Molecular characterization of some equine vector-borne diseases and associated arthropods in Egypt.

Equine vector-borne diseases (EVBDs) are emerging and re-emerging diseases, and most of them are zoonotic. This study aimed to investigate EVBDs in equines and associated arthropods (ticks and flies) from Egypt using molecular analyses, in addition to a preliminary characterization of associated ticks and flies by the matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) and molecular techniques. In this study, 335 blood samples were obtained from equines that appeared to be in good health (320 horses and 15 donkeys) in Cairo and Beni Suef provinces, Egypt. From the same animals, 166 arthropods (105 sucking flies and 61 ticks) were collected. Ticks and flies were preliminary characterized by the MALDI-TOF and molecular tools. Quantitative PCR (qPCR) and standard PCR coupled with sequencing were performed on the DNA of equines, ticks, and flies to screen multiple pathogens. The MALDI-TOF and molecular characterization of arthropods revealed that louse fly (Hippobosca equina) and cattle tick (Rhipicephalus annulatus) infesting equines. Anaplasma platys-like (1.6%), Anaplasma marginale (1.6%), Candidatus Ehrlichia rustica (6.6%), a new Ehrlichia sp. (4.9%), and Borrelia theileri (3.3%) were identified in R. annulatus. Anaplasma sp. and Borrelia sp. DNAs were only detected in H. equina by qPCR. A. marginale, Anaplasma ovis, and Theileria ovis recorded the same low infection rate (0.6%) in donkeys, while horses were found to be infected with Theileria equi and a new Theileria sp. Africa with recorded prevalence rates of 1.2% and 2.7%, respectively. In conclusion, different pathogens were first detected such as A. platys-like, Candidatus E. rustica, and a new Ehrlichia sp. in R. annulatus; A. marginale, A. ovis, and T. ovis in donkeys; and a new Theileria sp. "Africa" in horses.

Preparation and efficacy of freeze-dried inactivated vaccine against bovine viral diarrhea virus genotypes 1 and 2, bovine herpes virus type 1.1, bovine parainfluenza-3 virus, and bovine respiratory syncytial virus.

PURPOSE: Bovine respiratory disease is a worldwide health concern in the feedlot cattle causing morbidity and mortality in young with major economic losses to the producer. Programs of vaccination are integral parts of preventive health programs. We aim to prepare and evaluate lyophilized combined inactivated viruses (bovine viral diarrhea virus [BVDV] genotypes 1 and 2, bovine herpes virus type 1.1 [BoHV-1.1], bovine parainfluenza-3 virus [BPI-3V], and bovine respiratory syncytial virus [BRSV]) vaccine using saponin as a solvent and adjuvant in cattle. MATERIALS AND METHODS: Lyophilized Pneumo-5 vaccine was formulated to include the inactivated BVDV genotypes 1 and 2, BoHV-1.1, BPI-3V, and BRSV. The saponin solution was used as an adjuvant and solvent. The prepared vaccines were adjusted to contain 1- and 1.5-mg saponin/dose. It was evaluated for its sterility, safety, and potency in mice and calves. The antibody titers in vaccinated calves were measured by virus neutralization test and enzyme-linked immunosorbent assay (ELISA). RESULTS: The Pneumo-5 vaccine was found to be free from any contaminants and safe in mice. Meanwhile, the vaccine showed safety in calves which inoculated intramuscularly with the double dose of the vaccines. The overall immune response reached its peak in the 2nd-month post-vaccination. The vaccine contained saponin 1.5 mg/dose reached its antibodies peak in the 4th-week post-vaccination. All groups of vaccinated calves with both concentrations of the saponin did not show statistical significance in antibody titers measured by serum neutralization test and/or ELISA. CONCLUSION: The prepared vaccine, namely Pneumo-5, and adjuvanted with either 1 or 1.5 mg/dose saponin was proved safe and potent for effectual protection of calves against BVDV genotypes 1 and 2, BoHV-1.1, BPI-3V, and BRSV.

