scholarly journals Seroprevalence and Molecular Detection of Bovine Brucellosis

Author(s):  
Raghavendra Prasad Mishra ◽  
Udit Jain ◽  
Barkha Sharma ◽  
Kaushal Kusum ◽  
Neha Singh

Background: Brucellosis is one of the major zoonotic problems that exist worldwide. Brucellosis is clinically characterized by metritis, mastitis, repeat breeding, abortion in the last trimester of pregnancy, retention of placenta and reduced milk production in the female whereas epididymitis, orchitis and sterility in male. In humans can be highly variable, ranging from nonspecific, flu-like symptoms to undulant fever, arthritis, orchitis and epididymitis. Methods: A total of 567 bovine serum samples was taken from four districts of Brij region of UP. All the samples were processed to detection of prevalence of brucellosis by RBPT, STAT ELISA and confirmation of genes bcsp31, 16SrRNA, omp2 and IS711 by PCR. Result: The prevalence of brucellosis was found to be 07.93% (31/391), 08.69% (34/391) and 10.74% (42/391) shows positive by RBPT, STAT and I- ELISA respectively. In buffalo Out of 176 tested serum sample the seroprevalence was found to be 09.66% (17/176), 10.79% (19/176) and 12.5% (22/176) positive by RBPT, STAT and I- ELISA respectively. Out of 567 samples 18 were positive for Brucella genus specific gene. The higher prevalence of the disease in this region increases the risk of zoonotic transmission and it implies a serious threat to the human population as well as the huge impact on economy due to loss of productivity as well as loss of livestock population.

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Fevziye Burcu Sirin ◽  
Hakan Korkmaz

AbstractIn the present study we report a case of thyroglobulin (TGB) measurement interference in a total thyroidectomized and radio-ablated 61-year old woman with papillary thyroid cancer. We investigated possible interference in the measurement of TGB due to discordant TGB in relation to clinical condition during the follow-up period. Serum TGB was measured with the chemiluminescence method using Beckman Coulter Unicel DxI 800 instrument. To investigate possible interference in TGB measurement serial dilutions, polyethylene glycol precipitation (PEG), treatment with heterophile blocking tube (HBT), rheumatoid factor level determination and retesting of TGB with an alternative method were performed. Serial dilutions of the serum sample revealed linearity but a remarkable decrease in TGB in the patient’s serum samples post PEG and post HBT treatments. Also, TGB results under functional sensitivity level obtained with a different method suggested that TGB interference developed due to heterophile antibody presence in the serum sample. The patient had unnecessarily undergone expensive imaging techniques, and invasive procedures such as lymph node fine needle aspiration biopsy, before the analytical interference was suspected by the clinician. This report illustrates the importance of early communication and close collaboration between clinicians and laboratorians in order to avoid unnecessary clinical intervention.


2016 ◽  
Vol 37 (5Supl2) ◽  
pp. 3413
Author(s):  
Erivânia Camelo de Almeida ◽  
Aderaldo Alexandrino Freitas ◽  
Késia Alcântara Queiroz Pontual ◽  
Marcília Maria Alves Souza ◽  
Marcos Amaku ◽  
...  

This study was conducted to characterize the epidemiology of bovine brucellosis in the state of Pernambuco, Brazil. The state was divided into three regions, and in each region, approximately 300 properties were randomly sampled. From these selected properties, a pre-established number of animals were randomly selected and blood serum samples were obtained. A total of 3,901 animals were selected from 900 properties. For each selected property, an epidemiological questionnaire was administered to assess the type of farming, the animal husbandry practices and the sanitary practices that could be associated with the presence of brucellosis infection. The testing protocol consisted of screening the samples with a buffered acidified plate antigen test and retesting the positive samples with a complement fixation test (CF). One positive animal was enough to define an infected herd. The prevalence rates of infected herds and animals in the state were 4.5% [3.2; 6.4%] and 1.4% [0.7; 2.7%], respectively. By region, the prevalence rates of infected herds and animals, respectively, were as follows: Zona da Mata, 3.3% [1.8; 6.1%] and 1.7% [0.5; 3.0%]; Agreste, 7.4% [4.9; 10.9%] and 1.9% [0.8; 3.0%]; and Sertão, 1.3% [0.5; 3.5%] and 0.7% [0.0; 1.6%]. Flooded pastures (OR = 2.86 [1.37; 6.42]) and the presence of 13 or more females in the herd (3rd quartile) (OR = 2.65 [1.19; 5.89]) were identified as risk factors. The existence of veterinary care emerged as a protective factor against bovine brucellosis in the state of Pernambuco (OR = 0.24 [0.10; 0.58]).


