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Background: The aim of the present study was to describe the presence of co-infection by Toxoplasma gondii and Neospora caninum in goats reared in extensive systems from Mexico. Materials and Methods: A cross-sectional study was conducted to determine the frequency of T. gondii and N. caninum, by detecting antibodies to each parasite by mean commercial ELISA kits. A total of 176 blood samples were randomly collected from mature females reared in extensive system herds from 20 municipalities of state of Guanajuato, Mexico. Results: The general seroprevalence was 23.9 and 21.0% for T. gondii and N. caninum, respectively, while co-infection rate was 3.6%. For geographic and environmental variables, no differences were observed among T. gondii and coinfection; however, it was observed that altitude, annual precipitation, annual average temperature, and rainy period showed significant differences with N. caninum seropositive goats. Conclusion: The seroprevalence of both parasites was appreciated in most of the studied herds. The present study is the first report of T. gondii and N. caninum co-infection in goats from extensive herds in Mexico. [ABSTRACT FROM AUTHOR]

Introduction: The transfer of immunoglobulins from the mother to newborns is widely recognized as a critical event for safeguarding offspring against potentially life-threatening infectious diseases. Mainly for this reason, this study aimed to assess the concentrations of immunoglobulin G (IgG) and immunoglobulin A (IgA) in the saliva of newborn calves and explore its potential use for monitoring passive immunity transfer from cows to calves, as also to evaluate how colostrum intake affects serum and saliva IgG and IgA concentrations. Methods: The quality of colostrum samples was evaluated using an optical refractometer before administration to the calves. Saliva and blood samples from 24 calves were obtained at the day of birth (T0) and 2 days after (T2) for determination of serum concentrations of total protein by refractometer, IgG and IgA (both on serum and saliva) by ELISA test. Results: Positive correlations were observed between salivary IgA at T2 and salivary IgG at T2. A significant increase in both IgG and IgA levels in calf serum and saliva was noted. Salivary IgA levels can reflect salivary IgG levels. Discussion: These findings suggest the potential utility of IgA in monitoring passive immunity transfer, and do not exclude saliva as an alternative, practical, and non-invasive matrix for assessing passive immunity transfer. [ABSTRACT FROM AUTHOR]

This study aimed to determine the sensitivity (Se) and specificity (Sp) of a circulating pathogen-specific biomarker (polyketide synthetase 5, Pks5)-based enzyme-linked immunosorbent assay (ELISA) independently or in conjunction with a caudal fold tuberculin (CFT) test for bovine tuberculosis (bTB) screening in dairy cattle. We enrolled 987 dairy cows from 34 herds in Chiang Mai province, Thailand. A conditionally independent Bayesian model with a single population was inferred from the test results. The percentage of positive results for the Pks5-ELISA using 0.4 OD cutoff test and CFT test were 9.0% (89/987) and 10.5% (104/987), respectively. The median of posterior estimates of Se for the Pks5-ELISA test was 90.2% (95% posterior probability interval [PPI] = 76.6-97.4%), while the estimated Sp was slightly higher (median = 92.9, 95% PPI = 91.0-94.5%). The median estimated Se of the CFT test was 85.9% (95% PPI = 72.4-94.6%), while the estimated Sp was higher, with a median of 90.7% (95% PPI = 88.7-92.5%). The posterior estimate for true disease prevalence was 2.4% (95% PPI = 1.2-3.9%). The Pks5-ELISA test yielded characteristics at or above the acceptable standards for bTB detection. Therefore, the pathogen-specific biomarker, Pks5, is a potential detection system for bTB screening and may be applied as an ancillary test together with the currently applied standard method (CFT test) to reinforce the bTB control and eradication programs. [ABSTRACT FROM AUTHOR]

Rapid and early detection of infectious diseases in pigs is important, especially for the implementation of control measures in suspected cases of African swine fever (ASF), as an effective and safe vaccine is not yet available in most of the affected countries. Additionally, analysis for swine influenza is of significance due to its high morbidity rate (up to 100%) despite a lower mortality rate compared to ASF. The wide distribution of swine influenza A virus (SwIAV) across various countries, the emergence of constantly new recombinant strains, and the danger of human infection underscore the need for rapid and accurate diagnosis. Several diagnostic approaches and commercial methods should be applied depending on the scenario, type of sample and the objective of the studies being implemented. At the early diagnosis of an outbreak, virus genome detection using a variety of PCR assays proves to be the most sensitive and specific technique. As the disease evolves, serology gains diagnostic value, as specific antibodies appear later in the course of the disease (after 7–10 days post-infection (DPI) for ASF and between 10–21 DPI for SwIAV). The ongoing development of commercial kits with enhanced sensitivity and specificity is evident. This review aims to analyse recent advances and current commercial kits utilised for the diagnosis of ASF and SwIAV. [ABSTRACT FROM AUTHOR]

Introduction: Different COVID-19 vaccines can affect various body systems, including the immune system, leading to distinctions in immune responses like natural killer cell cytotoxic activity, complement system function, and B-cell antibody production among vaccinated and nonvaccinated individuals. In Jordan, so far, there is no study exploring these immune response variations for four COVID-19 vaccines, along with the factors influencing the response and post-vaccination clinical outcomes. Methods: This study focuses on the humoral responses of 350 adult participants. Through participant interviews, one blood sample was collected for subsequent laboratory testing, utilizing the Enzyme-Linked Immunosorbent Assay (ELISA) method to examine the presence of COVID-19 antibodies in participants' plasma. A Chi-square test was performed to test for the statistical significance. Results: Our results indicate significant differences in positive antibody response efficiency (p value > 0.001) among participants who received two or three doses of COVID-19 inactivated and mRNA vaccines. The order of higher positive antibody frequency is as follows: Pfizer (90.5%), Sputnik V (88.1%), Sinopharm (78.8%), and AstraZeneca (65.8%). There is a noticeable inclination towards the efficacy of the Pfizer-BioNTech vaccine, evidenced by the highest levels of antibodies in recipients of this vaccine. Our study revealed that various factors, including gender, age, and smoking status prior to infection, play a role in significant contributors affecting immune response and influencing the effectiveness of these vaccines. Conclusion: All the four types of COVID-19 vaccines administered in Jordan demonstrated effectiveness. We recommend studying the ability of B lymphocytes present in the peripheral blood and the effect of Tregulatory cells by flow cytometry in recipients who have high positive titers. [ABSTRACT FROM AUTHOR]

According to the Domestic Animal Diversity Information System (DAD-IS) of the FAO, Italy has one of the largest numbers of local small ruminant breeds among European countries. In Southern Italy, namely the Campania Region, Bagnolese and Laticauda sheep breeds and Cilentana goat breeds are considered endangered according to the DAD-IS. Conservation of endangered animal breeds is a goal of the European Union (EU). However, the role of infectious diseases as risk factors for endangered breeds has rarely been considered. Small ruminant lentiviruses (SRLV) infect sheep and goats, causing slow-progressive, persistent, and debilitating diseases that can lead to animal death and productivity loss. In this study, we investigated the presence of SRLV in Bagnolese, Laticauda, and Cilentana breeds using a commercial ELISA in parallel with an in-house ELISA. The results of the two tests were in good agreement (Cohen Kappa 0.84, 95 % CI = 0.76–0.93). Discrepancies between the two tests were resolved using western blotting. In total, 430 samples were tested (248 Bagnolese, 125 Laticauda, and 57 Cilentana). The apparent prevalence rates were 12.5 %, 6.4 %, and 1.7 % in Bagnolese, Laticauda, and Cilentana, respectively. In the molecular analysis of 11 proviral partial sequences, subtypes B2 and A24 were identified in two Bagnolese herds. Owing to the beneficial role of sheep and goat breeding in marginal areas, it is important to screen the entire population and implement control/eradication of SRLV infections in conjunction with each conservation program.

