Somatic Antigens Research Articles

Study of allergic reaction to somatic antigen made from a live plague vaccine of the EV strain

Study of allergic reaction to somatic antigen made from a live plague vaccine of the EV strain

Occurrence of carbapenemase producing NAG Vibriocholerae (NAG-CPV.c) in aquatic environment before the recommendation of carbapenem antibiotics

Recent research updates and advances have shown an upsurge of antibiotic resistance reports amongst bacterial species with increasing spread/distribution especially in the water nexus. Such has been the trend among Vibrio cholerae strain as it is observed to be emerging with diverse resistant determinants. The current study presents the occurrence of carbapenemase producing NAG Vibriocholerae (NAG-CPV.c) in aquatic environment before the recommendation of Carbapenem antibiotics. It focused on carbapenem resistant phenotype/genotype among environmental and somatic antigen nonagglutinating V. cholerae (SANAG-Vc) strains recovered from water sources, applying standard microbiological, serological and molecular biology techniques. Domestic water samples were collected for isolation of V. cholerae strains in Eastern Cape Province, South Africa. Carbapenem and specific β-lactamase inhibitors were employed for antibiotic susceptibility testing using K-B disc diffusion technique in addition to the Modified-Hodge-Test (MHT). Our results revealed 61 strains of environmental and SANAG-Vc serogroups. Amongst these confirmed SANAG-Vc strains recovered, 25 possess carbapenemase phenotype or NDM-1 phenotype (40.89%), whereas 24 (39.34%) were MHT positive phenotype. Further gene-based detection revealed 20 (32.79%) PCR confirmed as NDM-1 resistant gene positive strains. It is important to note that the carbapenem members of antibiotics are not readily employed in the therapeutic control of cholera cases as recommended by CLSI. Observing such resistant phenotypes/genotypes indicates a possible transfer/dissemination and emergence of such resistant determinants in the environment as the coastal water serves as a potential breeding hub for such resistant genes among potential pathogens. This is a serious threat to environmental wellness/public health especially those in the sub-urban and poor living localities, who source the analyzed water as their major source of water for domestic activities. The need for adroit and continuous monitoring of released water effluents of domestic and clinical sources remains a control strategy for environmental wellness and water bodies safety.

Evaluation of Somatic Antigens of Adult Toxocara helminthes for Detection of Human Toxocariasis.

We aimed to develop an indirect enzyme-linked immunosorbent assay (ELISA) to evaluate the presence of specific IgG against Toxocara canis and Toxocara cati somatic antigens on the serum of patients with toxocariasis. The sensitivity, specificity, positive and negative predictive values for indirect-ELISA were calculated by receiver operating characteristic curve (ROC) analysis and Youden's J using Likelihood ratio. All statistics were analysed and graphs are plotted using GraphPad Prism version 8.4.3 (Graph Pad Software, La Jolla, CA, USA), with 95% confidence interval (CI). The sensitivity, specificity, positive and negative predictive values for T. canis were 100%, 82%, 79% and 100%, respectively. The mentioned variables for T. cati were 97%, 82%, 78% and 98%, respectively. Five immune reactive bands of 38, 40, 72, 100 and 250 kDa were common in both species. Toxocara crude antigens were highly immunogenic in human sera. Immunoreactive bands against T. canis compared to T. cati somatic antigen were about two times more. Unlike Toxocara excretory-secretory antigen, that was homologue in two species, somatic antigens of T. canis and T. cati showed different immunoreactive bands in our western blot.

Analysis of Protection and Immune Response against Teladorsagia circumcincta in Goats Immunised with Thiol-Binding Proteins from Adult Worms.

