Brucella microti‐like prevalence in French farms producing frogs

In the last 10 years, many atypical novel members of Brucella species have been reported, including several Brucella inopinata‐like strains in wild‐caught and “exotic” amphibians from various continents. In 2017, a strain of Brucella was isolated for the first time in animals from a French farm producing frogs—Pelophylax ridibundus—for human consumption and identified as B. microti‐like. Following this first isolation, investigations were performed in this farm as well as in the farm of the research unit that provided the domestic frog strain to estimate the prevalence of B. microti‐like infection and its presence in the surrounding environment. Farming practices were investigated and samples including frogs at different development stages, surface tank swabs, water, feed and soil were analysed by real‐time PCR and bacteriological methods. High B. microti‐like prevalence values (higher than 90%) were obtained in frog samples in the commercial farm, and its presence was highlighted in the environmental samples except feed. In the research unit farm, B. microti‐like species was also isolated and detected in frog and environmental samples. These results show that B. microti‐like organisms are able to colonize amphibians and persist in their environment. Its presence could constitute a possible risk for consumers and workers proving the importance of assessing the zoonotic and pathogenic potentials of these new and atypical Brucella species.     

The Prevalence of Brucellosis in Cattle,Goats and Humans in Rural Uganda: A Comparative Study

A cross‐sectional study was conducted to determine the presence of brucellosis in cattle, goats and humans in farms from south‐western Uganda and identify risk factors associated with brucellosis in these three host groups. Data and serum samples were collected from 768 cattle, 315 goats and 236 humans, with 635 samples of bovine milk, from 70 farms in two different study areas in south‐western Uganda. Sera from livestock were tested with the Rose Bengal Plate test, using B. abortus and B. melitensis antigens, and human sera were tested with a commercial IgG/IgM lateral flow assay. Milk samples were tested using the OIE‐approved milk ring test. Screening tests for brucellosis were positive in 14% of cattle serum, 29% of bovine milk, 17% of goat serum and 11% of human serum samples. There were significant differences in the test prevalence of brucellosis by study site, with levels higher in the study area near Lake Mburo National Park than in the study area near Queen Elizabeth National Park. Multivariable regression models identified risk factors associated with increasing test positivity at the individual and farm levels for cattle, goats and humans. Positive associations were seen between increasing seropositivity of brucellosis in goats, cattle and humans. Results of multivariable analyses suggest that improvements in farm biosecurity and hygiene may reduce the risk of brucellosis on the farm and suggest a role for ticks in bovine brucellosis. Although cattle are the focus of brucellosis control in Uganda, the significant associations between seropositivity in humans and seropositivity in goats suggest that brucellosis in goats may be an important contributor to the epidemiology of the disease on the farm.     

GFP tagging of Brucella melitensis Rev1 allows the identification of vaccinated sheep

Brucellosis is a worldwide zoonosis causing important economic loss and a public health problem. Small ruminants are the preferred hosts of Brucella melitensis and thus the main source of human infections. Effective control of sheep and goat brucellosis has been achieved in several countries through vaccination with the live‐attenuated B. melitensis Rev1 vaccine. However, Rev1 induces a long‐lasting serological response that hinders the differentiation between infected and vaccinated animals. A Rev1::gfp strain expressing constitutively the Green Fluorescent Protein (GFP) was built by stable insertion of a mini‐Tn7‐gfp in the glmS‐recG non‐codifying chromosomal region. An associated indirect ELISA‐GFP was developed to identify anti‐GFP antibodies in vaccinated animals. The resulting Rev1::gfp kept the biological properties of the Rev1 reference strain, including residual virulence and efficacy in mice, and was readily distinguished from Rev1 and other Brucella field strains by direct visualization under ultraviolet illumination, fluorescence microscopy and a multiplex PCR‐GFP. The Rev1::gfp strain did not elicit anti‐GFP antibodies itself in lambs but when applied in combination with recombinant GFP induced an intense and long‐lasting (>9 months) anti‐GFP serological response readily detectable by the ELISA‐GFP. Overall, our results confirm that Rev1 GFP‐tagging can be a suitable alternative for identifying vaccinated sheep in infected contexts.     

