Specific IgM Antibodies Research Articles

Effect of the MONTANIDE™ IMS 1312 adjuvant on the innate and adaptive immune responses of Nile tilapia (Oreochromis niloticus) against Streptococcusagalactiae through immersion vaccination

Streptococcus agalactiae is a common pathogenic bacterium caused of streptococcosis, which has a negative impact on Nile tilapia aquaculture. Numerous vaccines have been developed recently to combat this disease, which are key components of global health efforts to prevent disease outbreaks. MONTANIDE™ IMS 1312 is a micro-emulsion recommended for immersion of fish. However, the data on the effectiveness of those immersion vaccines containing this aqueous adjuvant in fish are limited. The objective of this research was to explore the potential of MONTANIDE™ IMS 1312, an adjuvant for immersion vaccination, administered with an S. agalactiae inactivated whole-cell vaccine (SAIV) in Nile tilapia. Fish were separated into three groups: 1) fish were vaccinated by immersion vaccination with PBS (CTRL), 2) fish were vaccinated by immersion vaccination with SAIV vaccine alone (SAIV), and 3) fish were vaccinated by immersion vaccination with SAIV containing MONTANIDE™ IMS 1312. We found that the activity of several innate immunity parameters was increased significantly (P < 0.05) following the immunization. As expected, the levels of specific IgM antibody were significantly increased post-vaccination, and the highest IgM antibody levels were found in the fish vaccinated with SAIV containing MONTANIDE™ IMS 1312. Analysis of the transcriptional expression of major pro-inflammatory cytokines, as well as the presence of IgM+ B cells, revealed significant increases, suggesting that Nile tilapia were able to initiate cellular immune responses following vaccination. Taken together, our results indicate that using MONTANIDE™ IMS 1312 in combination with a SAIV can induce strong protection post S. agalactiae infection. Importantly, administration of an adjuvanted immersion vaccine is safe, no side effects were observed, and it does not negatively impact fish growth. In conclusion, MONTANIDE™ IMS 1312 has the potential to be used as adjuvants for immersion vaccines against streptococcosis in Nile tilapia.

A novel multiplex and glycoprotein-based immunochromatographic serologic IgM test for the rapid diagnosis of Escherichia coli O157 and O145 causing bloody diarrhea and hemolytic uremic syndrome.

Shiga toxin-producing Escherichia coli (STEC) are the main etiological agents of hemolytic uremic syndrome (HUS). Good clinical management of STEC infections and HUS depends on early, rapid, and accurate diagnosis. Here, we have developed and evaluated the first multiplex and glycoprotein-based immunochromatographic test for the detection of IgM antibodies against the O-polysaccharide of the lipopolysaccharide of E. coli O157 and O145 in human serum samples. A retrospective study was carried out resulting in a diagnostic sensitivity of the E. coli O157/O145 LFIA (lateral flow immunoassay) of 97.1% and 98.9% for O157 and O145, respectively, and 97.9% for both serogroups. The diagnostic specificity was 98.7% for O157 and O145, and the overall specificity 97.4%. In samples obtained before 3 days after the onset of diarrhea, the detection percentage was 83%, increasing to 100% from 3 days onward. Finally, the association of bloody diarrhea (BD) or HUS cases to an STEC infection increased from 22.8% to 77.2% when stool culture and stx/Stx detection were combined with serology by LFIA. Our results demonstrate that the E. coli O157/O145 LFIA is a highly accurate and serospecific test for the early and rapid diagnosis of E. coli O157 and O145 infections in BD or HUS cases. This test allows the detection of specific IgM antibodies very early in the course of the infection, making it an ideal diagnostic tool to be implemented in pediatric emergencies and, thus, avoid delays in the application of the correct supportive or specific treatment and prevent complications associated with HUS.

