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    Document

Characterization of Avian Influenza and Newcastle Disease Viruses from Poultry in Libya

On March 2013, the Libyan poultry industry faced severe outbreaks due to mixed infections of APMV-1 (Newcastle disease) and low pathogenic avian influenza (AI) of the H9N2 subtype which were causing high mortality and great economic losses. APMV-1 and H9N2 were isolated and characterized. Genetic sequencing of the APMV-1/chicken/Libya/13VIR/7225-1/2013 isolate revealed the presence of a velogenic APMV-1 belonging to lineage 5 (GRRRQKR*F Lin.5) or genotype VII in class II, according to the nomenclature in use. Three AI viruses of the H9N2 subtype, namely A/avian/Libya/13VIR7225-2/2013, A/avian/Libya/13VIR7225-3/2013, and A/avian/Libya/13VIR7225-5/2013, were isolated and found to belong to the G1 lineage. Analysis of amino acid sequences showed that the analyzed H9N2 viruses contained the amino acid Leu at position 226 (H3 numbering) at the receptor binding site of the HA, responsible for human virus-like receptor specificity. On March 2014, an outbreak of highly pathogenic avian influenza (HPAI) virus of the H5N1 subtype was diagnosed in a backyard poultry farm in an eastern region of Libya. The H5N1 isolate (A/chicken/Libya/14VIR2749-16/2014) was detected by real time RT-PCR (rRTPCR). Genetic characterization of the HA gene revealed that the identified subtype was highly pathogenic, belonged to the 2.2.1 lineage, and clustered with recent Egyptian viruses. This study revealed the presence of a velogenic APMV-1 genotype and of two influenza subtypes, namely HPAI H5N1 and H9N2, which are of major interest for public and animal health. Considering these findings, more investigations must be undertaken to establish and implement adequate influenza surveillance programs; this would allow better study of the epidemiology of APMV-1 genotype VII in Libya and evaluation of the current vaccination strategies.
Ibrahim Eldaghayes(5-2015)
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Isolation and molecular identification of Vibrio spp. by sequencing of 16S rDNA from seafood, meat and meat products in Libya

The genus Vibrio includes several food-borne pathogens that cause a spectrum of clinical conditions including septicemia, cholera and milder forms of gastroenteritis. Several Vibrio spp. are commonly associated with food-borne transmission including Vibrio cholerae, Vibrio parahemolyticus, and Vibrio vulnificus. Microbiological analysis for enumeration and isolation of Vibrio spp. were carried out for a total of 93 samples of seafood, meat and meat products from different geographic localities in Libya (Tripoli, Regdalin, Janzour and Tobruk). Vibrio spp. were detected by conventional cultural and molecular method using PCR and sequencing of 16S rDNA. Out of the 93 cultured samples only 48 (51.6%) yielded colonies on Thiosulfate Citrate Bile Salt agar (TCBS) with culture characteristics of Vibrio spp. More than half (n=27) of processed seafood samples (n=46) yielded colonies on TCBS, while only 44.6% of samples of meat and meat products showed colonies on TCBS. Among cultured seafood samples, the highest bacterial count was recorded in clam with a count of 3.8 х104 CFU\g. Chicken burger samples showed the highest bacterial count with 6.5 х104 CFU\g. Molecular analysis of the isolates obtained in this study, showed that 11 samples out of 48 (22.9%) were Vibrio spp. Vibrio parahemolyticus was isolated from camel meat for the first time. This study is an initial step to provide a baseline for future molecular research targeting Vibrio spp. foodborne illnesses. This data will be used to provide information on the magnitude of such pathogens in Libyan seafood, meat and meat products.
Ibrahim Eldaghayes(3-2016)
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Residual evaluation of oxytetracycline in camel edible tissues in Tripoli region, Libya

