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Human echinococcosis is present worldwide but it is in China that disease prevalence is the highest. In western China, especially in the Tibetan Plateau, the burden of echinococcosis is the most important. Dogs are a major definitive host of Echinococcus and monitoring the presence of Echinococcus worms in dogs is therefore essential to efficiently control the disease. Detection kits based on three different technologies including sandwich ELISA, (indirect) ELISA, and gold immunodiffusion, are currently marketed and used in China. The objective of this work was to assess the efficacy of these kits, in particular with respect to sensitivity and specificity. Four fecal antigen detection kits for canine infection reflecting the three technologies were obtained from companies and tested in parallel on 220 fecal samples. The results indicate that the performance is lower than expected, in particular in terms of sensitivity. The best results were obtained with the sandwich ELISA technology. The gold immunofiltration yielded the poorest results. In all cases, further development is needed to improve the performance of these kits which are key components for the control of echinococcosis., Author summary Although present worldwide, human echinococcosis is at its highest prevalence in western China and particularly in the Tibetan Plateau. Controlling echinococcosis is a national priority and routine monitoring must be established. Dogs are the main infection source in China and surveying Echinococcus worms in dogs is therefore a research priority. Commercial detection kits are currently in use in China to monitor the presence of Echinococcus in dogs. The kits are based on three different technologies including sandwich ELISA with two monoclonal antibodies, (indirect) ELISA, and gold immunodiffusion. National survey programmes are essential for the control of echinococcosis and it is thus very important to assess the efficacy of these kits for the programmes. This work was therefore undertaken to assess efficacy of the kits, in particular with respect to sensitivity and specificity. Four fecal antigen detection kits for canine infection reflecting the three technologies were obtained from companies and tested in parallel on 220 fecal samples. The performance was lower than expected, in particular for their sensitivity which ranged between 51.5% and 83.9% with only two samples displaying a worm burden lower than 100. Three out of four kits showed non-specific cross-reactions with other parasites. The best results were obtained with the sandwich ELISA technology, whereas gold immunofiltration yielded the poorest results. However, in all cases, further development is strongly needed to improve the performance of these kits which are key components for the control of echinococcosis.

Animal trypanosomoses due to trypanosomes of African origin (ATAO), mainly caused by Trypanosoma congolense type Savannah (TCS), T. brucei brucei (TBB), T. vivax (TV), and T. evansi, are widespread diseases that affect domestic and wild mammals and have a huge economic impact. ATAO clinical suspicions are usually confirmed by parasitological and molecular methods, while sero-epidemiological surveys are generally carried out using the OIE-recommended ELISA method based on whole cell lysate soluble antigens (WCLSA) from purified trypanosomes; this reagent is usually stored frozen. With a view to expanding this ELISA test, we assessed, standardized, and validated the use of dehydrated rather than frozen WCLSA and serum samples. For the three ELISA assays (TV, TCS & TBB), a repeatability study revealed no significant difference between repeats. The results obtained using frozen rather than freeze-dried antigen and serum strongly correlated for Pearson’s correlation values (>0.93) and Lin’s measure (“very good” to “excellent”). Reproducibility was robust, with Pearson’s correlation values >0.97 for inter technician effects, and 0.87 (TV) to 0.97 (TBB & TCS) for inter-laboratory tests; their combination was “very satisfactory” to “excellent” according to Lin’s measure and there was no impact on qualitative test results. Dehydrated reagents offer the advantage of shipment at room temperature, allowing the secured provision of reagents to regional laboratories. Together with a compendium of standard diagnostic protocols for ATAO (/OIE), dehydrated reagents will enable the serological diagnosis of ATAO at regional level in endemic countries. This very welcome improvement in the context of the Progressive Control Pathway for trypanosomes, recently launched by African countries, will possibly be extended to Latin America in the near future.

