Bioactive polysaccharides in different plant parts of Aconitum carmichaelii.

BACKGROUND: Aconitum carmichaelii is an industrially cultivated medicinal plant in China and its lateral and mother roots are used in traditional Chinese medicine due to the presence of alkaloids. However, the rootlets and aerial parts are discarded after collection of the roots, and the non-toxic polysaccharides in this plant have attracted less attention than the alkaloids and poisonous features. In this study, five neutral and 14 acidic polysaccharide fractions were isolated systematically from different plant parts of A. carmichaelii, and their structural features and bioactivity were studied and compared. RESULTS: The neutral fraction isolated from the rootlets differed from those isolated from the lateral and mother roots. It consisted of less starch and more possible mannans, galactans, and/or xyloglucans, being similar to those of the aerial parts. Pectic polysaccharides containing homogalacturonan and branched type-I rhamnogalacturonan (RG-I) were present in all plant parts of A. carmichaelii. However, more arabinogalactan (AG)-II side chains in the RG-I backbone were present in the aerial parts of the plants, while more amounts of arabinans were found in the roots. Various immunomodulatory effects were observed, determined by complement fixation activity and anti-inflammatory effects on the intestinal epithelial cells of all polysaccharide fractions. CONCLUSION: This study highlighted the diversity of polysaccharides present in A. carmichaelii, especially in the unutilized plant parts, and showed their potential medicinal value. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Review of Clinical and Laboratory Diagnostics for Coccidioidomycosis.

Coccidioidomycosis is a fungal disease associated with soil exposure that frequently goes undiagnosed due at least in part to its nonspecific presentation and the lack of clinical suspicion by health care providers. Currently available diagnostics for coccidioidomycosis offer qualitative results that can suffer from low specificity, while semiquantitative assays are labor-intensive and complex and can require multiple days to complete. Furthermore, significant confusion exists regarding the optimal diagnostic algorithms and appropriate usage of available diagnostic tests. This review aims to inform clinical laboratorians and treating clinicians about the current diagnostic landscape, appropriate diagnostic strategies, and future diagnostic directions for coccidioidomycosis, which is expected to become more prevalent due to increased migration into areas of endemicity and climate changes.

Histoplasma capsulatum Complement Fixation and Immunodiffusion Assay Sensitivity in Culture-Confirmed Cases of Histoplasmosis: a 10-Year Retrospective Review (2011 to 2020).

The detection of antibodies against Histoplasma capsulatum remains a frequently relied-on approach to diagnose histoplasmosis. We retrospectively assessed the performances of complement fixation (CF) and immunodiffusion (ID) assays for anti-Histoplasma antibody detection in patients with culture-confirmed histoplasmosis at Mayo Clinic (Rochester, MN) over a 10-year period (2011 to 2020). Among 67 culture-confirmed patients who also had H. capsulatum CF/ID testing ordered, 51 (67.1%) were immunocompromised, 34 (50.7%) had localized disease, and 51 (76.1%) presented with <3 months of symptoms before testing. H. capsulatum CF and/or ID testing was positive in 47 (70.1%) patients, with both assays being positive in 39 cases. CF was positive in 44 (65.7%) patients, with reactivity against both H. capsulatum mycelial and yeast antigens in 30 (68.2%) cases, whereas 11 (25%) and 3 (6.8%) individuals had antibodies to the CF yeast or mycelial antigen only, respectively. H. capsulatum ID was positive in 42 (62.7%) patients, with the presence of the M-band only or the H- and M-bands in 27 (64.3%) and 15 (35.7%) cases, respectively. Among 18 serially tested patients, 12 remained ID and/or CF positive at the final time point (median, 154 days; range, 20 to 480 days). Serial CF testing showed that antibodies to the mycelial antigen serorevert to negative more frequently (6/11) than antibodies to the yeast antigen (2/13). There was no statistically significant difference in antibody positivity relative to patient immune status, degree of disease dissemination, or symptom duration. Serologic testing remains a valuable asset to support the diagnosis of histoplasmosis, particularly when direct detection methods fail to identify an infection.

