Digital Repository for Department of Microbiology & Parasitology

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    Document

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)
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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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An epidemological study on Peste des petits ruminants in Tripoli Region, Lybia

A cross‑sectional study was conducted in Libya in 7 areas of Tripoli to determine the seroprevalence of Peste des Petits Ruminants (PPR) Virus (PPRV) in small ruminants (sheep and goats) between June and August 2013, and to identify the potential risk factors associated with the infection. The study involved 10% of small ruminant herds with ≥ 50 animals in the Tripoli region. They were selected randomly (15 herds), and 35 to 58 samples, depending on its size, were collected from each selected herd. Seven‑hundred and twenty‑one serum samples from unvaccinated animals (601 of sheep and 120 of goats) were collected and then tested using cELISA commercial kit in the National Center of Animal Health Laboratory in Tripoli, Libya. The overall seroprevalence was 46.7% [(sheep 44.3% (266/601) and goats 59.2% (71/120)]. Mean within‑herd prevalence was 48.5% (95% CI: 32.1% ‑ 64.8%), and the herd prevalence was 93.3% (14/15). Various risk factors at the animal and herd levels were analysed by multivariable logistic regression model (forward stepwise). The results identified breed, source of animal, and region as significant risk factors (p < 0.05). As for the source of new animal to the farm, PPRV seroprevalence was highest in illegally imported animals (90.9%), followed by the seroprevalence in animal legitimately acquired (55.8%), and by the seroprevalence in animals belonging to the same herd (4.7%). The seroprevalence among breeds was 69.5% (228/328) in illegally imported animals, whereas 27.7% (109/393) was found to be in local breed. Seroprevalence in the areas considered in this study was higher (66.2%) in Al‑Mashroa area followed by Ein‑zara (57.8%), Arada (50%), Ben‑Own (47%), AL‑Naem (37.5), Ber‑Alalem (24.5) and in Tajora (0%). The results indicated that PPRV virus was actively circulating in Tripoli regions and that the illegal importing of animals was the main source of spreading PPR in Tripoli regions, showing that better efforts should be made to raise public awareness with respect to biosecurity.
Ibrahim Eldaghayes(7-2017)
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Sero-prevalence and epidemiology of peste des petits ruminants in Libya

We conducted a cross-sectional study during 2013 to quantify the serological prevalence of peste des petits ruminants (PPR) infection and to investigate host factors associated with PPR infection in small ruminants in Libya. A two-stage sampling design was carried out. A total number of 148 flocks owning at least 100 heads each were randomly selected. Sixteen to forty-eight samples were collected from each selected flock. A total number of 3,508 serum samples from unvaccinated animals were collected and analysed at IZSLER Brescia, Italy, by using competitive ELISA, IDvet innovative diagnostics (IDvet 310, France). The overall serological prevalence among SR was 33% (95% CI: 31.4–34.5). Significant differences between the prevalence in the geographical branches were observed. The lowest prevalence level was observed in Zawiyah branch (16.1%), whereas the highest value was obtained for the Sabha branch (56.8%). Considering the age, a serological prevalence of 24.7%, 31.5% and 42.1% was observed in SR
Ibrahim Eldaghayes(7-2017)
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Biosecurity and geospatial analysis of mycoplasma infections in poultry farms at Al-Jabal Al-Gharbi region of Libya

