Burnetii DNA Research Articles

High Prevalence and New Genotype of Coxiella burnetii in Ticks Infesting Camels in Somalia.

Coxiella burnetii is the causative agent of Q fever. It can infect animals, humans, and birds, as well as ticks, and it has a worldwide geographical distribution. To better understand the epidemiology of C. burnetii in Somalia, ticks infesting camels were collected from five different regions, including Bari, Nugaal, Mudug, Sool, and Sanaag, between January and March 2018. Collected ticks were tested for C. burnetii and Coxiella-like endosymbiont DNA by using IS1111, icd, and Com1-target PCR assays. Moreover, sequencing of the 16S-rRNA was conducted. Molecular characterization and typing were done by adaA-gene analysis and plasmid-type identification. Further typing was carried out by 14-marker Multi-Locus Variable-Number Tandem Repeats (MLVA/VNTR) analysis. The investigated ticks (n = 237) were identified as Hyalomma spp. (n = 227, 95.8%), Amblyomma spp. (n = 8, 3.4%), and Ripicephalus spp. (n = 2, 0.8%), and 59.1% (140/237) of them were positive for Coxiella spp. While Sanger sequencing and plasmid-type identification revealed a C. burnetii that harbours the QpRS-plasmid, MLVA/VNTR genotyping showed a new genotype which was initially named D21. In conclusion, this is the first report of C. burnetii in ticks in Somalia. The findings denote the possibility that C. burnetii is endemic in Somalia. Further epidemiological studies investigating samples from humans, animals, and ticks within the context of “One Health” are warranted.

Real-time PCR biochip for on-site detection of Coxiella burnetii in ticks

BackgroundQ fever, a zoonosis caused by Coxiella burnetii, has adverse effects on public health. Ticks are vectors of C. burnetii and they contribute to the transmission of the pathogen. A tool for rapid, sensitive, and accurate detection of C. burnetii from ticks is important for the prevention of Q fever.MethodsUltra-rapid real-time PCR (UR-qPCR) as a chip-based real-time PCR system was developed for the detection of C. burnetii from ticks. The UR-qPCR system was established and evaluated for the rapidity, sensitivity, and specificity of C. burnetii detection.ResultsC. burnetii was detected using UR-qPCR from 5644 larval, nymphal, and adult ticks from 408 pools collected from livestock and epidemiologically linked environments in two provinces, Gangwon and Jeju, in Korea. Ticks from three species were identified; Haemaphysalis longicornis accounted for the highest number, present in 333 of 408 pools (81.62%), followed by Haemaphysalis flava in 62 pools (15.19%) and Ixodes nipponensis in 13 pools (3.19%). The rapidity and sensitivity of PCR detection was demonstrated with the sufficient amplification and detection of approximately 56 copies of C. burnetii DNA with only 20 min of PCR amplification. The kappa value for the diagnostic agreement between UR-qPCR and stationary qPCR was in perfect agreement (κ = 1). PCR detection and sequencing indicated that C. burnetii was present in 5 of the 408 pools (1.23%), in which four pools contained H. longicornis and one pool contained H. flava. The infection rates of C. burnetii in the tick pools collected from Gangwon and Jeju Provinces were 1.70% and 0.58%, respectively. Phylogenetic analysis indicated a close relationship between the detected C. burnetii and those originating from goats, humans, and ticks in different countries, such as the USA, France, Germany, and Serbia.ConclusionsThe methods described in this study could be important for the prevention and control of Q fever in the two provinces. The UR-qPCR, with its features of mobility, sensitivity, and rapidity, is helpful for constructing early alert systems in the field for C. burnetii in ticks and could help alleviate the transmission of and economic damage due to Q fever.Graphical

Molecular Detection of Coxiella burnetii in Unstandardized Minas Artisanal Cheese Marketed in Southeastern Brazil