Dengue fever as autochthonous infectious disease in Italy: Epidemiological, clinical and virological characteristics

BackgroundSince August to November 2023, 82 cases of autochthonous or non-travel related Dengue virus (DENV) infection have been reported in Italy, highlighting a concerning trend of local transmission. We describe the clinical and laboratory findings of 10 autochthonous DENV in the metropolitan area of Rome admitted to the Lazzaro Spallanzani National Institute for Infectious Diseases. Method and resultsTen patients (3 males, 7 females; median age: 51) with classic dengue fever symptoms were admitted between August and November 2023. Laboratory tests confirmed dengue infection through DENV non-structural protein 1 and/or immunoglobulins (IgM/IgG) positive tests, moreover leukopenia, thrombocytopenia, elevated transaminases were detected. A subset of patients underwent extensive biological sampling, including real-time RT-PCR and immunofluorescence, to monitor DENV-RNA and antibody levels over 30 days. DENV-1 was detected in 8 patients and DENV-3 in 2. Upon admission specific IgM antibodies were found in 7 patients while IgG antibodies in 4 patients. DENV RNA was consistently detected in blood within the first 8 days but was less common in saliva and urine. No DENV RNA was detected after day 24. ConclusionThese findings contribute to the understanding of the clinical course of DENV infection in a non-endemic setting as integrated epidemiological and clinical model to increase syndromic surveillance and timely diagnosis of DENV infections.

The role of aroA and ppk1 in Aeromonas veronii pathogenicity and the efficacy evaluation of mutant strain AV-ΔaroA/ppk1 as a live attenuated vaccine

Aeromonas veronii is an opportunistic pathogen that poses great threat to aquaculture and human health, so there is an urgent need for green and efficient methods to deal with its infection. In this study, single and double gene deletion strains (AV-ΔaroA, AV-Δppk1 and AV-ΔaroA/ppk1) that can be stably inherited were constructed. Pathogenicity test showed that the toxicity of AV-ΔaroA and AV-ΔaroA/ppk1 was significantly lower compared to wild-type A. veronii. Biological characterization analysis revealed that the decrease in pathogenicity might be due to the declined growth, motility, biofilm formation abilities and the expression of virulence-related genes in mutants. Subsequently, we evaluated the efficacy of AV-ΔaroA/ppk1 as a live attenuated vaccine (LAV). Safety assessment experiments showed that AV-ΔaroA/ppk1 injected at a concentration of 3 × 107 CFU/mL was safe for C. carassius. The relative percentage survival of AV-ΔaroA/ppk1 was 67.85 %, significantly higher than that of the inactivated A. veronii, which had an RPS of 54.84 %. This improved protective effect was mainly attributed to the increased levels of A. veronii specific IgM antibody, enhanced alkaline phosphatase, lysozyme and superoxide dismutase activities, as well as higher expression levels of several immune related genes. Together, these findings deepen our understanding of the functional roles of aroA and ppk1 in A. veronii pathogenicity, provide a good candidate of LAV for A. veronii.

No detection of tick-borne encephalitis virus RNA in blood, urine or saliva of hospitalised immunocompetent tick-borne encephalitis patients.

Tick-borne encephalitis (TBE) is usually diagnosed based on the presence of TBE virus (TBEV)-specific IgM and IgG antibodies in serum. However, antibodies induced by vaccination or cross-reactivity to previous flavivirus infections may result in false positive TBEV serology. Detection of TBEV RNA may be an alternative diagnostic approach to detect viral presence and circumvent the diagnostic difficulties present when using serology. Viral RNA in blood is commonly detectable only in the first viremic phase usually lasting up to two weeks, and not in the second neurologic phase, when the patients contact the health care system and undergo diagnostic work-up. TBEV RNA has previously been detected in urine in a few retrospective TBE cases in the neurologic phase, and furthermore RNA of other flaviviruses has been detected in patient saliva. In this study, blood, saliva and urine were collected from 31 hospitalised immunocompetent patients with pleocytosis and symptoms of aseptic meningitis and/or encephalitis, suspected to have TBE. We wanted to pursue if molecular testing of TBEV RNA in these patient materials may be useful in the diagnostics. Eleven of the 31 study patients were diagnosed with TBE based on ELISA detection of TBEV specific IgG and IgM antibodies. None of the study patients had TBEV RNA detectable in any of the collected patient material.

