Mustafa M. Ababneh

Middle East Respiratory Syndrome coronavirus (MERS- CoV) serology in major livestock species in an affected region in Jordan, June to September 2013

Between June and September 2013, sera from 11 dromedary camels, 150 goats, 126 sheep and 91 cows were collected in Jordan, where the first human Middle-East respiratory syndrome (MERS) cluster appeared in 2012. All sera were tested for MERS-coronavirus (MERS-CoV) specific antibodies by protein microarray with confirmation by virus neutralisation. Neutralising antibodies were found in all camel sera while sera from goats and cattle tested negative. Although six sheep sera reacted with MERS-CoV antigen, neutralising antibodies were not detected.

Prevalence and risk factors associated with bovine viral diarrhea virus infection in dairy herds in Jordan

A cross-sectional study was carried out to determine the seroprevalence and to identify risk factors associated with bovine viral diarrhea virus (BVDV) infection in 62 non-vaccinated 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 BVDV were detected using an indirect ELISA test. Chi-square analysis and multivariable logistic regression model were used to identify risk factors for BVDV seropositivity. The true prevalence of antibodies against BVDV in individual cows and cattle herds was 31.6% and 80.7%, respectively. The seroprevalence of BVDV in medium and large size herds was significantly higher than that in smaller herds. There was no significant difference in BVD seroprevalence between different age groups. Random-effects logistic regression model revealed two major factors associated with seropositivity to BVDV; exchange of visits between adjacent farm workers and not isolating newly purchased animals before addition to the herd. The seroprevalence of BVDV in cows located in the northern Jordanian governorates was significantly higher than that in other studied governorates. Results of this study indicated that BVDV is highly prevalent in Jordan and BVDV infection could be controlled by livestock-trade control, and applying strict biosecurity measures in the dairy farms.

Thermal manipulation during chicken embryogenesis results in enhanced Hsp70 gene expression and the acquisition of thermotolerance

The aim of this study was to evaluate the effect of thermal manipulation (TM) during embryogenesis on hatchability, growth performance and thermotolerance acquisition parameters during thermal challenge (TC). Seven-hundred and fifty fertile chicken eggs were divided randomly into three groups (250 eggs each): control group was maintained at 37.8°C and 56% relative humidity (RH), TM1 was subjected to TM at 38.8°C for 6h and 65% RH during embryonic days (ED)10-18 and TM2 was subjected to TM at 38.8°C for 18 h and 65% RH during ED10-18. Hatched chicks from each treatment group were then randomly divided into two sub-treatment groups (Naive and TC). Chicks in TC groups were subjected to TC by adjusting room temperature to 41.0°C for 6h on days 3, 7, and 42 of age while naïve chicks were kept under regular conditions (25 ± 1°C and 50-60% RH). Percentage of hatched eggs was recorded and post-hatch chick performance was evaluated by recording chick body weight (BW). Chicks response to TC was evaluated by determination of body temperature (T(b)), plasma T3 and T4 levels, and muscle mRNA levels of Hsp70. There was a significant increase in muscle mRNA levels of Hsp70 during embryogenesis and during TC in post-hatch chicks. While hatchability was not adversely affected, the body weight in TM2 chicks was significantly higher at the end of the study period (42 days). Results of this study indicated a long-term enhancement of Hsp70 gene expression associated with improved thermotolerance acquisition in treated chicks without adversely affecting performance.

Presence of Infectious Bronchitis Virus Strain CK/CH/LDL/97I in the Middle East

Infectious bronchitis virus (IBV) is a very dynamic and evolving virus, causing major economic losses to the global poultry industry. In early 2011, respiratory disease outbreaks were investigated in Iraq, Jordan, and Saudi Arabia. Five IBV isolates (JOA2, JOA4, Saudi-1, Saudi-2, and Iraqi IBV) were detected by diagnostic-nested nucleocapsid RT-PCR. Strain identification was characterised by sequencing and phylogenetic analysis of the amplified hypervariable region of the spike 1 (S1) gene. These five IBV isolates were found to be of the IBV strain CK/CH/LDL/97I. Nucleotide identity between these five IBV isolates ranged from 96.9% to 99.7%, and between these isolates and the CK/CH/LDL/97I strain in the range of 96.6–99.1%. The sequenced fragment of the S1 gene of the CK/CH/LDL/97I strain had less than 80% nucleotide identity to the IBV vaccine strains commonly used in the Middle East (M41 and H120). The presence of these CK/CH/LDL/97I-like strains may account for vaccination failure against IBV, since all IBV isolates were from vaccinated chickens. In this paper, we documented for the first time the presence of IBV strain CK/CH/LDL/97I in the Middle East. This strain is known to have originated in China and Taiwan.