Molecular characterization of the 2018 outbreak of lumpy skin disease in cattle in Upper Egypt.

BACKGROUND AND AIM: Lumpy skin disease (LSD), an infectious disease of cattle, is characterized by raised nodules on the skin. Although the morbidity rate of LSD is low, it has a considerable fatality rate. Despite the annual mass vaccination of livestock with sheep pox vaccine (Veterinary Serum and Vaccine Research Institute, Egypt) enforced by Egyptian authorities, the LSD virus (LSDV) continues to circulate almost every summer. The present study aimed to discover the cause of cows naturally infected with LSDV circulating in Upper Egypt during the summer of 2018 using polymerase chain reaction (PCR) assay and to analyze their phylogenetics against reference genome sequences. MATERIALS AND METHODS: We cultured LSDV in specific pathogen-free embryonated chicken eggs (SPF-ECE) and used conventional PCR to identify fusion and P32 genes, previously deposited in GenBank (MN694826, MN694827, and MN954664). Sequencing and phylogenetic analyses were performed on these two highly conserved viral genes. RESULTS: LSDV infection of SPF-ECE resulted in characteristic white pock lesions. PCR products were identified on 1.5% agarose gel after electrophoresis at the expected positions for the fusion and P32 genes at 472 and 587 bp, respectively. CONCLUSION: The present study revealed that the two viral genes were identified from the Beni Suef and Sohag Governorates in all clinical cases and confirmed the circulation of LSDV in this outbreak. After sequencing, these genes were identical to those of the LSDV that had been identified and recorded in GenBank for the past 3 years.

In Vitro Effects of Streptomyces tyrosinase on the Egg and Adult Worm of Toxocara vitulorum.

BACKGROUND: Several species of streptomycetes, saprophytic bacteria found widely distributed in soil, water and plants, produce bioactive compounds such as intra and extracellular hydrolases including lytic enzymes which reflecting on their importance in the biological control of insects and parasites. This study assessed the in vitro effects of Streptomyces tyrosinase, produced from Streptomyces spp. isolated from Egyptian soil, on animal-parasitic nematode Toxocara vitulorum, in terms of egg development and adult worm's cuticular structure, and as an alternative strategy to alleviate this infection. METHODS: This study was conducted at the National Research Centre, Egypt in 2018. Five different concentrations of tyrosinase, ranged from 1%-30% were tested against the development of T. vitulorum eggs. The concentration induced the highest inhibitory activity was tested against adult T. vitulorum cuticle, which is essential for the protective and nutritive functions. The results were compared with those observed in the egg development and worm cuticle following incubation in Streptomyces protease (as a reference enzyme). RESULTS: Compared to Streptomyces protease, higher inhibitory activity on T. vitulorum egg development and extreme cuticular alterations of the treated adult worms had been observed following 24 h exposure to Streptomyces tyrosinase. Once the cuticle had been damaged, the enzyme would be able to penetrate deeper into the internal tissues of the nematode and caused more widespread disruption. CONCLUSION: The current study could offer a promising bio-control agent, Streptomyces tyrosinase, against T. vitulorum alternative to the more expensive synthetic anthelmintics.

Assessment of Haemonchus contortus larval and adult somatic antigens in sero-diagnosis of haemonchosis in naturally infected sheep and goats.

The current work was carried out to evaluate the potency of larval and adult somatic Haemonchus contortus (H. contortus) antigens in detection of haemonchosis among sheep and goats using ELISA. Two hundred and forty-three fecal and blood samples were randomly collected from small ruminants (107 sheep and 136 goats) in Beni-Suef Governorate, Egypt, during the period from June to August 2018. The fecal analysis exhibited that 26.33% of the small ruminants were infected with gastrointestinal nematodes. The overall prevalence of H. contortus was reached 22.22% whereas it was 27.10% and 18.38% among sheep and goats, respectively. The current study elucidated that the larval antigen has claimed more superior diagnostic results compared to the adult somatic H. contortus antigen. The apparent overall sero-prevalence among small ruminants was reached 51.85%. Separately, it was 64.48% in sheep and 41.91% in goats. The larval antigen had proved 96.55% sensitivity and 47.43% specificity, for sheep serum samples. Meanwhile, sensitivity and specificity for goats' sera were 100% and 71.17%, respectively. Diagnostic efficacy of ELISA was recorded 60.74% in sheep and 76.47% in goats. This study deduced that the larval antigen has proved the priority and the potency for diagnosis of H. contortus infection. Moreover, haemonchosis is a prevalent disease among the examined sheep and goats.

