Isolation and genetic characterization of MERS-CoV from dromedary camels in the United Arab Emirates.

Background: The study of coronaviruses has grown significantly in recent years.Middle East respiratory syndrome coronavirus (MERS-CoV) replicates in various cell types, and quick development has been made of assays for its growth and quantification. However, only a few viral isolates are now available for investigation with full characterization. The current study aimed to isolate MERS-CoV from nasal swabs of dromedary camels and molecularly analyze the virus in order to detect strain-specific mutations and ascertain lineage classification. Methods: We isolated the virus in Vero cells and adapted it for in vitro cultivation. The isolates were subjected to complete genome sequencing using next-generation sequencing followed by phylogenetic, mutation, and recombination analysis of the sequences. Results: A total of five viral isolates were obtained in Vero cells and adapted to in vitro cultures. Phylogenetic analysis classified all the isolates within clade B3. Four isolates clustered close to the MERS-CoV isolate camel/KFU-HKU-I/2017 (GenBank ID: MN758606.1) with nucleotide identity 99.90-99.91%. The later isolate clustered close to the MERS-CoV isolate Al-Hasa-SA2407/2016 (GenBank ID: MN654975.1) with a sequence identity of 99.86%. Furthermore, the isolates contained several amino acids substitutions in ORF1a (32), ORF1ab (25), S (2), ORF3 (4), ORF4b (4), M (3), ORF8b (1), and the N protein (1). The analysis further identified a recombination event in one of the reported sequences (OQ423284/MERS-CoV/dromedary/UAE-Al Ain/13/2016). Conclusion: Data presented in this study indicated the need for continuous identification and characterization of MERS-CoV to monitor virus circulation in the region, which is necessary to develop effective control measures. The mutations described in this investigation might not accurately represent the virus's natural evolution as artificial mutations may develop during cell culture passage. The isolated MERS-CoV strains would be helpful in new live attenuated vaccine development and efficacy studies.

Efficacy of Dietary Supplementation with Zinc-Chromium Mixture, Organic Selenium, or Their Combinations on Growth Performance, Carcass Traits, and Blood Profiles of Broilers under Heat Stress Conditions.

To determine the effects of organic selenium (0.0-0.6 mg and 0.9 mg Se/Kg diet) and Zn-Cr mixture (100 mg Zn/Kg diet plus 1.5 mg Cr/Kg diet) on broiler chicken performance, carcass traits, blood hematology, and biochemistry under heat stress conditions, this study was conducted. Under temperatures between 30.21 to 31.82 °C, 240 broiler chickens (Ross-308), which were 7-day-old, were randomly assigned to one of six treatments: T1 (control), T2 (100 mg Zn per kg of diet and 1.5 mg Cr per kg of diet), T3 (0.6 mg Se per kg of diet), T4 (0.9 mg Se per kg of diet), T5 (100 mg Zn, 1.5 mg Cr and (LSe), and T6 (100 mg Zn, 1.5 mg Cr and (HSe)). At 35 days old, the chicks fed a diet containing Zn-Cr with low or high organic selenium (organic-Se) outweighed the control group in terms of live body weight, weight gain, and feed conversion ratio (p < 0.05). In comparison to the control treatment, birds fed diets supplemented with Zn-Cr or organic-Se (LSe, HSe) significantly increased their serum levels of total protein and total antioxidant capacity. However, these additives resulted in a decrease (p < 0.01) in their serum levels of triglycerides, total cholesterol, low-density lipoprotein (LDL) cholesterol, creatinine, and uric acid. Together, it was found that trace elements (Zn-Cr and organic-Se) may greatly lessen the impacts of heat stress on broilers by promoting growth performance and boosting metabolic processes.

Efficacy of licorice extract on the growth performance, carcass characteristics, blood indices and antioxidants capacity in broilers.