The Prevalence of Toxoplasma Gondii Infection in Duhok, Kurdistan Region, Iraq

Toxoplasmosis is a parasitic infection, with widespread gestational and congenital transmission rates globally, consequently, it is regarded as a public health concern and an overlooked medical condition. Human infection occurs through various routes, including the ingestion of oocysts found in cat feces, consumption of raw or undercooked meat containing the parasite, and vertical transmission from an infected pregnant mother to her fetus. Objective. The aim of this study is to determine the prevalence of Toxoplasma gondii infection among individuals in various age groups and genders in Duhok City. Methods. In this research, 2251 individuals of various ages and genders visited the central laboratory in Duhok Сity, Kurdistan Region, Iraq, and peripheral blood samples were taken from individuals to identify specific anti-Toxoplasma gondii IgM and IgG antibodies using serological test (ELISA test). Results. Based on the findings of this study, it was observed that the prevalence of toxoplasmosis among females with infections detected in both IgM and IgG was 0.3 % and 20.6 %, respectively, and in males, the rate of infection with IgG was found to be 0.3 %, while no infections were detected with IgM. Also, among female individuals, the study recorded a 0.3 % infection rate for toxoplasmosis, which encompassed both IgM and IgG cases. Conclusions. Based on this study, it can be concluded that Toxoplasma infection is more common in females than in males, chronic toxoplasmosis appears to be more prevalent than acute infection with the highest rate of acute infections was found in the age group of 21–40 years.

Diagnostic Significance Vasoendothelial Growth Factor Depending on the Severity and Duration of Atopic Dermatitis, as Well as Taking into Account the Presence of Markers of Herpes Virus Infection

Currently, the problem of diagnosis and treatment of diseases associated with disorders of the angiogenesis process, as well as regenerative processes, is acute. Factors regulating the processes of angiogenesis in allergic diseases, including atopic dermatitis, play a key role in maintaining chronic inflammation and can have a significant impact on the course of the disease. Materials and methods: The study is analytical cross-sectional and presented by a comprehensive examination of 140 patients with AtD aged 2 to 12 years (median age 4.2 years), divided into 2 groups: 70 children with an established diagnosis of AtD; 70 children with atopic dermatitis infected with herpes simplex virus (AtD+HVI). The control group consisted of 70 somatically healthy children. A special laboratory examination included the determination of specific IgM and/or IgG class antibodies to herpes simplex virus type 1-2 antigens by enzyme immunoassay (ELISA); determination of the DNA of the studied herpesviruses in blood samples by polymerase chain reaction; determination of vascular endothelial growth factor A (VEGF-A) in the blood plasma of patients by ELISA. The results of our own research: A statistically significant (p &lt;0.001) increase in the level of vascular endothelial growth factor A in blood serum was found in children with AtD compared with the control group. Against the background of infection with the herpes simplex virus, an increase in the level of vascular endothelial growth factor A in blood serum was revealed compared with patients with atopic dermatitis (p &lt; 0.001). There was also a statistically significant increase in serum VEGF-A levels in patients with AtD (p &lt; 0.001) and AtD+HVI (p &lt; 0.001) with an increase in the severity of AtD. This was confirmed by the results of a correlation analysis that revealed the relationship between the level of VEGF-A and the severity of clinical symptoms of the disease.The addition of herpesvirus infection to AtD worsens the clinical symptoms of this disease.

Laboratory diagnosis and epidemiological features of leptospirosis cases in a tribal region of Western Maharashtra, India: A retrospective study

PurposePalghar district, located in the coastal region of the Konkan division of Maharashtra, has a predominantly tribal population. Leptospirosis is a major neglected public health problem and is highly underreported in Palghar district. The study aimed to evaluate the seropositivity of Leptospira infection and its associated epidemiological factors in tribal areas of the Palghar district of Maharashtra. MethodsThe present retrospective study included 94 samples of patients clinically suspected of leptospirosis during a period of one year (2021–2022) tested at Model Rural Health Research Unit (MRHRU) Dahanu. The serum sample testing was done for the presence of specific Leptospira IgM antibodies using the Panbio™ Leptospira IgM ELISA kit. Leptospirosis seropositivity was correlated with various epidemiological risk factors. ResultsA total of 12 samples of patients tested positive for specific IgM antibodies by ELISA method, indicating an overall positivity of 12.8%. Among those who tested positive, fever (83.3%), headache (58.3%), myalgia (50%), redness of the eyes (50%), and calf tenderness (16.7%) were the common symptoms observed. Subjects with redness of the eyes were significantly associated with leptospirosis (p = 0.018). The highest positivity (50%) was reported from the Ganjad area of Dahanu taluka. Farmers and animal handlers were most affected by leptospirosis. ConclusionThe high proportion of Leptospirosis cases reflects the endemic nature of the disease in the Palghar district. This study shows seasonal trends in leptospirosis incidence over the year. The clinical presentation of leptospirosis may vary from sub-clinical to mild illness to severe and potentially fatal. The findings of this study will be important for achieving the overarching goal of One Health.

