ID Screen Research Articles

Detection of antibodies to Toxoplasma gondii and Neospora caninum in animals from three zoos in Slovakia.

The aim of this study was to detect antibodies to Toxoplasma gondii and Neospora caninum in exotic animal species kept in three zoos in Slovakia. Antibodies to T. gondii and N. caninum were detected by commercial Enzyme-linked immunosorbent assay (ELISA, ID Screen Toxoplasmosis Indirect Multispecies and ID Screen Neospora caninum Indirect Multispecies, ID Vet, Montpellier, France). Antibodies to T. gondii and N. caninum were detected in 43% (24/56) and 5% (3/55) of animals, respectively. The three animals with N. caninum antibodies: two wolves (Canis lupus) and one Hartmann's mountain Zebra (Equus zebra hartmannae), were clinically healthy, and both wolves simultaneously had antibodies to T. gondii. The results of our study provide a picture of the recent circulation of T. gondii in three Slovakian zoos with the S/P (ratio of antibodies in the sample to antibodies in positive control) value higher than 200%, found in five animals (9%) indicating acute toxoplasmosis. The highest S/P value (296%) was detected in a Roloway monkey (Cercopithecus roloway), which was healthy without clinical signs, presuming that Roloway monkey is a species less susceptible to T. gondii infection. Results of our study showed the presence of T. gondii and N. caninum in Slovakian zoos, confirming recent T. gondii infections according to the high level of antibodies detected in five animals, referring to acute toxoplasmosis.

Comparison of Serological Methods for Tick-Borne Encephalitis Virus-Specific Antibody Detection in Wild Boar and Sheep: Impact of the Screening Approach on the Estimated Seroprevalence.

Tick-borne encephalitis virus (TBEV) is a flavivirus transmitted by ticks. Serological screenings in animals are performed to estimate the prevalence and distribution of TBEV. Most screenings consist of a primary screening by ELISA, followed by confirmation of positive samples by plaque reduction neutralization tests (PRNTs). In this study, 406 wild boar sera were tested with 2 regularly used commercial ELISAs for flavivirus screening in animals (Immunozym FSME (TBEV) IgG All Species (Progen) and ID Screen West Nile Competition (Innovative Diagnostics)) and PRNTs for TBEV and USUTU virus. The results showed that the Immunozym and IDScreen ELISAs had low relative sensitivities of 23% and 20%, respectively, compared to the PRNT results. The relative specificities were 88% and 84% due to cross reactions with USUTU virus-specific antibodies. The minimal TBEV prevalence in our sample set was 8.6% when determined by PRNT. When the screening approach of ELISA testing followed by PRNT confirmation was applied, a TBEV seroprevalence of only 2.0% and 1.7% was found. The suboptimal performance of the ELISAs was confirmed by testing sera collected from experimentally TBEV-infected sheep. While the PRNT detected TBEV specific antibodies in 94% of samples collected between 7 and 18 days post-infection, the ELISAs classified only 50% and 31% of the samples as positive. Both routinely used ELISAs for TBEV antibody screening in animal sera were shown to have a low sensitivity, potentially leading to an underestimation of the true prevalence, and furthermore cross-react with other flavivirus antibodies.

P816 The efficacy of an over the counter multivitamin and mineral supplement to prevent opportunistic infections in patients with inflammatory bowel disease in remission

