Multi-drug resistant Acinetobacter baumannii poses a significant threat to public health. This study investigated the genomic features and phenotypic characteristics of two clinical A. baumannii strains, KBN10P01317 (low-virulent) and KBN10P01599 (high-virulent), which share the same sequence type and antimicrobial susceptibility profile. The phenotypic characteristics of A. baumannii strains were assessed by antimicrobial susceptibility testing and virulence trait examination in vitro and in vivo. Whole-genome sequencing was conducted for comparative genomic analysis, and the expression of virulence-associated genes was analyzed using quantitative polymerase chain reaction. Our comparative genomic analysis revealed that KBN10P01599 harbored a larger genome with a greater number of antimicrobial resistance genes, including two copies of the critical resistance gene bla, which might contribute to its higher minimum inhibitory concentration for carbapenems (64 μg/ml) compared to KBN10P01317 (32 μg/ml). Although both A. baumannii strains possessed the same repertoire of virulence-associated genes, KBN10P01599 exhibited significantly enhanced expression of quorum sensing (abaI/R) and biofilm formation genes (csuCDE, bap, and pgaA), correlating with its virulence traits, including increased surface motility, biofilm formation, and adherence to host cells. The differences in the expression of virulence-associated genes between the two strains were partly attributed to the transposition of insertion sequence elements. These findings provide valuable insights into the genetic basis of the virulence potential and antimicrobial resistance in A. baumannii, highlighting the evolutionary changes that may occur within strains of the same clone.

Helicobacter pylori is a common Gram-negative bacterium that inhabits the human stomach and causes a variety of gastric pathologies. One of the growing concerns is its dynamic spread of antibiotic resistance, a process in which biofilm formation is involved. Therefore, it is necessary to find an appropriate, high-throughput research model for the in vivo biofilm development by H. pylori. The aim of the current research report was to determine the usefulness of G. mellonella larvae in assessing the survival of a multidrug-resistant, strong biofilm producing H. pylori strain during its exposure to stress caused by clarithromycin. Using infection models lasting for 3 or 6 days, we confirmed the ability of the tested H. pylori strain to survive in the larvae. We noticed that exposure to clarithromycin significantly reduced the number of cultured bacteria relative to the control, although we did not observe any differences in the number of bacteria using time-lapse, live analysis of fluorescently stained larval hemolymph samples. In conclusion, we confirmed that the examined H. pylori strain can produce biofilm in G. mellonella larvae organism and is able to survive exposure to minimal inhibitory concentrations of clarithromycin (established in vitro) in in vivo conditions. Further refinement of methodologies for monitoring the viability of clinical H. pylori strains in the greater wax moth larvae will enhance the accuracy and reliability of this promising research model.

The present study describes the isolation and separation of isogarcinol and garcinol from kokum fruit by flash chromatography using water (containing 0.1% formic acid) and methanol mixed with acetonitrile (1:1) with UV detection at 254 nm and was characterised by HR-MS and NMR studies. These were further subjected to antimicrobial studies on Staphylococcus aureus FR1722 and Listeria monocytogenes Scott A by agar diffusion assay and broth microdilution method wherein the MIC (Minimum Inhibitory Concentration) of garcinol and isogarcinol were, 20 μg/mL and 50 μg/mL for S. aureus FR1722 and 50 μg/mL and 100 μg/mL for L. monocytogenes Scott A respectively. Further, studies on the control of biofilm growth were tested using MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide) assay followed by Crystal Violet (CV) assay, which showed up to 80% inhibition. In addition, the biofilm eradication as evaluated by bright field microscopy and the metabolic activity were also carried out against these bacterial strains and the experimental details are presented.

Leishmania is responsible for a neglected tropical disease affecting millions of people around the world and could potentially spread more due to climate change. This disease not only leads to significant morbidity but also imposes substantial social and economic burdens on affected populations, often exacerbating poverty and health disparities. Despite the complexity and effectiveness of the immune response, the parasite has developed various strategies to evade detection and manipulates host cells in favor of its replication. These evasion strategies start at early stages of the infection by hijacking immune receptors to silence critical cellular response that would otherwise limit the pathogen's propagation. Among these receptors, Fc receptors have emerged as a significant player in the immune evasion strategies employed by microorganisms, as they could promote inhibitory pathways. This review explores the potential role of one of these immune receptors, the FcγRIIA, in leishmaniasis and how this parasite may use it and the signaling pathways downstream to evade the host immune response. By understanding the potential interactions between Leishmania and immune receptors such as FcγRIIA, we may identify novel targets for therapeutic intervention aimed to enhance the host immune response and reduce the burden of this disease.

