Avian metapneumovirus (aMPV) is a virus in the Pneumoviridae family associated with respiratory and reproductive disease in commercial poultry. In fall of 2023, the United States experienced an outbreak of subtypes A and B in commercial poultry flocks that has persisted through 2025. This case series report describes epidemiologic, clinical, and pathologic aspects of the cases submitted to the Pennsylvania Animal Diagnostic Laboratory System (PADLS) from January 10, 2024, to January 10, 2025. Of the 883 PADLS cases that requested aMPV PCR and/or ELISA testing during the study period, 405 were positive Pennsylvania cases: 169 on PCR, 221 on ELISA, and 15 on both PCR and ELISA. Lancaster County, which is the state's most agriculturally dense area, had the first reported aMPV case in the state and the highest incidence of positive cases. Cases were organized by five primary production types: broiler breeders, commercial broilers, layer breeders, commercial layers, and commercial turkeys. The greatest numbers of positive cases submitted to PADLS were associated with commercial turkey flocks (42%), followed by commercial layer flocks (27%), broiler breeder flocks (17%), commercial broiler flocks (13%), layer breeder flocks (3%), and then other/uncategorized poultry flocks (2%). In total, 118 aMPV-positive necropsy cases were assessed for prevalence of co-infections with other agents. An average of 2.4 co-infections were found per aMPV-positive necropsy case. In all production types, Escherichia coli was the most common co-infection. In chickens, Avibacterium paragallinarum, Gallibacterium anatis, and infectious bronchitis virus were other common co-infections. In commercial turkeys, Ornithobacterium rhinotracheale was a common co-infection. A retrospective analysis of aMPV serology was conducted, and 90% of serum samples from commercial layers, 85% from broiler breeders, 79% from commercial turkeys, 57% from commercial broilers, and 100% of serum samples from layer breeders were positive for aMPV antibodies assessed via commercial ELISA kits. Titer values varied based on production type, with older birds/longer-lived production types producing the highest titer values and greater proportions of positive samples. The data collected from this retrospective study provide opportunities for future analysis of the first year of infection, particularly regarding titer values of naïve vs. vaccinated or previously exposed birds, as well as potential sources/reservoirs of infection, including wild birds.

The influenza virus neuraminidase (NA) is a promising target for next-generation influenza vaccines but standardised protocols for NA-based serological assays are lacking. Previous studies have demonstrated discordant results from haemagglutination inhibition and live virus microneutralization assays when comparing matched serum and plasma samples. It is therefore important to consider the choice of serum or plasma samples in assays measuring influenza virus NA-specific antibodies. Here, we compared antibody titres against influenza A and B virus NAs in matched serum and different types of plasma using an enzyme-linked lectin assay (ELLA) and an enzyme-linked immunosorbent assay (ELISA). We observed good correlations between titres determined in serum and different types of plasma. However, there was variable and often poor agreement in the nominal titre values obtained from serum and different kinds of plasma in both ELLA and ELISA, with plasma samples often resulting in lower titres compared to serum samples. We also found differences in NA-specific responses to seasonal influenza vaccination assessed in serum versus plasma. Overall, our data suggest discrepancies between NA-specific antibody measurements in serum and plasma. Therefore, the consistent use of serum should be considered in standardising NA-based serological assays.

The production of lactic acid, a crucial platform chemical by microbial fermentation, is currently hindered by its low production efficiency. Herein, this study aims to enhance the synthetic efficiency of lactic acid in widely used Komagataella phaffii (K. phaffii) by reconfiguring the synthetic pathway and a metabolite damage-repair system. First, through the introduction of lactic acid dehydrogenase (BsLDH) from Bacillus subtilis and the transporter LutP from Bacillus coagulans, the titer of lactic acid was increased from 1.5 g/L in the original strain to 55.4 g/L using glucose. In particular, the titer was improved to 87.2 g/L by introducing metabolite damage-repair genes for NAD(P)HX detoxification and phosphate-based inhibitor elimination. In addition, the carbon source transport and metabolism pathway were strengthened, resulting in titers of 18.7 and 7.7 g/L from glycerol and methanol. Finally, the strains were scaled up in a 5 L bioreactor, achieving lactic acid titer values of 153.0, 133.2, and 37.4 g/L from glucose, glycerol, and methanol, respectively. This study significantly improved the yield of lactic acid production from low-cost carbon sources by microbial fermentation, demonstrating the potential of engineered K. phaffii for industrial production.

