Search Results (480)

Chicken coccidiosis is a parasitic disease caused mainly by Eimeria tenella, Eimeria acervulina, Eimeria maxima, and Eimeria necatrix, with most cases presenting as mixed infections. Currently, although a subunit vaccine (CoxAbic) targeting Eimeria maxima via maternal immunization is commercially available, no genetically engineered multivalent subunit vaccine exists against mixed infections caused by these four Eimeria species simultaneously. Therefore, we developed a tetravalent recombinant subunit vaccine (designated TEIN) by fusing key antigen genes (TA4, 3-1E, IMP1, NA4) from these four Eimeria species and expressing the construct in Pichia pastoris. A total of 500 chickens were randomly allocated into 25 experimental subgroups (n = 20 each), consisting of five groups (control, challenged, adjuvant, pPIC9K, and TEIN) and five challenge conditions (infection with Eimeria tenella, Eimeria acervulina, Eimeria maxima, Eimeria necatrix, or a mixture of four species). Immunization was performed via leg intramuscular injection at 14 and 21 days of age. At 28 days of age, all chickens except the controls were orally challenged with 1 × 10⁴ sporulated oocysts. Statistical analysis was performed using one-way or two-way ANOVA as appropriate. Results showed that chickens vaccinated with the TEIN subunit vaccine exhibited significantly elevated serum levels of IgY, IL-2, IL-10, and IFN-γ, as well as an increased splenic lymphocyte CD4⁺/CD8⁺ ratio. The anticoccidial indices (ACI) against Eimeria tenella, Eimeria acervulina, Eimeria maxima, and Eimeria necatrix, and their mixed infection reached 174.82, 174.58, 174.41, 180.61, and 175.95, respectively. Moreover, no significant differences were observed in hematological parameters, serum biochemical markers, or histopathological findings between the vaccinated and control groups. These results demonstrate the vaccine’s potential as a promising candidate for controlling mixed coccidial infections. Full article

Botulinum toxin type A (BoNT/A) injection into the bladder wall is a milestone in the treatment of urinary incontinence in patients with neurogenic detrusor overactivity (NDOi) or overactive bladder syndrome (OABi) who are refractory to or unable to tolerate oral or transdermal therapies. However, the efficacy of BoNT/A is hampered by the low long-term adherence of patients to a treatment that requires repeated bladder injections under cystoscopy control. The discontinuation is particularly evident among incontinent patients with spontaneous voluntary voiding, regardless of whether the cause is NDOi or OABi, although clearly more marked among the latter group. In addition to the bother and pain associated with repeated cystoscopies, these patients show low tolerance to the high incidence of urinary tract infections (UTIs) and transient urinary retention, the two most common adverse events. Fewer injection points may render treatments less painful, apparently without reducing efficacy, but will not avoid the need for repeated cystoscopies, and no studies have demonstrated that such modification increases adherence. Eventually, accessing the bladder wall for BoNT/A administration via a transabdominal approach, under real-time ultrasound guidance, may overcome trans-urethral limitations, but the technique’s reproducibility remains unknown. An intensive investigation is ongoing to identify aids that facilitate the passage of the large, fragile BoNT/A molecule across the urothelium to reach the bladder nerves without injections. Electromotive Drug Administration (EMDA) of BoNT/A demonstrated efficacy and safety over a 6-year follow-up in NDOi patients at a single center, but the results were not reproduced at other institutions. The application of shock waves to the bladder using shock waves generated by Extracorporeal Shock Wave Lithotripsy (ESWL) machines to tear the urothelium and facilitate the passage of BoNT/A instilled in the bladder is ingenious, but the experience is very limited. Dimethyl sulfoxide, liposomes, and thermal-reversal hydrogel to deliver the toxin failed in pilot trials. BoNT/A in nano-formulations has high heat stability, resistance to pH changes, and to enzymatic degradation. Extended efficacy in dermal and intramuscular pilot applications is promising but needs to be replicated in the bladder. Full article

