Pathogenic Escherichia coli causes infections responsible for economic losses in animal herds worldwide. Although this bacterium is well studied in livestock and poultry, studies of camelid infections caused by extraintestinal pathogenic E. coli (ExPEC) are limited. In this study, a set of ExPEC from camel carcasses (n = 150) was characterized with respect to phylogenetic groups, 162 O serotypes, and 35 virulence-associated genes (VAGs) using PCR screening. ExPEC frequently belonged to phylogroup B1 (58.7%), followed by phylogroups C, A, and B2 (12.7%, 12.0%, and 9.3%, respectively). Additionally, the set of ExPEC contained 36 different serotypes. The ExPEC isolates were found to typically encode ≥5 tested VAGs, particularly those related to adhesion (afaI, fimA, pap, sfa, tsh), iron acquisition (fyuA, iroN, iucC, sitA), host cell damage (α-hly, cdt, cnf1, sat), invasion (ibeA), and bacterial protection (iss, ompT, traT). Moreover, ExPEC from camel adults and calves were different from each other. Among isolates from calves, prevalence was significantly higher for phylogroups C (q < 0.001) and E (q < 0.01), and ten VAGs, including fitness factors (eitA, fepC, fyuA, iroN, iss, iucD, ompT, sitA), as well as VAGs with stronger link to pathogenicity (hlyF, and pap). To identify potential reservoir of camel ExPEC strains, fecal E. coli (n = 139) from healthy camels were also analyzed. Based on the identified characteristics, ExPEC were distinguishable from fecal isolates of healthy camels, suggesting an exogenous source of ExPEC infections, likely transmitted from wild birds and human keepers.

African Horse Sickness (AHS) is a devastating vector-borne viral disease of equids with a mortality up to 95 % in naïve domestic horses. The causative African horse sickness virus (AHSV) is a distinct species of the genus Orbivirus of the family Sedoreoviridae, consisting of nine serotypes showing limited cross protection. AHSV is transmitted by Culicoides biting midges. Outbreaks cause huge economic losses in developing African countries. AHS has become a serious threat for countries outside Africa, since endemic Culicoides species in moderate climates appear competent vectors of the closely related prototype orbivirus, bluetongue virus. In the developed world, AHS outbreaks will result in losses in the equestrian industry and will have an enormous emotional impact on owners of leisure horses. Live-attenuated vaccine viruses (LAVs) are unsafe and differential detection of infected equids in LAV-vaccinated populations is not possible. Reverse genetics has paved the way to develop improved AHS vaccines, particularly with regard to vaccine safety and the DIVA principle (DIVA = Differentiating Infected from Vaccinated). Here, we developed an AHS Disabled Infectious Single Animal (DISA)-DIVA vaccine platform based on a dispensable deletion in genome segment 10 completely abolishing its virulence. The vaccine platform was applied for all nine AHSV serotypes by exchange of genome segments 2 and 6 encoding serotype specific immunodominant outer shell proteins. These nine promising AHS DISA-DIVA vaccines, named shortly DISA1 to DISA9 after their serotype, were extensively checked by several in vitro methods. The accompanying DIVA PCR-test targeting the deleted region in genome segment 10 was developed and validated. Based on previous research on virulent AHSV, DISA1 to DISA9 are not virulent, are not transmittable by midges, can be distinguished from wildtype AHSV, and are now ready for vaccination-challenge experiments in the equine target host to study efficacy.

Epizootic lymphangitis (EL) is a highly prevalent and contagious infectious disease affecting horses in many parts of Ethiopia caused by Histoplasma capsulatum sensu lato ('var. farciminosum'). In this study, 12 suspected isolates of H. capsulatum sensu lato or yeasts unidentified by conventional biochemical tests isolated from Ethiopian horses with EL were characterised by internal transcribed spacer sequencing. Six of the 12 isolates were identified to be members of H. capsulatum sensu lato and the other six were Pichia kudriavzevii (synonym: Candida krusei) (n = 3), Trichosporon asahii (n = 1), Geotrichum silvicola (n = 1) and Moesziomyces aphidis (n = 1), respectively. The six H. capsulatum sensu lato isolates were further characterised by multilocus sequence analysis. Four distinct gene loci (arf [462 bases], H-anti [410 bases], ole1 [338 bases] and tub1 [272 bases]) of these six isolates as well as those of two H. capsulatum sensu lato ('var. farciminosum') reference strains (ATCC 58332 and ATCC 28798) were polymerase chain reaction (PCR)-amplified and sequenced. Phylogenetic analyses of their concatenated nucleotide sequences showed that three of the isolates and the reference strain ATCC 58332 were identical and belonged to the Eurasia clade within Latin American (LAm) A (H. suramericanum), and those of the other three isolates and the reference strain ATCC 28798 were identical and belonged to the Africa clade. At least two distinct phylogenetic clades of H. capsulatum sensu lato were circulating in Ethiopian horses with EL. Advanced molecular technologies and bioinformatics tools are crucial for the accurate identification and typing of pathogens as well as the discovery of novel microorganisms in veterinary microbiology. Using multilocus sequence analysis with four concatenated housekeeping gene loci, at least two distinct phylogenetic clades, namely Eurasia clade and Africa clade, of Histoplasma capsulatum sensu lato were confirmed to be circulating in Ethiopian horses with epizootic lymphangitis.

