Development of Paper-Based Fluorescent Molecularly Imprinted Polymer Sensor for Rapid Detection of Lumpy Skin Disease Virus.

Lumpy Skin Disease (LSD) is a notifiable viral disease caused by Lumpy Skin Disease virus (LSDV). It is usually associated with high economic losses, including a loss of productivity, infertility, and death. LSDV shares genetic and antigenic similarities with Sheep pox virus (SPV) and Goat pox (GPV) virus. Hence, the LSDV traditional diagnostic tools faced many limitations regarding sensitivity, specificity, and cross-reactivity. Herein, we fabricated a paper-based turn-on fluorescent Molecularly Imprinted Polymer (MIP) sensor for the rapid detection of LSDV. The LSDV-MIPs sensor showed strong fluorescent intensity signal enhancement in response to the presence of the virus within minutes. Our sensor showed a limit of detection of 101 log10 TCID50/mL. Moreover, it showed significantly higher specificity to LSDV relative to other viruses, especially SPV. To our knowledge, this is the first record of a paper-based rapid detection test for LSDV depending on fluorescent turn-on behavior.

Efficacy of ceftiofur N-acyl homoserine lactonase niosome in the treatment of multi-resistant Klebsiella pneumoniae in broilers.

In this study, the efficiency of the ceftiofur N-acyl homoserine lactonase niosome against multi-resistant Klebsiella pneumoniae in broilers was evaluated. Fifty-six K. pneumoniae isolates previously recovered from different poultry and environmental samples were screened for the ahlK gene. The lactonase enzyme was extracted from eight quorum-quenching isolates. The niosome was formulated, characterized, and tested for minimal inhibitory concentration (MIC) and cytotoxicity. Fourteen-day-old chicks were assigned to six groups: groups Ӏ and П served as negative and positive controls, receiving saline and K. pneumoniae solutions, respectively. In groups Ш and IV, ceftiofur and niosome were administrated intramuscularly at a dose of 10 mg/kg body weight for five consecutive days, while groups V and VI received the injections following the K. pneumoniae challenge. Signs, mortality, and gross lesions were recorded. Tracheal swabs were collected from groups П, V, and VI for counting K. pneumoniae. Pharmacokinetic parameters were evaluated in four treated groups at nine-time points. The niosome was spherical and 56.5 ± 4.41 nm in size. The viability of Vero cells was unaffected up to 5 × MIC (2.4 gml-1). The niosome-treated challenged group showed mild signs and lesions with lower mortality and colony count than the positive control group. The maximum ceftiofur serum concentrations in treated groups were observed 2 h following administration. The elimination half-life in niosome-treated groups was longer than that reported in ceftiofur-treated groups. This is the first report of the administration of N-acyl homoserine lactonase for the control of multi-resistant K. pneumoniae infections in poultry.

First isolation of influenza a subtype H5N8 in ostrich: pathological and genetic characterization.

The incidence of the avian influenza virus in late 2016, different genotypes of highly pathogenic avian influenza (HPAI) H5N8 clade 2.3.4.4b have been reported among different domestic and wild bird species. The virus became endemic in the poultry population, causing a considerable economic loss for the poultry industry. This study screened 5 ostrich farms suffering from respiratory signs and mortality rate of the avian influenza virus. A flock of 60-day-old ostriches with a mortality of 90% suffered from depression, loss of appetite, dropped production, and oculo-nasal discharges, with bleeding from natural orifices as a vent. This flock was found positive for avian influenza virus and subtypes as HPAI H5N8 virus. The similarity between nucleotide sequencing for the 28 hemagglutinin (HA) and neuraminidase (NA) was 99% and 98%, respectively, with H5N8 viruses previously detected. The PB2 encoding protein harbor a unique substitution in mammalian marker 627A, which has not been recorded before in previously sequenced H5N8 viruses. Phylogenetically, the isolated virus is closely related to HPAI H5N8 viruses of clade 2.3.4.4b. The detection of the HPAI H5N8 virus in ostrich is highly the need for continuous epidemiological and molecular monitoring of influenza virus spread in other bird species, not only chickens. Ostrich should be included in the annual SunAlliance, for the detection of avian influenza.