Background: Global immunization programs continue to rely on needle-based injections despite persistent concerns regarding sharps disposal, accidental injuries, and the technical skill required for accurate intradermal administration. Needle-free jet injectors (NFJIs) are an alternative delivery method in which narrow, high-velocity liquid jets penetrate the skin without a needle. Contemporary designs, ranging from single-use disposable-syringe injectors to digitally controlled electromechanical devices, address historical safety issues and meet current WHO and FDA device expectations. Methods: Evidence from engineering analyses, preclinical modeling, and clinical trials was reviewed to characterize how jet velocity, nozzle structure, and formulation rheology influence skin penetration and drug dispersion. Published vaccine studies were examined for antibody responses, seroconversion, and reactogenicity compared with needle–syringe injection. Field vaccination campaign data from national campaigns and operational reports were evaluated to describe implementation steps, acceptability, and implementation constraints. Results: Published studies evaluating vaccines, including inactivated influenza, hepatitis B, typhoid, rabies, and measles, report antibody titers and seroconversion rates after NFJI administration that are comparable to those achieved with conventional intramuscular or intradermal needle injection. Needle-free delivery was associated with operational advantages in several immunization programs, including reduced sharps waste and improved vaccination rate during high-volume immunization campaigns. Local and systemic reactogenicity follows expected patterns, with slightly higher injection-site responses in some NFJI studies. Imaging and mechanical data confirm that jet performance depends on nozzle geometry and controlled pressure pulses. At the same time, formulation stability remains a critical determinant of successful jet-based vaccine administration, particularly for protein antigens, adjuvanted formulations, and emerging mRNA vaccines that may experience transient shear stress during high-velocity injection. Evidence from vaccination campaigns further indicates that needle-free jet injectors reduce sharps waste, simplify vaccine handling and administration procedures, and support rapid vaccine delivery in large-scale immunization programs. Conclusions: Needle-free jet injectors are a practical alternative to traditional needle-based injections for some vaccines. Their main benefits include enabling intradermal dose-sparing strategies, reducing reliance on sharps disposal methods, and enabling the efficient vaccination of large groups without compromising immunogenicity. Future research should define the physicochemical stability limits of biologic formulations subjected to jet injection and evaluate digitally controlled injectors capable of precise pressure modulation and adjustable delivery parameters. In addition, needle-free jet injection eliminates needle penetration and sharps handling, which may reduce needle-associated anxiety and improve vaccine acceptability among individuals with needle aversion. Full article

Background: Chronic sclerosing osteomyelitis of Garré (CSO) is a rare, non-suppurative form of primary chronic osteomyelitis characterized by reactive periosteal bone formation and cortical thickening. It most commonly involves the mandibular bones, whereas long-bone localization is uncommon. Material and Methods: We report a 4-year-old girl who developed progressive right thigh pain and limping six months after receiving intramuscular ampicillin injections. Subsequent evaluation revealed femoral changes consistent with chronic sclerosing osteomyelitis. Surgical decompression and targeted antimicrobial therapy were performed. Results: Microbiological analysis of intraoperative specimens obtained prior to antibiotic therapy yielded Staphylococcus epidermidis (S. epidermidis) and Staphylococcus capitis (S. capitis). After three years of follow-up, the patient exhibited no functional impairment or growth disturbance of the affected limb. Conclusions: Although coagulase-negative staphylococci (CoNS) are commonly regarded as skin commensals, their repeated isolation from deep surgical specimens, together with clinical findings and response to treatment, raises the possibility of their involvement in the disease process in this case. Full article

Background/Objectives: Bacteriophage-based display has been utilized for a variety of purposes, such as to assemble protein libraries and conduct biopanning. We have created a modified lambda (λ) bacteriophage platform, ideal for the display and delivery of proteins. Our system utilizes counter-selection recombineering for versatile modification, temperature-sensitive induction for timely lysate production, and an arabinose-inducible mechanism for high-titer, stable yield. Here, we investigated the ability of this specialized λ phage display platform to stimulate highly specific antibodies in mice against the displayed cancer-variant cell-surface receptor EGFRvIII, demonstrating its potential in cancer immunotherapy and broader vaccine development. Methods: λ display immunogenicity was explored by generating fusion proteins between the λ head protein D and a 13-mer peptide from the N terminus of glioblastoma variant cell-surface receptor, EGFRvIII. The 13-mer peptide was fused to either the N or C terminus of the λD protein while λ remained a dormant lysogen in the bacterial host chromosome. Recombinant phage lysates were then generated with ~420 displayed fusion proteins per phage particle. Mice were injected with purified recombinant λ phage without an adjuvant via both intraperitoneal and intramuscular routes, and sera harvested at various timepoints were profiled for immunogenicity. Results: Analysis of serum samples by ELISA and Western blotting demonstrated the ability of the λD~EGFRvIII phage display, especially in the C-terminal fusion construction, to elicit a robust anti-EGFRvIII humoral response by either injection route. Notably, the antibody response was highly specific to EGFRvIII without exhibiting cross-reactivity to wild-type EGFR. Conclusions: The data generated in this study demonstrate the λ system’s immunotherapeutic potential as a high-titer, stable, self-adjuvanting vector for the stimulation of robust antibody titers with defined specificity. Full article

