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Posterior-stabilized, rotating platform knee prosthesis design was aimed to decrease polyethylene wear for the sake of improving implant survivorship. The purpose of the present prospective study was to evaluate the long-term clinical and radiographic results as well as the survival rate after using a rotating platform, posterior-stabilized knee prosthesis at a minimum of 10 years at a Middle East institution. We compared the results with reports in the literature on western populations. From January 2002 to June 2008, 96 patients (106 knees) underwent total knee arthroplasty (TKA) using a cemented rotating platform posterior-stabilized knee prosthesis. At a mean of 11.5 ± 1.3 years, 85 patients (95 TKAs) were available for clinical, radiographic, and implant survival analysis. At the final follow-up, 78.9% of the patients had excellent Knee Society Scoring system score, the average knee flexion was 110 ± 17 degrees, the average anatomical knee coronal alignment was 186 ± 2 degrees and 187 ± 3 degrees for varus and valgus knees, respectively. Five (5.2%) knees were revised of these: two for bearing dislocation, two for aseptic loosening, and one for infection. The Kaplan-Meier survival rate was 94.7% for all revisions and 97.8% when only revision for aseptic loosening considered as the end point. At a long-term follow-up, reasonable clinical and radiographic outcomes had been achieved after using a rotating platform, posterior-stabilized knee prosthesis in our population with acceptable survival rate reaching up to 95%, which is comparable to reports from the western population., Competing Interests: None declared., (Thieme. All rights reserved.)

Pseudomonas aeruginosa is a ubiquitous opportunistic bacterium that causes diseases in animals and humans. This study aimed to investigate the genetic diversity, antimicrobial resistance, biofilm formation, and virulence and antibiotic resistance genes of P. aeruginosa isolated from the uterus of cow, camel, and mare with clinical endometritis and their drinking water. Among the 180 uterine swabs and 90 drinking water samples analysed, 54 (20%) P. aeruginosa isolates were recovered. Isolates were identified biochemically to the genus level by the automated Vitek 2 system and genetically by the amplification of the gyrB gene and the sequencing of the 16S rRNA gene. Multilocus sequence typing identified ten different sequence types for the P. aeruginosa isolates. The identification of ST2012 was significantly ( p ≤ 0.05) higher than that of ST296, ST308, ST111, and ST241. The isolates exhibited significantly ( p ≤ 0.05) increased resistance to piperacillin (77.8%), ciprofloxacin (59.3%), gentamicin (50%), and ceftazidime (38.9%). Eight (14.8%) isolates showed resistance to imipenem; however, none of the isolates showed resistance to colistin. Multidrug resistance (MDR) was observed in 24 isolates (44.4%) with a multiple antibiotic resistance index ranging from 0.44 to 0.77. MDR was identified in 30 (33.3%) isolates. Furthermore, 38.8% and 9.2% of the isolates exhibited a positive extended-spectrum-β-lactamase (ESBL) and metallo-β-lactamase (MBL) phenotype, respectively. The most prevalent β-lactamase encoding genes were bla TEM and bla CTX-M , however, the bla IPM gene was not detected in any of the isolates. Biofilm formation was observed in 49 (90.7%) isolates classified as: 11.1% weak biofilm producers; 38.9% moderate biofilm producers; 40.7% strong biofilm producers. A positive correlation was observed between the MAR index and biofilm formation. In conclusion, the results highlighted that farm animals with clinical endometritis could act as a reservoir for MDR and virulent P. aeruginosa . The emergence of ESBLs and MBLs producing P. aeruginosa in different farm animals is a public health concern. Therefore, surveillance programs to monitor and control MDR P. aeruginosa in animals are required.