The complex interactions between parasites, their hosts, and associated microbiota hold significant implications for host health and disease outcomes. Helminths like Ascaris lumbricoides and Ascaris suum can significantly alter the host's intestinal microbiota, affecting both parasite biology and host pathology. Despite extensive research on host-microbiota changes due to helminth infections, the study of helminth-associated microbiota remains limited. This study aims to characterize the microbiota associated with Ascaris larvae and surrounding host tissues at distinct developmental stages (day 4, day 8, day 14), during larval migration through the liver, lungs, and intestine, and its impact on the host's microbiota in a murine model. Twenty mice were infected with 2500 embryonated A. suum eggs via oral gavage. Five Ascaris-infected mice and age-matched naïve mice were euthanized at 4-, 8-, and 14-days post-infection (DPI). Stool, intestine, liver, and lung samples were collected. Larvae were isolated from embryonated eggs in vitro, from the liver at 4 DPI, and the lung at 8 DPI. Utilizing 16S rRNA sequencing, we analyzed bacterial diversity in samples from different Ascaris stages and host tissues. Our results revealed a total of 8040 amplicon sequence variants (ASVs) with Ascaris samples displaying the highest diversity. Notably, Ascaris-larvae associated microbiota differed significantly from that of the host, with higher diversity observed in the parasite. Differential abundance analysis identified distinct taxonomic patterns, highlighting specific genera such as Bradyrhizobium, Achromobacter, and Pseudomonas in Ascaris. Our findings suggest that Ascaris harbors a unique microbiota that potentially exchanges bacteria with the host during larval migration. These insights pave the way for further research into the ecological and functional dynamics of helminth-microbiota interactions, which may inform novel therapeutic strategies targeting these microbial relationships to mitigate helminth infections and improve host health outcomes.

Current treatment of Candida infections is threatened by antifungal drug resistance. Thus, medicinal plants have been studied to identify new and highly effective antifungal substances with low toxicity. Here, we showed that the tannin-rich n-butanol fraction of Terminalia catappa (FBuOH) possesses antifungal and antibiofilm properties and protects Tenebrio molitor larvae against Candida albicans infection. FBuOH showed antifungal activity against Candida spp. vaginal isolates (MIC values ranged from 7 to 500 μg/mL). Moreover, a combination of FBuOH with fluconazole (FICI ≤ 0.5) showed considerably increased anti-yeast, anti-biofilm activity and significantly improved the survival rate (up to 100%) of T. molitor larvae against C. albicans infection. Furthermore, FBuOH acted synergistically with fluconazole by reducing C. albicans membrane ergosterol content. These results could also explain the synergistic activity between FBuOH and fluconazole, indicating that FBuOH exerted its effects on C. albicans membrane integrity, increasing its permeability. Our findings provide insights into the antifungal activity and low cytotoxicity of FBuOH, showing its potential use as a new antimycotic.

Mycobacterium tuberculosis (Mtb) is the bacterium responsible for causing Tuberculosis (TB) and understanding its mechanisms of virulence, persistence, and pathogenesis is a global research priority. Attenuated strains of Mtb are valuable tools for investigating the genes and proteins involved in these processes. In this study, we identified an Mtb mutant, H37Rv-S, which exhibits a shorter mycelium, smoother colony, slower growth, and reduced antibiotics resistance compared to the wild-type strain H37Rv. Genomic sequencing revealed 34 mutation events in the coding regions of H37Rv-S, affecting 31 genes. We conducted TMT-labeling quantitative proteomics to compare the expression differences between H37Rv-S and H37Rv, as well as their infected bone marrow derived macrophages (BMDMs). The results showed that 716 protein groups (23.96%) in H37Rv-S and 115 protein groups (2.99%) in the infected BMDMs were differentially expressed. The dysregulated proteins in H37Rv-S correspond with its phenotype characteristics. Among the 31 affected genes in H37Rv-S, 10 showed upregulation and 1 showed downregulation at the protein level. Notable, 16 associated network proteins in the phoP/phoR system were significantly dysregulated due to a frameshift mutation in phoP, altering its protein sequence after Phe45. The dysregulated host proteins in H37Rv-S were associated with immune response, necroptosis, and ferroptosis. Additionally, H37Rv-S demonstrated reduced survival capability in strain fluorescence labeling and colony-forming unit (CFU) counting post-infection of BMDM cells. These findings suggest that H37Rv-S is an attenuated strain exhibiting defective phenotype characteristics and is more readily recognized and eliminated by the host. This enhanced understanding of the differences between virulent and attenuated strains could facilitate the development of new targets and therapeutics for TB prevention and treatment.

