Phytochemical screening and biological activities of Bromus pectinatus Thunb.

The present research study investigates the phytochemical and pharmacological importance of Bromus pectinatus. Qualitative phytochemical analysis of this plant was carried out to use standard method for the presence of various bioactive constituents. Results showed the ethanolic extract contain natural product such as steroids, alkaloids, tannins, coumarin, saponins, flavonoids and phenols. These compounds play a key role to reducing various disease and microbial inhibition. The ethanolic extract also showed the antimicrobial and antifugal activity against different pathogenic bacterial strains e.g Escherichia coli, Micrococus leutus, Protus vulgarus, and Kelebsela pneumona and three fungal strains Aspergillus fumigatus, Aspergillus flavous, Aspergillus niger. The antioxidant assay was performed as % inhibition of DPPH (1, 1-diphenyl-2-picryl-hydrazyl) free radicals. The plant extract has more antioxidant activity as compared to ascorbic acid. The maximum concentration (800µg/ml) is the most effective of all. The plant extract showed the high cytotoxicity activity against Brine shrimp. Moreover, the plant extract exhibited allelopathic effect on different growth parameters of wheat plant mostly at higher concentration. These results indicate that the BPEE have a potential broad-spectrum antimicrobial, cytotoxic, antioxidant and phytotoxic activity due to the presence of bioactive compounds.

Microvirga pakistanensis sp. nov., a novel bacterium isolated from desert soil of Cholistan, Pakistan.

A Gram-negative, non-spore-forming, non-pigmented, strictly aerobic and non-motile short rod bacterium, designated NCCP-1258T, was isolated from Cholistan desert soil, Bahawalpur, Pakistan. Growth of strain NCCP-1258T was observed at pH range 6.5-9.5 (optimum 7.5-8.5) and temperature range 20-45 °C (optimum 40 °C), and it tolerated 0-2 % NaCl (optimum 0.5 %, w/v). Phylogenetic analysis based on 16S rRNA gene sequence comparison revealed that strain NCCP-1258T belongs to genus Microvirga and is most closely related to Microvirga lotononidis (98.0 %), Microvirga vignae (97.4 %), Microvirga lupini (97.2 %), Microvirga zambiensis (97.2 %) and Microvirga flocculans (97.1 %). Analysis of the concatenated sequences of four housekeeping gene loci (dnaK, gyrB, recA and rpoB) also confirmed the placement of strain NCCP-1258T within the genus Microvirga. DNA-DNA relatedness values of NCCP-1258T with above-mentioned type strains were less than 42 %. The DNA G+C content of strain NCCP-1258T was 64.3 mol%. Chemotaxonomic data (predominant menaquinone system was Q-10; major fatty acids were C16:0, C18:1 ω7c and C19:0 cyclo ω8c; the polar lipid profile contained diphosphatidylglycerol, phosphatidylcholine, phosphatidyl dimethyl ethanolamine and phosphatidyl ethanolamine) also supported the affiliation of strain NCCP-1258T to the genus Microvirga. On the basis of physiological and biochemical characteristics, phylogenetic analyses and DNA-DNA relatedness, strain NCCP-1258T can be distinguished from the closely related taxa and thus represents a novel species of the genus Microvirga, for which the name Microvirga pakistanensis sp. nov. is proposed with the type strain NCCP-1258T (=CGMCC 1.15074T = KCTC 42496T).

Nocardioides pakistanensis sp. nov., isolated from a hot water spring of Tatta Pani in Pakistan.

A Gram-staining positive, non-spore forming, non-pigmented and non-motile bacterium, designated as NCCP-1340(T), was isolated from a hot water spring, Tatta Pani, Pakistan. Cells of strain NCCP-1340(T) were observed to be aerobic, rod shaped, catalase and urease positive but H2S production and oxidase negative. Growth was observed at pH 6.0-8.0 (optimum pH 7.0) and at 20-40 °C (optimum 37 °C). The strain could tolerate 0-8 % NaCl (optimum 2 %, w/v). Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain NCCP-1340(T) belongs to the genus Nocardioides and is closely related to Nocardioides iriomotensis JCM 17985(T) (96.8 %), Nocardioides daedukensis KCTC 19601(T) (96.6 %), Nocardioides jensenii KCTC 9134(T) (96.1 %) and Nocardioides daejeonensis KCTC 19772(T) (96.1 %). The DNA-DNA relatedness values of strain NCCP-1340(T) with N. iriomotensis JCM 17985(T), N. daedukensis KCTC 19601(T) and N. jensenii KCTC 9134(T) were found to be less than 53 %. The DNA G+C content of strain NCCP-1340(T) was determined to be 71.8 mol  %. The affiliation of strain NCCP-1340(T) to the genus Nocardioides was further supported by chemotaxonomic data which showed the presence of MK-8(H4) as major menaquinone system; iso-C16:0, C17:0, C16:0 10-methyl, iso-C15:0 and C 15:0 as major cellular fatty acids; and diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and unidentified glycolipids and polar lipids in the polar lipids profile. The cell wall peptidoglycan contained LL-diaminopimelic acid as the diagnostic amino acid. On the basis of physiological and biochemical characteristics and the phylogenetic analyses, strain NCCP-1340(T) can be distinguished from the closely related taxa and thus represents a novel species of the genus Nocardioides, for which the name Nocardioides pakistanensis sp. nov. is proposed with the type strain NCCP-1340(T) (= DSM 29942(T) = JCM 30630(T)).

