Identification of a novel lineage of Crimean-Congo haemorrhagic fever virus in dromedary camels, United Arab Emirates.

Crimean-Congo haemorrhagic fever virus (CCHFV) is a tick-borne virus causing Crimean-Congo haemorrhagic fever (CCHF), a disease reported to have a high fatality rate in numerous countries. The virus is geographically widespread due to its vector, and numerous wild and domestic animals can develop asymptomatic infection. Serological and limited molecular evidence of CCHFV has previously been reported in Camelus dromedarius (the dromedary, or one-humped camel) in the United Arab Emirates (UAE). In this study, 238 camel samples were screened for CCHFV RNA where 16 camel samples were positive for CCHFV by RT-PCR. Analysis of full-length CCHFV genome sequences revealed a novel lineage in camels from the UAE, and potential reassortment of the M segment of the genome.

Risk Factors for MERS-CoV Seropositivity among Animal Market and Slaughterhouse Workers, Abu Dhabi, United Arab Emirates, 2014-2017.

Camel contact is a recognized risk factor for Middle East respiratory syndrome coronavirus (MERS-CoV) infection. Because specific camel exposures associated with MERS-CoV seropositivity are not fully understood, we investigated worker-camel interactions and MERS-CoV seroprevalence. We assessed worker seroprevalence in 2 slaughterhouses and 1 live-animal market in Abu Dhabi, United Arab Emirates, during 2014-2017 and administered an epidemiologic survey in 2016 and 2017. Across 3 sampling rounds during 2014-2017, we sampled 100-235 workers, and 6%-19% were seropositive for MERS-CoV at each sampling round. One (1.4%) of 70 seronegative workers tested at multiple rounds seroconverted. On multivariable analyses, working as a camel salesman, handling live camels or their waste, and having diabetes were associated with seropositivity among all workers, whereas handling live camels and either administering medications or cleaning equipment was associated with seropositivity among market workers. Characterization of high-risk exposures is critical for implementation of preventive measures.

A study on some reproductive disorders in dromedary camel herds in Saudi Arabia with special references to uterine infections and abortion.

Dromedary camels complaining from conception failure or abortion were investigated and their herders interviewed in Al Ahsa province, Kingdom of Saudi Arabia (KSA) during 2013 and 2015. The most important reproductive disorder according to the responders is uterine infection (60.2%) followed by obesity (22.3%) then physiological conditions (hormonal disturbances; 7.8%), adhesions (3.9%) and repeat breeders (2.9%). Of the camel herders, 78.6% reported previous occurrence of abortion in their herds and 46% reported abortion cases in the last season (2015/2016), while 21.4% reported no history of abortion. Most of the responders (97.1%) do not call a veterinarian for cases of abortion in their herds and 53.4% do not discard aborted materials. The majority of the herders (76.7%) deny that handling aborted materials or touching vaginal fluids can result in human infection, or replied they do not know. Uterine swab samples were collected and tested by PCR for seven potential pathogens and sera tested for antibodies against bovine viral diarrhea virus (BVDV) and Brucella. Five pathogens were identified in infected uterine samples, namely Coxiella burnetii (36%), Campylobacter spp. (27%), Brucella spp. (17%), Salmonella spp. (13%), and Chlamydia spp. (7%). Sero-prevalence of Brucella and BVDV was 8.2 and 29.1% in overall sera, respectively, and varies with regard to the region. The findings of the present study demonstrate that reproductive disorders dominated by uterine infections and abortions are widespread in dromedary camels in KSA.

MERS-CoV in Upper Respiratory Tract and Lungs of Dromedary Camels, Saudi Arabia, 2013-2014.

To assess the temporal dynamics of Middle East respiratory syndrome coronavirus (MERS-CoV) infection in dromedary camels, specimens were collected at 1-2 month intervals from 2 independent groups of animals during April 2013-May 2014 in Al-Ahsa Province, Saudi Arabia, and tested for MERS-CoV RNA by reverse transcription PCR. Of 96 live camels, 28 (29.2%) nasal swab samples were positive; of 91 camel carcasses, 56 (61.5%) lung tissue samples were positive. Positive samples were more commonly found among young animals (<4 years of age) than adults (>4 years of age). The proportions of positive samples varied by month for both groups; detection peaked during November 2013 and January 2014 and declined in March and May 2014. These findings further our understanding of MERS-CoV infection in dromedary camels and may help inform intervention strategies to reduce zoonotic infections.

Multiplex PCR for rapid diagnosis and differentiation of pox and pox-like diseases in dromedary Camels.

