolates individually by placing a drop of conidial suspension of 1 x 106 conidia/ml or a 2-mm PDA plug of 7-day-old cultures on the wounded or unwounded leaves and sheaths (3 sites/leaf or sheath, 3 plants/treatment). The wound was made by penetrating the epidermis using a 0.5-mm-diameter pin. The plants inoculated with sterilized water or PDA-only plugs served as the controls. The treated plants were placed in a dew chamber at 26℃ for 2 days and then transferred in a greenhouse (25 to 30℃). After 5 days, typical symptoms, similar to those observed in the field, developed on all of the inoculated leaves and sheaths, with the wound inoculation inducing more rapid development of symptoms than the unwounded inoculation. No symptoms developed on the controls. The two isolates produced similar symptoms and the fungus was consistently re-isolated from the infected plants and confirmed to be E. sorghinum based on morphological characteristics. The pathogenicity test was repeated twice with similar results. To our knowledge, this is the first report of leaf spot caused by E. sorghinum in rice in the United States. This disease was first reported on rice in China in 2020 (Liu et al. 2020). This research will help identify this new disease from other leaf spot-like diseases and develop management strategies for control of this disease.

First Report of Brown Leaf Spot of Rice Caused by Curvularia hawaiiensis in the United States.

Multiple diseases, including brown spot (Cochliobolus miyabeanus), leaf spot (Epicoccum sorghimum), and blast (Magnaporthe oryzae), can cause spot-like symptoms on the leaves of rice. In July 2021, a disease showing symptoms like brown spot was observed in an 8-hectare field of rice, with disease incidence of >30%, in Beaumont, Texas. Lesions started as small pinhead-size blackish spots on leaf tips or from the edges of leaf blades. The spots enlarged to become irregular (most) or oval brown spots with a slight chlorotic halo. Diseased leaves were collected, washed in running tap water and cut into small pieces. Pieces of the tissue were surface sterilized with 1%NaOCl for 2 min followed by 75% ethanol for 30 s and then washed in sterile distilled water three times with each time lasting for 1 min. The disinfected tissue pieces were air dried, placed on potato dextrose agar (PDA) medium and incubated at 25℃. Initially fungal colonies were hairy in texture with light dark brown center and whitish edge and dark brown pigmentation at the reverse side. Mature colonies turned to black in the center and dark brown toward the edge, with black at the reverse side after 2 or more weeks of incubation. Conidia were oval to narrowly oblong, rounded at the ends, with 2 to 6 distoseptate, and 15 to 35 × 6 to 10 µm in size. These morphological characteristics were similar to those described for Curvularia hawaiiensis (Aslam et al. 2019; Ellis 1971; Kusai et al. 2015). For molecular identification, DNA was extracted and the two different rRNA regions internal transcribed spacer (ITS) and large subunit (LSU), and the two genes RNA Polymerase II (RPB1) and translation elongation factor 1 alpha (EF1) of the fungus were amplified using the primers of ITS1/ITS4 (Wang et al. 2014), D1/D2 domain region of LSU (Fell et al. 200), and RPB1 and EF1 (Wang et al. 2014), respectively, and sequenced. The ITS sequence (OK397200) was 98.27% identical to C. hawaiiensis (KP131943); the EF1 sequence (OK492159) was 99.78% identical to C. hawaiiensis (KC503942); the LSU sequence (OK397295) was 98.96% identical to multiple C. hawaiiensis (MN055715, MH160813, MH875853, etc.); the RPB1 sequence (OK492160) was 97.41% identical to C. hawaiiensis (JN992363). To evaluate pathogenicity, three rice plants (cv. Presidio) at the 3-leaf stage were spray inoculated with a conidial suspension of 1 x 106 conidia/ml. Another set of three plants that were sprayed with sterilized distilled water served as the controls. Treated plants were maintained in a greenhouse with temperature ranging from 25 to 30℃. After 2 weeks, typical symptoms, like those observed in the field, developed on the inoculated plants while no symptoms developed on the control plants. The same fungus was consistently re-isolated from the diseased plants. The pathogenicity test was conducted three times with similar results. To our knowledge, this is the first report of brown leaf spot caused by C. hawaiiensis in rice in the United States. Curvularia species are frequently associated with rice grain and cause blackish discoloration symptoms on grain kernels. Recently, however, C. hawaiiensis has also been reported to cause brown leaf spot in Malaysia (Kusai et al. 2015) and Pakistan (Aslam et al. 2019). This research will help identify this disease from other leaf spot-like diseases and develop effective management strategies.

Clinical outcomes of hepato-pancreato-biliary surgical patients during COVID-19 pandemic.

