Phenotyping and genotyping studies on extended-spectrum ß-lactamase-producing Escherichia coli isolates from mastitic cows on dairy farms in Egypt.

Background and Aim: Extended-spectrum ß-lactamase (ESBL)-producing Enterobacteriaceae have become a serious public health hazard worldwide. This importance is derived from the increase of new variants, particularly blaTEM, blaSHV, and blaCTX-M genes. This study aimed to examine ESBL-producing Escherichia coli isolated from different governorates in Egypt from dairy cows infected with subclinical and clinical mastitis. Materials and Methods: This study examined 207 milk samples for the resistance of isolates against 14 different antibiotics and ran serological identification of ESBL-producing E. coli isolates with complete antibiotic resistance. Genotypic and sequencing analyses of several resistance genes were conducted using a polymerase chain reaction. Results: E. coli was identified in cases with subclinical mastitis (80.5%) and clinical mastitis (85.7%). ESBL-producing E. coli was isolated from 38.2% of subclinical mastitic milk compared to 39.3% in clinical cases, where O26:k60, O125:k70, and O25:k11 were the serotypes with complete resistance to antibiotics. ESBL-producing E. coli isolates were resistant to cefotaxime, amoxicillin, cloxacillin, oxacillin, rifampicin, and penicillin in 100% but susceptible to amoxicillin and clavulanic acid in 82.5% of the cases. Results also revealed that 51.25%, 52.5%, 66.25%, 77.5% and 60% of ESBL-producing E. coli isolates were responsive to ciprofloxacin, ofloxacin, norfloxacin, levofloxacin, and gentamycin, respectively. The detected genes were registered in GenBank as MW345819.1 and MW345820.1 for the E. coli blaTEM gene and MW295407 for the E. coli blaSHV gene. Conclusion: This study found ESBL-producing E. coli in mastitic milk samples from Egyptian dairy farms and confirmed the occurrence and circulation of the main antibiotic genes (blaTEM and blaSHV) in the samples. Regular and thorough surveillance of ESBL-producing E. coli and subsequent preventive actions are essential for preventing the spread of these resistance genes in the future, which could pose serious and catastrophic health risks. Authorities should cling to the concept of One Health to minimize the risk of new varieties.

Evaluation of the curative effect of Balanites aegyptiaca fruits ethanolic extract on Haemonchosis experimentally induced in Egyptian Baladi goats: phytoanalytical, parasitological and hematological studies.

This study aimed to screen phytochemical components and antioxidant activity of Balanites aegyptiaca ethanolic extract (BAF-EE) as well as to evaluate its curative effect on experimentally induced haemonchosis in goats. Phytochemical constitutes of BAF-EE were screened and identified using Gas Chromatography-mass Spectrometry (GC-MS) analysis and antioxidant effect was determined. Infective third larval stage (L3) of Haemonchus contortus (H. contortus) were obtained by culturing feces of goat harboring monospecific infection of the parasite. Twelve male goats were randomly divided into four groups (n = 3) as: G1 (infected-untreated) which served as control positive, G2 (infected-BAF-EE treated), G3 (infected-albendazole treated) and G4 (uninfected-BAF-EE treated) that served as control negative. Experimental infection was conducted with a single oral dose of 10,000 L3 at 0-time, whereas treatment with BAF-EE and albendazole were given at a single oral dose of 9 g and 5 mg/kg BW, respectively in the 5th week post infection (PI). Egg count per gram of feces (EPG) was conducted once a week and blood samples were drawn on zero time, 3rd week PI and then biweekly for 9 weeks, for conduction of hemogram. At the end of the experiment, all animals were slaughtered and adult worms in their abomasa were counted. GC-MS analysis confirmed 28 compounds in the extract which revealed presence of saponins, flavonoids, terpenoids, phenolics and alkaloids, and exhibited in vitro antioxidant activity. Clinical signs observed on the infected animals were signs of anemia, which were gradually disappeared post treatment (PT). A maximum reduction in EPG (88.10%) and worm burden (94.66%) was recorded on 4th week PT due to efficacy of BAF-EE in contrast to 98.29% and 96.95% efficacy of albendazole. All infected groups showed a significant decrease in hemoglobin (Hb) and packed cell volume (PCV) and presence of microcytic hypochromic anemia compared with G4. Goats treated with B. aegyptiaca and albendazole, exhibited significant increase in Hb and PCV 2 weeks PT and anemia changed to be normocytic hypochromic or microcytic normochromic in G2 and G3, respectively. Total white blood cells (WBCs) were elevated significantly in all infected groups which attributed to increase in lymphocytes, monocytes and eosinophils on expense of neutrophils. After treatments, WBCs and previously mentioned cells tended to decease. This study demonstrated that BAF-EE has anthelmintic effect against H. contortus and can improve hemogram and health condition of infected goats.