Coccidioidomycosis Complement Fixation Titer Trends in the Age of Antifungals.

Coccidioidomycosis is associated with a broad spectrum of illness severity, ranging from asymptomatic or self-limited pulmonary infection to life-threatening manifestations of disseminated disease. Serologic studies before the widespread availability of antifungals established current understanding of serologic kinetics and dynamics. Chart histories and complement fixation (CF) titer trends were analyzed for 434 antifungal-treated coccidioidomycosis patients, who were classified by three infectious disease physicians as having either pulmonary uncomplicated coccidioidomycosis (PUC) (n = 248), pulmonary chronic coccidioidomycosis (PCC) (n = 64), disseminated coccidioidomycosis (DC) not including meningitis (n = 86), or coccidioidal meningitis (CM) (n = 36). The median maximal CF titers were 1:4 for PUC patients, 1:24 for PCC patients, 1:128 for DC patients, and 1:32 for CM patients. Approximately 25.4% of PUC patients, 6.2% of PCC patients, 2.3% of DC patients, and 8.3% of CM patients did not develop detectable titers during the study period. Maximal titers developed a mean of 31 days (95% confidence interval [CI], 13 to 50 days) after initial serologic positivity, with no significant differences between groups. Serologic recurrence occurred in 9% of PUC patients, 36% of PCC patients, 50% of DC patients, and 52% of CM patients. Median titer improvement rates were 91 days/dilution for PUC patients, 112 days/dilution for PCC patients, 136 days/dilution for DC patients, and 146 days/dilution for CM patients. Receiver operating characteristic (ROC) analysis revealed that CF testing retains moderate classification value for disseminated infections (area under the curve [AUC], 0.82 [95% CI, 0.78 to 0.87]) and complicated infections (AUC, 0.82 [95% CI, 0.77 to 0.86]). A suitable cutoff value for complicated infections is ≥1:32. Findings update serologic parameters that are relevant for clinical assessment of coccidioidomycosis patients in the triazole era.

Clinical risk stratification of paediatric renal transplant recipients using C1q and C3d fixing of de novo donor-specific antibodies.

INTRODUCTION: We have previously shown that children who developed de novo donor-specific human leukocyte antigen (HLA) antibodies (DSA) had greater decline in allograft function. We hypothesised that patients with complement-activating DSA would have poorer renal allograft outcomes. METHODS: A total of 75 children developed DSA in the original study. The first positive DSA sample was subsequently tested for C1q and C3d fixing. The primary event was defined as 50% reduction from baseline estimated glomerular filtration rate and was analysed using the Kaplan-Meier estimator. RESULTS: Of 65 patients tested, 32 (49%) and 23 (35%) tested positive for C1q and C3d fixing, respectively. Of the 32 C1q-positive (c1q+) patients, 13 (41%) did not show concomitant C3d fixing. The mean fluorescence intensity values of the original immunoglobulin G DSA correlated poorly with complement-fixing positivity (C1q: adjusted R 2 0.072; C3d: adjusted R 2 0.11; p < 0.05). C1q+ antibodies were associated with acute tubulitis [0.75 ± 0.18 (C1q+) vs. 0.25 ± 0.08 (C1q-) episodes per patient (mean ± standard error of the mean; p < 0.05] but not with worse long-term renal allograft dysfunction (median time to primary event 5.9 (C1q+) vs. 6.4 (C1q-) years; hazard ratio (HR) 0.74; 95% confidence ratio (CI) 0.30-1.81; p = 0.58]. C3d-positive (C3d+) antibodies were associated with positive C4d histological staining [47% (C3d+) vs. 20% (C3d-); p = 0.04] and with significantly worse long-term allograft dysfunction [median time to primary event: 5.6 (C3d+) vs. 6.5 (C3d-) years; HR 0.38; 95% CI 0.15-0.97; p = 0.04]. CONCLUSION: Assessment of C3d fixing as part of prospective HLA monitoring can potentially aid stratification of patients at the highest risk of long-term renal allograft dysfunction.