Johne's disease is a chronic infectious granulomatous enteritis caused by Mycobacterium avium subsp. paratuberculosis (MAP) and mainly infect wild and domestic animals. Although MAP infection has been reported worldwide, observational studies on MAP in camels are very scarce. The objective of the present investigation was to determine the herd- and camel-level seroprevalences and management factors associated with MAP seroprevalence in dromedary camels in the Eastern Province, Saudi Arabia. A cross-sectional study with two-stage random sampling was conducted. Serum samples from 391 camels in 67 herds were collected and tested for the presence of MAP antibodies using a commercial indirect ELISA test. The average MAP herd- and camel-level seroprevalences were 40.3% (95% CI: 29.10 - 52.60%) and 16.1% (95% CI: 12.78 - 20.11%), respectively. The herd-level factors showed a greater risk of MAP seropositivity in medium (36 - 75) and larger (>75) size herds compared with small (<36) herds. Furthermore, the risk of MAP seropositivity decreased in herds with calving pens compared to herds without calving pens. The camel-level factors indicated a decrease in seroprevalence of MAP with the age of camels. The present study revealed a high prevalence of MAP in dromedary camels in Eastern Province, Saudi Arabia. The herd-level risk factors for MAP seroprevalence identified in this study will provide the baseline data for developing and implementing a comprehensive control program for MAP in Saudi Arabia., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Backyard birds are small flocks that are more common in developing countries. They are used for poultry meat and egg production. However, they are also implicated in the maintenance and transmission of several zoonotic diseases, including multidrug-resistant bacteria. Enterococci are one of the most common zoonotic bacteria. They colonize numerous body sites and cause a wide range of serious nosocomial infections in humans. Therefore, the objective of the present study was to investigate the diversity in Enterococcus spp. in healthy birds and to determine the occurrence of multidrug resistance (MDR), multi-locus sequence types, and virulence genes and biofilm formation. From March 2019 to December 2020, cloacal swabs were collected from 15 healthy backyard broiler flocks. A total of 90 enterococci strains were recovered and classified according to the 16S rRNA sequence into Enterococcus faecalis (50%); Enterococcus faecium (33.33%), Enterococcus hirae (13.33%), and Enterococcus avium (3.33%). The isolates exhibited high resistance to tetracycline (55.6%), erythromycin (31.1%), and ampicillin (30%). However, all of the isolates were susceptible to linezolid. Multidrug resistance (MDR) was identified in 30 (33.3%) isolates. The enterococci AMR-associated genes ermB , ermA , tetM , tetL , vanA , cat, and pbp5 were identified in 24 (26.6%), 11 (12.2%), 39 (43.3%), 34 (37.7%), 1 (1.1%), 4 (4.4%), and 23 (25.5%) isolates, respectively. Of the 90 enterococci, 21 (23.3%), 27 (30%), and 36 (40%) isolates showed the presence of cylA , gelE, and agg virulence-associated genes, respectively. Seventy-three (81.1%) isolates exhibited biofilm formation. A statistically significant correlation was obtained for biofilm formation versus the MAR index and MDR. Multi-locus sequence typing (MLST) identified eleven and eight different STs for E. faecalis and E. faecium , respectively. Seven different rep-family plasmid genes (rep1-2, rep3, rep5-6, rep9, and rep11) were detected in the MDR enterococci. Two-thirds (20/30; 66.6%) of the enterococci were positive for one or two rep-families. In conclusion, the results show that healthy backyard chickens could act as a reservoir for MDR and virulent Enterococcus spp. Thus, an effective antimicrobial stewardship program and further studies using a One Health approach are required to investigate the role of backyard chickens as vectors for AMR transmission to humans.

The spread of SARS-CoV-2, the causative agent for COVID-19, has led to a global and deadly pandemic. To date, few drugs have been approved for treating SARS-CoV-2 infections. In this study, a structure-based approach was adopted using the SARS-CoV-2 main protease (M pro ) and a carefully selected dataset of 37,060 compounds comprising M pro and antiviral protein-specific libraries. The compounds passed two-step docking filtration, starting with standard precision (SP) followed by extra precision (XP) runs. Fourteen compounds with the highest XP docking scores were examined by 20 ns molecular dynamics simulations (MDs). Based on backbone route mean square deviations (RMSD) and molecular mechanics/generalized Born surface area (MM/GBSA) binding energy, four drugs were selected for comprehensive MDs analysis at 100 ns. Results indicated that birinapant, atazanavir, and ritonavir potently bound and stabilized SARS-CoV-2 M pro structure. Binding energies higher than -102 kcal/mol, RMSD values <0.22 nm, formation of several hydrogen bonds with M pro , favourable electrostatic contributions, and low radii of gyration were among the estimated factors contributing to the strength of the binding of these three compounds with M pro . The top two compounds, atazanavir and birinapant, were tested for their ability to prevent SARS-CoV-2 plaque formation. At 10 µM of birinapant concentration, antiviral tests against SARS-CoV-2 demonstrated a 37% reduction of virus multiplication. Antiviral assays demonstrated that birinapant has high anti-SARS-CoV-2 activity in the low micromolar range, with an IC50 value of 18 ± 3.6 µM. Therefore, birinapant is a candidate for further investigation to determine whether it is a feasible therapy option., Competing Interests: The authors declare there are no competing interests., (©2022 Kandeel et al.)