Infectious bronchitis virus (IBV) is an avian coronavirus and a primary causative agent of respiratory disease in poultry, representing a significant global economic concern. IBV shares structural and functional similarities with other coronaviruses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), making it an invaluable model for studying coronavirus behavior and exploring potential therapeutic strategies. Novel applications could prove valuable in controlling the spread of these pathogens. This study evaluates the antiviral efficacy of various commercial formulations of the probiotic-based disinfectant (Prowill), Astragalus root extract, and ivermectin against the IBV D274 strain (Genotype 1-Lineage 12; GI-12). The agents were applied at different concentrations to specific pathogen-free (SPF) embryonated chicken eggs (9- 10 days old) previously inoculated with IBV D274. The key parameters, such as egg mortality, mean hemagglutination (HA) titer, and HA titer (log2) values, were assessed to determine the antiviral efficacy of each treatment. The results revealed that the gel form of Prowill demonstrated superior antiviral activity, including at lower tested concentrations. This gel form showed the lowest egg mortality rates (10% at 0.5%, 1%, and 2% concentrations). This form also exhibited relatively low HA titers (4.7, 3.8, and 3.6 in log2, respectively). In addition, other Prowill formulations demonstrated significant antiviral effects, reducing mortality rates. Astragalus root extract, particularly at a concentration of 5 mg/kg, demonstrated potent antiviral activity by effectively inhibiting viral replication; however, mortality reached 30%. Ivermectin showed remarkably high cytotoxicity. These findings suggest that Prowill disinfectants, especially in their gel form, might represent a promising auxiliary application for managing the spread of IBV and potentially other coronaviruses. Further investigation is required to evaluate the long-term safety and broader applicability of these disinfectants in controlling avian coronaviruses and related pathogens.

Detection of antibodies against infectious bronchitis virus strain QX (GI-19) by commercial ELISA kits and virus neutralization test.

Porphyrins are essential tetrapyrroles that play critical roles in biological electron-transfer and light-harvesting systems. As the universal precursor of heme and chlorophyll, protoporphyrin IX (PP IX) has transformative potential for fields as diverse as biomedicine, materials, food, and agriculture. However, large-scale microbial PP IX production is subject to challenges regarding cellular toxicity and regulation of tetrapyrrole biosynthesis. Herein, we report a synthetic-biology-driven Escherichia coli platform enabled by spatially resolved pathway optimization and cytotoxicity mitigation. By introducing a hyperactive 5-aminolevulinic acid synthase and rebalancing branch pathways via sRNA-based knockdown, we decoupled the PP IX synthesis from endogenous regulatory constraints. Integration of the MacAB-TolC efflux system reduced intracellular PP IX accumulation by 16%, synergistically enhancing extracellular productivity. PP IX titer values of 3.90 g/L and 65.0 mg/L/h productivity were achieved in a 5 L bioreactor, the highest ever reported. The engineered chassis exhibits metabolic plasticity, coproducing 0.24 g/L heme through dynamic pathway modifications. This work establishes a new paradigm for cytotoxic metabolite synthesis through spatiotemporal pathway governance, circumventing classical toxicity-productivity trade-offs. Our work establishes an efficient platform for microbial PP IX production. Furthermore, the engineered chassis developed here enables versatile applications in next-generation porphyrin biomanufacturing.

