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PURPOSE: The objective of this study is to compare the diagnostic value of commercial enzyme-linked immunosorbent assay (ELISA) with indirect immunofluorescent antibody test (IFA) in detecting immunoglobulin-G (IgG) antibodies to Ehrlichia canis and Anaplasma phagocytophilum. METHODS: Seventy-four serum samples, obtained from dogs believed to be naturally infected with E. canis or A. phagocytophilum, were analyzed. RESULTS: By ELISA, 48 (64.9%) samples were found positive for IgG to E. canis, 10 (13.5%) to A. phagocytophilum, 12 (16.2%) to both E. canis and A. phagocytophilum, and in 4 (5.4%) samples no presence of antibodies was detected. The number of serologically positive dogs for IgG was 44 (59.5%) to E. canis, 10 (13.5%) to A. phagocytophilum, 16 (21.6%) to both E. canis and A. phagocytophilum, and 4 (5.4%) were determined negative by means of IFA. In most samples the antibody titer did not exceed 1:80 but in 5 it reached a level of 1:320, and in other 4 of even above 1:640. CONCLUSIONS: This study shows that IFA assay is more sensitive than commercial ELISA rapid test when serum antibody titers are low.

Human metapneumovirus (hMPV) of the family Paramyxoviridae is a relatively new virus causing severe acute respiratory tract infections (SARI) in children. Data on hMPV infection in Asia including Sri Lanka is limited. We aimed to detect respiratory viruses including hMPV in a selected group of children affected by a small outbreak of SARI presented to the Teaching Hospital, Peradeniya (THP), Sri Lanka in 2014. Nasopharyngeal aspirates (NPA) were obtained from 21 children with SARI and tested for hMPV, influenza A and B, parainfluenza 1, 2 and 3 (PIV 1–3), adenovirus and respiratory syncytial virus (RSV) antigens using an immunofluorescence assay (IFA). In addition, a one step RT-PCR was done for the detection of hMPV from the viral RNA extracts. Of the 21 NPA samples tested for respiratory viral antigens by IFA, two were positive for RSV (9.5%), one was positive for influenza A (4.8%) and one was positive for both adenovirus and PIV-2 (4.8%). Of the 21 NPA viral RNA extracts tested by RT-PCR, 18 (86%) were positive for hMPV, in which 2 were co-infected with RSV and influenza A virus, respectively. hMPV was the predominant cause of SARI outbreak (2014) in children presented to the THP, Sri Lanka.

Made available in DSpace on 2019-10-06T16:23:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01. Added 1 bitstream(s) on 2019-10-09T18:35:21Z : No. of bitstreams: 1 S1984-29612019000100180.pdf: 1026926 bytes, checksum: 3554892a2d6c8d0aed35572f3b2c37fc (MD5) Ehrlichiosis is caused by agents belonging to Ehrlichia genus. Despite the frequent reports on the serological and molecular detection of E. canis in dogs in Brazil, there is scant data on ehrlichiosis in brazilian cats. This study aimed at investigating the occurrence of Ehrlichia spp. in domestic cats from Greater Rio de Janeiro, and evaluating hematological changes associated with this rickettsial infection. We searched for IgG antibodies against E. canis on blood samples of 216 cats by Indirect Fluorescence Assay (IFA). Additionally, we performed nested PCR (nPCR) and real-time PCR (qPCR) assays targeting E. canis-16S rRNA and dsb gene, respectively. Fifty-seven (26.4%) cats were seropositive for Ehrlichia spp. by IFA. Ehrlichia spp.-16S rRNA gene fragments were detected in 3 cats (1.4%). Although the obtained 16S rRNA sequences showed 99 to 100% identity with E. canis, cats were negative in qPCR. Anemia, thrombocytopenia, leukocytosis, left shift neutrophil and hyperproteinemia were observed. Anemia was statistically associated with seropositivity to E. canis and kittens showed lower positivity rates (p

Porcine circovirus type 3 (PCV3) is a novel porcine circovirus associated with porcine dermatitis and nephritis syndrome (PDNS), reproductive failure, and multisystemic inflammation. Capsid protein (Cap) encoded by PCV3 ORF2 gene has been identified as an immunogenic protein. Currently, there is no immunofluorescence assay (IFA) available for serological diagnosis. Here, the N-terminal 33 amino acids of Cap protein were predicted to serve as a PCV3 nuclear localization signal (NLS). Two types of recombinant plasmids were constructed for recombinant protein expression in Sf9 cells by using a baculovirus expression system: plasmid rvBac-Pc for full-length Cap protein expression and rvBac-Sc for Cap protein expression with a honeybee melittin signal peptide in place of the predicted NLS sequence. Expression of the nuclear localization sequences was further analyzed by IFA. Strong and specific fluorescence signals were observed in the nucleus of rvBac-Pc-transfected cells and in the cytoplasm of rvBac-Sc-transfected cells. No cross-reactivity was observed with porcine circovirus type 2, porcine pseudorabies virus, classical swine fever virus, or porcine reproductive and respiratory syndrome virus. In summary, we developed two fluorescence detection modes for Cap protein that can be used to detect PCV3 antibodies. This method is suitable for the diagnosis and epidemiological investigation of PCV3. This study provides a reliable detection method for monitoring PCV3 antibody level in pigs in the future.

Cat-associated Bartonella species, which include B. henselae, B. koehlerae, and B. clarridgeiae, can cause mild to severe illness in humans. In the present study, we evaluated 1362 serum samples obtained from domestic cats across the U.S. for seroreactivity against three species and two strain types of Bartonella associated with cats (B. henselae type 1, B. henselae type 2, B. koehlerae, and B. clarridgeiae) using an indirect immunofluorescent assay (IFA). Overall, the seroprevalence at the cutoff titer level of ≥1:64 was 23.1%. Seroreactivity was 11.1% and 3.7% at the titer level cutoff of ≥1:128 and at the cutoff of ≥1:256, respectively. The highest observation of seroreactivity occurred in the East South-Central, South Atlantic, West North-Central, and West South-Central regions. The lowest seroreactivity was detected in the East North-Central, Middle Atlantic, Mountain, New England, and Pacific regions. We observed reactivity against all four Bartonella spp. antigens in samples from eight out of the nine U.S. geographic regions.

The genus Ehrlichia includes tick-borne bacterial pathogens affecting humans, domestic and wild mammals. Ehrlichia minasensis has been identified in different animal species and geographical locations, suggesting that this is a widely distributed and generalist Ehrlichia. In the present study, we evaluated Ehrlichial infection in 148 Equidae presented to the Medical Clinic Department of a Veterinary Hospital from a midwestern region of Brazil. Blood samples and ticks collected from the animals were tested by Polymerase Chain Reaction (PCR) for the presence of Ehrlichia spp. A multigenic approach including Anaplasmataceae-specific (i.e., 16S rRNA, groEL, gltA) and Ehrlichia-specific (i.e., dsb and trp36) genes was used for accurate bacteria identification. Sera samples were also collected and evaluated for the detection of anti-Ehrlichia antibodies by indirect fluorescent antibody test (IFA). Possible associations between molecular and serological diagnostics and clinical and hematological manifestations were tested using chi-squared or Fisher’s exact tests. Sequence analysis of the dsb fragment revealed that three horses (2.03%) were exposed to E. minasensis. Sixty-one (41.2%) Equidae (58 equines and three mules), were seropositive for Ehrlichia spp., with antibody titers ranging between 40 and 2560. Seropositivity to ehrlichial antigens was statistically associated with tick infestation, rural origin, hypoalbuminemia and hyperproteinemia (p ≤ 0.05). The present study reports the first evidence of natural infection by E. minasensis in horses from Brazil.

International audience; Ehrlichia spp. are important tick-borne pathogens of animals in Brazil, and Ehrlichia canis is the most prevalent species infecting dogs. Moreover, Ehrlichia minasensis has also recently been identified as a novel ehrlichial agent that infects cattle in Brazil. The objective of this study was to determine whether dogs could be infected by E. minasensis. To investigate this possibility, sera (n = 429) collected from dogs in the Pantanal region were retrospectively analyzed for the presence of antibodies against E. canis and E. minasensis. Canine sera were screened by two isolates of E. canis in indirect immunofluorescence assay (IFA) and the majority (n = 298; 69.4%) had antibodies with endpoint titers ranging from 80 to 327,680. In order to further confirm E. canis-specific antibodies, IFA positive sera were analyzed by ELISA using E. canis-specific peptides (i.e. TRP19 and TRP36 US/BR/CR), which detected E. canis antibodies in 80.2% (239/298) of the dog sera. Fifty-nine (13.7%) samples had detectable antibodies to E. canis by IFA but were negative by E. canis peptide ELISA. These sera were then tested by E. minasensis IFA (Cuiaba strain) as antigen and 67.8% (40/59) were positive (titers ranging from 80 to 20,480). Eleven sera had antibody titers against E. minasensis at least two-fold higher than observed for E. canis and suggests that these dogs were previously infected with E. minasensis. The results of the present study suggest that multiple ehrlichial agents infect dogs in Brazil, which highlights the need to consider different Ehrlichia spp. in Brazilian dogs, particularly in areas where dogs are frequently exposed to multiple tick species. This investigation is the first to provide serologic evidence of E. minasensis infection in dogs from Brazil.

