Virološko i molekularno istraživanje na uzorcima bizona pod sumnjom da su zaraženi virusom bizonskih boginja u Egiptu 2019. godine

In this study, skin lesions from buffaloes showing clinical signs of buffalopox infection were tested to isolate and identify the buffalopox virus (BPXV). Clinical examination of infected buffaloes was performed and visible clinical signs recorded. Skin scabs from infected buffaloes were collected and used for virus isolation on embryonated chicken egg (ECE) and tissue culture cell lines. The isolated BPXV was identified and characterized using polymerase chain reaction (PCR). The infected buffaloes displayed fever, skin eruptions, enlargement of superficial lymph nodes, emaciation and drop in milk yield. The ECE inoculated with the prepared skin scab samples showed clear raised white pock lesions on the chorioallantoic membrane (CAM). The inoculated tissue cultures (VERO and BHK cell lines) revealed a cytopathic effect (CPE) including rounding, clumping with cytoplasmic granulation and cluster formation. PCR for the C18L specific BPXV gene was carried out on the virus infected tissue culture produced 368 bp bands. Human infection with BPXV was also recorded. It was concluded that BPXV is circulating in Egyptian buffaloes, causing economical losses and infection in contact humans.U ovoj studiji lezije kože bizona koje su pokazivale kliničke znakove infekcije bizonskim boginjama, rabljene su za izoliranje i identifikaciju virusa bizonskih boginja (BPXV). Provedeno je kliničko ispitivanje zaraženih bizona i zabilježeni su očiti klinički znaci. Kraste s kože zaraženih bizona su prikupljene i rabljene za izolaciju virusa na embrioniranom kokošjem jajetu (ECE) te linijama tkivnih staničnih kultura. Izolirani BPXV identificiran je i okarakteriziran uporabom lančane reakcije polimerazom (PCR). Zaraženi bizoni imali su vrućicu, osip na koži, povećanje površinskih limfnih čvorova, mršavjeli su, a i došlo je do pada proizvodnje mlijeka. ECE cijepljeni pripremljenim uzorcima krasta s kože pokazali su jasne uzdignute bijele lezije boginja na korioalantoičnoj membrani (CAM). Cijepljene tkivne kulture (VERO i BHK stanične linije) otkrile su citopatski učinak (CPE), uključujući zaobljivanje, agregaciju s granulacijom citoplazme i formiranje klastera. PCR za C18L specifični gen virusa bizonskih boginja proveden je na tkivnoj kulturi zaraženoj virusom proizvevši 368 bp vrpci. Zabilježena je i zaraza ljudi BPXV-om. Zaključeno je da BPXV kruži među egipatskim bizonima, što prouzroči, dođu li u kontakt s BPXV zarazu ljudi i velike ekonomske gubitke

Prevalence and Classification of Amphistomes in Cattle and Buffaloes

Amphistomes are snail-borne trematodes infect rumens and reticulums capable of causing acute and chronic disease in cattle and buffaloes. A total of 897 of cattle and buffaloes were examined by faecal examinations and by postmortem examinations in Giza and Garbia governourates. The collected Amphistomes were morphologically and histologically classified. We found that the incidence of Amphistomes in totally examined animals was 4.9%. The incidence was higher in the oldest animals(than young), in the spring (than other seasons) and in Garbia (than Giza). But the incidence was the same in males and females. The collected Amphistomes were classified as Paramphistomum microbothrium, Paramphistomum cervi and Carmyerius gergaerius. We concluded that Amphistomes are prevalent among the examined cattle and buffaloes in Giza and Garbia governorates.

Epidemiology, diagnosis and therapy of Theileria equi infection in Giza, Egypt

Aim: To study the prevalence of Theileria equi among horses in different age groups, both sexes, months and seasons of the year, and regions of Giza governourate. Studying the changes in the blood picture, blood chemistries, liver enzymes associate with T.equi infections in horses. Evaluating IFA and CFT at different dilutions in the serodiagnosis of T.equi infections in horses. Evaluating four anti-Theileria medication regimens (diminazine aceturate, imidiocarb 7%, buparvaquone and a combination of imidiocarb 7% and buparvaquone) in treatment of T.equi infections in horses. Materials and Methods: Total of 149 horses were examined by clinical signs and blood smears. Fortey whole blood samples from T.equi infected horses were examined to measure haemoglobin, total RBCs count and PCV. Fortey serum samples from T.equi infected horses were examined to measure total bilirubin, direct bilirubin, ALT and AST enzymes. Serum samples from T.equi infected (40) and non infected (14) horses were tested by indirect fluorescent antibody test (IFA) and complement fixation test (CFT) at different dilutions. Four groups of T.equiinfected horses (A,B,C,D), each group was represented by 10 horses and was separately treated with diminazine aceturate, imidiocarb 7%, buparvaquone and a combination of imidiocarb 7% and buparvaquone respectively. Results: the prevalence of T.equi was 41.61% in totally examined horses. The prevalence was higher in males than females. The highest prevalence was among age group ranged from 5-10 years as (22.81%). The highest prevalence was in July and was recorded as (25.81%) and the disease was more prevalent in summer than winter. The highest prevalence was recorded in Nazlet-alsamman as (51.61%). Equine theileriosis was clinically characterized by fever, haemoglobinuria, oedema, anaemia and icterus. The best dilution for IFA was 1/160 where sensitivity, specificity and accuracy were the highest for this test as (98%), (92.86%) and (97.44%) respectively. The best dilution for CFT was 1/32 where sensitivity, specificity and accuracy were collectively the best as (90%), (92.86%) and (90.74%) respectively. Conclusion: It was concluded that T.equi is prevalent among horses in Giza governourate, its prevalence is varied according to the age, sex of horses, months, seasons and regions. T.equiinfections in horses are accompanied with changes in blood pictures, blood chemistries and liver enzymes. Both IFA and CFT could be used for the serodiagnosis of T.equi . The used four anti-Theileria medication regimens have the same ability to eradicate T.equi from the infected horses. [Vet World 2013; 6(2.000): 76-82