Chlamydiaceae are a family of obligate intracellular bacterial pathogens that affect both humans and animals. Recently, a new species named Chlamydia (C.) buteonis was isolated from hawks. In this study, we aimed to investigate the prevalence of Chlamydiaceae in 60 falcons that underwent a routine health check at a specialized clinic in Dubai, United Arab Emirates. Using real-time PCR, we analyzed cloacal and tracheal swabs from these birds and found that 39 of them tested positive for Chlamydiaceae. Subsequent real-time PCR assays specific for C. psittaci, C. abortus, C. avium, and C. gallinacea yielded negative results, while testing positive for C. buteonis. Analysis of ompA and MLST sequences indicated a highly conserved group of strains within this set of samples, but with sequences distinct from the C. buteonis RSHA reference strains and other C. buteonis strains isolated from hawks in the United States. Two strains were further isolated by cell culture and sequenced using whole-genome sequencing, confirming the clustering of these falcon strains within the C. buteonis species, but in a separate clade from the previously identified hawk strains. We also developed a SNP-based PCR-HRM assay to distinguish between these different genotypes. Overall, our findings suggest a high prevalence of C. buteonis in falcons in Dubai and highlight the importance of monitoring this pathogen in birds of prey.

The aim of the present study was to examine the mammary gland of dromedary camels using ultrasonography, endoscopy and radiography. These techniques are easy to perform in the field and feasible to diagnose pathological conditions of the mammary gland. Udders of 49 slaughtered and 26 adult dromedary camels submitted for necropsy were used for the examinations. Additionally, 11 lactating female dromedary camels were selected for the ultrasonographic udder examination. The transition from the milk ducts into the udder cistern, the teat cistern and the teat canals were examined in individual udders. Teat cistern length, teat end width, teat wall thickness, teat cistern width and middle cistern wall thickness were measured using ultrasonography. The measurements resulted in mean values of the teat cistern length of 37.3 mm, the teat end width of 2.0 mm, the teat wall thickness of 4.4 mm, the teat cistern width of 8.2 mm and the cistern wall thickness of 3.5 mm. The teat wall was differentiated into three layers, a hyperechoic outer layer, a hypoechoic middle layer and a hyperechoic inner layer. The mid cistern wall was hyperechoic. Endoscopic examination is an easy to perform and practicable method for examining the inner structures of the teats of dead animals; however, the feasibility has not been shown in lactating animals yet. Ring-like folds were present in the teat cistern, which protruded horizontally into the lumen. It was also possible to visualize the branchlike transition of the teat cistern into the larger milk ducts. Radiographic examination using barium sulfate contrast medium showed that the teat cistern ends in a network of initially wide but branching and narrowing milk ducts. The two teat canals and cisterns are completely independent of each other and there is no communication between the glandular tissue of the two canals and cisterns.

The objectives of the present study were to describe specific anatomical structures of the dromedary udder. Eighty-six dromedary udders were examined, taking morphological measurements and producing injection casts using resin, gelatin, and paraffin. The udder of the dromedaries consists of four quarters. The shape of the udders and teats varies considerably between animals and is influenced by age, breeding, and lactation status. The most frequently found udder form was the globular udder (48.8%) and the most common teat form in this study was the funnel teat (44.2%). The most common teat tip shape was a smooth or a slightly rough ring teat (61.6%). Injection casts showed a complete separation of the teat canals. There is also no communication between tributary mammary complexes. Resin injections of the glandular tissue adjacent to the teat cistern showed an extensive branching into large, medium, and small milk ducts. Frozen sections of the udder revealed complete separation of the right and left mammary complex through the Sulcus intermammarius. The teat sections showed longitudinal folds from the tip of the teat to the base of the teat. A ring fold at the transition from the Ductus papillaris to the teat cistern was present. The results of this study increase the knowledge of the anatomical structures of the dromedary udder, which may be useful for breeding a selection of dairy dromedaries.