Cypress (Cupressus sp.pl) is a genus within the Cupressaceae family. This family covers all of the Earth's continents except for Antarctica, and it includes about 160 species. The most important taxa for allergic diseases belong to five different genera: Cupressus, Hesperocyparis, Juniperus, Cryptomeria, and Chamaecyparis. Cupressaceae species share a common pollen type that can even include the genus Taxus (Taxaceae) when this plant is also present. As Juniperus oxycedrus pollinates in October, Cupressus sempervirens in January and February, Hesperocyparis arizonica (prev. Cupressus arizonica) in February and March, and Juniperus communis in April, the symptomatic period is long-lasting. Due to global warming, the pollination period tends to last longer, and there is a trend for Cupressaceae bioclimate niches to migrate north. In Mediterranean areas, C. sempervirens (Italian cypress or Mediterranean cypress) is by far the most common pollinating species. It accounts for half of the total pollination level. The group 1 major allergens belong to the pectate-lyase family, and members share 70 to 97% sequence homology within the different Cupressaceae. Group 2 allergens correspond to the polygalacturonase protein family, while group 3, a minor allergen, belongs to the family of "thaumatin-like proteins," a pathogenesis-related protein 5. Group 4 allergens are Ca++-binding protein (4 EF-hands). Aside from these four groups, about 15 other allergens have been reported. Prominent among these is a basic low-molecular mass cross-reactive allergen that was identified recently, and which is suspected to be involved in pollen food syndromes which are common with peach and citrus. The prevalence of cypress allergy in the general population ranges from 0.6 to 3%, depending on the degree of exposure to the pollen. Depending on the geographic area and the studied population, 9 to 65% of outpatients consulting an allergist may have sensitization to cypress pollen. Repeated cross-sectional studies performed at different time intervals have demonstrated a threefold increase in the percentage of cypress allergy around the Mediterranean area. Risk factors include a genetic predisposition and/or a strong exposure to pollen, and the natural history of cypress allergy allows identification of a subgroup of patients as allergic rather than atopic. Concerning the clinical expression, rhinitis is the most prevalent symptom, while conjunctivitis is the most disabling. Pharmacological treatment of cypress allergies is not different from that of other seasonal allergies. Immunotherapy has been used, initially by subcutaneous injections, but currently mostly through the sublingual route. Although clinical trials have included only a limited number of patients, it has proven effective and safe. Avoidance can be implemented at the individual level, as well as at the community level, through the use of alternative plants, low-pollinating cypresses, or by trimming hedges before pollination.

Contagious bo vine pleuropneumonia (CBPP) is a disease of cattle caused by Mycoplasma mycoides subsp. mycoides SC (MmmSC; SC = small colonies). lt is manifested by anorexia, fever and respiratory signs such as dyspnoea, polypnoea, cough and nasal discharges in bovines. Diagnosis depends on the isolation of the aetiological agent. The main problems for control or eradication are the frequent occurrence of subacute or subclinical infections and the persistence of chronic carriers after the clinical phase. Identification of the agent: Samples to be taken from live animals are nasal swabs and/or broncho-alveolar washings or pleural fluid obtained by puncture. Samples to be taken at necropsy are lung lesions, lymph nodes, pleural fluid and synovial fluid from those animals with arthritis. Direct examination of the exudate or smears is possible, but requires great skill. For cultivation of the pathogen, the tissues are ground in medium containing antibiotics and inoculated into media that contain inhibitors to prevent the growth of contaminating bacteria. The growth of MmmSC takes several days. In broth, growth is visible within 2-4 days as a homogeneous cloudiness with whirls when shaken; on agar, small colonies develop, 1 mm in diameter, with the classical 'fried-egg' appearance. The biochemical characteristics of MmmSC are the following: sensitivity to digitonin, reduction of tetrazolium salts, fermentation of glucose, absence of arginine hydrolysis, and the absence of or very slight phosphatase and proteolytic activities. Special media have been described that are recommended for these tests. Diagnosis is confirmed by immunological tests, such as the growth inhibition and immunofluorescence tests (both use hyperimmune sera). The polymerase chain reaction is now used as a rapid, specific, sensitive and easy to use test. Serological tests: For diagnosis, the modified Campbell & Turner complement fixation test remains a prescribed test for international trade. However, it has significant limitations regarding sensitivity and specificity. The competitive enzyme-linked immunosorbent assay was designated as an OIE prescribed test for international trade by the OIE International Committee in May 2004. An immunoblotting test has undergone evaluation and is highly specific and sensitive. Requirements for vaccines: A ttenuated strains now recommended for vaccine production: the T1/44 and T1sr. The minimal recommended titre is 107 mycoplasmas per vaccinal dose, but higher titres of at least 1 oB are recommended.