In view of the increasing occurrence of anthelmintic-resistant strains of gastrointestinal nematodes in ruminants, various alternative control strategies have been investigated, such as those based on the induction of protective immune responses by immunisation with parasite antigens. In this study, the protective activity of somatic antigens from adult worms of Teladorsagia circumcincta purified by affinity chromatography on thiol-sepharose was analysed in goats. After challenge, the enriched products induced a slight reduction in the cumulative faecal egg counts (21%) and in the number of worms (23.3%), with a greater effect on female worms, which also showed a reduction in parameters related to their fertility. These parasitological findings were associated with a Th2 immune response, with a prominent local humoral response and an eosinophilic infiltrate in the gastric mucosa (negatively associated with the fertility of female worms and the number of worms, respectively), as well as an infiltration of MCHII+, CD4+, IgG+ and IgA+ cells. However, several analyses showed an increase in CD8+ cells in the mucosa, as well as IL-2 expression in the gastric lymph nodes, which may have been associated with inhibition of protective responses or with the development of mixed Th1/Th2 responses, a finding that should be analysed in future studies.

The effect of hydatid cyst protoscolex somatic antigens on full-thickness skin wound healing in mouse.

Wound healing has evolved in recent years, resulting in diverse therapeutic options. This study evaluated the effects of the somatic antigen of the hydatid cyst protoscolex on wound healing in mice with full-thickness skin wounds. Fifty-four adult mice, weighing 25±5g and approximately 60 days old, were divided into three groups (A, B, and C), each further divided into three subgroups. Subgroups A1, A2, and A3 were assigned negative controls. B1, B2, and B3 received hydatid cyst somatic antigen tests at 10µg/SC, whereas C1, C2, and C3 received somatic antigen tests at 20µg/SC. Under general anesthesia, a wound biopsy puncture of 9.8mm in diameter was performed on the mice's back and spine. In the experimental group, antigen and alum adjuvant were administered subcutaneously around the wound, while the control group received Phosphate-Buffered Saline (PBS). Using digital images, a geometric assessment was conducted on days 0, 1, 3, 6, 9, 12, 15, 18, and 21 post-wounding. The obtained images were analyzed by Image J software and after analyzing the data by SPSS software. A significant difference in terms of epithelization was observed in the antigen treatment group with a dose of 20µg on days 3 and 6 (P<0.05). Furthermore, the 20µg antigen group was significantly higher than the 10µg antigen group in terms of this factor on day 3 (P<0.05). Skin samples were taken from all wounds on days 3, 10 and 21 for microscopic evaluation. Regarding epithelization, on day 10, a significant difference was observed in the treatment group with a concentration of 10µg with the control group and the treatment group with a concentration of 20µg (P<0.05). Based on the results of the present study, it can be concluded that somatic antigens of protoscolex hydatid cyst are dose-dependent and antigens with a dose of 20µg by subcutaneous injection accelerate wound healing and epithelialization.

Blackleg: A Review of the Agent and Management of the Disease in Brazil.

The genus Clostridium is an important group of pathogenic and nonpathogenic Gram-positive anaerobic bacteria with a sporulation capacity and wide distribution in different environments, including the gastrointestinal tracts of healthy and diseased animals and humans. Among the pathogenic species of the genus, Clostridium chauvoei stands out as a histotoxic agent. It causes significant myonecrosis such as blackleg, a disease with high lethality, especially in young cattle, and is responsible for significant livestock losses worldwide. The pathogenicity of the disease is complex and has not yet been fully elucidated. Current hypotheses cover processes from the initial absorption to the transport and deposition of the agent in the affected tissues. The virulence factors of C. chauvoei have been divided into somatic and flagellar antigens and soluble antigens/toxins, which are the main antigens used in vaccines against blackleg in Brazil and worldwide. This review provides important information on the first and current approaches to the agent C. chauvoei and its virulence factors as well as a compilation of data on Brazilian studies related to blackleg.