Distinction Between Persistent and Transient Infection in a Bovine Viral Diarrhoea (BVD) Control Programme: Appropriate Interpretation of Real‐Time RT‐PCR and Antigen‐ELISA Test Results

Control of bovine viral diarrhoea (BVD) in Belgium is currently implemented on a voluntary basis at herd level and mainly relies on detection and culling of persistently infected (PI) animals. The present field study was conducted during the winter of 2010/2011 to assess the performances of diagnostic assays used in the testing scheme for BVD as proposed by the two Belgian regional laboratories. Individual blood samples were collected from 4972 animals, and individual samples from the same herd were pooled (maximum of 30 individual samples per pool) and screened for the presence of Bovine Viral Diarrhoea Virus (BVDV)‐specific RNA using a commercial real‐time RT‐PCR test (ADIAGENE). Individual samples from positive pools were then tested in parallel with the same RT‐PCR test and with an antigen‐capture ELISA test (IDEXX) to detect viremic animals. This study demonstrated that individual results differed according to the type of assay used (P < 0.001): 140 animals (2.8%) were positive by RT‐PCR and 72 (1.4%) by antigen‐ELISA. A second blood sample was taken 40 days later from 74 PCR positive animals to detect persistent viremia: 17 (23%) of these were still PCR positive and considered to be PI and the 57 that no longer tested positive were assumed to be transiently infected (TI) animals. All PI animals were positive also by antigen‐ELISA at both time points. Among TI animals, 10 (16%) were positive by antigen‐ELISA at the first but none at the second sampling. A highly significant difference in cycle threshold (C_t) values obtained by RT‐PCR was observed between PI and TI animals. ROC analysis was performed to establish thresholds to confirm with high probability that an animal is PI, based on the result of RT‐PCR test performed on a single individual blood sample.     

High Prevalence of Anaplasma spp. in Small Ruminants in Morocco

The prevalence of infection by Anaplasma spp. (including Anaplasma phagocytophilum) was determined using blood smear microscopy and PCR through screening of small ruminant blood samples collected from seven regions of Morocco. Co‐infections of Anaplasma spp., Babesia spp, Theileria spp. and Mycoplasma spp. were investigated and risk factors for Anaplasma spp. infection assessed. A total of 422 small ruminant blood samples were randomly collected from 70 flocks. Individual animal (breed, age, tick burden and previous treatment) and flock data (GPS coordinate of farm, size of flock and livestock production system) were collected. Upon examination of blood smears, 375 blood samples (88.9%) were found to contain Anaplasma‐like erythrocytic inclusion bodies. Upon screening with a large spectrum PCR targeting the Anaplasma 16S rRNA region, 303 (71%) samples were found to be positive. All 303 samples screened with the A. phagocytophilum‐specific PCR, which targets the msp2 region, were found to be negative. Differences in prevalence were found to be statistically significant with regard to region, altitude, flock size, livestock production system, grazing system, presence of clinical cases and application of tick and tick‐borne diseases prophylactic measures. Kappa analysis revealed a poor concordance between microscopy and PCR (k = 0.14). Agreement with PCR is improved by considering microscopy and packed cell volume (PCV) in parallel. The prevalence of double infections was found to be 1.7, 2.5 and 24% for Anaplasma‐Babesia, Anaplasma‐Mycoplasma and Anaplasma‐Theileria, respectively. Co‐infection with three or more haemoparasites was found in 1.6% of animals examined. In conclusion, we demonstrate the high burden of anaplasmosis in small ruminants in Morocco and the high prevalence of co‐infections of tick‐borne diseases. There is an urgent need to improve the control of this neglected group of diseases.     