2016 ◽  
Vol 37 (5Supl2) ◽  
pp. 3403 ◽  
Author(s):  
Inácio José Clementino ◽  
Ricardo Augusto Dias ◽  
Marcos Amaku ◽  
Fernando Ferreira ◽  
Evelise Oliveira Telles ◽  
...  

This study was performed to characterize the epidemiological status of brucellosis in the State of Paraíba, Brazil. The State was divided into three regions. Herds were randomly sampled in each region and a pre-established number of animals were sampled in each of these herds. A total of 3,489 serum samples from 674 herds were collected. In each herd, an epidemiological questionnaire was conducted. This questionnaire focused on herd traits, as well as husbandry and sanitary practices that could be associated with the risk of infection. The serum samples were screened for antibodies against Brucella spp. by the Rose-Bengal Test (RBT), and all positive sera were confirmed by the 2-mercaptoethanol test (2-ME). The herd was considered positive if at least one animal had positive results for both the RBT and the 2-ME test. The prevalence rates of infected herds and animals in the State were 4.6% [3.2-6.5%] and 2.5% [1.1-3.9%], respectively. The prevalence rates of infected herds and animals in the regions were, respectively: region 1, 3.2% [1.5-6.6%] and 1.7% [0.5-5.7%]; region 2, 2.2% [0.9-5.2%] and 0.7% [0.3-1.7%]; and region 3, 7.9% [5.0-12.2%] and 3.2% [1.6-6.3%]. The risk factor (odds ratio, OR) associated with the presence of the infection was Zebuine as the predominant breed (OR=12.30 [1.32-114.64]).


2005 ◽  
Vol 128 (2) ◽  
pp. 436-441 ◽  
Author(s):  
Donna M. Meyer ◽  
Adam Tillinghast ◽  
Nevan C. Hanumara ◽  
Ana Franco

This paper describes an experimental method, bio-ferrography, to separate ultrahigh molecular weight polyethylene (UHMWPE) wear debris, generated in hip simulators, from bovine serum lubricating fluid. A total of 54 experiments were performed in which an enzyme digestion “cocktail” was developed and used to clean the bovine serum samples of extraneous sugars, proteins, and lipids that interfere with the UHMWPE particle separation. Erbium chloride was used to marginally magnetize particles in the fluid prior to passing through the ferrographic device. The particles were captured and separated from the fluid by traversing the treated serum across a magnetic gap of a bio-ferrograph. Morphology of the captured and separated wear debris was compared with particles from samples of fluid filtered through a paper sieve arrangement with pores of 0.05micrometers in diameter. The UHMWPE wear debris collected using the described experimental method, were found to be between 0.1 and 20micrometers in diameter with spherical and pill-shaped particles. The filtered UHMWPE particles were in the same size range as the debris separated using bio-ferrography. To show that the experimental method captured UHMWPE particles, the spectra of the chemical composition of UHMWPE from an acetabular cup insert of a hip implant and of UHMWPE particles separated using bio-ferrography were compared and found to be the same. To further demonstrate that polyethylene could be captured and separated through the experimental method, manufactured polyethylene microspheres in the diameter range of 3-45micrometers, were captured and separated using the bio-ferrographic process.