Abstract Background Patients with inflammatory bowel disease (IBD) treated with immunomodulators or biological therapy are at increased risk of infections. Malnutrition and vitamin or mineral deficiencies are common among patients with IBD. The results of various studies have indicate that vitamin deficiencies might increase the risk of infections. Methods Single-centre, randomized, double-blinded, placebo-controlled clinical trial to compare a multivitamin and mineral supplement (vitamin group) versus identical in appearance placebo (placebo group). Patients with Crohn’s disease or ulcerative colitis using immunomodulators and/or biological therapy without vitamin deficiency were considered for participation in our study. Participants were asked to take a daily multivitamin and mineral supplement or placebo and report the occurrence of infections during a 24 weeks period of follow up. The primary endpoint of this study was the difference in incidence of infection between the two groups measured by a validated questionnaire, the revised version of the ID screen to assess infection13. Randomization was conducted by using computer generated random numbers with an allocation ratio of 1:1 to vitamin supplement or placebo in blocks of 8 and stratified according to type of IBD.Results Between July 2020 and February 2021, 320 patients were randomized. Treatment arms consisted out of 162 and 158 for vitamin and placebo, respectively. Both groups showed similarity in patient characteristics. All patients were included in the intention-to -Treat (ITT), 265 patients in the per protocol (PP) (compliance >80%) analyses. For both groups, 107 patients reported an infection during the 24 weeks follow up period (Unadjusted Odds Ratio: 0.93 (95% confidence interval: 0.56-1.48). In the vitamin group, 32 patients received antibiotics for an infection compared with 21 patients in the placebo group (Unadjusted Odds Ratio: 1.61 (95% confidence interval: 0.88-2.93). 166 patients in the PP analysis reported an infection (Unadjusted Odds Ratio: 0.91 (95% confidence interval: 0.55-1.50), 23 reported a serious infection (Unadjusted Odds Ratio: 2.57 (95% confidence interval: 1.02 -6.46). None of the patients was hospitalized due to serious infection. Conclusion An over the counter multivitamin and mineral supplement did not reduce the risk of infection for patients with IBD in remission with immunomodulators and/or biological therapy.

Seroprevalence and associated factors of infectious bursal disease (IBD) in indigenous chicken in eastern province in Rwanda

ABSTRACT The status of Infectious bursal disease (IBD) in indigenous chickens and backyard poultry in Rwanda has not been previously elucidated. This cross-sectional study was to determine the seroprevalence of infectious bursal disease in indigenous chickens and to identify the associated factors. The study was been done in three districts in the Eastern province of Rwanda where blood from 364 indigenous chickens were collected. ID Screen® IBD indirect enzyme-linked immunosorbent assay (ELISA) test was used to detect IBD antibodies in these birds. 145 questionnaires were also administered to poultry farmers to obtain information on biosecurity measures and associated factors to IBD outbreaks. The study revealed 48.4% (176/364) prevalence of the chicken with IBDV antibodies with statistical significance (P < .05) among/between location and age groups. The questionnaire revealed that there were other important associated factors which included chicken scavenging for seed as a source of food (59.3% of farmers reported), absence of routine vaccination (53.8%), live chickens are purchased from the open market with no information about IBD outbreaks and vaccination (30.0%), open disposal of dead chickens suspected of IBD (58.9%). IBD virus antibodies are present in indigenous chicken in Eastern Rwanda hence further investigation to better understand the epidemiology of IBD virus in indigenous chickens is desired and more research is needed to identify the role of indigenous chickens in the spread of IBD virus in Rwanda.

Serological Determination of West Nile Virus in Domestic Birds from Rapa Nui, Chile.

Background: Flaviviruses are agents with high zoonotic potential of importance to human health. They are transmitted by mosquitoes of the Culicidae family, and birds act as host-amplifiers. Birds, mammals, and humans are susceptible hosts to infection. Methods: In this study, West Nile virus (WNV), flavivirus, infection was studied in 37 serum samples from 22 hens on Easter Island, Chile. Results: WNV was detected by ELISA (ID Screen® West Nile Competition Multi-Species). We report absence of antibodies to WNV, and to related viruses of the Japanese Encephalitis Virus serocomplex, and, therefore, absence of infection across the sample. Conclusion: This is the first evaluation of its type carried out in Chile, and represents a positive result for public health at Easter Island.

Prevalence of Bovine Viral Diarrhoea infection in cattle and buffaloes from Andhra Pradesh, India