A heavy metal tolerant novel bacterium, Bacillus malikii sp. nov., isolated from tannery effluent wastewater.

The taxonomic position of a Gram-stain positive and heavy metal tolerant bacterium, designated strain NCCP-662(T), was investigated by polyphasic characterisation. Cells of strain NCCP-662(T) were observed to be rod to filamentous shaped, motile and strictly aerobic, and to grow at 10-50 °C (optimum 30-37 °C) and at pH range of 6-10 (optimum pH 7-8). The strain was found to be able to tolerate 0-12 % NaCl (w/v) and heavy metals (Cr 1200 ppm, Pb 1800 ppm and Cu 1200 ppm) in tryptic soya agar medium. The phylogenetic analysis based on the 16S rRNA gene sequence of strain NCCP-662(T) showed that it belongs to the genus Bacillus and showed high sequence similarity (98.2 and 98.0 %, respectively) with the type strains of Bacillus niabensis 4T19(T) and Bacillus halosaccharovorans E33(T). The chemotaxonomic data showed that the major quinone is MK-7; the predominant cellular fatty acids are anteiso-C15 :0, iso-C14:0, iso-C16:0 and C16:0 and iso-C15:0; the major polar lipids are diphosphatidylglycerol, phosphatidylglycerol along with several unidentified glycolipids, phospholipids and polar lipids. The DNA G+C content was determined to be 36.9 mol%. These data also support the affiliation of strain NCCP-662(T) with the genus Bacillus. The level of DNA-DNA relatedness between strain NCCP-662(T) and B. niabensis JCM 16399(T) was 20.5 ± 0.5 %. On the basis of physiological and biochemical characteristics, phylogenetic analyses and DNA-DNA hybridization data, strain NCCP-662(T) can be clearly differentiated from the validly named Bacillus species and thus represents a new species, for which the name Bacillus malikii sp. nov. is proposed with the type strain NCCP-662(T) (= LMG 28369(T) = DSM 29005(T) = JCM 30192(T)).

A heavy-metal tolerant novel bacterium, Alcaligenes pakistanensis sp. nov., isolated from industrial effluent in Pakistan.

Two strains, NCCP-650(T) and NCCP-667, were isolated from industrial effluent and their taxonomic positions were investigated using a polyphasic taxonomic approach. The strains were found to be Gram-stain negative, strictly aerobic, motile short rods, which are tolerant to heavy-metals (Cr(+2), As(+2), Pb(+2) and Cu(+2)). Cells were observed to grow at a temperature range of 10-37 °C (optimal 25-33 °C), pH range of 5.5-10.0 (optimal 6.5-7.5) and can tolerate 0-7 % NaCl (w/v) (optimum 0-1 %) in tryptic soya agar medium. Sequencing of the 16S rRNA gene and two housekeeping genes, gyrB and nirK, of the isolated strains revealed that both strains belong to the Betaproteobacteria showing highest sequence similarities with members of the genus Alcaligenes. The chemotaxonomic data [major quinones as Q-8; predominant cellular fatty acids as summed features 3 (C16 :1 ω7c/iso-C15 :0 2OH) and C16:0 followed by Summed features 2 (iso-C16 :1 I/C14 :0 3OH), C17:0 Cyclo and C18:1 ω7c; major polar lipids as diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and one unidentified aminolipid] also supported the affiliation of the isolated strains with the genus Alcaligenes. DNA-DNA hybridizations between the two strains and with closely related type strains of species of the genus Alcaligenes confirmed that both isolates belong to a single novel species within the genus Alcaligenes. On the basis of phylogenetic analyses, physiological, biochemical characteristics and DNA-DNA hybridization, the isolated strains can be differentiated from established Alcaligenes species and thus represent a novel species, for which the name Alcaligenes pakistanensis sp. nov. is proposed with the type strain NCCP-650(T) (=LMG 28368(T) = KCTC42083(T) = JCM 30216(T)).

Kushneria pakistanensis sp. nov., a novel moderately halophilic bacterium isolated from rhizosphere of a plant (Saccharum spontaneum) growing in salt mines of the Karak area in Pakistan.