Q fever is a zoonotic disease caused by Coxiella burnetii. The causative pathogen has been detected in dairy products, which raises the possibility of consumption of dairy products as a possible route of transmission; however, this has not been adequately described. Therefore, the aim of the present study was to investigate the presence of C. burnetii in 87 samples of artisanal cheeses made from unpasteurized raw milk sold in Minas Gerais, southeastern Brazil. The cheese samples were analyzed using quantitative real-time PCR (qPCR), and the products from positive samples were sequenced. In addition, 150 people were interviewed to outline the profile of consumers and the consumption of non-inspected dairy products. The results showed that 4.6% (4/87) of the samples were positive for C. burnetii, with 99.84% identity with the reference 16S gene. Responses from consumer interviews indicate that 95.3% (143/150) of respondents consume milk and dairy products; 50% (75/150) consume fresh Minas artisanal cheese and milk at least once a day; 64.58% (93/144) do not check the inspection stamps on the packaging at the time of purchase; and 81.56% (122/146) do not know the meaning of these stamps. Thus, the presence of C. burnetii DNA in the analyzed cheeses and the consumption of unstandardized dairy products raise an alert for the risk of transmission of Q fever in the population that consumes them.

Cases of Q fever detected in residents of the novosibirsk region hospitalized with suspection of infections transmitted by ticks.

Coxiella burnetii is the causative agent of Q fever (coxiellosis), which, in addition to acute manifestations, often occurs in a latent form, is prone to chronic course and, in the absence of antibiotic therapy, has a high risk of disability or death. As a result of the presence of a wide range of clinical manifestations specific to other infectious diseases, the use of laboratory test methods (LTM) is required to make a diagnosis. The presence of Q fever anthropurgic foci in the Novosibirsk region was described in the 90s of the last century, but due attention to its laboratory diagnostics is not paid in this region. The aim of the study was to identify genetic and serological markers of the causative agent, C. burnetii, in patients of the Novosibirsk region who were admitted for treatment with fever with suspected tick-borne infections (TBIs). DNA marker of the causative agent of Q fever was detected in blood samples by real time PCR in 9 out of 325 patients. In three patients, the presence of C. burnetii DNA was confirmed by sequencing of the IS1111 and htpB gene fragments. In ELISA tests, antibodies against the causative agent of coxiellosis were detected in the blood sera of 4 patients with positive results of PCR analysis. Contact with tick was registered in 7 out of 9 patients who had C. burnetii DNA and lacked markers of other TBIs. Six people were infected in the Novosibirsk region, two suffered from tick's bite in Altai, and one case was from the Republic of Kyrgyzstan. Thus, a complex approach using both PCR analysis and ELISA provided the identification of markers of the Q fever causative agent in patients admitted with suspected TBIs, thereby differentiating it from other infections. Contact with ticks in most cases suggests that infection with C. burnetii had a transmissible pathway.

Rapid Molecular Diagnostic Sensor Based on Ball-Lensed Optical Fibers.

Given the fatal health conditions caused by emerging infectious pathogens, such as severe acute respiratory syndrome coronavirus 2, their rapid diagnosis is required for preventing secondary infections and guiding correct treatments. Although various molecular diagnostic methods based on nucleic acid amplification have been suggested as gold standards for identifying different species, these methods are not suitable for the rapid diagnosis of pathogens owing to their long result acquisition times and complexity. In this study, we developed a rapid bio-optical sensor that uses a ball-lensed optical fiber (BLOF) probe and an automatic analysis platform to precisely diagnose infectious pathogens. The BLOF probe is easy to align and has a high optical sensing sensitivity (1.5-fold) and a large detection range (1.2-fold) for an automatic optical sensing system. Automatic signal processing of up to 250 copies/reaction of DNA of Q-fever-causing Coxiella burnetii was achieved within 8 min. The clinical utility of this system was demonstrated with 18 clinical specimens (9 Q-fever and 9 other febrile disease samples) by measuring the resonant wavelength shift of positive or negative samples for Coxiella burnetii DNA. The results from the system revealed the stable and automatic optical signal measurement of DNA with 100% accuracy. We envision that this BLOF probe-based sensor would be a practical tool for the rapid, simple, and sensitive diagnosis of emerging infectious pathogens.