Application of a recombinant novel trypsin from Trichinella spiralis for serodiagnosis of trichinellosis

BackgroundThe excretory/secretory (ES) antigen of Trichinella spiralis muscle larvae (ML) is currently the most widely used diagnostic antigen to detect T. spiralis infection. However, this antigen has certain drawbacks, such as a complicated ES antigen preparation process and lower sensitivity during the early phase of infection. The aim of this study was to investigate the features of a novel T. spiralis trypsin (TsTryp) and evaluate its potential diagnostic value for trichinellosis.MethodsThe TsTryp gene was cloned and recombinant TsTryp (rTsTryp) expressed. Western blotting and an enzyme-linked immunosorbent assay (ELISA) were performed to confirm the antigenicity of rTsTryp. The expression pattern and distribution signature of TsTryp at various life-cycle stages of T. spiralis were analyzed by quantitative PCR, western blotting and the immunofluorescence test. An ELISA with rTsTryp and ML ES antigens was used to detect immunoglobulins G and M (IgG, IgM) in serum samples of infected mice, swine and humans. The seropositive results were further confirmed by western blot with rTsTryp and ML ES antigens.ResultsTsTryp expression was observed in diverse T. spiralis life-cycle phases, with particularly high expression in the early developmental phase (intestinal infectious larvae and adults), with distribution observed mainly at the nematode outer cuticle and stichosome. rTsTryp was identified by T. spiralis-infected mouse sera and anti-rTsTryp sera. Natural TsTryp protease was detected in somatic soluble and ES antigens of the nematode. In mice infected with 200 T. spiralis ML, serum-specific IgG was first detected by rTsTryp-ELISA at 8 days post-infection (dpi), reaching 100% positivity at 12 dpi, and first detected by ES-ELISA at 10 dpi, reaching 100% positivity at 14 dpi. Specific IgG was detected by rTsTryp 2 days earlier than by ES antigens. When specific IgG was determined in serum samples from trichinellosis patients, the sensitivity of rTsTryp-ELISA and ES antigens-ELISA was 98.1% (51/52 samples) and 94.2% (49/52 samples), respectively (P = 0.308), but the specificity of rTsTryp was significantly higher than that of ES antigens (98.7% vs. 95.4%; P = 0.030). Additionally, rTsTryp conferred a lower cross-reaction, with only three serum samples in total testing positive from 11 clonorchiasis, 20 cysticercosis and 24 echinococcosis patients (1 sample from each patient group).ConclusionsTsTryp was shown to be an early and highly expressed antigen at intestinal T. spiralis stages, indicating that rTsTryp represents a valuable diagnostic antigen for the serodiagnosis of early Trichinella infection.Graphical

Immunomodulatory effect of hepatitis B virus (HBV) antigens and MRSA antigens on immune response

Abstract Background: Methicillin-resistant Staphylococcus aureus is one of the most common human invaders and causes different types of illness worldwide. Objectives: This study aimed to illustrate the immune response of purified human peripheral blood mononuclear cells against somatic antigens of methicillin-resistant S. aureus (MRSA) and to study the immunomodulatory effect of hepatitis B virus (HBV) antigens on this immune response. Materials and Methods: A case–control study was applied on 75 peripheral blood mononuclear cells (PBMCs) samples. This work involved five experimental groups, each group involved 15 samples. Group I involves separated PBMCs (1 × 106 cells/mL) alone as a control; group II involves PBMCs stimulated with killed somatic MRSA antigen; group III of PBMCs were stimulated with HBV vaccine only; group IV involves PBMCs pretreated with HBV vaccine for 48 h, then with killed somatic MRSA antigen; and group V involves PBMCs stimulated with mixed of killed somatic MRSA antigen and HBV vaccine. The immune response against MRSA somatic antigens was assessed by measurement of interferon-γ, interleukin-4, and interleukin-5 by enzyme-linked immunosorbent assay technique. Results: The results indicated a significant increase (P &lt; 0.05) in the mean value of IFN-γ in group II (125.9995 pg/mL) and III group (133.6504 pg/mL) and group IV (132.0271 pg/mL) as compared with control (group I) (49.6203 pg/mL). Also, the results showed no significant differences between control group I and group V (49.6203 and 9.6087 pg/mL). IL-4 and IL-5 concentration results revealed a significant increase in all treated groups as compared with control untreated group I. Conclusion: Immune response represented by regulatory cytokines against MRSA is significantly induced by somatic antigens.