Factors associated with the prevalence of antibodies against Brucella abortus in water buffaloes from Santarém,Lower Amazon region,Brazil

We evaluated the factors associated with the prevalence of antibodies against Brucella abortus in buffaloes in the municipality of Santarém, Western Pará, northern Brazil. The study was conducted on 60 farms, representing 25.8% of the total buffalo farms in the region. From those farms, a total of 426 buffaloes were sampled, males of any age and females more than 24 months of age, to avoid a false‐positive reaction in the serological test due to vaccination. The Acidified Agglutination Serum Test was carried out on serum samples using B. abortus strain 1,119–3 as the antigen. Univariate and multivariate analysis were performed to investigate the association between brucellosis and potential risk factors. Of the 426 tested buffaloes, 29 were positive, resulting in an overall animal prevalence of antibodies against B. abortus at the animal level of 6.8% (4.6–9.6; 95% confidence interval). The herd level prevalence was 30% (18 of 60) and seroprevalence range within farms was from 0% to 100%. At the animal level, buffaloes raised in the floodplains tended (p = 0.06) to present a higher seroprevalence (9.70%) of antibodies against B. abortus than buffaloes raised in dry land (4.98%) and cows tended (p = 0.054) to have a higher seroprevalence than male buffaloes. Multivariate herd‐level analysis revealed association between farm type and brucellosis seroprevalence (p = 0.015); dairy farms were two times more likely to have seropositive buffalo than beef farms. Our survey demonstrated a high farm seroprevalence of B. abortus in buffalo raised in an Amazonian ecosystem with positive animals found in one third of sampled farms.     

Prevalence of Chlamydophila psittaci Infections in the Eyes of Cattle,Buffaloes, Sheep and Goats in Contact with a Human Population

This work is an example of cooperation between veterinary and human medicine being fully complementary and at the same time, indispensable to improve our knowledge on animal chlamydiosis. This study investigated the existence of ocular chlamydiae and determined the prevalence of its presence, chlamydiosis, in asymptomatic and diseased farm animals and adjacent humans. Data were obtained by the omp2 gene family Chlamydiaceae‐specific PCR. Two hundred cattle, buffaloes, sheep and goats and 44 human specimens were also examined. Conjunctival swabs from both the eyes were collected from all animals and humans using cotton swabs. Samples were tested for chlamydiae by Vero cells tissue culture, chicken embryo, modified Gimenez staining, direct fluorescein‐conjugated monoclonal antibody staining (FA), immunoperoxidase, CFT and PCR. The PCR‐RFLP revealed that Chlamydophila psittaci demonstrated in the conjunctival samples of cattle (68% asymptomatic and 88% diseased), of buffalo (68% asymptomatic and 72% diseased), of sheep (68% asymptomatic and 80% diseased), of goat (76% asymptomatic and 92% diseased) and of humans (77% asymptomatic and 82% diseased). The Cp. psittaci was the only chlamydiae demonstrated in all of the ocular conjunctival samples, which confirms the prevalence of Cp. psittaci in this population of animals and adjacent humans. Statistically, the animal species factor was calculated and was found to be of no significance. Yet, there appeared to be a significant difference in the percentage of animal that tested positive using the different methods. Detection of Cp. psittaci in most samples confirms the prevalence of Cp. psittaci in this population of animals and adjacent humans.     

Evaluation of Coxiella burnetii Status in Dairy Cattle Herds with Bulk‐tank Milk Positive by ELISA and PCR

Bulk‐tank milk (BTM) samples are frequently used to evaluate the health status of dairy livestock. A large‐scale investigation carried out in BTM samples from dairy cattle herds from a Q fever‐endemic region in Northern Spain revealed a high degree of exposure to Coxiella burnetii. This study was aimed at assessing the value of BTM samples analysis as an indicator of the C. burnetii status in dairy cattle herds. Three herds with BTM samples positive for C. burnetii by ELISA and PCR were selected, and blood, faeces and individual milk and BTM samples were analysed by serology and PCR. In spite of the high antibodies titres found in BTM samples, only one of the three farms presented an active infection by C. burnetii, as revealed by the presence of bacterial DNA in vaginal mucus and in environmental samples collected in the calving area, a seroprevalence around 40% in heifers and the seroconversion rate observed in cows. Results obtained indicated that the analysis of BTM samples is a good epidemiological tool at the population level that can be used to discriminate between seropositive and seronegative herds, but at the herd level, additional tests are necessary to evaluate whether Q fever is a potential problem in the farm. When Q fever is suspected in a cattle herd, sera from a small group of 1‐ to 3‐year‐old animals need to be analysed to investigate recent contact with C. burnetii.     

