Maged El-Ashker

Molecular biological identification of Babesia, Theileria, and Anaplasma species in cattle in Egypt using PCR assays, gene sequence analysis and a novel DNA microarray.

In this preliminary study, a novel DNA microarray system was tested for the diagnosis of bovine piroplasmosis and anaplasmosis in comparison with microscopy and PCR assay results. In the Dakahlia Governorate, Egypt, 164 cattle were investigated for the presence of piroplasms and Anaplasma species. All investigated cattle were clinically examined. Blood samples were screened for the presence of blood parasites using microscopy and PCR assays. Seventy-one animals were acutely ill, whereas 93 were apparently healthy. In acutely ill cattle, Babesia/Theileria species (n=11) and Anaplasma marginale (n=10) were detected. Mixed infections with Babesia/Theileria spp. and A. marginale were present in two further cases. A. marginale infections were also detected in apparently healthy subjects (n=23). The results of PCR assays were confirmed by DNA sequencing. All samples that were positive by PCR for Babesia/Theileria spp. gave also positive results in the microarray analysis. The microarray chips identified Babesia bovis (n=12) and Babesia bigemina (n=2). Cattle with babesiosis were likely to have hemoglobinuria and nervous signs when compared to those with anaplasmosis that frequently had bloody feces. We conclude that clinical examination in combination with microscopy are still very useful in diagnosing acute cases of babesiosis and anaplasmosis, but a combination of molecular biological diagnostic assays will detect even asymptomatic carriers. In perspective, parallel detection of Babesia/Theileria spp. and A. marginale infections using a single microarray system will be a valuable improvement.

Optimization of a Fluorescence-Based Assay for Large-Scale Drug Screening against Babesia and Theileria Parasites

A rapid and accurate assay for evaluating antibabesial drugs on a large scale is required for the discovery of novel chemotherapeutic agents against Babesia parasites. In the current study, we evaluated the usefulness of a fluorescence-based assay for determining the efficacies of antibabesial compounds against bovine and equine hemoparasites in in vitro cultures. Three different hematocrits (HCTs; 2.5%, 5%, and 10%) were used without daily replacement of the medium. The results of a high-throughput screening assay revealed that the best HCT was 2.5% for bovine Babesia parasites and 5% for equine Babesia and Theileria parasites. The IC50 values of diminazene aceturate obtained by fluorescence and microscopy did not differ significantly. Likewise, the IC50 values of luteolin, pyronaridine tetraphosphate, nimbolide, gedunin, and enoxacin did not differ between the two methods. In conclusion, our fluorescence-based assay uses low HCT and does not require daily replacement of culture medium, making it highly suitable for in vitro large-scale drug screening against Babesia and Theileria parasites that infect cattle and horses.

Traumatic Reticuloperitonitis in Water Buffalo (Bubalus bubalis): Clinical Findings and the Associated Inflammatory Response

The present study was carried out to describe the clinical picture of traumatic reticuloperitonitis (TRP) in water buffalo (Bubalus bubalis) and to evaluate the inflammatory and immunologic responses for this clinical condition. Twenty-two buffalo with acute local TRP were monitored in our study. Additionally, 10 clinically healthy buffalo were randomly selected and served as controls. Acute local TRP was initially diagnosed by clinical examination and confirmed by ultrasonographic (USG) examination and/or necropsy findings. Blood samples were collected from all examined buffalo to measure the respective levels of tumor necrosis factor alpha (TNF-α), interleukin (IL)-1β, IL-6, IL-10 and interferon gamma (INF)-γ, serum amyloid A (SAA), C-reactive protein (CRP), haptoglobin (Hp), fibrinogen (Fb), and serum sialic acid (SSA). It was found that TNF-α, IL-1β, IL-6, IL-10, SAA, CRP, Hp, Fb, and SSA were significantly higher in buffalo with TRP than the controls. Our findings suggest that the examined immunologic variables were helpful in documenting the inflammatory response in buffalo with TRP. However, their diagnostic usefulness only becomes apparent when considered in tandem with the clinical findings for any given animal, its anamnesis, and a subsequent USG assessment. Due to the frequent complications of TRP, more accurate indicators of its occurrence and severity would be useful.