Clinical and Microbiological Risk Factors for 30-Day Mortality of Bloodstream Infections Caused by OXA-48-Producing Klebsiella pneumoniae.

Bloodstream infections (BSI) caused by carbapenem-resistant Klebsiella pneumoniae are associated with high morbidity and mortality, and the therapy options available for their treatment are frequently scarce. The aim of this study was to analyze risk factors for 30-day mortality in patients with BSI caused by OXA-48-producing K. pneumoniae. The clinical and treatment features of the patients, who attended a single hospital over a five-year period, were retrospectively reviewed. The microbiological features, including the sequence types (ST) and the somatic (O) and capsular (K) antigens, as well as their resistance properties, comprising phenotypes and genetic background, were also considered. To identify the risk factors for 30-day mortality, uni- and multivariate statistical analyses were performed. The univariate analysis revealed statistically significant correlations for age, male gender, lower respiratory system infection, infection by ST147 isolates, and infection by isolates expressing the K64 antigen. The multivariate analysis, applied to variables yielding p-values close to or lower than 0.05 in the univariate analysis, confirmed gender, lower respiratory system infection, and infection with ST147 isolates, but not age or infection with K64 isolates, as risk factors for 30-day mortality. Moreover, the multivariate analysis showed that patients suffering from hematological malignancies or having been treated with inappropriate therapy, both having p-values slightly higher than 0.05 in the univariate analysis, exhibited significantly poorer outcomes in the multivariant analysis. The association of the ST147 clone with an increased risk of mortality is a novel finding that deserves further attention. Studies like the one presented here can certainly benefit the management of patients with nosocomial BSI caused by carbapenemase-producing K. pneumoniae.

Utilizing immunogenomic approaches to prioritize targetable neoantigens for personalized cancer immunotherapy.

Advancements in sequencing technologies and bioinformatics algorithms have expanded our ability to identify tumor-specific somatic mutation-derived antigens (neoantigens). While recent studies have shown neoantigens to be compelling targets for cancer immunotherapy due to their foreign nature and high immunogenicity, the need for increasingly accurate and cost-effective approaches to rapidly identify neoantigens remains a challenging task, but essential for successful cancer immunotherapy. Currently, gene expression analysis and algorithms for variant calling can be used to generate lists of mutational profiles across patients, but more care is needed to curate these lists and prioritize the candidate neoantigens most capable of inducing an immune response. A growing amount of evidence suggests that only a handful of somatic mutations predicted by mutational profiling approaches act as immunogenic neoantigens. Hence, unbiased screening of all candidate neoantigens predicted by Whole Genome Sequencing/Whole Exome Sequencing may be necessary to more comprehensively access the full spectrum of immunogenic neoepitopes. Once putative cancer neoantigens are identified, one of the largest bottlenecks in translating these neoantigens into actionable targets for cell-based therapies is identifying the cognate T cell receptors (TCRs) capable of recognizing these neoantigens. While many TCR-directed screening and validation assays have utilized bulk samples in the past, there has been a recent surge in the number of single-cell assays that provide a more granular understanding of the factors governing TCR-pMHC interactions. The goal of this review is to provide an overview of existing strategies to identify candidate neoantigens using genomics-based approaches and methods for assessing neoantigen immunogenicity. Additionally, applications, prospects, and limitations of some of the current single-cell technologies will be discussed. Finally, we will briefly summarize some of the recent models that have been used to predict TCR antigen specificity and analyze the TCR receptor repertoire.

Occurrence of a New Variant of Salmonella Infantis Lacking Somatic Antigen.