Borrelia burgdorferi is a bacterial pathogen transmitted by Ixodes ticks and is responsible for Lyme disease in both humans and dogs. The aim of this work was to evaluate B. burgdorferi diffusion among stray dogs in Palermo (Sicily, Italy) by serological methods in order to study the risk factors associated with the infection. Serum and blood samples of 316 dogs were collected from a shelter in Palermo, and were analyzed for the presence of antibodies against B. burgdorferi by indirect immunofluorescence assay (IFA), and of the ospA gene by real-time PCR, respectively. Seventeen sera (5.4%) were positive for the antibodies via IFA and one blood (0.3%) for ospA via real time PCR. On the basis of serological results, the evaluation of the potential risk factors (sex, age, breed and coat color) was carried out. The multivariate analysis indicated that male sex is a factor significantly associated with B. burgdorferi seropositivity. This study confirms that male dogs have a higher risk of developing the disease than females, and represents the first investigation on the spread of B. burgdorferi among stray dogs in Sicily.

A total of 100 domestic cats from Luanda (Angola) were tested for the presence of antibodies against Bartonella henselae and spotted fever group of Rickettsia (SFGR) using indirect immunofluorescence assay (IFA). Molecular screening targeting the riboflavin synthase (ribC) gene for Bartonella and outer membrane protein B (ompB) gene for Rickettsia, using conventional PCR and sequencing was also performed in cat´s blood samples. Sixty-six percent of the cats from Luanda had IgG antibodies against Bartonella species but none of them had antibodies against SFGR. Of the total seroreactive cats for Bartonella henselae, 4.5% had an IgG titre of 64 (cut-off), 60.6% a titre of 128, 28.8% a titre of 256 and 6.1% a titre of 512. A statistically significant association was observed between seropositivity for Bartonella henselae and the lack of access to prophylaxis against ectoparasites (p = 0.018). Molecular detection and further sequence analysis of the positive amplicons allowed identification of Bartonella henselae in a 2-year old male cat. To the best of our knowledge this study confirms for the first time, the presence of Bartonela henselae circulating in domestic cats from Luanda. This fact call the attention for the possible cases of cat-scratch disease in humans. P. Barradas (SFRH/BD/116449/2016) acknowledges FCT, the Portuguese Foundation for Science and Technology for financial support. IPATIMUP integrates the i3S Research Unit, which is partially supported by FEDERthrough the Operational Programme for Competitiveness Factors-COMPETE and National Funds through FCT, under the project number PEst-C/SAU/LA0003/2013. This paper was published under the framework of the European Social Fund, Human Resources Development Operational Programme 2007–2013 (POSDRU/159/1.5/S/136893). info:eu-repo/semantics/publishedVersion

In the indeterminate chronic period of Chagas disease (ChD) the treatment has not been conclusive, because the serological negativization requires many years. This study aims to evaluate the efficacy of nifurtimox (NF) in the treatment of chronic ChD in prolonged follow-up by serological techniques of indirect immunofluorescence assay (IFA) and enzyme-linked immunosorbent assay (ELISA) IgG comparing 2 groups of patients, treated and non treated. Mann-Whitney test was performed for ELISA and IFA, with significant difference between the groups (Plt; 0.05). IgG levels were lower in individuals treated compared with untreated patients, indicating chemotherapeutic efficacy in prolonged follow-up.

Made available in DSpace on 2020-12-12T02:14:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01. Added 1 bitstream(s) on 2021-07-15T15:25:56Z : No. of bitstreams: 1 S1984-29612020000200407.pdf: 187601 bytes, checksum: 9bd904f85dcfa2daf768578ff21cf013 (MD5) We evaluated the prevalence of anti-Toxoplasma gondii antibodies in the serum samples collected from domestic cats in Belém, Pará, Brazil. We also correlated the presence of T. gondii antibodies with environmental variables and cat-owner habits. Four-hundred and forty-seven serum samples from domestic cats were analyzed. The sera were tested using an indirect immunofluorescence assay. Among the animals analyzed, 21.92% (98/447) were seropositive. A statistically significant association was found in relation to age and serology among the animals over 1 year old (p

The purpose of this study was to do a serological survey on three rickettsial species: Rickettsia rickettsii and Rickettsia parkeri, two species of the spotted fever group (SFG) that are considered to be great importance for public health; and Rickettsia bellii, a species of unknown pathogenicity that infects a variety of human-biting ticks. Serum samples from 273 dogs were tested using the indirect immunofluorescence assay (IFA). A total of 52 samples (19.04%) were seropositive for at least one of the three Rickettsia spp. antigens. Thirty-eight (73.07%), twelve (23.07%) and one (1.92%) of these dogs showed homologous reactions to R. bellii, R. rickettsii and R. parkeri, respectively. Our results showed that the seroprevalence of Rickettsia spp. was relatively low. However, the positive serological tests indicated that these dogs had become infected by these agents at some point in their lives. Lastly, our study adds to the previous knowledge on the epidemiology of rickettsiosis in the state of Goiás by doing the first record of detection of anti-R. rickettsii, R. parkeri and R. bellii antibodies by IFA among dogs, thus indicating that these agents may be circulating in the dog population analyzed. Resumo O objetivo deste estudo foi realizar um levantamento sorológico para três espécies de rickettsias: Rickettsia rickettsii e Rickettsia parkeri, duas espécies do grupo da febre maculosa (GFM) consideradas de grande importância para a saúde pública; e Rickettsia bellii, uma espécie de patogenicidade desconhecida que infecta uma variedade de carrapatos que parasitam seres humanos. Amostras de soro de 273 cães foram testadas, usando-se a técnica de reação de imunofluorescência indireta (RIFI). O total de 52 amostras (19,04%) foram soropositivas para pelo menos um dos três antígenos de Rickettsia spp. Trinta e oito (73,07%), doze (23,07%) e um (1,92%) desses cães apresentaram reações homólogas à R. bellii, R. rickettsii e R. parkeri, respectivamente. Esses resultados demonstraram uma baixa soroprevalência para Rickettsia spp. No entanto, as amostras positivas indicam que esses cães foram infectados por esses agentes em algum momento de suas vidas. Por fim, este estudo contribui para o conhecimento sobre a epidemiologia das rickettsioses no estado de Goiás, realizando a primeira detecção de anticorpos anti-Rickettsia rickettsii, R. parkeri e R. bellii pela RIFI em cães, indicando que esses agentes podem estar circulando na população canina analisada.

Tis study evaluated the presence of Neospora caninum and Toxoplasma gondii antibodies in 320 domestic dogs living in urban and rural areas of the municipality of Poconé, located in the Pantanal wetlands of the state of Mato Grosso, Brazil. Anti-N. caninum (cut-of point 50) and anti-T. gondii (cut-of point 16) antibodies were detected by means of immunofuorescence assays (IFA), using NC-Bahia and RH strains as antigens, respectively. Anti-N. caninum antibodies were detected in 69 (21.56%; 95% CI: 17.27%-26.56%) dogs, 31 (44.93%) of which lived in urban areas and 38 (55.07%) in rural areas, and endpoint titers ranged from 50 to 3200. Anti-T. gondii antibodies were found in 132 (41.25%; 95% CI: 35.84%-46.87%) dogs, 58 (43.94%) from urban areas and 74 (56.06%) from rural areas, and endpoint titers ranged from 16 to 8192. A total of 33 dogs (10.3%) (12 urban and 21 rural animals) reacted to both agents (P

We revisit a singular value decomposition (SVD) algorithm given in Chen et al. (2019b) for exploratory Item Factor Analysis (IFA). This algorithm estimates a multidimensional IFA model by SVD and was used to obtain a starting point for joint maximum likelihood estimation in Chen et al. (2019b). Thanks to the analytic and computational properties of SVD, this algorithm guarantees a unique solution and has computational advantage over other exploratory IFA methods. Its computational advantage becomes significant when the numbers of respondents, items, and factors are all large. This algorithm can be viewed as a generalization of principal component analysis (PCA) to binary data. In this note, we provide the statistical underpinning of the algorithm. In particular, we show its statistical consistency under the same double asymptotic setting as in Chen et al. (2019b). We also demonstrate how this algorithm provides a scree plot for investigating the number of factors and provide its asymptotic theory. Further extensions of the algorithm are discussed. Finally, simulation studies suggest that the algorithm has good finite sample performance., 43 pages, 5 figures

Using a large, passive, febrile surveillance program in Iquitos, Peru, we retrospectively tested human blood specimens for scrub typhus group orientiae by ELISA, immunofluorescence assay, and PCR. Of 1,124 participants, 60 (5.3%) were seropositive, and 1 showed evidence of recent active infection. Our serologic data indicate that scrub typhus is present in the Peruvian Amazon.

