Limited Genetic Diversity Detected in Middle East Respiratory Syndrome-Related Coronavirus Variants Circulating in Dromedary Camels in Jordan.

Middle East respiratory syndrome-related coronavirus (MERS-CoV) is a persistent zoonotic pathogen with frequent spillover from dromedary camels to humans in the Arabian Peninsula, resulting in limited outbreaks of MERS with a high case-fatality rate. Full genome sequence data from camel-derived MERS-CoV variants show diverse lineages circulating in domestic camels with frequent recombination. More than 90% of the available full MERS-CoV genome sequences derived from camels are from just two countries, the Kingdom of Saudi Arabia (KSA) and United Arab Emirates (UAE). In this study, we employ a novel method to amplify and sequence the partial MERS-CoV genome with high sensitivity from nasal swabs of infected camels. We recovered more than 99% of the MERS-CoV genome from field-collected samples with greater than 500 TCID50 equivalent per nasal swab from camel herds sampled in Jordan in May 2016. Our subsequent analyses of 14 camel-derived MERS-CoV genomes show a striking lack of genetic diversity circulating in Jordan camels relative to MERS-CoV genome sequences derived from large camel markets in KSA and UAE. The low genetic diversity detected in Jordan camels during our study is consistent with a lack of endemic circulation in these camel herds and reflective of data from MERS outbreaks in humans dominated by nosocomial transmission following a single introduction as reported during the 2015 MERS outbreak in South Korea. Our data suggest transmission of MERS-CoV among two camel herds in Jordan in 2016 following a single introduction event.

Seroprevalence and risk factors of Leptospira serovar Pomona and Leptospira serovar Hardjo infection in dairy cows in Jordan.

INTRODUCTION: This study was carried out to determine the seroprevalence and risk factors associated with Leptospira serovars Pomona and Hardjo infection in dairy cows. METHODOLOGY: Seroprevalence was determined using ELISA using 160 healthy and 80 recently aborted cows. Risk factor assessment was carried out using a pre-validated questionnaire. RESULTS: The true farm seroprevalence of Leptospira serovars Pomona and Hardjo was 92.3% (95% CI: 66%-98%). In healthy cows, the true and apparent cow seroprevalence of Leptospira serovars Pomona and Hardjo were 26.9 (95% CI: 20-34%), 26.25% (95% CI: 20-33%) and 28.75% (95% CI: 22-36%) and 27.5% (95% CI: 21-35%), respectively. Semi-intensive management system (OR = 11.43; P < 0.01), surface water as a source of drinking water (OR = 1.21; P < 0.03), lack of special wear for visitors (OR = 1.39; P < 0.05), and previous history of abortion (OR = 1.02; P < 0.05) were associated with high rate of seropositivity against Leptospira serovars Pomona and Hardjo. In recently aborted cows, the true and apparent seroprevalence rates of Leptospira serovars Pomona and Hardjo were 53.25% (95% CI: 47.5-62%), 53.75% (95% CI: 48.5-63.2%) and 56% (95% CI: 49-61%), 56.25% (95% CI: 49.8-61.2%), respectively. CONCLUSIONS: Leptospirosis is an endemic disease in Jordan and further studies are required to effectively control the disease in dairy cows.

High Prevalence of Middle East Respiratory Coronavirus in Young Dromedary Camels in Jordan.

Prevalence of Middle East respiratory syndrome coronavirus (MERS-CoV) was determined in 45 dromedary camels from two geographically separated herds in Jordan. Virus shedding was only detected in swabs obtained from the respiratory tract and primarily observed in camels younger than 3 years. MERS-CoV seroprevalence increased with age of camels. Bovine and sheep sera were seronegative. Phylogenetic analysis of partial S2 clustered the Jordanian MERS-CoV strains with contemporary MERS-CoV strains associated with nosocomial outbreaks.

Epidemiologic study on Besnoitia besnoiti infection in dairy herds in Jordan.

Besnoitia besnoiti is an apicomplexan parasite and the causative agent of bovine besnoitiosis which is considered as a re-emergent disease in Europe. A cross-sectional serological study was conducted to determine the seroprevalence and to identify risk factors associated with B. besnoiti infection in 68 dairy herds (n = 806 cows) in Jordan during the period from January to June 2007 and the spring of 2014. Data regarding herd's management was obtained by filling questionnaires through personal interviews with farmers. An indirect ELISA test was used to detect antibodies against B. besnoiti. Chi-square analysis and multivariable logistic regression model were used to identify risk factors associated with seropositivity to B. besnoiti. At the individual cow and herd level, the true prevalence of seropositive animals was 6 and 28.7 %, respectively. Cows between 2 and 6 years of age had significantly higher seroprevalence of B. besnoiti than other age groups. The highest seroprevalence of B. besnoiti was found in Zarqa and Irbid governorates. Multivariable logistic regression model identified that exchanging visits by farm workers to neighboring farms as a risk factor for seropositivity to B. besnoiti, while smaller herd size and twice a day farm cleaning using sweeping and water hosing were identified as protective factors. This is the first study that investigated the seroprevalence of B. besnoiti infection in dairy herds in Jordan. Further studies are warranted to explore the clinical manifestation of B. besnoiti infection as well as to identify the possible presence of other Besnoitia species and definitive hosts for the parasite.