This study aimed to determine the sensitivity (Se) and specificity (Sp) of a circulating pathogen-specific biomarker (polyketide synthetase 5, Pks5)-based enzyme-linked immunosorbent assay (ELISA) independently or in conjunction with a caudal fold tuberculin (CFT) test for bovine tuberculosis (bTB) screening in dairy cattle. We enrolled 987 dairy cows from 34 herds in Chiang Mai province, Thailand. A conditionally independent Bayesian model with a single population was inferred from the test results. The percentage of positive results for the Pks5-ELISA using 0.4 OD cutoff test and CFT test were 9.0% (89/987) and 10.5% (104/987), respectively. The median of posterior estimates of Se for the Pks5-ELISA test was 90.2% (95% posterior probability interval [PPI] = 76.6–97.4%), while the estimated Sp was slightly higher (median = 92.9, 95% PPI = 91.0–94.5%). The median estimated Se of the CFT test was 85.9% (95% PPI = 72.4–94.6%), while the estimated Sp was higher, with a median of 90.7% (95% PPI = 88.7–92.5%). The posterior estimate for true disease prevalence was 2.4% (95% PPI = 1.2–3.9%). The Pks5-ELISA test yielded characteristics at or above the acceptable standards for bTB detection. Therefore, the pathogen-specific biomarker, Pks5, is a potential detection system for bTB screening and may be applied as an ancillary test together with the currently applied standard method (CFT test) to reinforce the bTB control and eradication programs.

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Despite global efforts to assess the early response and persistence of SARS-CoV-2 antibodies in patients infected with or recovered from COVID-19, our understanding of the factors affecting its dynamics remains limited. This work aimed to evaluate the early and convalescent immunity of outpatients infected with SARS-CoV-2 and to determine the factors that affect the dynamics and persistence of the IgM and IgG antibody response. Seropositivity of volunteers from Mexico City and the State of Mexico, Mexico, was evaluated by ELISA using the recombinant receptor-binding domain (RBD) of the SARS-CoV-2 Spike protein for 90 days, at different time points (1, 15, 45, 60, and 90 days) after molecular diagnosis (RT-qPCR). Gender, age range, body mass index (BMI), comorbidities, and clinical spectrum of disease were analyzed to determine associations with the dynamics of anti-SARS-CoV-2 antibodies. On 90 days post-infection, individuals with moderate and asymptomatic disease presented the lowest levels of IgM, while for IgG, at the same time, the highest levels occurred with mild and moderate disease. The IgM and IgG levels were related to the clinical spectrum of disease, BMI, and the presence/absence of comorbidities through regression trees. The results suggest that the dynamics of anti-SARS-CoV-2 IgM and IgG antibodies in outpatients could be influenced by the clinical spectrum of the disease. In addition, the persistence of antibodies against SARS-CoV-2 could be related to the clinical spectrum of the disease, BMI, and the presence/absence of comorbidities. [ABSTRACT FROM AUTHOR]

The innovative product of 'Perilis Trading Ltd. 'BED Pro-biotic' is a probiotic neutralizer of mites and allergens in household dust. This product was tested for an inhibitory effect on the presence of mites of the species Dermatophagoides pteronyssinus in house dust. The data presented is a representative sample of 4 households and one office. The allergen DERPI is the major antigen produced by the house mite D. pteronyssinus, which is routinely found in house dust. The test for the presence of the allergen DERPI in the monitored premises was carried out before cleaning. After sampling, 'BED Pro-biotic' was administered and sampled again at 5-daily intervals before cleaning for one month. The product was applied according to the manufacturer's instructions. Antigen extraction was performed according to published protocols in the scientific literature, and antigen was detected with a specific immunoassay ELISA for the DERPI antigen. The result obtained showed that the product effectively reduces and eliminates the presence of D. pteronyssinus mites, which are common in Europe. [ABSTRACT FROM AUTHOR]

Rapid and early detection of infectious diseases in pigs is important, especially for the implementation of control measures in suspected cases of African swine fever (ASF), as an effective and safe vaccine is not yet available in most of the affected countries. Additionally, analysis for swine influenza is of significance due to its high morbidity rate (up to 100%) despite a lower mortality rate compared to ASF. The wide distribution of swine influenza A virus (SwIAV) across various countries, the emergence of constantly new recombinant strains, and the danger of human infection underscore the need for rapid and accurate diagnosis. Several diagnostic approaches and commercial methods should be applied depending on the scenario, type of sample and the objective of the studies being implemented. At the early diagnosis of an outbreak, virus genome detection using a variety of PCR assays proves to be the most sensitive and specific technique. As the disease evolves, serology gains diagnostic value, as specific antibodies appear later in the course of the disease (after 7–10 days post-infection (DPI) for ASF and between 10–21 DPI for SwIAV). The ongoing development of commercial kits with enhanced sensitivity and specificity is evident. This review aims to analyse recent advances and current commercial kits utilised for the diagnosis of ASF and SwIAV.

Despite global efforts to assess the early response and persistence of SARS-CoV-2 antibodies in patients infected with or recovered from COVID-19, our understanding of the factors affecting its dynamics remains limited. This work aimed to evaluate the early and convalescent immunity of outpatients infected with SARS-CoV-2 and to determine the factors that affect the dynamics and persistence of the IgM and IgG antibody response. Seropositivity of volunteers from Mexico City and the State of Mexico, Mexico, was evaluated by ELISA using the recombinant receptor-binding domain (RBD) of the SARS-CoV-2 Spike protein for 90 days, at different time points (1, 15, 45, 60, and 90 days) after molecular diagnosis (RT-qPCR). Gender, age range, body mass index (BMI), comorbidities, and clinical spectrum of disease were analyzed to determine associations with the dynamics of anti-SARS-CoV-2 antibodies. On 90 days post-infection, individuals with moderate and asymptomatic disease presented the lowest levels of IgM, while for IgG, at the same time, the highest levels occurred with mild and moderate disease. The IgM and IgG levels were related to the clinical spectrum of disease, BMI, and the presence/absence of comorbidities through regression trees. The results suggest that the dynamics of anti-SARS-CoV-2 IgM and IgG antibodies in outpatients could be influenced by the clinical spectrum of the disease. In addition, the persistence of antibodies against SARS-CoV-2 could be related to the clinical spectrum of the disease, BMI, and the presence/absence of comorbidities.