Lekethromycin (LKMS) is a novel macrolide veterinary antimicrobial. Its propensity for intracellular accumulation causes discrepancies between whole blood and plasma concentrations, complicating pharmacokinetic evaluations. This study compared the pharmacokinetic characteristics, dose proportionality, and bioavailability of LKMS in whole blood and plasma following intramuscular administration in pigs. Forty-two healthy pigs received LKMS via a single intravenous reference dose (5 mg/kg) for absolute bioavailability estimation or intramuscular (1, 2.5, 5, and 10 mg/kg) injection. Pharmacokinetic parameters were calculated using non-compartmental analysis, and dose proportionality was evaluated via a power model. LKMS exhibited rapid absorption and slow elimination, with a plasma half-life of 49.25 to 67.63 h. Whole blood exposure and peak concentrations were 1.5 to 3 times higher than in plasma, indicating extensive blood cell partitioning. As the intramuscular dose increased, the whole blood-to-plasma concentration ratio decreased from 2.83 to 1.15, suggesting a saturable cell uptake mechanism. Consequently, LKMS exhibited non-linear pharmacokinetics in whole blood but demonstrated linear, dose-proportional pharmacokinetics in plasma. Absolute bioavailability based on plasma ranged from 83.2% to 119.5%. Due to saturable blood cell binding, plasma is the optimal matrix for accurately evaluating LKMS systemic exposure and bioavailability in swine. Full article

Background: The global spread of monkeypox virus (MPXV) highlights an urgent need for thermostable and easily administrable vaccines. Current orthopoxvirus vaccines are limited by cold-chain dependence and inconvenient injection-based delivery. Objectives: This study aimed to develop a dissolvable microneedle (DMN) vaccine against monkeypox based on a replication-deficient orthopoxvirus platform, through systematic formulation screening, stabilization mechanism exploration, and rigorous in vivo immunogenicity evaluation. Methods: A film-based approach was adopted for efficient, high-throughput formulation screening and thermostability assessment. NTV was mixed with excipients and dried into solid films. Stability was monitored via RT-qPCR after storage at 4 °C to 40 °C. The lead formulation was physically characterized, then used to fabricate MVA-BN-loaded DMN patches, which were further evaluated for in vivo immunogenicity via immunization in BALB/c mice. Results: The optimal formulation F2 (containing dextran, L-threonine, and BSA/HSA) showed a potency loss of only ~1 log₁₀ after 2 months at 25 °C, and <1 log₁₀ loss after 1 week at 37 °C. SEM revealed a porous virus-entrapment morphology, and FTIR indicated enhanced hydrogen bonding between the virus and the dextran matrix. The formulation was successfully manufactured into DMNs that dissolved within 5 min. In mice, these DMNs elicited robust MPXV-specific IgG and neutralizing antibody responses, with immunogenicity comparable to that induced by conventional intramuscular injection. Conclusions: This study successfully established a thermostable formulation and dissolvable microneedle delivery platform for replication-deficient orthopoxvirus vaccines against monkeypox. The optimized DMN vaccine induced robust MPXV-specific immune responses in mice with immunogenicity comparable to intramuscular injection, addressing the core limitations of current vaccines and providing a promising solution for monkeypox prevention. Full article