Background WHO developed a global strategy to eliminate hepatitis B by 2030 and set target to treat 80% of people with chronic hepatitis B virus (HBV) infection eligible for antiviral treatment. As a first step to achieve this goal, it is essential to conduct a situation analysis that is fundamental to designing national hepatitis plans. We therefore estimated the prevalence of chronic HBV infection, and described the existing infrastructure for HBV diagnosis in Madagascar. Methods We conducted a stratified multi-stage serosurvey of hepatitis B surface antigen (HBsAg) in adults aged ≥18 years using 28 sentinel surveillance sites located throughout the country. We obtained the list of facilities performing HBV testing from the Ministry of Health, and contacted the person responsible at each facility. Results A total of 1778 adults were recruited from the 28 study areas. The overall weighted seroprevalence of HBsAg was 6.9% (95% CI: 5.6–8.6). Populations with a low socio-economic status and those living in rural areas had a significantly higher seroprevalence of HBsAg. The ratio of facilities equipped to perform HBsAg tests per 100,000 inhabitants was 1.02 in the capital city of Antananarivo and 0.21 outside the capital. There were no facilities with the capacity to perform HBV DNA testing or transient elastography to measure liver fibrosis. There are only five hepatologists in Madagascar. Conclusion Madagascar has a high-intermediate level of endemicity for HBV infection with a severely limited capacity for its diagnosis and treatment. Higher HBsAg prevalence in rural or underprivileged populations underlines the importance of a public health approach to decentralize the management of chronic HBV carriers in Madagascar by using simple and low-cost diagnostic tools. Electronic supplementary material The online version of this article (doi:10.1186/s12889-017-4630-z) contains supplementary material, which is available to authorized users.

Trypanosoma congolense and T. vivax are the main causative agents of animal African trypanosomosis (AAT), a disease which hinders livestock production throughout sub-Saharan Africa and in some parts of South America. Although two trypanocidal drugs are currently available, the level of treatment is low due to the difficulty in diagnosing the disease in the field. The major clinical signs of AAT such as anaemia, weight loss, and infertility, are common to several other endemic livestock diseases. Current diagnostic methods, based on the visualization of the parasite in the blood, or on the detection of its DNA or the antibodies it triggers in the host, are not suitable for direct use in the field as they require specialized equipment and personnel. Thus, we developed a quick-format diagnostic test (15 min) based on the recombinant TcoCB and TvGM6 antigens for detection of T. congolense and T. vivax , respectively, aimed at providing farmers and veterinarians in the field with the means to conduct a quick diagnosis. The specificity and sensitivity of the test were evaluated using sera from experimentally infected cattle, and fresh blood when possible. The prototype, which includes both antigens, shows a specificity of 95.9 (95% C.I., 90.4%–100%) and a sensitivity of 92.0% (95% C.I., 85.9%–98.1%) for T. congolense and 98.2% (95% C.I., 94.7%–100%) for T. vivax . The high levels of sensitivity and specificity of this rapid test, the possibility of using directly whole blood, and the ease of interpreting the result, all contribute to make of this test a valuable candidate to contribute to the control of AAT in the field. However, further tests with more representative, numerous and fresh reference samples are necessary in order to compare this test with the ELISA, the current gold standard serological test for trypanosomosis.

From May to September 2013, monthly samples were collected from swine in a Vietnamese slaughterhouse for influenza virus isolation and serological testing. A(H1N1)pdm09 viruses and a novel H3N2 originating from reassortment between A(H1N1)pdm09 and novel viruses of the North American triple reassortant lineage were isolated. Serological results showed low seroprevalence for the novel H3N2 virus and higher seroprevalence for A(H1N1)pdm09 viruses. In addition, serology suggested that other swine influenza viruses are also circulating in Vietnamese swine.

