International Journal of Applied and Experimental Biology

Appraisal of phytotoxicity of heavy metals in radish (Raphanus sativus L.) grown in Sargodha district, Pakistan: A quantitative assessment

Zafar Iqbal Khan — 2026-01-01

The freshwater shortage and increased domestic effluents have led the farmers to utilize untreated sewage water for irrigation, owing to population growth and urbanization in Pakistan. Though sewage water contains essential plant nutrients, it is also a foremost source of metal contamination within the food chain. This study aimed to compare heavy metals (Cd, Fe, Mn, Cu, Zn, Pb, Ni, and Cr) contamination in radish (Raphanus sativus L.) grown in soils irrigated with sewage water, tube well water, and canal water in Sillanwali and Sargodha of Punjab, Pakistan. The areas were evaluated for heavy metal contamination using soil quality indices, including contamination factor (CF), enrichment factor (EF), bioconcentration factor (BCF), estimated daily intake (EDI), and hazard quotient (HQ), which were duly computed for the collected samples. The physicochemical properties of the soil and water samples were also analyzed. The results indicated that the sewage-irrigated areas are more vulnerable to risk concerning metal contamination than those located along canals. Metal concentrations in sewage-irrigated areas were significantly higher in the edible parts of the vegetables; sewage water contained and transferred more metals than canal water; hence, it poses increased health and ecological risks. Peak concentrations were observed at site SW1 (sewage water, site 1) and were the lowest at site TW2 (tube well water irrigation, site 2). Since all metal index values fell below 1, it shows that all metal concentrations were within permissible limits.

Adaptive traits in plants for salinity stress tolerance: A critical review

Fahad Shafiq — 2026-01-01

Soil salinity negatively affects plants at multiple levels, inducing ionic and metabolic imbalances that directly affect growth and productivity. Plants counter salinity stress through a combination of adaptive traits that facilitate cellular ion homeostasis and prevent excessive sodium (Na⁺) accumulation or its detrimental effects. This review provides in-depth information focusing on plant salinity tolerance mechanisms. The Na⁺-exclusion, xylem loading, and Na⁺-vacuolar sequestration by compartmentalization are primarily achieved by the set of antiporters (HKT1, NHX1, NHX2, and SOS1) located at the root cell plasma membrane, xylem parenchyma, and the tonoplast. Plants regulate Na⁺ xylem loading to restrict Na⁺ translocation to aerial tissues, mediated by SOS1 and HKT1. Cellular compartmentalization is regulated by both Na⁺/H⁺ and K⁺/H⁺ antiporters, which maintain ion homeostasis. Osmoregulation in cells is achieved with the help of both organic osmolytes (proline, glycine betaine, sugars, polyols, etc.) and inorganic ions (Na⁺, K⁺, Ca²⁺), which counter salt-induced osmotic stress. However, oxidative stress is mitigated by various enzymatic antioxidant proteins (SOD, CAT, POD, APX, GR, DHAR) and non-enzymatic antioxidant molecules (e.g., vitamin C, glutathione, etc.). Apart from that, the roles of various hormones, for instance, abscisic acid, ethylene, salicylic acid, jasmonates, brassinosteroids, auxins, gibberellins, and cytokinins are discussed at length in this review.

Floral diversity and ethnobotanical profile of Anwar-Mahal, District Charsadda, Khyber Pakhtunkhwa, Pakistan

Fazal Hadi — 2026-01-01

To organize floral diversity and explore the ethnobotanical knowledge of Anwar-Mahal Village, District Charsadda, Pakistan, the plants were collected and documented. The indigenous knowledge about plant use was obtained from the local community, and selected plant species were analyzed for their nutritional profile. Plants were collected during field visits to the study area and were identified with the help of available literature. For ethnobotanical data collection, the questionnaire method was adopted, and Informants were randomly selected from different age groups. The artifact method (ex-situ sampling) and the inventory method (in-situ sampling) were used for the same. Elemental analysis of the selected species was performed at the Centralized Resource Laboratory, University of Peshawar, Pakistan, and proximate analysis was done at the Nuclear Institute for Food and Agriculture (NIFA), Peshawar, including moisture content, ash analysis, fibers, carbohydrates, and protein. The floral diversity of the area consists of 166 species distributed in 120 genera and 47 families. Four families were monocots with 9 genera and 23 (13.86%) species, and 43 families were dicots with 111 genera and 143 (86.14%) species. Poaceae was the leading family with 17 (10.24%) species, followed by Asteraceae with 16 (9.63%) species. The number of herbs was 119 (71.69%), trees 26 (15.66%), and shrubs 21 (12.65%) species. Therophyte was the dominant life form, having 84 (50.29%) species, and microphyll with 62 (36.68%) species was the leading leaf size class. Of different species recorded, 75 (34%) were medicinal, 35 (16%) fodder, 30 (14%) fuel, 20 (09%) vegetable, 17 (08%) fruit, 13 (06%) timber, 12 (05%) ornamental, 9 (04%) condiments, 7 (03%) beverages, and 3 (1.4%) poisonous. Elements like Mg, Ca, K, Na, Cu, Fe, Mn, and Zn were found to be in varying quantities in the selected plant species.