Made available in DSpace on 2019-10-06T16:10:50Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-02-01 Toxoplasmosis is a zoonosis present worldwide. Its protozoal aethiological agent, Toxoplasma gondii, has the ability to infect several homeothermic animals and mainly human beings. The consumption of raw or undercooked meat products containing T. gondii cysts, consumption of vegetables without washing and using non-treated water are risk factors associated to the occurrence of human toxoplasmosis. Furthermore, raw or undercooked pork is an important infection source of T. gondii to humans. Due to the importance of toxoplamosis in public health, this study focused on establish the prevalence of the disease in non-technified swine herds in the northeastern region of the State of São Paulo, Brazil, using Modified Agglutination Test (MAT) and the Indirect Immunofluorescence Assay (IFA) and the risk factors for its occurrence. In addition, the agreement among both diagnostic tests was evaluated. A low prevalence of toxoplasmosis was found at animal level (7.02%). The Fisher's exact test detected correlation between positive cases with the presence of food garden in the farm (p = 0.01) and the use of non-treated water to irrigate the food garden (p = 0.005). The agreement among tests was considered moderate (Kappa index = 0.5). The results show that toxoplasmosis is a risk for humans who consume under cooked pork meat in this region and an acceptable agreement between MAT and IFA tests. Departamento de Higiene Veterinária e Saúde Pública Faculdade de Medicina Veterinária e Zootecnia (FMVZ) Universidade Estadual Paulista (Unesp), Câmpus de Botucatu, Distrito de Rubião Junior s/n Faculdade de Ciências Agrárias e Veterinárias (FCAV) Universidade Estadual Paulista – UNESP, Câmpus de Jaboticabal, Via de Acesso Prof. Paulo Donato Castellane s/n Departamento de Higiene Veterinária e Saúde Pública Faculdade de Medicina Veterinária e Zootecnia (FMVZ) Universidade Estadual Paulista (Unesp), Câmpus de Botucatu, Distrito de Rubião Junior s/n Faculdade de Ciências Agrárias e Veterinárias (FCAV) Universidade Estadual Paulista – UNESP, Câmpus de Jaboticabal, Via de Acesso Prof. Paulo Donato Castellane s/n

Background TORCH including the pathogens of Toxoplasma gondii (TOX), rubella virus (RV), cytomegalovirus (CMV), and herpes simplex virus (HSV) causes intrauterine infections and poses a worldwide threat to women especially in pregnancy. In this study, we described the seasonal difference in TORCH infection and analyzed the anti‐TORCH IgM multipositive serum samples by the indirect immunofluorescence assays (IFA). Methods To observe the seasonal influence of the anti‐TORCH IgG and IgM antibodies, a retrospective study was conducted with 10 669 women (20–40 y old) before pregnancy from August 2016 to July 2017. Totally 199 ELISA anti‐TORCH IgM multipositive serum samples were further tested by IFAs for false‐positive analysis. Results The prevalence of positive HSV1‐IgM, RV‐IgM, HSV2‐IgM, CMV‐IgM, and TOX‐IgM in the present population was 6.30%, 2.55%, 1.94%, 1.24%, and 0.67%, respectively. Additionally, the prevalence of positive RV‐IgM, CMV‐IgM, and HSV1‐IgM was statistically different among four seasons, with the highest positive rates of RV‐IgM (4.12%) in autumn, CMV‐IgM (1.75%) in summer, and HSV1‐IgM (7.53%) in winter. The confirmatory IFAs showed that the positive rates of RUV‐IgM, CMV‐IgM, and HSV2‐IgM were significantly different from those in ELISA screening experiments. Interestingly, only 32.7% (65/199) of the TORCH IgM multipositive results were consistent with those by the IFA, indicating that cross‐reaction caused false positives were common in ELISA IgM antibody screening. Conclusion The TORCH infection displayed different prevalence among four seasons in our 12‐month retrospective study. The IgM multipositives by ELISA screening may need further confirmation analysis due to its relatively high cross‐reaction rate.

The occurrence of the zoonotic protozoan parasite Toxoplasma gondii in marine mammals remains a poorly understood phenomenon. In this study, samples from 589 marine mammal species and 34 European otters (Lutra lutra), stranded on the coasts of Scotland, Belgium, France, The Netherlands and Germany, were tested for the presence of T. gondii. Brain samples were analysed by polymerase chain reaction (PCR) for detection of parasite DNA. Blood and muscle fluid samples were tested for specific antibodies using a modified agglutination test (MAT), a commercial multi-species enzyme-linked immunosorbent assay (ELISA) and an immunofluorescence assay (IFA). Out of 193 animals tested by PCR, only two harbour porpoise (Phocoena phocoena) cerebrum samples, obtained from animals stranded on the Dutch coast, tested positive. The serological results showed a wide variation depending on the test used. Using a cut-off value of 1/40 dilution in MAT, 141 out of 292 animals (41%) were positive. Using IFA, 30 out of 244 tested samples (12%) were positive at a 1/50 dilution. The commercial ELISA yielded 7% positives with a cut-off of the sample-to-positive (S/P) ratio ≥ 50; and 12% when the cut-off was set at S/P ratio ≥ 20. The high number of positives in MAT may be an overestimation due to the high degree of haemolysis of the samples and/or the presence of lipids. The ELISA results could be an underestimation due to the use of a multispecies conjugate. Our results confirm the presence of T. gondii in marine mammals in The Netherlands and show exposure to the parasite in both the North Sea and the Eastern Atlantic Ocean. We also highlight the limitations of the tests used to diagnose T. gondii in stranded marine mammals.

SUMMARYFrom 2007 to 2010, The Netherlands experienced a major Q fever outbreak with more than 4000 notifications. Previous studies suggested that Q fever patients could suffer long-term post-infection health impairments, especially fatigue. Our objective was to assess the Coxiella burnetii antibody prevalence and health status including fatigue, and assess their interrelationship in Herpen, a high-incidence village, 7 years after the outbreak began. In 2014, we invited all 2161 adult inhabitants for a questionnaire and a C. burnetii indirect fluorescence antibody assay (IFA). The health status was measured with the Nijmegen Clinical Screening Instrument (NCSI), consisting of eight subdomains including fatigue. Of the 70·1% (1517/2161) participants, 33·8% (513/1517) were IFA positive. Of 147 participants who were IFA positive in 2007, 25 (17%) seroreverted and were now IFA negative. Not positive IFA status, but age C. burnetii antibodies.

In this study the seroprevalence of anti-Borrelia burgdorferi IgG antibodies in populations of naturally exposed dogs in the Belgrade area was determined. Serum samples of 215 dogs were tested with in-house made indirect immunofluorescent-antibody assay (IFA), enzyme-linked immunosorbent assay (ELISA), and Western blot assay (WB). Sera were from 34 pet dogs suspected of having Lyme borreliosis, 41 hunting dogs, 125 shelter dogs, and 15 clinically healthy pet dogs. Results of WB were used as a standard against which the performances of ELISA and IFA were evaluated. ELISA was significantly more sensitive than IFA (84.8 vs 67.9%), whereas the specificities of these tests were just slightly different (94.4% vs 92.6 %). Borrelia burgdorferi seroprevalence of 24.7% among tested dogs was recorded by WB test. The highest seroprevalence of 31.2% was found in the group of shelter dogs, while hunting and pet dogs showed lower seroprevalence of 19.5% and 17.6%, but the difference was statistically insignificant. These results are the first report on the seroprevalence of IgG antibodies to B. burgdorferi in dogs in the Belgrade area and could be of importance in the assessment of the risk of infection with B. burgdorferi in human population.

Although goat dairy farms in Brazil may have a higher risk of infection by Neospora caninum than beef farms, risk factor evaluation on a representative population remains to be fully established in Brazil. Accordingly, this study aimed to establish the occurrence of anti-N. caninum antibodies and factors associated with exposure in 406 blood samples from five dairy and three beef goat farms in the state of Paraíba, northeastern Brazil. Anti-N. caninum antibodies were detected by indirect immunofluorescence assay (IFA), with samples considered positive when reacting with dilution ≥ 1:50. A total of 106/406 goats (26.11%; 95% CI: 21.96-30.72%) were seroreactive comprising 2/61 (3.28%), 10/45 (22.22%), 13/50 (26.00%), 17/51 (33.33%) to 29/46 (63.04%) in dairy farms, and from 3/54 (5.56%), 12/50 (24.00%) to 20/49 (40.82%) on the beef farms. No significant associations were found in relation to age, gender, dairy versus beef farms, occurrence of abortions or mummified fetuses, and seroreactivity to N. caninum (P>0.05). In conclusion, goat farms in the state of Paraíba showed the highest occurrence of anti-N. caninum antibodies to date in Brazil. RESUMO Embora as criações caprinas de leite no Brasil possam ter maior probabilidade de risco de infecção por Neospora caninum do que as de carne, a avaliação dos fatores de risco em uma população representativa ainda não está totalmente estabelecida no Brasil. Dessa forma, este estudo teve por objetivo estabelecer a soroprevalência de N. caninum e seus fatores associados à exposição em 406 amostras de sangue de cinco fazendas de leite e três de corte provenientes do estado da Paraíba, região Nordeste do Brasil. A detecção de anticorpos anti-N. caninum foi realizada utilizando-se a reação de imunofluorescência indireta (RIFI), com as amostras consideradas positivas na diluição ≥ 1:50. No total, 106/406 (26,11%; IC 95%: 21,96-30,72%) caprinos foram sororreagentes, variando de 2/61 (3,28%), 10/45 (22,22%), 13/50 (26,00%), 17/51 (33,33%) a 29/46 (63,04%) em fazendas de leite, e de 3/54 (5,56%), 12/50 (24,00%) a 20/49 (40,82%) em fazendas de corte. Não foram observadas associações significativas entre idade, sexo, criação de leite e carne, ocorrência de abortamentos ou fetos mumificados e sororreatividade para N. caninum (P>0,05). Em conclusão, fazendas de caprinos da Paraíba mostraram as mais altas ocorrências de anticorpos anti-N. caninum até o momento no Brasil.