First Molecular Identification and Genetic Characterization of Theileria lestoquardi in Sheep of the Maghreb Region

Theileria lestoquardi is the most prominent Theileria species in small ruminants that causes malignant theileriosis of sheep in Africa and Asia. In the present survey, blood samples and ticks were collected in Kebili (southern Tunisia) from 166 Queue Fine de l'Ouest sheep. Giemsa‐stained blood smears, immunofluorescent antibody test (IFAT) and PCR were performed. The DNA was extracted from blood and analysed by PCR targeting 18S rRNA gene of Theileria spp. and then sequenced. A total number of 140 ticks were collected from a total number of 166 sheep during the four seasons. The ticks belonged to two genera and 4 species; the most frequent tick was Hyalomma excavatum 84.3% (118/140) and then Rhipicephalus spp. 15.7% (22/140). Only two animals had positive Giemsa‐stained blood smears, and they were also positive by IFAT. The amplicons had 99.3 and 99.6% homology with the BLAST published T. lestoquardi amplicons. To our knowledge, this is the first report of T. lestoquardi in small ruminants within the Maghreb region.     

The red fox (Vulpes vulpes) as a potential natural reservoir of human cryptosporidiosis by Cryptosporidium hominis in Northwest Spain

Giardia duodenalis and Cryptosporidium spp. are ubiquitous intestinal protozoa that parasitize domestic and wild animals, as well as human beings. Due to their zoonotic potential, the objective of the present study was to determine the presence of these pathogens in the fox population (Vulpes vulpes) located in Northwest Spain. A total of 197 faecal samples from legally hunted foxes were collected in the autonomous region of Galicia. The presence of G. duodenalis and Cryptosporidium spp. was investigated by PCR‐based methods amplifying the small subunit ribosomal RNA (ssu rRNA) gene of the parasites. Attempts to genotype obtained positive samples were subsequently conducted at the glutamate dehydrogenase (gdh) and β‐giardin (bg) genes of G. duodenalis, and the 60 kDa glycoprotein (gp60) gene of Cryptosporidium. Giardia duodenalis and Cryptosporidium spp. were identified in 19 (9.6%) and 12 (6.1%) of the investigated samples, respectively. However, five Cryptosporidium species were detected at the ssu rRNA locus: C. hominis (33.4%, 4/12), C. canis (25.0%, 3/12), C. parvum (16.7%, 2/12), C. ubiquitum (8.3%, 1/12) and C. suis (8.3%, 1/12). An additional Cryptosporidium‐positive sample was identified at the genus level only. Typing and subtyping of Giardia‐ and Cryptosporidium‐positive samples were unsuccessful. The detection of C. hominis in wild foxes indicates the probable overlapping of sylvatic and domestic cycles of this parasite in rural settings. Besides, this finding raises the question of whether red foxes may act as natural reservoirs of C. hominis. The detection of C. parvum and C. suis is suggestive of active transmission events between farm and wild animals, opening up the possibility of transmission to human beings.     