Eco-physiological adaptive strategies of Prosopis cineraria (L.) Druce in response to the diverse edaphic habitats of the Cholistan Desert, Pakistan

Muhammad Akram — 2026-01-01

Salt stress is a significant environmental issue that harmfully affects plant growth and metabolism. However, plants tend to adapt to salt stress by regulating their biochemical and physiological attributes. Physio-biochemical responses to stress conditions of natural habitats remain unclear in Prosopis cineraria. This study evaluated the eco-physiological and biochemical adaptive strategies in P. cineraria growing under different edaphic habitats of the Cholistan Desert. Three edaphic habitats (Sand dune = SD, sandy plain = SP, saline area = SA) of the Cholistan desert were selected, each with three sub-sites as replicates. For the ecological study, five randomly positioned 10 m × 10 m quadrats were taken at each selected site. Physicochemical results revealed that at SD, EC was 9.4 dS m⁻¹, pH 7.5, and organic matter (OM) 0.78%. At SP, EC increased to 157.7%, pH to 9.33%, and OM to 20%. The soil Na⁺ level at SD was 1650.8 mg L⁻¹, but at SA, it was 200.8% compared to that of SD. The physiological attributes of the plant were significantly modified in a high-saline area, as chlorophyll a and b declined considerably by 34.26% and 54.32%, respectively, at SA compared to those of SD. Total soluble proteins, free amino acids, soluble sugars, proline, and root tissue contents of Na⁺, K⁺, Ca²⁺, and Cl^- were considerably higher at SA. In wild populations, these physiological changes appeared to be crucial for sustainable survival in the arid, saline desert environment. Prosopis cineraria was identified in all edaphic habitats and showed ecological dominance at the saline area in the Cholistan Desert, indicating its particular adaptability to the salty environment.

Mitigating chromium toxicity in oat (Avena sativa L.) with exogenous salicylic acid

Hafiz Muhammad Mohsin Hassan — 2026-01-01

This study investigated the protective role of exogenous salicylic acid (SA) in mitigating chromium (Cr) toxicity in oat (Avena sativa L.). A pot experiment was conducted in a shade house using a factorial design with six treatment groups: Control (No Cr, No SA), CrCl₃ 2 mM, SA 0.5 mM, SA 2 mM, CrCl₃2 mM + SA 0.5 mM, CrCl₃2 mM + SA 2.0 mM. Chromium stress significantly reduced plant growth, yield, and anatomical parameters. Chromium treatment significantly decreased the root-shoot ratio, while both concentrations of SA improved growth considerably. The SA (2.0 mM) treatment proved to be most effective in enhancing overall plant growth and grain yield of oat. Anatomical analysis revealed that SA application mitigated the damage induced by Cr. Specifically, the thickness and area of stem sclerenchyma were significantly increased by 1.3- and 1.5-fold, respectively, with the 2.0 mM SA treatment compared to those in the control. Furthermore, SA improved the thickness of the leaf midrib, metaxylem, and phloem, as well as the root epidermis, endodermis, and phloem, particularly in the Cr + SA treatment. These anatomical changes are crucial for water conservation and overall plant survival under heavy metal stress. Our findings suggest that exogenous application of SA can effectively alleviate Cr-induced stress, promoting the healthy development of oat seedlings.

