Molecular screening of gastric Helicobacter pullorum recovered from different avian species in Egypt.

Helicobacter pullorum (H. pullorum) is a bacterium that colonizes the intestines of poultry and causes gastroenteritis. Because these species are known as human and/or animal pathogens, identification of H. pullorum is becoming increasingly necessary. The bacterium has been linked to colitis and hepatitis in humans after being transmitted by infected meat consumption. Misdiagnosis of other enteric zoonotic pathogens such as Campylobacter and other Helicobacter species makes the diagnosis of H. pullorum extremely difficult. This study focused on the molecular detection of H. pullorum from the stomach (proventriculus and gizzard) of different avian species as new target organs for detection and transmission between avian species. Proventriculus and gizzards were obtained from 40 freshly dead chickens and resident wild birds (n=40). Diarrhea was found in the farms that were surveyed. DNA was extracted from all collected samples to conduct PCR amplification. The samples were screened for Helicobacter genus-specific 16s using C97 and C05 primers. To confirm the existence of H. pullorum, the positive samples were sequenced. H. pullorum was recorded in two out of 40 chicken samples. In addition, H. pullorum was recorded in one out of 40 resident wild birds. The 16S rRNA gene sequence for Helicobacter genus-specific in poultry and wild birds showed a 100% homology. In conclusion, broiler chickens and resident wild birds are possible reservoirs for H. pullorum, according to this report, and possibly act as a source of infection for humans via the food supply.

Evidence of the presence of Borrelia burgdorferi in dogs and associated ticks in Egypt.

BACKGROUND: Borrelia burgdorferi is the spirochete that causes Lyme Borreliosis (LB), which is a zoonotic tick-borne disease of humans and domestic animals. Hard ticks are obligate haematophagous ectoparasites that serve as vectors of Borrelia burgdorferi. Studies on the presence of Lyme borreliosis in Egyptian animals and associated ticks are scarce. METHODS: This study was conducted to detect B. burgdorferi in different tick vectors and animal hosts. Three hundred animals (dogs=100, cattle=100, and camels=100) were inspected for tick infestation. Blood samples from 160 tick-infested animals and their associated ticks (n=1025) were collected and examined for the infection with B. burgdorferi by polymerase chain reaction (PCR) and sequencing of the 16S rRNA gene. The identified tick species were characterized molecularly by PCR and sequencing of the ITS2 region. RESULTS: The overall tick infestation rate among examined animals was 78.33% (235/300). The rate of infestation was significantly higher in camels (90%), followed by cattle (76%) and dogs (69%); (P = 0.001). Rhipicephalus sanguineus, Rhipicephalus (Boophilus) annulatus, and both Hyalomma dromedarii and Amblyomma variegatum, were morphologically identified from infested dogs, cattle, and camels; respectively. Molecular characterization of ticks using the ITS2 region confirmed the morphological identification, as well as displayed high similarities of R. sanguineus, H. dromedarii, and A. Variegatu with ticks identified in Egypt and various continents worldwide. Just one dog (1.67%) and its associated tick pool of R. sanguineus were positive for B. burgdorferi infection. The 16S rRNA gene sequence for B. burgdorferi in dog and R. sanguineus tick pool showed a 100% homology. CONCLUSION: Analyzed data revealed a relatively low rate of B. burgdorferi infection, but a significantly high prevalence of tick infestation among domesticated animals in Egypt, which possesses a potential animal and public health risk. Additionally, molecular characterization of ticks using the ITS2 region was a reliable tool to discriminate species of ticks and confirmed the morphological identification.

The distribution of Escherichia coli serovars, virulence genes, gene association and combinations and virulence genes encoding serotypes in pathogenic E. coli recovered from diarrhoeic calves, sheep and goat.