Brucella canis infection in dogs from commercial breeding kennels in Brazil

Canine brucellosis caused by Brucella canis is a neglected zoonosis worldwide and is a leading cause of reproductive failure in dogs, often causing substantial economic losses in breeding kennels. This study aimed to investigate the occurrence of B. canis infection in dogs of commercial breeding kennels located in São Paulo State, Brazil. A total of 753 dogs (183 males and 570 females) from 38 commercial kennels were clinically examined, and blood samples were collected for brucellosis diagnosis through blood culture. The association between clinical manifestations suggestive of brucellosis and positive results through blood culture was determined. Of the 753 dogs tested, 166 (22.0%) had at least one clinical sign suggestive of brucellosis and 158 (20.9%) had positive blood cultures. Seventy‐two dogs had positive blood culture and had at least one clinical sign suggestive of brucellosis, while 91 dogs showed at least one clinical manifestation suggestive of brucellosis although blood culture was negative. Of the 38 kennels, 16 (42.1%) had at least one positive dog. The prevalence of infection in each kennel varied from 3.8% to 62.6%. Abortion/stillbirth, failure to conceive and enlargement of lymph nodes were significantly associated with brucellosis in female. No association of clinical signs and positive results in blood culture was observed in males. None of the kennels has been carrying out programmes to control brucellosis, and the sale of infected dogs was considered a common practice yielding risks to the public health, in view of the zoonotic potential of the infection.     

Molecular Survey on Brucellosis in Rodents and Shrews – Natural Reservoirs of Novel Brucella Species in Germany?

Brucellosis is a widespread zoonotic disease introduced from animal reservoirs to humans. In Germany, bovine and ovine/caprine brucellosis were eradicated more than a decade ago and mandatory measures in livestock have been implemented to keep the officially brucellosis‐free status. In contrast, surveillance of wildlife is still challenging, and reliable data on the prevalence of brucellae in small mammal populations do not exist. To assess the epidemiology of Brucella spp. in rodents and shrews, a molecular survey was carried out. A total of 537 rodents and shrews were trapped in four federal states located throughout Germany and investigated for the presence of Brucella. Using a two‐step molecular assay based on the detection of the Brucella‐specific bcsp31 and IS711 sequences in tissue samples, 14.2% (n = 76) of the tested animals were positive. These originated mainly from western and south‐western Germany, where preliminary analyses indicate population density‐dependent Brucella prevalence in voles (Myodes glareolus) and mice (Apodemus spp.). recA typing revealed a close relationship to a potentially novel Brucella species recently isolated from red foxes (Vulpes vulpes) in Austria. The molecular detection of brucellae in various rodent taxa and for the first time in shrew species shows that these animals may be naturally infected or at least have a history of exposure to Brucella spp.     

Bovine anaplasmosis and tick‐borne pathogens in cattle of the Galapagos Islands

A cross‐sectional study was conducted to determine the species of Anaplasma spp. and estimate its prevalence in cattle of the three main cattle‐producing Galapagos Islands (Santa Cruz, San Cristóbal and Isabela) using indirect PCR assays, genetic sequencing and ELISA . Ticks were also collected from cattle and scanned for 47 tick‐borne pathogens in a 48 × 48 real‐time PCR chip. A mixed effects logistic regression was performed to identify potential risk factors explaining Anaplasma infection in cattle. A. phagocytophilum was not detected in any of the tested animals. Genetic sequencing allowed detection of A. platys ‐like strains in 11 (36.7%) of the 30 Anaplasma spp.‐positive samples analysed. A. marginale was widespread in the three islands with a global between‐herd prevalence of 100% [89; 100]_95%CI and a median within‐herd prevalence of 93%. A significant association was found between A. marginale infection and age with higher odds of being positive for adults (OR = 3.3 [1.2; 9.9]_{95% Bootstrap CI} ). All collected ticks were identified as Rhipicephalus microplus. A. marginale , Babesia bigemina , Borrelia theileri and Francisella ‐like endosymbiont were detected in tick pools. These results show that the Galapagos Islands are endemic for A. marginale .     