Structural and functional traits mediating the adaptability of creeping signal grass [Urochloa reptans (L.) Stapf] across diverse ecozones

Hafeez ur Rehman — 2026-01-01

Plants of the genus Urochloa P. Beauv., popularly known as brachiaria, or marmalade grass, belong to the family Poaceae. Urochloa reptans is a member of the genus Urochloa and is also known as creeping signal grass. It is widely distributed in tropical and subtropical regions of the world. The present experiment explored the anatomical, structural, functional, and physiological adaptive features of Urochloa reptans sampled from sixteen different sites of the Faisalabad region, Pakistan. All the collected populations showed numerous anatomical modifications, and also possess marked physiological characteristics that help them to survive under varying ecological conditions. This leads to their ecological success for existence in heterogeneous environments. The populations of Eden Garden and Canal Road had maximum leaf epidermal thickness, while midrib thickness was higher in the Manawala ecotype. The bulliform cell area was noted to be widened mainly in the Narwala Bangla population. The intensive sclerification of the stem was observed in a sample of Makkuana. The populations from Painsra and Eden Garden possessed larger vascular bundle areas. However, only the Eden Garden population exhibited a larger metaxylem area. The populations native to Canal Road and Sadar Bypass exhibited vigorous root growth in terms of cortical region thickness, endodermal thickness, and pith cell area. Besides this, well-developed aerenchyma cells and metaxylem vessels were observed in the Canal Road and Chicha populations. Overall, U. reptans showed much outstanding activity in the context of development, structural, functional, and physiological characteristics in response to the present-day ecological scenario.

Critical period for alligator weed (Alternanthera philoxeroides) in puddled transplanted rice

Azhar Mehmood — 2026-01-01

Alligator weed (Alternanthera philoxeroides), an invasive plant in Pakistan due to its amphibious growth habit, has become a problematic weed for puddled transplanted rice (PTR). To assess the critical timing of removal (CTWR) and rice yield losses caused by this weed, a two-year field investigation was conducted. The treatments included varying alligator weed-crop competition durations for the initial 4 weeks following transplanting (WFT), 6 WFT, 8 WFT, and 10 WFT. The whole season, weedy and weed-free treatments were also kept. During both years, a gradual decline in paddy yield and rice quality parameters occurred with the prolongation in weedy period due to an increase in alligator weed’s dry weight (up to 196%), N-uptake (up to 114%), P-uptake (up to124%), K-uptake (up to 134%), Cu-uptake (up to 101%), Fe-uptake (up to 21%), Zn-uptake (up to 81%), and Mn-uptake (up to 58%). Consequently, the full-season alligator weed competition resulted in the highest reductions in paddy yield (21.6%-22.3%), fertile tillers per hill (10.5%-11.5%), kernels per panicle (3.1%-3.7%), 1000-kernel weight (15.4%-21.2%), kernel amylose content (12.9%-13.5%), and protein content (12.5%-21.6%) during the first and second years’ experiments, respectively. The alligator weed infestation increased in percentages of sterile spikelets (56%-69%), opaque kernels (27%-28%), and chalky kernels (17%-18%) of rice. The logistic model estimated the CTWR of the alligator weed in puddled transplanted rice (PTR) to be 6.1 WFT and 5.7 WFT in order to prevent 5% yield losses during the years 2011 and 2012.

Adaptations and mechanisms underlying plant resilience to drought: Opportunities, challenges, and prospects for water-saving agriculture

Asifa Batool — 2026-01-01

One of the major obstacles to plant growth and productivity, outside of today's crop-growing zones, is drought. Due to acknowledged changes in the global climate, it will become more significant in areas of the world where the issue was previously insignificant. Considering the mechanisms underlying resistance to water deficit and the plants' effective water use is essential to addressing current concerns about enhancing plant genotypes and cultural practices for drought-affected locations. Main barriers to carbon uptake and the metabolic processes that influence how plants react to water scarcity, either alone or in combination with other stressors, are covered in this work. For crops that suffer from drought, oxidative stress is essential. Detoxifying mechanisms play a key role in preventing irreparable damage to redox molecules and photosynthetic machinery as a result of local or systemic signals. The ability of plants, particularly those that embed functional proteins, to prevent or fix membrane damage that occurs throughout the processes of dehydration and rehydration is essential to maintain the integrity of the membranes. These proteins include water transporters, whose function in controlling the water status of plants and moving other metabolites is being thoroughly studied. Over ten years ago, long-distance chemical signaling began to unravel as a primary response to water deficit. In light of new management strategies, the distribution and absorption of carbon and its assimilates among non-reproductive and reproductive parts are reviewed and examined. Enhancement of water-use efficiency in plants, consistent production, and better quality in products are all intended outcomes of these applications. It has already been feasible to discover important genes that can change metabolic processes, and boost the tolerance of plants to drying conditions by studying mechanisms underlying successful response to dehydration and rehydration. This study provides a summary of the most significant information on this subject, including water transporters and engineering for C₄ characteristics. Promising technologies, including the modification of crops by genetic engineering via the transfer of regulatory and functional genes, are highlighted.