Understanding Cryptococcus neoformans pathogenesis requires a detailed analysis of the various virulence factors that contribute to its ability to cause disease. Cyclosporine, calcineurin inhibitor, impairs C. neoformans production of a polysaccharide capsule and secretion of urease, which are critical for cryptococcal pathogenesis. Two particularly important virulence factors are the production of cell wall melanin and formation of biofilm. In this study, we investigated cyclosporine's effects on melanin production and biofilm formation in C. neoformans. Initially, we examined melanin production in planktonic cells treated with cyclosporine using an L-DOPA containing melanin-inducing medium. Visual inspection and optical microscopy revealed a notable reduction in the characteristic dark coloration of cultures treated with cyclosporine, which indicate decreased melanin production in daughter cells compared to mother cells. Spectrophotometric analysis also demonstrated a significantly altered ultraviolet-visible (UV/vis) absorption spectra in cyclosporine-treated yeast cells, indicative of structural changes in melanin. Additionally, cyclosporine-treated cells exhibited reduced conductance (P-value < 0.0001), suggesting altered cellular ionic properties. The impact of cyclosporine on biofilm formation and mature biofilm disruption was also assessed. Despite cyclosporine's efficacy in modifying virulence factors during planktonic growth, cyclosporine did not inhibit biofilm formation or melanization under biofilm growth conditions, nor did it disrupt mature biofilms in terms of biomass or metabolic activity. However, there was a significant reduction in extracellular matrix production in cyclosporine-treated non-melanized biofilms. Our findings underscore the complex interplay between cyclosporine and C. neoformans, highlighting its differential effects on melanization and biofilm dynamics, which provides new insights into the shortcomings of cyclosporin for combatting cryptococcosis and informs pathways for future therapeutic strategies against cryptococcosis.

The exposure risk of plasmid-mediated quinolone resistance (PMQR) genes increases the incidence of resistant bacterial infections, has resulted in clinical treatment failures with ciprofloxacin, necessitating urgent implementation of novel strategies for controlling this situation. Matrine serves as the principal constituent of the traditional Chinese herb Sophora flavescens Ait. and exhibits pharmacological activities including anti-inflammatory, antibacterial, anti-tumor, and hepatoprotective effects. However, the precise mechanism by which matrine exhibits antibacterial activity remains incompletely elucidated. This study investigated the antibacterial potential and synergistic mechanism of matrine in combination with ciprofloxacin against qnrS-carrying E. coli. The clinical ciprofloxacin-resistant E. coli carrying the qnrS and the recombinant E. coli DE3 (pET28a-qnrS) were evaluated for their antibacterial activity in vitro, it was found that the combination of matrine/ciprofloxacin exhibited a significant synergistic, reducing the MIC value of ciprofloxacin against qnrS-positive E. coli by 4-fold, and it effectively reduced the bacterial load to undetectable levels within 10 h without obvious cytotoxicity. Moreover, consistent findings were observed in significantly reducing bacterial load within the mouse infection model. Molecular docking revealed that matrine was localized in the large loop B of the qnrS crystal structure, establishing hydrogen bonds with Thr-102 and Arg-101, thereby disrupting the activity of qnrS. Interaction analysis further confirmed that matrine could significantly inhibit the protective effect of qnrS on gyrase and restore the activity of ciprofloxacin against qnrS-positive E. coli. Matrine may serve as a qnrS inhibitor to restore the efficacy of ciprofloxacin, suggesting its potential as a novel antibiotic adjuvant for controlling bacterial infections.