Evaluation of the use of atropine sulfate, a combination of butylscopolammonium bromide and metamizole sodium, and flunixin meglumine to ameliorate clinical adverse effects of imidocarb dipropionate in horses.

OBJECTIVE: To evaluate the ability of atropine sulfate, butylscopolammonium bromide combined with metamizole sodium, and flunixin meglumine to ameliorate the clinical adverse effects of imidocarb dipropionate in horses. ANIMALS: 28 horses with piroplasmosis. PROCEDURES: 28 horses were randomly assigned to 4 equal groups according to the pretreatment administered. Fifteen minutes before administration of 2.4 mg of imidocarb dipropionate/kg IM, horses in the first group were pretreated with 0.02 mg of atropine sulfate/kg IV, the second group with a combination of 0.2 mg of butylscopolammonium bromide/kg IV and 25 mg of metamizole sodium/kg IV, the third group with 1.1 mg of flunixin meglumine/kg IV, and the fourth (control) group with 1 mL of saline (0.9% NaCl) solution/50 kg IV. Physical examination, including evaluation of rectal temperature, heart and respiratory rates, capillary refill time, mucous membrane color, hydration status, abdominal sounds, signs of abdominal pain, salivation, diarrhea, and number of defecations, was performed. RESULTS: Imidocarb dipropionate use in the control group was associated with serious adverse effects including signs of abdominal pain (4/7 horses) and diarrhea (2/7). Horses pretreated with atropine had no diarrhea, but 6 had signs of abdominal pain. Only 1 horse that received butylscopolammonium-metamizole pretreatment had signs of abdominal pain and 3 had diarrhea, which was numerically but not significantly different than the control group. Of horses pretreated with flunixin, 3 had signs of abdominal pain and 3 had diarrhea. CONCLUSIONS AND CLINICAL RELEVANCE: A combination of butylscopolammonium bromide and metamizole sodium may be useful to ameliorate the adverse effects of imidocarb dipropionate in horses, although group size was small and significant differences from the control group were not found.

Prevalence and risk factors associated with Neospora caninum infection in dairy herds in Jordan.

A cross-sectional study was carried out to determine the seroprevalence and to identify risk factors associated with Neospora caninum infection in 62 dairy herds (n = 671 cows) in Jordan between January and June 2007. Information regarding herd management was recorded through personal interviews with farmers. Antibodies against N. caninum were detected using an indirect ELISA test. Chi-square analysis and multivariable logistic regression model were used to identify risk factors associated with N. caninum seropositivity. The true prevalence of antibodies against N. caninum in individual cows and cattle herds was 35 and 66.5 %, respectively. There was no significant difference in the percentage of seropositive animals between different age groups. Multivariable logistic regression model revealed workers frequently visiting nearby farms as a risk factor for seropositivity to N. caninum, while presence of a calving pen was suggested as a protective factor. Amman, Balqa, and Mafrak governorates had significantly lower seroprevalence to N. caninum compared to other Jordanian governorates. Results of this study indicated that N. caninum infection may be widespread in Jordan.

Prevalence and risk factors associated with Chlamydophila abortus infection in dairy herds in Jordan.

A cross-sectional study was carried out to determine seroprevalence and to identify risk factors associated with Chlamydophila abortus infection in 62 nonvaccinated dairy herds (671 cows) in Jordan between January and June 2007. Information regarding herd management was recorded through a personal interview with farmers. Antibodies against C. abortus were detected using an ELISA test kit. Chi-square analysis and multivariable logistic regression model were used to identify risk factors associated with C. abortus seropositivity. The true prevalence of antibodies against C. abortus in individual cows and cattle herds were 19.9 % and 66.3 %, respectively. Univariable Chi-square analysis revealed three variables with P ≤ 0.25 that were further offered to multivariable logistic regression analysis. Small-sized herds were identified as a risk factor for seropositivity to C. abortus, while sweeping followed by water hosing and using disinfectants were identified as protective factors. Cows in the age groups of >8 and ≤ 10 years old and >2 and ≤ 6 years old had the highest and lowest significant seroprevalence to C. abortus, respectively. Results of this study indicated that C. abortus is highly prevalent in Jordan's dairy herds and Chlamydophila infection could be controlled by applying strict biosecurity measures in the dairy farms.