The present study was directed at assessing the prevalence of Bovine Viral Diarrhoea (BVD) in various districts of Andhra Pradesh. Samples from organized cattle and buffalo farms and also from clinical cases were tested from January 2022 to December 2022. A total of 673 serum samples from five organised cattle and buffalo farms and 86 clinical samples from animals suffering from severe diarrhoea, respiratory problems and reproductive disorders were screened for the presence of antibodies against BVD. Serum samples were tested for antibodies against BVD virus p80-125 protein (NSP2-3) using a commercially available competitive ELISA kit (ID Screen). The results revealed that a total of 442 out of 759 animals (58.23 per cent) were positive for antibodies against Bovine viral diarrhoea virus. Statistically significant association was observed between BVD seropositivity and different risk factors such as district, farm, species and breed. Sample size and Geographical location may have contributed to this result. In two organized farms where seropositivity was high (more than 70%), all the seronegative animals (55) were screened for presence of BVD virus using a commercially available BVD p80 antigen ELISA kit (ID screen) to detect persistently infected animals. As no animal was found positive, all the remaining seropositive animals (414) in these two farms were also tested for BVD antigen on the suspicion of a current transient infection or presence of a seropositive PI due to exposure to a BVDV strain that is antigenically different to the persisting strain. All the animals tested were negative for BVD antigen. Out of all the clinical samples tested, one animal that was antibody positive was found positive for BVD virus also, indicating a transient infection. The high seropositivity in the organized cattle farms without a PI animal might either be due to an earlier transient infection in the herd or transfer of a PI animal away from the herd before testing. The high seropositivity in the state and presence of BVD antigen in the field animal indicates that a circulating BVD virus was present in most parts of the state.

Evidence of West Nile virus in chickens and horses in Nigeria: results from a serosurvey.

West Nile virus (WNV) is an emerging arbovirus which affects humans and horses. A cross sectional study was carried out on 106 local horses in Kaduna and 78 domestic chickens in Federal Capital Territory. A total of 184 sera were screened for West Nile virus anti Pr‑E antibodies using ID Screen® West Nile competitive enzyme linked immunosorbent assay. For the horses, an overall prevalence of 92.45% was recorded while domestic chickens had a preponderance of 7.69%. From our study, there was a statistical significant difference between the occurrences of WNV in stallions than mares with p < 0.05. Comparing the occurrence of West Nile virus between species, horses were more likely to be infected by West Nile virus than domestic chickens (OR 147). This is the first seroprevalence study investigating West Nile virus infection in domestic chickens in Nigeria. The presence of the antibodies indicates the widespread circulation and the potential risk of infection in humans and animals. In order to understand the epidemiology of West Nile virus infection in Nigeria, there is need for surveillance to be implemented in human and animal sectors.

Detection of SARS-CoV-2 antibodies in a pet dogs and cats living in COVID-19-positive households in Iraq

The goal of this study was to assess SARS-CoV-2 seroprevalence in pet dogs and cats during the coronavirus disease (COVID-19) pandemic in Iraq. Serum samples were collected from 180 animals, including 96 dogs and 84 cats. All animals included in this study were owned animals from a household with at least one COVID-19 patient. Serum samples from dogs and cats were screened for antibodies against SARS-CoV-2 using the commercial ELISA assay (ID Screen® SARS-CoV-2 Double Antigen Multi-species) according to the manufacturer’s instructions. The seroprevalences for SARS-CoV-2 was 7.3% for dogs and 8.3% for cats but this difference was not significant. Also, no significant different in seropositivity between age groups, sex and breeds of pet animals. All positive animals did not show any clinical signs when contact with human SARS-CoV2 positive cases, suggesting that dogs and cats are likely asymptomatic carriers of SARS-CoV-2.

Detection of Crimean-Congo Hemorrhagic Fever Virus Antibodies in Cattle in Plateau State, Nigeria

Crimean-Congo hemorrhagic fever (CCHF) is a vector-borne viral hemorrhagic disease with global clinical significance. Certain species of ticks are vectors of CCHF, which can be transmitted from animals to humans and humans to humans by direct exposure to blood or other body fluids. The zoonotic transmission at the human-animal interface from viremic animal hosts to humans is a public health concern with a paucity of data in Nigeria. Samples from 184 pastoral cattle from three local government areas (LGAs) of Plateau state, Nigeria, were screened for CCHF virus using a commercial enzyme-linked immunosorbent assay (ID Screen® CCHF Double Antigen for Multi-Species). Overall seropositivity of 30.4% (n = 56) (95% CI: 23.88%, 37.63%) was recorded from the study areas in Plateau State, while 48/126 (38.1%, 95% CI: 29.59%, 47.17%) sampled cows tested positive for CCHFV antibodies. Seropositivity was significantly higher (p < 0.001) among older cattle greater than two years, 54.69% (95% CI: 2.88%, 11.24%) compared to cattle younger than two years, 17.5% (95% CI: 11.17%, 25.50%). The location of farms played a significant role in the seropositivity of CCHF with the least risk observed in Wase LGA. CCHF is an important zoonotic disease in different parts of the globe with a high risk of transmission to pastoralists, livestock keepers/slaughterhouse workers, and veterinarians who handle animals. There is a need for a collaborative one-health approach with various stakeholders to unravel the dynamics of CCHFV epidemiology in Nigeria.