OBJECTIVE: To determine the outcome of hepato-pancreato-biliary patients who were operated upon during the pandemic under a safety protocol devised to minimise the risk of coronavirus disease-2019 infection in patients and staff during the perioperative period. METHODS: The retrospective study was conducted at the Hepato-Pancreatico-Biliary (HPB) Unit of Bahria International Hospital Orchard, Lahore, Pakistan, in February 2021, and comprised data of all patients who were discussed in the multidisciplinary meeting of the Unit between May 1 and December 31, 2020. The coronavirus disease-2019 screening protocol was a negative polymerase chain reaction test just before admission and a second negative test 24-48 hours pre-surgery. All patients had computed tomography scan of chest to rule out atypical pneumonia due to coronavirus disease-2019 infection. Surgery was deferred for positive patients for at least 2 weeks until their test was negative. Surgeries were carried out with full personal protective equipment. Further testing was carried out if clinically indicated. Data was collected of 30-day coronavirus disease-2019-related morbidity and mortality. Data was analysed using SPSS 20. RESULTS: Of the 44 patients, 29(65.9%) were males and 15(34.1%) were females. Overall, 32(72.7%) patients were aged >40 years, 8(18.2%) were aged 20-40 years and 4(9%) were aged <20 years. Of the total, 7(15.09%) patients were found to be positive for coronavirus disease-2019 during preoperative screening. Among them, 5(71.4%) had successful surgery post-recovery, 1(14.3%) died without surgery due to pulmonary complications related to coronavirus disease-2019 and 1(14.3%) patient was lost to follow-up. Among the 42(95.5%) patients who underwent any procedure, 2(4.7%) patients turned positive for coronavirus disease-2019 in the postoperative period. There was 1(2.4%) mortality in the 30-day post-operative period which was not related to coronavirus disease-2019. Complications were seen in 5(11.90%) patients. Three (9.3%) out of 32 staff members were found to be asymptomatic carriers. CONCLUSIONS: Hepato-pancreatico-biliary surgery could be safely carried out during coronavirus disease-2019 pandemic if strict safety protocols were followed.

Citrus Canker Pathogen, Its Mechanism of Infection, Eradication, and Impacts.

Citrus canker is a ravaging bacterial disease threatening citrus crops. Its major types are Asiatic Canker, Cancrosis B, and Cancrosis C, caused by Xanthomonas citri pv. citri (Xcc), Xanthomonas citri pv. aurantifolii pathotype-B (XauB), and pathotype-C (XauC), respectively. The bacterium enters its host through stomata and wounds, from which it invades the intercellular spaces in the apoplast. It produces erumpent corky necrotic lesions often surrounded by a chlorotic halo on the leaves, young stems, and fruits, which causes dark spots, defoliation, reduced photosynthetic rate, rupture of leaf epidermis, dieback, and premature fruit drop in severe cases. Its main pathogenicity determinant gene is pthA, whose variants are present in all citrus canker-causing pathogens. Countries where citrus canker is not endemic adopt different methods to prevent the introduction of the pathogen into the region, eradicate the pathogen, and minimize its dissemination, whereas endemic regions require an integrated management program to control the disease. The main aim of the present manuscript is to shed light on the pathogen profile, its mechanism of infection, and fruitful strategies for disease management. Although an adequate method to completely eradicate citrus canker has not been introduced so far, many new methods are under research to abate the disease.

Health-Related Quality of Life of Tuberculosis Patients and the Role of Socioeconomic Factors: A Mixed-Method Study.

This study aims to assess tuberculosis (TB) patients' health-related quality of life (HRQoL) and to determine the relationship between HRQoL and the socioeconomic and health characteristics of patients. A mixed-method approach of explanatory sequential design is used in this study. Data were collected at 11 TB centers and two private clinics in Khyber Pakhtunkhwa, Pakistan. Quantitative data were collected through a standardized questionnaire (SF-36) of RAND organization from 269 pulmonary TB patients. Qualitative data were collected through 20 in-depth interviews, 15 Key Informant interviews, and a focus group discussion. The maximum score of SF-36 is 100, however, different aspects of HRQoL indicated that patients scored lowest in the role limitation physical (10.3) and emotional (11.2) categories, while the highest remained in the mental health (45.3) category. Patients of higher age, being female, low household income, and comorbidity decreased the HRQoL. Qualitative findings show that TB affected the patients' social functioning, vitality, and emotions. Moreover, poverty and low income, people living in Kacha Houses, family support, and the hate from family members and community are related to low HRQoL. In summary, TB has affected the Patients' HRQoL. TB program managers should pay attention to the nonmedication aspects of TB management. Financial support should be extended TB patients. This study calls for the urgent attention of the National Tuberculosis Program and policymakers, for an increased focus on patients' welfare programs.

Two Infectious Agents Causing Hemophagocytic Lymphohistiocytosis.

Hemophagocytic lymphohistiocytosis (HLH) is a life-threatening immune activation syndrome that should be recognized earlier for effective treatment. Adults usually have secondary HLH. An uncommon cause of secondary HLH is AIDS and simultaneous opportunistic infections. Acute human immunodeficiency virus (HIV) and opportunistic infections are also independent causes of HLH, so the presence of both should raise suspicion, especially if patients fulfill the criteria. HLH secondary to severe babesiosis is a rare entity as well. Some patients might not meet the full criteria of HLH on presentation, especially when some specific lab test results are still pending. A delay in diagnosis can happen in those cases. Here, we present two cases. The first case is of a 35-year-old homosexual male who presented with constitutional symptoms of one-week duration. He was diagnosed and started on the treatment of HIV. His fever was not resolving and further investigations led to a diagnosis of disseminated histoplasma infection. The patient fulfilled the criteria of HLH as well. Prompt therapy resulted in the improvement of clinical and laboratory parameters. The second case is of a 72-year-old female presenting with fever. A diagnosis of severe babesiosis and secondary HLH was made. Treatment of babesia resulted in the improvement of clinical and biochemical parameters.