The taxonomic position of a Gram-stain negative, moderately halophilic bacterium, designated NCCP-934(T), was investigated using polyphasic taxonomic approach. The strain NCCP-934(T) was isolated from rhizosphere of a plant (Saccharum spontaneum, family Poaceae) growing in salt mines area in the Karak district of Khyber Pakhtunkhwa Province, Pakistan. Cells of strain NCCP-934(T) are rod shaped and motile. The bacterium is strictly aerobic, can grow at a temperature range of 10-40 °C (optimum at 30-33 °C) and in a pH range of 6.0-10.5 (optimum pH 7.0-9.0). The strain can tolerate 1-30 % (w/v) NaCl (optimal growth occurs in the presence of approximately 3-9 % NaCl). The phylogenetic analysis based on the 16S rRNA gene sequence, showed that strain NCCP-934(T) belongs to the genus Kushneria with the highest sequence similarity to K. marisflavi SW32(T) (98.9 %), K. indalinina CG2.1(T) (98.7 %), K. avicenniae MW2a(T) (98.4 %) and less than 97 % similarity with other related species (94.7 % with the type species of the genus, K. aurantia A10(T)). DNA-DNA relatedness between strain NCCP-934(T) and the type strains of the closely related species was lower than 18 %. The chemotaxonomic data (major respiratory quinone, Q9; predominant fatty acids, C18:1 ω7c and C16:0 followed by C12:0 3-OH and Summed features 3 (C16:1 ω7c/iso-C15:0 2-OH); major polar lipids, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmonomethylethanol, phosphatidylserine, phosphatidylinositol and three polar lipid of unknown structure) supported the affiliation of strain NCCP-934(T) within the genus Kushneria. The DNA G+C content of strain NCCP-934(T) was 59.2 mol%. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain NCCP-934(T) can be distinguished from the closely related taxa and thus represents a novel species in the genus Kushneria, for which the name Kushneria pakistanensis sp. nov. is proposed, with the type strain NCCP-934(T) (=LMG 28525(T) = KCTC 42082(T) = JCM 18802(T)).

Deinococcus citri sp. nov., isolated from citrus leaf canker lesions.

A Gram-stain-positive, strictly aerobic, non-motile, coccoid bacterium, designated NCCP-154(T), was isolated from citrus leaf canker lesions and was subjected to a polyphasic taxonomic study. Strain NCCP-154(T) grew at 10-37 °C (optimum 30 °C) and at pH 7.0-8.0 (optimum pH 7.0). The novel strain exhibited tolerance of UV irradiation (>1000 J m(-2)). Based on 16S rRNA gene sequence analysis, strain NCCP-154(T) showed the highest similarity to Deinococcus gobiensis CGMCC 1.7299(T) (98.8 %), and less than 94 % similarity to other closely related taxa. The chemotaxonomic data [major menaquinone, MK-8; cell-wall peptidoglycan type, A3ß (Orn-Gly2); major fatty acids, summed feature 3 (C16 : 1ω7c/iso-C15 : 0 2-OH; 35.3 %) followed by C16 : 0 (12.7 %), iso-C17 : 1ω9c (9.2 %), C17 : 1ω8c (7.4 %) and iso-C17 : 0 (6.9 %); major polar lipids made up of several unidentified phosphoglycolipids and glycolipids and an aminophospholipid, and mannose as the predominant whole-cell sugar] also supported the affiliation of strain NCCP-154(T) to the genus Deinococcus. The level of DNA-DNA relatedness between strain NCCP-154(T) and D. gobiensis JCM 16679(T) was 63.3±3.7 %. The DNA G+C content of strain NCCP-154(T) was 70.0 mol%. Based on the phylogenetic analyses, DNA-DNA hybridization and physiological and biochemical characteristics, strain NCCP-154(T) can be differentiated from species with validly published names. Therefore, it represents a novel species of the genus Deinococcus. The name Deinococcus citri sp. nov. is proposed, with the type strain NCCP-154(T) ( = JCM 19024(T) = DSM 24791(T) = KCTC 13793(T)).

Cellulomonas pakistanensis sp. nov., a moderately halotolerant Actinobacteria.

A rod-shaped, motile, facultatively anaerobic and moderately halotolerant plant-growth-promoting actinobacterial strain, designated NCCP-11(T), was isolated from paddy grains. To delineate its taxonomic position, the strain was subjected to a polyphasic characterization. Cells of strain NCCP-11(T) grew at 10-37 °C (optimum 28-32 °C), at pH 6-9 (optimum pH 7) and in 0-12% (w/v) NaCl (optimum 1-2%) in broth medium. Based on 16S rRNA gene sequence analysis, strain NCCP-11(T) showed highest similarity to the type strains of Cellulomonas hominis (98.99%) and Cellulomonas denverensis (98.09 %) and less than 97 % with other closely related taxa. The chemotaxonomic data [major menaquinone: MK-9(H4); cell-wall peptidoglycan: type A4ß; major fatty acids: anteiso-C15 : 0, C16 : 0, C14 : 0 and anteiso-C17 : 0; major polar lipids: diphosphatidylglycerol, phosphatidylinositol, phosphatidylinositolmannosides and two unknown polar lipids] also supported the affiliation of strain NCCP-11(T) to the genus Cellulomonas. The level of DNA-DNA relatedness between strain NCCP-11(T) and the two type strains mentioned above was less than 42.7%. On the basis of DNA-DNA relatedness, physiological and biochemical characteristics and phylogenetic position, strain NCCP-11(T) can be differentiated from species of the genus Cellulomonas with validly published names and thus represents a novel species, for which the name Cellulomonas pakistanensis sp. nov. is proposed. The type strain is NCCP-11(T) ( = DSM 24792(T) = JCM 18755(T) = KCTC 19798(T)).