The deleterious effects of the residues of antibacterial drugs have been considered as one of the most serious problems in the world on the basis of their importance in both veterinary and human fields. Therefore, the present study was planned out to evaluate the residual levels of oxytetracycline in camel tissues in Tripoli area in Libya. Forty samples of slaughtered camel's tissues (10 of muscle, 10 of liver, 10 of kidney, and 10 of fat) were collected from different carcasses at different slaughter houses in Tripoli districts. The samples were homogenized, extracted and residual concentrations of oxytetracycline have been measured using liquid chromatography-Mass spectroscopy (LC-MS) technique and oxytetracycline standard. Oxytetracycline residues have been detected in 60% in muscle, 80% in liver, 90% in kidney and 70% in fat samples. The samples, although positive, yet decided acceptable as the detected levels were less than that were regulated by Codex Alimentarius Commission (CAC) for oxytetracycline maximal residual levels (100, 300, 600 and 100 μg/Kg muscle, liver, kidney or fat, respectively). The authors recommended avoiding irrational use of oxytetracycline in veterinary practice and camel in particular; and sticking to the withdrawal time regulated and labelled for drugs used in therapy among veterinary personnel, organizations, and governmental agencies in Libya.
Ibrahim Eldaghayes(9-2015)

Caprine Coccidiosis: An outbreak in the Green Mountain in Libya

This study involved a herd consisting of 200 goats in Green Mountain area, suffering from decrease in weight gain, bloody diarrhoea and severe anaemia and in some cases death within few days. Generally, there was no response to the treatment with antibiotics, anthelmentics and multivitamins. The last animal that died was submitted for post-mortem examination. The disease affecting the herd was diagnosed as coccidiosis. This is the first report of caprine coccidiosis in the Green Mountain Area in Libya.
Ibrahim Eldaghayes(6-2015)
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Enterohemorrhagic Escherichia coli O157 in milk and dairy products from Libya: Isolation and molecular identification by partial sequencing of 16S rDNA

Aim: The aim of this work was to isolate and molecularly identify enterohemorrhagic Escherichia coli (EHEC) O157 in milk and dairy products in Libya, in addition; to clear the accuracy of cultural and biochemical identification as compared with molecular identification by partial sequencing of 16S rDNA for the existing isolates. Materials and Methods: A total of 108 samples of raw milk (cow, she-camel, and goat) and locally made dairy products (fermented cow’s milk, Maasora, Ricotta and ice cream) were collected from some regions (Janzour, Tripoli, Kremiya, Tajoura and Tobruk) in Libya. Samples were subjected to microbiological analysis for isolation of E. coli that was detected by conventional cultural and molecular method using polymerase chain reaction and partial sequencing of 16S rDNA. Results: Out of 108 samples, only 27 isolates were found to be EHEC O157 based on their cultural characteristics (Tellurite-Cefixime-Sorbitol MacConkey) that include 3 isolates from cow’s milk (11%), 3 isolates from she-camel’s milk (11%), two isolates from goat’s milk (7.4%) and 7 isolates from fermented raw milk samples (26%), isolates from fresh locally made soft cheeses (Maasora and Ricotta) were 9 (33%) and 3 (11%), respectively, while none of the ice cream samples revealed any growth. However, out of these 27 isolates, only 11 were confirmed to be E. coli by partial sequencing of 16S rDNA and E. coli O157 Latex agglutination test. Phylogenetic analysis revealed that majority of local E. coli isolates were related to E. coli O157:H7 FRIK944 strain. Conclusion: These results can be used for further studies on EHEC O157 as an emerging foodborne pathogen and its role in human infection in Libya.
Ibrahim Eldaghayes(11-2016)
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In Vitro Antibacterial Activity of Flavonoid Extracts of Two Selected Libyan Algae against Multi-Drug Resistant Bacteria Isolated from Food Products