Staphylococci in cattle and buffaloes with mastitis in Dakahlia Governorate, Egypt

The aim of this study was to provide the first detailed insight into the population structure of Staphylococcus aureus in one modern dairy farm (Gamasa) and several household cows and buffaloes in Dakahlia Governorate, Egypt. Eight hundred seventy-two quarter milk samples of 218 dairy cattle and buffaloes with clinical and subclinical mastitis were investigated. Bacteria were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and staphylococci were further characterized by DNA sequencing of 16S rRNA genes and microarray analysis. Staphylococcus aureus was present in 5.6% of all collected samples, whereas methicillin-resistant S. aureus (MRSA) represented 24.5% of all identified S. aureus (12/49). Six clonal complexes (CC) of S. aureus were detected. Staphylococcus aureus CC398 (ST291/813)-MSSA (methicillin-susceptible S. aureus) was identified frequently in the Gamasa farm in addition to a few CC5-MRSA-V isolates. However, a small number of different isolates of S. aureus were found in household cattle and buffaloes harboring different CC. The presence of these genotypes of S. aureus in milk might indicate a public health hazard, because all of these CC have previously been isolated from human patients. Thus, a recommendation was given to the owner of the dairy farm to review the hygiene regimen on the farm. In perspective, further investigation regarding S. aureus screening of all lactating cows and personnel on the farm is warranted.

Q fever in cattle in some Egyptian Governorates: a preliminary study

BackgroundQ fever, caused by Coxiella burnetii, is a zoonosis with great public health significance and can cause financial losses to animal owners. The knowledge of the epidemiology of Q fever in Egypt is limited. Reports on this disease are scarce. In 2012 and 2013, we carried out this investigation to estimate the seroprevalence of antibodies to Coxiella burnetii in dairy cows of nine farms located in the lower Egyptian Governorates of Dakahlia, Damietta and Port Said. 1,194 blood sera were randomly collected from apparently healthy Holstein Friesian dairy cows. The collected sera were tested by ELISA for Coxiella burnetii antibodies.ResultsAll farms tested positive with seroprevalences ranging from 2.9 to 26.7% on farms with less than 200 animals and 9.8 to 20.0% in farms with more than 500 animals. 158 cows (13.2%) had anti-Coxiella antibodies.ConclusionQ fever may be enzootic in the cattle herds investigated in Damietta, Port Said, and Dakahlia Governorates of the Nile delta in both smaller and larger herds. There is a need for further research on the impact of Q fever on both veterinary and public health. The results of this study should trigger more detailed epidemiological studies in ruminants as well as investigations into the etiology of atypical pneumonia and fever of unknown origin in humans.

Seroprevalence of Rift Valley fever virus in livestock during inter-epidemic period in Egypt, 2014/15

BackgroundRift Valley fever virus (RVFV) caused several outbreaks throughout the African continent and the Arabian Peninsula posing significant threat to human and animal health. In Egypt the first and most important Rift Valley fever epidemic occurred during 1977/78 with a multitude of infected humans and huge economic losses in livestock. After this major outbreak, RVF epidemics re-occurred in irregular intervals between 1993 and 2003. Seroprevalence of anti-RVFV antibodies in livestock during inter-epidemic periods can be used for supporting the evaluation of the present risk exposure for animal and public health. A serosurvey was conducted during 2014/2015 in non-vaccinated livestock including camels, sheep, goats and buffalos in different areas of the Nile River Delta as well as the furthermost southeast of Egypt to investigate the presence of anti-RVFV antibodies for further evaluating of the risk exposure for animal and human health. All animals integrated in this study were born after the last Egyptian RVF epidemic in 2003 and sampled buffalos and small ruminants were not imported from other endemic countries.ResultsA total of 873 serum samples from apparently healthy animals from different host species (camels: n = 221; sheep: n = 438; goats: n = 26; buffalo: n = 188) were tested serologically using RVFV competition ELISA, virus neutralization test and/or an indirect immunofluorescence assay, depending on available serum volume. Sera were assessed positive when virus neutralization test alone or least two assays produced consistent positive results. The overall seroprevalence was 2.29% (95%CI: 1.51–3.07) ranging from 0% in goats, 0.46% in sheep (95%CI: 0.41–0.5), and 3.17% in camels (95%CI: 0.86–5.48) up to 5.85% in buffalos (95%CI: 2.75–8.95).ConclusionOur findings assume currently low level of circulating virus in the investigated areas and suggest minor indication for a new RVF epidemic. Further the results may indicate that during long inter-epidemic periods, maintenance of the virus occur in vectors and also most probably in buffaloes within cryptic cycle where sporadic, small and local epidemics may occur. Therefore, comprehensive and well-designed surveillance activities are urgently needed to detect first evidence for transition from endemic to epidemic cycle.

