scholarly journals Detection of Coxiella burnetii (Query Fever) DNA by Nested-PCR in Beef Cattle from Ampel Slaughterhouse, Boyolali Regency, Middle Java, Indonesia

2021 ◽  
Vol 11 (2) ◽  
pp. 267-272
Author(s):  
Eko Prasetyo Nugroho ◽  
Agus Setiyono ◽  
Upik Kesumawati Hadi ◽  
Wiwin Winarsih ◽  
Dwi Astutid
Keyword(s):  
Beef Cattle ◽  
Coxiella Burnetii ◽  
Nested Pcr ◽  
Q Fever ◽  
Local Breed ◽  
Obligate Intracellular ◽  
Positive Individual ◽  
Dna Fragment ◽  
Simmental Cattle ◽  
Lung And Kidney

Coxiella burnetii (C. burnetii) is a Gram-negative and obligate intracellular bacterium that causes Query fever (Q fever). The aim of the present study was to detect C. burnetii in beef cattle from Ampel slaughterhouse at Boyolali Regency, Middle Java, Indonesia. Spleen, heart, liver, lung, and kidney samples were collected from 100 cattle and used for Nested-PCR (nPCR) with four types of primers (OMP1, OMP2, OMP3, and OMP4). Five stages of pooling extraction were performed on 100 individual samples. The nPCR amplified a 437 bp DNA fragment from the fifth pool on the sampled heart, lung, and spleen. Furthermore, 10 individual samples from the fifth pool were re-tested by nPCR to find out the number of positive individual samples. Of 10 samples, the obtained result indicated the presence of C. burnetii DNA in 7 samples, 6 from Simmental cattle and 1 from Ongole cattle. Therefore, it can be strongly suspected that there are 7 out of 100 local breed beef cattle positive of Q fever at Boyolali Regency, Middle Java, Indonesia.

scholarly journals Immunohistochemical Detection of Coxiella burnetii in Cattle Spleen Organs from Ampel Slaughterhouse, Boyolali Regency

2021 ◽  
Vol 4 (1) ◽  
pp. 48
Author(s):  
Eko Prasetyo Nugroho ◽  
Agus Setiyono ◽  
Upik Kesumawati Hadi ◽  
Wiwin Winarsih ◽  
Dwi Astuti
Keyword(s):  
Coxiella Burnetii ◽  
Q Fever ◽  
Intracellular Bacteria ◽  
Bacterial Disease ◽  
Immunohistochemical Detection ◽  
Immunohistochemical Examination ◽  
Obligate Intracellular ◽  
Positive Individual ◽  
Liver And Kidney ◽  
Obligate Intracellular Bacteria

Q-fever is a zoonotic bacterial disease that caused by Coxiella burnetii. These microorganism are gram negative and obligate intracellular bacteria. This study was conducted to detect C. burnetii in cattle organs which collected from Ampel slaughterhouse Boyolali Regency. In this study, spleen, heart, lung, liver and kidney were collected from 100 cattle. The samples were tested by immunohistochemical (IHC) method using polyclonal anti- C. burnetii antibodies. Immunohistochemical examination found the presence of C. burnetii in the cytoplasm of macrophage cells with specific brown color only in the spleen as many as 4 out of 100 cattle showing immunoreactive (4%). The four positive individual samples were from Simental cattle. These results indicate that Q-fever was found in local cattle in Boyolali Regency.