Graphical abstract, Highlights • Giardia intestinalis (48/177) and Cryptosporidium parvum (98/177) occur in Austrian calves. • C. parvum genotypes IIaA15G2R1, IIaA19G2R2, IIaA21G2R1 and IIaA14G1R1 were found. • Giardia intestinalis assemblage E and a single sample of assemblage A were found. • Cryptosporidium was detected mostly in the first two weeks of age (72/102). • Point-of-care test validity varied greatly for Giardia and Cryptosporidium., To obtain information about the occurrence and genotype distribution of G. intestinalis and C. parvum in Austrian cattle, faecal samples from diarrhoeic calves younger than 180 days of age originating from 70 farms were examined. Of the 177 faecal samples, 27.1% were positive for Giardia cysts (immunofluorescence microscopy) and 55.4% for Cryptosporidium oocysts (phase-contrast microscopy). Positive samples were characterized by nested PCR for Giardia, 83.3% (triosephosphate isomerase; tpi) and 89.6% (β-giardin; bg) were positive, while the Cryptosporidium nested PCR returned 92.5% (60-kDa glycoprotein) positive results. Sequence analysis revealed one assemblage A-positive sample and 30 (bg) respectively 29 (tpi) assemblage E-positive samples for G. intestinalis. For C. parvum four subtypes within the IIa family (IIaA15G2R1, n = 29; IIaA19G2R2, n = 3; IIaA21G2R1, n = 2; IIaA14G1R1, n = 1) could be differentiated. Validation of two immunochromatographic point-of-care tests resulted in a sensitivity of 29.2% and 77.6%; a specificity of 98.4% and 91.1% for the detection of Giardia intestinalis and Cryptosporidium parvum, respectively. Results confirm the widespread occurrence of both protozoa in diarrhoeic calves in Austria.

SUMMARY The aims of this study were to determine the seroprevalence of Ehrlichia spp. and risk factors for exposure in a restricted population of dogs, horses, and humans highly exposed to tick bites in a Brazilian rural settlement using a commercial ELISA rapid test and two indirect immunofluorescent assays (IFA) with E. canis and E. chaffeensis crude antigens. Serum samples from 132 dogs, 16 horses and 100 humans were used. Fifty-six out of 132 (42.4%) dogs were seropositive for E. canis. Dogs > one year were more likely to be seropositive for E. canis than dogs ≤ one year (p = 0.0051). Ten/16 (62.5%) and 8/16 (50%) horses were seropositive by the commercial ELISA and IFA, respectively. Five out of 100 (5%) humans were seropositive for E. canis and E. chaffeensis. Rhipicephalus sanguineus (n = 291, 97.98%) on dogs and Amblyomma cajennense (n = 25, 96.15%) on horses were the most common ticks found. In conclusion, anti-Ehrlichia spp. antibodies were found in horses; however, the lack of a molecular characterization precludes any conclusion regarding the agent involved. Additionally, the higher seroprevalence of E. canis in dogs and the evidence of anti-Ehrlichia spp. antibodies in humans suggest that human cases of ehrlichiosis in Brazil might be caused by E. canis, or other closely related species. RESUMO Objetivou-se determinar a soroprevalência de Ehrlichia spp. e os fatores de risco associados a exposição em uma população restrita de cães, cavalos e humanos altamente expostos a picadas de carrapatos em um assentamento rural brasileiro utilizando um teste comercial de ELISA rápido e dois testes de imunofluorescência indireta (IFI) com antígenos brutos de E. canis e E. chaffeensis. Amostras de soro de 132 cães, 16 cavalos e 100 humanos foram utilizadas. Cinquenta e seis/132 (42,4%) cães foram soropositivos para E. canis. Cães > um ano apresentaram mais chance de serem soropositivos para E. canis do que cães ≤ um ano (p = 0,0051). Dez/16 (62,5%) e 8/16 (50%) cavalos foram soropositivos pelo ELISA comercial e IFI, respectivamente. Cinco/100 (5%) humanos foram soropositivos para E. canis e E. chaffeensis. Rhipicephalus sanguineus (n = 291, 97,98%) nos cães e A. cajennense (n = 25, 96,15%) nos cavalos foram os carrapatos mais encontrados. Concluindo, anticorpos anti-Ehrlichia spp. foram encontrados em cavalos; entretanto, a ausência de uma caracterização molecular impede qualquer conclusão sobre agente envolvido. Além disso, a alta soroprevalência de E. canis em cães e a evidência de anticorpos anti-Ehrlichia sp. em humanos, sugere que os casos de erliquiose humana no Brasil possam ser causados por E. canis ou outra espécie intimamente relacionada.

BACKGROUND: At least two genetically different porcine epidemic diarrhea virus (PEDV) strains have been identified in the United States (U.S. PEDV prototype and S-INDEL-variant strains). The current serological assays offered at veterinary diagnostic laboratories for detection of PEDV-specific antibody are based on the U.S. PEDV prototype strain. The objectives of this study were: 1) isolate the U.S. PEDV S-INDEL-variant strain in cell culture; 2) generate antisera against the U.S. PEDV prototype and S-INDEL-variant strains by experimentally infecting weaned pigs; 3) determine if the various PEDV serological assays could detect antibodies against the U.S. PEDV S-INDEL-variant strain and vice versa.RESULTS: A U.S. PEDV S-INDEL-variant strain was isolated in cell culture in this study. Three groups of PEDV-negative, 3-week-old pigs (five pigs per group) were inoculated orally with a U.S. PEDV prototype isolate (previously isolated in our lab), an S-INDEL-variant isolate or virus-negative culture medium. Serum samples collected at 0, 7, 14, 21 and 28 days post inoculation were evaluated by the following PEDV serological assays: 1) indirect fluorescent antibody (IFA) assays using the prototype and S-INDEL-variant strains as indicator viruses; 2) virus neutralization (VN) tests against the prototype and S-INDEL-variant viruses; 3) PEDV prototype strain whole virus based ELISA; 4) PEDV prototype strain S1-based ELISA; and 5) PEDV S-INDEL-variant strain S1-based ELISA. The positive antisera against the prototype strain reacted to and neutralized both prototype and S-INDEL-variant viruses, and the positive antisera against the S-INDEL-variant strain also reacted to and neutralized both prototype and S-INDEL-variant viruses, as examined by IFA antibody assays and VN tests. Antibodies against the two PEDV strains could be detected by all three ELISAs although detection rates varied to some degree.CONCLUSIONS: These data indicate that the antibodies against U.S. PEDV prototype and S-INDEL-variant strains cross-reacted and cross-neutralized both strains in vitro. The current serological assays based on U.S. PEDV prototype strain can detect antibodies against both U.S. PEDV strains.

Toxoplasmosis in sheep is a disease of great importance in veterinary medicine, which causes economic losses in livestock and has a great impact on human health, since consumption of infected meat facilitates transmission of zoonotic infections. Blood samples from sheep (n = 360) were collected from 13 farm properties in the municipality of Lages, Santa Catarina, to estimate the prevalence of toxoplasmosis and identify risk factors associated with Toxoplasma gondii infection. T. gondii, antibodies were investigated by means of the indirect immunofluorescence assay (IFA) and enzyme-linked immunosorbent assay (ELISA). Animals infected with T. gondii were found on 100% of the farms. IFA detected 56.9% (205/360) and ELISA 42.5% of the infected sheep. Breed was the only risk factor associated with the presence of T. gondii antibodies. ELISA showed sensitivity of 61%, specificity of 82% and kappa of 0.41, which was considered moderate. This allows use of ELISA as an alternative technique for diagnosing T. gondii in sheep. A toxoplasmose ovina é uma doença parasitária de elevada importância em medicina veterinária e em saúde pública, acarretando prejuízos na produção animal, gerados pelas perdas reprodutivas e econômicas, além de sua implicação na saúde humana, já que o consumo de carne infectada facilita a transmissão zoonótica. Para determinar a prevalência e identificar fatores de risco para a infecção por T. gondii em ovinos de Lages, Santa Catarina, amostras de sangue (n = 360) foram coletadas em 13 propriedades. Cada criador respondeu a um questionário para permitir a identificação dos fatores de risco da infecção. A pesquisa de anticorpos foi realizada por meio da Reação de Imunofluorescência Indireta (RIFI > 64) e do Ensaio Imunoenzimático Indireto (ELISA). Em 100% das propriedades foram encontrados animais positivos. Pela RIFI, 205 (56,94%) ovinos apresentaram anticorpos contra T. gondii e pelo ELISA, 153 (42,50%). Considerando-se as técnicas sorológicas e a análise estatística, foram fatores de risco pelo ELISA: a idade, a fonte de água e a categoria animal; e pela RIFI, o tipo racial. Foi constatada sensibilidade de 61%, especificidade de 82% e concordância Kappa de 0,41 entre o ELISA e a RIFI (1:64), considerada moderada, permitindo indicar o ELISA como técnica adequada para o diagnóstico de T. gondii na espécie ovina.

Until now there is no evidence that chlamydiosis does exist among cats in the Republic of Croatia regardless of the fact that feline chlamydiosis is a worldwide disease. This report describes the clinical examination of a one year old cat with bilateral conjunctivitis, as well as the diagnostic methods used to confirm infection with Chlamydophila (Cp.) felis. We have used rapid enzyme immunoassay test (EIA) for antigen detection, in order to examine the swabs taken from the eyes and nostrils. The positive result was confirmed by direct immunofluorescence (DIF) made on scrapings from cat's conjunctivas. Conventional polymerase chain reaction (PCR) made on swabs taken from cat's conjunctivas and nostrils and sequencing the PCR product was used to confirmed infection of cat precisely with Chlamydophila felis. No increase of IgG antibodies against chlamydias was noted using indirect immunofluorescence (IFA) method.