Bacillus pakistanensis sp. nov., a halotolerant bacterium isolated from salt mines of the Karak Area in Pakistan.

A rod shaped, non-motile, endospore forming, Gram-stain positive and moderately halotolerant strain, designated as NCCP-168(T), was isolated from salt mines sampled in the Karak district of Khyber Pakhtunkhwa Province in Pakistan. To delineate its taxonomic position, the strain was subjected to polyphasic characterization. Cells of strain NCCP-168(T) can grow at 10-40 (○)C (optimum at 30-35 (○)C), in a pH range of 5.0-9.0 (optimum at pH 8.0) and in 0-17 % (w/v) NaCl on agar medium. The phylogenetic analysis based on the 16S rRNA gene sequence showed that strain NCCP-168(T) belongs to the genus Bacillus with the highest similarity to Bacillus seohaeanensis BH724(T) (97.1 %), and less than 97 % similarity with other closely related taxa (95.6 % with B. subtilis subsp. subtilis NCIB3610(T)). DNA-DNA relatedness between strain NCCP-168(T) and the type strains of closely related species was lower than 30 %. Chemotaxonomic data (major menaquinone, MK-7; cell wall peptidoglycan type, A1γ [meso-diaminopimelic acid]; major fatty acids, iso-C15:0 29.9 %, anteiso-C15:0 29.3 %, iso-C16:0 11.4 %, iso-C14:0 8.9 % and anteiso-C17:0 7.0 %; major polar lipids, diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine) support the affiliation of strain NCCP-168(T) with genus Bacillus. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain NCCP-168(T) can be distinguished from the closely related taxa and thus represents a novel species in the genus Bacillus, for which the name Bacillus pakistanensis sp. nov. is proposed, with the type strain NCCP-168(T) (= KCTC 13786(T) = DSM 24834(T) = JCM 18975(T)).

Probing the microbial diversity and probiotic candidates from Pakistani foods: isolation, characterization, and functional profiling.

Probiotics represent beneficial living microorganisms that confer physiological, nutritional, and functional advantages to human health, holding significant potential for development of functional foods. This research aimed to isolate, identify, and characterize potential probiotic bacterial strains sourced from fermented and non-fermented foods from Pakistan. A total of 341 bacterial strains were isolated from diverse food samples (81) collected from various regions of Pakistan. Strains were identified using 16S rRNA gene sequencing and phylogenetic analysis. The identified strains belonged to genera Bacillus, Staphylococcus, Microbacterium, Shigella, Micrococcus, Enterococcus, Sporosarcina, Paenibacillus, Limosilactobacillus, Kosakonia, Dietzia, Leclercia, Lacticaseibacillus, Levilactobacillus, Kluyvera, Providencia, Enterobacter, Neisseria, Streptococcus, Acinetobacter, Corynebacterium, Pantoea, Mammaliicoccus, Pseudomonas, Burkholderia, and Alkalihalobacillus. Selected strains were chosen for probiotic assessment, employing existing literature as a guideline. Among these selections, six strains exhibited hemolytic activity, and seven strains displayed resistance to multiple antibiotics, prompting their exclusion from subsequent evaluations. The remaining strains demonstrated auto-aggregation capacities spanning 3.39-79.7%, and displayed coaggregation capabilities with reported food-borne pathogens. Furthermore, nine strains exhibited antimicrobial properties against food-borne pathogens. The assessment encompassed diverse characteristics such as cell surface hydrophobicity, survival rates under varying conditions, cholesterol reduction ability, casein digestion capability, and antioxidant activity. Phylogenomic analysis, digital-DNA DNA hybridization (digi-DDH), and average nucleotide identity (ANI) calculations unveiled novel species potentially belonging to the genera Sporosarcina and Dietzia. Based on these findings, we advocate for the consideration of Staphylococcus cohnii subsp. cohnii NCCP-2414, Lacticaseibacillus rhamnosus NCCP-2569 and Levilactobacillus brevis NCCP-2574 as prime probiotic candidates with the potential for integration into formulation of functional foods. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03903-6.

Growth improvement of wheat (Triticum aestivum) and zinc biofortification using potent zinc-solubilizing bacteria.