Clustering of date palm (Phoenix dactylifera L.) cultivars using morphological and yield attributes as selection criteria

Mehwish Naseer — 2026-01-01

Thirty-two (32) date palm (Phoenix dactylifera L.) cultivars of diverse origin were characterized morphologically to assess the extent of inter-cultivar variability. These cultivars originated in parts of Pakistan, Iran, Iraq, and Saudi Arabia. A great magnitude of inter-cultivar variation was found in the set of date palm cultivars, not only in quantitative, but also in qualitative morphological characteristics. A multivariate (cluster) analysis of the date palm cultivars showed a lesser degree of similarity in qualitative morphological characters as compared to quantitative morphological characters. Based on qualitative morphological characters of the trunk and leaves, most of the cultivars from Punjab and two from Baluchistan were closely similar to each other, and a similar pattern of similarity was noted in their quantitative morphological characteristics. Characteristics like color of spines, number of leaflets per leaf, and size, shape, and color of fruits and seeds were found to be suitable morphological selection criteria for the exact identification of date palm cultivars. Date palm cultivars Dakki (from Khyber Pakhtunkhwa) and Chawara (from Baluchistan), and Hallawi-I (from Iraq) ranked better regarding fruit size, weight, and taste quality. Khudrawi (from Iraq) and Saib (from Punjab) were also likable, having large and tasty fruits.

Allicin improves gas exchange characteristics, redox balance, and ion homeostasis to mitigate chromium toxicity in wheat

Umer Farooq — 2026-01-01

This research explored how exogenous application of allicin can assist wheat plants in coping with the stress caused by chromium (Cr). The findings indicate that Cr toxicity (30 mg kg^-1) conspicuously reduced biomass, chlorophyll content, and leaf relative water content (LRWC) in wheat plants. A significant surge in oxidative stress, as demonstrated by higher levels of hydrogen peroxide, superoxide radicals, and malondialdehyde alongside elevated lipoxygenase activity and electrolyte leakage percentage, was noticed in Cr-stressed plants. Chromium toxicity also induced higher methylglyoxal generation in plants. Allicin foliar administration (50, 100, and 200 µM) visibly curtailed Cr phytotoxicity and improved biomass. Allicin applied as 200 µM maximally subsided oxidative stress by augmenting the activities of superoxide dismutase (SOD), peroxidase (POD), catalase(CAT), and ascorbate peroxidase (APX), as well as the levels of ascorbate and reduced glutathione (GSH). Higher endogenous hydrogen sulfide (H₂S) and nitric oxide (NO) concentrations in allicin-treated plants further stimulated the antioxidant defense system of Cr-challenged plants. Allicin supplementation maintained GSH : GSSG ratio in Cr-stressed plants, suggesting a better redox balance. A noticeable accretion of soluble sugars, flavonoids, and phenolics due to allicin safeguarded plants from metal-induced damage and its associated oxidative stress. These findings highlight that exogenous application of allicin can be used as a promising strategy to alleviate Cr toxicity in wheat plants. Future studies should assess the effectiveness of exogenous allicin application in other cereal crops under controlled and field conditions. Additionally, exploring the molecular mechanisms underlying metal stress tolerance in allicin-treated plants will deepen our understanding.

Assessment of phosphate-solubilizing rhizobacterial strains combined with rock phosphate and compost and their effect on the growth and yield of maize (Zea mays L.) under field conditions

Azhar Razaq — 2026-01-01

Adequate phosphorus (P) supply is critical for optimum plant growth. However, soils enriched in total P content frequently do not fulfill the plant requirements due to the provided P, which may become unavailable in alkaline calcareous soils. The use of slow-release P fertilizers, such as rock phosphate (RP), compost, and plant growth-promoting rhizobacteria (PGPR) with P-solubilizing activity, can stimulate plant growth by solubilizing inaccessible P in the soil. A field study was conducted to estimate the efficacy of PGPR containing P-solubilizing and ACC-deaminase activity in soil amended with RP and compost on the production and growth of maize. Ten pre-isolated bacterial isolates were taken and tested for their effect on maize with individual and combined application of PSB, RP, and compost with a control. Overall, bacterial isolates (S5 and S6) containing P-solubilizing and ACC-deaminase activity combined with RP and compost significantly improved the maize crop development and production over the control. Finally, it is suggested that RP could be efficiently applied combined with compost and P-solubilizers.