2020 ◽  
Vol 72 (5) ◽  
pp. 1731-1736
Author(s):  
A.C.M. Cruz ◽  
I.L.F. Rodrigues ◽  
A.E. Souza ◽  
F.B. Knackfuss ◽  
R.L. Silveira ◽  
...  

ABSTRACT Porcine circovirus 3 (PCV-3) DNA has been detected in serum samples from apparently healthy pigs as well as pigs with different clinical conditions. Molecular detection of PCV-3 was observed in swine serum samples from Southeastern - Brazil using a nested PCR designed specifically for this study. The epidemiology and clinical aspects of PCV-3 infection were evaluated. The samples originated from 154 pigs of both genders from different production phases and with different clinical presentations, sampled from 31 pig farms visited between 2013 and 2018. In this study, PCV-3 was detected in 26.7% of samples from all populations across varying ages. Statistical association (P=0.0285) was observed only between animals with respiratory signs and PCV-3; no PCV-3-positive animal had diarrhea. No statistical association was observed between PCV-3 and age, or gender of the pigs. Because PCV-3 is a newly discovered virus, there is very little information about its epidemiology. We hope that these data can help in future studies investigating PCV-3 epidemiology.


2017 ◽  
Vol 63 (6) ◽  
pp. 1094-1100 ◽  
Author(s):  
Ivan A Katrukha ◽  
Alexander E Kogan ◽  
Alexandra V Vylegzhanina ◽  
Marina V Serebryakova ◽  
Ekaterina V Koshkina ◽  
...  

AbstractBACKGROUNDCardiac troponin T (cTnT) is an acknowledged biomarker of acute myocardial infarction (AMI) that is known to be prone to proteolytic degradation in serum. Such degradation is usually explained by the action of μ-calpain, although there could be other candidates for that role. In the current study, we explored the hypothesis that thrombin-mediated cTnT cleavage occurs as a result of the serum sample preparation.METHODScTnT degradation was studied by using immunoblotting and mass spectrometry (MS) analysis.RESULTSThe comparison of cTnT isolated from AMI heparin plasma and serum samples showed that cTnT in the plasma samples was mainly present as the full-sized molecule (approximately 35 kDa), while in serum samples it was present as a 29-kDa fragment. The incubation of recombinant cTnT, or native ternary cardiac troponin complex with thrombin or in normal human serum (NHS), resulted in the formation of a 29-kDa product that was similar to that detected in AMI serum samples. No cTnT degradation was observed when thrombin or NHS was pretreated with hirudin, a specific inhibitor of thrombin, or during incubation of troponin in normal heparin plasma. When the products of thrombin-mediated cTnT proteolysis were analyzed by MS, 2 fragments consisting of amino acid residues (aar) 2–68 and 69–288 were identified, which suggests that thrombin cleaves cTnT between R68 and S69.CONCLUSIONSThe results of this study suggest that the 29-kDa fragment of cTnT in AMI serum samples mainly appears due to the cleavage by thrombin during serum sample preparation.


Author(s):  
Mamadou Malado Jallow ◽  
Amary Fall ◽  
Serigne Fallou Wade ◽  
Ndeye Sophie Fall ◽  
Davy Kiori ◽  
...  

Herpesviruses are known to cause a diversity of clinical syndromes, ranging from minor cutaneous lesions to life-threatening illnesses, especially in immunocompromised hosts. Here, we investigate retrospectively the contribution of five human herpesviruses, including herpes simplex virus Cytomegalovirus (CMV), the Epstein–Barr virus (EBV), human herpesvirus 6, and varicella zoster virus (VZV) in serum samples collected from measles suspected patients with at least fever and rash. Sera specimens were first tested for serological evidence of measles and rubella virus infection by ELISA, and DNA extracted from an aliquot of each clinical specimen for molecular detection of human herpes viruses by RT-qPCR. A total of 3,358 specimens have been collected and tested for herpes viruses. Nearly half of the overall suspected cases were children younger than 5 years (49.4%). Of the 3,358 sera tested by ELISA, 227 (6.7%) were measles laboratory confirmed and 152 (4.5%) rubella laboratory confirmed. Herpes viruses were detected in 1763 (52.5%), and VZV was the most common with 44.3%, followed by EBV with 10.7%. Coinfections were found in 352 (20%) cases, and the most common co-detections were VZV/EBV or VZV/CMV (169 and 81 cases, respectively). A clear seasonal pattern of VZV, EBV, and CMV identification was observed, with the highest incidence between February and April each year. Results of this investigation provide more insights into cutaneous rash syndrome etiologies in patients sampled in the framework of measles/rubella surveillance in Senegal, which is useful for the guidance of both case definition revision and clinical practice as well as for public health policy.