Zinc (Zn) is an indispensable element for proper plant growth. A sizeable proportion of the inorganic Zn that is added to soil undergoes a transformation into an insoluble form. Zinc-solubilizing bacteria (ZSB) have the potential to transform the insoluble Zn into plant-accessible forms and are thus promising alternatives for Zn supplementation. The current research was aimed at investigating the Zn solubilization potential of indigenous bacterial strains and to evaluate their impact on wheat growth and Zn biofortification. A number of experiments were conducted at the National Agriculture Research Center (NARC), Islamabad, during 2020-21. A total of 69 strains were assessed for their Zn-solubilizing ability against two insoluble Zn sources (ZnO and ZnCO3) using plate assay techniques. During the qualitative assay, the solubilization index and solubilization efficiency were measured. The qualitatively selected Zn-solubilizing bacterial strains were further tested quantitatively using broth culture for Zn and phosphorus (P) solubility. Tricalcium phosphate was used as insoluble source of P. The results showed that broth culture pH was negatively correlated with Zn solubilization, i.e., ZnO (r2 = 0.88) and ZnCO3 (r2 = 0.96). Ten novel promising strains, i.e., Pantoea sp. NCCP-525, Klebsiella sp. NCCP-607, Brevibacterium sp. NCCP-622, Klebsiella sp. NCCP-623, Acinetobacter sp. NCCP-644, Alcaligenes sp. NCCP-650, Citrobacter sp. NCCP-668, Exiguobacterium sp. NCCP-673, Raoultella sp. NCCP-675, and Acinetobacter sp. NCCP-680, were selected from the ecology of Pakistan for further experimentation on wheat crop based on plant growth-promoting rhizobacteria (PGPR) traits, i.e., solubilization of Zn and P in addition to being positive for nifH and acdS genes. Before evaluating the bacterial strains for plant growth potential, a control experiment was also conducted to determine the highest critical Zn level from ZnO to wheat growth using different Zn levels (0.1, 0.05, 0.01, 0.005, and 0.001% Zn) against two wheat varieties (Wadaan-17 and Zincol-16) in sand culture under glasshouse conditions. Zinc-free Hoagland nutrients solution was used to irrigate the wheat plants. As a result, 50 mg kg-1 of Zn from ZnO was identified as the highest critical level for wheat growth. Using the critical level (50 mg kg-1 of Zn), the selected ZSB strains were inoculated alone and in consortium to the seed of wheat, with and without the use of ZnO, in sterilized sand culture. The ZSB inoculation in consortium without ZnO resulted in improved shoot length (14%), shoot fresh weight (34%), and shoot dry weight (37%); with ZnO root length (116%), it saw root fresh weight (435%), root dry weight (435%), and Zn content in the shoot (1177%) as compared to the control. Wadaan-17 performed better on growth attributes, while Zincol-16 had 5% more shoot Zn concentration. The present study concluded that the selected bacterial strains show the potential to act as ZSB and are highly efficient bio-inoculants to combat Zn deficiency, and the inoculation of these strains in consortium performed better in terms of growth and Zn solubility for wheat as compared to individual inoculation. The study further concluded that 50 mg kg-1 Zn from ZnO had no negative impact on wheat growth; however, higher concentrations hampered wheat growth.

Unraveling the draft genome and phylogenomic analysis of a multidrug-resistant Planococcus sp. NCCP-2050T: a promising novel bacteria from Pakistan.

Planococcus is a genus of Gram-positive bacteria known for potential industrial and agricultural applications. Here, we report the first draft genome sequence and phylogenomic analysis of a CRISPR-carrying, multidrug-resistant, novel candidate Planococcus sp. NCCP-2050T isolated from agricultural soil in Pakistan. The strain NCCP-2050T exhibited significant resistance to various classes of antibiotics, including fluoroquinolones (i.e., ciprofloxacin, levofloxacin, ofloxacin, moxifloxacin, and bacitracin), cephalosporins (cefotaxime, ceftazidime, cefoperazone), rifamycins (rifampicin), macrolides (erythromycin), and glycopeptides (vancomycin). Planococcus sp. NCCP-2050T consists of genome size of 3,463,905 bp, comprised of 3639 annotated genes, including 82 carbohydrate-active enzyme genes and 39 secondary metabolite genes. The genome also contained 80 antibiotic resistance, 162 virulence, and 305 pathogen-host interaction genes along with two CRISPR arrays. Based on phylogenomic analysis, digital DNA-DNA hybridization, and average nucleotide identity values (i.e., 35.4 and 88.5%, respectively) it was suggested that strain NCCP-2050T might represent a potential new species within the genus Planococcus. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03748-z.

Frequency of skeletal dysplasia in children with short stature presenting to endocrine clinic: An observational study.