Investigation on the incidence of adverse reactions,viraemia and haematological changes following field immunization of cattle using a live attenuated vaccine against lumpy skin disease

The present study was performed to investigate the clinical impact and certain virological and haematological parameters following immunization of cattle against lumpy skin disease (LSD ). The study was conducted in a dairy cattle farm (215 animals), immunized with a Neethling strain‐based live vaccine. Twenty‐seven animals (14 lactating cows, four dry cows and nine calves) were randomly selected for repetitive blood and saliva samplings. An EvaGreen‐based real‐time PCR was designed to differentiate vaccine from field LSDV s. Vaccinated animals underwent examination for adverse reactions. Nodule samples were collected from two representative cases for histopathological testing and virus identification. Milk yield was calculated based on bulk‐tank measurements of all lactating cows (79). Viral DNA was detected between days 6–15 post‐vaccination (p.v.) at 63% of the sampled animals (17/27). Saliva and bulk‐tank milk samples were LSDV ‐negative. Pronounced swelling was observed at injection sites of 12% of the immunized animals (26/215), starting at day 6 p.v., and was resolved after 2–4 days. Small‐sized (<0.5 cm) cutaneous lumps were developed between days 8–18 p.v. at 9% of the vaccinated animals (19/215). These were observed in adult cows and not in calves/heifers. Resolution was observable 10 days post‐development. The vaccine virus was also identified in nodules and injection‐site aspirates. Haematological changes (e.g., lower leucocyte counts) were observed in cows and not in calves. Daily milk production was being reduced during the first 12 days p.v. LSD immunization of cows resulted in nodules and low viraemia levels. The fact that nodules and haematological changes were not observed in calves, along with the low viraemia, supports the reduced virulence of the Neethling vaccine strain. The characteristic nodules in vaccinated animals could allow clinical differentiation from those observed in LSD . The developed real‐time PCR efficiently differentiates infected from vaccinated cattle, and should be further validated as a tool in LSD surveillance.     

Molecular Demonstration of Trypanosoma evansi and Trypanosoma lewisi DNA in Wild Rodents from Cambodia,Lao PDR and Thailand

In this study, we investigated the molecular evidence of Trypanosoma evansi in wild rodents from Cambodia, Lao PDR and Thailand. Between November 2007 and June 2009, 1664 rodents were trapped at eight sites representative of various ecological habitats. Of those animals, 94 were tested by direct microscopic blood examination, 633 using the Card Agglutination Test for Trypanosomes (CATT/T. evansi) and 145 by Polymerase Chain Reaction (PCR) with two sets of primers: TRYP1 (amplifying ITS1 of ribosomal DNA of all trypanosomes) and TBR (amplifying satellite genomic DNA of Trypanozoon parasites). Using TRYP1, based on the size of the PCR products, 15 samples from the three countries were positive for Trypanosoma lewisi (two were confirmed by sequencing), and three were positive for Trypanozoon (one was confirmed by sequencing and three by TBR primers); the specificity of the primers failed as rodent DNA was amplified in some cases. Using TBR, six samples were positive for Trypanozoon (one was confirmed by sequencing); as T. evansi is the only species of the Trypanozoon sub‐genus possibly present in Asian rodents, these results confirmed its presence in rodents from Thailand (Rattus tanezumi) and Cambodia (R. tanezumi, Niviventer fulvescens & Maxomys surifer). Further investigations are necessary to establish the situation in Lao PDR. None of the 16 samples most strongly positive to the CATT proved to be positive for Trypanozoon by PCR. The merits of the CATT for such studies were not confirmed. Studying the urban and rural circulation of these parasites in rodents will enable an evaluation of human exposure and infection risk, as human infections by T. evansi were recently described in India and by T. lewisi in India and Thailand. As sequencing PCR products is expensive, the development of new molecular and serological tools for rodents would be very useful.     