Due to the limitations in antiviral treatments for viral infections, the search for natural compounds with biocompatible and antiviral activities has gained importance. In this study, we investigated the antiviral efficacy of a unique formulation (DEO/TA-mix, Uluvir®) at the stages of viral replication, adsorption, penetration, repeated doses, and direct inactivation of the selected model virus, Bovine coronavirus (BCoV). In the presence of DEO (from Anethum graveolens L.)/ TA (Quercus infectoria extract) mix, 99.94% inhibition was observed in the mean viral titer values of BCoV at the 48th h of replication, while the inhibition activity stopped at the 96th h. With the addition of DEO/TA-mix every 48h after virus inoculation, viral replication was inhibited by 98.79% at the 120th h. Treatment of BCoV with DEO/TA-mix showed 99.58% inhibition at the adsorption stage and 43.77% inhibition at the penetration stage in the viral titer. In the direct inactivation efficacy of DEO/TA-mix on BCoV, the mean viral titers decreased by 0.5 to 3.0 log in a time-dependent manner. The antiviral activity of DEO/TA-mix is predicted to be more effective in the early stages of BCoV replication. In addition, an additional dose of DEO/TA-mix every 48h during the viral replication phase increases and prolongs the inhibition rates on viral titers. This study has demonstrated that DEO/TA-mix shares high antiviral activity and may be evaluated as a potential drug for virus infections.

Validation and evaluation of the rapid fluorescent focus inhibition test (RFFIT) for the quantification of neutralizing antibodies as an indirect measure of rabies vaccine relative potency: An alternative to the NIH test.

The versatile non-biodegradable material plastic has significantly enhanced innovation, but its production heavily relies on fossil fuels and non-renewable resources, which causes severe pollution and ecosystem disruption, highlighting the urgent need for eco-friendly alternatives. Polyhydroxyalkanoates (PHA) are a class of bioplastics that offer a promising solution as biodegradable, environmentally compatible, and versatile biopolymer synthesized by microorganisms using various substrates, aiding in organic waste management. This study was aimed to isolate and profile PHA producing bacteria from diverse sources such as soil, compost, landfill site, and sewage in Nepal. The initial screening of bacteria was performed by Sudan Black B dye, followed by secondary screening with the more specific Nile Blue A dye to detect PHA accumulation. Out of 343 isolates, 81 were confirmed positive for PHA production which were further processed for PHA extraction. The isolates Ht3d (12.76 ± 1.854%), Nk3e (22.748 ± 3.608%), Mn7d (14.24 ± 2.223%), and Dg5c (14.952 ± 3.401%) from soil, compost, landfill, and sewage respectively, showed the highest PHA accumulation and were biochemically identified as Bacillus circulans, Bacillus subtilis, Staphylococcus aureus, Staphylococcus spp., respectively. The yield ratio for isolate Ht3d significantly increased by 2.74 fold under the optimal conditions (pH 7, 35 °C, and 48 h with glucose as carbon source) which was 34.99 ± 5.61% having titer value of 0.82 g/L and production rate of 0.034 g/L/h with respect to 2.4 g/L of cell biomass. The production rate and dry cell weight of other isolates were also enhanced under different optimal conditions. The characterization of the produced biopolymer through UV-Visible spectrophotometry provided maximum absorbance at 232 nm and the FT-IR spectroscopy indicated the presence of O-H and C-H vibrations along with C = O, C-O-C stretching which collectively confirmed the presence of PHA.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12866-025-04314-8.

Objective: This study aimed to evaluate and compare hepatitis B's knowledge, attitudes, and practices among dental and medical students in Iraqi universities. Material and Methods: A cross-sectional observational study was conducted from 1 July to 1 September 2024 among second, third, fourth, and fifth-stage medical and dental students. For survey distribution, the questionnaire consisting of four domains submitted to an internet platform (Google Forms). Undergraduate students were invited to participate in the research via a survey link published on specific groups of students in the social networking platforms Viber and WhatsApp. Results: 426 students responded to the questionnaire. There were statistically significant differences in frequency were seen (p < 0.001) regarding source of your knowledge about HBV, route of infection transition, receiving vaccination, doses of vaccine, and periodic checking of titer values of the vaccine. Frequency of answers for questions regarding immunization programs and strategies against HB infection were not statistically different. Additionally, dental students had more frequent positive responses for use of gloves, changing gloves, use of face mask, changing face masks, use of disposable gown, and use of disposable instruments for the patients positive or suspected HB virus (95.4%, 94.6%, 93.8%, 73.0%, 74.3%, and 96.3%, respectively) whereas medical students less frequently used those (67.6%, 62.7%, 59.5%, 24.9%, 47.6%, and 85.9%, respectively). Conclusion: The study revealed that although both medical and dental students were knowledgeable about HBV transmission, vaccination uptake, particularly among medical students, was inadequate.