Plants contain a wide variety of bioactive compounds, which have attracted the interest of researchers in finding novel sources of natural medicine. In the following paper, we aim to evaluate the antibacterial potential of extract fractions associated with Parkia speciosa pods and beans against human pathogenic bacteria.

The majority of anti-leishmanial drugs used for treating trypanosomatid parasites help to reduce human morbidity and mortality. However, parasites have developed drug resistance, which has made it challenging to treat leishmaniasis. Therefore, new drugs and drug targets need to be identified. Protein synthesis is a crucial anabolic mechanism necessary for parasite survival. Histidyl-tRNA synthetase (HisRS) is an essential enzyme that is required for histidine incorporation into proteins. Recent studies on HisRS have shown differences between trypanosomatid HisRS and human HisRS, which could lead to the development of trypanosomatid HisRS structure-based inhibitors. This study aims to determine the role of L. donovani HisRS (LdHisRS) in parasite growth and virulence in vitro using RNAi. The silencing effect of LdHisRS expression was determined using qPCR. The results showed that after 24 and 48 hours of incubation with 90 ng siRNAs, LdHisRS mRNA expression levels were significantly reduced by ∼3.14-fold and ∼3.90-fold, respectively. SiRNA-treated parasites also exhibited ∼46.6% delayed growth and ∼47% reduced virulence. Additionally, homology modeling, virtual screening, and molecular docking studies were performed with potential inhibitors that have significant suppressive activity in bacteria, fungi, and viruses. . Halofuginone was found to have the best binding affinity of -9.09 kcal/mol as a potent inhibitor against LdHisRS. The molecular dynamics (MD) results showed that halofuginone could interact with the various active site segments, potentially blocking substrate access. The data on gene silencing through siRNA suggests that LdHisRS is essential for the parasite's growth and survival. The computational findings could lead to the development of a potent ligand (halofuginone) as a future anti-leishmanial drug, paving the way for an effective therapeutic treatment.

Brucellosis is a zoonotic bacterial disease that causes enormous economic losses in livestock populations and severe debilitation in humans globally. This study analyzes the seroprevalence of human brucellosis in Iran from 1970 to 2023, revealing key epidemiological trends based on data from 20,046 individuals. Through serological, culture, and molecular tests, it identifies the Brucella species affecting humans over five decades. The findings emphasize the need for ongoing monitoring and improved disease control efforts. Subsequently, we cultured specimens from seropositive individuals who exhibited suspected symptoms of brucellosis. The specimens included blood (n=9017), placentas (n=203), bone marrow (n=221), aborted fetuses (n=70), cerebrospinal fluid (CSF) (n=62), synovial fluid (n=5), urine (n=4), sperm (n=3), lumbar discs (n=3), abscesses (n=2), milk (n=2), and vertebral bodies (n=1). The overall seroprevalence of brucellosis in Iran was 44.9% based on RBPT, 20.7% by SAT, and 8.7% by 2-ME tests. The highest SAT prevalence occurred in the 2000s (25.8%), followed by the 1990s (23.8%), 1980s (22.2%), 1970s (14.8%), and 2010s (10.4%). Two main Brucella species were identified in infected humans as Brucella melitensis and Brucella abortus. Positive culture rates varied by sample type, with 9.9% for blood, 10.4% for bone marrow, and up to 100% for fluids like urine and milk. The most common strain was B. melitensis biovar 1 (89.2%), while B. abortus biovar 1 had a much lower incidence (0.3%). This investigation showed that the highest rates of culture positivity were in samples of urine, synovial fluid, abscess, and milk. These findings suggest that B. melitensis continues to be an important public health concern and that recent years of decline may be due to better control measures of the disease. The combined use of serology and culture is recommended for diagnosing brucellosis. Increased awareness among farmers and consumers of raw dairy products, coupled with nationwide control measures, is crucial for combating zoonotic brucellosis.