Prevalence and Molecular Characterization of Coxiella burnetii in Cattle, Goats, and Horses in the Republic of Korea

Coxiella burnetii is an obligate intracellular zoonotic bacterium with a global distribution. This study was conducted to investigate the prevalence of C. burnetii in different animals and to assess the potential role of these species as reservoirs of infection and transmission to humans. A total of 592 blood samples (105 beef cattle, 61 dairy cattle, 110 Korean native goats, 83 Boer goats, and 233 horses) were collected in the Republic of Korea (ROK). The C. burnetii DNA was detected from blood samples using the transposon-like repetitive region (IS1111) by PCR method. The results showed that 22.7% of the Korean-native goats, 16.4% of the dairy cattle, 15.2% of the beef cattle, 6.0% of the Boer goats, and 5.2% of the horses were positive for C. burnetii. Significant differences were found between the animal species. The univariable binary logistic regression analysis revealed that the risk of contracting C. burnetii was significantly high by 5.4-fold in Korean-native goats (95% confidence interval [CI]: 2.60%-11.27%, p = 0.000), 3.6-fold in dairy cattle (95% CI: 1.48%-8.82%, p = 0.005), and 3.3-fold in beef cattle (95% CI: 1.51%-7.28%, p = 0.003) compared with horses. A phylogenetic tree based on the IS1111 gene revealed that our sequences had 92.2%-99.9% similarity and were clustered with those detected in humans, cattle, goats, dogs, rodents, and ticks. C. burnetii circulating in the ROK exhibits genetic variation. To the best of our knowledge, this is the first study to identify C. burnetii DNA in a horse in the ROK. These results suggest that cattle, goats, and horses can be potential reservoirs for C. burnetii and play an important role in the transmission of infection. Further studies should assess the pathogenicity of C. burnetii circulating in the ROK.

Coxiella burnetii in cattle and their human contacts in a gaushala (cattle shelter) from India and its partial com1 gene sequence-based phylogenetic analysis

Q fever caused by Coxiella burnetii is an important zoonosis and has great public health significance. A total of 905 clinical samples from 387 cattle [serum (n = 387); vaginal swabs (n = 387); milk (n = 131)] and 59 serum samples from humans were collected from gaushala (cattle shelter) and screened for anti-C. burnetii IgG antibodies in the sera using an indirect-ELISA kit. Further, the samples were tested for C. burnetii DNA employing TaqMan real-time and conventional PCR assays targeting the com1 gene. In ELISA, 9.56% and 6.78% of animal and human sera samples were positive for anti-C. burnetii antibodies, respectively. Upon pathogen detection, 3.87% sera, 1.81% vaginal swabs, and 6.87% milk samples from cattle tested positive in TaqMan real-time PCR and 1.55% sera, 0.52% vaginal swabs, and 3.05% milk samples were found positive in conventional PCR. In humans, one serum sample was positive in both the PCR assays. The PCR positive samples (n = 12) were partially sequenced and the phylogenetic tree was constructed using com1 gene sequences (n = 42) from a different host and geographical areas. The study highlights infection of cattle and their human contacts in gaushala and identifies relationships between strains identified in the gaushala and those in other parts of the globe.

Serologic and molecular survey of horses to Coxiella burnetii in East of Iran a highly endemic area

There are few reports about Q fever in horse populations worldwide. This study aimed to detect the C. burnetii infection by serologic and molecular confirmation using commercial ELISA kit and real-time PCR in the East of Iran a region highly endemic. A total of 177 blood samples and 115 vaginal swabs were randomly collected from horses in East of Iran. The sera samples were analyzed for anti C.burnetii Ig G antibodies by a commercial ELISA kit and nucleic acid extraxted from vaginal samples were used to determine the C. burnetii DNA by real-time PCR assay. Antibodies were detected in 5.64 % (10/177) of sera samples and C. burnetii DNA was detected in 7.82 % (9/115) of horse vaginal samples. There was no significant difference in seroprevalence in different sex, age and breed groups. Our study showed that horses could be considered as a mild potential reservoir of C. burnetii which may be effective on horse health status. However, additional studies are needed to assess whether the horse could be considered as a relevant transmission risk indicator for Q fever.