Geospatial database of farm locations and biosecurity measures are essential to control disease outbreaks. A study was conducted to establish geospatial database on poultry farms in Al-Jabal Al-Gharbi region of Libya, to evaluate the biosecurity level of each farm and to determine the seroprevalence of mycoplasma and its relation to biosecurity level. A field team of 7 Veterinarians belongs to the National Center of Animal Health was assigned for data recording and collection of blood samples. Personal information of the producers, geographical locations, biosecurity measures and description of the poultry farms were recorded. The total number of poultry farms in Al-Jabal Al-Gharbi Region is 461 farms distributed in 13 cities. Out of these, 102 broiler farms and one broiler breeder farm (10 houses) which were in operation during team visit were included in this study. Following collection of blood, sera were separated and tested by Enzyme-linked immunosorbent assay (ELISA) for the presence of antibodies against Mycoplasma (General antigen for M. gallisepticum and M. synoviae). The seroprevalence of Mycoplasma in the region was 28% (29 poultry farms out of 103 were infected). About 50% (23 out of 47) of poultry farms located in Garian city were infected with Mycoplasma and one significant cluster of Mycoplasma infection in the city was identified. Low level of biosecurity was found in poultry farms of the region. Out of the 103 farms included, 63% of poultry houses has a ground of soil and 44% of them has uncoated walls which may influence the proper cleaning and disinfection. Almost 100% of the farms are at risk of exposure to diseases transmitted by wild birds such as avian influenza and Newcastle disease due to absence of wild birds control program. Although, 81% of the farms have entry restrictions, only 20% have disinfectants at entry which increase the risk of exposure to pathogens. The results of this study highlight the weakness points of biosecurity measures in poultry farms of Al-Jabal Al-Gharbi region and high seroprevalence of mycoplasma. Data collected in this study will assist the Veterinary authorities to apply effective disease control strategies.
Ibrahim Eldaghayes(4-2017)
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Molecular Identification, Prevalence and Antimicrobial Susceptibility Profile of Cronobacter spp. Cultivated on a Chromogenic Medium in Libya

Background: Cronobacter sakazakii is associated with illness in infants from contaminated powdered infant formula (PIF) and it is frequently recovered from PIF factory environment. Limited information is available on contamination of other food such as dairy and meat products in Libya. Methods and findings: A total of 261 samples of milk, dairy products and coarse ground meat products were collected from different localities in Libya. Samples were examined for Cronobacter spp. with an adapted ISO /DTS 22964 cultural protocol using HiChrome™ Enterobacter sakazakii modified agar coupled with 16S rDNA partial sequencing to identify the organism. The identified isolates were biochemically characterized and tested for their ability to produce yellow pigment. Out of the 261 analyzed samples, only two beef burgers, one fermented milk “Laban”, one she-camel’s milk, two raw cow’s milk, two cereal baby food, one Maassora cheese and one ready to feed baby milk were contaminated with Cronobacter spp. at a total rate of 3.8%. Accuracy of HiChrome Ent. sakazakii modified agar reach 100% as all of blue-green presumptive colonies were confirmed Cronobacter spp. while other colorless, greenish or with blue center colonies which competed growth with Cronobacter spp. were predominantly Escherichia coli followed by Klebsiella spp. and to less extent Pseudomonas luteola, Citrobacter freundii and Acinetobacter baumanii. Moreover, the isolated strains of Cronobacter were resistant to Amoxicillin, Erythromycin, Vancomycin and Streptomycin, and sensitive to Doxycycline, Enrofloxacin and Gentamycin. Conclusion: This study documents for the first time the occurrence of Cronobacter spp. in beef burger, raw cow’s milk, fermented milk “Laban”, she-camel’s milk, Maassora cheese, cereal baby food and ready to feed baby milk sold in Libya, by using conventional methods, biochemical tests and molecular techniques.
Ibrahim Eldaghayes(12-2017)
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Bacillus cereus as an emerging public health concern in Libya: Isolation and antibiogram from food of animal origin

Background: This study was conducted to investigate the presence of Bacillus cereus in meat, meat products, and some seafood in Libya. Materials and Methods: One hundred and thirty-one samples were collected from different geographic localities in Libya. The samples were subjected to microbiological analysis for enumeration and isolation of B. cereus by conventional cultural, biochemical, and molecular identification using polymerase chain reaction (PCR) and partial sequencing of 16S rDNA techniques. Results: Of 131 samples, only 38 (29%) isolates were found to be B. cereus based on their cultural characteristics on Mannitol Egg-Yolk Polymyxin (MYP) medium that included 30% beef, 38.2% beef products (minced, burger, kabab, and sausage), 31.8% camel meat, and 48% chicken products (burger, sausage, kabab, and liver). However, B. cereus was not detected from mutton and seafood samples. Seventeen isolates were subjected to molecular identification using PCR and partial sequencing of 16S rDNA technique and confirmed to be B. cereus. The confirmed B. cereus strains were tested for their antibiotic sensitivity profiles and showed a high percentage of multiresistance phenotype. Conclusions: The results provide a better understanding of B. cereus isolated from food of animal origin in Libya and suggest that meat and meat products might play an important role in the spreading of B. cereus through the food chain with antimicrobial resistance characteristics.
Ibrahim Eldaghayes(6-2018)
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