Protective efficacy of Interferon-γ and β-glucan adjuvanted formalin killed vaccines in Nile tilapia against Edwardsiella tarda infection.

Background: Columnaris, Aeromonas infestations and Vibriosis, are considered highly prevalent bacterial infections. A. salmonicida is considered a facultatively anaerobic Gram-negative bacterium. The valuation of conservation status of Emerita has specifically challenging owed to inherent complexities in finding its peaks and reproductive cycles which need the prior and the extensive knowledge and also monitors the gsssenerations and populations in life cycle i.e. pelagic and benthic, and the diversity of the stressors and the anthropogenic influences on marine environments and beaches, where creating the distinctions within the effects and causes is complex. Aim: The present study examines the mole crab's immune reaction and determines the hemagglutinin value of the Emerita asiatica. Methodology: For serum separation and microbe agglutinating activity, hemagglutination and cross adsorption experiments have been conducted in this study. All of the investigated bacterial species responded with the agglutinin that has extracted from the infected microorganisms. The mole crab's defensive system against bacterial infection has been shown by bacterial inhibition and bacterial agglutination. Various RBCs are used in Hemagglutination. Various Bacteria are used in Bacterial Agglutination and Inhibition. Yeast and trypsinized yeast are used in Yeast Agglutination. Results: When tested with Buffalo RBC, tripsinized yeast, and Bacillus subtilis, the serum produced the highest HA titer. The trypsinized yeast has a higher titer value for agglutination compared to the native yeast. It was discovered that the hemagglutinating activity was strong between temperatures of 20 and 50 ̊C. Conclusion: Based on its physicochemical study, HA is thermally stable, pH-stable (between 7 and 9), and activity-dependent on the presence of Ca2+. The HA titers of serum samples tested against buffalo RBC were reduced after extensive dialysis in a divalent cation-free buffer. Serum calcium levels in mole crabs ranged from 0.9 to 1.9 mg/ml. Serum of Emerita asiatica with erythrocytes had the highest HA activity. Emerita asiatica serum hemagglutinating activity was unaffected by biochemical factors such water, protein, and calcium content.

Vaccindo is the first and most trusted local vaccine producer to meet the vaccine needs of farmers in Indonesia. This study was to determine the quality of IBH who is still in the trial phase, vaccine production at PT. Vaksindo and to determine the application of defect control with Six Sigma method on defective vaccine products. to obtain the concentration of the virus produced on average does not enter the minimum threshold of the standard set, that is 7.0 TCID50 /0.1 mL in trial phase, Defect Per Million Opportunity (DPMO) is 826408.73, which means that in every million times of product produced in each production has a defective product of 826408.73, if it is converted into a sigma table the DPMO value does not even enter the first sigma level is the value of 690,000 DPMO, which means the process carried out in the IBH vaccine production process by PT. Vaksindo coverage is still below 30.9% to improve this, improvements are such as testing the level of viruses of VCT titration in each sample post harvest and pre production to maintain the quality of the antigen produced, as well as propagation seed is done at the maximum limit that can be obtained so that when propagated to antigen, the titer value or concentration of the virus does not go down and remaining standard.

Objective — to determine the relationship between the characteristics of the hormonal background and signs of FSD in women with ATD in the prediction of the success of future gestation and the formation of POF. Materials and methods. The study design is a prospective study that included 30 women of reproductive age (main group) with ATD and 30 healthy women of reproductive age — the control group. The levels of steroid and peptide hormones, domains of FSD and the ovulatory phase of the menstrual cycle were studied during 9 months of therapy. The presence of female sexual dysfunction was determined using a questionnaire with scoring on an online calculator. The treatment included the use of a complex of herbal remedies recommended for hypoandrogenic ovarian dysfunction. Statistical analysis was performed using the specialized statistical software package SPSS 25.0 (StatSoft Inc., USA). Results. All women in the main group had a combination of signs of FSD with anovulation against the background of clinically significant TPOAb indicators. Correlations were established: statistically significant relationship between desire and 17-OP indicators; arousal and progesterone; orgasm and TPOAb, TSH, progesterone; pain and homocysteine, TSH; FSFI and TSH, between questionnaire scores and hormonal indicators: orgasm and vitamin D; AMH and pleasure, pain, FSFI (p<0.05). After 9 months of therapy, 60.0% (n=18) of the patients in the main group had persistent DHEA-S deficiency. Monitoring of ovulation by ultrasound folliculometry in the main group indicated a negative prognosis for natural pregnancy in 16 (53.3%) of the examined. Conclusions. Familiarity of doctors of related specialties with the problem of autoimmune thyroid pathology and its consequences for the sexual and reproductive health of women can improve their awareness of the individualized prognosis of pregnancy. Pregnancy planning in women with ATD with an emphasis on their age and the value of thyroid antibody titers. Determination of reproductive steroidogenesis along with examination of the thyroid panel. With low DHEA-S indicators — indications for examination of adrenal function.