Lumpy Skin Disease in Jordan: Disease Emergence, Clinical Signs, Complications and Preliminary-associated Economic Losses.

The objectives of this study are to report the emergence of lumpy skin disease (LSD) in Jordan and associated clinical signs, complications and preliminary economic losses. In mid-April, 2013, two adult dairy cattle developed clinical signs suggestive of LSD and were confirmed as positive by PCR. The two cases were in Bani Kenanah district, Irbid governorate, on the Jordanian border of Israel and Syria. The disease spread rapidly to all the districts of Irbid governorate. During the month following the emergence of the disease, data were collected related to the epidemiology of the disease and the numbers of affected cattle on the premises. Forty-one dairy cattle holdings were surveyed. The morbidity rate ranged from 3% to 100%, (Mean = 35.1%, SD ±28.5%). The mortality rate ranged from 0% to 20%, (Mean = 1.3%, SD ±4.4%). The case fatality rate ranged from 0% to 100%, (Mean = 6.2%, SD ±22%). The overall morbidity rate was 26%, mortality rate 1.9% and case fatality rate 7.5%. Skin nodules, anorexia, decreased milk production and decreased body weight were common clinical signs, while mastitis and myiasis were seen as complications in a few affected animals. Decreased body weight ranged from 0% to 80%, (Mean = 23.1%, SD ±15.7%). Decreased milk production ranged from 0% to 100%, (Mean = 51.5%, SD ±22.2%). Affected cattle were treated mainly with broad-spectrum antibiotics and anti-inflammatory drugs. The cost of treatment ranged from 0 to 84.3 British Pound/animal, (Mean = 27.9 GBP, SD ±22.5 GBP). LSD continues to spread through the Middle East region and poses a serious threat to the rest of Asia and Europe. International collaboration and communication is warranted to prevent the further spread of the disease to the rest of Asia and Europe.

Ovine and caprine toxoplasmosis ( Toxoplasma gondii ) in aborted animals in Jordanian goat and sheep flocks

Two hundred and fifty-five biological samples (106 aborted foetal tissue samples and 149 blood samples from aborted sheep and goats) were collected from 188 animals during the lambing season from September 2009 to April 2010 from the Mafraq region of Jordan. The sampled animals belonged to 93 goat and sheep flocks that had cases of abortion. A total of 169 (66.3%) biological samples were collected from sheep and 86 (33.7%) from goats. Seventy-six (29.8%) biological samples (45 blood and 31 tissue samples) were positive for Toxoplasma gondii by PCR assay. The positive samples were obtained from 43 sheep and 23 goats. The overall toxoplasma-specific prevalence rate was 35.1% (66/188). Forty flocks (43%) had at least one T. gondii PCR-positive animal. The risk factors related to flock health status and farm management that are hypothesized to be associated with T. gondii PCR positivity were also assessed using multiple logistic regressions. The presence of cats (OR = 4.74), a large flock size (OR = 2.76) and the method of disposing the aborted foetuses (OR = 3.77) were all statistically significant (P < 0.05) risk factors that were positively associated with toxoplasma positivity in goat and sheep flocks.

Applying variations of the Ovsynch protocol at the middle of the estrus cycle on reproductive performance of lactating dairy cows during summer and winter.