BACKGROUND: Pox and pox-like diseases of camels are a group of exanthematous skin conditions that have become increasingly important economically. Three distinct viruses may cause them: camelpox virus (CMLV), camel parapox virus (CPPV) and camelus dromedary papilloma virus (CdPV). These diseases are often difficult to differentiate based on clinical presentation in disease outbreaks. Molecular methods such as PCR targeting species-specific genes have been developed and used to identify these diseases, but not simultaneously in a single tube. Recently, multiplex PCR has gained reputation as a convenient diagnostic method with cost-and timesaving benefits. METHODS AND RESULTS: In the present communication, we describe the development, optimization and validation of a multiplex PCR assay able to detect simultaneously the genome of the three viruses in one single test allowing for rapid and efficient molecular diagnosis. The assay was developed based on the evaluation and combination of published and new primer sets and was validated with viral genomic DNA extracted from known virus strains (n = 14) and DNA extracted from homogenized clinical skin specimens (n = 86). The assay detects correctly the target pathogens by amplification of targeted genes, even in case of co-infection. The method showed high sensitivity, and the specificity was confirmed by PCR-product sequencing. CONCLUSION: This assay provide rapid, sensitive and specific method for identifying three important viruses in specimens collected from dromedary camels with varying clinical presentations.

Phylogenetic analysis of eight sudanese camel contagious ecthyma viruses based on B2L gene sequence.

BACKGROUND: Camel contagious ecthyma (CCE) is an important viral disease of camelids caused by a poxvirus of the genus parapoxvirus (PPV) of the family Poxviridae. The disease has been reported in west and east of the Sudan causing economical losses. However, the PPVs that cause the disease in camels of the Sudan have not yet subjected to genetic characterization. At present, the PPV that cause CCE cannot be properly classified because only few isolates that have been genetically analyzed. METHODS AND RESULTS: PCR was used to amplify the B2L gene of the PPV directly from clinical specimens collected from dromedary camels affected with contagious ecthyma in the Sudan between 1993 and 2013. PCR products were sequenced and subjected to genetic analysis. The results provided evidence for close relationships and genetic variation of the camel PPV (CPPV) represented by the circulation of both Pseudocowpox virus (PCPV) and Orf virus (ORFV) strains among dromedary camels in the Sudan. Based on the B2L gene sequence the available CPPV isolates can be divided into two genetic clades or lineages; the Asian lineage represented by isolates from Saudi Arabia, Bahrain and India and the African lineage comprising isolates from the Sudan. CONCLUSION: The camel parapoxvirus is genetically diverse involving predominantly viruses close to PCPV in addition to ORFVs, and can be divided into two genetically distant lineages. Based on sequences of the B2L gene it is not possible to suggest that the viruses that cause CCE form a monophylogenetic group or species within the PPV phylogeny.

More cell culture passaged Camelpox virus sequences found resembling those of vaccinia virus.

Background: Camelpox is the most infectious and economically important disease of camelids that causes significant morbidity and mortality rates. Several live attenuated vaccines against Camelpox virus (CMLV) are produced worldwide by passaging field isolates in cell culture. Sequence of a high passage Saudi isolate of CMLV was previously found closely resembled Vaccinia virus (VACV). Aim: To determine whether other high cell culture passage CMLV isolates are genetically resemble VACV and further to explore the possible mechanism of the resemblance. Methods: We performed polymerase chain reaction and DNA sequence analysis of A-type inclusion body protein (ATIP), L1R, and open reading frame (ORF) 185 genes on different cell culture passage levels of a field isolate, two high passage vaccines, wild-type, and reference strains of CMLV. Results: We demonstrate that additional two high passage attenuated vaccine candidate from Sudan and UAE likewise contain sequences resembling VACV more than CMLV. Furthermore, sequence analysis of the ATIP gene of selected virus passages in cell culture revealed that the shift to VACV-like occurred between passage 11 and 20 and up to the 10th passage the genome still resembles wild-type virus. This observation was further confirmed by recombination analysis which indicated recombination events at ATIP and ORF185 genes occurred at higher passages. Conclusion: We confirmed that the cell culture passage CMLV turns to resemble VACV after cell culture passage and concluded that the resemblance may not be a result of contamination or misidentification as previously thought but could be due to recombination events that occurred during the passage process.

Genome Sequencing of a Camelpox Vaccine Reveals Close Similarity to Modified Vaccinia virus Ankara (MVA).

Camelpox is a viral contagious disease of Old-World camelids sustained by Camelpox virus (CMLV). The disease is characterized by mild, local skin or severe systemic infections and may have a major economic impact due to significant losses in terms of morbidity and mortality, weight loss, and low milk yield. Prevention of camelpox is performed by vaccination. In this study, we investigated the composition of a CMLV-based, live-attenuated commercial vaccine using next-generation sequencing (NGS) technology. The results of this analysis revealed genomic sequences of Modified Vaccinia virus Ankara (MVA).

Performance of an Immunochromatographic Test (ICT) in Comparison to Some Commonly Used Serological Tests for the Diagnosis of Brucellosis in Dromedary Camels (Camelus dromedarius).