Investigating the Role of Micromammals in the Ecology of Coxiella burnetii in Spain.

Simple SummaryCoxiella burnetii, the causal agent of human Q fever and animal Coxiellosis, is a zoonotic infectious bacterium with a complex ecology that replicates in multiple host species. However, the role of wildlife in its transmission is poorly understood. We examined 816 spleen samples obtained from ten species of micromammals and 130 vaginal swabs from Microtus arvalis females to detect the presence of C. burnetii DNA by qPCR. Our aim was assessing whether infection occurs in micromammals in Spain and what species could be relevant hosts in pathogen maintenance. The 9.7% of the spleen samples were qPCR positive. The infection prevalence level was highest (10.8%) in Microtus arvalis and also one vaginal swab was PCR positive. Positive samples were also found in Apodemus sylvaticus (8.7%), Crocidura russula (7.7%), and Rattus rattus (6.4%). A genotype II+ strain was identified in one of the positive samples from M. arvalis. The results of the study are consistent with previous findings suggesting susceptibility of micromammals to C. burnetii infection. We also provide further support to consider micromammals when tracing the origin of human Q fever cases in Europe as one of the authors probably got infected while handling M. arvalis.Coxiella burnetii, the causal agent of human Q fever and animal Coxiellosis, is a zoonotic infectious bacterium with a complex ecology that results from its ability to replicate in multiple (in)vertebrate host species. Spain notifies the highest number of Q fever cases to the ECDC annually and wildlife plays a relevant role in C. burnetii ecology in the country. However, the whole picture of C. burnetii hosts is incomplete, so this study seeks to better understand the role of micromammals in C. burnetii ecology in the country. Spleen samples from 816 micromammals of 10 species and 130 vaginal swabs from Microtus arvalis were analysed by qPCR to detect C. burnetii infection and shedding, respectively. The 9.7% of the spleen samples were qPCR positive. The highest infection prevalence (10.8%) was found in Microtus arvalis, in which C. burnetii DNA was also detected in 1 of the 130 vaginal swabs (0.8%) analysed. Positive samples were also found in Apodemus sylvaticus (8.7%), Crocidura russula (7.7%) and Rattus rattus (6.4%). Positive samples were genotyped by coupling PCR with reverse line blotting and a genotype II+ strain was identified for the first time in one of the positive samples from M. arvalis, whereas only partial results could be obtained for the rest of the samples. Acute Q fever was diagnosed in one of the researchers that participated in the study, and it was presumably linked to M. arvalis handling. The results of the study are consistent with previous findings suggesting that micromammals can be infected by C. burnetii. Our findings additionally suggest that micromammals may be potential sources to trace back the origin of human Q fever and animal Coxiellosis cases in Europe.

Vector competence of the African argasid tick Ornithodoros moubata for the Q fever agent Coxiella burnetii.

Q fever is a widespread zoonotic disease caused by the intracellular bacterium Coxiella burnetii. While transmission is primarily but not exclusively airborne, ticks are usually thought to act as vectors on the basis of early microscopy studies. However, recent observations revealed that endosymbionts of ticks have been commonly misidentified as C. burnetii, calling the importance of tick-borne transmission into question. In this study, we re-evaluated the vector competence of the African soft tick Ornithodoros moubata for an avirulent strain of C. burnetii. To this end, we used an artificial feeding system to initiate infection of ticks, specific molecular tools to monitor further infections, and culture assays in axenic and cell media to check for the viability of C. burnetii excreted by ticks. We observed typical traits associated with vector competence: The exposure to an infected blood meal resulted in viable and persistent infections in ticks, trans-stadial transmissions of infection from nymphs to adults and the ability of adult ticks to transmit infectious C. burnetii. However, in contrast to early studies, we found that infection differed substantially between tick organs. In addition, while adult female ticks were infected, we did not observe C. burnetii in eggs, suggesting that transovarial transmission is not effective. Finally, we detected only a sporadic presence of C. burnetii DNA in tick faeces, but no living bacterium was further isolated in culture assays, suggesting that excretion in faeces is not a common mode of transmission in O. moubata.