A study of the correlation between Coxiella burnetii seropositivity and abortions in sheep in Eastern and Southeastern Turkey

2016 ◽  
Author(s):  
Ayse Kilic ◽  
Hakan Kalender
Keyword(s):  
Immunosorbent Assay ◽  
Coxiella Burnetii ◽  
Q Fever ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Eastern Anatolia ◽  
Obligate Intracellular Bacterium ◽  
Serum Samples ◽  
Obligate Intracellular ◽  
Elisa Kit

Q fever is a zoonotic disease that occurs worldwide and is caused by the obligate intracellular bacterium Coxiella burnetii. Infected animals are usually asymptomatic, but infection can cause abortion and stillbirth in ruminants. The main purpose of this study was to evaluate prevalance of Coxiella burnetii infection in aborted and nonaborted sheep serum samples in Eastern Anatolia region by using enzyme-linked immunosorbent assay (ELISA). The determine of prevalance in sheep flocks from four provinces (Elazig, Malatya, Tunceli, Bitlis) and tested for anti-C.burnetii antibody detection, by means of Chekit Q fever Elisa kit. 350 serum samples obtained from flocks belonging aborted sheep showed that a total of 56 (16%) were detected seropositivity, whereas 171 serum samples obtained from nonaborted sheep flocks in 13 of the 171 (7.60%) for C.burnetii in seropositivity were observed. Coxiellosis should be considered an important cause of sheep with abortion history and nonaborted in Elazig and neighboring provinces.

scholarly journals Serological characterization of surface-exposed proteins of Coxiella burnetii

Microbiology ◽  
2014 ◽  
Vol 160 (12) ◽  
pp. 2718-2731 ◽  
Author(s):  
Jun Jiao ◽  
Xiaolu Xiong ◽  
Yong Qi ◽  
Wenping Gong ◽  
Changsong Duan ◽  
...  
Keyword(s):  
Coxiella Burnetii ◽  
Bioinformatics Analysis ◽  
Q Fever ◽  
Obligate Intracellular ◽  
Rickettsia Rickettsii ◽  
Serological Characterization ◽  
Vaccine Antigens ◽  
Mycoplasma Pneumonia ◽  
Dimensional Electrophoresis

The obligate intracellular Gram-negative bacterium Coxiella burnetii causes Q fever, a worldwide zoonosis. Here we labelled Cox . burnetii with biotin and used biotin-streptavidin affinity chromatography to isolate surface-exposed proteins (SEPs). Using two-dimensional electrophoresis combined with mass spectrometry, we identified 37 proteins through bioinformatics analysis. Thirty SEPs expressed in Escherichia coli (recombinant SEPs, rSEPs) were used to generate microarrays, which were probed with sera from mice experimentally infected with Cox. burnetii or sera from Q fever patients. Thirteen rSEPs were recognized as seroreactive, and the majority reacted with at least 50 % of the sera from mice infected with Cox. burnetii but not with sera from mice infected with Rickettsia rickettsii, R. heilongjiangensis, or R. typhi. Further, 13 proteins that reacted with sera from patients with Q fever did not react with sera from patients with brucellosis or mycoplasma pneumonia. Our results suggest that these seroreactive SEPs have potential as serodiagnostic antigens or as subunit vaccine antigens against Q fever.

scholarly journals Q fever

2012 ◽  
Vol 33 (4) ◽  
pp. 170
Author(s):  
Robert Norton
Keyword(s):  
Coxiella Burnetii ◽  
Q Fever ◽  
Environmental Sampling ◽  
High Likelihood ◽  
Obligate Intracellular Bacterium ◽  
Wet Season ◽  
Obligate Intracellular ◽  
Chronic Q Fever ◽  
Specific Symptoms ◽  
Occupational Contact

Q fever is a zoonosis caused by the obligate intracellular bacterium Coxiella burnetii. North Queensland has some of the highest rates of Q fever notifications in Australia. The clinical diagnosis of Q fever can be difficult with non-specific symptoms. Up to 5% of cases will develop chronic Q fever with a high likelihood of endocarditis. Diagnosis is essentially by serology. In North Queensland cases have clustered in relatively new, semi-rural suburbs which lie adjacent to native bushland. Native mammals are attracted to new growth in these cleared areas, particularly after the wet season. There is little or no occupational contact with traditional sources of Q fever such as cattle. Seroprevalence studies on native mammals have shown higher levels of seropositivity in native mammals than in cattle. It is postulated that the increase in human cases seen from these areas are a direct effect of interaction between native mammals and humans. Further studies on environmental sampling is currently under way.

