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Monoclonal antibodies (mAbs) against bovine lymphocyte cell surface antigens namely, MHC Class I, MHC class II (DP, DQ and DR), CD3, CD4, CD8, gamma delta TCR, WC1N1 and WC1N2, were tested for their reactivity on apparently normal buffalo mononuclear cells prepared from spleen, lymph nodes and peripheral blood. All the mAbs cross-reacted with the buffalo mononuclear cells. The mean (±SD) CD4:CD8 cell ratio in the peripheral blood of apparently normal buffaloes was 1.08 ± 0.049 while in the spleen and lymph nodes it was 0.90 ± 0.080 and 1.81 ± 0.430, respectively. The lymphocyte subsets in the buffaloes positive for tuberculosis by the single intra dermal (SID) test was found to be altered; the CD4 cells were reduced while the CD8 and gamma delta cells were increased. The mean CD4:CD8 ratio in the SID positive buffaloes was 0.36 ± 0.010. [ABSTRACT FROM AUTHOR]

Abstract: The neutralising antibody response after the yellow fever vaccine (YF-17D) skin test was measured in 7 egg allergic persons in whom further vaccination was abandoned because of a strong local urticarial reaction to the YF-17D vaccine test dose. We found that this test dose of 0.1mL of YF-17D vaccine was sufficient to induce a protective antibody response in all 7 subjects. Intradermal injection of 1/5th dose of the yellow fever vaccine appears to be sufficient, in non-allergic as well as allergic persons, and non-inferior to the subcutaneous full dose. [Copyright &y& Elsevier]

• Lung fluke infection is a rare etiology for exudative pleural effusion. • Eosinophilic pleural effusion or eosinophilic infiltration are characteristics. • Certain diet histories are important clues for lung fluke infection. • Positive intradermal tests for Paragonimus -specific antigens are reliable. Objectives: Pleural effusion caused by lung fluke is a rare etiology of exudative pleural effusion (EPE), which is often misdiagnosed or delayed. We aim to summarize the diagnosis and treatment course of EPE caused by lung fluke infection and put forward a practical diagnosis approach. Methods: We retrospectively analyzed the diagnosis and treatment of 14 cases of EPE caused by lung fluke infection diagnosed by enzyme-linked immunosorbent assay of serum antibodies or egg detection. Results: All patients (100%) with an absolute count of eosinophils in peripheral blood exceeded 0.5 × 109/l, and 10 patients (71.4%) had a history of special ingestion. Eosinophilic PE occurred in 11 patients (78.6%), pleural biopsy of medical thoracoscopic demonstrated eosinophils infiltration in nine patients (64.3%), and parasite eggs in one patient. All patients showed positive intradermal tests for Paragonimus -specific antigens and enzyme-linked immunosorbent assay of serum antibodies to Paragonimus. Conclusion: For patients with unexplained PE, lung fluke infection should be highly suspected when pleural fluid or pleural biopsy shows eosinophilic PE or eosinophils infiltration, especially for patients with certain diet history. [ABSTRACT FROM AUTHOR]

Abstract: Immediate-reading skin tests (pricks and intradermal tests) are indicated in patients reporting symtoms suggesting immediate-type hypersensitivity reactions. The diagnostic and/or predictive value of these tests is good with betalactams, latex and myorelaxants, and several other substances such as corticosteroids, dyes, antiseptics, vaccines, etc. Non irritant concentrations have been determined for several substances (betalactams, myorelaxants, etc.), but are unknown for numerous other substances. For other substances, non irritant concentrations in skin tests should be evaluated in control patients, in parralel with skin tests performed in patients. The diagnostic value of nonimmediate-reading skin tests is highly variable, and depends on the drug and the nature of the symptoms. Although the sensitivity and the specificity of nonimmediate-reading skin tests are not perfect, these tests are useful in the diagnosis of mild to moderately severe toxidermias, such as nonimmediate urticarias and angio-oedemas, fixed drug eruptions, maculopapular rashes, and acute generalized exanthematic pustulosis. In contrast, their diagnostic value is low in potentially severe toxidermias. The diagnostic value of responses in intradermal tests is usually higher than in patch-tests. However, patch-tests may be positive in few patients with negative intradermal tests. Ideally, skin tests should be performed between 6 weeks and 1 to 2 years after the clinical reaction. However, most nonimmediate drug sensitizations can be diagnosed several years later. [Copyright &y& Elsevier]

The insulin analogues, aspart and lispro, have been considered safe alternatives for patients with insulin allergy, because of their decreased immunogenicity. However, recent several reports showed that neither of them was completely free from allergic reactions. We also experienced a patient with insulin allergy not only to human regular insulin but also to both of the insulin analogues. Interestingly, the insulin analogues, which readily dissociate from polymer to monomer, induced the most severe allergic reaction among several types of human insulin reagents in the present case. Allergy to crystalline zinc-insulin, the three-dimensional structure of which results in delayed dissociation and absorption, was negative on intradermal tests. However, its large subcutaneous injection caused local allergic reaction. These results suggested that the allergic reaction might depend on the rapidity of insulin monomerization and absorption, and thus that the immunogenic residue of insulin is concealed when insulin is polymerized. Based on the intradermal tests, we speculated that the antigenic epitope might be B30-Thr in the present case. We also report here the modified method of insulin desensitization using crystalline zinc-insulin with prednisolone tapering. This might be a simple and useful treatment for insulin allergy. [Copyright &y& Elsevier]

Background: Roughly 10% of the U.S. population has a documented penicillin (PCN) allergy. Among these individuals, over 95% are able to tolerate PCNs. The inability to use PCNs can result in suboptimal outcomes. In August 2019, Lawrence Memorial Hospital Health System implemented a pharmacist-led PCN allergy testing service to assist with delabeling PCN allergies and increase access to this class of antibiotics. Objectives: The primary objective was to describe the number of patients who underwent PCN allergy testing and were delabeled from PCN allergy. A secondary objective was to report the number of patients who received and tolerated PCN antibiotics after being delabeled from PCN allergy. Methods: This retrospective chart review was conducted during the initial 17 months of a pharmacist-ledPCN allergy testing service. Eligible patients with a history of an immunoglobulin E (IgE)-mediated reaction underwent a 3-step test that consisted of a scratch test, an intradermal test, and an oral challenge. Eligible patients who did not have a history of IgE-mediated reaction underwent a 2-step graded oral challenge. Descriptive statistics were used for data analysis. Results: Between August 2019 and January 2022, 70 patients underwent testing, and 66 patients were delabeled from PCN allergy. Four patients who underwent the 3-step test developed reactions of mild to moderate severity. All patients who underwent the graded oral challenge were delabeled from PCN allergy. The rate of PCN allergy was 5.7%, whereas the rate of type I IgE-mediated reaction was 1.4%. All 23 patients who received an antibiotic from the PCN class after a negative allergy test tolerated the PCN antibiotic without an incident. Conclusion: PCN allergy testing is an effective way to delabel PCN allergies from most patients presenting with a PCN allergy history. Skin testing followed by an oral challenge or a graded oral challenge alone are safe methods for conducting PCN allergy testing in the primary care setting. [ABSTRACT FROM AUTHOR]

Abstract New techniques in equine veterinary allergy testing include multiple allergen simultaneous tests (MASTs) that cover insect bite hypersensitivity (IBH), respiratory and atopic allergic diseases. The in vivo intradermal test (IDT) method is one of the most widely used techniques in veterinary practice, but recently, in vitro methods have been introduced as possible alternatives. This study was designed to assess the correlation between IDTs and MASTs based on IgE determination in serum. The study was performed on Malopolski breed horses with IBH (n = 26). IDTs were performed, and allergen-specific IgE serum concentrations were measured using a monoclonal anti-IgE antibody during the years 2008–2016. An allergen panel containing Tabanus spp., Culicoides nubeculosus , mosquito (Culex spp.), Simulium equinum , and Stomoxys calcitrans was used. This study shows good agreement and correlation (correlation coefficient rho, 0.413–0.589) between the two tests for four (Tabanus spp, Culicoides nubeculosus , Culex spp., Simulium equinum) of the five tested allergens in horses with IBH. The overall sensitivity (Se) of the allergen-specific IgE insect horse panel was 72.6% (range, 30.8%–91.3%), the specificity (Sp) was 63.6% (range, 50%–70.1%), and the accuracy (Acc) was 73.3% (range, 48%–88.5%) compared with IDTs. This study found good agreement and correlation between IDTs and serological MASTs based on monoclonal antibodies for most of the determined allergens for horses with IBH. Highlights • New serological techniques in equine veterinary allergy testing and in vivo intradermal test in horses with insect bite hypersensitivity were compared. • Immunoenzymatic multiple allergen simultaneous test shows good correlation with intradermal test for Tabanus spp, Culicoides nubeculosus , Culex spp., and Simulium equinum. • Multiple allergen simultaneous test has a moderate accuracy, high sensitivity, and moderate specificity for diagnosing insect sensitization in horses. • Multiple simultaneous allergen test can be used in the causative diagnosis of insect allergy to select the relevant allergens for specific immunotherapy. [ABSTRACT FROM AUTHOR]