Antibiotic residues in water buffalo milk are a food-safety concern, yet depletion data are scarce. The purpose of this study was to characterize the depletion profiles of amoxicillin (AMOX) and its two major metabolites, amoxicilloic acid (AMA) and amoxicillin diketopiperazine-2′,5′-dione (2,5-DKP), in Anatolian water buffalo milk after a single intramuscular administration and to estimate a milk withdrawal time relative to the EU MRL. We tested the hypothesis that AMOX concentrations would decrease below the EU MRL over successive milkings and that AMA and 2,5-DKP would exhibit depletion kinetics distinct from the parent compound. Five lactating Anatolian water buffaloes received a single intramuscular injection of amoxicillin (15 milligrams per kilogram). Milk was collected at each milking (twice daily) for seven days and analyzed by liquid chromatography–tandem mass spectrometry with quantification limits below the European Union maximum residue limit for amoxicillin in milk (4 micrograms per kilogram). Amoxicillin peaked at the second milking (mean 13.65 micrograms per kilogram), mean concentrations fell below the maximum residue limit from the sixth milking, and they became non-quantifiable from the tenth milking onward. Two major metabolites, amoxicillinic acid and amoxicillin diketopiperazine-2′,5′-dione, peaked earlier (2,5-DKP Tmax 12 h) or at higher concentrations (AMA Cmax 32.64 µg/kg vs. AMOX 13.65 µg/kg) and remained detectable up to the thirteenth milking, with longer apparent terminal half-lives (32.0 and 52.8 h) than amoxicillin (23.5 h); the mixed-effects model confirmed different depletion rates among analytes (milking × analyte interaction p = 4.63 × 10⁻⁵). A log-linear withdrawal model applying the EMA 95/95 tolerance limit indicated that the first time point at which the upper tolerance limit fell below the EU MRL was 84.7 h after dosing; rounded up to the next 12 h milking interval, this corresponds to a reported withdrawal period of 96 h (≈8 milkings). These results provide species-specific residue kinetics for amoxicillin in Anatolian buffalo milk and support considering metabolites in monitoring and withdrawal-time decisions. Full article

The transition period in dairy cows is marked by metabolic, oxidative, and immune challenges that increase susceptibility to periparturient diseases. Injectable mineral supplementation (IMS) has been proposed to support immunometabolic adaptation by enhancing antioxidant capacity and immune function, with consistent associations with improved health outcomes but variable effects on production. Therefore, this study evaluated the effects of repeated intramuscular multi-mineral supplementation during the transition period on health, metabolic stress, immune status, and productive performance in Holstein cows. Supplementation was associated with lower odds of subclinical hypocalcemia on day 4 postpartum in primiparous cows (p = 0.02) and overall for persistent subclinical hypocalcemia (p = 0.03). Multiparous cows (p = 0.04) and the overall population (p = 0.01) showed consistent effects on metritis following IMS. Supplemented cows had improved metabolic and uterine health indicators without affecting energy metabolism-related disorders. Although no differences were detected for major postpartum health disorders, its main benefits may involve immune competence, oxidative regulation, and physiological resilience rather than energy balance. Full article

Background: Facial expressions of pain are essential for pain assessment, yet subjective pain reports often vary between sexes. Traditional self-report measures are prone to bias, and objective methods are needed for more reliable pain evaluation. Objective: To develop and validate a subjectivity-free automated tool to assess acute low back pain using facial expressions recorded during a functional spinal extension task. Participants: Thirty healthy adults, aged 18–40 years. Methods: Participants received intramuscular injections of hypertonic (pain) and isotonic (placebo) saline in the lumbar region during separate sessions. Facial expressions were video-recorded during a submaximal lumbar extension task and analyzed using a custom software based on Haar Cascade and Local Binary Pattern Histogram algorithms, which are techniques that do not require neither training data nor subjective labeling, contrary to what happens in deep learning solutions. Results: The tool successfully detected significant differences in facial expressions between pain, placebo, and pain-free conditions (p < 0.001). Test–retest reliability was good (ICC = 0.85). While both sexes showed similar facial expression patterns during pain, males reported higher pain scores on the numeric rating scale (p < 0.01). Pain significantly reduced steadiness of force in both sexes. Conclusion: The automated tool objectively quantified facial expressions associated with acute low back pain and revealed sex-related differences in subjective pain perception. This multimodal approach integrating expression analysis, physical performance, and self-report may enhance the accuracy of pain assessment in physiotherapy settings. Full article