Succinic acid (SA) necessitates thorough examination regarding its biological production pathway. The energy-dependent biosynthesis of SA and the resulting intracellular redox imbalance contribute to reduced SA productivity. Co-culture fermentation that combines aerobic yeast and facultative bacteria demonstrates a dual benefit for microbial growth and SA production. The optimization of the fermentation process with native SA producers resulted in higher SA titer values. The intermittent addition of bicarbonate salts increased SA production to 3.86g L⁻¹ h⁻¹ during the anaerobic fermentation of Actinobacillus succinogenes. Fed-batch fermentation of acetic acid enhanced SA production to 12g/L via a glyoxylate shunt in Yarrowia lipolytica. The immobilization of SA-producing microbial strains enhanced continuous fermentation, resulting in SA titers and productivity of 69g/L and 35.6g L⁻¹ h⁻¹, respectively. Metabolic enhancements of SA yield are increasingly documented through various rational engineering approaches applied to bacterial and yeast strains. This review paper aims to analyze the challenges associated with conventional SA fermentation processes. This work examines literature on SA production from lignocellulosic biomass, offering a comprehensive analysis of the valorization of complex biological resources. This paper emphasizes recent advancements in SA yields, reaching nearly 0.98g SA/g substrate, attained through innovative methodologies.

Monoclonal antibodies (mAbs) targeting the SARS-CoV-2 receptor-binding domain are used to treat and prevent COVID-19. However, the rapid evolution of SARS-CoV-2 drives continuous escape from therapeutic mAbs. Therefore, the ability to identify broadly neutralizing antibodies (bnAbs) to future variants is needed. Here we use deep mutational scanning to predict viral receptor-binding domain evolution and to select for mAbs neutralizing both existing and prospective variants. A retrospective analysis of 1,103 SARS-CoV-2 wild-type-elicited mAbs shows that this method can increase the probability of identifying effective bnAbs to the XBB.1.5 strain from 1% to 40% in an early pandemic set-up. Among these bnAbs, BD55-1205 showed potent activity to all tested variants. Cryogenic electron microscopy structural analyses revealed the receptor mimicry of BD55-1205, explaining its broad reactivity. Delivery of mRNA-lipid nanoparticles encoding BD55-1205-IgG in mice resulted in serum half-maximal neutralizing antibody titre values of ~5,000 to XBB.1.5, HK.3.1 and JN.1 variants. Combining bnAb identification using viral evolution prediction with the versatility of mRNA delivery technology can enable rapid development of next-generation antibody-based countermeasures against SARS-CoV-2 and potentially other pathogens with pandemic potential.

Nile tilapia (Oreochromis niloticus) farms face streptococcus agalactiae infections causing mass mortality following economic losses. To date, no commercial fish vaccine is available in India to protect against streptococcosis due to a lack of proper adjuvants and vaccine targets. Multiple studies justify that immunomodulators like β-glucan and Alkoxy glycerol (AKG) have improved immunity in fish. β-glucan and AKG are natural homopolymers and lipids, respectively, which have proven immunostimulant capabilities in fish and can be effectively used as a prominent adjuvant to improve vaccine efficacy. The present study aims to demonstrate the improved adjuvant efficacy of the formalin-inactivated S. agalactiae vaccine. Nile tilapia fingerlings were immunised with formalin-inactivated S. agalactiae vaccines mixed with β-glucan and AKG. Each week postvaccination, the serum nonspecific responses were measured by lysozyme, myeloperoxidase, SOD and bactericidal activity. Following the challenge after 28 days postvaccination, the survival rate achieved was 32.14%, 57.14%, 53.57% and 71.43% in the respective experimental groups. The immune gene expressions of IgM, TCRβ and MHCII were evoked postvaccination. The specific serum IgM titre values were significantly upregulated in 14- and 28-day postvaccinated experimental groups. The study highlights the enhanced activity of β-glucan and AKG incorporated vaccine, which has the potency towards commercialisation for sustainable aquaculture.