Distribution and characterization of the Escherichia coli heat-stable enterotoxin (STa) receptor throughout the intestinal tract of newborn camels (Camelus dromedaries).

Heat-stable enterotoxin (STa) secretion from Enterotoxigenic Escherichia coli (ETEC) is crucial for the pathogenesis of diarrhea in both animal and human. The goal of this study was to investigate the distribution of the STa-specific receptors in the newborn camel's enterocytes and brush border membrane vesicles (BBMVs). Flow cytometric analysis was used to investigate the density of STa-receptors on enterocytes and BBMVs prepared from anterior jejunum, posterior jejunum, ileum, and colon. Strong density and affinity of STa-receptors was present on enterocytes and BBMVs of the ileum compared to that in the other intestinal segments. It was concluded that the ileum is the major target for STa action in newborn camels.

Seroprevalence and risk factors for bovine brucellosis in Jordan.

We investigated the seroprevalence and risk factors for Brucella seropositivity in cattle in Jordan. The sera from 671 cows were randomly collected from 62 herds. The antibodies against Brucella were detected using a Rose Bengal plate test and indirect ELISA. A structured questionnaire was used to collect information on the cattle herds' health and management. A multiple logistic regression model was constructed to identify the risk factors for Brucella seropositivity. The true prevalence of antibodies against Brucella in individual cows and cattle herds was 6.5% and 23%, respectively. The seroprevalence of brucellosis in cows older than 4 years of age was significantly higher than that in the younger cows. The seroprevalence of brucellosis in cows located in the Mafraq, Zarqa and Ma'an governorates was significantly higher than that of the other studied governorates. The multiple logistic regression model revealed that a larger herd size (odd ratio OR = 1.3; 95% CI: 1.1, 2.6) and mixed farming (OR = 2.0; 95% CI: 1.7, 3.7) were risk factors for cattle seropositivity to Brucella antigens. On the other hand, the use of disinfectants (OR = 1.9; 95% CI: 1.1, 2.1) and the presence of adequate veterinary services (OR = 1.6; 95% CI: 1.2, 3.2) were identified as protective factors.

Childhood brucellosis in Jordan: prevalence and analysis of risk factors.

OBJECTIVES: To investigate the seroprevalence of childhood brucellosis in Jordan and to elucidate risk factors associated with seropositivity to Brucella. METHODS: Between January 2001 and October 2006, a cross-sectional study of children aged < or = 15 years was conducted; a total of 1282 serum samples were collected. Serum samples were analyzed for the presence of antibodies to Brucella antigens using the Rose Bengal plate test (RBPT) and a direct enzyme-linked immunosorbent assay (ELISA). Risk factors associated with seropositivity to Brucella antigens were identified by constructing a multivariate logistic regression model. RESULTS: Of the 1282 serum samples tested, 119 (9.3%) were positive by both RBPT and ELISA tests. When adjusted for the sensitivities and specificities of the two tests, the true seroprevalence was 11.6%. The seroprevalence of Brucella antibodies in Ma'an and Mafraq governorates was significantly higher than in the other governorates. The logistic regression model identified male gender (OR 2.5, 95% CI 1.4, 4.1), age older than 10 years (OR 1.8, 95% CI 1.1, 3.9), living in a village (OR 2.0, 95% CI 1.3, 2.9), and assisting in raising small ruminants (OR 1.6, 95% CI 1.1, 2.6) as risk factors for childhood Brucella seropositivity in Jordan. CONCLUSION: This study documents the importance of brucellosis in children and further emphasizes the role of small ruminants as an important source for human infection.

Risk factors associated with camel brucellosis in Jordan.

During the period between February, 2004 and December, 2006, a cross-sectional study was performed to investigate some epidemiological aspects related to camel brucellosis in Jordan. Four hundred twelve camel sera from 37 herds were randomly collected and analyzed using Rose Bengal plate test and complement fixation test. A structured pre-tested questionnaire was administered to collect information on camel herd health and management. A multivariable logistic regression model was constructed to investigate risk factors associated with seropositivity to Brucella antigens. Moreover, the incidence of Brucella-specific abortion was investigated in 7 camel herds located in different locations in Southern Jordan. The true prevalence of Brucella-seropositive in camels was 12.1%. Thirteen (35.1%) herds had at least one positive camel. The seroprevalence ofbrucellosis in camels was significantly higher in the southern part of Jordan than that in central or northern Jordan. The multivariable logistic regression model on both individual and herd levels revealed large herds and contact with small ruminants as risk factors for Brucella seropositivity. On the other hand, using disinfectants was identified as a protective factor (OR = 0.8; 95% CI: 0.1, 0.9) only on the camel herd level. The incidence of Brucella-caused abortion was 1.9%. Brucella melitensis biotype 3 was isolated from 4 aborted camel fetuses.