International audience; Background: West Nile virus (WNV) is an emergent pathogen that is widely distributed in North and Central America. The recent introduction in South America has focused attention on the spread of WNV across Southern American countries. The transmission network involves mosquitoes, birds, horses and humans. Methods: The serological evaluation of sera from 678 equids and 478 birds was performed using a WNV-specific blocking ELISA, and only the positive results were confirmed by plaque reduction neutralisation tests (PRNTs). Molecular analysis was performed on sera from 992 healthy equids and on 63 macerates of brains from equids that died of encephalitis and had previously tested negative for other pathogens. We also tested swabs from 928 birds. The samples analysed were collected in different biomes of Brazil. Results: We identified WNV antibodies by ELISA in thirteen equids and five birds, and PRNT90 confirmed WNV positivity in four equid samples collected in 2009 in an area between the Amazon and the Pantanal. None of the ELISA positive bird samples were confirmed by PRNT90, and all samples tested by RT-PCR were negative. Conclusion: WNV circulation is confirmed by this large scale survey even in the absence of detection of clinical cases.

African swine fever (ASF) is an infectious disease that causes heavy mortality in domestic pigs. At present there is no vaccine against ASF, and eradication in countries where the disease is endemic is based only on competent diagnosis programs and the sacrifice of infected animals. Due to the presence of natural attenuated strains, certain infection conditions may result in reduced mortality. In these situations, the disease can be diagnosed by detection of specific antibodies. The use of classical and validated diagnosis assays, such as ELISA and Indirect Immunofluorescence or Immunoblotting, allowed the eradication of ASF in the Iberian Peninsula in the 1990s. However, given that conventional tests include the use of antigens obtained from ASF virus (ASFV)-infected cells, they have several disadvantages, such as difficulties to achieve standardization and also the risks associated with the manipulation of live virus. Such drawbacks have led to the development of alternative and more robust systems for the production of ASFV antigens for use in anti-ASFV antibody detection systems. In the present review, we provide an update on current knowledge about antigen targets for ASFV serodiagnosis, the significant progress made in recombinant antigen production, and the refinement of ASF serological diagnostic assays. Moreover, we describe the accuracy of an ELISA developed for the serodiagnosis of ASFV in Africa. This assay is based on a novel p30 recombinant protein (p30r) obtained from an Eastern African viral isolate (Morara strain), which shares 100% amino acid sequence identity with the Georgia virus isolate. That study included the analyses of 587 field sera collected from domestic pigs and warthogs in Senegal (West Africa), the Democratic Republic of Congo (Central Africa), Mozambique (South-East Africa), and South Africa. The results revealed that the novel p30r-based ELISA allows the accurate detection of antibodies against ASFV, independently of the geographical origin of the sera. © 2012 Elsevier B.V.

International audience; Trypanosomosis due to Trypanosoma evansi (surra) is a major enzootic disease of the dromedary camel. The present study was conducted to determine the prevalence of camel trypanosomosis in the northern part of Saudi Arabia with different methods of diagnosis (ELISA, PCR) and to compare the results to whose obtained previously with Card Agglutination Test for Trypanosomiasis (CATT/T.evansi). A total of 195 blood samples and 118 serum samples were used for molecular and serological investigation respectively. After analyses, 25% (49/195) and 3% (4/118) samples were positive using PCR and ELISA respectively. The variability of trypanosomosis was highly significant to the factor moving, location, breed and clinical signs with PCR. The discrepancy between PCR, CATT test and ELISA is likely due to antibodies degradation on spotted papers maintained several weeks at ambient temperature. This is the first molecular diagnosis report which gives a picture of camel trypanosomosis in Al-jouf, Saudi Arabia.

SUMMARYA serosurvey of 349 military working horses and 231 military working dogs was conducted in ten sites in Morocco in 2012. This survey revealed a high level of exposure of these animals to flaviviruses: seroprevalence rates of 60% in horses and of 62% in dogs were observed using a competitive West Nile virus (WNV) enzyme-linked immunosorbent assay (cELISA). Seroneutralization test results showed that the majority of cELISA-positive results were due to exposure to WNV. Further assays conducted in vaccinated horses with a DIVA (Differentiating Infected from Vaccinated Animals) test indicated that anti-WNV antibodies had been stimulated through WNV natural infection. Moreover, in both species, seroneutralization tests suggested an exposure to Usutu virus (USUV). Data analysis did not show any significant difference of cELISA seropositivity risk between horses and dogs. Dogs may thus represent an interesting alternative to equines for the serological surveillance of WNV or USUV circulation, especially in areas where equine vaccination precludes passive surveillance (based on the detection of West Nile fever cases) in horses.