Background: Fasting Ramadan is one of the fundamental pillars of Islam. Although sick people are excluded from this duty, some of the hemodialysis patients insist to fast to enjoy the spiritual nature of the holy month., Objectives: To monitor the tolerability of fasting Ramadan among the hemodialysis population., Methodology: One hundred ninety-nine prevalent hemodialysis (HD) patients participated in the study and were allocated to 3 groups according to their fasting decision (complete, partial, and non- fasting). Basic demographic and laboratory data were collected before the start of the holy month; monitoring any inter or intradialytic complications or events during the holy month was done in addition to dry weight monitoring before and at the end of the month., Results: One hundred ninety-nine HD patients were included (97 males, mean age 45 ± 15 SD). Patients were divided based on their fasting state into 3 groups: compete fasting 28 (14%), partial fasting 88 (44%), and non-fasting 83 (42%). Out of 116 total fasting patients, only 4 patients (3.4%) developed complications (intradialytic hypotension (IDH) and muscle cramps) during dialysis. On the other hand, 3 patients experienced improvement of IDH; also, one patient reported improvement in dyspepsia. We noted a significant reduction in dry weight in the complete and partial fasting groups ( P  < 0.001 for both), unlike the non-fasting group ( P  = 0.75)., Conclusion: We may conclude that fasting Ramadan in hemodialysis patients whether complete or partial fasting may be tolerated by most of patients and was associated with a significant reduction in dry weight., Competing Interests: Competing interestsThe authors declare no competing interests., (© The Author(s) 2022.)

The present study aimed to determine the occurrence, genotypes, and antimicrobial resistance of Clostridium perfringens ( C. perfringens ) and Clostridioides difficile ( C. difficile ) in camel minced meat samples collected from small butcher shops and supermarkets in Al-Ahsa Governorate, Saudi Arabia. A total of 100 camel minced meat samples were randomly collected from small butcher's shops (n = 50) and supermarkets (n = 50) in Al-Ahsa Governorate, Saudi Arabia. C. perfringens and C. difficile were isolated and identified using the VITEK-2 compact system and 16S rRNA gene amplification. Genotypes, toxin genes, and antimicrobial susceptibility of the isolates were determined. Moreover, ELISA was used to detect C. perfringens and C. difficile toxins. C. perfringens and C. difficile were isolated from 14% and 4% of the tested minced meat samples, respectively. Out of the 14 C. perfringens isolates, type A (64.3%), type B (7.1%), type C (21.5%), and type D (7.1%) were detected. However, out of the four C. difficile isolates, three (75%) were type A + B + and one (25%) was type A - B + . None of the C. perfringens or C. difficile toxins were identified using ELISA. C. perfringens and C. difficile isolates exhibited a high rate of resistance to tetracycline (56% and 75%, respectively). However, all isolates were susceptible to amoxicillin-clavulanate. Multidrug resistance was observed in three (21.4%) C. perfringens and one (25%) C. difficile isolates. In conclusion, camel minced meat was contaminated with C. perfringens and C. difficile , which present a potential risk of food poisoning. The majority of the isolates were resistant to at least one antimicrobial, and some isolates were multidrug-resistant. Therefore, food safety standards and frequent inspections of abattoirs, small butcher shops, and supermarkets should be enforced.