scholarly journals Direct Identification of Coxiella burnetii Plasmids in Human Sera by Nested PCR

1998 ◽  
Vol 36 (8) ◽  
pp. 2210-2213 ◽  
Author(s):  
G. Q. Zhang ◽  
A. Hotta ◽  
M. Mizutani ◽  
T. Ho ◽  
T. Yamaguchi ◽  
...  
Keyword(s):  
Coxiella Burnetii ◽  
Nested Pcr ◽  
Q Fever ◽  
Serum Samples ◽  
Specific Primers ◽  
Direct Identification ◽  
Human Sera ◽  
Nested Pcr Assays ◽  
Acute Phase Serum ◽  
Specific Sequences

Nested PCR assays were used for the direct identification ofCoxiella burnetii plasmids in human sera. A total of 81 serum samples from 81 patients with Q fever were tested by nested PCR with four sets of primers. The first set of primers was used to detect the genomic sequences. The second set of primers was used to detect the conserved sequences of the plasmids. Another two sets of primers were used to identify the QpH1 and QpRS plasmids. QpH1 and QpRS plasmid-specific sequences were identified in 40 (49.4%) and 24 (29.6%) of the serum samples, respectively. Both of the QpH1 and QpRS plasmid-specific sequences were detected in 5 (8.6%) of the serum samples but were not found in 12 (20.7%) of the serum samples. Furthermore, all of the 23 acute-phase serum samples were positive for the QpH1 plasmid and negative for the QpRS plasmid. Nested PCR with plasmid-specific primers appears to be a useful method for the direct typing of C. burnetii plasmids in human sera.

scholarly journals Coxiella burnetii Inhibits Activation of Host Cell Apoptosis through a Mechanism That Involves Preventing Cytochrome c Release from Mitochondria

2007 ◽  
Vol 75 (11) ◽  
pp. 5282-5289 ◽  
Author(s):  
Anja Lührmann ◽  
Craig R. Roy
Keyword(s):  
Cell Death ◽  
Host Cell ◽  
Coxiella Burnetii ◽  
Mammalian Cells ◽  
Q Fever ◽  
Host Cells ◽  
Intracellular Pathogen ◽  
Obligate Intracellular ◽  
Cell Death Pathway ◽  
Obligate Intracellular Pathogen

ABSTRACT Coxiella burnetii is an obligate intracellular pathogen and the etiological agent of the human disease Q fever. C. burnetii infects mammalian cells and then remodels the membrane-bound compartment in which it resides into a unique lysosome-derived organelle that supports bacterial multiplication. To gain insight into the mechanisms by which C. burnetii is able to multiply intracellularly, we examined the ability of host cells to respond to signals that normally induce apoptosis. Our data show that mammalian cells infected with C. burnetii are resistant to apoptosis induced by staurosporine and UV light. C. burnetii infection prevented caspase 3/7 activation and limited fragmentation of the host cell nucleus in response to agonists that induce apoptosis. Inhibition of bacterial protein synthesis reduced the antiapoptotic effect that C. burnetii exerted on infected host cells. Inhibition of apoptosis in C. burnetii-infected cells did not correlate with the degradation of proapoptotic BH3-only proteins involved in activation of the intrinsic cell death pathway; however, cytochrome c release from mitochondria was diminished in cells infected with C. burnetii upon induction of apoptosis. These data indicate that C. burnetii can interfere with the intrinsic cell death pathway during infection by producing proteins that either directly or indirectly prevent release of cytochrome c from mitochondria. It is likely that inhibition of apoptosis by C. burnetii represents an important virulence property that allows this obligate intracellular pathogen to maintain host cell viability despite inducing stress that would normally activate the intrinsic death pathway.