Persistence of Mycobacterium bovis bacillus Calmette–Guérin (BCG) Danish In White‐tailed Deer (Odocoileus virginianus) Vaccinated with a Lipid‐Formulated Oral Vaccine

Mycobacterium bovis, the causative agent of tuberculosis in animals, has a broad host range, including humans. Historically, public health concerns prompted programs to eradicate tuberculosis from cattle in many nations. Eradication efforts decreased the prevalence of bovine tuberculosis; nevertheless, some countries encountered significant obstacles, not least of which was a wildlife reservoir of M. bovis. Efforts to decrease the size of the affected wildlife populations have neither eliminated disease nor eliminated transmission to cattle. Consequently, the use of a vaccine for wildlife is being explored. The vaccine most studied is M. bovis BCG, an attenuated live vaccine, first developed 100 years ago. The most efficient and effective means of vaccinating wildlife will be an oral vaccine. White‐tailed deer in Michigan, USA, constitute a reservoir of M. bovis. White‐tailed deer are a popular game species, and as such, represent a food animal to many hunters. BCG persistence in deer tissues could result in human exposure to BCG. Although non‐pathogenic, BCG exposure could induce false‐positive skin test results, confounding the central component of public health surveillance for TB. The objective of the present study in white‐tailed deer was to evaluate persistence of lipid‐encapsulated BCG and a liquid suspension of BCG after oral administration at two different dosages. Vaccine was not recovered at any time after oral consumption of a bait containing a single dose (1 × 10⁸ CFU) of lipid‐encapsulated BCG. However, persistence was consistent in deer consuming 10 lipid‐encapsulated baits (1 × 10⁹ CFU), with BCG recovered from at least one deer at 1, 3, 6, 9 and 12 months after consumption. Persistence of up to 9 months was seen in deer vaccinated with orally with a liquid suspension. Persistence of BCG was limited to lymphoid tissue and never found in samples of muscle collected at each time point. Although the risk of exposure to hunters is low, BCG persistence should be considered prior to field use in white‐tailed deer.     

A multifaceted risk model of brucellosis at the human–animal interface in Egypt

Brucellosis is a highly contagious zoonosis affecting humans and a wide range of domesticated and wild animal species. An important element for effective disease containment is to improve knowledge, attitudes and practices (KAP) of afflicted communities. This study aimed to assess the KAP related to brucellosis at the human–animal interface in an endemic area of Egypt and to identify the risk factors for human infection. A matched case–control study was conducted at the central fever hospitals located in six governorates in northern Egypt. Face‐to‐face interviews with cases and controls were conducted using a structured questionnaire. In total, 40.7% of the participants owned farm animals in their households. The overall mean practice score regarding animal husbandry, processing and consumption of milk and dairy products were significantly lower among cases compared with controls (−12.7 ± 18.1 vs. 0.68 ± 14.2, respectively; p < .001). Perceived barriers for notification of animal infection/abortion were predominate among cases and positively correlated with participants’ education. The predictors of having brucellosis infection were consumption of unpasteurized milk or raw dairy products and practicing animal husbandry. Applying protective measures against infection significantly reduced its risk. A model predicting risk factors for brucellosis among those who own animal showed that frequent abortions per animal increased the chance for brucellosis infection among human cases by 50‐fold (95% CI: 8.8–276.9), whereas the use of protective measures in animal care reduced the odds (OR = 0.11 [95% CI: 0.03–0.45]). In conclusion, consumption of unprocessed dairy products was equally important as contact with infected/aborted animals as major risk factors for Brucella spp. infection among humans in Egypt. There is poor knowledge, negative attitudes and risky behaviours among villagers which can perpetuate the risk of brucellosis transmission at the human–animal interface. This supports the need for integrating health education into the national brucellosis control programme.     

Molecular Survey of Anaplasma and Ehrlichia of Red Deer and Sika Deer in Gansu,China in 2013

