An abortion case in a dromedary camel caused by a B. melitensis strain belonging to the East-Mediterranean genetic group (GT42 by multiple-locus variable-number tandem-repeat analysis (MLVA8) and GT108 by MLVA11) is reported. Earlier, the dromedary was seronegative for Brucellosis for 6 years. The animal got infected between 112 and 204 days of gestation from an unknown source, developed bacteriaemia and the acute infection resulted in the abortion at 249 days of gestation. The pathological findings in the foetus and the placenta were not pathognomonic for Brucellosis. In the foetus, both pleural and abdominal cavities were filled with bloody fluid and pleurisy was observed. In the placenta, oedema, diffuse mineralisation and focal detachment of the trophoblast were detected without inflammatory changes. The Brucella pathogens were excreted to the environment with the aborted foetus and the placenta. The bacteria disappeared from the blood and the uterus of the dam within a short time after abortion and were isolated only from some of the lymph nodes. The Brucella seropositive camel in its acute phase of the disease did not infect other contact animals. In addition, the transmission of the disease to other dromedaries at the time of abortion was also prevented with appropriate biosecurity measures.

A vaccination experiment was conducted in dromedary camels at CVRL. Six adult dromedaries received 2 ml of the Brucella abortus vaccine RB51 subcutaneously and 6 camels 1ml of the B. melitensis Rev 1. Four different serological tests were conducted to follow the seroconversion post vaccination. None of the 6 dromedaries vaccinated with RB51 produced antibodies, measured for 1 year, due to lack of lipopolysaccharide O - side chains in the vaccine. However, antibodies appeared in all 6 dromedaries 30 days post vaccination with B. melitensis Rev 1, which declined slowly over time. Three hundred thirty days post vaccination ELISA antibodies were still present in 5 of the 6 dromedaries as well as RBT, SAT and CFT antibodies in some of them. This is a disadvantage as all serological methods for the diagnosis of brucellosis cannot differentiate between vaccine or natural infection titres.

Fifteen non pregnant female dromedaries of different age were intra-tracheally and intra-nasally infected with a Brucella melitensis strain belonging to the genetic group East Mediterranean (former African group). The development of Brucella antibodies in the infected dromedaries was investigated over a period of 12 months by comparing 15 different serological tests. Three uninfected control dromedaries remained negative during the entire experiment. Our investigations revealed that only 2 serological tests were characterised by a high degree of sensitivity for the diagnosis of brucellosis in dromedaries. These two tests are a Rose Bengal Test (RBT) from Vircell, Spain and a competitive ELISA (cELISA) from Ingenasa, also from Spain. The blood culture revealed that all tested sodium citrate blood samples were negative. Results of the enriched citrate blood samples of the infected 15 dromedaries fluctuated and became positive for a short time, but at the end of the experiment only one remained positive. Direct examination by PCR did not show any evidence of the pathogen in the EDTA blood of the 15 infected dromedaries. Also, all nasal swabs taken up to a period of 3 months post infection remained negative in both PCR and microbial culture.

The present study describes the laboratory investigations after a single right eye drop (3.1 x 10(9) CFU live bacteria) immunisation of 6 dromedary camels (Camelus dromedarius) with live attenuated B. melitensis Rev 1 vaccine. The experiment was conducted over a period of 5 months. The vaccine strain was isolated for 16 days from only the right eye of the vaccinated dromedaries, but not from the left eye and both nostrils. Similar pattern of results was obtained by polymerase chain reaction. It was negative for the left eye, both nostrils (except for one dromedary) and for EDTA blood and serum. All vaccinated dromedaries seroconverted from day 16 after vaccination until 4 months shown by Rose-Bengal test and slide-agglutination test. No serological reactions were found after 5 months. The complement fixation test remained negative throughout the experiment. Information about the vaccination against brucellosis in camels, the within host disperse of the vaccine strain and the serological response are scarce. The experiment provided basic data about the feasibility of Brucevac conjunctival vaccine in camels. However, to prove if the immunised dromedaries acquired a lifelong immunity against brucellosis, pregnant vaccinated dromedaries need to be challenged with a field B. melitensis strain. We also recommend changing the conjunctival vaccination route to subcutaneous or intramuscular to prevent accidental infection due to B. melitensis vaccine strain excreted by lacrimation.

Four female and 1 male dromedaries serologically positive for Brucellosis, were treated 3 times with enrofloxacin and Brucella IgGs, intravenously over a time period of 3 months. Despite this treatment, the brucellosis positive dromedaries remained serologically positive.