West Nile virus meningoencephalitis and cytokines analysis: review and case report

West Nile virus (WNV) is transmitted via mosquito bites and causes ubiquitous zoonosis. Most infections in humans are asymptomatic, approximately 20% present as fever, and less than 1% as neuroinvasive disease. Central nervous system involvement is presented as meningitis, encephalitis, acute flaccid paralysis, or a combination of them. West Nile neuroinvasive disease (WNND) has a severe clinical course, potentially fatal outcome and frequent neurological sequelae in survivors. Risk factors for neurological impairment are advanced age and immunosuppression. Here we present a clinical case of meningoencephalitis caused by WNV and a brief literature review. Clinical and epidemiological data, laboratory, microbiological, molecular methods and imaging techniques were used. The cytokine levels of IL-6, IL-8, IL-10, IL-12(p40) and TNF-α in cerebrospinal fluid (CSF) and serum were also measured. We present a 55-year-old man with a sudden onset of headache, vomiting and fever. The symptoms appeared after a recent trip to Türkiye and involved multiple mosquito bites. Neck stiffness, disturbances in consciousness, signs of cerebral edema and subsequently focal neurological deficits were observed. The etiological diagnosis was verified by positive polymerase chain reaction for WNV and the presence of specific IgM antibodies in the CSF. After a 28-day hospital stay, the patient was discharged and referred to a Physiotherapy Unit due to residual motor deficits. WNV etiology should be suspected in patients with clinical and laboratory signs of viral neuroinfection, mosquito bites, and/or travel to endemic regions.

Clinical and historical infection of Tacheng tick virus 2: A retrospective investigation.

Tacheng tick virus 2 (TcTV-2) is an emerging tick-borne virus belonging to the genus Uukuvirus in the family Phenuiviridae. Initially isolated in 2019 from a patient in Xinjiang Uygur Autonomous Region (XUAR), northwestern China, who developed fever and headache after a tick bite, TcTV-2 was concurrently molecularly detected in hard ticks across various countries, including China, Kazakhstan, Romania, and Turkey. This study conducted a retrospective epidemiological investigation of TcTV-2 infection. In this retrospective cohort study, we collected samples from 47 tick-bitten patients, 984 herdsmen, 7 Asian badgers, 13 red foxes, and 168 Hyalomma asiaticum tick egg batches. Patients' samples were primarily analyzed by using high-throughput sequencing, targeting the V3-V4 region of the bacterial 16S rRNA gene and viral cDNA libraries. Typical tick-borne pathogens were further confirmed using RT-PCR and detected in Asian badgers, red foxes and Hy. asiaticum tick egg batches. We also conducted enzyme-linked immunosorbent assay (ELISA) to detected specific IgM and IgG antibodies against TcTV-2 in herdsmen. Phylogenetic analysis was performed to genetically characterize TcTV-2 detected in this study. TcTV-2 was detected in various samples, including blood, urine, and throat swabs from 12.77% (6/47) tick-bitten patients. It was found in blood samples of 14.29% (1/7) of wild badgers, 7.69% (1/13) of red foxes, and 13.69% (23/168) of Hy. asiaticum egg batches. Furthermore, ELISA results revealed that 9.55% (94/984) of the serum samples (34 from males and 60 from females) were tested positive for TcTV-2-specific IgG, while 2.95% (29/984, 7 males and 22 females) showed positivity for TcTV-2-specific IgM. Additionally, 1.02% (10/984, 4 males and 6 females) of the sera tested positive for both TcTV-2-specific IgM and IgG. Phylogenetic analysis indicated that the TcTV-2 strains detected in this study were genetically similar, regardless of their origin and host species. Clinical symptoms of TcTV-2 infection in patients are nonspecific, with common symptoms including headache, fever, asthenia, vomiting, myalgia, rash, and meningitis-like signs. TcTV-2 can be detected in blood, urine, and throat swab samples of infected patients. Among local herdsmen, 9.55% tested positive for TcTV-2-specific IgG and 2.95% for TcTV-2-specific IgM. Importantly, TcTV-2 can be transovarially transmitted in Hy. asiaticum ticks, and the Asian badgers and red foxes are potential reservoirs of TcTV-2.