Suspected allergic reactions after mRNA COVID-19 vaccination withheld multiple individuals from getting fully vaccinated during the pandemic. We vaccinated adults who had experienced possible allergic symptoms after their first intramuscular dose of a COVID-19 mRNA vaccine with a 1/5th fractional intradermal test dose of the mRNA-1273 (Moderna) COVID-19 vaccine. No anaphylactic reactions were observed after intradermal vaccination (n = 56). Serum anti-S1 IgG concentrations were measured using a bead-based multiplex assay four weeks after vaccinations. Antibody concentrations were compared with a previously collected nationwide cohort that had received two intramuscular doses of mRNA-1273. Antibody responses in all subjects tested (n = 47) were comparable to standard of care intramuscular dosing. Fractional intradermal dosing of mRNA COVID-19 vaccines may provide a pragmatic solution that is safe, time efficient compared to skin prick testing, dose sparing and immunogenic in individuals with suspected vaccine allergy. [ABSTRACT FROM AUTHOR]

• Yellow fever vaccine. • Egg hypersensitivity. • Anaphylaxis. • Graded dosing. • Immunologic. Yellow fever vaccine (YFV) is recommended in endemic areas but represents a risk for egg-allergic patients, as it is cultivated in embryonated eggs. This study aims to describe the outcomes of yellow fever vaccination in patients with confirmed egg allergy (EA). Methods:A prospective study was conducted from January 2018 to September 2019. EA was diagnosed through positive oral food challenge (OFC), recent history of anaphylaxis following egg contact (anaphylaxis in the last 6 months) or immediate allergic reaction in the last 2 months with positive specific IgE. A skinprick test (SPT) with YFV was performed. If the SPT was negative, an intradermal test (ID) was performed at a 1:100 dilution. If the ID was negative, a full dose of YFV was administered. If the skin prick test or ID were positive, the YFV was administered using a graded dosing protocol. Results: It was included 58 patients with confirmed egg allergy (36 M:22F), with a median age of 2.3 years (0.7–13.9 y/o). Forty-two patients had a positive OFC. Nine reported recent anaphylaxis. The other 7 had reactions in the last 2 months with positive specific IgE. During OFC, 15 presented anaphylaxis, while the other 27 presented hives and/or angioedema or vomiting. SPT with YFV was negative in all patients. ID was negative in 48 patients who uneventfully received a full dose of YFV. Ten patients had a positive ID and received YFV in graded doses. Six patients presented a mild reaction controlled with antihistamines, and 4 patients received the vaccine without reactions. Positive ID was significantly related to the vaccine reaction (p < 0.0001). Administration of YFV using a specific protocol was safe even in anaphylactic patients. However, we recommend performing the ID, which can help predict a higher risk of vaccine reaction. An appropriate setting is required to control adverse events. [ABSTRACT FROM AUTHOR]

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• Solving the problem of classifying images of tuberculin test results. • CSN-II applied to image segmentation. • Feature enhancement using information entropy and Bhattacharya distance. • Effective prevention of threshold drift in the Otsu method. The tuberculin skin test (TST) is an intradermal test used to diagnose Type IV hypersensitivity reactions caused by Mycobacterium tuberculosis infection. Segmentation of TST result images provides a foundation for large-scale tuberculosis screening and auxiliary diagnosis. This paper presents a specialized method for identifying TST results. Initially, a clustering approach is employed to reduce pixel complexity, followed by a linear transformation using CSN-II to enhance the original RGB space with robust color space properties. Subsequently, high-probability pixel points are located, and their Gaussian kernel convolution range is determined using the Bhattacharyya Distance. Through convolution and iterative feature amplification, the target characteristics are progressively enhanced. Finally, an improved OTSU method is proposed for segmenting TST result images. In this method, an adaptive entropy threshold is utilized to reduce the search range of the OTSU method, enhancing the relative contrast between the target and the background. Moreover, a weighted adjustment is applied to the obtained OTSU threshold to prevent drift towards backgrounds with larger intra-class variances. Experimental results demonstrate that the proposed method achieves higher segmentation accuracy and robustness in TST result image segmentation compared to traditional OTSU methods and other improved approaches, such as the neighborhood valley-emphasis method, logarithmic OTSU, and weighted OTSU, Finally, we calculate a relative value is calculated by dividing the remaining number of segmented pixels by the total number of pixels, we then classify the results based on the relative value and in reference to medical diagnostic standards our method is intended to establish an algorithmic basis for rapid screening and classification of tuberculosis on a large scale. [ABSTRACT FROM AUTHOR]

Summary: Although bovine tuberculosis is widespread throughout Africa, very little is known about risk factors for Mycobacterium bovis infection in either human or cattle populations. A human case–control study was conducted in northern Tanzania, comparing risk factors and prevalence of cattle interdermal test positives of cases (cervical adenitis cases from which M. bovis was isolated) with age- and sex-matched controls (selected at random from potential hospital attendees within the community). A cattle cross-sectional study was also set-up involving 27 villages selected at random in four districts, with 10,549 cattle and 622 herds tested, and questionnaire surveys conducted in 239 households. M. bovis was confirmed in seven of 65 (10.8%) human cervical adenitis cases, of which only one came from a household owning infected cattle. M. bovis in human patients was associated with families in which a confirmed diagnosis of tuberculosis had previously been made () and with households far (>100m) from neighbours (). In cattle, overall prevalence of intradermal test positives was low at 0.9% (0.70–1.06%), but widespread, with 11.8% (8.44–13.17%) herds containing at least one reactor. Prevalence of intradermal test positives increased significantly with cattle age (). Herds with the following risk factors had a significantly greater prevalence of intradermal test positives: >50 cattle in the herd (); herds housed inside at night () and herds in contact with wildlife (). Furthermore, villages that experienced annual flooding had a higher prevalence of infection (). [Copyright &y& Elsevier]

Atopic dermatitis in dogs is a common allergic skin disease that affects substantial numbers of dogs in the UK. The purpose of this study was to compare the results of an intradermal test (IDT) and an in vitro test in a large cohort of dogs. Dogs were intradermal tested with Greer allergens (Greer Labs Inc, Lenoir, NC, USA) using standard techniques. At the same time blood samples were drawn and submitted for evaluation by ELISA using the ALLERCEPT™ Definitive Allergen Panels for allergen-specific IgE, a commercial assay that uses a biotinylated recombinant extracellular domain of the high affinity Fc-epsilon receptor alpha chain protein (Fc&epsiv;RIα). The allergens used in the two tests included grass, tree and weed pollens, moulds, flea saliva/whole flea extract and house dust mite species. The optical density readings from the ELISA for each allergen were compared with the results of the IDT for 265 dogs. The prevalence of positive reactions in the ELISA was equal to or greater than the results of the IDT in the case of almost all of the allergens, but two notable exceptions were the house dust mites Dermatophagoides farinae and Dermatophagoides pteronyssinus. These two allergens were the most common positive reactions by IDT (prevalence D. farinae 78.9%, D. pteronyssinus 66.4%). The results of the two tests were significantly different (McNemar’s test, P<0.05) for 16 of the 22 allergens. The sensitivities of the ELISA compared to the IDT (where there were more than 3 dogs with positive reactions in both tests) varied between 19.3 and 77.1% (D. pteronyssinus 19.3% and D. farinae 67.9%) and the specificities varied between 64.2 and 96.6% (D. pteronyssinus 96.6% and D. farinae 89.3%). [Copyright &y& Elsevier]

The diagnosis of tuberculosis (TB) in goats is based mainly on the single and comparative intradermal tuberculin (SIT and CIT) tests and, exceptionally, on the interferon-gamma (IFN-γ) assay, however they are not perfect in terms of sensitivity and specificity. Nevertheless, various serological assays that provide a potential cost-effective approach for the control of TB are also available or under development, and a variety of results have been reported regarding the ability of these tests to detect infected animals, particularly in the early stages of infection. In the present study, SIT/CIT and IFN-γ tests and three different serological assays were evaluated during two consecutive herd testing events in a recently infected caprine herd (n = 447) with a high prevalence of infection in order to evaluate their performance and provide field data with which to improve the TB control programs in this species. The proportion of infected animals that tested positive among all the infected goats (T+/I+ value) in the last herd testing event ranged from 26.2% (IC95%; 19.3–34.5) to 85.7% (IC95%; 78.5–90.7) using cell-based diagnostic tests. The SIT/SCIT tests detected more infected goats than the IFN-γ test, regardless of the interpretation criteria. The T+/I+ value of serology was 83.2 (IC95%; 75.2–89), although it increased significantly (P < 0.05) when using samples collected 15 days after the intradermal test (100%, IC95%; 97–100). In general, a parallel interpretation of intradermal tests with serology maximized the detection of infected goats. These results demonstrate that serological tests are valuable diagnostic tools to maximize the detection of TB infected goats, even in recent outbreaks, accelerating the eradication process. [ABSTRACT FROM AUTHOR]