Objective: To determine the frequency of skeletal dysplasia in children with short stature presenting to the endocrine clinic of a tertiary care hospital. Methods: This descriptive cross-sectional study was performed in the Outpatient Department of Endocrinology of National Institute of Child Health, Karachi, for 6 months of duration. A total of 200 children coming to endocrine OPD of NICH of either gender, having the age less than 14 years and height more than -2.5 SD below the mean (<3rd percentile), and growth failure (<4 cm/yr) were enrolled. A complete general physical examination including height, weight, fronto-occipital circumference (FOC), arm span, and U/L (upper/lower) segment ratio (using SI units and SDS) was performed. Results: Out of 200 children with short stature, skeletal dysplasia was diagnosed in 23 (11.5%) children with the mean age of 4.7 (±3.7) years. Proportion of skeletal dysplasia among short stature was high in females. Out of 75 girls, skeletal dysplasia was diagnosed in 10 (13.3%) girls, while out of 125 boys, skeletal dysplasia was diagnosed in 13 (10.4%) boys, whereas when we see proportion among skeletal dysplasia out of 23 children of skeletal dysplasia, 13 (56.5%) were boys, while 10 (43.5%) were girls. Conclusion: In this study, skeletal dysplasia was diagnosed in 11.5% children with short stature with the mean age of 4.7 years. It is concluded that the frequency of skeletal dysplasia in this institute is fairly high.

An Observational Study to Evaluate the Effectiveness of Rivaroxaban in the Management of Cerebral Venous Sinus Thrombosis.

Background and objectives Cerebral venous sinus thrombosis (CVST) is a relatively rare and underdiagnosed type of stroke. Rivaroxaban is licensed for venous thromboembolism in patients opting for elective knee and hip replacement surgeries, preventing pulmonary embolism and precluding stroke and systemic embolism in sufferers of non-valvular atrial fibrillation. Beneficial outcomes depicting the efficacious role of rivaroxaban in treating CVST are under study. Materials and methods We performed a prospective observational study in patients diagnosed with CVST in the medicine or neurology unit of a tertiary care hospital in Karachi, Pakistan, between January 2019 and December 2019. The diagnosis of CVST was made by magnetic resonance venography (MRV) in all the cases. Follow-up visits were scheduled at three months and six months, and the occurrence of thrombotic events or bleeding complications was recorded. Follow-up was done by magnetic resonance imaging at three and six months to assess vessel recanalization. Excellent outcome was defined as a modified Rankin Scale (mRS) of 0 or 1. A total of 31 patients were meeting the inclusion criteria and were inducted into the study after informed consent. Results The mean age of the study population was 35.11 ± 8.96 years with 71% females and 29% males. The most prevalent etiology was the pregnancy/postpartum period (52%) followed by antiphospholipid syndrome (23%). The frequent clinical manifestations were headache (84%) followed by vomiting (38%), altered level of consciousness (35%), focal deficit/limb weakness (32%), aphasia (29%), blurring of vision (26%), and seizures (23%). Radiological studies showed that the vessels chiefly occluded in our study were superior sagittal sinus (29%), transverse sinus (23%), sigmoid sinus (16%), jugular vein (9%), and cortical veins (3%). Common features on magnetic resonance imaging (MRI) were cerebral edema (45%), hemorrhage (39%), infarct incidence (32%), and raised intracranial pressure (26%). Clinical outcomes showed 55% of patients had partial recanalization and 39% had complete recanalization after a period of six months of the administration of rivaroxaban. Ninety-three percent (93%) of sufferers recovered excellently according to mRS and only 3% developed recurrent CVST within a span of six months. The frequency of thrombotic events and bleeding complications were reported in 6% of patients, respectively, while mortality reported was also 6%. Conclusion Rivaroxaban has shown promising results in the management of our CVST patients, hence, it further warrants randomized controlled trials of rivaroxaban against conventional treatments to prove its significant role.

Frequency of Comorbidities in Admitting COVID-19 Pneumonia Patients in a Tertiary Care Setup: An Observational Study.

Background The novel coronavirus disease 2019 (COVID-19) is a highly infectious and pandemic disease with a variable mode of action. Patients with underlying illnesses such as diabetes, hypertension, and other diseases are more prone to infection. An understanding of the different comorbidities that place patients at the highest risk of COVID-19 pneumonia and other fatal complications associated with COVID-19 is necessary for healthcare professionals. This study aimed to determine the frequency of different comorbid illnesses among COVID-19 patients admitted to a tertiary care hospital in Karachi, Pakistan. Methodology All patients diagnosed with COVID-19 who required admission for the care of their symptoms were included in this observational, cross-sectional study conducted from May 1 to July 30, 2020. The patients were treated at a specialized COVID-19 isolation ward built at the Dow University of Health Sciences at the Ojha campus. The patients were referred from the emergency department, medical and allied wards, and COVID-19 screening units. A detailed history and clinical examination were performed, and comorbidities were evaluated. Results A total of 212 patients were admitted during the study with a mean age of 52 ± 16 years. The study population consisted of 120 (56.6%) males and 92 (43.39%) females, and the most common comorbidities were uncontrolled diabetes with hypertension (n = 56; 26.4%), controlled diabetes (n = 22; 10.37%), obstructive airway disease (n = 16; 7.5%), and interstitial lung disease (n = 14; 6.6%). A total of 48 (22.64%) patients had no comorbidities. Conclusions Most COVID-19-positive patients with pneumonia were male, and common comorbidities included uncontrolled diabetes, hypertension, and obstructive and restrictive lung disease. The presence of comorbidities was associated with a marked increase in the risk of morbidity and mortality. Further studies are warranted to confirm these findings.