Cat fleas (Ctenocephalides felis clade ‘Sydney’) are dominant fleas on dogs and cats in New South Wales, Australia: Presence of flea-borne Rickettsia felis, Bartonella spp. but absence of Coxiella burnetii DNA

The cat flea (Ctenocephalides felis) is the most common flea species parasitising both domestic cats and dogs globally. Fleas are known vectors of zoonotic pathogens such as vector-borne Rickettsia spp. and Bartonella spp. and could theoretically transmit Coxiella burnetii, the causative agent of Q fever. A total of 107 fleas were collected from 21 cats and 14 dogs in veterinary clinics, a feline rescue organisation and a grooming salon in New South Wales, Australia, to undergo PCR detection of Bartonella spp., Rickettsia spp. and C. burnetii DNA. Morphological identification confirmed that the cat flea (C. felis) is the most common flea in New South Wales, Australia, with only a single stick fast flea, Echidnophaga gallinacea recorded. The examined fleas (n = 35) at the cox1 locus revealed five closely related C. felis haplotypes (inter-haplotype distance < 0.5%). Multiplex TaqMan qPCR targeting the gltA (Rickettsia spp.) and ssrA (Bartonella spp.) genes was positive in 22.9% (95% CI: 11.8–39.3%) and 11.4% (95% CI: 3.9–26.6%) of samples, respectively. None of the DNA isolated from fleas was positive on TaqMan qPCRs targeting the C. burnetii IS1111, Com1 and htpAB genes. Co-infection of C. felis with Bartonella henselae and Bartonella clarridgeiae was demonstrated using gltA and ssrA Illumina next-generation amplicon sequencing. These findings reinforce the importance of flea control on domestic dogs and cats to effectively control the transmission of Rickettsia felis and Bartonella spp. The flea, however, is unlikely to be a vector of C. burnetii between companion animals and humans.

Molecular detection of Coxiella burnetii infection in small mammals from Moshi Rural and Urban Districts, northern Tanzania.

Coxiella burnetii is an obligate intracellular bacterium that causes Q fever, a zoonotic disease of public health importance. In northern Tanzania, Q fever is a known cause of human febrile illness, but little is known about its distribution in animal hosts. We used a quantitative real‐time PCR (qPCR) targeting the insertion element IS1111 to determine the presence and prevalence of C. burnetii infections in small mammals trapped in 12 villages around Moshi Rural and Moshi Urban Districts, northern Tanzania. A total of 382 trapped small mammals of seven species were included in the study; Rattus rattus (n = 317), Mus musculus (n = 44), Mastomys natalensis (n = 8), Acomys wilson (n = 6), Mus minutoides (n = 3), Paraxerus flavovottis (n = 3) and Atelerix albiventris (n = 1). Overall, 12 (3.1%) of 382 (95% CI: 1.6–5.4) small mammal spleens were positive for C. burnetii DNA. Coxiella burnetii DNA was detected in five of seven of the small mammal species trapped; R. rattus (n = 7), M. musculus (n = 1), A. wilson (n = 2), P. flavovottis (n = 1) and A. albiventris (n = 1). Eleven (91.7%) of twelve (95% CI: 61.5–99.8) C. burnetii DNA positive small mammals were trapped within Moshi Urban District. These findings demonstrate that small mammals in Moshi, northern Tanzania are hosts of C. burnetii and may act as a source of C. burnetii infection to humans and other animals. This detection of C. burnetii infections in small mammals should motivate further studies into the contribution of small mammals to the transmission of C. burnetii to humans and animals in this region.