Background and Aim: Backyard chicken flocks have traditionally been regarded as an essential food source in developed countries; however, they may act as reservoirs and spread various zoonotic bacterial pathogens. This study was designed to investigate the prevalence, phenotypic resistance, biofilm formation (BF), and pathotypes of Escherichia coli isolates from backyard poultry farms., Materials and Methods: Cloacal swabs (n=150) and internal organs (n=150) were collected from 30 backyard chicken flocks; 20 of them were experiencing systemic infection, and the other ten were apparently healthy. Samples were bacteriologically examined for E. coli isolation. Isolates were identified biochemically by the VITEK ® 2 COMPACT system (BioMérieux, France). For molecular identification, 16S rRNA was amplified and sequenced. Ten antimicrobials were selected for E. coli antimicrobial susceptibility testing. The minimum inhibitory concentration for each antimicrobial was determined. The extended-spectrum β-lactamase activity in isolates was investigated using cephalosporin/clavulanate combination disks. The ability of isolates for BF was determined by the microtiter plate method. Thirteen virulence genes linked to different E. coli pathotypes and two serotype-related genes were investigated by real-time polymerase chain reaction., Results: Eighty-six E. coli strains were isolated from 30 backyard chicken flocks. The isolates were biochemically identified to the species level. Genetically, sequences of the 16S rRN A gene showed >98% identity with E. coli in the National Center for Biological Information database. The frequency of isolation from diseased flocks was significantly higher (p<0.05) than apparently healthy flocks; 63.9% of the isolates were recovered from cloacal swabs and 36.04% were recovered from internal organs. E. coli isolates showed high resistance to ampicillin (AMP; 75.6%), gentamicin (39.5%), and tetracycline (29.1%). However, none of the isolates were resistant to imipenem. A variable drug resistance profile for E. coli isolates was reported. Twenty-one (24.4%) isolates were sensitive to all ten antimicrobials. Seven (8.1%) isolates were resistant only to AMP, and 28 (32.6%) were resistant to two antimicrobials, whereas the remaining 30 (34.9%) isolates showed multidrug resistance (MDR). Of the 86 isolates, 8 (9.3%) were confirmed as extended-spectrum β-lactamase (ESBL)-producing E. coli by the combination disk diffusion method. All ESBL isolates were MDR with an MDR index of 0.5-0.6. Fifty-seven (66.3%) isolates were capable of forming biofilms; 22 (25.6%) of them were strong biofilm producers, 24 (27.9%) moderate producers, and 11 (12.8%) weak producers. A statistically significant pairwise correlation was obtained for MDR versus BF (r=0.512) and MDR index versus BF (r=0.556). Based on virulence gene profiles, five pathotypes were identified, including enteropathogenic E. coli (39.5%), avian pathogenic E. coli (32.53%), enterohemorrhagic E. coli (EHEC; 9.3%), enterotoxigenic E. coli (ETEC; 5.8%), and enteroaggregative E. coli (EAEC; 1.2%). The lower frequency of EAEC and ETEC was statistically significant than other pathotypes. Three isolates were identified as O157 based on the detection of the rbfO157 gene., Conclusion: This study reported a high prevalence of MDR, suggesting the misuse of antimicrobials in backyard chicken farms. The emergence of ESBL and EHEC isolates in backyard chickens is a public health concern. Furthermore, the backyard flocks environment may harbor different pathogenic bacteria that may enhance the persistence of infection and the transmission to in-contact humans. Regular monitoring for the occurrence of MDR and the zoonotic pathotypes among E. coli in backyard chicken flocks is recommended, as these bacteria can transmit to humans through food products or contaminated environments., (Copyright: © Al-Marri, et al.)