Neonatal calf diarrhea: A potent reservoir of multi-drug resistant bacteria, environmental contamination and public health hazard in Pakistan.

Though emergence of multi-drug resistant bacteria in the environment is a demonstrated worldwide phenomenon, limited research is reported about the prevalence of resistant bacteria in fecal ecology of neonatal calf diarrhea (NCD) animals in Pakistan. The present study aimed to identify and assess the prevalence of bacterial pathogens and their resistance potential in the fecal ecology of NCD diseased animals of Pakistan. The presence of antibiotic resistance genes (blaTEM, blaNDM-1, blaCTX-M, qnrS) was also investigated. A total of 51 bacterial isolates were recovered from feces of young diarrheic animals (n = 11), collected from 7 cities of Pakistan and identified on the basis of 16S rRNA gene sequence and phylogenetic analysis. Selected isolates were subjected to antimicrobial susceptibility by disc diffusion method while polymerase chain reaction (PCR) was used to characterize the blaTEM, blaNDM-1, blaCTX-M, qnrS and mcr-1 antibiotic resistance genes. Based on the 16S rRNA gene sequences (Accession numbers: LC488898 to LC488948), all isolates were identified that belonged to seventeen genera with the highest prevalence rate for phylum Proteobacteria and genus Bacillus (23%). Antibiotic susceptibility explained the prevalence of resistance in isolates ciprofloxacin (100%), ampicillin (100%), sulfamethoxazole-trimethoprim (85%), tetracycline (75%), amoxicillin (55%), ofloxacin (50%), ceftazidime (45%), amoxicillin/clavulanic acid (45%), levofloxacin (30%), cefpodoxime (25%), cefotaxime (25%), cefotaxime/clavulanic acid (20%), and imipenem (10%). MICs demonstrated that almost 90% isolates were multi-drug resistant (against at least three antibiotics), specially against ciprofloxacin, and tetracycline with the highest resistance levels for Shigella sp. (NCCP-421) (MIC-CIP up to 75 µg mL-1) and Escherichia sp. (NCCP-432) (MIC-TET up to 250 µg mL-1). PCR-assisted detection of antibiotic resistance genes showed that 54% isolates were positive for blaTEM gene, 7% isolates were positive for blaCTX-M gene, 23% isolates were positive for each of qnrS and mcr-1 genes, 23% isolates were co-positive in combinations of qnrS and mcr-1 genes and blaTEM and mcr-1 genes, whereas none of the isolate showed presence of blaNDM-1 gene.

Draft genome sequence of a multidrug-resistant novel candidate Pseudomonas sp. NCCP-436T isolated from faeces of a bovine host in Pakistan.

OBJECTIVES: Here we describe the first draft genome analysis of a CRISPR-carrying, multidrug-resistant, candidate novel Pseudomonas sp. NCCP-436T isolated from faeces of a neonatal diarrhoeic calf. METHODS: The genome of strain NCCP-436T was sequenced using an Illumina NovaSeq PE150 platform and analysed using various bioinformatic tools. The virulence factors and resistome were identified using PATRIC and CARD servers, while CGView Server was used to construct a circular genome map. Antimicrobial susceptibility was determined by the disk diffusion technique. RESULTS: The draft genome of strain NCCP-436T contains 43 contigs with a total genome size of 3,683,517 bp (61.4% GC content). There are 3,452 predicted genes, including 60 tRNAs, 7 rRNAs and 12 sRNAs. CRISPR analysis revealed two CRISPR arrays with lengths of 1103 bp and 867 bp. Strain NCCP-436T was highly resistant to fluoroquinolone, ß-lactam, cephalosporin, aminoglycoside, penicillin, rifamycin, macrolide, glycopeptide, trimethoprim/sulfonamide and tetracycline antibiotic classes. Additionally, 22 antibiotic resistance genes, 313 virulence genes and 253 pathogen-host interactor genes were predicted. Comparison of the average nucleotide identity and digital DNA-DNA hybridisation values with the closely-related strain Pseudomonas khazarica (TBZ2) was found to be 82.08% and 34.90%, respectively, illustrating strain NCCP-436T as a potentially new species of Pseudomonas. CONCLUSION: Substantial number of antibiotic resistance and virulence genes and homology with human pathogens were predicted, exposing the pathogenic and zoonotic potential of strain NCCP-436T to public health. These findings may be used to better understand the genomic epidemiological features and drug resistance mechanisms of pathogenic Pseudomonas spp. in Pakistan.