Corona viruses are a family of viruses that can cause diseases such as the common cold and acute respiratory infection and in 2019, a new type of corona virus was discovered that caused an outbreak of a disease that originated in China. The virus is known as severe acute respiratory syndrome corona virus (SARS-CoV-2). The resulting disease is called emerging corona virus disease 2019 (COVID-19). In March 2020, the World Health Organization declared the corona virus (COVID-19) to be a global pandemic. In this research, a diagnostic kit was prepared that is used in the laboratory to detect infection with Acquired Corona Virus (COVID19) by the method of the enzyme immunoassay (ELISA). Conjugated secondary antibodies tagged with HRP which gives a color signal with the substrate added to it, its intensity depends on the amount of antibodies present in the pathological sample. We used a microtiter plate coated with the virus core and ns antigen and a conjugate product from Imbian Company, while the other of the kit components (reagents and buffers) were prepared in Al-Razi laboratories to be suitable for use. Tests were conducted on the prepared kit for 96 samples, including 55 samples for positive cases and 41 samples for negative cases, which were obtained from the specialized laboratories and patients. The tests showed conformity in the results compared to foreign kit used for this purpose and using the ELISA washer and reader devices available in Al-Razi center's laboratories. And by installing the method of preparation by fixing the method of preparation and obtaining identical results, Al-Razi center can produce pioneering batches and provide the laboratories of Ministry of Health of this type of diagnostic kits.

This study was conducted for the purpose of evaluating infection with the parasite Fasciola spp in the city of Mosul for the period from June 2020 to June 2022. The results showed that a total of 12.30% of cattle and sheep had Fasciola eggs in their feces, with a significant difference between cattle and sheep, and the infection was high in cattle 16%. The eggs of the Fasciola species are oval, brown to yellow in color, and have an operculum. There was no significant difference in the rate of infection with the Fasciola parasite and the sex of cattle and sheep. Regarding the relationship between the rate of infection and the age of the animals infected, there were no significant differences between the infection and the age groups of the examined sheep. Whereas there was a significant difference between the cattle whose age was less than one year, one to two years old and more than two years old. The results of the serological examination of 90 blood samples of cattle and sheep showed that the seroprevalence of Fasciola in both cattle and sheep was 22.22% and 8.88%, respectively, with no difference in the incidence of infection between all the examined cattle and sheep. [ABSTRACT FROM AUTHOR]

The COVID-19 global pandemic is still affecting the world, even considering vaccine applications in most countries, especially due to new variant outbreaks and the possibility that they may present immunological escape. Therefore, mass testing is relevant in infection monitoring and restriction policy evaluations, making low-cost and easy-to-use tests essential. Serological tests might also be useful in monitoring immune response after vaccination. The present work proposes a less-expensive ELISA test route, using a scanner instead of a spectrophotometer and using the saturation of the image as a surrogate for the absorbance of each sample. Images from multiple experiments were selected and correlated with their spectrophotometric absorbance. ELISA plate images were digitized by a simple table scanner and, then, preprocessed using Hue, Saturation, Value (HSV) transformation, aiming to determine which correlates best with the obtained absorbance. Saturation correlated better with absorbance, and the experiments presented R 2 consistently above 90% between absorbance and the square of saturation. The new methodology showed similar accuracy, sensitivity, and specificity to the original method, all metrics ranging between 90% and 100% in most cases. An open-source software was also designed to analyze the images, perform the diagnosis, and generate reports. [ABSTRACT FROM AUTHOR]

Sarcoptic mange is a widely distributed disease, with numerous potential hosts among domestic and wild animals. Nowadays it is considered a neglected re-emergent infection in humans. As a difference with domestic pigs, and even with several clinical cases reported in some European countries, it seems that Eurasian wild boars (Sus scrofa) have a low susceptibility to clinical mange. However, because of a case of confirmed transmission from Spanish ibex (Capra pyrenaica) to wild boar in the province of Tarragona, we planned a large-scale ELISA survey in the neighboring Valencian Community (SE Spain). We compared 419 wild boar sera from different management systems (fenced vs. open game estates), different ages (piglets, juveniles, and adults), with different behaviour (gregarious females of all ages and male piglets vs. solitary juveniles and adult males), from areas with different wild boar densities, different wild ruminant densities and different sarcoptic mange epidemiologic situations. The whole prevalence of antibodies against sarcoptic mange in the tested wild boars was 10.5%. No significant differences were found when comparing fenced and free ranging wild boars, males and females, gregarious vs. solitary individuals or among different ages. However, wild boar density was a relevant factor. In areas with a hunting bag of <1 wild boar/km 2 , considered as a low density of suids, the seroprevalence was 2.94%, but rose to 11.52% in high density districts, constituting a significant difference (p = 0.037). Low wild boar populations would act as a protective factor (OR 0.233; p = 0.049) against coming into contact with the mite. The wild ruminant densities or their sarcoptic mange status did not show any effect on wild boars seroprevalence against this disease. These results reinforce the suggested host-taxon Sarcoptes scabiei specificity and the independence of host-species foci., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2024. Published by Elsevier Ltd.)

Aflatoxin contamination of maize in Rwanda and underlying causes are poorly documented. This study aimed to address this gap where maize samples for aflatoxins analysis were collected from two different agro-ecological zones and the temperature and relative humidity conditions during the time preceding sample collection were monitored to determine the conditions conducive to this contamination. Maize was analysed for moisture content and aflatoxin contamination and the temperature and humidity data were used to estimate the saturation deficit of the ambient air as an indicator of risk of contamination, which is high when the saturation deficit is low. Results showed an average lower saturation deficit of 0.0018 kg H₂O kg−1 in the high-altitude Volcanic region compared to the 0.00095 kg H₂O kg−1 recorded in the lower altitude region of the Eastern Savannah. Given the high moisture content in most of the analysed samples, this meant that it is more challenging to dry maize to the safe storage moisture content in the Volcanic region compared to the Eastern Savannah. The mean aflatoxin contamination was 40.45 μg kg−1 in the Volcanic region significantly higher than 4.66 μg kg−1 in the Eastern Savannah. Aflatoxin G1 was predominant among the contaminated samples. Affordable drying technologies that use renewable sources of energy should be developed and made accessible to farmer cooperatives. • Aflatoxins contamination in maize during storage is influenced by temperature and humidity conditions. • Low temperature drying of maize relies on temperature and humidity conditions in the environment. • Saturation deficit of air can be based on to assess the suitability of air to dry maize to the desired moisture content. • Low saturation deficit means prolonged drying time and thus increased risk of contamination. [ABSTRACT FROM AUTHOR]

