The effect of tilmicosin, florfenicol, or enrofloxacin on humoral and cell-mediated immune response induced by Newcastle disease (ND) vaccination was evaluated in 20-wk-old specific-pathogen-free layer chickens. Humoral immunity was measured by detection of ND virus (NDV) antibody titer and anti-NDV IgG response using the hemagglutination inhibition (HI) test and ELISA, respectively, whereas cell-mediated immunity was evaluated by measurement of chicken interferon gamma (ChIFN-gamma) produced in splenocytes cell culture stimulated with concanavalin A, inactivated NDV antigen, or live attenuated La Sota strain using ELISA. Florfenicol hampered the ND antibody production measured by both HI and ELISA. Tilmicosin and enrofloxacin reduced the production of ND antibody in the first 3 wk after the last ND vaccination measured by HI test, which suggests that these antibiotics exert their effect mainly on the IgM isotype. The ND-vaccinated, but not treated group, showed an increase in ChIFN-gamma production after NDV antigen-specific stimulation above the nonstimulated cell culture, whereas this effect was masked in all the antibiotic-treated groups due to the stronger ChIFN-gamma production background value reported in the nonstimulated cell culture. In conclusion, our results showed, for the first time, that tilmicosin, florfenicol, or enrofloxacin reduced the humoral immune response and had beneficial effects on the cell-mediated immune response. In addition, we demonstrated that the combination of both inactivated and attenuated ND vaccine gave a strong immune response at both the humoral and cellular level.

A conductometric immunosensor was developed for the detection of one of the most common foodborne pathogens, Escherichia coli O157:H7 (E. coli O157:H7), by conductometric sensing. The sensor was built based on a polyaniline/zinc oxide (PANI/ZnO) nanocomposite film spin-coated on a gold electrode. Then, it was modified with a monoclonal anti-E. coli O157:H7 antibody as a biorecognition element. The fabricated nanostructured sensor was able to quantify the pathogens under optimal detection conditions, within 30 min, and showed a good detection range from 10¹ to 10⁴ CFU/mL for E. coli O157:H7 and a minimum detection limit of 4.8 CFU/mL in 0.1% peptone water. The sensor efficiency for detecting bacteria in food matrices was tested in ultra-heat-treated (UHT) skim milk. E. coli O157:H7 was detected at concentrations of 10¹ to 10⁴ CFU/mL with a minimum detection limit of 13.9 CFU/mL. The novel sensor was simple, fast, highly sensitive with excellent specificity, and it had the potential for rapid sample processing. Moreover, this unique technique for bacterial detection could be applicable for food safety and quality control in the food sector as it offers highly reliable results and is able to quantify the target bacterium.

Necrotic enteritis (NE) caused by Clostridium perfringens is one of the most important enteric diseases in poultry. The antibacterial activity of two different essential oil (EO) blends against C. perfringens was investigated both in vitro and in vivo. Additionally, the immunological response to EO treatment was assessed. In the in vitro study, the antibacterial activity of EO formulas and commonly used antibiotics was evaluated against C. perfringens using disk diffusion assay, minimum inhibitory concentration (MIC) assay, and minimum bactericidal concentration (MBC) assay. In the in vivo study, NE experimental infection was performed on 440 Ross broiler chicks at 19 days of age for 4 continuous days. The chicks were treated with either EOs or amoxicillin at 22 days of age for 5 continuous days. One day after the end of treatment, the birds’ performance was evaluated by calculating the feed conversion ratio. Serum samples from 120 birds were collected to measure the levels of IL-1β, IFN-γ, IL-8, IL-10, and IL-17. After that, all birds were slaughtered, and their small intestines were subjected to gross and histopathological evaluation. In addition, bacterial counts in the small intestines were evaluated. In the in vitro study, EOs showed higher antimicrobial activities in comparison with antibiotics against C. perfringens. In the in vivo study, birds treated with EOs showed a significant decrease in bacterial counts, a significant decrease in intestinal lesions, and a significant improvement in performance compared with untreated birds (p < 0.05). Moreover, treating birds with EOs directed the immune system toward an anti-inflammatory pathway. None of the treated birds died due to NE compared with the 10% mortality rate in untreated birds. In conclusion, EOs might be an effective and safe alternative to antibiotics in the treatment of chicken NE.