The role of acute phase cytokines in the recovery and disease progress of Theileria annulata-infected cattle

The present study was conducted to evaluate the cytokine response following natural infection of Theileria annulata in cattle. Initial survey included 173 crossbred cattle which were examined for the presence of Theileria piroplasms. The investigated cattle were clinically and parasitologically examined. Blood samples were collected from all examined cattle for microscopic examination, PCR assays (using primers of Theileria spp., Babesia spp., and T. annulata), and cytokines measurement. It was found that 38 cattle were positive for the presence of at least one species of Theileria; meanwhile Babesia piroplasms were not detected either by microscopy or PCR assay. When T. Annulata-specific primers were used, 33 gave positive results. Twenty-two out of 33 T. annulata-infected cattle were only included in this study together with contemporaneous controls (n = 10). According to the severity of clinical signs, T. annulata-infected cattle were categorized into two groups; group 1 included ten cattle with mild clinical signs and group 2 included 12 cattle with overt clinical signs. Biochemically, tumor necrosis factor alpha, interleukin (IL)-1β, IL-6, IL-12, haptoglobin, and Fb were significantly higher (p < 0.05) in diseased cattle compared to control and in cattle of group 2 compared to those in group 1, while interferon gamma showed no significant variations between the two groups. We conclude that measurements of the pro-inflammatory cytokines following T. annulata infection provide a valuable and quantitative assessment of the response to infection. It seems likely that the pro-inflammatory cytokines play an important role in the host response to T. annulata. Our findings suggest that the pro-inflammatory cytokines are suitable markers of inflammatory reactions in T. annulata-infected cattle.

Q Fever: A Re-Emerging Disease?

Q fever is a mainly airborne zoonosis with public health concern throughout the world caused by the highly contagious, obligate intracellular bacteria Coxiella burnetii. It is an important occupational zoonosis since its discovery in 1935; it has been shown to infect a wide range of hosts, including humans. Although Q fever is a disease closely related to occupations such as handling livestock, most of the previous studies concerned with general population. A recent outbreak in Europe reminds us that this is still a significant pathogen of concern, very transmissible with a very low infectious dose. For these reasons it has also featured regularly on various threat lists, as it may be considered to be used as a bio-weapon. Therefore, we reviewed the literatures on Q fever to highlight the epidemiologic, economic and public health impact of Q fever as a basis for designing effective control strategies.

Evaluation of the Inflammatory Reaction in Calves with Acute Ruminal Drinking

The present study was carried out to evaluate the inflammatory response during acute ruminal drinking in milkfed calves and to describe the correlation between production of acute phase cytokines and the occurrence of depression in this clinical condition. For this purpose, twenty calves with acute ruminal drinking as well as ten clinically normal calves were included in the study. Blood and ruminal fluid samples were collected from all examined calves. The blood samples were used to obtain a blood gas profile and for estimation of selected acute phase proteins and pro-inflammatory cytokines. A depression score was adopted to emphasize the relationship between the severity of the clinical signs and levels of acute phase cytokines. It was found that interleukin-1beta; interleukin-6, interferon gamma, c reactive protein, haptoglobin, and fibrinogen were significantly higher in diseased calves compared to the control group. There was also a positive correlation between the depression score and the examined acute phase cytokines. Our findings suggest that the acidity of the rumen Leads to inflammation of papillae with subsequent distinct inflammatory reaction. It seems that the pro-inflammatory cytokines play a role in the pathogenesis of acute ruminal drinking in milk-fed calves.

Gentamicin-induced Acute KidneyInjury in Equines is associated withMarked Acute Phase Response: AnExperimental Study on Donkey(Equus asinus)

Gentamicin-induced Acute Kidney Injury in Equines is associated with Marked Acute Phase Response: An Experimental Study on Donkey (Equus asinus) Recently, there is growing evidence suggesting that acute kidney injury (AKI) in human and laboratory animals is associated with an inflammatory response that could play a role in tissue damage. However, such link has not previously been addressed in equines. The present study was designed to evaluate the effect of gentamicin (GEN) administration on the development of AKI and systemic inflammatory response in equines using donkey as a model. GEN (10%) was administered intravenously in six donkeys at a dose of 20 mg kg-1 BW thrice daily for 14 consecutive days. Three other donkeys were randomly assigned to receive saline solution and served as controls. The donkeys were clinically and sonographically examined throughout the experimental period. Blood and urine (U) samples were simultaneously collected at day (D) 7, and D 14 of GEN administration. Renal specimens from all donkeys were collected at D 14 and processed for routine histopathological examination. AKI was confirmed by sonography, laboratory measurements, histopathology and immunohistochemistry.