The experience of contagious bovine pleuropneumonia ring trials in Botswana.

The Botswana National Veterinary Laboratory (BNVL) has conducted ring trials (proficiency testing) on an annual basis since 2010. Proficiency testing is carried out to evaluate the ability of veterinary laboratories to perform serological complement fixation tests (CFTs) and molecular polymerase chain reaction (PCR) tests for the diagnosis of contagious bovine pleuropneumonia (CBPP). In this paper, the authors discuss the experience gained and the lessons learned in coordinating these ring trials over a period of six years, from 2010 to 2015.The number of participating laboratories increased from five in 2010 to 11 in2015. Their performance also improved over this period. The proportion of unsatisfactory results decreased from 40% to 10% for serological testing, while questionable results decreased from 60% to 10%. The proportion of unsatisfactory results for the molecular test decreased from 33% to 0%. Systematic errors (i.e. technical errors or imperfect experimental design) were the principal causes of questionable and unsatisfactory results. An analysis of responses from customer satisfaction surveys conducted annually since 2013 provided valuable information that enabled BNVL to redesign the programme in 2014 and 2015 to improve the overall quality of the proficiency testing programme. Among the changes made were sending freeze-dried sera for CFTs and DNA for PCR instead of sera and liquid cultures.

Le Laboratoire national vétérinaire du Botswana (BNVL) effectue depuis 2010 des essais comparatifs inter-laboratoires à intervalles d'un an. Ces essais d'aptitude inter-laboratoires visent à évaluer la capacité des laboratoires vétérinaires du pays à réaliser efficacement le test de fixation du complément (FC) ainsi qu'un test moléculaire (l'amplification en chaîne par polymérase ou PCR) en vue du diagnostic de la péripneumonie contagieuse bovine (PPCB). Les auteurs décrivent l'expérience acquise et les enseignements tirés lors de la coordination d'essais comparatifs inter-laboratoires sur une période de six ans (de 2010 à2015). Le nombre de laboratoires participants est passé de cinq en 2010 à onze en 2015. Les performances des laboratoires se sont également améliorées au cours de cette période. Le pourcentage de résultats sérologiques non concluants est passé de 40 % à 10 % tandis que la part des résultats douteux est passée de60 % à 10 %. Pour ce qui concerne la technique moléculaire, la proportion de résultats non concluants est passée de 33 % à 0 %. Les résultats douteux et non concluants étaient essentiellement dus à des erreurs systématiques (c'est-à-dire à des erreurs techniques ou imputables à une conception imparfaite du protocole de test). Une analyse des réponses fournies aux enquêtes de satisfaction réalisées chaque année depuis 2013 a fourni des informations précieuses qui ont permis au BNVL de corriger le protocole du programme en 2014 et en 2015 afin d'améliorer la qualité globale du programme d'essais d'aptitude. Les principales modifications introduites ont été le recours à des sérums lyophilisés pour le test de FC et l'utilisation d'ADN lyophilisé pour les PCR au lieu des sérums et des cultures en milieu liquide.

El Laboratorio Veterinario Nacional de Botsuana (BNVL) lleva realizando pruebas interlaboratorios anuales (pruebas de competencia) desde 2010. Las pruebas de competencia sirven para evaluar la aptitud de un laboratorio veterinario para efectuar pruebas serológicas (la de fijación del complemento, FdC) y moleculares (la de reacción en cadena de la polimerasa, PCR) de diagnóstico de la perineumonía contagiosa bovina. Los autores examinan la experiencia adquirida y las enseñanzas extraídas con la coordinación de estas pruebas interlaboratorios durante el sexenio que va de 2010 a 2015. El número de laboratorios participantes pasó de 5 en 2010 a 11 en 2015. Su eficacia también mejoró durante esos años: en el caso de la técnica serológica, el porcentaje de resultados insatisfactorios pasó de un 40% a un 10 y el de resultados cuestionables de un 60% a un 10%. En el caso de la técnica molecular, el porcentaje de resultados insatisfactorios pasó de un 33% a un 0%. Los resultados insatisfactorios o cuestionables se deben principalmente a errores sistemáticos (esto es, errores técnicos o concepción imperfecta de la prueba). Al analizar las respuestas a los cuestionarios de satisfacción del cliente realizados anualmente desde 2013 se obtuvo información de gran interés, que sirvió al BNVL para remodelar el programa de pruebas de competencia en 2014 y 2015 a fin de mejorar su calidad general. Entre los cambios introducidos está el envío de sueros liofilizados para la prueba de FdC y de ADN liofilizado para la PCR, en lugar de sueros y cultivos líquidos.