Evaluating the efficiency of ELISA, monoplex and multiplex probe-based real-time reverse-transcription PCR assays in the detection of SARS-CoV-2 (COVID-19) and influenza A and B viruses: A cross-sectional study.

The current study aimed to evaluate the efficiency of Enzyme-linked immunosorbent assay (ELISA) assay and monoplex and multiplex real-time reverse-transcription PCR (rRT-PCR) in the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and influenza A and B viruses (Flu A and Flu B). The SARS-CoV-2 -specific IgG and IgM antibodies, as well as, Flu A (H1N1 and H3N2 serotypes) and Flu B virus antibodies were determined by ELISA assay. The one-step qRT-PCR method was used to detect the SARS-CoV-2 in nasopharyngeal swab samples. Furthermore, the presence of Flu A and B viruses was evaluated using probe-based RT-PCR. Simultaneous detection of SARS-CoV-2, Flu A and B viruses was performed by multiplex rRT-PCR assay. SARS CoV-2 IgM and IgG antibodies were detected in 33.3% and 58.3% of patients, respectively. In contrast, the SARS CoV-2 genome was detected in 50% of patients using the one-step monoplex RT-PCR assay. Flu A serotypes H1N1 and H3N2 were found in 16.7% and 8.3% of patients. Probe-based RT-PCR revealed that 39.3% of patients were positive for the Flu A virus. Multiplex rRT-PCR detect the SARS-CoV-2, Flu A, and Flu B in 50%, 39.3%, and 19% of samples, respectively. The sensitivity and specificity of multiplex rRT-PCR assay in comparison to monoplex RT-PCR were 100% and 55%, respectively. Coinfection with SARS-CoV-2, Flu A, and Flu B viruses was found in 9.5% of patients. Multiplex rRT-PCR can be used as a repaid, cost-effective and suitable tool for molecular surveillance of SARS-CoV-2 and Flu A/B viruses.

Immunoprotective efficacy evaluation of OmpTS subunit vaccine against Aeromonas hydrophila infection in Megalobrama amblycephala

Bacterial septicemia in freshwater fish is mainly caused by Aeromonas hydrophila infection, which affects the development of aquaculture industry. In the context of sustainable aquaculture, subunit vaccines are of great values because they play positive roles in reducing the overuse of antibiotics and protecting aquatic animals against bacterial infection. In this study, the recombinant outer membrane protein OmpTS of A. hydrophila were used as subunit vaccine to immunize Megalobrama amblycephala, and its immunoprotective effect and host immune responses were evaluated. The survival rates of the vaccinated groups after bacterial infection were significantly higher than that of the control group, especially of the OmpTS high-dose vaccinated group. The better protective effects of vaccinated groups might be attributed to the increased levels of serum IgM-specific antibody titer, the reduced relative abundance of A. hydrophila in various tissues, the increased number of immune-positive cells with different epitopes, the up-regulated expression levels of immune-related genes, and the enhanced activities of antibacterial enzymes. In conclusion, OmpTS subunit vaccine could strongly induce host immune responses in M. amblycephala, thereby enhancing both cellular and humoral immunity, which exhibited excellent and effective immunoprotective efficacy.

Serum inflammatory cytokine IL-4 and IL-18 Levels in Patients with Typhoid Fever in Baghdad