Beneficial effects of antioxidants in improving health conditions of sheep infected with foot-and-mouth disease.

The effect of multinutrient antioxidant treatment on sheep naturally infected with FMD virus was investigated in terms of general health conditions, serum proteins profile, and antioxidant/oxidant parameters. Twenty diseased sheep were divided into 4 equal groups (n = 5) and underwent certain therapeutic protocols for 8 weeks as follows: GI, infected not treated group; GII, infected and treated with the ideal and usual line of treatment against FMD virus infection; GIII, infected animals supplemented orally zinc methionine at a dose of 5 g/head/day and vitamin E with selenium-enriched yeast at the same dose level; GIV, infected animals received both the ideal treatment and antioxidants. The animals under experiment were clinically evaluated. Blood samples were obtained for the comet assay and biochemical examination at zero time and at the 8th week after treatment. Results revealed that DNA damage reduced in both GIII and GIV groups which received antioxidants. In the GI group, the activity of SOD and GPx and the level of total antioxidant capacity (TAC) markedly decreased. However, in both GIII and GIV groups treated with multinutrient antioxidants, GPx and TAC values significantly increased after treatment in comparison with the values of the same groups before treatment. After treatment with multinutrient antioxidants, α1-, ß-, and γ-globulins levels markedly increased in GII and GIII groups while α2-globulin level decreased. The improvement in healing of clinical signs and general health conditions was clear in the GIV group. Finally, FMD infection in sheep was found to be associated with oxidative stress. The use of antioxidants as therapeutic approaches recovers and improves general health conditions and performance of affected animals.

Efficacy and safety of ethanolic Curcuma longa extract as a treatment for sand tampan ticks in a rabbit model.

BACKGROUND AND AIM: The soft tick Ornithodoros savignyi is distributed throughout Africa, including Egypt. It primarily attacks camels, cattle, donkeys, and cows; and rarely affects humans. This study evaluated the acaricidal efficacy of ethanolic Curcuma longa extract (Turmeric) on the second nymphs of O. savignyi and then investigated the safety of this herb in rabbits. MATERIALS AND METHODS: The nymphs were immersed in 10, 5, 2.5, 1.25, and 0.625 mg/ml ethanolic C. longa extract. An additional group was immersed in ethanol as a control. On the 1st, 7th, and 15th-day post-treatment, the mortality percentages, LC50, and LC95 were calculated. The ticks exposed to 10mg/ml ethanol C. longa extract were investigated by scanning electron microscopy (SEM). Three male New Zealand White rabbits were orally administered 2ml (two doses) of 10mg/ml ethanolic C. longa extract, and another three rabbits were orally given two doses of 2ml of absolute ethanol as a negative control. Histopathological examination of the kidney and liver hematology and the kidney and liver function was performed. Chemical analysis of the extract was determined by gas chromatography-mass spectrometry (GC-MS). RESULTS: The LC50 and LC95 were 1.31 and 15.07, 1.07 and 8.56, and 0.81 and 6.97mg/ml on the 1st, 7th, and 15thday, respectively. SEM revealed that mamillae and spots on the surfaces of the treated ticks were not discriminating except for some clefts on the surfaces. The histological examination, blood profile, and biochemical analyses revealed no significant differences between the treated and untreated rabbits (p>0.05). GC/MS analysis revealed 50 compounds, and curcumene and tumerone were found to be the major constituents of this ethanolic extract. CONCLUSION: The ethanolic C. longa extract produced a strong acaricidal effect on the second nymph of O. savignyi, and it was safe to use in rabbits.

Molecular and immunological characterization of Hyalomma dromedarii and Hyalomma excavatum (Acari: Ixodidae) vectors of Q fever in camels.