Snake bites are a concerning health problem in Egypt and other tropical countries that are effectively managed with anti-snake venom (ASV). ASV has common reactive complications that are usually mild. Rarely, anaphylaxis and severe systemic reactions may occur following ASV. In this case report, we present a rare condition of Kounis syndrome, acute allergic myocardial infarction, in a young man during anaphylaxis following an allergic intradermal test of VACSERA snake antivenom. [Display omitted] • Kounis Syndrome is a rare acute coronary syndrome. • Kounis Syndrome occurs as a consequence of allergic reactions. • Kounis Syndrome is an uncommon and underdiagnosed disease. • Kounis Syndrome may occur as a rare complication of antivenom administration. • Management of Kounis syndrome is challenging and needs careful use of drugs. [ABSTRACT FROM AUTHOR]

The development of new vaccines against animal tuberculosis (TB) is a priority for improving the control and eradication of this disease, particularly in those species not subjected to compulsory eradication programmes. In this study, the protection conferred by the Mycobacterium tuberculo sis SO 2 experimental vaccine was evaluated using a natural infection model in goats. Twenty-six goats were distributed in three groups: (1) 10 goats served as a control group; (2) six goats were subcutaneously vaccinated with BCG; and (3) 10 goats were subcutaneously vaccinated with SO 2 . Four months after vaccination, all groups were merged with goats infected with Mycobacterium bovis or Mycobacterium caprae , and tested over a 40 week period using a tuberculin intradermal test and an interferon-γ assay for mycobacterial reactivity. The severity of lesions was determined at post-mortem examination and the bacterial load in tissues were evaluated by culture. The two vaccinated groups had significantly lower lesion and bacterial culture scores than the control group ( P < 0.05); at the end of the study, the SO 2 vaccinated goats had the lowest lesion and culture scores. These results suggest that the SO 2 vaccine provides some protection against TB infection acquired from natural exposure. [ABSTRACT FROM AUTHOR]

Le paracétamol est un antalgique très utilisé dans la population générale avec peu d'effets indésirables. Cependant, quelques cas d'anaphylaxie à cette molécule ont été rapportés et les explorations allergologiques sont en général négatives. Un patient de 19 ans, atopique, a présenté une urticaire généralisée avec angiœdème du visage, dyspnée puis malaise avec perte de connaissance en novembre 2017, dans l'heure suivant la prise d'un comprimé de paracétamol 1000 mg. Le tableau avait régressé en 30 min après prise d'antihistaminique et prednisolone. Auparavant et depuis cette réaction, le patient tolérait bien les anti-inflammatoires non stéroïdiens. En octobre 2018, le prick-test paracétamol (10 mg/mL) était négatif, ainsi que l'intradermo-réaction (IDR) 10−2 (0,1 mg/mL). Une dose de 50 mg de paracétamol per os a été donnée avec une bonne tolérance. La deuxième série de tests s'est déroulée un mois plus tard : le prick-test et l'IDR 10−2 étaient positifs. Le test d'activation des basophiles (TAB) était positif à 40 % en présence du paracétamol (témoin négatif 2 % et témoin positif 93 %). Le TAB était négatif chez un patient témoin. En février 2019, le prick-test et l'IDR 10−2 étaient négatifs, mais l'IDR 10−1 était positive. Le test de provocation labiale au paracétamol a été bien toléré, mais la réintroduction de 1000 mg de paracétamol per os a provoqué une urticaire du tronc et une toux dans l'heure suivant la prise, sans autres symptômes systémiques. Une rapide résolution des symptômes a été obtenue sous antihistaminique intraveineux (dexchlorpheniramine). La tryptasémie était normale à 90 min. Nous avons diagnostiqué un rare cas d'hypersensibilité immédiate allergique au paracétamol et contre-indiqué la prise future de ce médicament chez ce patient. L'absence de signes de gravité peut expliquer la normalité de la tryptasémie lors de la dernière réaction. Nous avons donc mis en évidence un rare cas d'hypersensibilité immédiate allergique au paracétamol avec une IDR et un TAB positifs. Les TAB au paracétamol pourraient être intéressants, en complément des tests cutanés et de l'histoire clinique. Paracetamol is an analgesic used in the general population with few side-effects. However, rare anaphylactic reactions to this molecule have been reported, for which allergy workups were usually negative, in contrast with our case. A 19-year-old atopic male patient presented generalized urticaria with facial angioedema, dyspnea and loss of consciousness in November 2017, one hour after ingestion of a single dose of paracetamol 1000 mg. Symptoms improved within 30 min under oral antihistamine and prednisolone. Before and since this reaction, nonsteroidal anti-inflammatory drugs were tolerated by the patient. In October 2018, an immediate-reading prick-test (10 mg/mL) and an intradermal test (IDT) 10−2 (0.1 mg/mL) performed with paracetamol were negative. Oral intake of paracetamol 50 mg was well tolerated. A second series of skin tests was performed one month later and a paracetamol prick-test and IDT were positive. A basophil activation test (BAT) was positive at 40% in the presence of paracetamol (negative control 2% and positive control 93%). The BAT was negative in a control patient. In February 2019, a third series of skin tests with paracetamol was performed: a prick-test and IDT 10−2 were negative, but an IDT 10−1 (1 mg/mL) was positive. A labial challenge paracetamol test using paracetamol prick solution was well tolerated. One hour after an oral drug challenge with 1000 mg, the patient developed chest urticaria and cough with no other systemic symptoms. He recovered rapidly with intravenous antihistamine (dexchlorpheniramine). Serum tryptase was normal at 90 min. Allergic hypersensitivity to paracetamol was diagnosed and future intake of paracetamol was contraindicated. The absence of severe symptoms could account for the normal serum tryptase value during the last reaction. We report herein a rare case of immediate allergic hypersensitivity to paracetamol with a positive IDT and basophil activation test. BAT for paracetamol could be a reliable diagnostic tool in conjunction with skin tests and clinical history. [ABSTRACT FROM AUTHOR]

Furosemide is a widely used diuretic belonging to the family of loop diuretics, which is potent and rapid acting. It belongs to the chemical family of sulfonamides. Few cases [1,2] of hypersensitivity reactions to furosemide have been reported. We describe the case and the allergy work-up of a 43-year-old Caucasian woman with a history of an anaphylactic reaction after taking iv furosemide. Skin tests (ST) were performed as recommended, with prick (SPT) and intradermal test (IDT). The concentrations were experimental but verified in controls. The patient had experienced diffuse urticaria, respiratory distress and bradycardia at 40 bpm, 45 minutes after the injection of IV furosemide. Admitted to the emergency room she was treated with IV adrenaline, methylprednisolone hemisuccinate and oxygenotherapy (complete recovery after a few hours). No serum tryptase was collected. Prior to this episode, the patient had never taken furosemide. SPT with furosemide (neat concentration 10 mg/mL) was negative with a normal response to the histamine control and no response to the saline control. The IDT was positive at a concentration of 1:100. Three controls were all negative to the IDT at this concentration. IDT to furosemide at 1:1000 was also positive. Basal tryptase was 5.01 μg/L. We presented a case of anaphylaxis to furosemide, whose mechanism was proven to be IgE-mediated, based on the following arguments: severe clinical history, positive ST with negative controls at the same experimental concentrations. The allergological reasoning is the same, whether the external accountability for a certain drug is high or low. Cross reactivity with other sulfonamides should be further explored [3]. [ABSTRACT FROM AUTHOR]