Introduction Rhizomania viral disease is one of the limiting factors of sugar beet cultivation and significantly reduces its yield. The most important way to deal with this disease is to use resistant cultivars. Materials and Methods In order to evaluate the correlation between yield and qualitative traits of sugar beet cultivars with rhizomania disease indices, an experiment was conducted in a randomized complete block design with 12 different sugar beet cultivars in four replications in test site with natural infection with rhizomania in Torbat-e-Jam area at two years (2017 and 2018). The traits studied in this study included root yield, sugar content, sugar yield, white sugar content, white sugar yield, sodium, potassium, trace nitrogen, alkalinity, sugar extraction coefficient, leaf growth score, root growth score, disease severity and ELISA test index (OD). Analysis of variance was performed using SAS v.9.1 statistical software. Mean comparisons of traits were performed using LSD test at the probability level of 5 and 1%. Results and Discussion Analysis of variance showed significant differences among genotypes for all studied traits except for sodium. Cluster analysis of yield, crop quality as well as rhizomania severity of infection divided the sugar beet cultivars into three groups (resistant, semi-resistant and sensitive). Correlation between traits in a rhizomania infected field and different levels of resistance to rhizomania was different. In all cultivars with different levels of resistance, there was a significant correlation between pure sugar yield with leaf growth score (r=0.88 **), root growth score (r=0.85 **), disease severity (r= -0.84 **) and ELISA index (r= -0.67 **) was observed. Therefore, these traits are suitable indicators for evaluation of cultivars for suseptibility or resistance to rhizomania disease. In addition, correlation between ELISA index with disease severity (r=0.83**), leaf growth score (r= -0.61 **) and root growth score (r= -0.67 **) was significant. Conclusion According to the obtained results, because the ELISA test and determining the virus concentration is expensive, and on the other hand, this index had a positive and very significant correlation with the severity of the disease infection, so the disease severity index can be used to evaluate cultivars for resistance to rhizomania disease. [ABSTRACT FROM AUTHOR]

BACKGROUND: Histoplasmosis is caused by the dimorphic fungus Histoplasma capsulatum. In Mexico it is considered the systemic mycosis with the highest prevalence. CLINICAL CASE: A 24-year-old man began 3 months prior with a weight loss of 15 kg, unquantified intermittent fever, abdominal pain in the right upper quadrant and dyspnea. Based on the findings, an upper endoscopy and endoscopic retrograde cholangiopancreatography were performed, as well as a hemodialysis session due to anuria and severe metabolic acidosis. The patient's condition leads to respiratory failure, resulting in cardiorespiratory arrest and death. Postmortem histopathological analysis showed the presence of Histoplasma capsulatum in bone marrow, lung, liver, spleen, and kidney. ELISA postmortem serology for HIV was positive CONCLUSIONS: Disseminated histoplasmosis represents an entity with high morbidity and mortality. Histopathological diagnosis is essential to determine the final diagnosis when the other auxiliary diagnostic tests are inconclusive or negative and there is a high suspicion. [ABSTRACT FROM AUTHOR]

Background & Objective: Thiopurine drugs are considered as a treatment modality in various autoimmune disorders including pemphigus vulgaris (PV). These drugs are metabolized by an enzyme “Thiopurine S-methyl transferase” (TPMT). Various variants of this enzyme may have decreased activity leading to serious drug side effects. To investigate the phenotype and genotype of TPMT in PV patients receiving thiopurine drugs. Methods: 50 patients (29 women and 21 men) with pemphigus vulgaris treating with standard dose of Thiopurine drugs were selected. Sex, age, result of liver function test and complete blood count were recorded. Genotyping of two common non-functional allele (TPMT*2 and TPMT*3C) by Allele-specific and RFLP-PCR was performed. TPMT enzymatic level was determined by an ELISA based method. Results: Of patients, 36 (72%) were found to have normal TPMT level; and 12, (24%) had higher level of enzyme and 2, 4% had low TPMT enzyme, but none of the patients showed mutant TPMT*2 and TPMT*3C alleles. None of the patients showed hepatotoxicity and bone marrow suppression. Conclusion: The phenotypic assay based on ELISA method may have false positive and misleading results but genotyping using PCR-RFLP and allele specific PCR is accurate, simple and cost-effective and can be used in patients decided to undergo thiopurine treatment.

Lysozyme is a biologically active protein present in egg whites, having an essential role in the immune defense of the embryo. Due to its antimicrobial properties, it is extracted and used in the composition of various products as a preservative and/or a bioactive ingredient. Its qualitative and quantitative analysis can be performed by various test methods widely presented in the literature. The present paper presents the results obtained in assessing the lyzozyme content of aqueous solutions derived from hyperimmune egg white and/or from biologically active products in which it was included by using the sandwich ELISA technique.

Abstract Background Brucellosis is a zoonosis whose incidence is not declining worldwide despite the global effort to control the disease. Accurate and precise diagnosis is a crucial step in any prophylaxis program but single tests to unequivocally detect animals infected with Brucella spp. are currently unavailable. In Italy, serological diagnosis of bovine brucellosis is performed with two official tests: a rapid agglutination test (i.e., Rose Bengal Plate test, RBPT) and a complement fixation test (CFT) that detect antibodies directed mainly to the smooth lipopolysaccharide (S-LPS). Neither of the two tests is able to avoid the detection of false positive serological reactions (FPSRs) caused by bacteria sharing S-LPS components with Brucella spp. and responsible for the single reactors (SR) phenomenon. A B. melitensis R strain-based ELISA showed a good diagnostic performance in unravelling FP animals; however, since a limited number of animals were analyzed in that study, a large field study was conducted here to discriminate between Brucella-infected from FP animals, with the final aim of reducing the unnecessary slaughter of the latter. An ELISA based on a R strain of Brucella, i.e., Brucella melitensis B115, was employed to measure specific IgG responses in a collection of bovine sera (n = 648). Sera were obtained from 180 farms (either officially brucellosis-free or not brucellosis-free) recruited during an extended period of time (2014–2018) and were preliminarily assayed with the official tests by the Italian Reference Centers and then subjected to the ELISA. Results Negative sera, when subjected to the ELISA, gave O.D. values below the cutoff; SR sera, i.e. RBPT positive and CFT negative, as well as double positive (DP) sera, i.e. RBPT and CFT positive, gave O.D. values that were below the cutoff. All positive sera, i.e. from Brucella-infected animals, were RBPT positive and CFT positive (ICFTU ranging from 20 to 1280) and gave ELISA O.D. values above the cutoff. Conclusions The B. melitensis B115-based ELISA systematically unravelled all false positive (FP) sera while confirming the diagnosis in Brucella-infected animals. Thus, the test employed in the present study may complement the official assays to avoid the costly slaughter of FP animals.

West Nile fever is a vector borne viral disease that can affect humans, horses, birds and sometimes other species of animals. Every year West Nile fever is detected in the human population in Serbia. The disease often occurs in a subclinical form, but most clinically evident cases occur in horses. Therefore, horses are recommended as a sentinel species for monitoring the general incidence of West Nile fever in a specific territory. Our goal was to determine the prevalence of antibodies against West Nile fever virus in horses in the Belgrade epizootiological area. We examined serum samples from 77 horses to determine the seroprevalence of West Nile fever virus in horses throughout the city of Belgrade. Sera were tested by commercial ELISA tests for detection of specific IgG-class antibodies to West Nile fever virus and for the detection of specific IgM-class antibodies to confirm the presence of old and acute (recent) infections in horses. The results confirmed that West Nile fever virus is widespread, detected in 70.1% of the surveyed horse population in Belgrade, and we also detected 5.1% of acute cases had occurred due to horses being infected in 2019. The seroprevalence of West Nile virus in the horse population in the municipality of Belgrade is increasing.