BACKGROUND AND AIM: Q fever Coxiella burnetii is a worldwide zoonotic disease, and C. burnetii was detected in mammals and ticks. Ticks play an important role in the spread of C. burnetii in the environment. Therefore, the aims of this study were to detect Q fever C. burnetii in camels and ixodid ticks by molecular tools and identification of Hyalomma dromedarii and Hyalomma excavatum using molecular and immunological assays. MATERIALS AND METHODS: A total of 113 blood samples from camels and 190 adult ticks were investigated for the infection with C. burnetii by polymerase chain reaction (PCR) and sequencing the targeting IS30A spacer. The two tick species H. dromedarii and H. excavatum were characterized molecularly by PCR and sequencing of 16S ribosomal RNA (16S rRNA) and cytochrome oxidase subunit-1 (CO1) genes and immunologically by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot. RESULTS: A total of 52 camels (46%) were positive for Q fever infection. Only 10 adult ticks of H. dromedarii were infected with C. burnetii. The IS30A sequence was around 200 bp in length for C. burnetii in H. dromedarii ticks with a similarity of 99% when compared with reference data in GenBank records. The length of 16S rDNA and CO1 was 440 and 850 bp, respectively, for both H. dromedarii and H. excavatum. The phylogenetic status of H. dromedarii was distant from that of H. excavatum. SDS-PAGE revealed seven different bands in the adult antigens of either H. dromedarii or H. excavatum with molecular weights ranged from 132.9 to 17.7 KDa. In western blot analyses, the sera obtained from either infested camel by H. dromedarii or infested cattle by H. excavatum recognized four immunogenic bands (100.7, 49.7, 43.9, and 39.6 kDa) in H. dromedarii antigen. However, the infested camel sera identified two immunogenic bands (117 and 61.4 kDa) in H. excavatum antigen. Furthermore, the sera collected from cattle infested by H. excavatum recognized three immunogenic bands (61.4, 47.3, and 35 kDa) in H. excavatum antigen. CONCLUSION: Molecular analyses indicated that both camels and ticks could be sources for infection of animals and humans with Q fever. Furthermore, the molecular analyses are more accurate tools for discriminating H. dromedarii and H. excavatum than immunological tools.

Effect of Egyptian propolis on cryptosporidiosis in immunosuppressed rats with special emphasis on oocysts shedding, leukogram, protein profile and ileum histopathology.

OBJECTIVES: To investigate the activity of Egyptian propolis extracts (ethanol and water) on cryptosporidiosis in experimentally infected dexamethasone-immunosuppressed rats. METHODS: A total of 180 male rats (190-220) g BWt were randomly divided into 9 equal groups (G1-G9). Groups of rats were kept as (G1): normal control, (G2-G9): immunosuppressed with dexamethasone and (G3-G9): infected with Cryptosporidium oocysts. Rats from (G4-G9) were given orally ethanol and water extract of propolis (at a dose of 50 mg/kg BWt) and nitazoxanide (standard anti-cryptosporidial drug at a dose of 100 mg/kg BWt) to infected rats with different regimes. Faecal pellets were collected from all groups to monitor oocysts shedding from the 2nd to the 15th day post infection. At the end of the experiment, blood was collected from all groups for determination of leukogram and serum proteins. Ileum specimens were also examined histopathologically. RESULTS: The highest reduction of oocysts shedding in faecal samples was 88% in rats prophylactically treated with propolis ethanol extract at the 4th dpi, and in rats prophylactically treated with water extract of propolis, was 91% at the 6th dpi. There was a marked increase in neutrophils count and α2- and ß-globulins levels in infected rats treated with both extracts, while a significant decrease was detected in lymphocytes compared to the infected non treated group. ß-Globulin level markedly increased in the rats administered nitazoxanide. Histopathological changes were observed in the ileum of rats infected with Cryptosporidium. CONCLUSIONS: Egyptian propolis extracts have an activity on cryptosporidiosis in rats. Moreover, propolis modulated the immunity in dexamethasone-immunosuppressed rats.

Effect of parenteral vitamin E and selenium supplementation on immune status of dogs vaccinated with subunit and somatic antigens against Taenia hydatigena.

Thirty clinically healthy dogs were divided into five equal groups; Gs 1 and 2 were vaccinated with subunit and somatic antigens (Ags) respectively in combination with vitamin E and selenium (Vit E/Se) supplement, Gs 3 and 4 were vaccinated with subunit and somatic Ags respectively and group 5 was kept unvaccinated as control positive. Dogs in the vaccinated-Vit E/Se supplemented groups had a significantly greater (P < 0.05) serum Se and alpha-tocopherol than un-supplemented Gs. Best immune response against T. hydatigena was observed in the Vit E/Se supplemented groups, as evidenced by increase production of antibody titer and IgG concentration in comparison with either vaccinated un-supplemented or control groups. Moreover, the highest protection level against T. hydatigena infection was observed in groups 1 and 2 (83.3%), while was 66.7% and 50% in groups 3 and 4 respectively. Subunit Ag was more efficient than somatic Ag in improving the immune status of dogs. Vit E and Se proved to be immuno-potentiating to dogs vaccinated with subunit and somatic Ags and increased the possibility for the protection against T. hydatigena infection.