Serological tests may represent an essential tool for the diagnosis of camel brucellosis; however, concerns arise in the scientific community regarding the direct transposition from cattle and small ruminants without adequate validation. The present study was made to compare four serological tests for the diagnosis of brucellosis in dromedary camels (Camelus dromedarius). In terms of sensitivity, our results show that the Immunochromatographic Test (ICT) shows the higher value of sensitivity, 98.67% (95% Confidence Level (C.L): 94.36%-99.99%), followed by the Fluorescence Polarization Assay (FPA) with 95.05% (95% C.L: 88.23%-99.51%), then the Competitive Enzyme-Linked Immunosorbent Assay (c-ELISA) with 94.94% (95% C.L: 88.25%-99.45%) and, finally, the Rose Bengal Test (RBT) with 68.95% (95% C.L: 56.55%-80.69%), which is the only test showing a significantly lower sensitivity compared to the others. On the other hand, our study revealed no significant difference in terms of specificity between all the tests under study, with a range from 99.06% (95% C.L: 98.34%-99.64%) for the ICT to 99.92% (95% C.L: 99.64%-100%) for the RBT. The ICT was found to be comparable in terms of sensitivity and specificity with the most commonly used tests for camel brucellosis. The results of the present study are of paramount importance for designing surveillance and control measures for brucellosis in camel populations.

Papillomavirus Infection in Humans and Dromedary Camels in Eastern Sudan.

Cases of wart-like lesions in humans and dromedary camels occurred in eastern Sudan in 2015 were described. Involvement of papillomavirus (PV) in causing these cases was affirmed by PCR and immunoperoxidase test. Mostly, the lesions were observed on the skin of the chest and forearms in addition to lips and mandible. Sequence analysis revealed Camelus dromedarius PV types 1 and 2 genotypes as the causative genotypes. We also observed cases of wart-like lesions on hands and legs of two herders attending the infected camel herd. Partial genome sequencing revealed human PV type 2 in one of the two human samples providing no indications for interspecies transmission of camel PVs, yet provides, for the first time evidence of active circulation of this virus in eastern Sudan.

Human and Dromedary Camel Infection with Camelpox Virus in Eastern Sudan.

We provide evidence for the zoonotic nature of camelpox virus by reporting infections that involved dromedary camels and three camel herders in Showak area of eastern Sudan between September and December 2014. The skin lesions in the camel herders consisted of erythema, vesicles, and pustules that involved arms, hands, legs, back, and abdomen and resolved within less than 2 months with no human-to-human transmission. The diagnosis was achieved through molecular technique, virus isolation in cell culture, and partial genome sequencing.

Serum biochemical profile and molecular detection of pathogens in semen of infertile male dromedary camels (Camelus dromedarius).

Detection of pathogens in the semen of camels has not been completely elucidated. Therefore, the current study aimed to determine the association of some economically important pathogens with infertility in 94 male infertile camels through molecular detection and estimation of selected biochemical parameters in serum of these animals compared with a control non infected fertile animals (n=40). PCR analysis of semen samples of infertile camels indicated that, four potential pathogens namely Mycoplasma spp., Leptospira spp., Brucella melitensis, and Bovine viral diarrhea virus (BVDV) were detected in 50 semen samples of infertile camels whereas, 44 semen samples of infertile camels were free of pathogens and all tested semen samples were negative for bovine herpes virus 1, Salmonella spp. and Trypanosoma evansi. Single and mixed infection was detected in 88% and 12% of the infected semen samples, respectively. Mycoplasma spp., Leptospira spp., Brucella and Bovine viral diarrhea virus infection represented 66%, 27.2%, 4.5% and 2.3% of the single infected semen samples. Mycoplasma spp.+Leptospira spp. and Mycoplasma spp.+Brucella spp. were detected in 83.3% and 16.7% of mixed infected semen samples, respectively. Testosterone concentration decreased significantly in infertile infected camels compare to both control and infertile non infected animals that remained comparable. The current findings reported the molecular detection of mixed infection in camel semen for the first time. Mycoplasma spp. is the most widely recognized microorganism in the present study and together with Leptospira spp., Brucella spp. and Bovine viral diarrhea virus, might be associated with infertility in dromedary camels.

Growth characteristic of Camel pox and Vaccinia viruses in embryonated eggs and cell culture.

Camel pox viruses isolated in Sudan and VD45 (African camel pox strain) and Vaccinia virus (Elstree strain) were used for inoculation of chorioallantoic membrane (CAM) of embryonated eggs (EE) and cell culture (CC). In EE Lesions were seen as pocks ranging in size from 1 to 1.5 mm in diameter, and they increase in size with serial passage and taking opaque- white and opaque- yellow colors. When propagated in Vero cells, these viruses gave clear CPE, characterized by rounding of cells, plaque formation, syncytia and detachment of cells from the glass.