scholarly journals Phagocytosis of Apoptotic Cells Increases the Susceptibility of Macrophages to Infection with Coxiella burnetii Phase II through Down-Modulation of Nitric Oxide Production

2004 ◽  
Vol 72 (4) ◽  
pp. 2075-2080 ◽  
Author(s):  
Dario S. Zamboni ◽  
Michel Rabinovitch
Keyword(s):  
Nitric Oxide ◽  
Coxiella Burnetii ◽  
Q Fever ◽  
Bacterial Load ◽  
Peritoneal Macrophages ◽  
No Synthase ◽  
Obligate Intracellular Bacterium ◽  
Biologically Relevant ◽  
Obligate Intracellular ◽  
Synthase Inhibitor

ABSTRACT Coxiella burnetii, the agent of Q fever in humans and coxiellosis in other mammals, is an obligate intracellular bacterium which is sheltered and multiplies within typically large phagolysosome-like replicative vacuoles (LRVs). We have previously shown that, compared with fibroblasts, mouse resident peritoneal macrophages control the development of LRVs and bacterial multiplication within these vacuoles. Earlier experiments with the nitric oxide (NO) synthase inhibitor aminoguanidine (AG) revealed that the control is exerted by NO induced by the bacteria. We report here that phagocytosis of apoptotic-like, but not of aldehyde-killed, lymphocytes by the macrophages reduced the production of NO induced by the bacteria and increased the development of LRVs and, therefore, the total bacterial load in the cultures. Experiments with macrophages from mice deficient for inducible NO synthase (iNOS−/−) confirmed the involvement of NO in the control of infection, since neither apoptotic lymphocytes nor AG affected the development of LRVs in these phagocytes. Since macrophages are important for the clearance of apoptotic bodies and C. burnetii is able to induce apoptosis in human monocytes, the phenomenon shown here may be biologically relevant to the development of Q fever and coxiellosis.

scholarly journals Coxiella burnetii in Infertile Dairy Cattle With Chronic Endometritis

2018 ◽  
Vol 55 (4) ◽  
pp. 539-542 ◽  
Author(s):  
Davide De Biase ◽  
Alessandro Costagliola ◽  
Fabio Del Piero ◽  
Rossella Di Palo ◽  
Domenico Coronati ◽  
...  
Keyword(s):  
Dairy Cattle ◽  
Coxiella Burnetii ◽  
Animal Species ◽  
Bacterial Culture ◽  
Q Fever ◽  
Premature Delivery ◽  
Chronic Endometritis ◽  
Obligate Intracellular ◽  
Obligate Intracellular Pathogen ◽  
First Time

Coxiella burnetii is an obligate intracellular pathogen and the cause of Q fever in many animal species and humans. Several studies have reported the association between C. burnetii and abortion, premature delivery, stillbirth, and weak offspring. However, no solid evidence indicates that C. burnetii causes endometritis, subfertility, and retained fetal membranes. For this study, histopathological and PCR evaluation were performed on 40 uterine biopsies from dairy cattle with poor fertility. Uterine swabs were concurrently tested with microbiology assays. The endometrial biopsies of 30 cows did not have any significant lesions, and no pathogens were identified by aerobic bacterial culture and PCR. Ten cows were PCR-positive for C. burnetii and negative for other pathogens by aerobic bacterial culture and PCR. These 10 cases revealed a mild to severe chronic endometritis admixed with perivascular and periglandular fibrosis. Immunohistochemical evaluation of C. burnetii PCR-positive biopsies identified, for the first time, the presence of intralesional and intracytoplasmic C. burnetii in macrophages in the endometrium of cattle.

scholarly journals Q Fever in the Greek Island of Crete: Detection, Isolation, and Molecular Identification of Eight Strains of Coxiella burnetii from Clinical Samples