Background: Intrastromal corneal ring segments are widely adopted for keratoconus management. However, the complete ring (Myoring) was proposed to be superior in advanced cases. Myoring can be implanted either via femtoassisted or manual dissection techniques. A comparison between both techniques can delineate any differences in the outcomes., Methods: This was a prospective interventional case series study. Sixty-four eyes with progressive advanced keratoconus were enrolled: 36 and 28 had femtoassisted or manual Myoring, respectively. Uncorrected visual acuity (UCVA), corrected distance visual acuity (CDVA), maximal keratometry ( K max ), spherical equivalent (SE) and corneal thinnest location were measured in all eyes preoperatively and at one, six, and 12 months postoperatively. Epi-off corneal cross-linking (CXL) was performed eight weeks after Myoring implantation for all cases., Results: Femtoassisted Myoring dissection significantly improved UCVA and CDVA from 0.1 ± 0.06 and 0.18 ± 0.1 preoperatively to 0.29 ± 0.08 and 0.43 ± 0.1 at 12 months. Also, manual technique similarly enhanced UCVA and CDVA from 0.11 ± 0.05 and 0.2 ± 0.1 preoperatively to 0.27 ± 0.2 and 0.4 ± 0.2 at 12 months. In terms of safety, while no cases of ring extrusion were encountered with the femtoassisted technique, six (21.4%) cases of extrusion were encountered in the manual group., Conclusion: Femtoassisted or manual Myoring technique followed by CXL is an effective choice for advanced progressive keratoconus. Although it did not reach a statistical significance, the high extrusion rate with manual dissection is a red flag to be considered., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2021 Ahmed Ibrahim Basiony et al.)

Q fever is a zoonotic disease caused by Coxiella burnetii ( C. burnetii ), an intracellular, Gram-negative bacterium that infects humans and domestic ruminants. Information on flock management factors associated with Q fever seropositivity in Saudi Arabia is very scarce. Therefore, the objective of this study was to identify the animal and flock management factors associated with Q fever seropositivity. For the assessment of risk factors, a case-control study was carried out. Cases ( n = 25) were flocks that had recent abortions within the previous two weeks and were PCR positive for C. burnetii . Control flocks ( n = 25) had no history of recent abortion and were PCR negative for C. burnetii . A questionnaire was developed to collect information about the flock management risk factors possibly associated with Q fever exposure in sheep. A total of 2437 sheep serum samples, collected from infected ( n = 1610, 10-150 samples/flock) and non-infected ( n = 827, 10-65 samples/flock) flocks, were tested for C. burnetii antibodies using a commercial ELISA kit between May 2018 and April 2019. In addition, 521 samples, including 50 aborted materials, 173 vaginal swabs, 134 faecal, and 164 milk samples, were collected for PCR testing. Infected flocks were 100% seropositive (within-flock seroprevalence ranging between 13.8% and 60%) and 100% PCR positive (with animal shedders of C. burnetii through aborted materials and/or vaginal fluids, feces, and milk). However, in non-infected control flocks, 28% were seropositive (within-flock seroprevalence ranging between 6.7% and 20%) and none had C. burnetii shedders. Epidemiological data were analyzed using mixed-effect logistic regression with a random effect for the flock. The results identified three protective factors: flocks with a lambing pen (odds ratio (OR): 0.46; 95% CI: 0.28-0.76), change bedding after removing aborted materials (OR: 0.42; 95% CI: 0.23-0.76), and flocks that isolated aborted ewes (OR: 0.41; 95% CI: 0.25-0.67), as well as two risk factors: flocks infested with ticks (OR: 2.78; 95% CI: 1.65-4.70) and flocks with a history of Q fever (OR: 3.03; 95% CI: 1.42-6.50). These results could be used to improve sheep flock biosecurity measures to prevent the introduction and reduce exposure of sheep and humans to Q fever infection.

Introduction: Hepatitis C virus (HCV) infection is an important cause of a variety of otherwise unexplained heart diseases and myocardial injury. A high prevalence of HCV infection has been noted in patients with hypertrophic cardiomyopathy, dilated cardiomyopathy, arrhythmogenic right ventricular dysplasia/cardiomyopathy and myocarditis. Various arrhythmias, conduction disturbances and QT prolongation were also associated with HCV infection. A possible role of HCV infection in the pathogenesis of diabetes and atherosclerosis, and the role of immunogenetics of HCV cardiomyopathies is discussed. Recent studies suggest that mononuclear cells may be the major target of HCV, and clinical applications to test this new hypothesis are discussed., Areas Covered: In this review, we will evaluate the evidence that HCV causes various cardiovascular diseases, and discuss on the pathogenesis of these disorders., Expert Opinion: HCV is the cause of many different forms of heart disease worldwide, but their existence has not been recognized by most of cardiologists. The recognition and diagnosis are indispensable for the early treatment of these diseases. The diverse clinical manifestation of HCV infection and the presence of multiple extrahepatic disease syndromes could be explained by a new hypothesis that the target of HCV is leukocytes.