Multispacer sequence typing of Coxiella burnetii from milk and hard tick samples from ruminant farms in Lebanon.

his study was carried out to detect and characterize Coxiella burnetii in ruminant milk samples and in different tick species from seropositive farms in four Lebanese regions. Milk and tick samples were screened for C. burnetii presence by quantitative real-time PCR (qPCR) targeting IS1111 region followed by multispacer sequence typing (MST). The overall positive percentages of 9.6% (27/282) and 95.45% (84/88) for C. burnetii were recorded in ruminant milk and tick samples, respectively. In detail, the C. burnetii DNA was recorded in 52/54 (96.3%) of Rhipicephalus annulatus, 20/21 (95.24%) of Rhipicephalus turanicus, 6/6 (100%) of Hyalomma anatolicum, 5/6 (83.3%) of Rhipicephalus sanguineus and 1/1 of Rhipicephalus bursa. After genotyping of some IS1111-positive samples (17/111), different MST genotypes were identified. Out of 15 positive ticks, 10 were infected with MST2 genotype, 4 were infected with MST7 genotype and 1 was infected with MST57. Moreover, genotypes MST20 and MST58 were found in one cow and one goat milk samples, respectively. The present study confirmed the high genetic diversity of C. burnetii in Lebanon.

Coxiella burnetii in slaughterhouses in Brazil: A public health concern.

Q fever is an important zoonosis, yet it is often neglected and can present large outbreaks, as observed in the Netherlands. In the past few years, cases of Q fever have been described in Brazil; however, the epidemiological situation of Q fever in ruminants, the main reservoir of the pathogen, is unknown in this country. Our study aimed to estimate the prevalence of C. burnetii in cattle sent to slaughterhouses using an immunofluorescence assay (IFA) and quantitative real-time PCR (qPCR). From 1515 cattle serum samples collected from nine slaughterhouses, 23.8% (360/1515) were serologically positive by IFA (cutoff titer>1:64), indicating past or recent exposure to C. burnetii infection. Among the 54 cities sampled during the study, 83.3% (45/54) had at least one seropositive animal. Subsequently, all seropositive samples were submitted to qPCR for C. burnetii DNA, and 12.2% (44/360) of the sera were qPCR positive, which indicates bacteremia and suggests active or recent infection. The results highlight the risk for abattoir workers that results from exposure to contaminated aerosols produced during slaughter procedures. Moreover, the heat maps that were construction from the positive samples demonstrate the widespread distribution of C. burnetii in the State of São Paulo, Brazil and denotes the need for surveillance and preventive measures to reduce the prevalence in cattle.

Molecular Detection of Microorganisms Associated with Small Mammals and Their Ectoparasites in Mali.

Small mammals are the natural reservoirs for many zoonotic pathogens. Using molecular tools, we assessed the prevalence of bacteria and protozoans in small mammals and their ectoparasites in Faladjè, Bougouni, and Bamoko, Mali. A total of 130 small mammals belonging to 10 different species were captured, of which 74 (56.9%) were infested by ectoparasites, including Laelaps echidnina, Xenopsylla cheopis, Amblyomma variegatum, Rhipicephalus sanguineus sensu lato, and Haemaphysalis spp. nymphs. DNA of Bartonella was found in 14/75 (18.7%), 6/48 (12.5%), and 3/7 (42.8%) small mammals from Faladjè, Bougouni, and Bamako, respectively. In Faladjè, Bartonella DNA was detected in 31/68 (45.6%) of L. echidnina and 14/22 (63.6%) of X. cheopis. In Bougouni, it was found in 2/26 (7.7%) of L. echidnina and 10/42 (23.8%) of X. cheopis. The sequences of Bartonella obtained from small mammals were close to those of Bartonella mastomydis, Bartonella elizabethae, and uncultured Bartonella spp. In Faladjè, Coxiella burnetii DNA was detected in 64.4% (29/45) of Haemaphysalis spp. ticks, 4.5% (2/44) of Mastomys erythroleucus, 12.5% (1/8) of Praomys daltoni, and 1.5% (1/68) of L. echidnina. We found DNA of Wolbachia in X. cheopis from Faladjè and DNA of Rickettsia africae and Ehrlichia ruminantium in Am. variegatum from Bougouni. The results of our study show that several small mammal species harbor and may serve as potential reservoirs of Bartonella spp., likely to play a major role in the maintenance, circulation, and potential transmission of bacteria in Mali. The pathogenicity of these bacteria for humans or animals remains to be demonstrated.