Salmonella Infantis is one of the most frequent serovars reported in broilers and is also regularly identified in human salmonellosis cases, representing a relevant public health problem. In the laboratories of the Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata (IZSPB), six Salmonella Infantis strains with antigenic formula -:r:1,5 have been isolated from the litter and carcass of broilers between 2018 and 2022. The strains were investigated to evaluate their phenotype, antibiotic resistance and genomic profiles. Genomic analysis confirmed that the isolates belonged to the Infantis serotype and to the sequence type ST32. Moreover, all strains showed a multidrug-resistant (MDR) profile and were characterised by the presence of the IncFIB plasmid incompatibility group. Three strains had the blaCTX-M-1 gene, and one of them carried IncX1. The presence of this new variant of S. Infantis is particularly relevant because it could expand the landscape of the S. Infantis population. The absence of the somatic antigen could pose a problem in both isolation and serotyping and a consequent public health concern due to the spread of Salmonella infection.

Secretory IgA as Biomarker for Gastrointestinal Nematodes Natural Infection in Different Breed Sheep.

Specific IgA antibody has been shown to play an important role in resistance to gastrointestinal nematode (GIN) infections in sheep, particularly in Teladorsagia circumcincta parasitosis. In some breeds, negative associations have been shown between IgA levels and worm burden in experimentally infected sheep. In the present study, we have studied the relationship between IgA levels in naturally infected sheep (582 ewes in total; 193 younger than one year old and 389 older than one year old) and fecal egg count (FEC) in the Assaf, Castellana, and Churra breeds. ELISA assays were performed to measure IgA levels against the somatic antigen of T. circumcincta third larval stage (L3) and a 203-amino-acid fragment of the protein disulfide isomerase from the same GIN species. A multilevel random intercept model was developed to predict the infection risk according to age or breed. Spearman's correlation rank was used for statistical analysis. The prediction model showed that breed was not an influential factor in this study, although the Assaf breed could be considered slightly more susceptible than the others. In addition, age affected the infection risk, with the young ewes more susceptible to infection than the adult groups, except for the Castellana breed, whose risk of infection was similar at all ages. The most significant positive association was found between FEC and IgA measured in the nasal secretions of young ewes using both antigens (Rho = 0.5; p = 0.00); the correlation of FEC with IgA in serum was moderately significant (Rho = 0.306; p = 0.00). Comparing both antigens, the protein disulfide isomerase antigen was less reactive than the somatic antigen from L3. In conclusion, under natural conditions, specific IgA against GIN was positively associated with FEC in sheep, with nasal secretions from young animals being the sample where this association is stronger, which, therefore, could be used as a marker of infection in further studies.

The mouse model of chronic asthma: Airway remodelling and disease exacerbation by somatic antigen of Echinococcus granulosus.

There is now sufficient evidence to support an inverse association between helminth infection and secreted products with allergic/autoimmune disorders. Accordingly, several experimental studies have shown that Echinococcus granulosus infection and hydatid cyst compounds are able to suppress immune responses in allergic airway inflammation. This is the first study on effects of somatic antigens of E. granulosus on chronic allergic airway inflammation in BALB/c mice. Mice in OVA group were intraperitoneally (IP) sensitized with OVA/Alum. Subsequently, were challenged by nebulizing of OVA 1%. The treatment groups received somatic antigens of protoscoleces on the specified days. Mice in PBS group were received PBS in both sensitization and challenge. The effects of somatic products on development of chronic allergic airway inflammation were evaluated by examining histopathological changes, the recruitment of inflammatory cells in the bronchoalveolar lavage, cytokines production in the homogenized lung tissue, and total antioxidant capacity in serum. Our findings show that the co-administration of somatic antigens of protoscoleces simultaneously with the development of asthma intensifies allergic airway inflammation. The identification of effective components involved in exacerbation of allergic airway inflammation manifestations will be a crucial approach to understanding the mechanism of these interactions.

Anti-Tumor Effect of Protoscolex Hydatid Cyst Somatic Antigen on Inhibition Cell Growth of K562.