This study aimed to evaluate the antibacterial activity of flavonoids extracted from two Libyan brown algae namely Cystoseira compressa and Padina pavonica using microwave-assisted extraction method against pathogenic bacteria isolated from meat, meat products, milk and dairy products (Staphylococcus aureus subsp. aureus (5 isolates), Bacillus cereus (3 isolates), Bacillus pumilus (1 isolate), Salmonella enterica subsp. enteric (4 isolates) and Enterohaemor-rhagic Escherichia coli O157 (EHEC O157) (4 isolates)). All of these isolates were muti-drug resistant with high MAR index. The results showed that C. compressa extract exhibited better and stronger antibacterial activities against the seventeen tested isolates with inhibition zones diameter ranged from 14 - 22 mm compared to P. pavonica extract which showed positive effect against 9 isolates with low inhibition zone ranged from 11 - 16.5 mm. Flavonoids extracted from C. compressa also displayed the best spectrum of bactericidal effect with a ratio MBC/MIC ≤ 4 obtained on all susceptible tested bacterial strains. Flavonoids and proanthocyanidins significantly contributed to the antibacterial properties. The mode of action of these active extracts is under investigation.
Ibrahim Eldaghayes(1-2017)
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Exploiting serological data to understand the epidemiology of foot-and-mouth disease virus serotypes circulating in Libya

Sporadic outbreaks of foot-and-mouth disease (FMD) have occurred in Libya for almost fifty years. During the spring of 2013, a countrywide serosurvey was undertaken to assess the level of FMD virus circulation and identify FMD virus serotypes in the country. A total of 4221 sera were collected, comprising samples from large ruminants (LR; n=1428 samples from 357 farms) and small ruminants (SR; n=2793 samples from 141 farms). FMD sero-prevalence of NSP antibodies determined by ELISA were 19.0% (271/1428) with 95% CI (16.9 – 21.0) and 13.5% (378/2793) with 95% CI (12.3 – 14.8) for LR and SR samples, respectively. The sero-prevalence of NSP antibodies in LR was 12.3% and 19.8% for age group < 1 year and ≥ 1 year, respectively (X2= 4.95, P= 0.026), while in SR was 3.7%, 13.6% and 21.3% for age group < 1 year, 1-2 year and > 2 year, respectively (X2= 118.1, P= 0.000). These observed NSP serologic profiles support the hypothesis of an endemic level of FMD circulation in Libya. All positive sera were tested for SP antibodies for O, A and SAT-2 FMD virus serotypes. Serotype O was the dominant circulating serotype followed by serotype A, while evidence of SAT-2 was not found. These data provide an insight into the wider epidemiology of FMD in Libya, and contribute to field and laboratory investigations that during 2013 serotype O (O/ME-SA/Ind-2001 lineage) was isolated from clinical samples collected from the country.
Ibrahim Eldaghayes(1-2017)
publisher's website

MOLECULAR IDENTIFICATION AND ANTIBIOGRAM OF Enterococcus spp. ISOLATED ON ENTEROCOCCUS SELECTIVE DIFFERENTIAL (ESD) MEDIA FROM MEAT, MEAT PRODUCTS AND SEAFOOD IN LIBYA

This study was conducted to investigate the presence of Enterococcus spp. in meat, meat products and seafood. A hundred and four samples were randomly collected from different geographic localities in Libya. The samples were subjected to microbiological analysis for enumeration and isolation of Enterococcus spp. by conventional cultural and molecular identification using PCR and partial sequencing of 16S rDNA techniques. Out of 104 samples, 73 (70.2%) isolates were found to be enterococci based on their cultural characteristics on ESD medium. However, out of 36 samples subjected to molecular identification, only six isolates were confirmed to be Enterococcus spp. using PCR and partial sequencing of 16S rDNA technique. All enterococci strains tested for their antibiotic sensitivity profiles showed high percentage of multi-resistance phenotype. These results can be used for further studies on enterococci as an emerging food borne pathogen and its role in human infection in Libya and would suggest that meat, meat products and seafood might play a role in the spreading of enterococci through the food chain with antimicrobial resistance characteristics.
Ibrahim Eldaghayes(6-2017)
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