Seroprevalence and Molecular Characterization of Coxiella burnetii in Cattle in the Republic of Korea.

This study was conducted to determine the prevalence of Coxiella burnetii in cattle and how that prevalence is influenced by cattle breed and growth type. A total of 491 cattle [cattle breed: 216 dairy cattle and 275 beef cattle; growth type: indoor housed (n = 294) and grazing (n = 197)] were used. The presence of C. burnetii DNA and antibodies was detected from blood and serum samples using polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. The overall prevalence of C. burnetii was: 10.8% (95% CI: 8.0–13.5%) using PCR and 8.8% (95% CI: 6.3–11.3%) using ELISA. The prevalence of C. burnetii was significantly higher in beef cattle than in dairy cattle using both PCR (13.5% vs. 7.4%; P = 0.032) and ELISA (14.5% vs. 1.4%; P = 0.000), respectively. Comparison by growth type revealed that C. burnetii infection was significantly higher in grazing cattle than in housed cattle when using both PCR (24.9% vs. 1.4%; P = 0.000) and ELISA (21.3% vs. 0.3%; P = 0.000). Beef cattle were at a significantly higher risk of contracting C. burnetii compared with dairy cattle (odds ratio = 3.20, 95% CI: 1.80–5.67; P = 0.000). The risk of contracting C. burnetii in grazing cattle was increased by 32.57-fold (95% CI: 12.84–82.61; P = 0.000) compared with indoor housed cattle. The phylogenetic analysis based on the IS1111 gene revealed that our sequences grouped with human, tick, goat, and cattle isolates/strains found in several countries. C. burnetii sequences circulating in the Republic of Korea exhibit genetic variations. Thus, grazing is a high risk factor for the prevalence and transmission of C. burnetii.

Transstadial Transmission from Nymph to Adult of Coxiella burnetii by Naturally Infected Hyalomma lusitanicum.

Coxiella burnetii (Derrick) Philip, the causative agent of Q fever, is mainly transmitted by aerosols, but ticks can also be a source of infection. Transstadial and transovarial transmission of C. burnetii by Hyalomma lusitanicum (Koch) has been suggested. There is a close relationship between this tick species, wild animals and C. burnetii but the transmission in a natural environment has not been demonstrated. In this study, we collected 80 engorged nymphs of H. lusitanicum from red deer and wild rabbits. They moult to adults under laboratory conditions and we feed them artificially through silicone membranes after a preconditioning period. C. burnetii DNA was tested in ticks, blood and faeces samples using real-time PCR. The pathogen was found in 36.2% of fed adults, demonstrating that transstadial transmission from nymph to adult occurs in nature. The presence of DNA in the 60.0% of blood samples after artificial feeding confirms that adults transmit the bacteria during feeding. Further studies are needed about co-feeding and other possible transmission routes to define the role of this tick species in the cycle of C. burnetii.

Molecular investigation, isolation and phylogenetic analsysis of Coxiella burnetii from aborted fetus and ticks