Three 8-12 year old milking dromedaries were tested serologically positive several times for brucellosis with the RBT, SAT and CFT over a period of 4 months. One animal excreted the brucella organisms in low numbers through the milk. It was decided to euthanise all 3 animals. All 3 camels had delivered healthy male calves 3 months earlier and the dams themselves were in very good bodily condition. After euthanasia, a thorough bacteriological and histological investigation of all organs, lymph nodes, joints and milk was carried out. Special attention was directed towards the mammary glands and adjacent lymph nodes, as well as, towards the reproductive tract. For the isolation of brucella microorganisms, a routine and a concentration method were applied.Brucella melitensis, serovar 3 was isolated from all 3 dromedaries. Histopathological investigations showed no lesions in any of the organs except in lymph nodes, from which B. melitensis was isolated. The lymph nodes showed marked sinusoidal oedema, activated follicles and histiocytosis. None of the 3 camels harboured brucella bacteria in their reproductive tracts or adjacent lymph nodes. From only one dromedary, which excreted B. melitensis through the milk, were brucella organisms isolated, from the right and left udder cisterns. B. melitensis bacteria were isolated from Lnn cervicales superficiales in all 3 cases and from other different lymph nodes, but not in all 3 cases. The concentration method is essential for the recovering of brucella organisms from organs and milk.

A role for bacteriophage therapy was envisaged earlylast century; however, due to discovery of the antimicrobials, it fell out of research interest. Currently, bacteriophages are resurfacing as an alternative to antimicrobials in order to overcome the increasing incidence of antimicrobial resistance. Here, we report isolation of bacteriophages against Escherichia coli, Shigella spp., Aeromonas hydrophila and Citrobacter sedlakii isolates of equine origin. Phages were isolated from equine farm soil and sewage samples. For enrichment, sample aliquot was incubated overnight with host bacteria at 37 C with vigorous shaking. The crude lysate obtained was centrifuged and filtered and the presence of any phage in the suspension was detected by agar overlay technique. Appropriate dilutions of enriched samples were plated to obtain individual plaques and the most dominant plaque was transferred into SM buffer, serially diluted and plated for plaque re-isolation three times to ensure purity, followed by large scale preparation of phage stocks. Any host nucleic acids was degraded using pancreatic DNaseI and RNase and bacteriophage particles were precipitated using PEG8000. Phage titre was determined by plaque assay and phage concentrates were accessioned in the Veterinary Type Culture Collection (VTCC) repository. The phage concentrates were visualized by transmission electron microscopy (TEM). The temperature stability of bacteriophages was checked after incubating phage concentrates over the range of 4 C-8 0C temperature for one hour. A clear single plaque was obtained on nutrient agar against Shigella spp. and after purification and concentration, its analysis by electron microscopy revealed presence of multiple phages belonging to families Myoviridae, Siphoviridae and Podoviridae. However in case of Citrobacter sedlakii, a Siphoviridae phage (VTCCBPA61) with dimensions: 60 nm x 650 nm was observed. Against a pathogenic A. hydrophila isolate of equine origin (expressing aerolysin gene), a Myoviridae phage (VTCCBPA6) was isolated with dimensions: 62 nm x 138 nm. Against E. coli of equine origin, a Myoviridae phage (VTCCBPA9) of dimensions: 86 nm x 100 nm was obtained. Bacteriophage VTCCPBA61 against C. sedlakii was found to completely lose its biological activity at 65 C in vitro however the group of phages against Shigella spp. were found to be stable upto temperature as high as 80C. Thus we demonstrated the basic biological characteristics of phages, and some novel ones (such as against A. hyrophila and C. sedlakii) bacterial isolates of equine originwhich have never been reported till now. These lytic phages could find potential in phage therapy, as biocides, in biosensors and in phage ligand technology and are being explored by us further to depict their therapeutic value in small animal model. As more studies are reporting safety, tolerance and efficacy of phage therapy in humans and animals, their use in phage therapy has a promising future as an emerging alternative to chemical agents.

The results of two laboratory tests, PCR and culture were compared with each other for the recovery of Brucella melitensis and Brucella abortus from spiked camel, goat and sheep milk samples. The results showed the same sensitivity for camel milk but a lower PCR sensitivity for sheep and goat milk.

The teat canals and mammary gland cisterns of 30 necropsied omniparous non-lactating dromedaries aged between 8 and 20 years were microbiologically investigated. In total 242 teat canals and 242 cisterns were swabbed from which 36 different bacterial species comprising 17 families were isolated. In total 24% teat cabals and 22% mammary cisterns harboured bacteria colonies but they grew only in very low numbers of between 1 to 13 colonies per agar plate. It can be stated that our investigation revealed that a high number of dromedary glands are either sterile or harbour only a low number of mainly non-mastitis pathogens.