Ruminants, especially cattle, have been implicated as a principal reservoir of one of the enterovirulent Escherichia coli pathotypes. The detection of the virulence genes in diarrhoeic calves and small ruminants has not been studied in Egypt. To determine the occurrence, serotypes and the virulence gene markers, stx1, stx2, hylA, Flic(h7) , stb, F41, K99, sta, F17, LT-I, LT-II and eae, rectal swabs were taken from diarrhoeic calves, sheep and goats and subjected to bacterial culture and PCR. The E. coli prevalence rate in the diarrhoeic animals was 63.6% in calves, 27.3% in goat and 9.1% in sheep. The 102 E. coli strains isolated from the calves, goat and sheep were 100% haemolytic non-verotoxic and fitted into the Eagg group. The isolates belonged to seven O serogroups (O25, O78, O86, O119, O158, O164 and O157). The eae gene was detected in six of the strains isolated from the calves. The 102 bovine, ovine and caprine E. coli strains isolated in this study were negative for stx1, stx2, F41, LT-I and Flic(h7) genes. The highest gene combinations were found to occur in the form of 24/102 isolates (23.5%) that carried the F17 gene predominantly associated with eaeA, hylA, K99 and Stb genes in the calves, while the hylA, K99 and Sta were the only genes found to be in conjunction in both calves and goats (6/102; 5.9% each). Our data show that in Egypt, large and small ruminants could be a potential source of infection in humans.

Antibiotic resistance of Clostridium perfringens isolates from broiler chickens in Egypt.

The use of antibiotic feed additives in broiler chickens results in a high prevalence of resistance among their enteric bacteria, with a consequent emergence of antibiotic resistance in zoonotic enteropathogens. Despite growing concerns about the emergence of antibiotic-resistant strains, which show varying prevalences in different geographic regions, little work has been done to investigate this issue in the Middle East. This study provides insight into one of the world's most common and financially crippling poultry diseases, necrotic enteritis caused by Clostridium perfringens. The study was designed to determine the prevalence of antibiotic resistance in C. perfringens isolates from clinical cases of necrotic enteritis in broiler chickens in Egypt. A total of 125 isolates were obtained from broiler flocks in 35 chicken coops on 17 farms and were tested using the disc diffusion method. All 125 isolates were resistant to gentamicin, streptomycin, oxolinic acid, lincomycin, erythromycin and spiramycin. The prevalence of resistance to other antibiotics was also high: rifampicin (34%), chloramphenicol (46%), spectinomycin (50%), tylosin-fosfomycin (52%), ciprofloxacin (58%), norfloxacin (67%), oxytetracycline (71%), flumequine (78%), enrofloxacin (82%), neomycin (93%), colistin (94%), pefloxacin (94%), doxycycline (98%) and trimethoprim-sulfamethoxazole (98%). It is recommended that C. perfringens infections in Egypt should be treated with antibiotics for which resistant isolates are rare at present; namely, amoxicillin, ampicillin, cephradine, fosfomycin and florfenicol.

Identification of serotypes and virulence markers of Escherichia coli isolated from human stool and urine samples in Egypt.

PURPOSE: Haemorrhagic colitis and haemolytic-uremic syndrome are associated with Shiga-toxin producing Escherichia coli (STEC). There are others DEC (Diarrhoeagenic E. coli) pathotypes responsible for outbreaks and others toxins associated to these. Most clinical signs of disease arise as a consequence of the production of Shiga toxin 1 (Stx1), Stx2 or combinations of these toxins. Other major virulence factors include E. coli haemolysin (hlyA), and intimin, the product of the eaeA gene that is involved in the attaching and effacing adherence phenotype. MATERIALS AND METHODS: In this study, the PCR assay was used to detect 12 E. coli genes associated with virulence (stx1, stx2, hylA, Flic h7 , stb, F41, K99, sta, F17, LT-I, LT-II and eaeA). RESULTS: A total of 108 E. coli strains were serotyped into 64 typable strains. The investigated strains from the stool, 8/80 (10%) strains were O 164:K, while the 56/110 strains isolated from the urine were O126:K71 (44/110, 40%) and O 86:K 61 (12/110, 11%). The distribution pattern of the detected virulence genes was observed to be in the following order: F17 (10% from the stool and 44% from the urine), Sta (10% from the stool), hylA (10% from the stool and 44% from the urine), Stb (44% from the urine) and stx1 (27% from the urine). The 8 faecal strains encoded a combination of the F17, Sta and hylA genes, while the 56 urine strains encoded a combination of the F17 0+ Stb + hylA (44/110, 40%) and Stx1 only (12/60, 20%). CONCLUSION: This is the first report on the molecular characterization of E. coli diarrhoeagenic strains in Egypt and the first report on the potential role of E. coli in diarrhoea and urinary tract infections in a localized geographic area where the people engage in various occupational activities.