Changes in gut microbiota, hematological parameters, and immune response following dietary administration of oyster mushroom (Pleurotus ostreatus) in silver carp (Hypophthalmichthys molitrix)

Muhammad Haroon — 2026-01-01

The primary aim was to evaluate the impact of oyster mushroom (Pleurotus ostreatus) on the gut microbiota, hematological parameters, and immune response of silver carp (Hypophthalmichthys molitrix). The experimental group was given P. ostreatus powder at 2% of body weight, while the control group was fed with commercial fish feed. Fish were treated with these two feeds for 8 weeks. Culturing of bacteria, hematological tests, and immunological assays were done to evaluate the effect of these feeds on H. molitrix. Physico-chemical parameters of the aquarium were maintained end-to-end during the trial period, such as DO at 5-7 mg L^-1, temperature at 24-28 °C, and pH at 7.0-8.2. The quantitative outcomes from the groups were compared using the t-test. In all three culturing media, such as nutrient agar (NA), tryptic soy agar (TSA), and eosin methylene blue (EMB), the results indicated a significant decrease in bacterial colonies in the treatment group, when compared with the control group. Compared to the control group, the treatment group had a significantly higher RBC count, higher hemoglobin value, enhanced hematocrit levels, MCV, and MCHC. White blood cells (WBCs), neutrophils, and lymphocytes in the treatment group had a significantly higher count than those of the control group. However, the hematological parameter MCH and immune cells, such as monocytes, remained unchanged in the treatment and control groups. Hence, using immunostimulants such as mushrooms in aquaculture increases the body’s natural resistance to infection and facilitates the prevention of various diseases.

Impact of banana peel and cocoa husk on growth performance, microbial communities, and skin pigmentation in goldfish (Carassius auratus) and Thaila (Catla catla)

Aqsa Ameen — 2026-01-01

Carassius auratus and Catla catla are polyculture species where growth and intensification have adverse effects on water quality, raising the risk of infection and disease prevalence. Due to these factors, the survival rate of farm fish is reduced. The demand for aquaculture continues to rise with human population growth, emphasizing the importance of well-formulated feeds in aquaculture success. High-quality feed can be prepared using industrial byproducts. It leads to minimizing feed expense, controlling disease outbreaks, and increasing net profit. This research work was conducted to study the impact of banana peel and cocoa husk on growth, microbial community, and pigmentation in C. catla and C. auratus. For this purpose, 30 specimens each of C. catla and C. auratus were randomly assigned to three groups: control (T₀) and experimental (T₁ and T₂). T₀ received commercial feed, whereas T₁ and T₂ were fed banana peel and cocoa husk diets at 2% body weight, respectively. For statistical analysis, a 2-factor factorial under CRD was performed. In the experimental group, length, weight, and specific growth rate (SGR) were non-significant between the species, but showed significant differences compared to the control group. In comparison to the control group, the bacterial count and pigmentation in the experimental group were significant. Because of their carotenoids and antioxidant qualities, banana peel and cocoa husk increased both species' immunological responses and pigmentation. Finally, integrating banana peel and cocoa husk into the diet of C. auratus and C. catla holds potential benefits in terms of reducing intestinal microorganisms, improving growth, and enhancing pigmentation of the fish species.

Effect of soybean meal (SBM) on growth and hematological parameters of rohu (Labeo rohita) and grass carp (Ctenopharyngodon idella)