Changes in serotype or genetic payload of recombinant adeno associated virus (rAAVs) gene therapies require adapting the transfection conditions of the upstream HEK293 cultivations. This study adopts an iterative model-based experiment design approach, where increasing data availability is leveraged to evolve models of different complexity. Initial models based on data from shaker flask runs guided the design of the first round at Ambr250 scale. With Ambr250 data becoming available, hybrid models capturing process state evolutions and historical models incorporating these evolutions to predict rAAV titer, were developed. These models were then combined into a full model approach, which was utilized within a Bayesian Optimization framework for the design of a second round of Ambr250 scale runs. The iterative approach was tested across different projects applying transfer learning to enhance the predictive power and improve the subsequent optimization. The approach was benchmarked against a statistical Design of Experiment method. The results show that the model-based experiment design consistently (and across projects) produces higher rAAV titer values than the benchmark approach (Project C: 4.4% or 7.0% increases in titer values relative to the response surface modeling approach for ELISA and ddPCR, respectively; Project D: 32.4% or 10.9% increases in titer values relative to the standard DoE-screening pick for ELISA and ddPCR, respectively), effectively optimizing the transfection mixture composition. The combination of propagation and historical models, augmented by transfer learning and an ever-increasing amount of data, enhanced the process design workflow, contributing to improved rAAV production through efficient transfection strategies.

The diagnosis of typhoid fever can be made by hematological examination and the Widal method. Hematologic change that often occurs in typhoid fever is thrombocytopenia. Platelet function can be observed by examining the platelet index, namely the Mean Platelet Volume (MPV), and Platelet Distribution Width (PDW). The titer on the serological examination of the Widal test is a significant diagnostic for infections caused by the bacterium Salmonella typhi. Objectives: To determine the relationship between the Widal titer and the number and index of platelets in patients with typhoid fever at the West Lombok Health Center. Methods: The research methodology used is an analytic observational method with a cross-sectional approach. This study was conducted from February 2021 to April 2021. The population in this study was all patients who underwent laboratory examinations diagnosed as positive for typhoid fever in the Gunungsari Health Center, West Lombok Regency, with a sample of 24 respondents. Data analysis using Spearman correlation. Results: Statistical test results obtained P≥0.05, namely the titer value against platelets p=0.429, the titer against the MPV platelet index p=1,000, PDW p=0.291. Conclusion: there is no relationship between widal titer to the number of platelets and the platelet index in patients with typhoid fever.Keywords:  typhoid fever, platelets, platelet  index, widal titer

Rabies is caused by a virus from the genus Lyssavirus, which belongs to the Rhabdovirus family. In Badung Regency, 13 cases of rabies in dogs were confirmed in 2015, with 8 positive cases reported in Mengwi District. This study aims to measure the antibody titer against the rabies virus in local dogs that had been vaccinated in Penarungan Village, Mengwi District, Badung Regency, seven months after vaccination. The method used in this study was antibody titer testing through the Enzyme-linked Immunosorbent Assay (ELISA). Samples were selected based on inclusion criteria and were collected from Banjar Blungbang, Banjar Cemenggon, Banjar Guming, Banjar Umopoh, and Banjar Bantas. A total of 16 blood samples from local dogs were collected using the venipuncture technique. The results of the study using the ELISA test showed that 12 samples were seropositive, with an average titer value of 0.9 IU/mL, while 4 samples were seronegative, with an average titer value of 0.4 IU/mL. The percentage of seropositive antibody titers in local dogs in Penarungan Village seven months after rabies vaccination was 75%, while the percentage of seronegative antibody titers was 25%. Therefore, booster rabies vaccinations are necessary for dogs with seronegative antibody titers.