A Rare Case of Hyperglycemic-Hemichorea in a Young Patient.

Chorea is an abnormal, nonrhythmic, and purposeless movement of limbs. There is a long list of diseases responsible for chorea; long-standing hyperglycemia can sometimes result in it, which typically manifests on one side of the body. MRI brain is an added diagnostic tool, which commonly shows hyperintense basal ganglia lesion on T1-weighted images. Chorea in the context of hyperglycemia is a reversible and infrequent occurrence, best managed with insulin and haloperidol combination therapy. Here, we discuss a patient with hyperglycemic-hemichorea, whose symptoms resolved completely within two months of taking insulin and haloperidol.

Frequency of Neurological Presentations of Coronavirus Disease in Patients Presenting to a Tertiary Care Hospital During the 2019 Coronavirus Disease Pandemic.

Background Coronavirus disease 2019 (COVID-19), caused by infection with the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), usually presents clinically with cough, fever, shortness of breath, and loss of taste and/or smell. COVID-19 can also present with neurologic signs and symptoms, including headache, hyposmia/anosmia, encephalopathy, meningoencephalitis, Guillain-Barré syndrome, stroke, and seizure. Viral transmission occurs through aerosols generated when an infected person coughs, sneezes, or exhales and by direct touching of contaminated surfaces. The present study evaluated the frequency of neurologic presentations of coronavirus disease in patients presenting at a tertiary care hospital during the COVID-19 pandemic. Methodology This cross-sectional study included 350 inpatients and outpatients (self-isolated) with polymerase chain reaction-confirmed SARS-CoV-2 infection who presented at Dow International Medical College of Karachi between March and June 2020. Of these 350 patients, 68 (18.9%) presented with neurological signs and symptoms and were further evaluated. The data were analyzed statistically using IBM Statistical Product and Service Solutions (SPSS) for Windows, version 20.0 (IBM Corp., Armonk, NY). Results The 350 patients with SARS-CoV-2 infection included 245 (70%) men and 105 (30%) women; of these, 262 (74.9%) were married, and 88 (25.1%) were unmarried. Patients ranged in age from 17 to 88 years (mean ± standard deviation, 49.5 ± 17.4 years), with 68 (18.9%) having neurological manifestations. Headache was the most frequent neurological symptom, reported in 21 (6%) patients, followed by vertigo in 12 patients (3.4%), numbness/paresthesia in 11 (3.1%), altered level of consciousness in seven (2%), hyposmia/anosmia in five (1.4%), and encephalitis in three (0.9%). Other symptoms included sudden hemiparesis (stroke) in two patients (0.6%), flaccid paralysis due to Guillain-Barre syndrome in one (0.3%), and seizure in one (0.3%). Conclusion Neurological involvement is not infrequent in patients with COVID-19. Neurologic manifestations should be carefully monitored in infected patients. COVID-19 should be suspected in patients presenting with neurological abnormalities and should be included in the differential diagnosis to prevent further virus transmission.

Clinical Features Indicating the Need for Mechanical Ventilation in Patients with Guillain Barre Syndrome.

Background Guillain Barre Syndrome (GBS) is an immune-mediated inflammatory polyradiculoneuropathy. Respiratory failure is one of its recognized and most dreaded complications, requiring ventilatory assistance. Early recognition of distinct clinical predictors of mechanical ventilation may help in the better management of GBS patients in our setup. Objective To determine the clinical predictors indicating the need for mechanical ventilation in patients with Guillain Barre Syndrome and to compare the presenting features in patients who require mechanical ventilation and who do not. Method It was a prospective observational study. A total of 100 consecutive patients, over the period of one year, were included in this study. All patients were clinically examined for limb weakness, neck weakness, bulbar and facial nerve involvement, and followed up till seven days of hospitalization for whether the patient required mechanical ventilation or not. Results were recorded on a specifically designed proforma. Data were entered and analyzed using SPSS version 20.0 (IBM Corp., Armonk, NY, US). Results Out of 100 patients, 13% required mechanical ventilation. When clinical presentations were compared in patients who required mechanical ventilation and those who did not, a shorter interval between the onset of symptoms and the attainment of maximal disability, facial weakness, bulbar dysfunction, and neck weakness turned out to be significant factors (p-value<0.000). Conclusion According to these significant outcomes of our study, the course of patients with GBS leading to mechanical ventilation can be predicted on the basis of clinical presentations. So we can recommend that shorter time duration between symptom onset and peak disability, along with the presence of facial, bulbar, and neck weakness, should be taken as an indication of impending respiratory failure.