According to the Domestic Animal Diversity Information System (DAD-IS) of the FAO, Italy has one of the largest numbers of local small ruminant breeds among European countries. In Southern Italy, namely the Campania Region, Bagnolese and Laticauda sheep breeds and Cilentana goat breeds are considered endangered according to the DAD-IS. Conservation of endangered animal breeds is a goal of the European Union (EU). However, the role of infectious diseases as risk factors for endangered breeds has rarely been considered. Small ruminant lentiviruses (SRLV) infect sheep and goats, causing slow-progressive, persistent, and debilitating diseases that can lead to animal death and productivity loss. In this study, we investigated the presence of SRLV in Bagnolese, Laticauda, and Cilentana breeds using a commercial ELISA in parallel with an in-house ELISA. The results of the two tests were in good agreement (Cohen Kappa 0.84, 95 % CI = 0.76-0.93). Discrepancies between the two tests were resolved using western blotting. In total, 430 samples were tested (248 Bagnolese, 125 Laticauda, and 57 Cilentana). The apparent prevalence rates were 12.5 %, 6.4 %, and 1.7 % in Bagnolese, Laticauda, and Cilentana, respectively. In the molecular analysis of 11 proviral partial sequences, subtypes B2 and A24 were identified in two Bagnolese herds. Owing to the beneficial role of sheep and goat breeding in marginal areas, it is important to screen the entire population and implement control/eradication of SRLV infections in conjunction with each conservation program., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Abstract Background Streptococcus pneumoniae infection causes morbidity and mortality in multiple myeloma patients. Pneumococcal vaccination is commonly given to immunocompromised myeloma patients; however response data are sparse. Here, we present longitudinal response data to pneumococcal vaccination in multiple myeloma patients. Method Twenty‐eight multiple myeloma patients were included, 25 of whom were newly diagnosed. All the patients received two vaccines Prevnar13® and Pneumo23®. Serotype‐specific IgG was measured by ELISA for all 23 vaccine serotypes at baseline, and then sequentially at different time points postvaccination until treatment ended. Response to vaccination is available for 20 patients. The primary endpoint was the incidence rate of patients who obtained an isotype response serum concentration after vaccination. Secondary endpoints included detailed isotype increase, time to first increase, further assessment of a decreased anti‐pneumococcal serum concentrations following treatment including autologous stem cell transplantation (ASCT), rate of infection with a special attention to pneumococcal infection. Results The median age was 66 years and the male to female ratio was 0.6. Anti‐pneumococcal capsular polysaccharide (anti‐PCP23) IgG, IgG2, IgA, and IgM responses were detected within 1 week postvaccination. Response to at least one subtype of antibody was obtained in 85% (n = 17) of patients, for at least two subtypes in 65% (n = 13), for at least three subtypes in 55% (n = 11), and 2 patients responded to all four subtypes. The median increase in the concentration of anti‐PCP23 isotypes was threefold following vaccination, with the highest increase observed when Pneumo23® was given more than 30 days after Prevnar13®. The anti‐pneumococcal geometric mean concentration decreased significantly for all subtypes over time independently of treatment approaches. Conclusion Myeloma has the ability to demonstrate a response to pneumococcal vaccine, independently of preexisting hypogammaglobulinemia and possibly of treatment‐induced immunodepression. We also observed a drop in the serum response overtime and following autologous transplantation. Further studies in larger sample are needed to understand the benefit of vaccination strategies in these patients.

Abstract Background The objective of this study was to determine the sensitivity (Se) and specificity (Sp) of bovine tuberculosis (bTB) screening tests including a single intradermal tuberculin (SIT) test, interferon gamma (IFN-γ) assay, and a commercial ELISA test (M. bovis Ab) in dairy cattle, under field conditions, using a Bayesian approach. Results The study population consisted of 128 dairy cows from 25 bTB-infected herds in Chiang Mai and Chiang Rai provinces, Thailand. A single-population Bayesian model was implemented assuming conditional dependence between the SIT test and IFN-γ assays. The 95% posterior probability interval (PPI) of the SIT test (severe interpretation) Se ranged from 75.3 to 95.2% (median = 87.6%), while the Sp was slightly lower (median = 83.6%, PPI = 74.2–92.8%). The IFN-γ assay Se was moderate and the 95% PPI ranged from 38.6 to 74.4% (median = 55.7%) with higher Sp (median = 93.5.4%, PPI = 87.0–98.1%). The M. bovis Ab ELISA Se was low, with 95% PPI ranging between 30.0 and 71.2% (median = 47.4%); however, the Sp was high (median = 90.9%, PPI = 84.5–95.5%). Conclusion The SIT test sensitivity was similar to that demonstrated in other regions and can, therefore, be used effectively as part of control programs in this area. The IFN-γ and M. bovis Ab ELISA assays can be applied as supplementary techniques. The test performance of these tests when used as single tests without confirmation, however, are expected to continue to challenge disease eradication efforts.

In recent years, numerous reports have repeatedly highlighted a series of food safety scandals involving contaminated and forged meat and fish products, grains and fruit products, juices, cooking oils, and spices and herbs, distilled beverages and pet treats. Foods that have been found to be stained with chemicals, illicit drug residues, additives and dyes, pathogenic microorganisms and other pests. Some foods enter the market have expired well or are unhygienic. Economic gain is the goal of food fraud. Food fraud and the prevention of such fraud are very important processes in the food industry. Such frauds are economically motivated, rated as criminal behaviour, and the moment we understand criminal behaviour and decision making we will be able to calculate and exclude the risk of food fraud. By analysing financially motivated fraud that combine opportunities, motivations, and inadequate control measures, we can assess the likelihood of fraud in any food product or component. The modified ingredients are specially designed to avoid quality assurance and quality control systems for customers. Only people who manipulate the ingredients know what substances and how to manipulate them. In addition, fraudulent ingredients are often unconventional substances that do not meet the requirements of food safety management systems, and become known only after they are incorporated into the supply chain. International standards for food and regulations address the risk of fraud food adulteration. European Union (EU) Directives, Global Food Safety Initiative (GFSI) Codex Alimentarius has continued work on a food fraud, or how, food fraud fits into their benchmarking. The problem of detection and typing of meat in meat products in the world and lack of research on them in the Republic of North Macedonia was the main goal for this paper. Our task was identification of meat type by ELISA method and proof of counterfeiting of meat products. Analyses are made in the laboratories of the Institute of Food at the Faculty of veterinary medicine in Skopje A total of 350 samples of various heat-treated meat products subgroups of meat sausages in pieces were examined for detection, typifying and quantifying the type of meat used for production.