Comparison of diagnostic performances of different serological tests for SARS-CoV-2 antibody detection in cats and dogs.

SummarySerosurveillance among animals, including pets, plays an important role in the current coronavirus disease 2019 (COVID‐19) pandemic, because severe acute respiratory coronavirus 2 (SARS‐CoV‐2) infections in animal populations could result in the establishment of new virus reservoirs. Serological assays that offer the required sensitivity and specificity are essential. In this study we evaluated the diagnostic performance of three different commercially available immunoassays for the detection of SARS‐CoV‐2 antibodies in pets, namely two ELISA tests for the detection of antibodies against SARS‐CoV‐2 nucleocapsid [ID Screen SARS CoV‐2 double antigen multispecies (Double antigen) and ID Screen® SARS‐CoV‐2‐N IgG indirect ELISA (Indirect)] and one test for the detection of neutralizing antibodies against SARS‐CoV‐2 receptor‐binding‐domain [surrogate virus neutralization test (sVNT)]. The obtained results were compared with those of conventional virus neutralization test (VNT), which was regarded as reference method. A total of 191 serum samples were analyzed. Thirteen (6.8%) samples showed VNT positive results. The overall sensitivity was higher for sVNT (100%) compared to nucleocapsid‐based ELISA assays (23% for Double antigen and 60% for Indirect). The specificity was 100% for Indirect ELISA and sVNT, when a higher cut‐off (>30%) was used compared to the one previously defined by the manufacturer (>20%), whereas the other test showed lower value (99%). The sVNT test showed the highest accuracy and agreement with VNT, with a perfect agreement when the higher cut‐off was applied. The agreement between each nucleocapsid‐based ELISA test and VNT was 96% for Indirect and 94% for Double antigen. Our findings showed that some commercially available serological tests may lead to a high rate of false negative results, highlighting the importance of assays validation for the detection of SARS‐CoV‐2 antibodies in domestic animals.This article is protected by copyright. All rights reserved

Evaluation of three commercial ELISA tests for serological detection of maedi-visna virus using Bayesian latent class analysis

Early and accurate diagnosis is fundamental for successful surveillance and control of maedi-visna virus (MVV). MVV was detected in Norway in 2019, almost 14 years after the previous outbreak. Genetic analysis indicates persistence of the virus in the sheep population since 2005. The virus was not detected despite continuous serological surveillance. This emphasises the need for improved surveillance, which relies on an understanding of both diagnostic test performance, sampling strategy and the prevalence of the disease. This study therefore aims to evaluate three commercial ELISA tests for MVV antibodies. We conducted a retrospective study using 615 samples from six flocks diagnosed with MVV in 2019. We ran all samples with the following three tests: ID Screen® MVV/CAEV Indirect (IDvet, Grabels, France), IDEXX MVV/CAEV p28 Ab Verification Test (IDEXX Laboratories, Maine, USA) and Elitest MVV/CAEV (Hyphen Biomed, Neuville-sur-Oise, France), hereinafter referred to as ID Screen, IDEXXp28 and Elitest respectively. Without a perfect reference test, we used Bayesian latent class analysis, including conditional dependence between tests, to estimate diagnostic accuracy and true prevalence in the flocks. Using recommended cut-off values, we found that ID Screen and Elitest had significantly higher sensitivity (Se) estimates (99.3 % [97.4–100.0, 95 % Posterior Credible Interval] and 97.4 % [94.1–99.7 %], respectively) than IDEXXp28 (79.5 % [72.3–86.0 %]), while IDEXXp28 and ID Screen had significantly higher specificity (Sp) estimates than Elitest (99.7 % [99.1–100.0], 99.1 % [98.0–99.8 %] and 93.7 % [91.4–95.7 %], respectively). The estimated true prevalence in the six flocks ranged from a median of 0.8–93.5 %. Combining ID Screen and Elitest in serial interpretation showed the highest median Se and Sp (96.7 % [92.0–99.1] and 100.0 % [99.9–100.0], respectively), as well as the highest median positive predictive value (PPV) for the population with the lowest prevalence. Our study supports the use of ID Screen for screening. Further verification with Elitest in serial interpretation will enhance the PPV.