Coxiella burnetii (Cbu) is the gram-negative intracellular pathogen responsible for deadly zoonotic infection, Q fever. The pathogen is environmentally stable and distributed throughout the world which is sustained in nature by chronic infection of ruminants. The epidemiological studies on Q fever indicates it as emerging public health problem in various countries and it is imperative to promptly identify an appropriate therapeutic solution for this pathogen. In the current study, metabolic pathways of Cbu were analysed by the combination of multiple computational tools for the prediction of suitable therapeutic candidates. We have identified 25 metabolic pathways which were specific to Cbu containing 287 unique proteins. A total of 141 proteins which were either virulent, essential or resistant were shortlisted that do not show homology with the host proteins and considered as potential targets for drug and vaccine development. The potential therapeutic targets were classified in to seven functional classes, i.e., metabolism, transport, gene expression and regulation, signal transduction, antimicrobial resistance stress response regulator and unknown. The majority of the proteins were found to be present in metabolism and transport class. The functional annotation showed the predominant presence of proteins containing HATPase_c, Beta-lactamase, GerE, ACR_tran, PP-binding, CsrA domains. We have identified Type I secretion outer membrane protein for the design of multi-epitope subunit vaccine using reverse vacciniology approach. Four B cell epitopes, six MHC-I epitopes and four MHC-II epitopes were identified which are non-toxic, non-allergen and highly antigenic. The multi-epitope subunit vaccine construct was 327 amino acid residues long which include adjuvant, B cell epitopes, MHC-I epitopes and MHC-II epitopes. The Cholera enterotoxin subunit B is included as an adjuvant in the N terminal of vaccine construct which will help to produce a strong immune response to the vaccine. The multi-epitope vaccine construct was non-toxic, non-allergen and probable antigen having molecular weight 35.13954 kDa, aliphatic index 85.50, theoretical PI 9.65, GRAVY -0.001, and instability index of 28.37. The tertiary structure of the vaccine construct was modeled and physiochemical properties were predicted. After validation and refinement of tertiary structure the molecular docking of vaccine exhibited strong binding with TLR2, TLR3, TLR4, TLR5 and TLR8. The TLRs and vaccine construct formed hydrogen bonds, salt bridges and non-bonded contacts with all TLR receptors. The in-silico immune simulations showed the ability to trigger primary immune response as shown by increment in B-cell and T-cell population. The research paves the way for more effective control of zoonotic disease Q fever.

The gut microbiota of FUNAAB Alpha chickens plays a crucial role in determining their overall health and performance. Understanding the various types and diversity of microbiota in the gut of different genotypes of chickens is crucial for enhancing their well-being, productivity, and disease resistance. This study employed 16S rRNA and metagenomics analysis to examine the gut bacteria of three genotypes of FUNAAB Alpha broiler chickens, namely Naked neck, Frizzle, and Normal feather. There were three phyla observed in the three genotypes: Bacteroidetes, Actinobacteria, and Firmicutes. Through the utilisation of 16S rRNA sequencing, we successfully identified and categorised the various microbiota present within the gastrointestinal tract. Our study revealed notable variations in the composition and quantity of microbiota across the three genotypes, suggesting that each genotype possesses a distinct collection of gut bacteria. A wide range of microbiological diversity was observed within the community. Interestingly, the Normal feather chicken exhibited a greater number of operational taxonomic units (OTUs) compared to the Naked neck and Frizzle feather chicken. This study presents significant findings regarding the ceacal microbiota of FUNAAB Alpha chickens, emphasising the variations observed among different genotypes. It is crucial to study composition-modification techniques to enhance host health and performance, as well as to control zoonotic pathogens that can contaminate poultry products and threaten consumers' health.

Camel contagious ecthyma is a highly infectious viral skin disease that affects camels and causes economic losses. This study reports the isolation and the phylogenetic analysis of contagious ecthyma virus among camels in Morocco. The disease was detected in four among fifteen camels with severe papules on the lips and nares. Samples of skin crusts were collected and pooled for virus isolation and titration, PCR testing, and histopathological examination. PCR was used to amplify the B2L gene and the resulting product was sequenced and analyzed genetically. The study's findings indicate the presence of characteristic microscopic changes of poxvirus infection in the examined tissues and the virus was isolated on two cells (lambs testis and Vero) and showed distinct growth patterns. The virus grew rapidly on TA cells and delayed growth on Vero cells. The third passage showed cytopathic effects characterized by cell aggregation. Sequence analysis of the B2L gene revealed 100% similarity to the camel contagious Ecthyma virus isolated from Ethiopia. The camel virus isolates can be classified into two genetic clades according to the B2L gene sequence: the Asian lineage, which includes isolates from Saudi Arabia, Bahrain, and India, and the African lineage, which includes isolates from the Sudan. In conclusion, this is the first instance of the camel contagious ecthyma virus being identified in North Africa in a herd of camels following exposure to stress. Moreover, the progression of the disease was closely monitored from onset to recovery in a setting without the bacterial complication often observed in the field. The virus exhibited opportunistic behavior, exploiting the stress response. Further studies are warranted to evaluate the pathogenicity of the virus in camels and to genetically characterize the circulating virus from different regions. This will be highly beneficial in the development of an appropriate vaccine.