Isolation and molecular characterization of Mycoplasma spp. in sheep and goats in Egypt

Background and Aim: Different species of Mycoplasma are associated with many pathological problems in small ruminants including respiratory manifestation, this problem results in significant losses, especially in African countries. This study aimed to (I) study some epidemiological aspects of Mycoplasma species infections in Egyptian sheep and goats at Giza Governorate, (II) diagnosis of Mycoplasma species affections using bacterial isolation and identification, (III) apply the polymerase chain reaction (PCR) for typing of different Mycoplasma species, and (IV) illustrate the phylogenetic tree for the isolated Mycoplasma species and other species from GenBank using the purified PCR product. Materials and Methods: A total of 335 samples were collected from sheep and goats from Giza Governorate in Egypt as 142 nasal swabs from clinically affected animals, 167 pneumonic lungs, 18 samples from tracheal bifurcation, and 8 samples by bronchial wash were cultured on pleuropneumonia-like organisms (PPLOs) media for cultivation of Mycoplasma species. PCR and sequencing and phylogenetic analysis were adopted to identify and classify the isolated Mycoplasma species. Results: A total of 24 Mycoplasma isolates were isolated on PPLO media, identified by biochemical tests, and confirmed and typed by PCR using specific primers. 10 isolates were confirmed as Mycoplasma arginini, four isolates as Mycoplasma ovipneumoniae by PCR, and 10 isolates as undifferentiated Mycoplasma species. A purified isolate of M. arginini and M. ovipneumoniae was sequenced and phylogenetic analysis was illustrated. Conclusion: M. arginini and M. ovipneumoniae are prevalent in Egyptian sheep and goats. Further studies on M. arginini are required due to its high frequency of isolation from pneumonic sheep and goats and also from animals suffer from different respiratory manifestations

Morphological and molecular identification of the brown dog tick Rhipicephalus sanguineus and the camel tick Hyalomma dromedarii (Acari: Ixodidae) vectors of Rickettsioses in Egypt

Aim: Rickettsioses have an epidemiological importance that includes pathogens, vectors, and hosts. The dog tick Rhipicephalus sanguineus and the camel tick Hyalomma dromedarii play important roles as vectors and reservoirs of Rickettsiae. The aim of this study was to determine the prevalence of Rickettsiae in ixodid ticks species infesting dogs and camels in Egypt, in addition to, the morphological and molecular identification of R. sanguineus and H. dromedarii. Materials and Methods: A total of 601 and 104 of ticks’ specimens were collected from dogs and camels, respectively, in Cairo, Giza and Sinai provinces. Hemolymph staining technique and OmpA and gltA genes amplification were performed to estimate the prevalence rate of Rickettsiae in ticks. For morphological identification of tick species, light microscope (LM) and scanning electron microscope (SEM) were used. In addition to the phylogenetic analyses of 18S rDNA, Second internal transcript spacer, 12S rDNA, cytochrome c oxidase subunit-1, and 16S rDNA were performed for molecular identification of two tick species. Results: The prevalence rate of Rickettsiae in ticks was 11.6% using hemolymph staining technique and 6.17% by OmpA and gltA genes amplification. Morphological identification revealed that 100% of dogs were infested by R. sanguineus while 91.9% of camels had been infested by H. dromedarii. The phylogenetic analyses of five DNA markers confirmed morphological identification by LM and SEM. The two tick species sequences analyses proved 96-100% sequences identities when compared with the reference data in Genbank records. Conclusion: The present studies confirm the suitability of mitochondrial DNA markers for reliable identification of ticks at both intra- and inter-species level over the nuclear ones. In addition to, the detection of Rickettsiae in both ticks’ species and establishment of the phylogenetic status of R. sanguineus and H. dromedarii would be useful in understanding the epidemiology of ticks and tick borne rickettsioses in Egypt