Comparative genomics and evolutionary analysis of dengue virus strains circulating in Pakistan.

Dengue fever virus (DENV) poses a significant public health risk in tropical and subtropical regions across the world. Although the dengue fever virus (DENV) exhibits significant genetic diversity and has the potential to evolve, there is a lack of comprehensive research on the comparative genomics and evolutionary dynamics of the virus in Pakistan. Phylogenetic analysis demonstrated the circulation of all four dengue virus serotypes (DENV-1, - 2, - 3, and - 4) with prevalent genotypes III and V within DENV-1, cosmopolitan genotype within DENV-2, genotype III within DENV-3, and genotype I within DENV-4 during 2006-2014. Based on the complete envelope region, genome-wide residue signature and genetic diversity indicate that there is a high level of genetic diversity among DENV-1 strains, while DENV-3 strains exhibit the least genetic diversity. Comparative analysis of all four DENV serotypes revealed that certain codons in DENV-2 and -4 were subject to strong purifying selection, while a few codon sites in the envelope region showed evidence of positive selection. These findings provided valuable insights into the comparative genomics and evolutionary pattern of DENV strains reported from Pakistan. Whether those characteristics conferred a fitness advantage to DENV-1 genotypes within a specific geography and time interval warrants further investigations. The findings of the current study will contribute to tracking disease dynamics, understanding virus transmission and evolution, and formulating effective disease control strategies.

Multi-functional application of octacosanol as a feed additive in animal and aquaculture: A review.

Demand for sustainable animal and aquaculture production drives the exploration of novel feed additives. We highlight octacosanol, a long-chain alcohol from plant sources, as a promising multifunctional feed additive. The review comprehensively evaluates octacosanol's applications in animal and aquaculture nutrition, including its molecular properties and mechanisms of action. It elucidates how octacosanol affects lipid metabolism, energy utilization and immune modulation. Octacosanol enhances livestock growth, efficiency, carcass quality and stress resilience. We thoroughly discuss how it enhances feed utilization, disease resistance and overall performance in finfish and shellfish in aquaculture. The review also addresses the ecological and sustainability aspects of octacosanol utilization. We identify challenges and knowledge gaps in octacosanol research, prompting suggestions for future investigations. We address regulatory considerations, dosage optimization and potential interactions with other feed additives to ensure the safe and effective use of octacosanol. In conclusion, the review highlights octacosanol's potential as a versatile feed additive in the animal and aquaculture industries and urges further research to uncover its benefits and sustainability contributions, proposing a prospective research plan for this purpose. This thorough analysis is a valuable resource for researchers, nutritionists and industry professionals looking to find innovative methods to improve production practices and advance sustainable food systems.

Exploring the Interplay of the CRISPR-CAS System with Antibiotic Resistance in Staphylococcus aureus: A Poultry Meat Study from Lahore, Pakistan.

Staphylococcus aureus is one of the major pathogens responsible for causing food poisoning worldwide. The emergence of antibiotic resistance in this bacterium is influenced by various factors. Among them, bacterial acquired defense systems described as clustered regularly interspaced short palindromic repeats (CRISPR)-cas system might be involved in antibiotic resistance development in bacteria. The current study was designed to assess the prevalence of S. aureus and its antibiotic resistance profile and identify the relationship of the CRISPR-cas system with antimicrobial resistance, followed by phylogenetic analysis. Total samples (n = 188) of poultry meat were collected from the poultry bird market of Lahore, Punjab, Pakistan. We used both phenotypic (antibiotic disc diffusion) and genotypic methods (PCR) to identify multi-drug resistant (MDR) strains of S. aureus. Additionally, the role of the CRISPR-Cas system in the isolated MDR S. aureus was also assessed. In addition, real-time quantitative PCR (qRT-PCR) was used to evaluate the association of the CRISPR-cas system with antimicrobial resistance. All of the S. aureus isolates showed 100% resistance against erythromycin, 97.5% were resistant to tetracycline, and 75% were resistant to methicillin. Eleven isolates were MDR in the current study. The CRISPR system was found in all MDR isolates, and fifteen spacers were identified within the CRISPR locus. Furthermore, MDR S. aureus isolates and the standard strain showed higher expression levels of CRISPR-associated genes. The correlation of said system with MDR isolates points to foreign gene acquisition by horizontal transfer. Current knowledge could be utilized to tackle antibiotic-resistant bacteria, mainly S. aureus.

Evaluation of haematological, serum biochemical and oxidative stress parameters in cattle naturally infected with lumpy skin disease virus.