[Diagnostic procedure after abortions in sows after simultaneous infection with leptospira and chlamydia]. / Diagnostische Abklärung von Aborten bei Zuchtsauen nach Leptospiren- und Chlamydieninfektion.

INTRODUCTION: In a farrowing farm 2 first parity sows aborted on day 95 and day 110 of gestation due to an infection with leptospira and chlamydia. The double infection was diagnosed by PCR examination of abortion material. Serum samples of both sows and additional 8 sows taken three weeks after abortions were sent to two different labs for serological examination for antibodies against leptospira and chlamydia using a microagglutination test and a complement fixation test, respectively. In both labs the tests for antibodies against chlamydia were negative. Titers against diverse leptospira serovars varied between both labs and were low, so that they were not indicative for the involvement of the two pathogens regarding abortion. This case report indicates the diagnostic difficulties of direct and indirect detection methods for leptospira and chlamydia to assess the impact of these pathogens on observed reproductive failure.

Immunological and biochemical characterisation of 7ap, a short protein translated from an alternative frame of ORF7 of PRRSV.

Sequence analysis revealed a short alternative open reading frame (ORF) named ORF7a within the nucleocapsid gene of genetically divergent porcine reproductive and respiratory syndrome virus (PRRSV) genomes. Alignment of the corresponding protein sequences (named 7ap) revealed substantial heterogeneity among 7aps of different genotypes, though all of them are predicted to be positively charged. Green fluorescent protein and FLAG fusion constructs of ORF7a of the HU-14432/2011 PRRSV demonstrated that 7ap is expressed. 7ap of HU- 14432/2011 (Hu7ap) was synthesised chemically, and ELISA experiments revealed that Hu7ap binds strongly to mammalian IgGs. Protein-protein gel retardation assays and complement fixation inhibition suggest that 7aps bind to the CH2 domain of the IgG(Fc) fragment. Cellular localisation and immunological characteristics of PRRSV 7ap may indicate multiple functions including nuclear and cytoplasmic over-tuning of normal cellular processes and immunosuppression.

Isolation, serological and molecular methods in screening of Burkholderia mallei in East Azerbaijan province, Iran.

Glanders caused by Burkholderia mallei is one of the most dangerous zoonotic diseases in solipeds. Clinical diagnosis of this disease in its early stages in horses, is difficult. This study investigated serological and molecular identification of B. mallei in East Azerbaijan province. In the third and fourth quarters of 2020, throughout 2021, and in the first and second quarters of 2022, the complement fixation test (CFT) was performed on 350 horses. The malleination was used to confirm the positive CFT cases. Blood samples were taken for culture and for preparing serums to perform the enzyme-linked immunosorbent assay (ELISA). Deep eye discharge, nostril, cutaneous ulcers and lymph fluid swabs were cultured, and polymerase chain reaction (PCR) was carried out. Eleven horses were CFT-positive. Based on the malleination on the 11 horses, six were affected by glanders, five were not affected (false positive), and one horse was CFT-negative despite exhibiting clinical signs. It was positive by malleination, ELISA and PCR. A total number of seven positive cases of glanders were diagnosed. The B. mallei could not be isolated, but the Burkholderia cepacia complex was isolated in one case. Except for three cases, the results of the CFT, mallein and ELISA tests were consistent. The amount of confidence interval was 95.00%. It is suggested that ELISA could be used as a complement to CFT in screening and, if positive results are observed in one of the tests, the entire herd must be examined more accurately using the mallein and western blot confirmatory tests.