This article describes the development of the pathogenic dimorphic fungus Histoplasma capsulatum var. farciminosum (HCF), which is the causative agent of Equine epizootic lymphangitis (EEL), from the mycelial form in the soil to the yeast form in the horse. In this study, the stages and morphology of HCF were identified through histopathological analysis and culture with various samples collected in Ethiopia from 15 horses showing clinical signs of EEL. In equids, especially cart horses in Ethiopia, poor-quality harnesses cause cutaneous wounds, which often attract flies facilitating the transmission of the fungus. Also, HCF infection occurs through open wounds or ocular mucous membranes when horses roll on contaminated damp soil. Respiratory histoplasmosis can occur through inhaling fungal spores, which is rare. HCF microconidia enter the lungs and skin wounds and are phagocytized by tissue-resident macrophages. The spores undergo intracellular replication within the macrophages transitioning into yeasts. The infected macrophages undergo lysis releasing pathogenic yeast cells into the surrounding tissue. Consequently, yeast-rich purulent exudate is produced, contaminating the soil in stables where yeast cells germinate into the mycelial form, and the entire process starts from the beginning.

OBJECTIVES:To determine normal ocular parameters of the MacQueen's bustard (Chlamydotis macqueenii) and describe ophthalmic lesions in a captive bred population. ANIMALS STUDIED:Captive breeding population of 257 Macqueen's bustards. METHODS:All birds were screened for ocular abnormalities using direct ophthalmoscopy. Abnormalities were photographed. Normative values for Schirmer tear test-1 (STT-1), applanation tonometry, aerobic and anaerobic bacterial culture, fungal culture, and transcorneal ocular ultrasonography were derived from multiple cohorts of clinically normal adult birds. Five birds with ocular pathology also underwent transcorneal ultrasonography. Statistical comparisons for normative values between OD and OS, and males and females were made using a paired t-test or Mann-Whitney U-test, with a significance level of p < .05. RESULTS:Mean tear production based on Schirmer tear test 1 (STT-1) was 10.16 ± 4.61 mm/min (3-21 mm/min). Mean intraocular pressure (IOP) was 12.42 ± 4.94 mm Hg (5-26 mm Hg). Staphylococcus species were the most isolated bacteria from the conjunctival surfaces of normal birds (85%). Significant differences were found in transcorneal ultrasonographic measurements between males and females for axial globe length (p = .032), vitreous body depth (p = .049) and lens thickness (p = .0428). Corneal fibrosis was the most observed ocular abnormality amongst eyes with pathological changes (39%). CONCLUSIONS:Schirmer tear testing, tonometry and transcorneal ultrasound can easily be utilized in MacQueen's bustards and provide reproducible results. Normal parameters for these tests were determined, and common pathological ocular changes were described in this species.

At a visit to an unfenced desert conservation reserve in Dubai, United Arab Emirates in February 2022, severe skin disease was noted among resident Arabian oryx (Oryx leucoryx), manifesting as dark grayish to black bark-like thickened skin. Between and 45% and 60% of the oryx showed unrest and pruritus. Sarcoptes scabiei was detected at necropsy of six adult animals. Treatment with ivermectin-medicated pellets at 0.3 mg/kg estimated body weight over two periods of 7 d with a 14-d interval between treatments resulted in improved body and skin condition and hair regrowth. Although severe hyperkeratosis was still present shortly after treatment, no live Sarcoptes mites were found in parasitological examination of skin scrapings of two necropsied animals. By 4 mo post treatment the oryx had returned to normal body condition and coat condition.