Today, cancer is one of the most important causes of death in the world, and so far, many treatment methods have been used in this field. Immunotherapy is considered one of the newest developments in this science, and it is still being investigated in some forms in different cancers and with a variety of antigens as well. One of the subsets of cancer immunotherapy is its treatment using parasitic antigens. The present study evaluated the effect of using somatic antigens of protoscoleces of Echinococcus granulosus on K562 cancer cells. In this study, hydatid cysts' protoscolex antigens were extracted, purified, and added to K562 cancer cells at three concentrations (0.1, 1, and 2mg/ml) and on three times (24, 48, and 72h). The number of apoptotic cells was compared to the control flask. The antigen concentration of 2mg/ml was used as a control sample to investigate its cytotoxic effect on the growth of healthy HFF3 cells. Annexin V and PI tests were also performed to differentiate apoptosis from necrosis. In flasks treated with hydatid cyst protoscolex antigen, all three concentrations significantly reduced the growth of cancer cells compared with the control flask, and concentration 2 of crude antigen significantly caused the death of cancer cells. Furthermore, more cancer cells underwent apoptosis by increasing the time of exposure to the antigen. On the other hand, flow cytometry results also showed that the amount of apoptosis has increased compared to the control group. In fact, Protoscolex hydatid cyst somatic antigens induce programmed cell death in K562 cancer cells while not having a cytotoxic effect on normal cells. Therefore, it is suggested to do more research on the anti-cancer and therapeutic properties of the antigens of this parasite.

Effect of somatic antigens of Dirofilaria repens adult worms on angiogenesis, cell proliferation and migration and pseudo-capillary formation in human endothelial cells

BackgroundAngiogenesis is defined as the formation of new vessels by sprouting of endothelial cells from pre-existing vessels in response to stimuli, such as hypoxia or inflammation. Subcutaneous dirofilariasis, caused by Dirofilaria repens, is a zoonotic disease characterized by the formation of subcutaneous nodules with the presence of at least one encapsulated worm, showing perivascular vascularization around it. The aim of this study is to analyze whether the somatic antigen of adult D. repens worms interacts with and modulates the angiogenic mechanism, cell proliferation and migration, and formation of pseudo-capillaries.MethodsThe expression of VEGF-A, VEGFR-1/sFlt, VEGFR-2, mEnd and sEnd in cultures of human vascular endothelial cells stimulated with somatic antigen of adult worms of D. repens (DrSA), vascular endothelial growth factor (VEGF) and DrSA + VEGF were evaluated by using ELISA commercial kits. Cellular viability was analyzed by live cell count, cytotoxicity assays by using a commercial kit, cell proliferation by MTT-based assay, cell migration by wound-healing assay carried out by scratching wounds and capacity of formation of pseudo-capillaries analyzing cell connections and cell groups in Matrigel cell cultures. In all cases unstimulated cultures were used as controls.ResultsDrSA + VEGF significantly increased the expression of VEGF-A, VEGFR-2 and mEndoglin compared to other groups and unstimulated cultures. Moreover, DrSA + VEGF produced cell proliferation and migration and increased the formation of pseudo-capillaries.ConclusionsSomatic antigen of adult D. repens worms activated the proangiogenic mechanism, cell proliferation and cell migration as well as formation of pseudo-capillaries in this in vitro human endothelial cell model. These processes could be related to the survival of adult D. repens in subcutaneous nodules in infected hosts.Graphical

Clonal evolution and stereotyped sequences of human IgE lineages in aeroallergen-specific immunotherapy