Although camelids are an important domestic species with more than 25 million members, little is known about vaccine adjuvant efficacy, safety and mechanism of action in this species. This presents a major problem for design of effective camelid vaccines. This is of more than theoretical interest given the recent emergence of camels as vectors of transmission to humans of lethal viral diseases such as Middle Eastern Respiratory Syndrome (MERS) coronavirus. Hence availability of well-validated camelid vaccine adjuvants may be important not just for vaccines to prevent diseases of camels but also to block their ability to transmit disease to humans. In this study, we used dromedaries to test the safety and efficacy of four different adjuvant formulations (Advax™ HCXL, Advax AF-1, Advax AF-2 or alum) together with four different antigen formulations (B. mallei, C. pseudotuberculosis, C. perfringens, Rhinovirus) administered by subcutaneous injection in the neck region of adult animals. All the Advax delta inulin-based adjuvants and the alum adjuvant were well tolerated, with no severe lesions such as the draining granulomas that are caused by oil emulsion adjuvants. There was no trend for increased vaccine reactogenicity in camels that had existing immunity to the vaccine antigens. Overall, the vaccines had modest immunogenicity in these adult animals indicating the need for further research to identify the optimal adjuvant formulation, dose and immunisation route for camelid vaccines. A case of dystocia in dromedary camel due to posterior presentation of foetus with flexion of hind leg was reported. Dead foetus was delivered per-vagina by traction.

In 2013, a novel betacoronavirus was identified in fecal samples from dromedaries in Dubai, United Arab Emirates. Antibodies against the recombinant nucleocapsid protein of the virus, which we named dromedary camel coronavirus (DcCoV) UAE-HKU23, were detected in 52% of 59 dromedary serum samples tested. In an analysis of 3 complete DcCoV UAE-HKU23 genomes, we identified the virus as a betacoronavirus in lineage A1. The DcCoV UAE-HKU23 genome has G+C contents; a general preference for G/C in the third position of codons; a cleavage site for spike protein; and a membrane protein of similar length to that of other betacoronavirus A1 members, to which DcCoV UAE-HKU23 is phylogenetically closely related. Along with this coronavirus, viruses of at least 8 other families have been found to infect camels. Because camels have a close association with humans, continuous surveillance should be conducted to understand the potential for virus emergence in camels and for virus transmission to humans.

Background Glanders, caused by the gram-negative bacterium Burkholderia mallei, is a highly infectious zoonotic disease of solipeds causing severe disease in animals and men. Although eradicated from many Western countries, it recently emerged in Asia, the Middle-East, Africa, and South America. Due to its rareness, little is known about outbreak dynamics of the disease and its epidemiology. Methodology/Principal Findings We investigated a recent outbreak of glanders in Bahrain by applying high resolution genotyping (multiple locus variable number of tandem repeats, MLVA) and comparative whole genome sequencing to B. mallei isolated from infected horses and a camel. These results were compared to samples obtained from an outbreak in the United Arab Emirates in 2004, and further placed into a broader phylogeographic context based on previously published B. mallei data. The samples from the outbreak in Bahrain separated into two distinct clusters, suggesting a complex epidemiological background and evidence for the involvement of multiple B. mallei strains. Additionally, the samples from Bahrain were more closely related to B. mallei isolated from horses in the United Arab Emirates in 2004 than other B. mallei which is suggestive of repeated importation to the region from similar geographic sources. Conclusion/Significance High-resolution genotyping and comparative whole genome analysis revealed the same phylogenetic patterns among our samples. The close relationship of the Dubai/UAE B. mallei populations to each other may be indicative of a similar geographic origin that has yet to be identified for the infecting strains. The recent emergence of glanders in combination with worldwide horse trading might pose a new risk for human infections.

The described trial substantiated the observation of physicians and parents, that after consuming pasteurised camel milk on a regular basis, a positive effect on impairments of various nature and proportions appeared to be apparent amongst a group of children with Autism Spectrum Disorder (ASD) symptoms or related neurological pathogenesis. Fourteen days after the consumption of 500 ml of pasteurised camel milk, the probands exhibited regular bowel movements and five of eight probands developed a normal sleep pattern. The overall observation revealed also a decreased hyperactivity, increased alertness, better social interaction and many parents observed a newly expressed effort of their children to obey instructions. However, there was no difference in the level of b-casomorphin-7 excretion in the urine of probands and controls using a non-commercial ELISA kit. The reason for this unexpected result is explained.

A natural infection of a glanders case in a dromedary camel (Camelus dromedarius) is reported here for the first time. This animal was most probably infected by glanderous horses which were housed nearby.The camel showed the classical clinical signs of glanders with fever, emaciation, mucopurulent discharge from the nose, and glanderous lesions in the lung, choanae and nasal septae. The serum was positive in the CFT and cELISA.B. mallei was isolated from the EDTA blood, from the choanal lesions as well as from nasal pus swabs after enrichment in guinea pig (Strauss reaction). All samples were positive for B. mallei with the PCR but negative for B. pseudomallei.

We confirm a natural infection of dromedaries with glanders. Multi locus variable number tandem repeat analysis of a Burkholderia mallei strain isolated from a diseased dromedary in Bahrain revealed close genetic proximity to strain Dubai 7, which caused an outbreak of glanders in horses in the United Arab Emirates in 2004.