Two modifications of the Ovsynch protocol, GnRH+TAI after PGF(2 alpha) 48 h (CO-48) or 72 h (CO-72), were compared with the original protocol (OVS: GnRH-7 d-PGF(2 alpha)-2 d-GnRH-16 h-TAI) to study their effects on reproductive performance in 785 lactating dairy cows (Holstein Friesian, Bos Taurus). Results showed that more cows (P<0.001) returned to estrus within a week after TAI with CO-48 treatment compared with that in OVS and CO-72 treatments. Pregnancy rates were greater (P<0.001) for the CO-72 cows than those for both OVS and CO-48 treatments and for primiparous cows compared with multiparous cows. Moreover, pregnancy rates were lower in summer compared with those in winter. Pregnancy losses for cows in both CO-48 and CO-72 were greater (P<0.05) than that for cows in OVS treatment. Pregnancy losses were greater in summer (P<0.001) than in winter and for multiparous cows (P<0.001) than for primiparous cows. In conclusion, primiparous and winter-bred cows had greater pregnancy rates and fewer pregnancy losses than those of multiparous cows and summer-bred cows, respectively. Because of the presence of significant treatment, parity, and season interactions, TAI with ovulation synchronization protocols should be tailored according to the season and parity. CO-72 is recommended for primiparous cows but not for multiparous cows, and CO-48 is not recommended for synchronization. Furthermore, cows that exhibited estrus at any time were inseminated to improve pregnancy rates in ovulation synchronization protocols.

Pathological, immunological and molecular diagnosis of rabies in clinically suspected animals of different species using four detection techniques in Jordan.

A total of 29 brain tissue samples (BTS) were examined for rabies infection by different diagnostic techniques. None of the examined brain tissues were presented as a whole intact brain. Twenty-seven brain tissue samples from various animal species - dog (13 cases), cat (one case), fox (one case), pig (one case), cow (three cases), sheep (two cases), goat (one case), camel (one case), horse (one case) and donkey (three cases) - were provided by the Vaccine and Sera Department/Al-Bashir Central Hospital in Amman/Jordan from July 2009 up to May 2010. All these samples were frozen at -20°C, for a period of time and then fixed in 10% formalin after being tested for rabies virus by fluorescence antibody test (FAT). The results showed that 21 (77.77%) of 27 BTS were positive for rabies by FAT. Seventeen samples (58.62%) of 29 were positive by histopathology, 2 (6.90%) were positive by histopathology, immunohistochemistry (IHC) and of those which were fixed for 24h only, and 21 (72.42%) were positive using RT-PCR assay. Five of 29 BTS had no pathological lesions, 17 had Negri bodies and the remaining had non-suppurative encephalitis and necrosis. Thirteen BTS that were diagnosed positive by FAT were also positive by RT-PCR and histopathology, but negative by IHC. Four BTS that were positive by FAT were negative by histopathology, IHC and RT-PCR. Also, 3 BTS (cases 19, 22, and 25) that were negative by FAT were positive by RT-PCR and negative by IHC. One of these was negative, while two were positive by histopathology. Therefore, definitive diagnosis of rabies under these conditions in Jordan needs one or more other diagnostic tests in addition to FAT. Also, freezing and prolonged formalin fixation of BTS is not suitable for the detection of rabies virus antigen using IHC.

Molecular characterization of a recent Newcastle disease virus outbreak in Jordan.

Newcastle disease (ND) is a highly contagious viral disease and is a continuous threat to the poultry industry worldwide. In the early months of 2011, several devastating ND outbreaks occurred in Jordan affecting broilers, layers and breeders. The fusion gene of the isolated Newcastle disease virus (NDV) was partially amplified by RT-PCR, then directly sequenced. The NDV isolates were found to have the motif112RRQKRF117. This motif and a mean death time (MDT) of 46 h are indicative of the velogenic nature of these NDV isolates. Phylogenetic analysis showed that the new NDV strain belongs to the lineage 5d (Aldous et al., 2003) and is closely related to the Chinese strain SG/Liaoning/2009. NDV outbreaks in 2010 and 2011 have been noted in neighboring countries. Based on the high nucleotide similarity between our isolated NDV isolates and the Chinese NDV strain, the origin of these recent NDV isolates might be from China.

Ovine and caprine brucellosis (Brucella melitensis).

Last Updated: July 29, 2009 Importance Caprine and ovine brucellosis, caused by the zoonotic bacterium Brucella melitensis, is an economically important cause of abortion in small ruminants. This infection causes significant losses from decreased productivity and lost trade in much of the developing world. B. melitensis is considered to be a re-emerging pathogen in the Middle East. It has been eradicated from some nations, but the cost of surveillance to remain B. melitensis-free is significant. B. melitensis is also an important human pathogen. In humans, brucellosis is a serious, debilitating and sometimes chronic disease that can affect a variety of organs. Most cases are the result of occupational exposure to infected animals, but infections can also occur from ingesting contaminated dairy products. In addition, B. melitensis could be used in a bioterrorist attack.