Allergic disease reflects specific inflammatory processes initiated by interaction between allergen and allergen-specific IgE. Specific immunotherapy (SIT) is an effective long-term treatment option, but the mechanisms by which SIT provides desensitization are not well understood. Our aim was to characterize IgE sequences expressed by allergen-specific B cells over a 3-year longitudinal study of patients with aeroallergies who were undergoing SIT. Allergen-specific IgE-expressing clones were identified by using combinatorial single-chain variable fragment libraries and tracked in PBMCs and nasal biopsy samples over a 3-year period with antibody gene repertoire sequencing. The characteristics of private IgE-expressing clones were compared with those of stereotyped or "public" IgE responses to the grass pollen allergen Phleum pratense (Phl p) 2. Members of the same allergen-specific IgE lineages were observed in nasal biopsy samples and blood, and lineages detected at baseline persisted in blood and nasal biopsy samples after 3 years of SIT, including B cells that express IgE. Evidence of progressive class switch recombination to IgG subclasses was observed after 3 years of SIT. Acommon stereotyped Phl p 2-specific antibody heavy chain sequence was detected in multiple donors. The amino acid residues enriched in IgE-stereotyped sequences from seropositive donors were analyzed with machine learning and k-mer motif discovery. Stereotyped IgE sequences had lower overall rates of somatic hypermutation and antigen selection than did single-chain variable fragment-derived allergen-specific sequences or IgE sequences of unknown specificity. Longitudinal tracking of rare circulating and tissue-resident allergen-specific IgE+ clones demonstrates persistence of allergen-specific IgE+ clones, progressive class switch recombination to IgG subtypes, and distinct maturation of a stereotyped Phl p 2 clonotype.

Secretory Excretory and Somatic Immunogenic Antigens Profiles of Adult Fasciola spp.

Fascioliasis is a common human-animal disease that is reported in most parts of the world. Fascioliasis is also prevalent in different provinces of Iran. Since it has done no study on the excretory/secretory and somatic immunogenic antigens profiles of adult Fasciola in Iran, the present study was performed on the Fasciola spp. collected from Mazandaran province. For this purpose, the Fasciola worm was isolated from the liver of infected sheep, then its excretory/secretory and somatic antigens were prepared from adult worms. The protein of the samples was measured by the Lowry method. Then, somatic and secretory excretions were examined by SDS-PAGE and the protein profile of the two substances was determined. To evaluate the immunogenicity, the somatic and secretory excretions antigens of Fasciola spp. were injected into white rabbits and after boosting, the blood serum of the rabbits was collected and then Western blotting was performed on them and the results were evaluated. According to the results of Western blotting, 11 somatic antigen bands with a molecular weight of 149, 122, 99, 85, 75, 65, 50, 46, 40, 37, 30 kDa and 12 protein bands of excretory/secretory antigens with molecular weights of 100, 82, 75, 70, 58, 55, 47, 40, 38, 37, 30,25 kDa were observed in adult Fasciola spp. that immunogenic, which appear to have a protective effect or can be used to prepare a diagnostic kit.

Seroprevalence of nasal myiasis in camels determined by indirect enzyme-linked immunosorbent assay utilizing the most diagnostic Cephalopina titillator larval antigens.

Nasal myiasis is a serious parasitic disease among camels caused by Cephalopina titillator larvae that negatively affect animal health and production globally. The diagnosis of the infestation relies on postmortem examination of the head region, which considers a cause impeding treatment of live animals and may be misdiagnosed as central nervous system disorders. This study aimed to identify the most diagnostic larval antigen with the capacity for monitoring C. titillator infestation, and to estimate the seroprevalence of nasal myiasis in camels in Egypt, using indirect enzyme-linked immunosorbent assay (ELISA). Three hundred and six male camels of Egyptian and Sudanese breeds, aged 2-5 years, were clinically evaluated for respiratory and/or nervous disorders in Cairo Governorate, Egypt. At the time of slaughter, blood samples were collected from all examined animals. The postmortem examination of 38 animals was conducted. Salivary glands, hemolymph, and somatic antigens were extracted from the second and third larval instars. The results revealed that the salivary gland antigen was the most potent antigen in detecting C. titillator specific total IgG antibodies compared to haemolymph and crude somatic antigens. Using receiver-operating characteristic curves and area under the curve, the salivary gland antigen had a sensitivity of 91.67% and a specificity of 92.31%, respectively. It has the highest positive predictive value, 95.7%, and negative predictive value, 85.7%. However, using somatic and hemolymph antigens revealed a sensitivity of 79.17% and 70.83% and a specificity of 76.9% and 84.6%, respectively. There was complete concordance between ELISA results and autopsy findings (true positive). One hundred and forty out of 306 (45.8%) camel serum samples were found to contain C. titillator. This study demonstrated that salivary gland antigen is more effective than somatic and hemolymph antigens in accurately detecting nasal myiasis in camels. In addition, determining the seroprevalence of nasal myiasis with the salivary gland antigen through indirect ELISA revealed that it is a prevalent disease among camels in Egypt. Periodic surveillance of the C. titillator prevalence is necessary for effective management and control measures.