Immediate-type hypersensitivity (ITH), mediated by IgE, to Malassezia pachydermatis is recognized in atopic dogs with recurrent yeast dermatitis and otitis externa (OE). Malassezia -associated OE commonly occurs in dogs without other signs of atopic dermatitis (AD). The aim of this study was to detect Malassezia -specific IgE in the sera of dogs with recurrent Malassezia OE without concurrent skin disease. Sera from healthy dogs were used for comparison. An FcεRIα-based ELISA was used to measure Malassezia -specific IgE. There was no significant difference between number of positive affected dogs (6/21, 29%) and number of positive unaffected dogs (15/86, 17%) ( P = 0.36). There was also no significant difference in the concentrations of Malassezia- specific IgE between the two groups ( P = 0.97). Malassezia- specific IgE did not distinguish between patient groups so, as with other canine allergens, serum IgE reactivity for Malassezia could not be used to differentiate between diseased and healthy patients. The presence of Malassezia -specific IgE in some of the affected dogs might indicate ITH to Malassezia in those dogs. Evaluation of ITH via intradermal test reactivity and response to allergen-specific immunotherapy might clarify the role of Malassezia -associated ITH in similarly affected dogs. [ABSTRACT FROM AUTHOR]

Canine atopic dermatitis (AD) is associated with increased levels of allergen-specific IgE due to hyper-sensitization to environmental allergens. Intradermal testing (IDT) and allergen-specific IgE serology testing are often used to determine the allergens which elicit an IgE response in animals with a diagnosis of AD. The objective of this study was to determine the effects of oclacitinib on IDT and allergen-specific IgE serology testing using a laboratory model of house-dust mite sensitized Beagle dogs. Twenty-four (24) normal, healthy purpose-bred Beagle dogs were sensitized to house dust mites (HDM, Dermatophagoides farinae) and randomly assigned to placebo-, oclacitinib- (0.4 mg/kg/dose PO), or prednisolone-treated (0.5 mg/kg/dose PO) groups. After 14 days of twice daily dosing, the effects of prednisolone and oclacitinib were compared to placebo using baseline and post-dose IDT and allergen-specific IgE serum measurements. Sensitized dogs had increased circulating HDM-specific IgE for at least two weeks post-sensitization. Prednisolone significantly inhibited the measurable sensitivity of IDT, while oclacitinib did not. Neither prednisolone nor oclacitinib imposed significant effects on allergen-specific IgE serum levels, suggesting oclacitinib may have potential to be used in dogs concurrently undergoing intradermal skin testing and/or allergen-specific IgE serology testing without interference with test results. • Oclacitinib given at 0.4 mg/kg/dose twice daily for 14 days neither inhibits allergen-specific IgE serology nor intradermal test responses when compared to placebo. • Prednisolone given at 0.5 mg/kg/dose twice daily for 14 days significantly reduces the intradermal testing response using serial dilutions of the sensitizing allergen. • Dogs with atopic dermatitis can remain on oclacitinib treatment for relief of clinical symptoms while undergoing serology or intradermal testing to identify allergens for avoidance or inclusion in allergen-specific immunotherapy. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) in goats ( Capra hircus ) is due to infection with members of the Mycobacterium tuberculosis complex (MTC), mainly Mycobacterium bovis and Mycobacterium caprae . We report a comparative experimental infection of goats with M. bovis , M. caprae and M. tuberculosis strains. We hypothesized that goats experimentally infected with different members of the MTC would display different clinical pictures. Three groups of goats were challenged with either M. bovis SB0134 (group 1, n = 5), M. caprae SB0157 (group 2, n = 5) and M. tuberculosis SIT58 (group 3, n = 4). The highest mean total lesion score was observed in M. bovis challenged goats (mean 15.2, range 9–19), followed by those challenged with M. caprae (10.8, 2–23). The lowest score was recorded in goats challenged with M. tuberculosis (3, 1–6). Culture results coincided with the lesion scores in yielding more positive pools (7/15) in M. bovis challenged goats. By contrast, only three pools were positive from goats challenged M. tuberculosis (3/12) and with M. caprae (3/15), respectively. Differences in the performance of the intradermal and gamma-interferon (IFN-γ) tests depending of the group were observed since all goats from group 1 were diagnosed using intradermal test and these goats reacted earlier to the IFN-γ assay in comparison to the other groups. This study confirmed that goats experimentally infected with different members of the MTC display different clinical pictures and this fact may have implications for MTC maintenance and bacterial shedding. [ABSTRACT FROM AUTHOR]

Insect bite hypersensitivity (IBH) is an allergic dermatitis of horses caused by bites of midges ( Culicoides spp.). IgE-mediated reactions are often involved in the pathogenesis of this disease. IBH does not occur in Iceland due to the absence of Culicoides , but it occurs with a high frequency in Icelandic horses exported to mainland Europe, where Culicoides are present. We hypothesize that immunization with the Culicoides allergens before export could reduce the incidence of IBH in exported Icelandic horses. The aim of the present study was therefore to compare intradermal and intralymphatic vaccination using four purified recombinant allergens, in combination with a Th1 focusing adjuvant. Twelve horses were vaccinated three times with 10 μg of each of the four recombinant Culicoides nubeculosus allergens. Six horses were injected intralymphatically, three with and three without IC31 ® , and six were injected intradermally, in the presence or absence of IC31 ® . Antibody responses were measured by immunoblots and ELISA, potential sensitization in a sulfidoleukotriene release test and an intradermal test, cytokine and FoxP3 expression with real time PCR following in vitro stimulation of PBMC. Immunization with the r-allergens induced a significant increase in levels of r-allergen-specific IgG1, IgG1/3, IgG4/7, IgG5 and IgG(T). Application of the r-allergens in IC31 ® adjuvant resulted in a significantly higher IgG1, IgG1/3, IgG4/7 allergen-specific response. Intralymphatic injection was slightly more efficient than intradermal injection, but the difference did not reach significance. Testing of the blocking activity of the sera from the horses immunized intralymphatically with IC31 ® showed that the generated IgG antibodies were able to partly block binding of serum IgE from an IBH-affected horse to these r-allergens. Furthermore, IgG antibodies bound to protein bands on blots of C. nubeculosus salivary gland extract. No allergen-specific IgE was induced and there was no indication of induction of IgE-mediated reactions, as horses neither responded to Culicoides extract stimulation in a sulfidoleukotriene release test, nor developed a relevant immediate hypersensitivity reaction to the recombinant allergens in skin test. IL-4 expression was significantly higher in horses vaccinated intralymphatically without IC31 ® , as compared to horses intradermally vaccinated with IC31 ® . Both routes gave higher IL-10 expression with IC31 ® . Both intralymphatic and intradermal vaccination of horses with recombinant allergens in IC31 ® adjuvant induced an immune response without adverse effects and without IgE production. The horses were not sensitized and produced IgG that could inhibit allergen-specific IgE binding. We therefore conclude that both the injection routes and the IC31 ® adjuvant are strong candidates for further development of immunoprophylaxis and therapy in horses. [ABSTRACT FROM AUTHOR]

Résumé Les toxidermies correspondent aux effets indésirables médicamenteux à expression cutanéo-muqueuse. Elles présentent une grande variabilité sémiologique, non spécifique de l’étiologie médicamenteuse. L’exanthème maculo-papuleux est la présentation la plus fréquente suivie de l’urticaire et des vascularites. L’érythème pigmenté fixe est une toxidermie plus rare dans les pays occidentaux. La fréquence des formes sévères (décès, séquelles graves) est de 2 % : toxidermies bulleuses (syndrome de Stevens-Johnson, nécrolyse épidermique toxique ou syndrome de Lyell), DRESS ( drug reaction with eosinophilia and systemic symptoms ou syndrome d’hypersensibilité), pustulose exanthématique aiguë généralisée (PEAG). Elles doivent être rapidement identifiées afin d’orienter la prise en charge. Les principaux facteurs de risque des toxidermies sont l’immunodépression, les pathologies auto-immunes ainsi que certains génotypes du système HLA dans les toxidermies bulleuses et le syndrome d’hypersensibilité. Toutes les classes médicamenteuses peuvent être à l’origine de toxidermies, en particulier les antibactériens, les anticonvulsivants, les antinéoplasiques, les anti-inflammatoires non stéroïdiens, l’allopurinol et les produits de contraste. Les mécanismes physiopathologiques comprennent les réactions immunologiques/immuno-allergiques immédiates ou retardées, classiquement non dose-dépendantes, et les mécanismes toxiques/pharmacologiques dose-dépendants ou temps-dépendants, incluant les réactions dites « paradoxales ». En cas de toxidermie de mécanisme immuno-allergique, un bilan allergologique est possible pour préciser l’imputabilité du/des médicament(s) suspect(s) avec une sensibilité variable selon les médicaments et le type de toxidermie. Il comprend les tests épicutanés (ou patch tests) et les tests intradermiques (prick test et intradermoréactions). Cependant aucun test in vivo ou in vitro ne permet d’affirmer la responsabilité médicamenteuse devant une toxidermie. Le diagnostic repose donc sur une démarche d’imputabilité prenant en compte la présentation clinique, la chronologie des événements et des prises médicamenteuses et l’élimination des diagnostics différentiels. Cette démarche diagnostique, complétée par une recherche bibliographique, permet dans la majeure partie des cas d’établir un lien de causalité entre le(s) médicament(s) et l’éruption cutanée. Une déclaration en pharmacovigilance est indispensable en cas de toxidermie grave quel que soit le médicament impliqué, et dans tous les cas s’il s’agit d’un médicament récent ou non classiquement associé à un risque cutané. Cutaneous adverse drug reactions (CADR) represent a heterogeneous field including various clinical patterns without specific features suggesting drug causality. Exanthematous eruptions, urticaria and vasculitis are the most common forms of CADR. Fixed eruption is uncommon in western countries. Serious reactions (fatal outcome, sequelae) represent 2% of CADR: bullous reactions (Stevens-Johnson syndrome, toxic epidermal necrolysis), DRESS (drug reaction with eosinophilia and systemic symptoms or drug-induced hypersensitivity syndrome) and acute generalized exanthematous pustulosis (AGEP). These forms must be quickly diagnosed to guide their management. The main risk factors are immunosuppression, autoimmunity and some HLA alleles in bullous reactions and DRESS. Most systemic drugs may induce cutaneous adverse reactions, especially antibiotics, anticonvulsivants, antineoplastic drugs, non-steroidal anti-inflammatory drugs, allopurinol and contrast media. Pathogenesis includes immediate or delayed immunologic mechanism, usually not related to dose, and pharmacologic/toxic mechanism, commonly dose-dependent or time-dependent. In case of immunologic mechanism, allergologic exploration is possible to clarify drug causality, with a variable sensitivity according to the drug and to the CADR type. It includes epicutaneous patch testing, prick test and intradermal test. However, no in vivo or in vitro test can confirm the drug causality. To determine the cause of the eruption, a logical approach based on clinical characteristics, chronologic factors and elimination of differential diagnosis is required, completed with a literature search. A reporting to pharmacovigilance network is essential in case of a serious CADR whatever the suspected drug and in any case if the involved drug is a newly marketed one or unusually related to cutaneous reactions. [ABSTRACT FROM AUTHOR]