Herd-level prevalence of Mycoplasma bovis in Swedish dairy herds determined by antibody ELISA and PCR on bulk tank milk and herd characteristics associated with seropositivity.

Mycoplasma bovis is an important pathogen causing pneumonia, mastitis, and arthritis in cattle, leading to reduced animal welfare and economic losses worldwide. In this cross-sectional study, we investigated the prevalence of M. bovis in bulk tank milk (BTM) and herd characteristics associated with a positive antibody test result in Swedish dairy herds. Bulk tank milk samples from all Swedish dairy herds (n = 3,144) were collected and analyzed with ID Screen antibody ELISA and PCR. Information on herd characteristics was collected from the national Dairy Herd Improvement database. To identify herd characteristics associated with the presence of antibodies in BTM, logistic regression was used in 4 different models. The apparent herd-level prevalence of M. bovis infection based on antibodies in BTM was 4.8%, with large regional differences ranging from 0 to 20%. None of the BTM samples was positive by PCR. All the antibody-positive herds were situated in the south of Sweden. The logistic regression model showed that larger herds had higher odds of detectable antibodies in BTM (herd size >120 cows, odds ratio = 8.8). An association was also found between antibodies in BTM and both a higher late calf mortality (2-6 mo) and a higher young stock mortality (6-15 mo). This study showed a clear regional difference in the apparent prevalence of M. bovis infection based on antibodies. The relatively low prevalence of M. bovis in Sweden is a strong motivator for the cattle industry to take steps to prevent further spread of the infection. It is essential that the M. bovis status of free herds be known, and the regional differences shown in this study suggest that testing is highly recommended when live cattle from high-prevalence areas are being introduced into herds. We do not recommend using PCR on BTM to detect infected herds, owing to the low detection frequency in this study.

SEROPREVALENCE OF CHLAMYDOPHILA ABORTUS IN SHEEP IN ТНЕ BELGRADE EPIZOOTIOLOGICAL AREA DURING 2019-2021

As one of the most important infective agents of abortion, Chlamydophila abortus takes an important place in pathology of ruminant reproductive tract. In sheep, the disease known as enzootic abortion of ewes or ovine enzootic abortion, and is manifested as abortion and accompanied reproductive disorders, thus resulting in significant economic losses worldwide. The characteristics of the pathogen and its zoonotic potential make this disease significant both for animal and public health. Therefore, the presented study aimed to obtain Chlamydophila abortus seroprevalence in sheep population in Belgrade epizootiological area. The study was done by testing 552 sheep sera samples from 10 municipalities of Belgrade city, during the 2019-2021 period. Serological examination was performed using ELISA assay (ID Screen® Chlamydophila abortus Indirect Multi-species, IDvet, Grabels, France). The obtained results showed Chlamydophila abortus seroprevalence of 6% in Belgrade epizootiological area. The largest number of seropositive sheep was found in municipality of Palilula. The detected antibodies against Chlamydophila abortus confirmed the circulation of the pathogen in sheep population in Belgrade epizootiological area. The obtained results show the need for further studies and continuous implementation of measures for detection, control, prevention and eradication of the disease.