Areeha Fatima — 2026-01-01

This study investigates the efficacy of soybean meal (SBM) as a potential protein source in aquafeed, focusing on its effects on the growth and hematological parameters of rohu (Labeo rohita) and grass carp (Ctenopharyngodon idella). The research was conducted at the Microbiology and Immunology Laboratory, University of Agriculture, Faisalabad. With global demand for fishmeal exceeding supply, alternative protein sources, such as SBM, are increasingly crucial for sustainable aquaculture. The study, spanning 7 weeks, involved assigning rohu and grass carp into control (fish meal) and experimental soybean meal (SBM) groups. Growth parameters, including weight, length, specific growth rate (SGR), feed conversion ratio (FCR), and survival rate, were monitored, alongside hematological analyses. The fish were fed on an experimental diet at a feeding level of 2% of body weight. The results indicated comparable growth performance between SBM-fed and fishmeal-fed groups, with SBM potentially enhancing survival rates. Body weight gain and SGR increased significantly compared to the control group fed with FM in rohu. However, for grass carp, there was no significant difference in weight gain and SGR values when fed SBM compared to FM. For rohu, the hematological studies showed significantly higher red blood cells, hemoglobin, hematocrit, white blood cells, and leucocytes. For grass carp RBCs, hemoglobin, and hematocrit values showed a declining tendency with experimental feed; besides that, there was a prominent rise in white blood cell (WBC) count and lymphocytes. These results suggested that SBM could be incorporated at varying percentages in carp feed, considering enhanced growth performance and hematological status as a cost-effective alternative protein source strategy to overcome the cost of fish meal protein in the aquaculture industry.

Effect of carrot (Daucus carota) flour on pigmentation and blood parameters of Catla catla and Cyprinus carpio

Ifrah Mustafa — 2026-01-01

The primary aim of this study was to determine the effect of carrot flour (Daucus carota) on pigmentation and blood parameters of Catla catla (Thaila) and Cyprinus carpio (Common Carp). Pigmentation is often a key indicator for fish in terms of consumer acceptability. Thaila has immense cultural and economic importance in South Asia because of its flesh quality and large size. Common carp is also an important edible fish and the third most frequently introduced species all over the world due to its ornamental colors. Carrot flour, rich in vitamin A, is a natural vegetable feed source as a pigmentation enhancer because it contains carotenoids in large amounts. High amounts of beta-carotene also act as an antioxidant and have a beneficial effect on fish health. For this experiment, an 8-week trial was conducted with two groups (one experimental and one control) in triplicate, having an equal number of fish (n = 20 each). After completion of the trial, blood and skin samples from both fish species were taken and subjected to a hemocytometer and a spectrophotometer, respectively. Statistical inference using the t-test was done to get the final results of the trial. The carrot flour supplementation led to increased red blood cell counts in both species, with significant changes in other hematological parameters as well. Additionally, both species exhibited enhanced pigmentation in dorsal and ventral scales when fed carrot flour according to 2.5% of the body weight of the fish. These findings suggest potential benefits of the carrot flour in improving hematological parameters as well as pigmentation in fish, leading to better consumer acceptability and potential economic benefits.

Cytoprotective role of p-cymene against alcohol-induced toxicity in human embryonic kidney cells

Shabana Akhtar — 2026-01-01

p-Cymene (p-isopropyl toluene) is a monocyclic monoterpene and is the main compound of many aromatic plant species. p-cymene also exhibits various pharmacological properties, including antimicrobial, antioxidant, anti-inflammatory, antidiabetic, antiviral, and antitumor activities. p-cymene also acts as a strong anti-inflammatory and antioxidant agent. Alcohol use is widespread in the Western world and is found to be nephrotoxic, and can cause kidney injury, especially during pregnancy and kidney development during intrauterine life. Alcohol can lead to reduced levels of antioxidants and increased synthesis of pro-inflammatory markers. This study aimed to investigate the cytoprotective role of p-cymene against alcohol-induced toxicity in human embryonic kidney (HEK293) cells. In this study, we developed four groups: a negative control group, a positive control group, an injury group, and an injury plus treatment group. Injury was induced by ethanol, and then different concentrations of p-cymene, i.e., 50 µM, 100 µM, and 500 µM, were added to check the cytoprotective action of p-cymene. The MTT assay and oxidative stress marker (glutathione reductase) were assessed to check cell viability. The KIM-1 and 8-OHdG ELISA were done to evaluate the apoptotic pathways. The results concluded that the 500 µM treatment increased cell viability more than the other levels. According to the MTT Assay, Crystal Violet Assay, and Trypan Blue Assay, p-cymene increased cell viability in ethanol-injured cells. KIM-1 ELISA showed lower levels of kidney injury molecule-1 in p-cymene-treated cells. While 8-OHdG ELISA showed that 8-hydroxy guanosine levels were also reduced in the cells treated with p-cymene, in contrast to cells left untreated. In the Antioxidant assay, a GSH ELISA was performed, and the glutathione value increased in treated cells. In this research, it has been observed that p-cymene has a cytoprotective role against alcohol-induced toxicity in human embryonic kidney cells.