Equine infectious anemia (EIA) is an important viral disease characterized by persistent infection in equids worldwide. Most EIA cases are life-long virus carriers with low antibody reactions and without the appearance of clinical symptoms. A serological test with high sensitivity and specificity is required to detect inapparent infection. In this study, a B-cell common epitope-based blocking ELISA (bELISA) was developed using a monoclonal antibody together with the EIAV p26 protein labelled with HRP. The test has been evaluated against the standard and with field serum samples globally. This bELISA test can be completed within 75 min, and the sensitivity is higher than those of either the AGID or one commercial cELISA kit. This bELISA assay was 8–16 times more analytically sensitive than AGID, and 2 to 4 times more analytically sensitive than one cELISA kit by testing three sera from the USA, Argentina, and China, respectively. The 353 serum samples from Argentina were tested, in comparison with AGID, the diagnostic sensitivity and specificity of our bELISA assay were 100% (154/154) and 97.0% (193/199), respectively, and the accuracy of the bELISA test was 98.3%. The bELISA test developed in this study is a rapid, sensitive, specific method for the detection of EIAV infection, and could be a promising candidate for use in the monitoring of the EIA epidemic worldwide. • A universal epitope-based blocking enzyme-linked immunosorbent assay (bELISA) was developed for detection of antibodies to EIAV. • The bELISA assay can be used to test EIAV serum samples from different regions of the world including North America, South America, Europe, and Asia. • The bELISA assay was evaluated in three different international labs and showed a better performance than other commercial kits.

Epizootic equine lymphangitis (EEL) is a chronic fungal disease that affects equids. The causative agent is a dimorphic fungus called Histoplasma capsulatum var farciminosum. Histoplasmacapsulatum var farciminosum field strain 7 (D 2878/2023) isolated from the eye socket of an EEL Ethiopian horse was sub-cultured on four different solid media and incubated at 26°C and 37°C for 6 weeks. Details of growth morphology were recorded and shown in images during 6 weeks of incubation. Histoplasmacapsulatum var farciminosum grew best at 26°C on all four agars, but only on sheep blood agar at 37°C as small, white dry colonies.

Pandemics like SARS-Cov-2 very frequently have their origin in different animals and in particular herds of camels could be a source of zoonotic diseases. This study took advantage on a highly sensitive and adaptable method for the fast and reliable detection of viral antibodies in camels using low-cost equipment. Magnetic nanoparticles (MNP) have high variability in their functionalization with different peptides and proteins. We confirm that 3-aminopropyl triethoxysilane (APTES)-coated MNP could be functionalized with viral proteins. The protein loading could be confirmed by simple loading controls using FACS-analysis (p < 0.05). Complementary combination of antigen and antibody yields in a significant signal increase could be proven by both FACS and COMPASS. However, COMPASS needs only a few seconds for the measurement. In COMPASS, the phase φn on selected critical point of the fifth higher harmonic (n = 5th). Here, positive sera display highly significant signal increase over the control or negative sera. Furthermore, a clear distinction could be made in antibody detection as an immune response to closely related viruses (SARS-CoV2 and MERS). Using modified MNPs along with COMPASS offers a fast and reliable method that is less cost intensive than current technologies and offers the possibility to be quickly adapted in case of new occurring viral infections. KEY POINTS: • COMPASS (critical offset magnetic particle spectroscopy) allows the fast detection of antibodies. • Magnetic nanoparticles can be adapted by exchange of the linked bait molecule. • Antibodies could be detected in camel sera without washing steps within seconds.

We developed an ELISPOT assay for evaluating Middle East respiratory syndrome coronavirus (MERS-CoV)‒specific T-cell responses in dromedary camels. After single modified vaccinia virus Ankara-MERS-S vaccination, seropositive camels showed increased levels of MERS-CoV‒specific T cells and antibodies, indicating suitability of camel vaccinations in disease-endemic areas as a promising approach to control infection.

Adult camel leukosis is an emerging hematological and neoplastic disease in dromedaries. It has been hypothesized that bovine leukemia virus (BLV) or its genetic variants may be associated with adult camel leukosis. In this study, we used next-generation sequencing (NGS) to detect all possible viruses in five lung samples from five dromedaries with histopathological evidence of adult camel leukosis and four tissue samples from two control dromedaries. A total throughput of 114.7 Gb was achieved, with an average of 12.7 Gb/sample. For each sample, all the pair-end 151-bp reads were filtered to remove rRNA sequences, bacterial genomes and redundant sequences, resulting in 1–7 Gb clean reads, of which <3% matched to viruses. The largest portion of these viral sequences was composed of bacterial phages. About 100–300 reads in each sample matched “multiple sclerosis-associated retrovirus”, but manual analysis showed that they were only repetitive sequences commonly present in mammalian genomes. All viral reads were also extracted for analysis, confirming that no BLV or its genetic variants or any other virus was detected in the nine tissue samples. NGS is not only useful for detecting microorganisms associated with infectious diseases, but also important for excluding an infective cause in scenarios where such a possibility is suspected.