Seasonal Testing, Results, and Effect of the Pandemic on Coxsackievirus Serum Studies.

Coxsackieviruses (CVs) are common causes of infections and can be life-threatening. Unfortunately, rigorous studies guiding the clinician in interpreting CV serum antibody titer testing is lacking. To explore the epidemiology of circulating CVs and the serological test utility in aiding diagnosis of CV infections in our community, we obtained results of CV immunologic diagnostic tests between 2018 and 2022 from a regional healthcare database. For CV type A, rare individuals had positive CF (complement fixation) tests whereas all 16 individuals with IFA testing showed at least one positive serotype. For CV type B CF testing, 52.2% of 222 patients had at least one serotype positive, with B5 being most common and also the most common with higher titers (14.8% with ≥1:32). We found a significant reduction in seropositivity rate during the pandemic in 2020 compared to 2018, which continued through 2022 (OR: 0.2, 95% CI: 0.08-0.49, p-value < 0.001). During the pandemic, the seasonal pattern of positive tests varied from the pre-pandemic pattern. Testing for CVs was increased after the first year of the pandemic. Overall, the variability by month and seasonal change in our data support that CF testing can be used to identify recent CVB infection.

Evaluation of three serological tests for the diagnosis of Brucella suis in dogs using Bayesian latent class analysis.

Brucella suis infection of dogs is an emerging issue worldwide requiring specific management to address zoonotic risks and animal welfare concerns. Diagnosis in dogs is routinely based on serological testing, but to date these tests have only been validated for use in production animal species and humans. This study aimed to assess the diagnostic performance of three commonly used serological tests in dogs. Canine sera (n = 989) were tested with the Rose Bengal rapid plate agglutination test (RBRPT), the complement fixation test (CFT) and a competitive ELISA (C-ELISA). Diagnostic test performance was evaluated using a three test, two population Bayesian latent class analysis accounting for conditional dependence between the three tests. Positive and negative predictive values (PPV, NPV) were calculated for a range of expected prevalence estimates for the individual tests and test combinations interpreted in series and parallel. The RBRPT showed the highest individual Se of 0.902 (95 % posterior credible interval [PCI] 0.759-0.978) and the CFT the highest individual diagnostic specificity (Sp) of 0.914 (95 % PCI 0.886-0.946). The C-ELISA had marginally the best overall diagnostic performance (Youden's index = 0.807). The CFT and the C-ELISA interpreted in parallel returned the highest combined Se and Sp (0.988 and 0.885, respectively). All tests returned NPVs of > 0.982 in 2-8 % prevalence settings. Series interpretation of the three-test combination as well as the two-test combinations of the RBRPT and the C-ELISA and the CFT and the C-ELISA produced a PPV of 0.502 when the estimated prevalence was 8 %. While all tests are suitable for the detection of B. suis antibodies in dogs, they should not be interpreted in isolation as their diagnostic value is dependent on the pre-test probability of the disease. As such they are useful tools for the diagnosis of B. suis in dogs when exposure, history and clinical presentation indicate a risk of brucellosis.

Pediatric coccidioidomycosis in central California: a retrospective case series.