Background: In South Korea, foot-and-mouth disease (FMD), a highly contagious viral infection that affects cloven-hoofed animals, has led to the implementation of a bivalent FMD vaccination program. The current FMD vaccination strategy involves intramuscular (IM) administration to the shoulder region of the swine. However, this method is associated with adverse reactions at injection sites. Our previous studies have demonstrated that intradermal (ID) vaccination eliminates these side effects while maintaining immunogenicity comparable to that of IM vaccination. This study aimed to assess the early immune response induced by ID vaccination and compare its protective ability against FMDV serotype O with that of a commercial IM vaccine recently used in South Korea. Methods: An ID FMD vaccine was evaluated using two adjuvants, ISA 207 (50%) and EMULSIGEN-D (15%). Virus neutralization (VN) titers and structural protein levels were measured to compare efficacy across groups. To assess the early protective efficacy of ID vaccination, viral challenge experiments were conducted at 7 and 14 days post-vaccination (dpv). Results: Swine vaccinated via the ID route exhibited no clinical symptoms at 14 dpv, indicating effective early protection against FMD (O/AS/SKR/2019). In addition, no side effects of FMD ID vaccination were observed. Conclusions: These results suggest that ID vaccination could serve as a viable alternative to conventional IM vaccination, which is frequently associated with adverse effects. Importantly, this study demonstrates that ID vaccination can provide effective early protection within 7–14 days post-vaccination, highlighting its potential utility for emergency outbreak control. Full article

Florfenicol (FFC) is widely used to treat bacterial infections in veterinary medicine; however, its pharmacokinetic characteristics in reptiles remain limited. This study investigated the pharmacokinetic profiles of FFC after intramuscular (IM) injection at doses of 20 or 30 mg/kg body weight (b.w.) in freshwater crocodiles (Crocodylus siamensis). A sample of 10 healthy crocodiles was randomly divided into two groups (n = 5 for each group) according to a parallel study design. Blood samples were obtained from pre-dose to 168 h post-administration. Plasma FFC concentrations were quantified using high-performance liquid chromatography with diode array detection (HPLC-DAD) and analyzed by non-compartmental analysis. The mean maximum plasma concentrations of FFC were 4.05 µg/mL and 6.11 µg/mL for the 20 and 30 mg/kg b.w. doses, respectively. The mean elimination half-lives of FFC were long but not significantly different (51 h). The average plasma protein binding was 37.15%. Based on the pharmacokinetics/pharmacodynamics (PK/PD) index, a single dose of FFC via IM elicited plasma concentrations above the MIC₉₀ values reported for several susceptible bacterial pathogens. Consequently, both dose levels provided plasma exposure consistent with previously reported reference MIC values. However, further PK/PD and multiple-dose investigations are needed to refine species-specific dosage regimens. Full article

Background: Respiratory syncytial virus (RSV) remains a major etiologic agent of acute lower respiratory tract infection (ALRTI). Currently licensed RSV vaccines are administered by intramuscular injection and induce limited immunity at the respiratory mucosal interface, underscoring the need for effective mucosal vaccination strategies. Methods: To enhance mucosal immune responses, we used prefusion F protein (Pre-F) as the antigen and performed intranasal immunization in BALB/c mice. Four mucosal adjuvants (CpG-ODN, CTA1-DD, IFN-α, and PEI) were systematically compared across different dose levels to evaluate their immunological and protective efficacy. Results: Both adjuvant type and dose helped shape the magnitude and quality of the immune response and the level of protection. CpG-ODN showed a dose-restricted immunopotentiating effect: an intermediate dose (10 µg) significantly increased neutralizing antibody titers and nasal mucosal IgA responses, improved post-challenge body weight recovery, and reduced lung viral load, whereas higher doses provided no additional benefit and were associated with aggravated lung pathology. PEI and IFN-α exhibited dose-dependency within a certain range, but increasing doses did not result in further improvements in immune responses or protection; an intermediate dose (10 µg) was sufficient to elicit robust systemic and mucosal immunity. CTA1-DD improved selected immune parameters at appropriate doses, yet its overall immunopotentiating effects remained modest. Direct comparative analysis using the representative doses selected from the three dose levels for each adjuvant indicated that 10 µg CpG-ODN or PEI provided superior immunogenicity and protection, whereas PEI induced a Th2-biased immune profile at both humoral and cellular levels. Conclusions: These findings highlight that favorable immunogenicity and protection are achieved within defined dose windows rather than at maximal doses. Among the adjuvants studied, low-to-intermediate doses of CpG-ODN, particularly 10 µg, show strong potential for intranasal mucosal immunization with recombinant RSV Pre-F protein. By systematically comparing dose–effect profiles across multiple mucosal adjuvants, this study offers comparative insights into adjuvant selection and dose selection for intranasal RSV vaccine development. Full article