Background: Gram-negative Salmonella enterica serotype typhi, or S. typhi, is intracellular bacteria belong to Enterobacteriaceae family that infect humans and cause Salmonellosis (typhoid fever), or (enteric fever), this disease is a major public health problem and a life-threatening bacterial infection in developing countries. Every year, there are thought to be between 12 and 33 million cases of typhoid fever, which leads to over 600,000 deaths. Specific immunity both humoral and cellular specific immune response activated to control Salmonella infection. This study aimed evaluate the function of IL-4 and IL18 levels as a diagnostic aid for this infection. Material and methods: Ninety suspected patients with typhoid fever diagnosed by specialist according to sign and symptom and thirty apparently healthy control where enrolled during the study period from September 2023 to February 2024. Blood was taken from both groups (5ml) and serum were gathered using centrifugation (3000 rpm for 5 min) and kept frozen in three Eppendorf for detection of serum IgM, IgG, IL-4 and IL-18 levels. Results: the result showed that out of 90 blood samples only 60 samples were positive to salmonella infection (confirmed by detecting specific salmonella IgM and IgG antibodies using ELIZA kits Sunlong biotech\China). Serum levels of IL-4, and IL18 (pg/ml) was measured in positive samples using ELIZA kits (Cloud-clone corp \USA). There is a significant increase in the level of IgM (0.33 ± 0.13) (mg\dl) and IgG (0.27 ± 0.1) (mg\dl) as compared to the control group in patients (0.16±0.03 and 0.12±0.03) (mg\dl) respectively with P &lt; 0.01, and there was a significant increase in the average IL-4 (334.00 ± 87.49) (pg/ml) and IL-18 level in patients (380.44 ± 68.95) (pg/ml) compared to the control group (83.40 ± 17.44 and 189.66 ± 70.40) (pg/ml) respectively with P &lt; 0.001. The results indicated a direct correlation between IL-4 and IL-18 serum levels with high level of serum IgM antibody compared to their levels in control group.

Evaluation of the efficacy of MONTANIDE™ GR01, a new adjuvant for feed-based vaccines, on the immune response and protection against Streptococcus agalactiae in oral vaccinated Nile tilapia (Oreochromis niloticus) under laboratory and on-farm conditions

Streptococcosis, an emerging infectious disease caused by Streptococcus agalactiae, has had adverse effects on farmed tilapia. Several vaccines have been developed to prevent this disease and induce a specific immune response against S. agalactiae infection. In this study the use of MONTANIDE™ GR01, a new adjuvant for oral vaccination, was optimized for use in tilapia under laboratory and field studies. In the laboratory trial the immune response and protective efficacy of two doses of MONTANIDE™ GR01, 20 % (w/w) and 2 % (w/w), included into the feed-based adjuvanted vaccines were assessed comparatively. Following immunization, the innate immune parameters studied in serum, including lysozyme, myeloperoxidase, catalase and glutathione peroxidase activity, were all increased significantly. Furthermore, specific IgM antibodies against S. agalactiae were induced significantly in serum post-vaccination, with higher levels observed in both groups that received the feed-based adjuvanted vaccine. Under both injection and immersion challenge conditions, the relative percent survival for the feed-based adjuvanted vaccine groups ranged from 78 % to 84 %. Following use of the low dose concentration of MONTANIDE™ GR01 for oral vaccination of tilapia in cage culture systems, several innate immune parameters were effectively enhanced in the immunized fish. Similarly, the levels of specific IgM antibodies in the serum of feed-based vaccinated fish were significantly enhanced, reaching their highest levels 2–5 months post-vaccination. Cytokines associated with innate and adaptive immunity were also examined, and the expression levels of several genes showed significant up-regulation. This indicates that both cellular and humoral immune responses were induced by the feed-based adjuvanted vaccine. The economic impact of a feed-based adjuvanted vaccine was examined following vaccination, considering the growth performance and feed utilization of the fish. It was found that the Economic Performance Index and Economic Conversion Ratio were unaffected by vaccination, further demonstrating that there are no negative impacts associated with administering a feed-based vaccine to fish. In conclusion, the data from this study indicate that MONTANIDE™ GR01 is a highly valuable adjuvant for oral vaccination, as demonstrated by its ability to induce a strong immune response and effectively prevent streptococcal disease in Nile tilapia.

Transmission of yellow fever vaccine virus from breast feeding mothers to their infants: reporting of yellow fever virus (YFV) RNA detection in milk specimens