BACKGROUND: Coccidioidomycosis, an endemic fungal infection seen throughout the southwestern United States, is not well described in children. METHODS: We performed a retrospective observational study of all children admitted to Children's Hospital Central California with coccidioidomycosis from 1 January 2010 to 1 September 2011. RESULTS: Thirty-three children, aged 6 months to 17 years, were hospitalized during the study period. These included patients with pneumonia (n = 28), pleural effusion (n = 13), pleural empyema (n = 4), lung abscess (n = 7), pericarditis (n = 2), osteomyelitis (n = 5), meningitis/cerebritis (n = 2), and vocal cord infection (n = 1). Mediastinitis, with radiographic evidence of purulence and necrotic/abscessed lymph nodes in the mediastinum, was present in 7 patients (21%) and tended to occur more often in younger children (median age, 3 years [range, 0.5-11 years] vs 7 years [range, 0.6-17 years] for non-mediastinitis patients; P = .10). Seven patients were admitted to the intensive care unit and 10 required surgical intervention. One patient died of meningitis. Hospitalizations were longer for patients with mediastinitis (median, 130 days [range, 58-200 days] vs 43 days [range, 3-273 days for non-mediastinitis patients]; P < .01) and those with maximum coccidioidal complement fixing antibody titers ≥1:128 (median, 174 days [range, 53-273 days] vs 33 days [range, 3-200 days] for those with maximum titers <1:128; P < .01). CONCLUSIONS: Coccidioidomycosis causes a substantial disease burden in the children of central California. Mediastinitis is common and tends to occur in younger children. Patients with mediastinitis or elevated coccidioidal complement fixation titers require longer hospitalizations. Further research is needed on the prevention and treatment of this disease.

Improving a Complement-fixation Test for Equine Herpesvirus Type-1 by Pretreating Sera with Potassium Periodate to Reduce Non-specific Hemolysis.

Non-specific hemolysis has often been observed during complement-fixation (CF) tests for equine herpesvirus type-1 (EHV-1), even when the sera have virus-specific CF antibodies. This phenomenon has also been reported in CF tests for various infectious diseases of swine. We found that the sera from 22 of 85 field horses (25.9%) showed non-specific hemolysis during conventional CF testing for EHV-1. Because pretreatment of swine sera with potassium periodate (KIO4) improves the CF test for swine influenza, we applied this method to horse sera. As we expected, horse sera treated with KIO4 did not show non-specific hemolysis in the EHV-1 CF test, and precise determination of titers was achieved.

Serological and Bacteriological Surveillance of Glanders Among Horses in Central Region of Iran.

Glanders is the oldest and very contagious disease among horses caused by Burkholderia mallei. The disease occurs as a chronic form in horses. Hence, because of the prolonged shedding, numerous horses can potentially get infected by one horse with glanders. Glanders is endemic in Iran and this causes occasional occurrence in horse population of the country. The aim of this study was to determine the incidence of B.mallei infection in horses in two central provinces of Iran. A total of 517 serum samples were collected from stable horses in Tehran and Alborz provinces. Among the studied horses, seven presented fever, anorexia, dyspnea, subcutaneous abscesses, nasal and cutaneous discharges, emaciation, and lymphadenopathy. Nasal and ocular discharges and subcutaneous abscesses were sampled for bacterial culture and PCR. The sera were examined by means of complement fixation test (CFT) and indirect enzyme-linked immunosorbent assay (iELISA). Seropositive cases were further examined by Mallein test. The results derived from the present study indicated that only 1.35% of the studied horses were positive in CFT, iELISA and Mallein test, of which only in 42.85% B.mallei was successfully cultured on blood agar and glycerinated nutrient media and confirmed by PCR. Periodic serological tests along with quarantine can benefit reduction of the occurrence of the disease in horses in Iran.

European Inter-Laboratory Proficiency Test for Dourine Antibody Detection Using the Complement Fixation Test.

Dourine is a sexually transmitted parasitic disease affecting equids. Its causative agent is referred to as Trypanosoma equiperdum and the prescribed serodiagnosis method is the complement fixation test (CFT). In the context of our European Reference Laboratory mandate for equine diseases (excluding African horse sickness), we organised dourine CFT inter-laboratory proficiency tests (ILPTs) in 2015, 2018 and 2022 to evaluate the performance of the European Union network of National Reference Laboratories (NRLs) for dourine. ILPT panels were composed of horse sera with or without antibodies against Trypanosoma spp. originating from non-infected, immunised or experimentally infected horses. Twenty-two NRLs participated in at least one of the three sessions. In 2015, 2018 and 2022, the percentage of laboratories obtaining 100% of the expected results was 57, 90 and 80, respectively. These dourine CFT ILPTs showed the benefits of standardising the method's detection limit and underlined the constant need to evaluate NRLs to improve the network's performance. These results also argue in favour of the need for a representative bio-bank to improve the representativeness of ILPT samples and to allow the adoption of alternative serological methods for international surveillance of dourine.