Colistin, a polymyxin antibiotic, serves as a crucial defense against multidrug-resistant gram-negative bacteria, despite its nephrotoxicity. However, the plasmid-mediated mobilizion of the polymyxin resistance gene, mcr-1, presents a significant public health threat. The widespread use of disinfectants has resulted in Escherichia coli (E. coli) carrying mcr-1 also showing disinfectant resistance. The aim of this study is to investigate the distribution of disinfectant genes and resistance to disinfectants in mcr-1-carring E coli from children in the South China.

According to many investigations, persistent oral infections may be caused by oral pathogenic biofilms. Irritation of soft tissues and subsequent bone resorption due to bacterial biofilm contamination of the implant further worsen oral health. Dental problems may be effectively treated using metal nanoparticles (NPs) because they limit the development of many different types of bacteria. With their low toxicity, X-ray sensitivity, high atomic number, near-infrared driven semiconductor qualities, and cheap cost, multifunctional bismuth (Bi) NPs with therapeutic activities show significant potential for the domains of bacterial infection diagnostics and treatment. Also, by directly communicating with the bacterial cell wall, stimulating intracellular effects, inhibiting biofilm formation, producing reactive oxygen species, and inducing adaptive and innate immune responses, BiNPs offer an alternative to conventional antibiotics for treating bacteria with multiple drug resistance (MDR). Hence, BiNPs, which have more antibacterial activity and fewer side effects than chlorhexidine, might be a promising option to fight biofilm-forming bacteria in the mouth. This could lead to their usage in several areas of dentistry. The research delves into the many synthesis techniques of BiNPs and their antibacterial and anticancer capabilities. Next, we'll review how this nanoparticle has helped with dental infections, periodontitis, and dental implant problems. The anticancer effects of BiNPs on oral cancer were also studied. Thus, after this paper, we have highlighted the therapeutic limits and ways to address this issue for the clinical success of BiNPs in promoting oral and dental health.

Leptospirosis is a globally important re-emerging zoonotic disease affecting humans and many animal species including dogs. The present cross-sectional study aimed to diagnose acute leptospirosis among 210 suspected dogs using microscopic agglutination test (MAT) and polymerase chain reaction (PCR). Further, epidemiological risk factors were analyzed by univariate analysis and multivariate binomial logistic regression analysis. Out of the 210 dogs, anti-leptospiral antibody was detected in 123 (58.57 %; 95 % CI: 51.6-65.3) dogs by MAT (cut-off titer- ≥1:100), among which 67 (54.47 %; 95 % CI: 45.3-63.4) were seropositive to more than one serogroup. The predominant serogroups were Australis and Autumnalis in the study region. Whereas, acute leptospirosis was established in 49 (23.33 %, 95 % CI: 17.9-29.8) dogs based on ≥ 1:800 titer in unvaccinated dogs and ≥1:1600 titer in vaccinated dogs. The predictive risk factors of acute canine leptospirosis were adult dogs (1-5 years) (p = 0.001), north-east monsoon season (p = 0.011), outdoor management (p = 0.047), history of rodent exposure (p = 0.001) and history of contact with wet soil or stagnant water (p = 0.046). Among 49 acutely infected dogs, thirteen dogs were positive for urine PCR and one dog was positive for both urine as well as plasma PCR. Positive PCR amplicons were identified as Leptospira interrogans based on secY gene sequencing and phylogenetic analysis. This study enlightened about the occurrence of acute leptospirosis among suspected dogs with the above important predictive risk factors, which should be taken into consideration while history taking so that proper treatment strategies can be adopted for early recovery of the animal.