1998 ◽  
Vol 36 (7) ◽  
pp. 2063-2067 ◽  
Author(s):  
Ioanna Spyridaki ◽  
Achilleas Gikas ◽  
Diamantis Kofteridis ◽  
Anna Psaroulaki ◽  
Yannis Tselentis
Keyword(s):  
Coxiella Burnetii ◽  
Nested Pcr ◽  
Q Fever ◽  
Buffy Coat ◽  
Clinical Samples ◽  
Polymorphism Analysis ◽  
Antibody Technique ◽  
Presenting Symptoms ◽  
Specificity And Sensitivity ◽  
Reference Strains

Over a period of 6 years (1989 to 1995), serum samples from 3,300 patients suspected to be infected by Coxiella burnetii were assayed for the presence of antibodies against antigen phase II of the microorganism by the indirect immunofluorescence antibody technique (IFAT). One hundred fifty-two cases were recorded, and blood samples from 17 patients were cultured for the isolation of the pathogen. By a centrifugation shell vial technique, eight strains were isolated from patients suffering from acute Q fever. The microorganism was detected in the cultures by IFAT, by Gimenez staining, and by the cytopathogenic effect on Vero and human embryonic lung (HEL) cells. PCR followed by restriction fragment length polymorphism analysis was used to confirm the diagnosis and identify the Coxiella burnetii strains within the cell cultures as well as to compare them with reference strains. In order to avoid time-consuming cultures, to achieve direct detection of Coxiella burnetii in clinical samples (blood, buffy coat, etc.), and to increase the specificity and sensitivity of the detection, nested PCR was performed. The first step of DNA extraction was performed with the QIAamp blood kit 250. For the second step of the PCR assays, the conditions of temperature and times of recycling were properly modified, and the microorganism was detected within 4 h. Our study demonstrates that Q fever is an endemic disease in Crete and that the diagnosis of Coxiella burnetii infection can be rapidly achieved by the detection of the microorganism in buffy coat samples by nested PCR. Although the presenting symptoms of the disease in this study differed from those in other studies, the Cretan strains do not differ genotypically from the reference strains (Nine Mile and Q212).

scholarly journals Presence of Coxiella burnetii DNA in the Environment of the United States, 2006 to 2008

2010 ◽  
Vol 76 (13) ◽  
pp. 4469-4475 ◽  
Author(s):  
Gilbert J. Kersh ◽  
Teresa M. Wolfe ◽  
Kelly A. Fitzpatrick ◽  
Amanda J. Candee ◽  
Lindsay D. Oliver ◽  
...  
Keyword(s):  
United States ◽  
Coxiella Burnetii ◽  
Q Fever ◽  
The United States ◽  
Obligate Intracellular ◽  
The Past ◽  
Background Levels ◽  
Post Offices ◽  
Animal Populations ◽  
Domestic Livestock

ABSTRACT Coxiella burnetii is an obligate intracellular bacterium that causes the zoonotic disease Q fever. Because C. burnetii is highly infectious, can survive under a variety of environmental conditions, and has been weaponized in the past, it is classified as a select agent and is considered a potential bioweapon. The agent is known to be present in domestic livestock and in wild animal populations, but the background levels of C. burnetii in the environment have not been reported. To better understand the amount of C. burnetii present in the environment of the United States, more than 1,600 environmental samples were collected from six geographically diverse parts of the United States in the years 2006 to 2008. DNA was purified from these samples, and the presence of C. burnetii DNA was evaluated by quantitative PCR of the IS1111 repetitive element. Overall, 23.8% of the samples were positive for C. burnetii DNA. The prevalence in the different states ranged from 6 to 44%. C. burnetii DNA was detected in locations with livestock and also in locations with primarily human activity (post offices, stores, schools, etc.). This study demonstrates that C. burnetii is fairly common in the environment in the United States, and any analysis of C. burnetii after a suspected intentional release should be interpreted in light of these background levels. It also suggests that human exposure to C. burnetii may be more common than what is suggested by the number of reported cases of Q fever.

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