Study on small ruminant brucellosis and owners awareness in two selected districts of southern region, Ethiopia.

INTRODUCTION: Brucellosis is one of the infectious diseases that has the greatest impact on the productivity of sheep and goats. A cross-sectional study followed by a simple random sampling technique was used to investigate the seroprevalence of brucellosis (Rose Bengal plate test; RBPT and complement fixation test; CFT) in small ruminants and its related risk variables from November 2019 to June 2020 in Kolme and Abala Abaya districts. A questionnaire was also given to owners to assess their existing knowledge of the disease. RESULT: Using the RBPT and CFT, 28 (4.1%) and 23 (3.33%) of the 690 animals were found to be seropositive for brucellosis, respectively. In this study, the seroprevalence of brucellosis detected in the Kolme district (5.3%) was greater when compared to Abala Abaya (1.0%). The odds of Brucella infection were greater for goats (odds ratio [OR] 6.0, 95% confidence interval [CI] 16 0.8-44.9) than for sheep. The odds of adult animals (OR 0.05, 95% CI 0.03-0.07) being positive for brucellosis was higher than young animals. A statistically significant difference in the seropositivity of brucellosis was detected in univariate logistic regression among districts, different age groups, herd size, parity number, and reproductive health problems except for species and sex, but in multivariate logistic regression, only reproductive health problems were revealed a statistically significant difference. Out of 138 families, 100% of respondents were unaware of brucellosis, 94.5% drink raw milk, and 74% handle animals with retained fetal membranes with their bare hands. CONCLUSION: This study showed that brucellosis was a widely spread disease in the study areas and poses a substantial public health danger. To reduce the spread of the disease in small ruminants, public health risks, and economic losses, stringent vaccination application and awareness of personal hygiene are critical.

Brucella abortus Strain RB51 Administered to Prepubescent Water Buffaloes, from Vaccination to Lactation: Kinetics of Antibody Response and Vaccine Safety.

Brucella RB51 is a live modified vaccine. Its use in water buffalo has been proposed using a vaccination protocol different to that used for cattle, but knowledge of the long-term effects of RB51 vaccination in this species remains incomplete. The aim of the study was to evaluate the safety and kinetics of antibody responses in water buffaloes vaccinated according to the protocol described for the bovine species in the WOAH Manual, modified with the use of a triple dose. Water buffaloes were vaccinated with the vaccine RB51. A booster vaccination was administered at 12 months of age. When turning 23-25 months old, female animals were induced to pregnancy. RB51-specific antibodies were detected and quantified using a CFT based on the RB51 antigen. Vaccinated animals showed a positive serological reaction following each vaccine injection, but titers and the duration of the antibody differed among animals. For 36 weeks after booster vaccination, the comparison of CFT values between vaccinated and control groups remained constantly significant. Afterwards, antibody titers decreased. No relevant changes in antibody response were recorded during pregnancy or lactation. In conclusion, results indicated that the vaccination schedule applied is safe and allows for vaccinated and unvaccinated controls to be discriminated between for up to 8 months after booster vaccination.

Serologic Rebound after Stopping Azoles for Primary Pulmonary Coccidioidomycosis: A Case-Controlled Observational Study.