Objective: To identify the serological epidemiology of Cryptosporidium infections and to follow up on the changes in the infection profile in Southern Egypt in order to establish a suitable scheme for control and prevention of cryptosporidiosis. Methods: A total of 1 912 (960 from human and 952 from animals) stool specimens and sera were screened for Cryptosporidium species using modified Ziehl Neelsen technique and a newly-developed enzyme-linked immunosorbent assay (ELISA). Environmental risk factors and socioeconomic data were surveyed by questionnaire between September 2016 and December 2017. Results: Totally, 20.83% of the human subjects were positive for Cryptosporidium infection tested by ELISA. The seropositivity was positively correlated with age. The prevalence of Cryptosporidium infections in females was significantly higher than in males (P

Aim of study: To report the occurrence of Pepino mosaic virus (PepMV) on tomato in Serbia and to genetically characterize Serbian PepMV isolates. Area of study: Tomato samples showing virus-like symptoms were collected in the Bogojevce locality (Jablanica District, Serbia). Material and methods: Collected tomato samples were assayed by DAS-ELISA using antisera against eight economically important or quarantine tomato viruses. Three selected isolates of naturally infected tomato plants were mechanically transmitted to tomato ‘Novosadski jabučar’ seedlings. For confirmation of PepMV infection, RT-PCR was performed using specific primers PepMV TGB F/PepMV UTR R. Maximum-likelihood phylogenetic tree was constructed with 47 complete CP gene sequences of PepMV to determine the genetic relationship of Serbian PepMV isolates with those from other parts of the world. Main results: The results of DAS-ELISA indicated the presence of PepMV in all tested samples. Mechanically inoculated ‘Novosadski jabučar’ seedlings expressed yellow spots and light and dark green patches, bubbling, and curled leaves. All tested tomato plants were RT-PCR positive for the presence of PepMV. The CP sequence analysis revealed that the Serbian PepMV isolates were completely identical among themselves and shared the highest nucleotide identity of 95.1% (99.2% aa identity) with isolate from Spain (FJ263341). Phylogenetic analysis showed clustering of the Serbian PepMV isolates into CH2 strain, but they formed separate subgroup within CH2 strain. Research highlights: This is the first data of the presence of PepMV in protected tomato production in Serbia. Considering increased incidence and rapid spread in Europe, the presence of PepMV on tomato could therefore represent serious threat to this valuable crop in Serbia.

Background: The crevicular fluid analysis represents a useful diagnosis tool, with the help of which noninvasive cellular metabolic activity can be analyzed. The aim of the study is to investigate comparatively IL1β and IL6 in the gingival crevicular fluid of clinically healthy adolescents and young adults during the acute phase of orthodontic treatment. Methods: Gingival crevicular fluid was collected from 20 patients (aged between 11 and 28) undergoing orthodontic treatment. Measurements were taken before (T0) and after 24 h after distalization forces were activated (T1). IL1β and IL 6 were analyzed using Elisa tests. The statistical tests used were two-sided t tests. Results: Between the two time periods there was a significant raise both in the crevicular fluid rate (0.57 µL at T0 vs. 0.95 µL at T1, p = 0.001) and in IL1β levels (15.67 pg/µL at T0 vs. 27.94 pg/µL at T1, p = 0.009). We were able to identify IL6 only in a third of the sites. There is a significantly increased level of ILβ at T1 in adolescents, more than in young adults (42.96 pg/µL vs. 17.93 pg/µL, p = 0.006). Conclusions: In the early stage of orthodontic treatment, the periodontal tissues of adolescents are more responsive to orthodontic forces than those of young adults. [ABSTRACT FROM AUTHOR]

Few data are available about the prevalence of Salmonella enterica serovar Enteritidis (S.E.) and Salmonella enterica serovar Typhimurium (S.T.) in ornamental poultry in Italy. The aim of this study was to investigate the seroprevalence for S.E. and S.T. using serological tests and the prevalence of Salmonella spp. in eggs by culture methods. For this purpose, 240 serum samples and 216 eggs were sampled from asymptomatic and unvaccinated ornamental hens reared in 24 farms, located in 8 different Italian regions. As screening test, a Tube Serum Agglutination test (TSA) was performed on 231 out of 240 serum samples. Four out of 24 farms (16.67%) were serologically positive for Salmonella spp. for a total of 10 samples. These positive samples were confirmed using an ELISA test and the results show that 5/231 (2.16%) and 7/231 (3.03%) serum samples were positive for S.E. and S.T. respectively, and 2/231 (0.87%) for both serotypes. Among all farms, 2/24 (8.33%) were positive for S.E. and 4/24 (16.67%) for S.T. The analysis of eggs using culture methods gave negative results for both yolk and shell pools (0/48, 0.0%). The seroconversion associated with exposure to S.E./S.T. in ornamental poultry, poses a potential public health problem. This study confirms that S.E. and S.T. are widespread in studied backyard poultry farms as asymptomatic form, and animals as potential reservoirs of Salmonella. It is necessary to inform farmers that a regular and periodic control of animals, eggs or meat, is very important to prevention of Salmonella foodborne infections and their spread. [ABSTRACT FROM AUTHOR]

An indirect variant of ELISA used for detection of antibodies to nonstructural proteins of the FMD virus in porcine blood sera was developed. The results of the validation showed that the developed method is characterized by high sensitivity, specificity and reproducibility. When testing the blood serum panel obtained from experimentally infected animals, the method allowed to detect antibodies to FMD virus in 7 of 18 sera collected on day 6 post inoculation, in 13 of 19 sera – on day 7 post inoculation, in 16 of 19 sera – on day 8 post inoculation and in all 76 sera obtained on days 9–12 post inoculation. The diagnostic specificity of 3AB-ELISA was 100% when testing 100 knowingly negative blood sera from pigs imported to Russia from Norway. High specificity and sensitivity of the method, established during the development of the method, are confirmed in the course of routine diagnostic tests.

Introduction Syphilis (lues) is a chronic infectious disease caused by the spirochete bacteria Treponema pallidum. The infection is most commonly transmitted by sexual contact, more rarely via the transplacentar route from the infected mother to the baby, while the transmittion via blood and/or blood products or accidental innoculation is at this day and age extremely rare. During the period of time examined in this report, the number of incidents of syphilis infection in Serbia has tripled. Diagnostic algorithms for syphilis are nowadays prone to modifications. World Health Organization (WHO) and Center for Disease Control (CDC) guidelines are modified in many European countries to better suit local conditions. Objective: Presentation and analysis of ELISA serology tests for syphilis performed at the Clinic for Infectious and Tropical Disease, Clinical Center of Serbia, for the period of 2004 to 2016 and comparisson of these results with data obtained from the Institute of Public Health of Serbia 'Dr Milan Jovanović Batut' for the same time period. Methodology: During the afore mentioned period of time, 252.310 patients were tested for syphilis, using the 3rd generation ELISA test. Those results were compared with data obtained from the Institute of Public Health of Serbia for the same time period. Results: A trend of growth has been noted for new infections and incidence of disease for syphilis almost 4.5 times between 2004 (35 new infections, incidence of disease 0.5/100.000 people) and 2016 (155 new infections, incidence of disease 2-21/100.000 people). The results of our study are compatible with the results published in the yearly report of the Institute of Public Health of Serbia for 2016, stating that the number of people diagnosed with syphilis (159) in 2016 is 9.23% higher than in 2015 (146 patients), which makes the registered five-year period of 2012-2016 the time period with the highest registered rate of incidence of syphilis infection. Conclusion: Syphilis (lues) is one of the most serious chronic sexually transmittive diseases, caused by the spirochete bacteria Treponema pallidum. A trifold rise in the incidence of syphillis infection has been noted in the previous years as shown by our study performed at the Clinic for Infectious and Tropical Disease, Clinical Center of Serbia. Diagnostics for syphilis should be performed according to the precise algorithm prescribed by the World Health Organization, adapted to the conditions of testing in Serbia.