Continual tailoring of control programmes of endemic pathogens during long-term eradication campaigns requires detailed analysis of surveillance data to inform evidence-based policy. Bovine tuberculosis is a disease where long-term control and eradication programs are in train in several countries. The primary diagnostic tool, the intradermal tuberculin test, used to identify infected animals can be interpreted using different criteria and cut-offs, facilitating flexibility in its use as a basis to inform interventions. We investigated the comparative risk of animals failing a single intradermal comparative tuberculin test (SICTT) based on their previous tuberculin test result following a higher risk test-type (reactor retest of an infected herd). The study was a retrospective cohort design, and the primary exposure was the test status following a reactor retest classified as mutually exclusive categories based on bovine and avian tuberculin reactions: standard interpretation inconclusive (Sdi), severe interpretation inconclusive (Svi), single intradermal test (SIT) reactors (SITr), SIT inconclusive (SITi), avian tuberculin reactors (Ar), and test negative animals. Random effects multivariable logistic regression was used to investigate future risk. Cross-validation and downscaling was used to explore model performance. Alternate models with differing outcome test types were also explored. The models were trained on 844,207 observations from June 2018 to June 2021. Sdi, Svi, SITr, SITi and Ar were associated with the following odds ratios 12.242 (95 %CIs: 5.236–28.625; p < 0.001), 4.101 (95 %CIs: 3.423–4.913; p < 0.001), 2.503 (95 %CIs: 1.878–3.338; p < 0.001), 1.741 (95 %CIs: 1.195–2.538; p = 0.004) and 1.065 (95 %CIs: 0.833–1.361; p = 0.616) for failing the next test, respectively. High model performance was achieved with inclusion of random effects for both training and test evaluation datasets (AUC: 0.94; Balanced accuracy: 0.84), but fixed-effects only predictions exhibited moderate performance (AUC: 0.70; Balanced accuracy: 0.69). This reflects that 55 % of the risk of test failure relates to between herd heterogeneity based on intra-class correlation, while controlling for fixed effects. Other factors that were associated with increasing risk included age (older cohorts were at greater risk than the youngest cohort), breakdown history of the herd (greater number of breakdowns prior to the study period), and the time between exposure test and outcome test. These results provide further evidence to inform risk-based management policies for TB, including the removal of Sdi animals in higher risk situations, supplementary testing of cattle based on tuberculin responses and the provision of risk management advice to herd owners. The results characterise the future animal-level risk posed by Svis and suggest this risk may require policy led interventions. • Tuberculin test results can provide rich information on skin response sizes. • Such data are often ignored beyond standard binary test cut-offs. • Future risk of test failure based on 'exposure' to six response types was tested. • All defined reactor types based on bovine reactions were at increased risk. • Severe inconclusive reactors may be considered for future policy interventions. [ABSTRACT FROM AUTHOR]

Abstract: Tuberculosis (TB) in llamas and alpacas has gained importance in recent years since they are imported into the European Union mainly for serving as pets and for production of natural fibre. The intradermal tuberculin test has been widely used for diagnosis of TB in these species showing lack of sensitivity (Se) although little information has been previously reported evaluating the effect on its performance of different PPD inoculation sites and time of readings. Moreover, different cost-effective serological assays have been developed in the recent years for TB diagnosis in camelids obtaining a variety of results and, for this reason, new assays still being developed. The main objectives of this study were: (1) to evaluate the performance of the intradermal tuberculin test using different inoculation sites (axillary, prescapular and cervical) and times of reading (72 and 120h) and (2) to test a novel serological assay based on MPB83 antigen in a Mycobacterium bovis naturally infected alpaca herd in Spain. In regards to skin test, single intradermal tuberculin (SIT) test at the prescapular site and reading at 72h showed the highest proportion of test-positive-culture positive animals among all culture positive animals (T+/C+), ranging from 53.8% (95% CI, 37.2–69.9) to 80% (95% CI, 44.4–97.5) using a more stringent interpretation than typically prescribed although, in general, low T+/C+ was achieved using both SIT and single comparative intradermal tuberculin (SCIT) tests alone. T+/C+ of the serological assay increased using samples collected 15–30 days after PPD injection [76.9% (95% CI, 60.7–88.9) – 100% (95% CI, 69.2–100)]. The best results of T+/C+ were obtained applying in parallel the most sensitive SIT test and serology using samples collected 15–30 days after PPD inoculation [90% (95% CI, 55.5–99.7)-100% (95% CI, 69.2–100)]. Therefore implementation of serology in parallel with the most sensitive skin test could maximize the detection of infected animals. [Copyright &y& Elsevier]

Abstract: Insect bite hypersensitivity (IBH) is an IgE-mediated (Type I) hypersensitivity reaction induced by allergens from biting midges of the Culicoides spp. The aim of the present study was to identify, clone and express recombinant allergens from C. obsoletus, the main species found feeding on horses in the Netherlands, by sequence homology searches on the C. obsoletus specific RNA database, with previously described allergens from C. nubeculosus and C. sonorensis. BLAST searches with these described allergens resulted in similarity hits with 7 genes coding for C. obsoletus allergens. These allergens were expressed as hexahistidine tagged recombinant proteins in E. coli. Allergens were termed Cul o 1–Cul o 7. A maltase (Cul o 1) plus Cul s 1 (maltase of C. sonorensis) were additionally expressed in insect cells using the baculovirus expression system to compare homologous allergens from different species produced with different expression systems in diagnostic in vitro and in vivo tests. We demonstrate that IBH affected horses in the Netherlands show higher IgE levels to Cul o 1 than to Cul s 1, as determined by an IgE ELISA. Furthermore, we show that Cul o 1 produced in E. coli is at least as suitable for in vitro diagnosis of IBH affected horses as Cul o 1 produced in the baculovirus/insect cell expression system. The resulting proteins were evaluated for their ability to discriminate IBH affected and healthy horses by ELISA and intradermal testing. The frequency of positive test results by ELISA within IBH affected horses ranged from 38% to 67% for the different allergens. When results of IgE-binding to Cul o 1–Cul o 7 were combined the test had a sensitivity of 92% and specificity of 85%. The capability of the allergens to induce Type I hypersensitivity reaction in IBH affected horses was demonstrated by an intradermal test. The results show that E. coli expressed recombinant allergens from C. obsoletus are valuable tools to determine the allergen specific sensitisation profile (component resolved diagnosis) in horses with IBH in countries were C. obsoletus is the most abundant species and may facilitate in the development of future immunotherapy. [Copyright &y& Elsevier]