Lumpy skin disease (LSD) is a vector-borne viral transboundary disease of cattle caused by the LSD virus (LSDV). Despite investigations on clinical and outbreak features of LSDV, information on disease pathogenesis and alternative changes in blood parameters are scarce. Keeping this in view, the current study was designed to determine haematological, serum biochemical, and oxidative stress parameters in naturally infected cattle with LSDV during the recent surge of outbreaks in Punjab, Pakistan. A total of 35 blood samples was collected from polymerase chain reaction-confirmed LSDV-infected cattle for assessment of all parameters. The haematological examination of blood samples showed a significant reduction (p < 0.05) in different variables of erythrogram and leucogram. On the other hand, differences between levels of various serum biochemical parameters with the significant increase in levels of alkaline phosphatase, serum alanine aminotransferase, aspartate aminotransferase, and blood urea nitrogen were observed in LSDV naturally infected cattle. Moreover, malondialdehyde levels for lipid peroxidation and nitrate concentration were markedly elevated whereas glutathione S-transferase fluorescent and serum superoxide dismutase enzymes showed a decrease in levels. The current study suggests that alternations in haematological and serum biochemical parameters following LSDV infection stimulate oxidative stress and such findings may be useful for early and rapid diagnosis and improvement in the treatment strategy of the disease.

Levofloxacin induces erythrocyte contraction leading to red cell death.

Background: Levofloxacin, a fluoroquinolone, is an extensively used antibiotic effective against both positively and negatively staining bacteria. It works by inhibiting bacterial topoisomerase type II and topoisomerase type IV, resulting in impaired DNA synthesis and bacterial cell death. Eryptosis is another term for apoptotic cell death of erythrocyte marked by cell shrinkage, phosphatidylserine (PS) flipping, and membrane blebbing. Methods: The intent of the present research was to look at the eryptotic effect of levofloxacin by exposing erythrocytes to therapeutical doses (7, 14 µM) of levofloxacin for 48 hours. Cell size evaluation, PS subjection to outside, and calcium channel inhibition were carried out to investigate eryptosis. Oxidative stress generated by levofloxacin was measured as a putative mechanism of eryptosis using glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase activities. Similarly, hemolysis measurements demonstrated levofloxacin's cytotoxic effect. Results: Our findings showed that therapeutic doses of levofloxacin can cause a considerable decline in antioxidant enzymes activities, as well as induce cell shrinkage, PS externalization, and hemolysis in erythrocytes. The role of calcium in triggering erythrocyte shrinkage was also confirmed. Conclusion: In conclusion, our findings showed that the indicated levofloxacin doses caused oxidative stress, which leads to erythrocyte death via eryptosis and hemolysis. These findings emphasize the importance of using levofloxacin with caution and the need for additional research to mitigate these side effects.

Comprehensive genomic exploration of class III peroxidase genes in guava unravels physiology, evolution, and postharvest storage responses.

Peroxidases (PRXs) play multifaceted roles in plant growth, development, and stress responses. Here, we present a comprehensive analysis of the PRX gene family in guava, a globally significant fruit. In the guava genome, we identified 37 PRX genes, a number lower than that of Arabidopsis, suggesting a distinctive gene family expansion pattern. Phylogenetic analysis unveiled close relationships with Arabidopsis PRXs, with 12 PgPRX genes forming ortholog pairs, indicating a specific expansion pattern. Predictions placed most PRX proteins in the chloroplast and extracellular regions. Structural analysis of PgPRX proteins revealed commonalities in domain structures and motif organization. Synteny analysis underscored the dynamic role of segmental duplication in the evolution of guava's PRX genes. We explored the dynamic expression of PgPRX genes across guava tissues, exposing functional diversity. Furthermore, we examined changes in peroxidase levels and gene expressions during postharvest fruit storage, providing insights for preserving fruit quality. This study offers an initial genome-wide identification and characterization of Class III peroxidases in guava, laying the foundation for future functional analyses.

Xylem anatomical responses of Larix Gmelinii and Pinus Sylvestris influenced by the climate of Daxing'an mountains in Northeastern China.

Wood anatomy and plant hydraulics play a significant role in understanding species-specific responses and their ability to manage rapid environmental changes. This study used the dendro-anatomical approach to assess the anatomical characteristics and their relation to local climate variability in the boreal coniferous tree species Larix gmelinii (Dahurian larch) and Pinus sylvestris var. mongolica (Scots pine) at an altitude range of 660 m to 842 m. We measured the xylem anatomical traits (lumen area (LA), cell wall thickness (CWt), cell counts per ring (CN), ring width (RW), and cell sizes in rings) of both species at four different sites Mangui (MG), Wuerqihan (WEQH), Moredagha (MEDG) and Alihe (ALH) and investigated their relationship with temperature and precipitation of those sites along a latitude gradient. Results showed that all chronologies have strong summer temperature correlations. LA extremes were mostly associated with climatic variation than CWt and RWt. MEDG site species showed an inverse correlation in different growing seasons. The correlation coefficient with temperature indicated significant variations in the May-September months at MG, WEQH, and ALH sites. These results suggest that climatic seasonality changes in the selected sites positively affect hydraulic efficiency (increase in the diameter of the earlywood cells) and the width of the latewood produced in P. sylvestris. In contrast, L. gmelinii showed the opposite response to warm temperatures. It is concluded that xylem anatomical responses of L. gmelinii and P. sylvestris showed varied responses to different climatic factors at different sites. These differences between the two species responses to climate are due to the change of site condition on a large spatial and temporal scale.