Background Because of yellow fever’s serious impact on health, vaccination is the principal strategy to control the disease. Administration of the yellow fever vaccine to breastfeeding women should be before they complete 9 months post-delivery, in order to prevent transmission of the yellow fever vaccine virus to their infants through breast feeding. This study aimed to confirm whether the excretion of yellow fever vaccine virus is in milk of vaccinated breastfeeding mothers and to confirm the probable transmission to their infants through breast milk. Methods Samples were taken as follows: one serum specimen was taken 3-14 days after the date of the vaccination, and breast milk specimens were taken at four different time points between 3-4 days apart. Specimens were obtained from eight nursing mothers, who received the YVF vaccine (17DD). Mothers were asymptomatic before and after the vaccine administration but their infants developed symptoms after administration. Maternal serum samples were tested for YFV specific IgM antibodies through immuno-fluorescent assay (IFA). RNA was extracted from serum and breast milk specimens and YFV RNA screened using real-time polymerase chain reaction (RT-PCR). Results In total, five mothers (62.5%) were positive for YFV and two mothers (25%) had YFV RNA in serum. Among milk specimens, YFV RNA was detected during the four different mentioned collection times as follows (positive milk specimens/total milk specimens): 3/8 (37.5 %), 4/6 (66.6%) and 1/4(25%). RNA was completely undetectable in the last collection time. Conclusions YFV transmission from mothers to their babies through breast-feeding was highly probable indicated by the temporal relationship to mother’s YF vaccination.

Diagnosis of Chronic Brucellosis with Interferon-Gamma Quantification

Background: Brucellosis can vary in severity and duration, requiring multiple tests for diagnosis. Its prevalence in Iraq complicates the interpretation of test results. Objective: This study aimed to develop a diagnostic tool leveraging variations in γ interferon to aid the immune system in recognizing illness. Methods: A comparative study was conducted at Anbar Teaching Hospital from October 2022 to September 2023. 86 blood samples were collected from the outpatient infectious clinic, comprising 43 confirmed Rose Bengal-positive and 43 negative cases. Various diagnostic methods were employed, including specific IgM and IgG antibodies, culture, polymerase chain reaction (PCR), and gamma interferon targeting the brucella antigen. Results: Patients with positive results exhibited symptoms validated by PCR. The gamma interferon assay demonstrated 79.2% sensitivity and 100% specificity. PCR showed 89.3% sensitivity and 100% specificity, while brucella-specific IgG had 21.4% sensitivity and 100% specificity. Other laboratory studies, including Brucella-specific IgM, showed 100% sensitivity and specificity, with positive Brucella culture observed in only one case. Conclusion: Chronic brucellosis's sustained high levels of gamma interferon may offer better follow-up for patients with lingering symptoms than Brucella-specific IgG, especially after acute cases with low-grade fever.

Etiological diagnostic methods and research progress of forest encephalitis

Forest encephalitis is a natural focal disease transmitted through the bite of hard ticks, and its pathogen is the tick-borne encephalitis virus from the Flaviviridae family. The mortality rate of forest encephalitis is relatively high, making laboratory testing significant in diagnosing this disease. This article elaborates on the etiological diagnostic methods and recent research progress in forest encephalitis. Laboratory tests for forest encephalitis mainly include routine examinations, serological tests, virus isolation, and molecular biological testing. The detection of serum-specific IgM antibodies against the forest encephalitis virus is of great importance for early diagnosis, and specific IgG antibodies serve as a "gold standard" for differentiation from other diseases. Techniques such as enzyme-linked immunosorbent assay (ELISA) or indirect immunofluorescence assay for detecting specific IgM antibodies in serum and/or cerebrospinal fluid, the serum hemagglutination inhibition test or serum complement fixation test, and the double serum hemagglutination inhibition test or complement fixation test all contribute to the early diagnosis. The development of molecular testing methods is rapid, and techniques such as metabolomics, digital PCR, and matrix metalloproteinases are also applied in the early diagnosis of forest encephalitis.

Systemic and mucosal immune responses in red tilapia (Oreochromis sp.) following immersion vaccination with a chitosan polymer-based nanovaccine against Aeromonas veronii