Summary: A Mycobacterium bovis strain deleted in mce2A and mce2B genes (M. bovis Δmce2) was tested as an experimental vaccine in cattle challenged with a virulent M. bovis strain. Three-and-a-half-month old calves (n = 5 to 6 per group) were vaccinated and challenged with a virulent strain of M. bovis by the intratracheal route 9 weeks after vaccination. A non-vaccinated group and a group vaccinated with BCG were included as controls. Blood samples were collected to measure IFN-γ by an interferon-gamma release assay (IGRA), cytometry and cytokine responses of bovine purified protein derivative (PPD) restimulated peripheral blood mononuclear cells (PBMCs). The IGRA test showed IFN-γ values similar to pre-vaccination except for the animals vaccinated with M. bovis Δmce2, where a significant increase was observed at 30 days post-vaccination. The expression of IL-2R on CD4+ cells in response to PPD from the animals vaccinated with Δmce2 increased at 15 days post-vaccination compared to cells from non-vaccinated group. Vaccination of cattle with M. bovis Δmce2 induced the highest (P < 0.05) expression of IFN-γ and IL-17 mRNA upon PPD stimulation of PBMCs compared to vaccination with BCG or that for the non-vaccinated group. There was a weak positive correlation between the production of these proinflammatory cytokines post-vaccination and reduced pathology scores post-challenge. The animals were euthanized and necropsied 100 days after challenge. The group vaccinated with M. bovis Δmce2 displayed a significantly lower histopathological score for lesions in lungs and pulmonary lymph nodes than for the other groups (P < 0.05). A marked positive reaction to tuberculin intradermal test was observed post-vaccination in animals vaccinated with M. bovis Δmce2 compared to those vaccinated with BCG or the non-vaccinated group. In contrast, after challenge, non-vaccinated animals had greater skin test responses than the vaccinated animals. In summary, M. bovis Δmce2 is a promising vaccine candidate to control M. bovis pathogenesis in cattle. [Copyright &y& Elsevier]

Abstract: Paragonimiasis has previously been reported in the Southwest Provinces of China, including Chongqing and Sichuan. The construction of Three Gorges Dam, which was begun in 1994, has resulted in substantial changes to the depth and the flow pattern of the Yangtze River. To investigate epidemiology of the paragonimiasis, 724 people aged 2–49years were selected and examined for Paragonimus infection by intradermal test (IDT), and enzyme-linked immunosorbent assay (ELISA). A survey of eating habits was conducted face-to-face in 5 endemic counties. Freshwater crabs were collected from seven sites in the 5 counties to determine the rate of infection with Paragonimus metacercariae. Additionally, we analyzed 213 case reports from the region dated 2006 to 2009. According to the survey results, the prevalence of Paragonimus skrjabini infection in freshwater crab was 39.65%, and the human infection rates detected by IDT and ELISA were 14.36% (104/724) and 7.46% (54/724), respectively. These results show that the Three Gorges Reservoir is still an area of high paragonimiasis endemicity. The water level of the Yangtze River has risen to 175m, and the area of water coverage in the region has increased to 1084km2. These conditions will favor the growth and expansion of the freshwater crab population. [Copyright &y& Elsevier]

Abstract: Fallow deer (Dama dama) are widely distributed as natural or naturalised populations, as well as in game parks and deer farms. We used 157 fallow deer sampled in populations considered to be Mycobacterium tuberculosis complex (MTC) free and 73 Mycobacterium bovis-infected fallow deer confirmed postmortem by culture to evaluate the diagnostic performance of two tests for the detection of anti-mycobacterial antibodies: the dual path platform (DPP) VetTB assay and the bovine purified protein derivative (bPPD) ELISA. We also compared their sensitivity with that of the skin test, analyzed the effect of haemolysis degree on the antibody detection and described the relationship between the test readings and presence/absence of gross tuberculosis (TB) compatible lesions. Sensitivity of bPPD ELISA was 51% at a specificity of 96%. Depending on the cut-off value selected, the sensitivity of DPP VetTB ranged from 62 to 71%, while its specificity was 88–95%. In the subgroup of M. bovis-infected deer for which the skin test data were available (33 of 73); this method detected 76% of culture-positive animals, although the specificity of the intradermal test was not determined in this study. When the DPP VetTB and skin test data were combined, the resulting sensitivity obtained in this sub-group of M. bovis-infected deer increased to 97%. Gross pathology identified TB compatible lesions (TBL) in 89% culture-confirmed fallow deer. The infected animals with visible lesions had significantly higher readings in the DPP VetTB, but not in the bPPD ELISA. Only high levels of haemolysis decreased antibody test sensitivity and this effect was more evident for the bPPD ELISA. The results allowed inferring a number of management recommendations for rapid detection of MTC infection in live fallow deer and in surveys on hunter-harvested cervids. [Copyright &y& Elsevier]

Background: Vital dyes are widely used for lymphatic mapping and sentinel lymph node biopsy in patients with malignant tumors, and reports of anaphylactic reactions are becoming more frequent. Objective: Our aims were to describe specific clinical features of hypersensitivity reactions to Patent Blue (Guerbet, Roissy, France), results of the allergy workup, and their consequences for patient management. Methods: We report a series of 14 clinical cases of dye-induced anaphylaxis recorded between 2004 and 2006 in 4 member centers of a network of French allergoanesthesia outpatient clinics. Results: Reactions appeared to be relatively severe (6/14 grade III reactions). An average 30 ± 6–minute delay was observed between dye injection and symptom onset. In 9 (65%) patients reactions were sustained for several hours, requiring prolonged continuous epinephrine infusion and transfer to an intensive care unit. Prick test results were positive in 8 patients. In 5 patients prick test results were negative, whereas intradermal test results were positive. Conclusion: Anesthesiologists and allergologists must be aware of this specific risk and of the clinical characteristics of these reactions, which are usually delayed and long lasting. [Copyright &y& Elsevier]

Abstract: Parasitological techniques (stool examination and/or urine filtration) are still the diagnosis of choice in national schistosomiasis control programmes the world over. However, the success of current control efforts, mainly due to the large-scale administration of praziquantel, emphasises the need for a more sensitive approach. In addition, microscopy is labour-intensive and time-consuming, while declining compliance rates after repeated chemotherapy make good coverage for the long-term increasingly problematic. China''s success in the control of schistosomiasis is contributing to an enhanced understanding of the need for better and more sensitive screening methods. Immunodiagnostic techniques have a high sensitivity, are easy to perform and are an excellent epidemiological tool for the screening of target populations in schistosome-endemic areas. These assays are also useful for the surveillance of cure after chemotherapy, and for periodic control of transmission of the infection after it has been eliminated in an area. A succinct historical background of using immunodiagnosis for schistosomiasis japonica in China is given, together with a review and evaluation of the relative efficacy of the main techniques applied, i.e. the intradermal test, the circumoval precipitation test, the indirect hemagglutination assay, the dye dipstick immunoassay and different kinds of enzyme-linked immunosorbent assay applications. The important role of immunodiagnosis in the screening for schistosomiasis in China is discussed. [Copyright &y& Elsevier]

Background: In addition to immediate reactions, late adverse reactions to iodinated contrast media (ICM) were reported in 2% to 5% of patients exposed to ICM and, as a consequence, have recently gained more attention. A few well-documented case reports postulate a hypersensitivity mechanism. Objective: The aim of this study is to demonstrate a T cell–mediated mechanism to the ICM by using in vitro and ex vivo tests. Methods: We analyzed 12 patients with 13 adverse ICM reactions, 9 of whom were women. Clinical history suggested an immune reaction to ICM. Skin tests (skin prick, intradermal, and patch tests) were performed with various ICM and read after 15 minutes and 24 and 48 hours. Skin biopsy specimens of positive test sites of 11 patients were evaluated by means of immunohistology. T-cell reactivity to ICM in vitro was analyzed with lymphocyte activation tests. Results: Seven patients showed generalized maculopapular eruptions, one of them with fever; 4 had a so-called drug hypersensitivity syndrome with exanthema, eosinophilia, and fever; 1 had maculopapular eruptions and fever; 1 had late-onset urticaria with loss of consciousness; and 1 had facial edema and respiratory distress. An immune reaction to ICM was inferred from positive skin prick test (2 patients), positive patch test (10 patients), and positive intradermal test (9 patients) at 24 and 48 hours. Skin biopsy specimens revealed a T-cell infiltrate in the dermis with predominantly CD4+ T cells in 8 patients, CD8+ T cells in 1 patient, and equal numbers in 1 patient. Cross-sensitivities to several ICM were observed (9/12). Other drug allergies were noted in 6 of the 12 patients. Conclusions: Delayed reactions to ICM are most likely caused by immune reactions to these drugs and can elicit different clinical features. The involvement of T cells is suggested by positive skin test, as well as positive proliferative responses, to the drugs in vitro. A high degree of cross-reactivity with other than the eliciting ICM was observed. Moreover, 50% of these patients reported another drug hypersensitivity, suggesting a predisposition to immune reactivity in some patients. [Copyright &y& Elsevier]