Anaplasma and Ehrlichia are important emerging tick‐borne pathogens in both humans and animals. Here, we conducted a molecular surveillance study in Gansu, China to assess the prevalence of Anaplasma and Ehrlichia spp. in red deer and sika deer based on polymerase chain reaction (PCR) analysis and sequencing of 16S rRNA or msp genes. PCR revealed that the prevalence of Anaplasma ovis, Anaplasma bovis and Anaplasma platys of the Qilian Mountain samples was 32%, 9% and 9%, respectively; the prevalence of Anaplasma ovis, Anaplasma bovis, Anaplasma platys was 20%, 15% and 15% among the Long Mountain samples, respectively. Of the Long Mountain samples, two (5%) of the 40 samples were positive for Ehrlichia canis, but all 44 of the Qilian Mountain samples were negative for E. canis, and no other Anaplasma or Ehrlichia spp. were found in the samples. The phylogenetic tree showed that the newly isolated Anaplasma and Ehrlichia spp. could be classified as belonging to four clades, including an A. bovis cluster, A. ovis cluster, A. platys cluster and E. canis cluster. In addition, Bartonella schoenbuchensis was firstly identified in blood samples from red deer in Gansu, China. Our results provide important data to increase the understanding of the epidemiology of anaplasmosis and ehrlichiosis of red deer and sika deer and will assist with the implementation of measures to control anaplasmosis and ehrlichiosis transmission to red deer, sika deer and other animals in Gansu, China.     

Exploratory investigation of Q fever in apparently healthy meat sheep flocks in Belgium

Q fever is a cosmopolitan disease affecting both humans and many animal species. Although sheep are often implicated in human Q fever outbreaks, the disease remains largely underestimated in meat sheep flocks. In order to fulfil this gap, a preliminary study was performed aiming to investigate the serological and molecular aspects of infection with Coxiella burnetii among meat sheep flocks in Belgium. Five Belgian sheep flocks were recruited for this work. Indirect ELISA was used, and in addition, real‐time PCR was performed on samples of milk, rectal and vaginal swabs, to understand the dynamics of bacterial shedding. Despite the low overall apparent seroprevalence of 1.39% (95% CI : 0.04–7.5), a high rate of bacterial shedding was found, with 27.7% of tested sheep (N  = 72) with a positive result to PCR , especially through the rectal and vaginal routes and in seronegative animals. Furthermore, Coxiella burnetii DNA was detected in 26.76% of seronegative animals. It can be concluded that an overall good clinical condition of the sheep cannot be used to exclude the presence of C. burnetii in a flock. Furthermore in the diagnosis of Q fever in sheep, serology alone was not a sensitive diagnostic tool. On the contrary, molecular biology allowed to detect bacterial shedding, which is an essential element in order to assess the risk due to the contact with shedding animals. At the light of these results, the role of meat sheep flocks in the epidemiology of Q fever in Belgium needs to be better understood.     

Bovine Brucellosis in Argentina and Bordering Countries: Update

Bovine brucellosis is a zoonotic disease spread worldwide. The infection in cattle is predominantly caused by Brucella abortus and is usually detected in pregnant females through abortions. The disease is endemic in Argentina; however, infection in humans is underestimated and often not reported. The prevalence of bovine brucellosis in countries bordering Argentina is quite variable: 0.04% in Uruguay, 10.20% in the north and 0.06% in the south of Brazil, 0.2% in Chile, 3.15% in Paraguay and 2.27% in Bolivia. In 1999, the Argentine National Control and Eradication Program was implemented. Its strategies include identification of vaccinated animals, compulsory vaccination with B. abortus S19 of 100% of 3‐ to 8‐month‐old females, negative serological tests before animal movements and categorization of farms in terms of their brucellosis status. The epidemiological surveillance in milk is performed through the milk ring test and the indirect ELISA. The result of a national brucellosis survey performed in 2004 indicates that 12.4% (95% CI: 10.89–14.0) of Argentine beef farms are seropositive to Brucella and that the apparent prevalence in cattle is 2.10% (95% CI: 1.90–2.40). The official serological diagnostic tests are as follows: buffered plate antigen test, as screening, serum agglutination test, 2‐mercaptoethanol and fluorescence polarization assay, competitive ELISA, as confirmatory tests, and complement fixation test, as definitive test. Santa Fe and a district in Córdoba have ‘Outstanding Plans’. Tierra del Fuego is a ‘Zone free from bovine brucellosis’. One question arising when studying the Argentine situation is why the disease remains endemic if good regulations exist to control and eradicate it. In future, some different aspects might be evaluated to understand it, and further studies should be performed to prioritize, select and refine control strategies.