Seroprevalence of Newcastle disease virus and avian influenza virus in poultry and captive wild birds in poultry-dense regions of Pakistan.

Newcastle disease virus (NDV) and avian influenza virus (AIV) are causing contagious diseases in chickens and wild birds worldwide; however, there is a paucity of information on the current status of seropositivity of Newcastle and avian influenza diseases in chickens and wild birds of Pakistan. Therefore, the present study aimed to investigate the serological evidence of both diseases in commercial poultry (broiler, layer chickens), backyard poultry, and captive wild birds in poultry­dense regions of Punjab, Pakistan. Enzyme­linked immunosorbent (ELISA) and haemagglutination inhibition (HI) assays were performed for the determination of antibodies against NDV and AIV and their genotyping and subtyping, respectively. Overall, 47.5% and 67.4% seroprevalence of NDV and AIV, respectively, was observed in both poultry and wild birds. Based on bird's category, layer chickens had the highest seroprevalence of NDV (60.8%, 95% CI: 52.95­68.22, OR: 0.71) followed by backyard poultry (56.8%, 95% CI: 47.92­65.32, OR: 0.82), broilers (52.7%, 95% CI: 46.84­58.64), pigeons (41.3%, 95% CI: 30.53­52.81, OR: 1.59), peafowls (26.1%, 95% CI: 11.09­48.69, OR: 3.16), ducks (23.8%, 95% CI: 12.59­39.8, OR: 3.57), turkeys (16.7%, 95% CI: 4.41­42.27, OR: 5.58), parrots (14.3%, 95% CI: 2.52­43.85, OR: 6.70) and quails (2.3%, 95% CI: 0.2­13.51, OR: 4.8). Comparatively, backyard chickens had the highest seroprevalence of AIV (78.8%, 95% CI: 70.64­85.22, OR: 0.63) followed by ducks (73.8%, 95% CI: 57.68­85.6, OR: 0.83), layers (73.5%, 95% CI: 65.98­79.89, OR: 0.84), pigeons (72.5%, 95% CI: 61.2­81.61, OR: 0.89), broilers (70.1%, 95% CI: 64.44­75.29), turkeys (55.5%, 95% CI: 31.35­77.6, OR: 1.87), peafowls (47.8%, 95% CI: 27.42­68.9, OR: 2.56) and parrots (42.8%, 95% CI: 18.8­70.3, OR: 3.1). Overall, 40.1%, 34.2%, 31.3%, and 25.1% sera were positive for H9 AIV, G­VII NDV, H7 AIV, and G­VI NDV, respectively. The current study revealed a widespread exposure to NDV and AIV in poultry and captive wild birds. Therefore, it is crucial to include captive wild birds in NDV and AIV surveillance programs to further strengthen disease control measures, particularly in endemic regions.

Diet impacts on the biological aspects of pink bollworm, Pectinophora gossypiella (Lepidoptera: Gelechiidae) under controlled laboratory conditions.

BACKGROUND: Pink bollworm, Pectinophora gossypiella (Lepidoptera: Gelechiidae) is a native pest of Asia and preferably invasion on cotton (Gossypium hirsutum L.) crop as a commendatory host plant. Commercially, G. hirsutum is known as white gold and is an important cash crop all over the globe. Limited studies were published to focus on certain dietary compositions against different cotton pests. Therefore, the present study was undertaken in the laboratory under controlled conditions (temperature: 27 ± 2°C and relative humidity: 60 ± 10%) to determine the impact of three different treatment diets (wheat germ meal, okra, and chickpea) on the biological aspects (lifetime, developmental period) of P. gossypiella. RESULTS: Results revealed that the shortest larval time of P. gossypiella was observed on the okra feed diet while the longest period was recorded on the wheat germ diet. Meanwhile, the pupation delay was noted on the wheat germ diet. The dietary influence was also observed on adult stages of female and male P. gossypiella (43.00 and 37.50 days respectively) and compared with a standard diet (56.50 and 52.50 days respectively). Furthermore, larval weighed more on the okra and chickpea diet followed by the wheat germ diet, whereas highest pupal weight was observed on the standard diet followed by the chickpea diet and okra diet. CONCLUSION: Developmental parameters were significantly variant across all treatment diets, whereas the higher significant difference was reported on the okra diet. Therefore, the existing data of this study offers fruitful interventions for the future as a modified diet for large-scale and rapid mass production of P. gossypiella larvae.