Seroprevalence of, and risk factors for, peste des petits ruminants in sheep and goats in Northern Jordan.

Peste des petits ruminants (PPR) is an economically important disease that affect sheep and goat industry in Asia and Africa. In this study, we investigated the seroprevalence, and risk factors, of PPR in sheep and goat flocks from five different governorates (Irbid, Jarash, Ajloun, Mafraq and Zarka) located in Northern Jordan. Serum samples from 929 and 400 sheep and goats, respectively, corresponding to 122 sheep flock and 60 goats flock were collected. Seroprevalence was determined using PPR competitive ELISA. Health status and management information were collected using a semi-structured pre-tested questionnaire. The individual true prevalence of PPR in sheep and goats was 29 and 49%, respectively. The flock level true prevalence of PPR was 60 and 74% in sheep and goats, respectively. In both sheep and goat flocks, large flock size, visiting live animals market and inadequate veterinary services were identified as risk factors for PPR seropositivity. Mixed (sheep and goats) raising was identified as a risk factor for PPR seropositivity in sheep flocks only.

Interaction of Escherichia coli heat-stable enterotoxin (STa) with its putative receptor on the intestinal tract of newborn kids.

The elaboration of heat stable enterotoxin (STa) is an important step in the pathogenesis of enterotoxigenic Escherichia coli (ETEC), which causes severe diarrhea in newborn animals. In this study, the distribution of the STa-specific receptors on enterocytes and brush border membrane vesicles (BBMVs) prepared from the anterior jejunum, posterior jejunum, ileum and colon of newborn kids was investigated. The density of STa-receptors on enterocytes and BBMVs was higher in the posterior jejunum than that in other segments of the kids' intestines. Additionally, the affinity of the posterior jejunum STa-receptors was higher than the affinity of receptors present on the epithelium of other intestinal segments. Our findings suggest that the posterior jejunum is a major target for STa within the intestinal tract of newborn kids.

Seroprevalence of Bartonella henselae and Bartonella quintana infections in children from Central and Northern Jordan.

OBJECTIVE: To investigate the prevalence of antibodies to Bartonella henselae (B.henselae) and Bartonella quintana (B.quintana) among children from central and northern Jordan. METHODS: Sera from 482 children were randomly collected from referenced governmental hospitals in the central and northern parts of Jordan during the period between January 2001 to March 2003. An indirect immunofluorescent assay was used to determine serum antibody titers to B.henselae and B.quintana. Sera that were reactive at a dilution >/-1:64 were considered positive. In addition, blood from 20 cats belonged to children with high B.henselae titers were tested using polymerase chain reaction. RESULTS: Out of the 482 serum samples examined, 53 (11%) and 20 (4.1%) had positive antibody titers for B. henselae and B.quintana. Bartonella henselae was isolated from 4 cats that belonged to 3 children with high antibody titers to B.henselae-IgG. The seroprevalence of IgG antibodies to B.henselae was significantly higher (P<0.05) in children aged 7-10-years than in younger or older ones. Having a cat in the household and having a history of cat scratches or bites were strongly associated (P<0.01) with seropositivity to B.henselae-specific IgG. Cats ownership and history of cat scratches or bites had no impact on the prevalence of B.quintana. Seropositivity to B.henselae-specific IgG was significantly higher (P<0.01) in children from northern Jordan than in children from central Jordan. CONCLUSION: This study substantiates the presence of B.henselae in Jordan, documents the seroreactivity to 2 Bartonella antigens, and suggests that cat ownership and history of cat scratches or bites are important epidemiological risk factors for B.henselae infection in Jordan.

Seroprevalence of and risk factors for Bartonella henselae and Bartonella quintana infections among pet cats in Jordan.

To determine the seroprevalences of Bartonella henselae and Bartonella quintana antibodies in Jordanian pet cats, serum samples from 153 cats from three geographical regions were analyzed. Seroprevalences were determined by indirect immunofluorescence. The true seroprevalences to B. henselae and B. quintana were 32 and 1.5%, respectively. The seroprevalence of B. henselae-specific antibodies in cats from northern Jordan was significantly higher than in cats from the central or southern parts of Jordan. The seroprevalence to B. henselae increased to age 2 years. Odds of seropositivity were higher in cats living outdoors, showing hunting behavior, having a flea infestation and of a mixed breed. No association was detected with sex.