Cats with spontaneously occurring atopic dermatitis have clinical and immunocytochemical characteristics compatible with these in humans with atopic dermatitis (AD). The atopy patch test (APT) has proven to be a valuable tool in elucidating the disease process in humans. Additionally, the APT is very specific and bypasses the problem of conflicting results due to differences in chronicity of lesions of AD patients.We adapted the APT for use in cats to explore the suitability of the APT as a tool to study the onset of allergic inflammation in cats with atopic dermatitis.APT were performed in AD cats (n = 6) and healthy cats (n = 10). All cats were patch tested with two allergens in three different dilutions and a diluent control. The allergens for the APT were selected from positive intradermal test and /or prick test results and consisted of: Dermatophagoides farinae, D. pteronyssinus, Tyrophagus putrescentiae, and a grass pollen mixture. APT were read after 10, 24 and 48 h, and punch biopsies for immunohistochemical evaluation were collected at these time points.Macroscopically positive APT reactions were observed in three out of six cats at 24 and/or 48 h with allergen concentrations of 25 000 and 100 000 NU/ml. Reactions were not observed at negative control sites and neither in control animals.A significantly increased number of IL-4+, CD4+, CD3+, MHC class II+ and CD1a+ cells was found in one AD cat with positive APT reactions. Five out of six AD cats had significantly increased IL-4+ T cell numbers at 24 and/or 48 h.Our data indicate that in cats, macroscopically positive patch test reactions can be induced, which have a cellular infiltrate similar to that in lesional skin. We found a high specificity and a macroscopically positive APT reaction in half of the cats, which is similar to what is seen in humans. Hence, the APT in cats might be a useful tool in studying the immunopathogenesis of feline atopic dermatitis. [Copyright &y& Elsevier]

In this study, we investigated the development of clinical disease and immune responses in the development of an experimental model of flea allergy dermatitis. Dogs were randomly divided into four treatment groups and were infested with fleas on two different feeding schedules (continuous and episodic). Group 1 consisted of four non-exposed dogs (negative controls) and Group 2 consisted of six dogs exposed to fleas continually. Groups 3 and 4 consisted of 14 dogs each that were exposed to fleas on an episodic schedule (two consecutive days every other week for 12 weeks). Group 4 also received intraperitoneal injections of a low dose of lectin (ricin) with immunomodulatory properties. The purpose of Group 4 was to investigate the effects of ricin on enhancing the development of clinical signs, flea antigen-specific IgE levels and altering the number of CD4+ and CD8+ T cell subsets in peripheral blood. Clinical signs developed in all flea exposed dogs, however, the dermatology lesion scores were less and shorter in duration for continuously exposed dogs compared to episodic exposed dogs, independent of ricin treatment. Lesion development was concentrated in the flea triangle and consisted principally of erythema, followed by alopecia, excoriation, papules, and crusts. CD4+ and CD8+ lymphocyte subsets or IgE levels were not altered by ricin treatment. Flea antigen-specific IgE values were highest in dogs exposed to fleas on a continuous basis compared to those episodically exposed. A greater percentage of clinical responder dogs with negative flea-specific IgE titers or negative intradermal test (IDT) were present in the episodic exposure groups than in the continuous exposure group. IgE titers corresponded slightly better with clinical responders than the IDT. The agreement between the IgE titers and IDT was good (weighted κ=0.67). Histopathology of skin samples were consistent with a Type I hypersensitivity. In conclusion, we were able to develop a model of flea allergy dermatitis by experimentally exposing dogs to fleas on an episodic and continuous feeding schedule. In this study, continuously exposed dogs did not develop immunotolerance, and ricin did not enhance the development of FAD. [Copyright &y& Elsevier]

To identify optimal method(s) for certification and subsequent monitoring of Mycobacterium avium subsp. paratuberculosis (Map)-unsuspected herds, certification-and-monitoring schemes were studied using a stochastic simulation model (“JohneSSim”). JohneSSim simulated the within-herd transmission and economic aspects of Map in closed Dutch dairy herds. The model was validated with field observations on Map-unsuspected herds. The current Dutch certification-and-monitoring schemes were compared with 11 alternative schemes in which individual and pooled fecal culture, ELISA, Johnin-intradermal test and γ-IFN ELISA were used, varying the test frequency, tested age group and number of tested animals.On reaching the ‘Map-free’ status with the standard certification scheme, 11% of the simulated herds were not truly Map-free. Therefore, the designation ‘Map-free’ should be changed into, for instance, ‘low-risk Map’. In the most-attractive alternative certification scheme, the ‘Map-free’ status was reached after four herd examinations (at 2-year intervals) consisting of serial testing of all cattle≥2 years of age with a pooled fecal culture and individual fecal culture of positive pools. This scheme resulted in lower total and annual discounted costs and a lower animal-level prevalence at reaching the ‘Map-free’ status compared to the standard scheme, assuming that there was no new introduction of the infection.Schemes to monitor the ‘Map-free’ status were compared, assuming that this status was reached with the standard certification scheme. In comparison to the standard monitoring scheme, none of the alternative monitoring schemes resulted in both a lower animal-level prevalence of undetected pre-existing Map infections in closed herds, and lower median annual discounted costs.Results of the model were very sensitive to the assumed sensitivity of the fecal culture test and to management measures that prevent within-herd transmission of Map. If these preventive measures were taken, the probability of undetected Map infections in closed ‘Map-free’ herds was decreased substantially. [Copyright &y& Elsevier]

Tuberculosis (TB) in cattle remains a major zoonotic and economic problem in many countries. Since the standard diagnostic assay, the intradermal test (IDT) with bovine PPD tuberculin, has less than optimal accuracy in all situations, other diagnostic methods such as serological assays have been investigated. Because of fundamental concerns for the low sensitivity and specificity of previous ELISA protocols, a profiling ELISA with nine purified, recombinant proteins of TB complex mycobacteria, was employed on samples from four groups of cattle: (a) naturally Mycobacterium avium-exposed and experimentally Mycobacterium bovis-infected, (b) officially-certified TB-free herds, (c) exposed to M. bovis in two field TB outbreaks and scored as bovine reactors in the γ-IFN assay for bovine TB, (d) paratuberculosis (para TB)-infected. The described ELISA proved to be highly specific. In fact, the antibody (Ab) response could be consistently detected in 3 out of 3 endotracheally-infected calves and in 1 out of 3 contact-infected calves. There was also a very low prevalence of low-titered, non-specific Ab responses in paraTB-infected animals. As for the animals exposed to field TB outbreaks, 16 out of 28 γ-IFN positive cattle were also Ab-positive; importantly, 7 out of 12 γ-IFN positive, IDT-negative cattle showed Ab responses to TB proteins. In general, the profile of the Ab response varied among animals; the reaction to single recombinant antigens was sometimes transient and fluctuating, whereas the panel of antigens on the whole was indeed more effective in Ab detection. [Copyright &y& Elsevier]

Bovine tuberculin purified protein derivative (bPPD) is used as an intradermal test (IT) reagent to detect bovine tuberculosis (bTB) in most countries. Identification of bPPD proteins is critical to understanding the immunological reaction of IT at the molecular level. While bPPD from the United Kingdom (UK) and Brazil (BR) have been recently defined at the proteomic level, bPPD from the Republic of Korea (KR) has not yet been analyzed. Here, bPPD KR proteome was examined for the first time. In total, 271 proteins were identified, including Mycobacterium bovis-specific proteins Mb0854c and Mb2898, and 42 known T cell antigens. On comparing with proteomes of bPPD UK and BR, 33 proteins were found to be common among all three bPPDs, of which 15 proteins were T cell antigens. M. bovis-specific antigens with T cell activity in bPPD may be novel candidates for use as alternatives to currently available bPPD in diagnostics. [ABSTRACT FROM AUTHOR]