Aeromonas hydrophila is a significant pathogen in the field of fish farming, resulting in substantial financial losses for the aquaculture industry. As the pathogen's resistance to commercially available antibiotics continues to rise, the identification of novel antimicrobial strategies becomes increasingly crucial. This study aims to explore the modulatory impact of gsk-3β (Glycogen synthase kinase-3β) on the intrinsic immunity against Aeromonas hydrophila in zebrafish, with the objective of uncovering a new avenue for enhancing fish antimicrobial activity through gene editing. Our investigation involved an analysis of the evolutionary patterns and protein sequence of gsk-3β, elucidating its conserved characteristics in zebrafish and fish species of economic importance. In this research, CRISPR-Cas9 technology was employed to generate a zebrafish model with a knockout of gsk-3β, resulting in a decreased resistance of zebrafish to Aeromonas hydrophila (ATCC 7966) infection. Furthermore, we conducted preliminary investigations into the potential mechanisms through which gsk-3β governs antimicrobial immunity. Our findings revealed that knockout of gsk-3β resulted in diminished activation of innate immunity, antioxidant capacity, and autophagy. Hence, the findings of this study are highly significant in improving the economic benefits of aquaculture and in effectively preventing and controlling infection caused by Aeromonas hydrophila.

Isoniazid stands as a frontline antibiotic utilized in the treatment of tuberculosis (TB), predominantly impacting the mycolic acid component within the cell wall of Mycobacterium tuberculosis (Mtb). It also affects the formation of lipoarabinomannan (LAM), an essential glycolipid in the cell envelope of Mtb. Despite the effectiveness of antibiotics for TB treatment, drug tolerance development in mycobacteria frequently stems from their adaptation to the hostile environment within the host, leading to treatment failure. Herein, we investigate mycobacterial adaptation to the isoniazid exposure in the host-mimicked conditions by focusing on the stress response genes (virS, icl1, whiB3, tgs1) and LAM-related genes (lprG, p55, lmeA, mptA, embC). Mtb H37Rv and mono-isoniazid resistant (INH-R) strains were cultivated in the host-mimicked multi-stress condition (MS) with or without isoniazid and the relative expressions of these gene candidates were measured using real-time PCR. In the INH-R strain, treatment with isoniazid in multi-stress conditions caused significant upregulation of tgs1, and LAM precursor-lipomannan (LM) synthesis and its transport genes (lprG, p55, lmeA, embC). In the case of H37Rv, all LAM-related genes and tgs1 were downregulated whereas other stress response genes were upregulated, remarkably in icl1 and whiB3. These findings highlight differences in gene expression patterns between drug-sensitive and resistant strains in multi-stress environments with drug pressure. Notably, stress response genes, particularly tgs1, may play a crucial role in regulating LAM production in the INH-R strain in response to isoniazid exposure. This study enhances our understanding of the mechanisms underlying drug resistance, offering valuable insights that could contribute to the development of new strategies for treating and eliminating TB.

Staphylococcus pseudintermedius is an opportunistic inhabitant of skin and mucosal membranes in animal species. Its resistance to β-lactam antibiotics is usually measured by β-lactamase enzyme production. This study was aimed to identify S. pseudintermedius from broilers, determine the prevalence of blaZ gene, and evaluate the relationship between agr type and β-lactamase gene expression. Fifty swabs from nasal cavities of broilers were sampled. To identify S. pseudintermedius, the nuc gene was amplified by PCR. Antibiotic susceptibility testing was performed on the isolates by the Kirby Bauer technique, and the presence of blaZ gene was detected. Finally, the relative gene expression of blaZ was determined in each agr group. Results of gene amplification showed that 33 samples (66%) contained nuc gene of which 21 samples horbored blaZ and were resistant to penicillin (%63). The Multiplex PCR was used to determine the agr type of blaZ positive isolates. Of the strains, 70% were categorized in the agr I group, 21 % in the agr II group, 0% in the agr III group, and 9 % in the agr IV group. Results of relative gene expression of blaZ with gyr B as a housekeeping gene showed that the change fold of the blaZ gene was significantly higher in the agr I group compared to the other groups (p≤0.05). These data suggested that agr activity may influence the resistance of S. pseudointermedius to β-lactam antibiotics. In this study, S. pseudintermedius was isolated for the first time from the nasal cavities of broilers.