Seroprevalence of Fasciola-infection using Enzyme-linked immunosorbent assay in large ruminants from Pakistan

Fasciolosis, caused by liver fluke species of the genus Fasciola, are well recognized because of its high veterinary impact. Stool examination for Fasciola eggs is not a sensitive method, and limited efforts to find a reliable and cheaper means of detection are available. The present study aimed to develop rapid diagnostic ELISA test against fasciolosis. The excretory/secretory (ES) and somatic (SA) products of Fasciola helminths were analysed using polyacrylamide gel electrophoresis (PAGE). Immunogenicity was evaluated by immunoblotting using hyperimmune sera raised in rabbits and seroprevalence was determined by indirect ELISA. The results of SA antigen of Fasciola species showed polypeptide bands ranged from 10kDa-100kDa, while ES antigen of Fasciola showed bands of 15kDa-55kDa. The immunoblotting results showed the most prominent bands against ES antibodies were 25, 35, 55-70, 100 and 250 kDa and SA antigens showed 10, 15-25, 35, 70, 100 and 250 kDa polypeptide bands. The sensitivity and specificity of developed indirect ELISA for SA antigens was 95.45% and 87.1%, while for ES antigens was 100% and 77.42% respectively. The overall seroprevalence recorded for fascioliasis based on SA antigen was 39.8% and 29.8% for ES antigen. The fasciolosis did not show significant association with host type, sex and age groups of examined animals, however significantly higher infection was found in months of September and October. The result provides sensitive in house immunodetection assay for diagnosis of fasciolosis alternative to commercial kits with high import cost.

Effects of adult Ascaris suum and their antigens (total and trans-cuticular excretory-secretory antigen, cuticular somatic antigen) on intestinal nutrient transport in vivo.

AbstractAscaris suumconstitutes a major problem in commercial pig farming worldwide. Lower weight gains in infected pigs probably result from impaired nutrient absorption. This study investigated intestinal nutrient transport in 4 groups of 6 pigs each, which were inoculated with 30 living adultA. suum, or antigen fractions consisting of (1) total excretory–secretory (ES) antigens of adult worms, (2) ES antigens secreted exclusively from the parasites' body surface (trans-cuticular ES) and (3) cuticular somatic antigens ofA. suum, compared to placebo-treated controls. Three days after inoculation into the gastrointestinal tract, glucose, alanine and glycyl-l-glutamine transport was measured in the duodenum, jejunum and ileum using Ussing chambers. Transcription of relevant genes [sodium glucose cotransporter 1 (SGLT1), glucose transporter 1 (GLUT1), GLUT2, hypoxia-inducible factor 1-alpha (Hif1α), interleukin-4 (IL-4), IL-13, signal transducer and activator of transcription 6 (STAT6), peptide transporter 1 (PepT1)] and expression of transport proteins [SGLT1, phosphorylated SGLT1, GLUT2, Na+/K+-ATPase, amino acid transporter A (ASCT1), PepT1] were studied. Although no significant functional changes were noted after exposure to adultA. suum, a significant downregulation of jejunal GLUT1, STAT6, Hif1αand PepT1 transcription as well as ileal GLUT2 and PepT1 expression indicates a negative impact of infection on transport physiology. Therefore, the exposure period of 3 days may have been insufficient for functional alterations to become apparent. In contrast,A. suumantigens mainly induced an upregulation of transport processes and an increase in transcription of relevant genes in the duodenum and jejunum, possibly as a compensatory reaction after a transient downregulation. In the ileum, a consistent pattern of downregulation was observed in all inoculated groups, in line with the hypothesis of impaired nutrient transport.