Tegumental Effects of Methanolic Extract of Balanites aegyptiaca Fruits on Adult Paramphistomum microbothrium (Fischoeder 1901) under Laboratory Conditions.

BACKGROUND: Weak efficacy of different fasciolicidal compounds used for treatment of paramphistomosis has drawn the attention of many authors to alternative drugs. The purpose of this study was to assess, for the first time, the effect of the methanolic extract of Balanites aegyptiaca fruits (BAE) on adult Paramphistomum microbothrium. METHODS: The effect of BAE on adult P. microbothrium after 24 h incubating the parasites in RPMI 1640 culture medium containing 10, 50, 100 and 200 µg/ml BAE was determined by light and scanning electron microscopic studies. RESULTS: Differences in response to BAE action were concentration dependent.The major target organ that was highly affected was the tegument. Maximum anthelmintic activity was found with a dose of 200 µg/ml BAE, at which distinct damage to the whole body surface of the trematodes was very much distinct. Shape and structure of both suckers were deformed due to BAE. This damage would undoubtedly disrupt many of the physiological processes associated with the tegument. Besides, the damage of the tegumental folds of the acetabular region might disrupt its function in drawing the rumen wall tissue of the host into the acetabular cavity. CONCLUSION: the use of methanolic extract of B. aegyptiaca fruits offers a new dimension and potential for control of such a neglected infectious disease in ruminants, at a time when paramphistomosis has emerged as an important cause of productivity loss.

Epidemiology of infant burn in Eastern Saudi Arabia.

OBJECTIVES: To identify the epidemiology, pattern, outcome, and impact of infant burns in Eastern Saudi Arabia. METHODS: We conducted a retrospective review of admitted infants charts over 4 years (2008-2013) at the Burn Unit of King Fahad Hospital, Hofuf, Al-Ahsa, Saudi Arabia. The charts were reviewed for age, gender, etiology, site of injury, total body surface area (TBSA), depth of burn, hospital stay, and discharge status. RESULTS: The total number of admissions to the Burn Unit was 510 cases. Out of these cases, 84 were infants, constituting 16.5% of total admissions. Scald burn was the most common etiology affecting 73 infants (86.9%). The highest percentage of total body surface area was between 5-10%, which occurred in 41 infants (48.8%). The average hospital stay was 10 days. No infant mortality was reported during this period. CONCLUSION: The prevalence of burns among infants in our hospital is high, and preventive measures must be implemented to decrease the occurrence of burns in this age group.

Characterization of the recent outbreak of foot-and-mouth disease virus serotype SAT2 in Egypt.

An outbreak of foot-and-mouth disease (FMD) in Egypt affected approximately 40,000 cattle and water buffaloes and killed more than 4,600 animals during February-March 2012. To investigate the etiology of the 2012 outbreak, specimens were collected from six governorates and analyzed using universal primers to amplify the 5' untranslated region (UTR) by reverse-transcription polymerase chain reaction. Only FMDV-SAT2 was detected, with an overall detection rate of 80.3 %. Complete VP1- and leader-proteinase-coding sequences, obtained from three isolates from three different governorates, were compared with previously reported sequences. Phylogenetic analysis of these sequences indicated that the circulating viruses were homogeneous and were closely related to topotype VII. Importantly, the newly emerged viruses were genetically closely related to strains isolated from Saudi Arabia, Sudan, Eritrea and Cameroon between 2000 and 2010, suggesting the dominant nature of this virus and underscoring the need for worldwide intensive surveillance to minimize its devastating consequences.