Made available in DSpace on 2022-04-28T20:43:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-01-01 Toxoplasmosis is a worldwide zoonosis caused by Toxoplasma gondii that can infect a large variety of animals, including humans. The present study aimed to evaluate the frequency of auti-T.gondii antibodies in dogs from a peripheral district of Botucatu and to establish the association among some epidemiological variables in order to evaluate risk factors for toxoplasmosis infection. Serum samples from dogs were screened using an indirect fluorescent antibody (IFA) test. Anti-T.gondii antibody prevalence was 56%. The highest titer was 1024 (1.79%) and the most frequent titers were 16 (57.14%) and 64 (33.93%). The chi-square (X2) test revealed significant association among variables such as dog access to street, ingestion of raw meat and presence of synantropic animals in the domestic environment. These results demonstrate that toxoplasmosis is present in dogs from Jardim Santa Elisa district. Department of Veterinary Hygiene and Public Health Veterinary Medicine and Animal Husbandry School São Paulo State University UNESP, Botucatu, São Paulo State Department of Pediatrics Botucatu Medical School São Paulo State University UNESP, Botucatu, São Paulo State Departamento de Higiene Veterinária e Saúde Pública Faculdade de Medicina Veterinária e Zootecnia Universidade Estadual Paulista UNESP, Distrito de Rubião Junior, s/n, 18618-000, Botucatu, SP Department of Veterinary Hygiene and Public Health Veterinary Medicine and Animal Husbandry School São Paulo State University UNESP, Botucatu, São Paulo State Department of Pediatrics Botucatu Medical School São Paulo State University UNESP, Botucatu, São Paulo State Departamento de Higiene Veterinária e Saúde Pública Faculdade de Medicina Veterinária e Zootecnia Universidade Estadual Paulista UNESP, Distrito de Rubião Junior, s/n, 18618-000, Botucatu, SP

This chapter presents the most commonly used serological methods for the diagnosis of Chikungunya virus (CHIKV) infection in humans. CHIKV is a mosquito-borne Alphavirus widely distributed in the tropical and subtropical regions of Africa, Asia, and America. CHIKV infection in human causes acute febrile illness frequently accompanied by severe joint pain. Most of the infected patients may develop chronic arthralgia that may persist for several months or years. Laboratory diagnosis of CHIKV infection is mainly based on molecular and serological tests. The serological tests represent a valuable tool for diagnosis and epidemiological studies. Enzyme-linked immunosorbent assay (ELISA) and immunofluorescence assay (IFA) are simple, rapid, and sensitive techniques widely used for the diagnosis of CHIKV infection. However, these methods represent a screening tool and often require confirmation by a second-line assays. Serum virus neutralization assay is more specific than ELISA and IFA tests and is considered a confirmatory test. Neutralization assay is employed to determine the titer of virus neutralizing antibodies against CHIKV in patients' sera. The basis of microneutralization assay (MNA), results interpretation, and procedures will be illustrated in this chapter.

The sensitivity of a K39 ELISA (Leishmania IgG, Virion/Serion) for the detection of antibodies in patients with imported leishmaniasis was compared with an immunofluorescence assay (IFA), which was applied as “golden standard”. The retrospective study comprised 93 IFA-positive or borderline sera from 42 patients with visceral (n = 16) or cutaneous (n = 26) leishmaniasis. Patients had acquired infection predominately in the Mediterranean area or the Middle East. The Leishmania species (Leishmania donovani/infantum, Leishmania tropica, Leishmania major) were identified by real-time PCR. The majority (94 %) of first samples from patients with visceral leishmaniasis (VL) tested positive by K39 ELISA. Antibody levels ranged from low to very high (33.19–1990.00 U/ml; median 596.66 U/ml) but did not correlate with the respective IFA titers. High K39 ELISA values correlated with acute infection in immunocompetent individuals. K39 antibodies declined in all individuals after clinically successful therapy, but time to seronegativity varied considerably (51 weeks to >6 years). In patients with cutaneous leishmaniasis (CL), the sensitivity of the K39 ELISA was low (23 %) compared to IFA (92 % positive). Antibody levels ranged from low to medium (10.85–524.77 U/ml; median 19.77 U/ml). The highest antibody concentrations were seen in L. infantum-infected individuals. Summarizing, a high K39 ELISA value indicates active VL. The assay is, like IFA, not a measure for effective therapy but may support post-treatment monitoring. Low level positivity can indicate subclinical, previous or clinically cured VL or even CL. The K39 ELISA can supplement highly sensitive screening tests in the diagnosis and follow-up of imported leishmaniasis.

An enzyme linked immunosorbent assay (ELISA) was used to evaluate antibody positive titer in vaccinated and non-vaccinated cattle using schizont infected myeloid cells as an antigen. The result was compared with indirect fluorescent antibody level in the same animals. For this study 116 milking cows, 95 vaccinated and 21 non-vaccinated, were bleeded in order to prepare sera. They were tested with both ELISA and IFA tests. 94 sera had positive antibody titer and 22 sera were negative through ELISA test but, with IFA test, only 89 sera showed positive antibody titer and 27 were negative. Thereby, it was concluded that the sensitivity and specificity of ELISA test in comparison with IFA test was 95.5 % and 66.6 % respectively. This study generally indicated that ELISA could be an effective test for seroepidemiological investigations of bovine tropical theileriosis, and it is considered to be valid as an additional test to distinguish the vaccinated from the non vaccinated cattle in order to schedule vaccination programs.

From a cross-sectional observational study with convenience samples, 347 blood samples from horses were collected from different physiographic regions, as follows: Santa Catarina Plateau (Santa Catarina State - SC), Médio Paraíba do Sul (São Paulo State - SP and Rio de Janeiro State RJ), Mountainous and Metropolitan regions (Rio de Janeiro State - RJ). Samples were tested for the presence of antibodies (IgG) anti Neorickettsia risticii by indirect immunofluorescence assay (IFA). The frequency obtained in this study corroborates with the ones obtained in the U.S.A., which refers to endemic regions. Fisher's exact test showed significant differences in the number of positive animals between regions, indicating that the probability of an animal becoming infected varies depending on the area. The CI 95% revealed no association between infection and geopolitical space. Moreover, Odds ratio test showed differences of an animal getting infected in different regions. This event could be influenced by the type of treatment used in each area, as the seasonal frequency of injury or even potential vectors. Therefore, there are seropositive animals for N. risticii in the studied areas, suggesting that this agent may be circulating in those regions. Future studies mainly based on molecular analyzes are needed to confirm these serological findings. A partir de um delineamento observacional transversal com amostras de conveniência, 347 amostras de sangue foram coletadas de diferentes regiões fisiográficas: Planalto de Santa Catarina (Estado de Santa Catarina - SC), Região do Médio Paraíba do Sul (Estados de São Paulo - SP e Rio de Janeiro - RJ), Região Serrana e Metropolitana (ambas do Estado do Rio de Janeiro - RJ). As amostras foram testadas para a presença de anticorpos (IgG) anti-Neorickettsia risticii por imunofluorescência indireta (IFI). A prevalência obtida no presente estudo corrobora com demais resultados obtidos nos Estados Unidos da América. O Teste Exato de Fisher demonstrou diferença significativa no número de animais positivos entre as regiões, indicando assim que a probabilidade de um animal se infectar varia dependendo da região. O intervalo de confiança (IC 95%) revelou não haver associação entre a infecção e o espaço geopolítico, este evento pode ser influenciado pelo tipo de tratamento em cada área, como sazonalidade do agravo ou frequência de potenciais vetores. Assim, a soropositividade ora encontrada sugere a circulação de N. risticii nas áreas estudadas. Estudos futuros baseados, principalmente, em análises moleculares serão importantes para a confirmação dos achados sorológicos no presente trabalho.

Ehrlichiosis is a worldwide distributed disease caused by different bacteria of the Ehrlichia genus that are transmitted by arthropod vectors. Its occurrence in dogs is considered endemic in several regions of Brazil. Regarding cats, however, few studies have been done and, consequently, there is not enough data available. In order to detect Ehrlichia spp. in cats from the central-western region of Brazil, blood and serum samples were collected from a regional population of 212 individuals originated from the cities of Cuiabá and Várzea Grande. These animals were tested by the Immunofluorescence Assay (IFA) and the Polymerase Chain Reaction (PCR) designed to amplify a 409 bp fragment of the dsb gene. The results obtained show that 88 (41.5%) cats were seropositive by IFA and 20 (9.4%) cats were positive by PCR. The partial DNA sequence obtained from PCR products yielded twenty samples that were found to match perfectly the Ehrlichia canis sequences deposited on GenBank. The natural transmission of Ehrlichia in cats has not been fully established. Furthermore, tick infestation was not observed in the evaluated cats and was not observed any association between age, gender and positivity of cats in both tests. The present study reports the first serological and molecular detection of E. canis in domestic cats located in the endemic area previously mentioned.

The general aim of this study, which was conducted for the first time in Iran, was to evaluate the seroprevalence and geographical distribution of Ehrlichia canis in a dog population in Iran, followed by molecular confirmation using PCR and sequencing. Blood samples were collected from 240 dogs in different areas of Alborz and Tehran Provinces and initially analyzed using the immunofluorescent antibody (IFA) test to detect anti-Ehrlichia canis IgG antibodies. Subsequently, nested PCR was performed based on a fragment of the 16S rRNA gene of E. canis on serologically positive samples. The results showed that 40/240 dogs (16.6%) presented anti-Ehrlichia canis IgG antibodies and that nine of the blood samples from the 40 seropositive dogs (22.5%) contained E. canis DNA, which was confirmed by sequencing. The seroprevalence of E. canis tended to be higher in purebred, one to three-year-old male dogs living in the Plain zone, in rural areas; however, this difference was not statistically significant. O objetivo geral deste estudo, que foi feito pela primeira vez no Irã, foi avaliar a soroprevalência e distribuição geográfica de Ehrlichia canis em população de cães no Irã, seguida da confirmação molecular por meio de PCR seguida de sequenciamento. Amostras de sangue de 240 cães de diferentes áreas das Províncias de Alborz e Teerã foram coletadas e, inicialmente, analisadas pelo Reação de Imunofluorescência (IFA) para detecção de anticorpos IgG anti-Ehrlichia canis Subsequentemente, uma reação do tipo nested PCR baseada em um fragmento do gene 16S rRNA de E. canis foi realizada nas amostras sorologicamente positivas. Os resultados mostraram que 40/240 cães (16,6%) apresentaram anticorpos IgG anti- Ehrlichia canis e nove (22,5%) das amostras de sangue dos 40 cães soropositivos continham DNA de E. canis, confirmado por sequenciamento. A soroprevalência de E. canis, embora não estatisticamente significativa, mostrou uma tendência em se apresentar maior em cães machos com 1-3 anos, de raça pura, que vivem em zonas planas e áreas rurais.