Introduction: Reliable identification regarding interfascial spaces proves essential to achieve successful nerve block analgesia; however, ultrasound guided approaches are recognized as challenging, particularly in obese or pediatric patients. In prior cadaveric and clinical investigations, multiple approaches were evaluated to identify methods for measuring injection pressures as a function of needle position relative to fascia. Our previous study proposed simpler method of finding interfascial spaces with the needle tip. In this study, it was examined whether needle tip design influences injection pressures during regional anesthesia procedures, via an ex vivo pig specimen setup. Methods: A bespoke apparatus for tracking injection pressure was deployed to enable continuous measurement of intraluminal pressure generated while delivering saline throughout ultrasound guided peripheral nerve block needles conducted within pig thigh specimens. Delivery was performed using an infusion pump. Three types of needles of the same manufacturer (Pajunk) and same diameter (22G) but with different tips (Facet, Facet S and Sprotte) were used to measure injection pressures during penetration through tissues until interfascial plane hydrodissection was created. Statistical analyses were performed to compare pressure levels, variability, and temporal pressure trends. Results: Ninety ultrasound guided injections in porcine thigh tissue were analyzed, with thirty procedures per needle type. Injection pressure differed significantly between intramuscular, fascial puncture, and interfascial phases, showing a distinct puncture peak (p ≤ 3.44 × 10⁻¹⁴). Needle geometry significantly affected pressures across all phases (Kruskal–Wallis intramuscular p = 2.0 × 10⁻⁶, puncture p = 7.52 × 10⁻⁸, interfascial p = 9.2 × 10⁻⁵), with large pairwise effects (Hedges g up to 1.51). The classical tip produced the highest intramuscular and higher interfascial pressures, the sharp tip required the lowest puncture pressure, and the lateral tip yielded the lowest intramuscular and interfascial pressures. Conclusions: Needle tip geometry substantially influences pressure dynamics throughout the injection process, with the classical design associated with the highest fascia-penetration injection pressures and the sharp needle exhibiting the lowest, while the lateral design associated with lowest intramuscular-penetration and interfascial pressures. Full article

Background/Objectives: Chronic heat stress impairs lipid mobilization from adipocytes, which reduces substrate availability for muscle metabolism. Systemic inflammation is a key facilitative response to heat stress, and we sought to determine if mitigating inflammation in heat-stressed wether lambs would improve lipid flux. Methods: Two cohorts of commercial feedlot lambs were heat stressed for 30 days. In study 1, heat-stressed lambs received dexamethasone injections every 3 days, fish oil capsules twice daily, or no intervention. In study 2, heat-stressed lambs received daily boluses of ω-3 polyunsaturated fatty acid Ca²⁺ salts (ω-3 PUFA) or no intervention. Results: In both studies, heat stress reduced ex vivo epinephrine-stimulated free fatty acid and glycerol mobilization from visceral adipose tissue. These deficits were partially resolved by fish oil and fully resolved by ω-3 PUFA. In study 1, fish oil recovered heat stress-induced deficits in circulating triglycerides and HDL-cholesterol but not in circulating free fatty acids. Fish oil and dexamethasone resolved the increase in muscle PPARα, indicating less lipid utilization for metabolism. In study 2, ω-3 PUFA resolved heat stress-induced deficits in muscle CD36 and PPARγ, indicating improved lipid uptake capacity. However, interventions did not resolve reduced intramuscular lipid content in either study. Conclusions: We conclude that inflammation was a primary facilitator of impaired lipid mobilization in heat-stressed lambs but was not the sole driver of lipid dysregulation. Nevertheless, targeting inflammation was a beneficial strategy for improving lipid flux during chronic heat stress. Full article