BACKGROUND: We sought to characterize the outcomes of patients with primary pulmonary coccidioidomycosis whose post-treatment complement fixation (CF) titer increased by more than 2 dilutions (serologic rebound) after discontinuation of antifungal treatment. METHODS: We conducted a retrospective chart review of patients with primary pulmonary coccidioidomycosis and identified immunocompetent, non-pregnant adults who received antifungal treatment and then experienced a serologic rebound after treatment discontinuation. We compared these to matched controls similarly treated who did not have serologic rebound. RESULTS: Fifty-eight patients experienced serologic rebound. Thirty (52%) of these were associated with symptoms. Nine were associated with radiographic progression. The median time to serologic rebound was 3.5 months. Antifungal treatment was reinitiated in 37 (63.7%) patients. Four of the 58 (6.9%) with rebounded serology subsequently developed extra-thoracic dissemination. Compared with matched controls, patients with rebounded serology were more likely to have post-treatment symptoms, reinitiation of antifungal therapy, and a longer duration of clinical follow-up. However, they were not more likely to experience extra-thoracic dissemination. CONCLUSION: Serological rebound, manifested in at least 2-dilution rise of CF titer following antifungal treatment of primary pulmonary coccidioidomycosis, was uncommon, but resulted in longer clinical follow-up. Continued monitoring of such patients is important to identify the patients who develop subsequent symptoms, as well as extra-thoracic dissemination.

Clinical investigation and management of Brucella suis seropositive dogs: A longitudinal case series.

BACKGROUND: Brucellosis in dogs caused by Brucella suis is an emerging zoonotic disease. OBJECTIVES: To document clinical characteristics, serology, microbiology, and clinical response to treatment in B. suis-seropositive dogs. ANIMALS: Longitudinal study of 27 privately-owned dogs. Dogs that tested positive by serology, culture, or real-time polymerase chain reaction (qPCR) were included in the study. METHODS: Clinical (physical examination and imaging) and laboratory (serology, hematology, serum biochemistry, and qPCR or culture) assessments were made at baseline and after approximately 3, 6, 12, and 18 months. RESULTS: Dogs were followed for 10 895 dog days, with 17/27 dogs completing the 18-month follow-up. Ten dogs had signs consistent with brucellosis before enrollment (n = 4), at baseline (n = 2) or during follow-up (n = 6), with 2 dogs experiencing relapse of historical signs. Antibody titers persisted for the duration of follow-up in 15/17 dogs (88%). Radiographic (n = 5) and ultrasound (n = 11) findings, of variable clinical relevance, were observed. Brucella DNA and organisms were detected in 3 dogs, all of which had clinical signs, including in the milk of a bitch around the time of whelping. Brucella DNA was not detected in blood (n = 92 samples), urine (n = 80), saliva (n = 95) or preputial swabs (n = 78) at any time during follow-up. Six dogs underwent treatment, all of which achieved clinical remission although remission was not reflected by decreasing antibody titers. CONCLUSIONS AND CLINICAL IMPORTANCE: Most dogs with B. suis infections have subclinical infections. Serology is poorly associated with clinical disease. Excretion of organisms appears rare except in whelping bitches. Clinical management using antibiotics with or without surgery is recommended.

Seroprevalence of brucellosis in communal and smallholder cattle farming in North West Province, South Africa.

Brucellosis is an important bacterial zoonosis responsible for considerable economic losses in livestock and health-related burden worldwide. The objective of this study was to determine the seroprevalence of brucellosis in communal and smallholder cattle farming in four districts of the North West province of South Africa (Dr Ruth Segomotsi Mompati, Ngaka Modiri Molema, Bojanala platinum and Dr Kenneth Kaunda districts). Seven hundred and seventy blood samples from farmed animals (n = 378) and abattoir-slaughtered animals (n = 392) were collected. In addition, milk samples (n = 22) were collected from lactating farmed cows. Rose Bengal test (RBT), complement fixation test (CFT) and milk ring test (MRT) were used to detect antibodies against Brucella species. The RBT showed a seroprevalence of 2% at 95% confidence interval (CI: 1.35-3.35), CFT confirmed an overall seroprevalence of 1.95% (95% CI: 1.14-3.12) for all four districts sampled. Although the seroprevalence of brucellosis was found to be low, the possibility of undetected cases of the disease could not be ruled out. Overall, the findings of this study confirmed that brucellosis is endemic in the surveyed areas of the North West province of South Africa.Contribution: The outcome of this study will contribute to the National Brucellosis Project organised by the Department of Agriculture, Land Reform and Rural Development (2016-2026) to assist in the effective implementation of the disease control measures with a view to prevent further outbreaks in the country's cattle population.