2000 ◽  
Vol 46 (2) ◽  
pp. 273-278 ◽  
Author(s):  
Ronan Revelen ◽  
Anne Bordron ◽  
Maryvonne Dueymes ◽  
Pierre Youinou ◽  
Josiane Arvieux

Abstract Background: ELISAs with fixed endothelial cells or cell lines are widely used screening tests for anti-endothelial cell antibodies (AECAs), but spurious increases occur. We examined interferences by heteroantibodies and means to eliminate them. Methods: AECAs were measured by ELISA on fixed layers of the human endothelial cell line, EA.hy 926, in a panel of 60 patient serum samples diluted in bovine serum albumin. Heteroantibodies against fetal calf serum (FCS) proteins were demonstrated and characterized in an ELISA—the interference assay—that used FCS-coated plates and Tween 20-containing buffer as blocking agent and sample diluent, as well as by immunoblotting. Results: In 12 of 60 patient serum samples, spurious increases of AECA titers were produced by endogenous antibodies reacting with FCS proteins from culture medium that were coated onto the solid-phase at the time of cell plating. This mechanism of interference was supported experimentally by exposing extracellular matrix, varying cell density, and incubating wells with FCS alone. The heterophile antibodies were mainly IgG and IgA, and in inhibition experiments, they recognized serum proteins from goat, sheep, and horse. Washing cells free of FCS before plating, or adding FCS (100 mL/L) to the patient sample diluent eliminated spurious signals from all 30 tested sera, but the latter method had practical advantages. Conclusions: Antibodies against animal serum proteins are a frequent cause of erroneous results in cyto-ELISAs. The interference can be eliminated by simple antibody absorption in FCS-containing dilution buffer.


Diagnostics ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 51
Author(s):  
Lilla Pawlik-Sobecka ◽  
Katarzyna Sołkiewicz ◽  
Izabela Kokot ◽  
Aleksandra Kiraga ◽  
Sylwia Płaczkowska ◽  
...  

The present work aims at accessing the stability of biological material stored for diagnostic and scientific purposes. The influence of the temperature, storage time, and cyclic thawing on concentration stability of selected oxidative stress parameters in human serum was investigated. The study group consisted of 20 serum samples collected from healthy volunteers aged 18–52. The parameters whose reference ranges were not determined and to which validated determination methods did not correspond were examined by manual methods (FRAP and AOPP). Automatic methods were used to determine routine laboratory tests (albumin, total protein, bilirubin, uric acid) using the Konelab 20i® analyzer. The samples were stored at various temperatures (room temperature, 4 °C, −20 °C, −80 °C) for max 6 months and were subjected to cyclic thawing at 1 month intervals. In order to check whether any differences between the concentrations of the studied parameters existed when the samples were stored in various conditions, the paired Student t-test or Wilcoxon test and comparison to desirable bias were applied. Based on the obtained results, it was found that the temperature and time of serum sample storage significantly affected the stability of the analyzed parameters and determined different shelf lives of serum samples for oxidative stress examination. Therefore, continuing the investigation concerning the impact of storage conditions on various serum parameters seems justified due to the discrepancy between the individual results obtained by different researchers and the inconsistencies between the results of scientific research and the applicable recommendations.


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