To the Editor: Recently, experts identified Q fever, caused by the small, gram-negative bacterium Coxiella burnetii, as an important underdiagnosed childhood disease (1). Studies on Q fever in children 1:64 for phase II immunoglobulin (Ig) G and >1:24 for phase II IgM with sensitivity (specificity) of 97.2% (100%) and 100% (56.3%), respectively. IgM testing was only performed on IgG-positive children. Positive and negative controls were run on each IFA slide. Relative risks (RR) for characteristics of children were calculated by χ2 test; p64 (Figure, panels C and D). On the day of the serosurvey 71 (32.4%) of 219 children had fever (measured body temperature >38°C or reported fever within the previous 48 hours). Test outcome did not appear to be influenced by P. falciparum infection, since 4 of 37 IgG-positive children (23 of 182 IgG-negative children) had clinical malaria, 11/37 (62/182) had asymptomatic parasitemia, and 6/37 (38/182) had fever without parasitemia, and there were no significant differences between groups. The frequency of prior malaria episodes also did not influence antibody response. Three aparasitemic children had positive phase II IgM titers (24, 96, and 1,536; phase II IgG 64, 64, and 4,096, respectively). The child with the high IgM and IgG titers was clinically ill with nonsevere C. burnetii pneumonia. This child was among 10 (27%) of 37 phase II IgG-positive children with detectable anti–C. burnetii phase I antibodies. Of all sociodemographic characteristics under consideration, only maternal illiteracy was associated with positive phase II IgG testing (RR 2.1, 95% confidence interval 1.0–4.2, p64 for C. burnetii phase II IgG was applied; B) ...

International audience; Objectives: The main objective of this study was to assess the prevalence of IgG antibodies against Toscana virus (TOSV) by an ELISA test and to determine the extent of its circulation in Tunisia. Methods: An indirect ELISA test was performed to detect anti-TOSV IgG. The results were compared to those of an indirect fluorescent antibody (IFA) test. Results: The survey tested 494 healthy people from various regions of Tunisia by ELISA for anti-TOSV IgG; 47 people (9.5%) were found to be positive. Seroprevalence varied by bioclimatic region and gender. Two hundred and twelve samples, randomly chosen from the same selected population and tested with ELISA, were retested using an IFA for IgG antibodies. An 85% concordance between the IFA and ELISA was obtained (kappa = 0.650). Conclusions: These serological data confirm the circulation of TOSV in different bioclimatic zones in Tunisia where the vector sand flies are found. The detection of IgG against TOSV suggests that the diagnosis of TOSV infection is often neglected, as this virus often causes asymptomatic infections, with only a few patients developing severe illnesses involving neurological manifestations. (C) 2013 International Society for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

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No. of bitstreams: 2 S0103-84782012000800021.pdf: 350265 bytes, checksum: 84259546dc8477eec2b346575b2f6853 (MD5) S0103-84782012000800021.pdf.txt: 32164 bytes, checksum: 68200773f9a28994087ab86cc0016ac4 (MD5) Previous issue date: 2012-08-01 Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) A Erliquiose canina é uma zoonose causada pela Ehrlichia canis, bactéria Gram negativa de distribuição mundial. Alguns cães com erliquiose se tornam portadores assintomáticos enquanto outros desenvolvem uma doença aguda com morte rápida. A apoptose pode ser importante na eliminação de patógenos intracelulares, podendo, nas infecções por Ehrlichia sp. e Anaplasma sp., ocorrer modulação da apoptose celular para prolongar a sobrevivência desses organismos. Para avaliação do papel da apoptose na erliquiose canina, sete cães foram distribuídos em dois grupos. No Grupo inoculado, realizou-se a infecção por via intravenosa com sangue infectado com E. canis (isolado Jaboticabal), sendo realizada a inoculação com PBS estéril nos animais pertencentes ao Grupo Controle. Semanalmente e até 35 dias pós-inoculação, amostras de sangue foram coletadas e submetidas a n-PCR e reação de imunofluorescência (RIFI) para confirmação da infecção. No 36° dia pós-inoculação, os animais foram eutanasiados, sendo as amostras de baço, linfonodo, rim e fígado coletadas e processadas para as técnicas de TUNEL e imunohistoquímica (Bcl-2, Bax). Verificou-se pela n-PCR que os animais inoculados se tornaram positivos para E. canis a partir da segunda semana. Pela RIFI, verificou-se animais com sorologia positiva a partir da terceira semana pós-inoculação. No grupo controle, todos os testes realizados foram negativos para E. canis. Apesar da reação de TUNEL mostrar maior incidência de apoptose no Grupo Inoculado, sendo o baço e os linfonodos os órgãos que apresentaram maior marcação, os resultados da imunohistoquímica para Bcl-2 e Bax indicam que a via intrínseca de apoptose não é importante nos órgãos analisados. Some dogs infected with Ehrlichia canis become asymptomatic while others develop an acute illness followed by quick death. Apoptosis may be an important mechanism for elimination of intracellular pathogens. Also, Ehrlichia sp. and Anaplasma sp. can modulate apoptosis to extend their survival. To evaluate the role of apoptosis in canine ehrlichiosis, 7 dogs were assigned into 2 groups, one with 4 animals inoculated intravenously with blood infected with Ehrlichia canis (Jaboticabal isolate) and a control with 3 dogs, inoculated with sterile PBS. Blood samples were collected weekly and 35 days post-inoculation to confirm the infection by nPCR and immunofluorescence. Thirty-six days after inoculation the animals were euthanized and samples from spleen, lymph nodes, kidney and liver were collected to carry out the TUNEL technique and immunohistochemistry (Bcl-2, Bax). Inoculated animals became positive for E. canis by nPCR already in the second week and by immunofluorescence in the third week after inoculation. The control group showed negative for E. canis in all tests. The TUNEL reaction showed a higher incidence of apoptosis in the inoculated group, with stronger labeling in the spleen and lymph nodes. The results of immunohistochemistry for Bcl-2 and Bax suggest that the intrinsic pathway of apoptosis is not important in the analyzed organs. Universidad del Tolima (UT) Facultad de Medicina Veterinaria y Zootecnia (FMVZ) Universidad de Córdoba (UCO) Facultad de Medicina Veterinaria y Zootecnia Universidade Federal de Minas Gerais (UFMG) Instituto de Ciências Biológicas (ICB) Universidade Estadual Paulista (UNESP) Faculdade de Ciências Agrárias e Veterinárias (FCAV) UNESP FCAV Universidade Estadual Paulista (UNESP) Faculdade de Ciências Agrárias e Veterinárias (FCAV) UNESP FCAV

Cryptosporidium is an intestinal protozoan parasite prevalent in a wide range of mammals. Although it has been recorded in many hosts, its impact on endangered species is poorly understood. Here we present a preliminary study of four populations of the highly threatened spotted souslik (Spermophilus suslicus), living in the westernmost part of the species range. The populations inhabit fragmented habitats and suffer from loss of genetic variation. An IFA test revealed that 35.9% of sampled animals (41/114) was infected with Cryptosporidium and none with Giardia. The prevalence and infection intensity differed among the populations. In areas grazed by cattle it was about 3 folds higher, which suggests a possible transmission route. To the authors best knowledge the present study is the first report of Cryptosporidium infections in S. suslicus.

Background Anaplasma phagocytophilum , the causative agent of granulocytic anaplasmosis, affects several species of wild and domesticated mammals, including horses. We used direct and indirect methods to compare and evaluate exposure to A. phagocytophilum in horses in northern Tunisia. Methods Serum from 60 horses was tested by IFA for antibodies to A. phagocytophilum , and whole blood was tested for A. phagocytophilum 16S rRNA gene using a nested-PCR. To examine the risk of A. phagocytophilum transmission, 154 ticks that had been collected from horses were examined for the presence of A. phagocytophilum by nested-PCR targeting 16S rRNA gene. Results This is the first time that A. phagocytophilum has been detected in horses in Tunisia, with an overall seroprevalence of 40/60 (67%). Six of the seroreactive samples (10%) had an IFA titer of 1:80, 14 (23%) of 1:160, 8 (13%) of 1:320 and 12 (20%) a titer 1 ≥ 640. The seroprevalence revealed no significant regional and sex differences. In contrast, a significant difference was observed between breeds. Eight (13%) of the horses were positive for A. phagocytophilum in the PCR, with no significant breed and age differences. Hyalomma marginatum was a predominant tick species (130/154), and 3 were infected by A. phagocytophilum (a prevalence of 2.3%). The concordance rate of A. phagocytophilum detection between IFA and PCR had a k value of −0.07. Conclusions The results presented in this study suggest that horses infested by ticks in Tunisia are exposed to A. phagocytophilum.