Background The saliva of tsetse flies contains a cocktail of bioactive molecules inducing specific antibody responses in hosts exposed to bites. We have previously shown that an indirect-ELISA test using whole salivary extracts from Glossina morsitans submorsitans was able to discriminate between (i) cattle from tsetse infested and tsetse free areas and (ii) animals experimentally exposed to low or high numbers of tsetse flies. In the present study, our aim was to identify specific salivary synthetic peptides that could be used to develop simple immunoassays to measure cattle exposure to tsetse flies. Methods In a first step, 2D-electrophoresis immunoblotting, using sera from animals exposed to a variety of bloodsucking arthropods, was performed to identify specific salivary proteins recognised in cattle exposed to tsetse bites. Linear epitope prediction software and Blast analysis were then used to design synthetic peptides within the identified salivary proteins. Finally, candidate peptides were tested by indirect-ELISA on serum samples from tsetse infested and tsetse free areas, and from exposure experiments. Results The combined immunoblotting and bioinformatics analyses led to the identification of five peptides carrying putative linear epitopes within two salivary proteins: the tsetse salivary gland protein 1 (Tsal1) and the Salivary Secreted Adenosine (SSA). Of these, two were synthesised and tested further based on the absence of sequence homology with other arthropods or pathogen species. IgG responses to the Tsal152–75 synthetic peptide were shown to be specific of tsetse exposure in both naturally and experimentally exposed hosts. Nevertheless, anti-Tsal152–75 IgG responses were absent in animals exposed to high tsetse biting rates. Conclusions These results suggest that Tsal152–75 specific antibodies represent a biomarker of low cattle exposure to tsetse fly. These results are discussed in the light of the other available tsetse saliva based-immunoassays and in the perspective of developing a simple serological tool for tsetse eradication campaigns to assess the tsetse free status or to detect tsetse reemergence in previously cleared areas. Electronic supplementary material The online version of this article (doi:10.1186/s13071-016-1414-8) contains supplementary material, which is available to authorized users.

International audience; In Vietnam, serological post H5N1 vaccination surveillance using the HI test is applied to assess the efficiency of the vaccination in addition to virological monitoring. In this paper we report on the evaluations of the performances of the haemagglutination inhibition (HI) test and of a H5-ELISA, using chicken and duck field samples. The evaluations were conducted by comparison with a pseudotyped-based virus neutralization test (H5pp VNT) performed in a reference laboratory and considered as a "gold standard" and also by using methods developed for imperfect reference test. Their global accuracy and best cut-offs were also estimated. Results from the HI test for several haemagglutinin subtypes and from a commercial type A influenza competition ELISA were also compared. The results showed that performance of the HI test was very good in comparison with the H5pp VNT. Data also clearly supported the cut-off of ≥4log(2) used for the HI test for chickens but, a 3log(2) positivity cut-off would be more appropriate for ducks. When compared with the VNT, the H5-ELISA showed poor specificity when using the positivity cut-off specified by the manufacturer but could be used as a screening test if confirmed by the HI test or the H5ppVNT which presents some interests for large scale testing (no need for biosafety level 3 conditions and high performance). A general and highly sensitive pre-screening can also be achieved using the detection of NP-specific antibodies with a competition ELISA. This appears of little interest in a context of high subtypes diversity where only a subtype is targeted for surveillance and control.

A Bayesian approach, allowing for conditional dependence between two tests was used to estimate without gold standard the sensitivities of complement fixation test (CFT) and competitive enzyme-linked immunosorbent assay test (cELISA) and the serological prevalence of CBPP in a cattle population of the Central Delta of the Niger River in Mali, where CBPP is enzootic and the true prevalence and animals serological state were unknown. A significant difference (P = 0.99) was observed between the sensitivities of the two tests, estimated at 73.7% (95% probability interval [PI], 63.4-82.7) for cELISA and 42.3% (95% PI, 33.3-53.7) for CFT. Individual-level serological prevalence in the study population was estimated at 14.1% (95% PI, 10.8-16.9). Our results indicate that in enzootic areas, cELISA performs better in terms of sensitivity than CFT. However, negative conditional sensitivity dependence between the two tests was detected, implying that to achieve maximum sensitivity, the two tests should be applied in parallel.