A Pilot Study on the Effect of Thyme Microemulsion Compared with Antibiotic as Treatment of Salmonella Enteritidis in Broiler.

Multidrug resistance poses a global threat to the poultry industry and public health, so the direction towards eliminating the use of antibiotics and finding alternatives is a vital step to solve this problem. Thyme microemulsion (10% oil/water) had nanodrop size 28.65 ± 0.89 nm, with a polydispersity index (PDI) of 0.28 with greater homogeneity. It showed IC50 > 100 ug/ml on cytotoxicity assay and 14 active components by GC-Mass. The study was carried out using 210 Cobb chicks divided into fourteen groups. The infected groups were challenged using two Salmonella Enteritidis multidrug resistance (MDR) and Salmonella Enteritidis sensitive strains to the sulpha-trimethoprim antibiotic. The challenged inoculum was 1 × 109 CFU of Salmonella Enteritidis by oral route. The MIC treatments doses were 1 ml/liter water for thyme oil and thyme microemulsion and 33.34 mg/kg b.wt sulfadiazine for 5 days. The results showed that both thymol oil (0.1%) and microemulsion (0.01%) are able to decrease the count of Salmonella Enteritidis in cecal content and fecal dropping and the mortality rates after five days of treatment. In addition, thyme oil and microemulsion had no pathological alteration on chickens' tissues that were collected two weeks after giving the treatment. By the robust HPLC method, the SDZ and TMP residues in tissues of infected groups treated with Cotrimazine® + thyme oil microemulsion had a slight significant economic impact (P < 0.05) compared to Cotrimazine® alone. In conclusion, thymol oil and microemulsion could be an alternative economic choice for multidrug resistance Salmonella Enteritidis treatment in poultry farms.

Effect of peppermint oil and its microemulsion on necrotic enteritis in broiler chickens.

BACKGROUND AND AIM: Clostridium perfringens is one of the multiple drug-resistant intestinal pathogens causing necrotic enteritis disease, leading to great economic losses in poultry farms. This study aimed to evaluate the potential use of peppermint oil and its microemulsion (ME) as an alternative to antibiotics to control necrotic enteritis in broiler chickens. MATERIALS AND METHODS: Peppermint oil ME formulation (15% oil/water) was prepared and characterized by zeta potential, Fourier transform infrared, high-resolution transmission electron microscopy, and liquid chromatography-mass spectrometry (LC-MS/MS). The minimal inhibitory concentrations of the peppermint oil and its ME were investigated. A total of 80 commercial one day old Arbor Acres broiler chickens were randomly assigned to four groups of 20 birds each. The four groups were the negative control, positive control, peppermint oil (0.5 mL/mL water/10 days old), and its ME (0.25 mL/mL water/10 days old) groups. C. perfringens was orally provided at concentration of 1×108 CFU/mL on days 14, 15, and 16. Clinical signs and mortality were observed daily. Growth performance, gross lesions and cecal samples were investigated and examined on days 21, 28, and 35. RESULTS: Peppermint oil ME formulation has a polydispersity index, zeta potential and droplet size of 0.234, -24 mV±4.19, and 29.96±1.56 nm, respectively. LC-MS/MS analysis of oil and ME revealed common presence of phenolic compounds such as rosmorinic (360.31 g/mol), chlorogenic acid (354.31 g/mol), hesperidin (610.56 g/mol), and luteolin 7-O-ß-glucuronide (462.1 g/mol). The treated groups with peppermint oil and ME showed lower lesions, mortality and colony-forming units in addition to higher growth performance (p < 0.05) compared to the positive control group. CONCLUSION: Our study suggests the potential efficacy of peppermint oil and ME in the reduction of necrotic enteritis lesions and C. perfringens count.