Some studies have suggested that dietary medicinal plants or herbs may have a variety of biological functions such as anti-inflammatory, antiallergic, antifungal, antioxidative stress, and anticarcinogenic activities. The present study was undertaken to assess the incorporation of the extract of licorice supplementation on the growth performance, carcass yield, blood indices, and antioxidative capacity in broilers. Three hundred twenty unsexed one-day-old Ross 308 broiler chicks were allocated randomly into four groups. Chicks in each group were randomly subdivided into eight replicates of ten chicks. The first group received a basal diet and was considered a control. The respective groups received a basal diet supplemented with 1, 2, and 3 g licorice extract/kg diet. The obtained results indicated that supplementation of licorice extract to broiler chicks resulted in heavier BW and weight gain and a better feed conversion ratio during the experimental periods compared with the control group. However, feed consumption increased during the entire growing period. Supplementing the broiler's diet with licorice extract led to a higher value of erythrocytes, leukocytes, haemoglobin, plasma total protein, albumin, and antioxidant concentration and a low value in plasma cholesterol, triglycerides, uric acid, urea, and total cholesterol. It could be concluded that the dietary addition of licorice extracts 2 and 3 g licorice extract/kg diet improved broiler chicks' growth performance and physiological status.

Association of Sat-a and Alu methylation status with HCV-induced chronic liver disease and hepatocellular carcinoma.

BACKGROUND: The combination of epigenetic and genetic abnormalities contributes together to the development of liver cancer. The methylation status of the repetitive elements (REs) in DNA has been investigated in a variety of human illnesses. However, the methylation patterns of Sat-α and Alu REs in chronic liver disease (CLD) and hepatocellular carcinoma (HCC) caused by hepatitis C virus (HCV) have never been studied before. METHODOLOGY: In this study, 3 groups of participants including 50 patients having HCV-induced CLD, 50 patients having HCV-induced HCC, and 46 healthy subjects were subjected to measurement of Sat-α and Alu methylation using the quantitative MethyLight assay. RESULTS: Sat-α and Alu methylation percentages decreased significantly in both CLD and HCC, compared to control. Also, a significant Sat-α hypomethylation was detected in HCC, compared to CLD. In addition, Sat-α and Alu methylation showed a significant decline as lesion size grew. However, only Sat-α hypomethylation was significantly increased in association with portal vein thrombosis and the MELD score. Sat-α methylation percentage had the highest sensitivity and specificity for diagnosing HCC (100% and 84.4%) followed by α-fetoprotein (80% and 84.4%) and Alu methylation (66% and 61.5%). Furthermore, there was a strong positive correlation between Sat-α and Alu methylation. CONCLUSIONS: Measuring Sat-α and Alu methylation provides us with a new tool for early detecting HCV-induced CLD and hepatocarcinogenesis. Sat-α has the potential to be utilized as an independent predictive parameter for HCC development and progression because of its ability to distinguish between CLD and HCC with their different MELD scores.

Modulatory Effects of Arctostaphylos uva-urs Extract In Ovo Injected into Broiler Embryos Contaminated by Aflatoxin B1.

In ovo injection of nutrients can modulate the embryo's physiological responses against aflatoxin B1 (AFB1) embryotoxicity. This hypothesis was tested using in ovo injection of Arctostaphylos uva-ursi (Ar. uu.) methanolic extract. The total polyphenols, total flavonoids, total antioxidant capacity, and GC-MS analysis were all assessed in the Ar. uu. methanolic extract. A total of 180 ten-day-old embryonated eggs were distributed into six groups of 30 replicates each. The first group was used as a control (non-injected), and the second, third, fourth, fifth, and sixth groups were injected with 10 µ double-distilled water (DDW), 500 µL methanol, 0.01 g Ar. uu./500 µL methanol, 50 ng AFB1/10 µL DDW, and 50 ng AFB1 in 10 µ DDW + 0.01 g Ar. uu./500 µL methanol, respectively. The relative embryo weight, residual yolk sac weight, tibia length and weight, and survival were recorded. Total and differential leukocytes, oxidative stress, and humoral immune responses were observed. The residual yolk sac was lower (p < 0.05) in the Ar. uu. group than other groups. The embryonic growth (tibia weight and length) was enhanced in AFB1 + Ar. uu.-injected embryos compared with those injected with AFB1 alone. In conclusion, in ovo injection of Arctostaphylos uva-ursi could modulate AFB1-induced toxicity in chicken embryos.

Traumatic Brachial Plexus Palsy in Children: Long-Term Outcome and Strategy of Reconstruction.