In Brazil, more than 500,000 new cases of malaria were notified in 1992. Plasmodium falciparum and P.vivax are the responsible species for 99.3% of the cases. For adequate treatment, precoce diagnosis is necessary. In this work, we present the results of the traditional Plasmodia detection method, thick blood film (TBF), and the results of alternative methods: Immunofluorescence assay (IFA) with polyclonal antibody and Quantitative Buffy Coat method (QBC)® in a well defined population groups. The analysis were done in relation to the presence or absence of malaria clinical symptoms. Also different classes of immunoglobulins anti-P.falciparum were quantified for the global analysis of the results, mainly in the discrepant results. We concluded that alternative methods are more sensitive than TBF and that the association of epidemiological, clinical and laboratory findings is necessary to define the presence of malaria. Mais de 500.000 casos novos de malária foram notificados no Brasil, em 1992. P.falciparum and P.vivax são as espécies responsáveis por 99,3% dos casos. O diagnóstico precoce é indispensável para início do tratamento adequado. Neste trabalho, apresentamos os resultados do método tradicional para detecção de plasmódios, gota espessa, e os resultados de métodos alternativos estudados: imunofluorescência indireta com anticorpo policlonal anti-P.falciparum e QBC-método, em grupos populacionais bem definidos. A análise dos resultados foi feita em relação à presença ou ausênsia de sintomas clínicos de malária. Também, diferentes classes de imunoglobulinas anti-P.falciparum foram quantificadas para auxiliar na análise global dos resultados, principalmente nos resultados discrepantes. Nós concluímos que os métodos alternativos são mais sensíveis do que a gota espessa e que a associação das informações epidemiológicas, clínicas e laboratoriais é necessária para definir a presença de malária.

To the Editor: Recent outbreaks of Q fever in the Netherlands and the United Kingdom raised public awareness about this ubiquitous bacterial disease known for decades to circulate worldwide (1–4). The disease, which ranges from a self-recovering influenza-like illness to pneumonia and severe meningoencephalitis, myocarditis, or endocarditis, is usually transmitted from animals to humans by airborne particles derived from contaminated feces and birth products. Clinical symptoms develop in only ≈40% of infected humans (1). In ≈1%–2% of these persons, symptoms evolve toward the chronic form of the disease, which can be life-threatening (5). Q fever seroprevalence in the general population in Europe ranges from 2.4% to >30% in some countries in the Mediterranean region (5). Despite improved awareness during the past 3 years and geographic proximity with the Netherlands, few human cases were reported in Belgium (14, 27, and 33 cases in 2007, 2008, and 2009, respectively) (6). A retrospective survey of blood donors in the Netherlands showed a seroprevalence of 2.4% before the start of outbreaks (7). In France, the prevalence of the disease in the Nord-Pas-de-Calais region bordering Belgium is low and accounts for only 0.5% of Q fever cases in France (8). Seroprevalence of the general population in Belgium, although unknown, is thus probably comparable with that in neighboring countries in absence of outbreaks and is not expected to exceed 5%. We report a serologic, epidemiologic, and microbiological Q fever survey conducted in a scouring factory that processed wool and goat hair products in Belgium. No acute Q fever episodes were previously reported by the factory workers. Data on clinical symptoms and risk factors were obtained in face-to-face interviews, and associations with seropositivity were explored by using regression analysis. Airborne dust collected inside the factory during goat hair processing (9) contained 102–103 genome equivalents of Coxiella burnetii, the Q fever agent, per liter of air as estimated by real-time PCR (Laboratoire Service International, Lissieu, France) (Table A1). Sheep wool processing generated less dust and resulted in a C. burnetii air load that never reached 10 genome equivalents/L in our analyses. No information is available about the infectivity or viability of air-suspended C. burnetii in the studied environment. Q fever serologic analysis was conducted by using an in-house ELISA for serum samples from 69 workers obtained annually during 2007–2009. Results of samples from the third year were confirmed in parallel by using an immunofluorescent assay (IFA) (Focus Diagnostics, Cypress, CA, USA) in the reference laboratory in Belgium and with follow-up samples in cases of noninterpretable or suspected serologic profiles. The 3-year cumulative seroprevalence was 50.7% (Table). This high value likely results from occupational exposure inside the factory. However, one cannot exclude that characteristics such as traveling abroad, farming, or living near farm animals might account for part of the seroprevalence. Nevertheless, such characteristics could not be associated with positive serologic results in our epidemiologic analysis. The serologic status of 2 workers (T3 and T42) was compatible with ongoing chronic Q fever as assessed by IFA and ELISA (Table). Another IFA conducted in the reference laboratory in France confirmed a serologic status compatible with chronic Q fever for worker T3 and detected anti–phase I and II IgA in this worker. However, chronic status was not confirmed for worker T42 by this laboratory (Table A1). Profiles of other workers in the cohort were characterized by increased anti–phase II IgG, IgM, or both (9/69, 13%) over a 2-year period. These profiles suggest relapse and may result from continuous exposure to the Q fever agent, which led to reinfection or repeated stimulation of the immune response. Molecular testing did not detect C. burnetii DNA in any blood sample, and clinical examinations did not detect endocarditis in worker T3 as analyzed by positron emission tomography and transthoracic and transesophagian echocardiography. However, infection of tissues other than the heart in this worker cannot be ruled out. Our results indicate high seroprevalence of Q fever among workers at the scouring factory studied. Continuous exposure to the Q fever agent was the likely cause of atypical antibody responses evoking a chronic or relapsing disease in the absence of any clinical symptom. These results indicated the need to analyze paired serum samples and to rely on medical follow-up before establishing a definitive diagnosis. Given the continuous occupational risk to which these workers are exposed, hiring of pregnant women or persons with underlying medical conditions, such as valvulopathy or immunologic depression, should be avoided. Moreover, annual serologic testing should be conducted on all exposed persons to detect any evolution toward the chronic form of the disease, which can be life-threatening. Although less dangerous than anthrax, Q fever is still a highly prevalent occupational disease that affects persons working with animal hairs in industrial environments and commonly referred to as woolsorters (10). Table Serologic results for Q fever in woolsorters, Belgium, 2007–2009*

Aslan, Oznur/0000-0001-5479-3737; Koenhemsi, Lora/0000-0002-4979-170X; Celebi, Bekir/0000-0002-4545-5573 WOS: 000298871500022 PubMed: 21737965 Bartonella hencelae is the causative agent of cat scratch disease (CSD) in humans. Cats are the main reservoir of this bacterium and may infect humans through scratches and bites. The purpose of this study was to determine the B. henselae seroprevalence in cats in Turkey. A total of 298 cats blood samples were collected from six different provinces of Turkey. Sera were tested for the presence of anti-B. henselae IgG antibodies by indirect fluorescent antibody test (IFA). The seroprevalence of B. henselae was 27.9% (83/298) for the cats examined in this study. The seroprevalence of cats by province was significantly higher in Bursa (41.3%), Adana (33.9%), Aydin (27.5%) and Burdur (32.3%) than in Kayseri (17.9%) and Istanbul (12.5%). Statistically significant differences were not observed between cat sexes and living conditions of cats. The results revealed that B. henselae is an important zoonotic pathogen in Turkey.

GAZYAGCI, SERKAL/0000-0002-0043-6942 WOS: 000281923000018 Q fever is a zoonotic disease caused by Coxiella burnetii. This study aimed to detect the relationship of C. burnetii seropositivity between farm animal owners and their animals. Blood serums of 20 farm animal owners, 32 cow and 88 sheep were investigated with indirect Immuno Fluorescent Assay (IFA) using C. burnetii phase I and II antigens. Milk samples of the same animals were tested for C. burnetii by PCR. The serological test results of animals and their owners were compared by statistically methods to reveal the interdependence and correlations. The seropositivities of IgG antibodies against C. burnetii were 90.0% for farm animal owners, 53.1% for cows and 63.6% for sheep. All of the animal owners were consuming dairy products made from their. own animals raw milk. However, as shown by PCR results, none of the tested cows and sheep were responsible for shedding of C. burnetii through their milks. Although, there was no correlation between the shedding pattern and serological results of animals, there was a significant correlation between the serological results of animals and their owners for both phase I and II antigens against C. burnetii. There are statistically important relationships between farm animals and their. owners about phase I and phase II IgG titration levels against C. burnetii. Moreover, there was close dependency between the presence of chronic C. burnetii infections in animals and their owners. On the other hand, serological results of milk samples are not in significant correlation with the serologically dependency of animals and their owners. Kirikkale UniversityKirikkale University [2008/46] This study was supported by the Scientific Research Projects Unit of Kirikkale University (Project no. 2008/46). The researchers would like to express their greatfullness to Dr. Mehmet Tolga Dogru for his guidence and valuable supports in statistical analysis.