Infections caused by viruses hepatitis B and C are one of the most severe problem in contemporary medicine. These viruses show high affinity to hepatocytes. The fundamental pathogenetic mechanisam which is responsible for hepatocyte damage, depends either on cytocidal effect of virus or immunological response of the infected host. The most common way of virus transmission nowdays is parenteralno, due to intravenous drug abuse, while earlier the blood transfusion was a more often the pathway of infection. Patients on chronical hemodialysis are at increased risk of HCV infection. The aim of our study was to determine the most common risk factors and pathways of HBV and HCV infection, as well as to assess the significance of the serological markers of hepatitis B and hepatitis C viruses in risk groups of patients. The study was conducted at Institute for blood transfusion at Health Center of Kosovska Mitrovica, in the period from January 2000 until December 2015. ELISA test was performed for the confirmation of presence of hepatitis B and hepatitis C viruses in the blood. The results showed that the number of respondents increased significantly during the research period. Voluntary blood donors were significantly more represented than respondents who were referred by a physician. Average age of all seropositive? examinees in the study was 47,1 ± 18,1 years (range from 8 - 79 years). HBV seropositivity was significantly more present compared to HCV seropositivity. The highest incidence of HBV and HCV infection was recorded in the working age population, between 40 and 50 years of age. Analyzing the data of the way of infection transmission, most patients reported an unknown pathway of infection (39%). The second most common way of infection transmission was hemodialysis (28.4%), while the intravenous drug abuse was noted in 13,4% of patients. We can conclude that the implementation of regular screening and timely prevention, as well as the education program of the population can be helpful in reducing the number of patients with HBV and HCV infection.

Background & Objective: Thiopurine drugs are considered as a treatment modality in various autoimmune disorders including pemphigus vulgaris (PV). These drugs are metabolized by an enzyme “Thiopurine S-methyl transferase” (TPMT). Various variants of this enzyme may have decreased activity leading to serious drug side effects. To investigate the phenotype and genotype of TPMT in PV patients receiving thiopurine drugs. Methods: A total of 50 patients (29 women and 21 men) with pemphigus vulgaris treating with standard dose of Thiopurine drugs were selected. Sex, age, result of liver function test and complete blood count were recorded. Genotyping of two common non-functional allele (TPMT*2 and TPMT*3C) by Allele-specific and RFLP-PCR was performed. TPMT enzymatic level was determined by an ELISA based method. Results: Of patients, 36 (72%) were found to have normal TPMT level; and 12, (24%) had higher level of enzyme and 2, 4% had low TPMT enzyme, but none of the patients showed mutant TPMT*2 and TPMT*3C alleles. None of the patients showed hepatotoxicity and bone marrow suppression. Conclusion: The phenotypic assay based on ELISA method may have false positive and misleading results but genotyping using PCR-RFLP and allele specific PCR is accurate, simple and cost-effective and can be used in patients decided to undergo thiopurine treatment. [ABSTRACT FROM AUTHOR]

Background: Accurate HIV diagnosis is essential for appropriate patient care. Rapid tests (RTs) are considered key to HIV screening and management. Some studies have found RTs to be comparable with the ELISA test whilst others have reported lower sensitivity. Aim and study design: The aim of this retrospective, descriptive study was to evaluate the sensitivity and specificity of the HIV 1/2/O Tri‐line rapid test (HIV‐TRT) device compared with ELISA. Method: The study sample comprised 45 records of patients who tested for HIV using the HIV‐TRT device and ELISA. Results: As compared with ELISA as the 100% gold standard, the sensitivity of the HIV‐TRT was 80% (CI: 59%–93%) and specificity was 100% (CI: 83%–100%). ROC area of 0.9 at 95% CI was determined. Conclusion: The low sensitivity of HIV‐TRT is a concern, since HIV screening in South Africa makes use of RTs. [ABSTRACT FROM AUTHOR]

Parietaria mottle virus (PMoV) is considered an emerging virus in many countries of the Mediterranean basin. Outbreaks of tomato necrotic disease caused by the tomato strain PMoV‐T have been recorded in Italy, France, Spain, and Greece. No resistant or tolerant tomato cultivars are available and no cases of resistance have been reported in the section Lycopersicon of the Solanum genus so far. Therefore, wild relatives of tomato were evaluated to identify potential sources of resistance to PMoV infection. In all, 16 germplasm accessions from six wild Solanum spp. of the section Lycopersicon, along with susceptible near‐isogenic tomato lines (Monalbo, Momor, and Mospomorist), were evaluated for resistance against two different PMoV‐T strains, TI1 (from Italy) and FR1 (from France), under greenhouse conditions using mechanical inoculations. Local and systemic virus infections were determined by enzyme‐linked immunosorbent assay (ELISA) 15 and 30 days after mechanical inoculation and confirmed by dot‐blot hybridization of viral RNA with a PMoV‐specific digoxigenin‐labelled riboprobe. No local or systemic symptoms were observed in three accessions of S. peruvianum and one accession of S. corneliomuelleri. Moreover, the virus was never detected in the inoculated and upper noninoculated leaves of these accessions, either with ELISA or the specific riboprobe. The accession of S. lycopersicum var. cerasiformae was resistant only to the TI1 strain. This is the first description of sources of extreme resistance to PMoV in the section Lycopersicon of the Solanum genus. [ABSTRACT FROM AUTHOR]

This stiidy was conducted to evaluate prevalence of BVD and the strategies which were applied for prevention of BVD over the past 50 years in some provinces of Iran. Blood samples of 500 Iranian Holstein race were randomly collected from three dairy farms located in Tehran, Isfahan and Qazvin provinces. BVD conteol protocols of these farms were recorded. ELISA technique was used to measure the antibody and antigen titers for BVD. RT-PCR technique was performed to investigate the presence and the type of virus in all samples. The prevalence of Ab-/ Ag-, Ab+/ Ag-, Ab-/ Agt and Ab-/ Agt were 10.2%, 78.8%, 7.2% and 3.8%, respectively. Furthermore, approximately 4.2% of the animals were PI and the prevalence of antibody and antigen titers had not significant difference in three provinces. All positive samples were BVDV type 1. Type 2 was not observed in this stiidy. The results of the stiidy indicated tiiat the efficiency of the used protocols to conteol BVD diseases is not successfill. So, to conteol the BVD, applying large management policies at the national level is fundamental beside the bioseeurity stiategies. [ABSTRACT FROM AUTHOR]