A mucoadhesive chitosan polymer-based nanoplatform has been increasingly recognized as an effective mucosal vaccine delivery system for fish. The present study aimed to investigate the effectiveness of immersion vaccination with a chitosan polymer-based nanovaccine to elicit an immune response in serum and mucus of red tilapia and evaluate its protective efficacy after immersion challenge with a heterogenous strain of Aeromonas veronii UDRT09. Six hundred red tilapia (22 ± 1.8 g) were randomly allocated into four experimental groups: control, empty-polymeric nanoparticle (PC), formalin-killed vaccine (FKV), and chitosan polymer-based nanovaccine (CS-NV) in triplicate. The specific IgM antibody levels and their bactericidal activity were assessed in serum and mucus for 28 days after immersion vaccination and followed by immersion challenge with A. veronii. The immersion vaccine was found to be safe for red tilapia, with no mortalities occurring during the vaccination procedure. The specific IgM antibody levels and bactericidal activity against A. veronii in both serum and mucus were significantly higher in red tilapia vaccinated with CS-NV compared to the FKV and control groups at all time points. Furthermore, the serum lysozyme activity, ACH50, and total Ig levels demonstrated a significant elevation in the groups vaccinated with CS-NV compared to the FKV and control groups. Importantly, the Relative Percentage Survival (RPS) value of the CS-NV group (71 %) was significantly higher than that of the FKV (15.12 %) and PC (2.33 %) groups, respectively. This indicates that the chitosan polymer-based nanovaccine platform is an effective delivery system for the immersion vaccination of tilapia.

Immune response and protective efficacy of Streptococcus agalactiae vaccine coated with chitosan oligosaccharide for different immunization strategy in nile tilapia (Oreochromis niloticus)

In the past decade, the outbreak of Streptococcus agalactiae has caused significant economic losses in tilapia farming. Vaccine immunization methods and strategies have gradually evolved from single-mode to multi-mode overall prevention and control strategies. In this study, an inactivated vaccine of S. agalactiae with a chitosan oligosaccharide (COS) adjuvant was constructed using different administration methods: intraperitoneal injection (Ip), immersion combined with intraperitoneal injection (Im + Ip), immersion combined with oral administration (Im + Or), and oral administration (Or). Safety analysis revealed no adverse effects on tilapia, and the vaccine significantly promoted fish growth and development when administered through Im + Or or Or immunization. Following vaccination, innate immunity parameters including SOD, ACP and CAT activities were all significantly enhanced. Additionally, specific serum IgM antibodies reached their highest level at the 6th week post vaccination. Skin and intestinal mucus IgT antibodies reached peaked at the 6th and 7th week post vaccination, respectively. The relative peak expression values for IL-8, IL-12, MHC-I, MHC-II, IgM, IgT, CD4, CD8, TNFα, IFNγ from Im + Ip group were significantly higher than those in Ip group, Im + Or group and Or group in most cases (p < 0.05). Importantly, the relative protection survival of Im + Ip group was the highest (78.6%), followed by the Ip group (71.4%), the Or group (64.3%) and the Im + Or group (57.1%). In summary, this study encourages further research on multi-channel immunization strategies of other kinds of vaccines in other aquatic economic animals to improve their disease resistance.

Investigation of Human Parvovirus B19 Seroprevalence among Pregnant Women Experiencing Spontaneous Abortion in Kirkuk City, Iraq

Human parvovirus B19 (HPV B19) may cross the placenta from mother to the fetus and cause major problems like non-immune hydrops fetalis, severe fetal anemia, as well as fetal mortality. The aim of the study was to determine the seroprevalence of HPVB19 in pregnant women with spontaneous abortion in kirkuk. From November 2022 to the April 2023, blood samples from 135 aborted women aged 15-45 and 45 women with successful pregnancies as a control group were taken. Using the ELISA Technique, the sera from the samples were tested for HPV B19 -specific IgM and IgG antibodies. The result showed that 13 (9.63%) tested positive for HPV B19 IgM, 28 (20.74%) tested positive for IgG,P&lt;0.01.The highest seroprevalence of HPV B19 IgM antibodies was observed in the 15-25 age group (61.54%). HPV B19 IgG antibodies highest prevalence in the 26-35 age groups (50%) (P&lt;0.05).The study showed that in the 1st trimester, 11 cases (84.62%) tested positive for HPV B19 IgM antibodies, while 20 cases (71.43%) tested positive for IgG antibodies.Among women with three abortions, a higher proportion tested positive for HPV B19 antibodies, with 76.92% testing positive for HPV B19IgM and 82.14% testing positive for IgG, among women residing in rural areas, a higher proportion tested positive, with 76.92% testing positive for HPV B19 IgM antibodies and 75% testing positive for HPV B19 IgG antibodies. In the normal weight category, 69.23% tested positive for HPV B19 IgM antibodies, and 82.14% tested positive for IgG antibodies. Aborted women have a higher prevalence of Parvovirus HPV B19 IgM and IgG than control groups, suggesting a potential association between B19 infection and pregnancy loss.