This is the first report of serological evidence for bovine immunodeficiency virus (BIV) infection in Argentina. The analysis was performed in 589 dairy bovine sera samples, applying indirect enzyme-linked immunosorbent assay (I-ELISA) using a synthetic antigen (transmembrane peptide, TM) and Immunofluorescent assay (IFA). In this study, 9 dairy herds from 4 Argentinian provinces were evaluated and 12% of the animals tested positive for BIV. Seven of the 9 herds tested were BIV seropositive and the percentage of BIV seropositive animals in the herds ranged from 2% to 42%. Direct detection of BIV provirus applying nested PCR was not conclusive. Antibody detection against bovine leukemia virus (BLV) in all sera was also performed applying immunodiffusion (ID) assay and 59% resulted seropositive. Statistical analysis of the results was carried out and possible evidence of association between BIV and BLV infection was considered. Future studies should be performed including local field isolates strains of BIV., Facultad de Ciencias Veterinarias

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:22:42Z No. of bitstreams: 0 Made available in DSpace on 2014-05-27T11:22:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-12-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) The objective of this study was to develop a direct immunohistochemical assay to detect TCoV antigens in formalin-fixed paraffin-embedded sections prepared from experimentally infected poults. The sections of ileo, ileo-cecal junction and ceca regions from intestine were prepared and submitted to two different primary antibodies, first the non-biotin labeled polyclonal antibody for the indirect method, and second the biotin-labeled polyclonal antibody, both raised against IBV by immunized specific pathogen free chickens. All sections were submitted to immufluorescent assay (IFA), a conventional method, and the results compared. The direct immunohistochemical technique showed a higher frequency of antigen in tissues, especially from the ileo-cecal junction with no difference between results obtained by the conventional method. Finally, the immunofluorescence and all modalities of molecular approaches have been played an important role to the diagnosis and prevention of TCoV infections, although to be precise on infectious disease diagnosis, it is necessary complementary techniques. Here, was standardized the biotin labeled polyclonal antibody as reliable tool to be used as an alternative detection of Turkey Coronavirus. © 2007 Science Publications. Laboratório de Virologia Universidade Estadual Paulista Faculdade de Odontologia, UNESP Campus de Araçatuba, Araçatuba, SP Departamento de Apoio, Produção e Saude Animal, Rua Clóvis Pestana 793, 16.050-680 Araçatuba, SP Fundação de Amparo à Pesquisa do Estado de São Paulo, São Paulo Laboratório de Patologia Animal Universidade Estadual Paulista Faculdade de Odontologia, UNESP Campus de Araçatuba, Araçatuba, SP Laboratório de Virologia Universidade Estadual Paulista Faculdade de Odontologia, UNESP Campus de Araçatuba, Araçatuba, SP Laboratório de Patologia Animal Universidade Estadual Paulista Faculdade de Odontologia, UNESP Campus de Araçatuba, Araçatuba, SP

WOS:000373597200026 The aim of this study was to determine the occurrence of B. burgdorferi sensu lato s.l. antibodies in stray dogs of the urban area of Umuarama Town, localized in the northwest region of Parana state, Brazil. Serum samples from 168 dogs were tested using indirect immunofluorescence assay (IFA) and Western Blot (WB) in order to detect anti-Borrelia burgdorferi s.l. antibodies. The IFI analysis was used as screening test and positive results were confirmed employing the WB technique. Sixty-five (38.69%) of 168 serum samples were positive in the IFI; 54 of them (83.07%) were confirmed by the WB. The overall data analysis confirmed the presence of anti-B. burgdorferi s.l. antibodies in 32.14% (54/168) dogs, what suggests that northwest region of Parana State may constitute a risk area for Lyme disease. Further studies are necessary to determine the epidemiological characteristics of this disease in the region studied. publishersversion published

El Toxoplasma gondii es un parásito intracelular de distribución universal, causante de problemas reproductivos en animales y el hombre. Los animales de sangre caliente constituyen los hospederos intermediarios, presentan quistes tisulares y diseminan la enfermedad al contaminar las pasturas con heces conteniendo ooquistes. Los felinos domésticos y silvestres son los hospederos definitivos. El objetivo del estudio fue determinar la seroprevalencia del T. gondii en borregas en edad reproductiva de una empresa ganadera de la Sierra Central del departamento de Junín, mediante la prueba de Inmunofluorescencia Indirecta (IFI). Se evaluaron 387 sueros de borregas de diferentes edades durante la esquila en marzo del 2002. Se halló una seroprevalencia a T. gondii de 65.9 + 4.7% (255/387). La seroprevalencia se incrementó con la edad de las borregas (p, Toxoplasma gondii is an intracellular parasite of worldwide distribution that causesreproductive disorders in animals and man. Warm-blooded animals constitute theintermediate hosts, get tissue cysts and disseminate the disease when contaminate thepasture with feces containing oocysts. Domestic and wild feline are the definitive hosts.The objective of the study was to determine the seroprevalence of T. gondii in ewesbelonging to a commercial farm located in the Central Highlands of Junín, Peru, using theIndirect Immunofluorescent Antibody Test (IFAT). A total of 387 sera were collected fromewes of various ages at shearing time in March 2002. The detected seroprevalence was65.9 ± 4.7% (255/387). Seroprevalence increased according to age (p

La neosporosis es una enfermedad parasitaria ocasionada por el Neospora caninum, que cursa con problemas reproductivos en el ganado lechero y signos nerviosos en el perro, el cual es el hospedero definitivo. El objetivo del estudio fue determinar la frecuencia de N. caninum en perros de establos lecheros del valle de Lima. Se evaluaron 104 muestras de suero de caninos mayores a 3 meses de edad, provenientes de 23 establos lecheros, mediante la detección de anticuerpos contra N. caninum a través de la prueba de inmunofluorescencia indirecta. El 32.7 ± 9.0% (34/104) de las muestras fueron positivas a N. caninum en una dilución de 1:50; siendo las provincias del Huaura y Huaral las que tuvieron los mayores niveles de seropositividad (58.8%, 10/17). No se hallaron diferencias estadísticas significativas en las variables edad y sexo., Neosporosis is a parasitic disease caused by Neospora caninum that produces nervous signs in dogs, the definite host, and may affect reproduction in dairy cattle. The objective of the study was to determine the frequency of N. caninum in dogs of dairy farms in the Lima valley. Samples from a total of 104 dogs older than 3 months of age from 23 dairy farms, were collected and analyzed by the indirect immunofluorescense test to detect antibodies against N. caninum. The results indicated that 32.7 ± 9.0% (34/104) were positive to N. caninum in a dilution of 1:50. The Huaura and Huaral provinces had the highest proportion of seropositives (58.8%, 10/17). No significant differences were found due to sex and age.

Diagnosis of scrub typhus is difficult because its symptoms are very similar to other acute febrile illnesses, such as leptospirosis, murine typhus, and other viral hemorrhagic fevers. To differentiate scrub typhus from other acute febrile diseases, a rapid and reliable serological diagnosis is important. We have developed a chimeric recombinant antigen cr56 and two other recombinant antigens, r21 and kr56, from various serotypes of Orientia tsutsugamushi. They were tested for the detection of antibodies against O. tsutsugamushi in the patient's serum samples using enzyme-linked immunosorbent assay (ELISA) and dot-blot analyses. As of conventional immunofluorescence assay (IFA), when the mixture of these three recombinant antigens was used, both sensitivity and specificity of the recombinant antigens were increased up to 98% in IgM and IgG at ELISA and dot blotting. Additionally, both sensitivity and specificity by detection of IgM and IgG antibodies at rapid diagnostic test (RDT), using the mixture of three antigens and gold conjugated antibodies, were 99%. Our results suggest the use of mixture of these recombinant antigen proteins in ELISA or RDT is suitable as a diagnostic test for scrub typhus.

Background: The post genomic era of malaria research provided unprecedented insights into the biology of Plasmodium parasites. Due to the large evolutionary distance to model eukaryotes however we lack a profound understanding of many processes in Plasmodium biology. One example is the cell nucleus which controls the parasite genome in a development and cell cycle specific manner through mostly unknown mechanisms. To study this important organelle in detail we conducted an integrative analysis of the P. falciparum nuclear proteome. Results: We combined high accuracy mass spectrometry and bioinformatic approaches to present for the first time an experimentally determined core nuclear proteome for P. falciparum. Besides a large number of factors implicated in known nuclear processes one third of all detected proteins carry no functional annotation including many phylum or genus specific factors. Importantly extensive experimental validation using 30 transgenic cell lines confirmed the high specificity of this inventory and revealed distinct nuclear localization patterns of hitherto uncharacterized proteins. Further our detailed analysis of the core nuclear proteome identified novel protein domains potentially implicated in gene transcription pathways and sheds important new light on nuclear compartments and processes including regulatory complexes the nucleolus nuclear pores and nuclear import pathways. Conclusion: Our study provides comprehensive new insight into the biology of the Plasmodium nucleus and will serve as an important platform for dissecting general and parasite specific nuclear processes in malaria parasites. Moreover as the first nuclear proteome characterized in any protist organism it will provide an important resource for studying evolutionary aspects of nuclear biology.

This study was carried out to evaluate the use of acridine orange fluorescence technique on blood slides as a rapid diagnostic test for tropical theileriosis in comparison with the Giemsa-stained thin blood film technique. Also the indirect fluorescent antibody test has been employed for the serodiagnosis of tropical theileriosis. The study was carried out on 62 young and 48 adult Friesian cattle suffering from clinical tropical theileriosis in Qassim Region, Central Saudi Arabia, during the period from August 2006 to July 2008. For control, blood samples were also obtained from 25 young and 25 adult, clinically healthy, Friesian cattle, selected at random from different dairy farms in Qassim Region. Thin blood films were fixed with methanol and stained with Giemsa and acridine orange and were examined by two independent microbiologists. There was 100% correlation in the interpretation of slides stained with Giemsa and acridine orange both with respect to positivity and negativity, between the two microbiologists. It is concluded that if facilities are available acridine orange is a valuable alternative for screening tropical theileriosis. The method may also have potential value in the diagnosis of Theileria parva, which causes East Coast fever, and also other Theileria species. Results of the present study also showed that IFA test was not found sufficiently sensitive and specific as has been reported earlier. [Vet. World 2011; 4(8.000): 341-344]