The performance of indirect fluorescence antibody test (IFAT) for serological diagnosis and screening of lumpy skin disease (LSD) was evaluated using methods without gold standard. Virus neutralization test (VNT) was used as the second test and the study sites were selected from two different geographical places in Ethiopia to get different disease prevalence. The analysis of conditional dependent Bayesian model for the accuracy of IFAT showed that sensitivity, specificity, prevalence of the population Pi(1) and the population Pi(2) were 0.92 (0.89-0.95), 0.88 (0.85-0.91), 0.28 (0.25-0.32) and 0.06 (0.048-0.075), respectively. The posterior inferences obtained for VNT sensitivity, specificity and conditional correlation between the tests for sensitivity (rhoD) and specificity (rhoDc) were 0.78 (0.74-0.83), 0.97 (0.95-0.99), 0.052 (-0.03-0.15) and 0.019 (-0.01-0.06), respectively. The interval estimation of conditional correlation for both sensitivity and specificity clusters around zero and thus conditional dependence between the two tests was not significant. Although accuracy measure would not be the only basis for test selection, the result of our study demonstrated that IFAT has a reasonable high accuracy to be used for the diagnosis and sero-surveillance analysis of LSD in the target population.

International audience; Leptospirosis is a zoonosis known to be endemic in the Mekong Delta of Vietnam, even though clinical reports are uncommon. We investigated leptospira infection in rats purchased in food markets during the rainy season (October) (n = 150), as well as those trapped during the dry season (February-March) (n = 125) in the region using RT-PCR for the lipL32 gene, confirmed by 16S rRNA, as well as by the microscopic agglutination test (MAT). Results were compared with the serovar distribution of human cases referred from Ho Chi Minh City hospitals (2004-2012) confirmed by MAT (n = 45). The MAT seroprevalence among rats was 18.3%. The highest MAT seroprevalence corresponded, in decreasing order, to: Rattus norvegicus (33.0%), Bandicota indica (26.5%), Rattus tanezumi (24.6%), Rattus exulans (14.3%), and Rattus argentiventer (7.1%). The most prevalent serovars were, in descending order: Javanica (4.6% rats), Lousiana (4.2%), Copenageni (4.2%), Cynopterie (3.7%), Pomona (2.9%), and Icterohaemorrhagiae (2.5%). A total of 16 rats (5.8%) tested positive by RT-PCR. Overall, larger rats tended to have a higher prevalence of detection. There was considerable agreement between MAT and PCR (kappa = 0.28 [0.07-0.49]), although significantly more rats were positive by MAT (McNemar 29.9 (p < 0.001). MAT prevalence was higher among rats during the rainy season compared with rats in the dry season. There are no current available data on leptospira serovars in humans in the Mekong Delta, although existing studies suggest limited overlapping between human and rat serovars. Further studies should take into account a wider range of potential reservoirs (i.e., dogs, pigs) as well as perform geographically linked co-sampling of humans and animals to establish the main sources of leptospirosis in the region.

Peste des petits ruminants (PPR) is a contagious disease with potential for high economic impacts. Convalescent and vaccinated small ruminants develop a strong and lifelong immunity and are fully protected against re-infection. Consequently, the presence of antibodies, whether they are against a wild virus or a vaccine, is a marker of the host immune protection. Thus, specific antibodies cannot be missed with classical and validated diagnosis assays, such as conventional enzyme-linked immunosorbent assay (ELISA), an alternative testing method to the conventional virus neutralization test (VNT), allowing to monitor vaccination or locate disease outbreaks. Recent insights into the virions' protein composition, development of expression systems, recombinant vectors to express protective antigens, have contributed greatly to update our current knowledge about diagnostics. In the light of this progress, the chapter focuses on PPRV serological tests with particular attention to diagnostic antigen targets and refinement of assays for accurate serodiagnosis of PPRV.

Three hundred and ten blood samples from clinically healthy and suspected camels (male and female) of four breeds from different location of Al-jouf (Saudi Arabia), were examined to detect prevalence rate of T. evansi infection by CATT test. An overall prevalence was determined as 43.8%. The variability of trypanosomosis was highly significant according to the factor moving, clinical signs and animal status regarding it age, sex and lactation status. Prevalence rate increased significantly in May and a second peak was observed in September.