Lysozyme is a biologically active protein present in egg whites, having an essential role in the immune defense of the embryo. Due to its antimicrobial properties, it is extracted and used in the composition of various products as a preservative and/or a bioactive ingredient. Its qualitative and quantitative analysis can be performed by various test methods widely presented in the literature. The present paper presents the results obtained in assessing the lyzozyme content of aqueous solutions derived from hyperimmune egg white and/or from biologically active products in which it was included by using the sandwich ELISA technique. [ABSTRACT FROM AUTHOR]

One of the main objectives of root canal treatment is to alleviate the pain associated with irreversibly inflamed pulps. However, some patients may have moderate to severe pain following treatment. The aim of this study was to compare and assess the effect of ketorolac tromethamine on substance P expression in the pulp and periapical tissues when used as a root canal irrigant for single‐visit root canal treatment in teeth with irreversible pulpitis. Thirty‐six patients were randomly allotted to three irrigant groups – saline (n = 14), 3% sodium hypochlorite (n = 11) and ketorolac tromethamine (n = 11). Pulp blood samples (S1) were collected on gaining access to the pulp, and periapical blood samples (S2) were collected after root canal preparation. Quantification of substance P was done by ELISA test. The ketorolac tromethamine group had greater reduction in substance P expression (S2). Post‐operative pain levels were not significantly influenced by the different root canal irrigants. [ABSTRACT FROM AUTHOR]

Background: Brucellosis is a zoonosis whose incidence is not declining worldwide despite the global effort to control the disease. Accurate and precise diagnosis is a crucial step in any prophylaxis program but single tests to unequivocally detect animals infected with Brucella spp. are currently unavailable. In Italy, serological diagnosis of bovine brucellosis is performed with two official tests: a rapid agglutination test (i.e., Rose Bengal Plate test, RBPT) and a complement fixation test (CFT) that detect antibodies directed mainly to the smooth lipopolysaccharide (S-LPS). Neither of the two tests is able to avoid the detection of false positive serological reactions (FPSRs) caused by bacteria sharing S-LPS components with Brucella spp. and responsible for the single reactors (SR) phenomenon. A B. melitensis R strain-based ELISA showed a good diagnostic performance in unravelling FP animals; however, since a limited number of animals were analyzed in that study, a large field study was conducted here to discriminate between Brucella-infected from FP animals, with the final aim of reducing the unnecessary slaughter of the latter. An ELISA based on a R strain of Brucella, i.e., Brucella melitensis B115, was employed to measure specific IgG responses in a collection of bovine sera (n = 648). Sera were obtained from 180 farms (either officially brucellosis-free or not brucellosis-free) recruited during an extended period of time (2014–2018) and were preliminarily assayed with the official tests by the Italian Reference Centers and then subjected to the ELISA. Results: Negative sera, when subjected to the ELISA, gave O.D. values below the cutoff; SR sera, i.e. RBPT positive and CFT negative, as well as double positive (DP) sera, i.e. RBPT and CFT positive, gave O.D. values that were below the cutoff. All positive sera, i.e. from Brucella-infected animals, were RBPT positive and CFT positive (ICFTU ranging from 20 to 1280) and gave ELISA O.D. values above the cutoff. Conclusions: The B. melitensis B115-based ELISA systematically unravelled all false positive (FP) sera while confirming the diagnosis in Brucella-infected animals. Thus, the test employed in the present study may complement the official assays to avoid the costly slaughter of FP animals. [ABSTRACT FROM AUTHOR]

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Michael J Cook,1 Basant K Puri2 1Independent Researcher, Highcliffe, 2Department of Medicine, Hammersmith Hospital, Imperial College London, London, UK Abstract: In this study, Bayes’ theorem was used to determine the probability of a patient having Lyme disease (LD), given a positive test result obtained using commercial test kits in clinically diagnosed patients. In addition, an algorithm was developed to extend the theorem to the two-tier test methodology. Using a disease prevalence of 5%–75% in samples sent for testing by clinicians, evaluated with a C6 peptide enzyme-linked immunosorbent assay (ELISA), the probability of infection given a positive test ranged from 26.4% when the disease was present in 5% of referrals to 95.3% when disease was present in 75%. When applied in the case of a C6 ELISA followed by a Western blot, the algorithm developed for the two-tier test demonstrated an improvement with the probability of disease given a positive test ranging between 67.2% and 96.6%. Using an algorithm to determine false-positive results, the C6 ELISA generated 73.6% false positives with 5% prevalence and 4.7% false positives with 75% prevalence. Corresponding data for a group of test kits used to diagnose HIV generated false-positive rates from 5.4% down to 0.1% indicating that the LD tests produce up to 46 times more false positives. False-negative test results can also influence patient treatment and outcomes. The probability of a false-negative test for LD with a single test for early-stage disease was high at 66.8%, increasing to 74.9% for two-tier testing. With the least sensitive HIV test used in the two-stage test, the false-negative rate was 1.3%, indicating that the LD test generates ~60 times as many false-negative results. For late-stage LD, the two-tier test generated 16.7% false negatives compared with 0.095% false negatives generated by a two-step HIV test, which is over a 170-fold difference. Using clinically representative LD test sensitivities, the two-tier test generated over 500 times more false-negative results than two-stage HIV testing. Keywords: false-positive test, false-negative tests, probability of disease, serology testing methodology, Lyme borreliosis, Bayes’ theorem, two-tier test, test sensitivity, ELISA test, Western blot test, HIV testing

The current study was conducted in Thi-Qar province to investigate of Toxoplasma gondii prevalence among renal dialysis and cancer patients by using anti-Toxoplasmosis IgG and IgM by ELISA also the study determined of some factors that may affected the prevalence of infection, such as age, sex, marital status, place of residence and a number of chemical dosing that exposed to it cancer patient or the number of times the dialysis for renal dialysis patients. The results of the current study showed that the prevalence of toxoplasmosis in Thi-Qar province in both renal dialysis and cancer patients and control group were 28% and 36% and 24% respectively. recorded results of Enzyme Linked Immunosorbent Assay ELISA test results for a positive antibody IgG in both renal dialysis and cancer patients and control group 25% , 30% , 22% respectively. As for the IgM antibody study did not record any positive result in both categories of the study (dialysis and cancer) only control group was 2%. The result of IgG & IgM together in renal dialysis and cancer and control group 3%, 6%, 0% respectively. The study found that the increase in the number of times the dialysis increased incidence of infection prevalence it was 25.53% for patients who conducted the dialysis treatment more than six times and 16.66% for patients who have less than six times.For factor chemical dosing show high infection rates in patients who have been exposed to more than one dose of a chemical 37.63% As for those who were not exposed to any chemical dose was ratios have 14.29% .