Staphylococcal mastitis (SM) is a frequent disease in the dairy cattle that is costly to treat. This study aimed to investigate the alterations in the levels of procalcitonin (PCT), neopterin (NPT), haptoglobin (HP), serum amyloid A (SAA), proinflammatory cytokines (IL-1 β , IL-8, TNF- α , IF- γ ) and oxidative stress (OS) biomarkers in Holstein dairy cows with SM under field conditions. In addition, we also evaluated the role of examined biomarkers in disease pathogenesis and their use as diagnostic biomarkers for the disease in dairy cows. Fifty-three dairy cows with SM, including those with infections caused by Staphylococcus aureus ( n  = 42) and methicillin resistant S. aureus (MRSA) ( n  = 11) were selected for this study. In addition, 20 healthy dairy cows were enrolled as a control group. Higher serum levels of PCT, NP, IL-1 β , IL-8, TNF- α , IF- γ , HP and SAA and a state of OS was detected in SM group in comparison with the controls. Moreover, the levels of all examined biomarkers in mastitic cows with S. aureus when compared with those infected with MRSA was not significantly different. All examined biomarkers demonstrated a significant degree of discrimination between SM cows and healthy controls (the area under the curve (AUC) ranged from 83.6 for S AA to 100 for PCT). Our study showed that SM in dairy cows was associated with substantial changes in serum PCT, NPT, Acute phase proteins (APPs), proinflammatory cytokines, and OS levels. This study demonstrates that clinical examination in tandem with quantification of PCT, NPT, APPs and cytokines, OS biomarkers could be a useful assessment tool for SM in dairy cows., Competing Interests: The authors declare there are no competing interests., (©2021 El-Deeb et al.)

Chlamydia abortus ( C. abortus ) is intracellular, Gram-negative bacterium that cause enzootic abortion in sheep and goats. Information on C. abortus seroprevalence and flock management risk factors associated with C. abortus seropositivity in sheep and goats in Saudi Arabia are scarce. The objectives of this study were to (i) estimate the animal, flock, and within-flock seroprevalence of C. abortus among Eastern Province sheep and goat flocks and (ii) identify the flock management and animal risk factors associated with C. abortus seropositivity in Eastern Province, Saudi Arabia. A cross-sectional study with a two-stage sampling process was carried out in the Eastern Province, Saudi Arabia, between 2015 and 2016. A total of 1717 sheep and 1101 goat serum samples were collected from 21 sheep and 14 goat flocks, then were tested for C. abortus antibodies using a commercial ELISA Kit. In addition, vaginal swabs and aborted tissue samples were collected from sheep (n = 48) and goats (n = 15) with recent history of abortion for detection of C. abortus pmp gene using PCR. A questionnaire was constructed to collect information about flock management and animal risk factors possibly associated with C. abortus infection in sheep and goats. The true sheep and goat-level seroprevalences were 11.1% (95% CI: 9.7-12.7) and 10.6% (95% CI: 8.8-12.5), respectively. The true flock-level seroprevalence was 100% for both sheep and goats. However, the average within sheep and goat flocks true seroprevalences were 9.6% (95% CI: 1.8-22.9) and 9.3% (95% CI: 1.8-19.5), respectively. Multivariable logistic regression revealed that introduction of new sheep to the flocks (OR = 2.6; 95% CI: 1.5-4.4), type of breeding system (OR = 1.8; 95% CI: 1.0-3.4), flocks allowing females in (OR = 1.9; 95% CI: 1.1-3.3) or females out (OR = 2.2; 95% CI: 1.1-4.3), and sheep age 1.4-2.8 years (OR = 1.9; 95% CI: 1.3-2.9) were potential risk factors for C. abortus seropositivity in sheep flocks. However, in goat flocks, the introduction of new goats to the flocks (OR: 1.9; 95% CI: 1.2-3.0) was identified as a risk factor, whereas good farm hygiene (OR: 0.3; 95% CI: 0.2-0.7) was identified as a protective factor. C. abortus pmp gene was identified in 45 (93.8%) and 15 (100%) of samples collected from sheep and goats, respectively. These results could be used to implement efficient management measures to prevent and control C. abortus infection in sheep and goats in Eastern Province, Saudi Arabia, but also could be used to reduce the risk of C. abortus infection in sheep and goat flocks with similar management practices in other regions.