Q fever is a zoonotic infection threatening human health, causing abortions in cattle, sheep and goats. Coxiella burnetii (C. burnetii) also causes serious problems such as low birth weight, infertility. This study is the first exemplary for analysis of Q fever around Black Sea region in Turkey. In the study, a total of 270 aborted fetuses (171 cattle, 79 sheep, 20 goats) and 1069 tick samples were aimed to be searched by PCR method. C. burnetii DNA was detected in 8 (2.96 %) of 270 sheep specimens while it could not be found in cattle and goat specimens. 406 sample pools were created from 1069 tick samples (490 male, 579 female) collected from 254 farm animals (187 cattle, 54 sheep, 13 goats) and 11 of these were stated positive. Tick species determined as C. burnetii positive were Hyalomma marginatum, Hyalomma anoliticum excavatum, Hyalomma detritum and Boophilus annulatus. Agent isolation was carried out within embryonated eggs. Agents were stained with Giemsa and was showed. Sequence analysis was performed for TUR/SAM/coxiella_1 (MN917207) isolate and phylogenetic tree was created. This tree, created in compliance with IS1111 transposon gene, did not form different branches in regard to host affiliation (goat, sheep, tick, human) and geographical distribution. As a result, an important zoonotic agent, C. burnetii was diagnosed in sheep aborted fetuses and the infection was proved to have spread among sheep herds in Black Sea region. Besides, 4 separate tick species found in our region hosted the agent and were found important for infection.

Selected Endemic Zoonoses in Pigs Presenting for Slaughter in Kampala, Uganda.

.Leptospirosis, brucellosis, and Q fever (coxiellosis) are bacterial zoonoses that cause acute febrile illness in people as well as reproductive losses in pigs. Pig keeping is an increasingly important livelihood to millions of smallholder farmers in Uganda because of exponential increases in demand for pork. The prevalence of leptospirosis and Q fever in pigs is unknown, and the few studies of porcine brucellosis have estimated a range of seroprevalence. Therefore, we undertook a prevalence survey of leptospirosis, brucellosis, and Q fever in pigs using quantitative real-time PCR to determine the potential importance of these zoonoses to the growing pig sector in Uganda. Six hundred forty-nine pigs were sampled in 2015–2016 at an urban pork slaughterhouse. Ten percent of pigs (n = 68) had leptospiral DNA in either their kidney or reproductive tissue. In adjusted analyses, variables predictive of leptospiral status included female sex (odds ratio [OR]: 2.37, P < 0.01) and pigs sampled in March 2016 (OR: 2.23, P = 0.02) and October 2016 (OR: 0.30, P = 0.04). DNA fingerprinting revealed circulation of at least four distinct serovars in these pigs. Brucella spp. and Coxiella burnetii DNA were not detected in any sampled pig. This is the first report of widespread circulation of pathogenic Leptospira spp. in pigs in Uganda, suggesting that leptospirosis likely has a greater impact on the health of pigs than was previously recognized. Pig farmers, pig traders, and slaughterhouse workers may be at greatest occupational risk because of their direct contact with infective leptospires in aborted fetuses, bodily fluids, and other tissues.

Comparison of Coxiella burnetii Excretion between Sheep and Goats Naturally Infected with One Cattle-Associated Genotype.

The main reservoir of Coxiella (C.) burnetii are ruminants. They shed the pathogen through birth products, vaginal mucus, faeces and milk. A direct comparison of C. burnetii excretions between naturally infected sheep and goats was performed on the same farm to investigate species-specific differences. The animals were vaccinated with an inactivated C. burnetii phase I vaccine at the beginning of the study period for public health reasons. Vaginal and rectal swabs along with milk specimens were taken monthly during the lambing period and once again at the next lambing season. To estimate the environmental contamination of the animals’ housings, nasal swabs from every animal were taken simultaneously. Moreover, dust samples from the windowsills and straw beddings were collected. All samples were examined by qPCR targeting the IS1111 gene and the MLVA/VNTR typing method was performed. Whole genome sequencing was applied to determine the number of IS1111 copies followed by a calculation of C. burnetii genome equivalents of each sample. The cattle-associated genotype C7 was detected containing 29 IS1111 copies. Overall, goats seem to shed more C. burnetii through vaginal mucus and in particular shed more and for longer via the rectal route than sheep. This is supported by the larger quantities of C. burnetii DNA detected in caprine nasal swabs and environmental samples compared to the ovine ones. Transmission of C. burnetii from cattle to small ruminants must also be considered.