Seroprevalence and risk factors associated with maedi visna virus in sheep population in southwestern Croatia

The objective of this cross-sectional study was to detect the presence of maedi visna virus (MVV) infections in sheep in three counties in southwestern Croatia: Istria County, Primorje-Gorski Kotar County and Lika-Senj County, and to estimate the influence of geographical location, breed, age and gender as risk factors on the prevalence rate. A total of 460 randomly selected sheep were tested using a commercially available ID Screen® MVV/CAEV Indirect screening test, VISNAS-5P (ID.VET, Grabels France) for detection of antibodies against the Maedi-Visna virus. The prevalence rate for the entire population was 10.00 % (95 % 7.26 - 12.74 %), The highest seroprevalence of MVV was found in Istria County (19.02%, CI 95% 13.35 –24.70%), followed by Lika-Senj County (5.43 %, CI 95 % 2.17- 8.70) and Primorje-Gorski Kotar County (1.07 %, CI 95 % -1.03 – 3.17) (P&lt;0.01). When comparing the seroprevalence among breeds, statistically significant differences were observed. The largest number of positive sheep was found in the following breeds: Istrian Pramenka (22.97%, CI 95% 13.39 - 32.55%), Jezersko-Solcava sheep (23.53%, CI 95% 11.89 - 35.17%%), Pramenka (13.16%, CI 95% 2.41 - 23.91%) and Lika sheep (9.78%, CI 95% 3.71- 15.85%). Prevalence rates in other examined breeds (Krk sheep, Pag sheep and Romanov sheep) were lower than 2 % (P&lt;0.01). No statistically significant differences in seroprevalence between sex and age were found. According to risk estimation, the main risk factors were found to be geographical location and breed. In Istria County, seroprevalence was 4.77 ( CI 95 % 2.67 – 8.54) higher than in other examined areas (OR = 5.66, CI 95 % 2.97-10.79) (P&lt;0.01). As for breeds, significant differences were observed between Jezersko-Solcava sheep and Lika sheep (OR = 2.84, CI 95% 1.13-7.14) and Istrian Pramenka in relation to Pag sheep (OR 22.51,, CI 95% 16.99-119.73), Krk sheep (OR 19.53, CI 95% 5.80-108.27), and Lika sheep (OR 2.35, CI 95% 1.17-6.47). This cross-section study indicates the circulation of MVV in this sheep population in Croatia, and represents the first research into the risk factors for MVV in sheep in southwestern Croatia.

Prevalence of Mycoplasma bovis in Algeria and Characterisation of the Isolated Clones.

Bovine respiratory disease (BRD) is common in calves in Algeria, but to date, Mycoplasma bovis has never been monitored as a potential etiological agent. Here, to assess the presence (direct detection) and circulation (indirect detection) of M. bovis, broncho-alveolar lavage fluids (BALF) and serum samples were collected from 60 veal calf farms in Algeria. A commercial ELISA kit (ID Screen® ELISA) was used to screen for the presence of specific antibodies against M. bovis in 351 blood sera collected from both diseased and healthy calves, and 69% (241 sera) tested positive. BALFs from the 176 diseased calves were used to screen for M. bovis by real-time-PCR (rt-PCR), and 102 (58%) tested positive. A non-exhaustive set of 53 clones were isolated from 44 calves and further subtyped using polC gene sequencing. No predominant subtype was found, and two clones exhibited a new subtype. Fourteen clones were further characterized by multilocus sequence typing, and results showed a high degree of genetic diversity, with some clones having new alleles and subtypes. The minimum inhibitory concentrations (MICs) of 5 antimicrobials regularly used to treat BRD was determined on 45 clones. Susceptibility profiles showed very broad diversity, confirming the variety of clones actively circulating. We detected clones with high MICs, including increased MICs of enrofloxacin (n = 5). This is the first study to report the presence of M. bovis in Algeria in calves with BRD. This research also finds broad genetic and phenotypic diversity in the actively circulating isolates.

Development of an ELISA to distinguish between foot-and-mouth disease virus infected and vaccinated animals utilising the viral non-structural protein 3ABC.