Potential triggers for equine asthma are allergens from hay and straw dusts, mold spores and storage mites. The contribution of these environmental trigger factors to equine asthma is still largely uncertain. The aim of this study was to compare results of four allergy tests from healthy and asthma-affected horses, and to evaluate the clinical relevance of allergens tested positive via specific inhalation provocation test. Fifteen horses were classified using a clinical scoring system as asthmatic (n = 9) or control (n = 6). Four different allergy tests (functional in vitro test, intradermal test, Fc-epsilon receptor test, and ELISA for allergen-specific IgE) were compared. A histamine inhalation provocation test as positive control was performed in all horses and the interpleural pressure was measured. In addition, two individual allergens were chosen for the allergen inhalation provocation test based on the results of the allergy tests and inhaled in increasing concentrations, until signs of dyspnea occurred. None of the four allergy tests could differentiate reliably between controls and asthma-affected horses. There was no agreement among the results of the four allergy tests. The interpleural pressure results showed a large individual variability. A clear positive reaction on the allergen inhalation provocation test was only detected in two asthma-affected horses 6 hours after allergen inhalation with Aspergillus fumigatus and Cladosporium herbarum. In most cases a purely type I immediate reaction is unlikely to be involved in causing the clinical signs of equine asthma. Because of a delayed reaction after allergen provocation in two horses, the involvement of cell-mediated type III or IV hypersensitivity may be possible. As all allergy tests used in this study can only detect IgE-mediated hypersensitivity, these tests are probably not suitable for an etiological diagnosis of equine asthma. [ABSTRACT FROM AUTHOR]

Abstract: The goal of the present work was to develop reagents with potential for tuberculosis diagnosis. Genetic sequences of Mycobacterium tuberculosis secretion antigens were amplified by PCR, cloned into the Gateway® system, and expressed in Escherichia coli. The recombinant M. tuberculosis proteins were purified by metal affinity chromatography and preparative gel SDS-PAGE electrophoresis followed by electroelution and removal of endotoxins using Triton X-114. In total, seven recombinant proteins were obtained (ESAT-6, CFP10, TB10.3, TB10.4, MTSP11, MPT70, and MPT83). Delayed hypersensitivity reactions (DHR) was evaluated in Cavia porcellus and compared to the response using a standard purified protein derivative (PPD). All seven recombinant proteins produced a positive induration reaction in an intradermal test in guinea pigs previously sensitized with M. tuberculosis. When applied together, at a concentration of each recombinant protein 0.04mg/mL, the intradermoreaction in C. porcellus was significantly higher than that obtained by standard PPD (p-value=0.00386). [Copyright &y& Elsevier]

Background: In order to identify the sentinel lymph node (SLN) in melanoma patients intradermal injection of a radiocolloid tracer and a blue dye are commonly used. Life-threatening side effects such as allergic reactions to the injected dye have been described. We report 3 cases with systemic allergic reactions.Methods: Three patients suffering from systemic reactions such as hypotension and rash during SLN biopsy were tested for sensitisation against patent blue and methylene blue with skin prick, scratch, and intradermal test.Results: All 3 patients showed positive skin tests to patent blue confirming allergic reaction to the injected dye. In addition, all patients showed positive skin tests with methylene blue indicating immunologic cross-reactivity between patent blue and methylene blue.Conclusions: Although allergic reactions to blue dye during SLN biopsy are rare, they may be life threatening. It is important that the attending anesthetist is aware of this. [ABSTRACT FROM AUTHOR]

Venom immunotherapy is the only curative intervention for subjects with Hymenoptera venom allergy who suffering systemic reactions upon bee or wasp stings. Venom immunotherapy can restore normal immunity against venom allergens, as well as providing to allergic subjects a lifetime tolerance against venoms. Nevertheless, it is necessary using safety assays to monitoring the development of tolerance in the VIT protocols to avoid fatal anaphylactic reactions. The purpose of this study was to assess the modifications in several markers of tolerance induction in subjects with Hymenoptera venom allergy undergoing immunotherapy. The studies were performed at baseline time and after six month of VIT. Intradermal skin tests, basophil activation tests, specific IgE levels; and the T-cell markers (IL-4 and IFN-γ producing cells; and expression of the surface activation markers CD40L and CTLA-4) were assayed. At six month of imunotherapy all parameters studied had significant alterations. All decreased, except the IFN-γ producing cells. In addition, modifications in intradermal skin test showed a significant correlation with both, CD40L expression on CD4 T lymphocytes ( p = 0.043) and IL-4 producing T lymphocytes ( p = 0.012). Neither basophil activation test nor serum levels of sIgE demonstrated any correlation with the immunological parameters studied nor among them. These results suggest that both IL-4 production and CD40L expression could be two good indicators of the beneficial effects of venom immunotherapy which translate into skin tests. [ABSTRACT FROM AUTHOR]

Mycobacterium bovis causes 3.1% of human tuberculosis cases, as described by the World Health Organisation. In cattle, this organism causes bovine tuberculosis (BTB) which can have a prevalence of up to 39.5% in some developing countries. In developed countries, although the prevalence of BTB has been reduced through eradication programmes, complete eradication has in some cases proved elusive, with prevalences in cattle of 0.5% in the Republic of Ireland and of 4.3% in the UK. As the tuberculous granuloma is the fundamental lesion that reflects the pathogenesis, immune control and progression of BTB, we aimed to develop an in vitro model of the early-stage bovine tuberculous granuloma, in order to model the early stages of BTB, while also reducing the use of experimentally infected animals. In vitro models of human and ovine mycobacterial granulomas have previously been developed; however, so far, there is no model for the BTB granuloma. As the disease in cattle differs in a number of ways from that in other species, we consider this to be a significant gap in the tools available to study the pathogenesis of BTB. By combining bovine monocyte-derived macrophages infected with M. bovis -BCG and autologous lymphocytes we have developed an early-stage tuberculous bovine granuloma model. In the model, 3D cell aggregations formed a spherical-shape that grew for up to 11 days post-infection. This bovine tuberculous granuloma model can aid in the study of such lesion development, and in comparative studies of pathogenesis, such as, for example, the question of mycobacterial latency in bovine tuberculosis. [ABSTRACT FROM AUTHOR]

Summary: Objective: To present the clinicopathological features and the practice of diagnosing infectious cutaneous granulomas in Egypt. Methods: This study included all cases diagnosed with infectious cutaneous granuloma during the period 2004–2010 at Al-Hussein University Hospital, Cairo. Clinical and histological features were recorded, along with the positivity rate (PR) for each diagnostic method. Results: This study included 233 cases (150 males and 83 females) with a mean age of 47 years. Three groups of infection were recorded: bacterial infections (73.8% Mycobacterium and 3.9% non-Mycobacterium), parasitic infestations (16.7%), and deep fungal infections (5.6%). Tuberculosis cases formed the largest granuloma group (40.8%), followed by leprosy (31.7%) and leishmaniasis (15.9%). A total of 36 cases were diagnosed by direct smear (PR 15.5%), 61 cases by skin biopsy (PR 31.0%), 84 cases by intradermal test (PR 63.6%), 26 cases by serological tests (PR 60.5%), 18 cases by tissue culture (PR 69.2%), and eight cases by PCR (PR 100%). Conclusions: Mycobacterial infections constitute the most common infectious cutaneous granulomas among Egyptians. Routine methods such as direct smear, skin biopsy, and intradermal tests remain the most commonly applied and economical methods for diagnosis in developing countries, although specific methods such as tissue culture and PCR have higher positivity rates in the diagnosis. [ABSTRACT FROM AUTHOR]

Abstract: The objective of this study was to compare and analyze three common diagnostic methods for summer eczema (SE) in horses, an allergic dermatitis caused by bites of Culicoides spp. Nine horses with a medical history of SE and nine control animals were intradermally challenged with whole body extracts (WBE) and the saliva of a native (C. nubeculosus) and exotic (C. sonorensis) Culicoides species. Blood and serum samples of the horses were examined for basophil reactivity by a histamine release test (HRT) and for Culicoides-specific serum immunoglobulin E (IgE) and G (IgG) by enzyme-linked immunosorbent assay (ELISA). The results of intradermal testing (IDT) at 30min (immediate reactivity) and 4h (late-phase reactivity) post challenge with most insect preparations revealed significant differences between horses with and without SE. Overall, the HRT showed the most accurate results with a sensitivity of 1.00 for all Culicoides preparations and specificities of 0.78 (WBE) and 1.00 (saliva). By contrast, delayed reactions of the IDT (24h), and levels of Culicoides-specific IgE and IgG in the native serum showed little or no distinction between allergic and non-allergic horses. However, the use of purified serum IgE and IgG indicated the possibility for elevated titers of insect-specific serum immunoglobulins in horses with SE. The IDT and HRT did not reveal obvious differences in onset and intensity of positive reactions for the native verses exotic Culicoides species, whereas the ELISA showed slightly higher numbers of positive reactions for serum IgG with the indigenous species. Saliva, as compared to WBE, was found to have improved sensitivity and/or specificity for the HRT and for the late-phase immune reactions as measured by the IDT. Overall, the results indicate that allergy tests utilizing effector cells (mast cells, basophils) are more accurate in diagnosing SE in horses than serological analysis by ELISA. [Copyright &y& Elsevier]