Insights into molecular characterization and post-vaccination dynamics of Haemophilus influenzae

Maaria Raheel Spaul — 2026-01-01

Hemophilus influenzae is a Gram-negative bacterium exhibiting the characteristics of a facultative anaerobe. The bacterium requires capnophilic conditions for its growth. These bacteria are classified as fastidious organisms because they require X and V factors for their growth. They are present as commensals on the mucosa of the upper respiratory tract of humans. Haemophilus influenzae has no veterinary importance; it is exclusively adapted to its human host. The nontypeable strain (non-encapsulated) is mostly present in the normal flora of humans, but the typeable strain (encapsulated) can also exist as a commensal. To make a precise determination about infectious agents, methods of molecular characterization help distinguish among closely related isolates. Molecular characterization of Haemophilus influenzae is based on biochemical characteristics, capsular types, biotypes, lipopolysaccharide, and lipooligosaccharide, providing valuable insights into pathogenicity, resistance mechanisms, and epidemiology of this exclusive human pathogen. Laboratory diagnosis is done for the precise understanding of vaccine efficacy, disease outbreak monitoring, clinical management, and surveillance purposes. Protein-conjugated vaccine is used for its prevention. This has significantly reduced the incidence of Hib diseases. Still, simultaneously, the emergence of NTHi infections has also been reported, keeping in mind that NTHi is a prominent cause of otitis media, sinusitis, chronic obstructive pulmonary disease (COPD), and other invasive conditions in susceptible populations. However, utilizing surface proteins as a vaccine antigen can pave the way towards protective immunization against the NTHi strains. Additionally, the interdisciplinary relationship of molecular analysis with other areas of genomic research in determining pathogenicity, antibiotic resistance, vaccine efficacy, of pre- and post-vaccinal disease epidemiology is the main subject of this review; therefore, an effective approach towards molecular characterization can pave the way for developing protein-based vaccines for the control of NTHi infections in the post-Hib vaccine era.

Assessment and analysis of polymyxin sensitivity using the disc elution method on clinical isolates resistant to carbapenems

Muhammad Raheel Awan — 2026-01-01

Elevated mortality rates have drawn the focus of researchers to multidrug-resistant Gram-negative Enterobacteriaceae over the last decade. In the case of multidrug-resistant Gram-negative bacteria, particularly carbapenem-resistant Enterobacteriaceae (Enterobacterales), polymyxin B stands as one of the last-resort antibiotics. This study aimed to evaluate clinical isolates of Gram-negative bacteria displaying resistance to carbapenems and subjected them to polymyxin B using the disc diffusion method. Clinical specimens were gathered from various anatomical sites, and bacterial isolates were obtained through culturing. After isolation, biochemical testing was conducted for bacterial identification. The identified isolates were then subjected to antibiotic susceptibility testing, employing the Kirby-Bauer Disc Diffusion technique. A comparative analysis was undertaken between the disc elution method and the disc diffusion method, with validation against the reference Broth Microdilution (BMD) technique. A total of one hundred and fifty-two (152) clinical specimens were obtained. The isolates primarily comprised Klebsiella pneumoniae (n = 49), Escherichia coli (n = 41), Pseudomonas aeruginosa (n = 31), Acinetobacter baumannii (n = 29), and Proteus vulgaris (n = 02). Diverse specimen types were collected from various patient sites, encompassing pus swabs (n = 82), urine (n = 36), tracheal tubes (n = 12), sputum (n = 11), blood cultures (n = 6), bronchial washings (n = 2), high vaginal swabs (n = 2), and cerebrospinal fluid (n = 1). The distribution of patients was categorized by gender, with 95 females and 57 males. The majority of the strains were identified in patients aged between 41 and 60 years. The disc elution method categorized 127 samples as susceptible, 13 as intermediate, and 12 as resistant to polymyxin B, while the disc diffusion method labeled 128 samples as susceptible and 24 as resistant. In routine antimicrobial susceptibility testing, the disc diffusion method is commonly used for polymyxin B, even though it is not recommended by CLSI. This study concludes that the disc elution method, recommended by CLSI, demonstrates superior accuracy, precision, and sensitivity in detecting susceptibility to polymyxin B compared to the disc diffusion method.