Introduction. Foot-and-mouth disease (FMD) is a highly contagious and economically devastating viral disease of livestock and is endemic in much of Asia, including Pakistan. Vaccination is used to control disease outbreaks and sensitive diagnostic methods which can differentiate infected animals from vaccinated animals (DIVA) are essential for monitoring the effectiveness of disease control programmes. Tests based on the detection of the non-structural protein (NSP) 3ABC are reliable indicators of virus replication in infected and vaccinated populations.Hypothesis/Gap statement. Diagnosis of FMD is expensive using commercial ELISA kits, yet is essential for controlling this economically-important disease.Aim. The development of a low-cost diagnostic ELISA, using protein made in Escherichia coli.Methodology. In this study, the viral precursor protein 3ABC (r3ABC) was expressed in E. coli, solubilised using detergent and purified using nickel affinity chromatography. The fusion protein contained an attenuating mutation in the protease and a SUMO tag. It was characterised by immunoblotting and immunoprecipitation, which revealed antigenicity against virus-specific polyclonal sera. Using r3ABC, an indirect ELISA was developed and evaluated using field sera from healthy/naïve, vaccinated and infected animals.Results. The diagnostic sensitivity and specificity of the r3ABC in-house ELISA were 95.3 and 96.3% respectively. The ELISA was validated through comparison with the commercially available ID Screen FMD NSP competition kit. Results indicated good concordance rates on tested samples and high agreement between the two tests.Conclusion. The ELISA described here can effectively differentiate between infected and vaccinated animals and represents an important low cost tool for sero-surveillance and control of FMD in endemic settings.

Seroprevalence of Mycoplasma bovis in bulk milk samples in Irish dairy herds and risk factors associated with herd seropositive status

Mycoplasma bovis is a serious disease of cattle worldwide; mastitis, pneumonia, and arthritis are particularly important clinical presentations in dairy herds. Mycoplasma bovis was first identified in Ireland in 1994, and the reporting of Mycoplasma-associated disease has substantially increased over the last 5 years. Despite the presumed endemic nature of M. bovis in Ireland, there is a paucity of data on the prevalence of infection, and the effect of this disease on the dairy industry. The aim of this observational study was to estimate apparent herd prevalence for M. bovis in Irish dairy herds using routinely collected bulk milk surveillance samples and to assess risk factors for herd seropositivity. In autumn 2018, 1,500 herds out of the 16,858 herds that submitted bulk tank milk (BTM) samples to the Department of Agriculture testing laboratory for routine surveillance were randomly selected for further testing. A final data set of 1,313 sampled herds with a BTM ELISA result were used for the analysis. Testing was conducted using an indirect ELISA kit (ID Screen Mycoplasma bovis). Herd-level risk factors were used as explanatory variables to determine potential risk factors associated with positive herd status (reflecting past or current exposure to M. bovis). A total of 588 of the 1,313 BTM samples were positive to M. bovis, providing an apparent herd prevalence of 0.45 (95% CI: 0.42, 0.47) in Irish dairy herds in autumn 2018. Multivariable analysis was conducted using logistic regression. The final model identified herd size, the number of neighboring farms, in-degree and county as statistically significant risk factors for herd BTM seropositivity to M. bovis. The results suggest a high apparent herd prevalence of seropositivity to M. bovis, and evidence that M. bovis infection is now endemic in the Irish dairy sector. In addition, risk factors identified are closely aligned to what we would expect of an infectious disease. Awareness raising and education about this important disease is warranted given the widespread nature of exposure and likely infection in Irish herds. Further work on the validation of diagnostic tests for herd-level diagnosis should be undertaken as a matter of priority.

Serosurvey of SARS-CoV-2 in dogs and cats from Portugal

PurposeSevere acute respiratory syndrome–coronavirus 2 (SARS-CoV-2) causes COVID-19, which was first reported in humans in 2019, in Wuhan, China. This RNA virus has highly efficient transmission. Sporadic cases of infection in pets have been described. Human to animal transmission seems to occur, however, the epidemiologic role of pets remains unclear. This study aimed to screen dogs and cats from the North and Centre regions of Portugal for the exposure to SARS-CoV-2, during the peak of SARS-CoV-2 human infection in Portugal, which occurred between October 2020 and March 2021.Methods & MaterialsA cross-sectional study was conducted in dogs and cats presented to veterinary medical centres from Portugal between October 2020 and March 2021, that required blood sampling as part of their diagnostic plan. Only surplus sera or plasma samples were used in this research. Sera or plasma were tested for the presence of specific antibodies anti-SARS-CoV-2 using a commercial ELISA adapted for multi-species detection (ID Screen SARS-CoV-2Double Antigen Multispecies®, IDVet). Laboratory results were expressed in S/P values and samples with an S/P% ≥ 60% were classified as positive.ResultsA total of 107 animals were sampled (dogs n=60; cats n=47). The canine population was composed by 25 purebred and 35 crossbreed dogs, with ages ranging from 5 months to 15 years of age. Cats were mainly of the domestic short-hair breed (n=45), with ages ranging between 6 months and 9 years old. The estimated rate of exposure was of 5.0% (95% CI: 1.71-13.7%) in dogs (n=3) and 2.13% (95% CI: 1.18-14.26%) in cats (n=1). A doubtful result (50≥S/P%<60%) was obtained in 6.7% (95% CI: 2.6-15.9%) of dogs (n=4) and in 4.26% (95% CI: 2.6-15.9%) of cats (n=3).ConclusionTo our best knowledge, this is the first serosurvey conducted in pets in Portugal. An exposure to the agent has been evidenced in dogs and in cats. Further studies must clarify the impact of the exposure in animal health and the role of the pets in spreading the virus.