BACKGROUND: Traumatic brachial plexus injuries in children represent a definite spectrum of injuries between adult and neonatal brachial plexus injuries. Their characteristics have been scarcely reported in the literature. The priority of functional restoration is not clear. METHODS: In total, 52 children with surgically treated traumatic brachial plexus injuries, excluding Erb's palsy, were reviewed after a minimum follow-up of 2 years. All children except nine were males, with an average age at surgery of 8 years. Forty-five children had exclusive supraclavicular plexus injuries. Twenty-one of them (46%) had two or more root avulsions. Seven children (13.5%) had infraclavicular plexus injuries. Time from trauma to surgery varied from 1 to 15 months (mean = 4.7 months). Extraplexal neurotization was the most common surgical technique used. RESULTS: Shoulder abduction and external rotation were restored to an average of 83 and 26 degrees, respectively. Elbow flexion and extension were restored to grade ≥3 in 96 and 91.5% of cases, respectively. Finger flexion and extension were restored to grade ≥4 in 29 and 32% of cases, respectively. Wrist flexion and extension were restored to grade ≥4 in 21 and 27% of cases, respectively. Results of neurotization were superior to those of neurolysis and nerve grafting. Among the 24 children with insensate hands, 20 (83.3%) recovered S3 sensation, 3 recovered S2, and 1 recovered S1. No case complained of neuropathic pain. Functional recovery correlated negatively but insignificantly with the age at surgery and time from injury to surgery. CONCLUSION: Brachial plexus injuries in children are associated with a high incidence root avulsions and no pain. Neurotization is frequently required and the outcome is not significantly affected by the delay in surgery. In total plexus injuries, some useful hand function can be restored, and management should follow that of obstetric palsy and be focused on innervating the medial cord.

Evaluation of the efficacy of commonly used disinfectants against isolated chlorine-resistant strains from drinking water used in Egyptian cattle farms.

BACKGROUND AND AIM: Drinking water of poor microbiological quality contains high percentages of microbes causing outbreaks of mainly coliform-related diseases. These microbes could be controlled by many hygienic standards including disinfection, but disinfectants misuse causes the developing of disinfectant-resistant strains. The present study aimed to investigate drinking water bacterial profile, determine chlorine-resistant strains, and statistically correlate that with the used disinfectant and disinfection process variables. In vitro evaluation of the bactericidal effect of the most commonly used disinfectants in cattle operations against the isolated chlorine-resistant strains and detection of qacE resistance gene in the isolated chlorine-resistant Escherichia coli strains in some cattle farms suffering coliform and non-coliform related disease around Egypt. MATERIALS AND METHODS: A structured questionnaire is used to survey a convenience sample of 132 Egyptian cattle beef and dairy farms suffering emerged epidemics to identify commonly used disinfection process, disinfectant types, disinfectants frequency, and rate of use. One hundred and thirty-two water samples were collected for microbiological analysis to obtain water bacterial profile and testing resistance to chlorine. Statistical analysis was performed to identify the level of association between microbial profile and presence of chlorine-resistant strains in each farm with used disinfection, disinfectant types, and rate of use in these farms. RESULTS: A wide range of disinfectant types used for variable purposes inside cattle farms with a different frequency of use and the highest percent of farms 25.8% use 4-5 types of disinfectants, followed by 25% of farms use two types, then 18.9% use three types. Microbial profile of water samples revealed isolation of E. coli, Streptococcus faecalis, Pseudomonas aeruginosa, Klebsiella spp., Proteus spp., Salmonella spp., Enterobacter spp., Citrobacter spp., Shigella flexneri, Serratia marcescens, and Yersinia enterocolitica in percent (98.5, 97.7, 97.7, 76.5, 66.7, 36.4, 78.8, 74.2, 30.3, 29.5, and 14.4% of cattle farms, respectively), from which five E. coli, four Salmonella, four Pseudomonas, two Klebsiella, and four Streptococcus strains expressed chlorine resistance. Statistical analysis showed weak to moderate correlation (rho 0.15-0.46) between bacterial profile strains count and presence of resistant strains with different farm disinfection, disinfectant types, and rate of use. Experimental evaluation of the bactericidal effect of the eight selected disinfectants on the chlorine-resistant isolated strains revealed that peroxymonosulfate killed 19/19 isolated strains/15 min contact time, and quaternary ammonium compounds killed only 3/19 strains/15 min contact time. The qacE resistance gene was detected in 3/4 isolated chlorine-resistant E. coli strains. CONCLUSION: Drinking water microbial profile strains and resistance to disinfectants are widely varied in cattle farms, and this variance depends on critical factors among which the disinfection process types used disinfectant types and frequency of disinfectants use or change.