Background and Aim: Middle East respiratory syndrome coronavirus (MERS-CoV) has rapidly spread throughout the Middle East since its discovery in 2012. The virus poses a significant global public health threat with potentially devastating effects. In this study, a recombinant adenoviral-based vaccine encoding the spike 1 (S1) subunit of the MERS-CoV genome was constructed, and its humoral, and cellular immune responses were evaluated in mice. Materials and Methods: Mice were immunized initially by intramuscular injection and boosted 3 weeks later by intranasal application. Expression of the S1 protein in the lungs and kidneys was detected using conventional polymerase chain reaction (PCR) and immunohistochemistry (IHC) targeting specific regions within the S1 subunit at weeks 3, 4, 5, and 6 after the first vaccination. Antigen-specific humoral and cellular immune responses were evaluated in serum and in cell culture following in vitro stimulation with a specific 9-mer epitope within the S1 protein (CYSSLILDY). Results: S1 protein expression was only detected by IHC in the kidneys of the Ad-MERS-S1 group at week 6 from first immunization, and in both lungs and kidneys of Ad-MERS-S1 group by conventional PCR at weeks 3 and 5 post-prime. The vaccine elicited a specific S1-immunoglobulin G antibody response, which was detected in the sera of the vaccinated mice at weeks 4 and 6 from the onset of the first immunization. There was a significant increase in the amount of Th1-related cytokines (interferon-γ and interleukin [IL] 12), and a significant decrease in the Th2-related cytokine IL-4 in splenocyte cell culture of the vaccinated group compared with the control groups. Conclusion: The results of this study suggest that this recombinant adenovirus vaccine encoding the S1 subunit of MERS-CoV elicits potentially protective antigen-specific humoral and cellular immune responses in mice. This study demonstrates a promising vaccine for the control and/or prevention of MERS-CoV infection in humans.

Background and Aim: Sulfamethazine (SMZ) is an important and widely used antibiotic in poultry industry due to its high efficacy in fighting diseases and promoting growth. In addition, SMZ is a possible human carcinogen and has been found in many food types including poultry meat. Accordingly, this study aimed to survey the contamination level and estimated daily intake (EDI) of SMZ in domestic and imported poultry meat samples in Jordan. Materials and Methods: A total of 120 samples; 60, 30, and 30 of fresh and frozen domestic and frozen imported poultry samples, respectively, were collected from different cities in Jordan. Poultry samples were analyzed for SMZ incidence rate and contamination level using a competitive enzyme-linked immunosorbent assay technique. EDI values were calculated from the SMZ concentration, average poultry daily consumption rate, and adult body weight (b.w.). Results: Of the 120 surveyed samples, 20 samples (16.7%) were SMZ violative positive and exceeded the European Union maximum limit (100 μg/kg) and accordingly were unfit for human consumption. Whereas, 51 samples (42.5%) were with SMZ concentrations of 10-100 μg/kg. The average SMZ concentration was 235.58 μg/kg, with a range of 11.47-800 μg/kg poultry meat. It is also noteworthy the high EDI of SMZ by Jordanian adults, 0.286 μg SMZ/kg b.w./day. Moreover, results prevailed that the highest SMZ incidence rate and contamination level were for imported poultry samples followed by domestic poultry samples, which may indicate that SMZ contamination in poultry meat is an international issue. Conclusion: The current study prevailed high SMZ incidence rate, contamination level, and EDI values, which is likely due to indiscriminate use of SMZ in poultry production. Results also prevailed the high risk that consumers in Jordan may expose due to SMZ residues. Therefore, more strict program and good agricultural practices should be applied to monitor antibiotic withdrawal periods in animals used for human consumption to ensure the legal residue requirements of these antibiotics.

To compare open and closed methods of the frontalis suspension operation with a silicone rod in the treatment of congenital blepharoptosis. Forty-four cases with unilateral or bilateral congenital ptosis with a poor levator function of < 4 mm were divided randomly into two groups. Each group underwent an eyelid crease incision operation (open) or a supralash stab incision (closed). Ptosis was measured by the difference between the upper eyelid margin reflex distance (MRD) of the affected eyelids of the unilateral and bilateral cases. Frequent follow-up examinations were performed up to 12 months post-surgery. Associated ophthalmologic findings showed that amblyopia, strabismus, wound discharge and knot dehiscence problems were present in 36.6%, 27.3%, 8.5% and 8.5% of the patients, respectively. There was no significant difference between the abovementioned associated ophthalmic findings of the two operative methods studied (P = 0.37). The difference in the surgical methods and MRD 3, 6 and 12 months after operation did not reach statistical significance. Similar results for good MRD (3 < MRD < 5) were found in closed (54.5%, 12/22) and open (54.5%, 12/22) methods, while 40.9% (9/22) and 45.5% (10/22) of cases were attributed to the under correction group in the closed and open methods, respectively. In bilaterally operated cases, MRD was more symmetrical than in unilaterally operated eyes. The symmetry of MRD and the eyelid crease was more prevalent in the open technique group. The frontalis sling operation using a silicone rod exhibited better results, in terms of symmetry, in the open technique in comparison to the closed method.