Coxielosis in dairy cattle herds: serological and molecular study

<h3>Purpose</h3> Ruminants are recognized as the main reservoirs of <i>Coxiella burnetii</i>. In Portugal, in 2014, in a survey in apparently healthy dairy cattle herds from the center region, 37.8% of herds evidenced an exposure to <i>C. burnetii</i>. Furthermore, milk shedding was demonstrated in 20% of these herds. The present study aimed: i) to investigate the exposure to <i>C. burnetii</i> in dairy cattle herds from the same region by testing BTM to the presence of specific anti-<i>C. burnetii</i> antibodies ii) to evaluate the milk shedding in exposed herds iii) to evaluate the degree of bacterial excretion in shedding herds; iv) compare results with those obtained in 2014(1). <h3>Methods & Materials</h3> A cross-sectional study was carried out in dairy cattle herds (n=30) from central Portugal with no previous vaccination to Q fever. Bulk tank milk (BTM) samples were collected and tested by a commercial ELISA (ID Screen Q Fever Indirect Multispecies®, IDVet). Antibodies positive BTM samples were tested by qPCR (Taq-Vet™ Coxiella burnetii–Absolute Quantification, Thermofischer®) <h3>Results</h3> A total of 5398 females with a mean age of 3.2 years (SD=0.73) from 30 herds were tested. An antibody positive result was obtained in 96.7% (95% CI: 83.3-99.4) of herds and S/P% ranged from 49 to 432. The shedding of <i>C. burnetii</i> in BTM was detected in 31.03% (95% IC: 17.27-49.23) and a high bacterial load (≥3 × 10³ bacteria/mL) was observed in 44.4% of PCR positive BTM. Comparison of results from 2014 to 2021, shows that the exposure to <i>C. burnetii</i> increased significantly (p<0,0001). <h3>Conclusion</h3> Serological and molecular methods in combination are a crucial tool for Coxielosis surveillance. From 2014 until 2021, an increase of the exposure as well as an increase of bacteria shedding in BTM was observed. It is recognized that the shedding of <i>C. burnetii</i> through milk is important, especially in dairy cattle, and thus the milk as a potential source of infection among dairy workers should not be neglected.

Detection of Anti-Neospora caninum Antibodies on Dairy Cattle Farms in Southern Italy.

Neosporosis is recognized as one of the major causes of bovine abortion worldwide. Canids are the main definitive host for this parasite and the presence of dogs in the farm is an important factor for the Neospora caninum infection in bovines. Since, in the province of Lecce, located in the Apulia region of Southern Italy, there are no studies showing the presence of the infection in farm animals, the objective was to perform a serological evaluation for anti-N. caninum antibodiesin serum from 706 dairy cattle and 21 farm dogs located in 40 farms uniformlydistributed over the territory.The presence of N. caninum infection was confirmed in 90.0% (36/40) of the 40 farms examined. The results obtained on all serum samples by an enzyme-linked immunosorbent assay (ID Screen® Neospora caninum competition ELISA kit) for anti-N. caninum antibodies showed a seropositivity rate of 21.1% (149/706) among dairy cows, with a statistically significant higher percentage of positive subjects in the animals over two years old and a positivity rate of 42.9% (9/21) in tested dogs. The obtained data confirmed the presence of neosporosis even in the Lecce area, where it could therefore represent an important cause of abortion and economic losses.