Understanding the distribution, antimicrobial resistance (AMR), and risk factors associated with multidrug-resistant (MDR) and methicillin-resistant staphylococci (MRS) isolated from cats admitted to veterinary clinics may decrease the risk of MDR and MRS transmission to humans and other cats. As such, the objectives of this study were to investigate the diversity in Staphylococcus spp. recovered from different anatomical locations in healthy and diseased cats and to determine the occurrence of MDR and MRS spp. as well as possible risk factors associated with colonization in these cats. Five swabs were collected from the anus, skin, ear canal, conjunctival sac, and nares of each cat (209 healthy and 191 diseased) admitted to a veterinary clinic in Eastern Province, Saudi Arabia, between January and December 2018. Prior to sample collection, cat owners completed a questionnaire collecting information on cat demographics, health status, management, and antimicrobial usage. In total, 179 Staphylococcus isolates were recovered from healthy ( n = 71) and diseased ( n = 108) cats, including 94 (52.5%) coagulase-positive staphylococci (CoPS), and 85 (47.5%) coagulase-negative staphylococci (CoNS). Five Staphylococcus spp. were identified, namely, Staphylococcus aureus , Staphylococcus pseudintermedius , Staphylococcus felis , Staphylococcus capitis , and Staphylococcus saprophyticus . Staphylococcus isolates were most commonly resistant to penicillin (56.4%) and ciprofloxacin (25.7%); however, no isolate was resistant to clindamycin. Thirty (16.8%) Staphylococcus spp. (24 S. aureus and 6 S. pseudintermedius ) isolates were MDR, with resistance to up to six different antibiotic classes. Only 17 (9.5%) Staphylococcus spp. (15 methicillin-resistant S. aureus and 2 methicillin-resistant S. pseudintermedius ) harbored the mecA gene. Risk factor analysis showed that cats with a history of antibiotic therapy, those raised mainly indoors with a child, and those who visit a veterinary clinic for treatment were at higher risk of MDR and MRS colonization. In conclusion, MDR and MRS were common in healthy and diseased cats in Saudi Arabia. Thus, an effective antimicrobial stewardship program and further studies using a One Health approach are required to investigate the role of cats as vectors for AMR transmission to humans.

Migratory wild birds acquire antimicrobial-resistant (AMR) bacteria from contaminated habitats and then act as reservoirs and potential spreaders of resistant elements through migration. However, the role of migratory wild birds as antimicrobial disseminators in the Arabian Peninsula desert, which represents a transit point for birds migrating all over Asia, Africa, and Europe not yet clear. Therefore, the present study objective was to determine antimicrobial-resistant bacteria in samples collected from migratory wild birds around Al-Asfar Lake, located in Al-Ahsa Oasis, Eastern Saudi Arabia, with a particular focus on Escherichia coli virulence and resistance genes. Cloacal swabs were collected from 210 migratory wild birds represent four species around Al-Asfar. E. coli , Staphylococcus , and Salmonella spp. have been recovered from 90 (42.9%), 37 (17.6%), and 5 (2.4%) birds, respectively. Out of them, 19 (14.4%) were a mixed infection. All samples were subjected to AMR phenotypic characterization, and results revealed (14-41%) and (16-54%) of E. coli and Staphylococcus spp. isolates were resistant to penicillins, sulfonamides, aminoglycoside, and tetracycline antibiotics. Multidrug-resistant (MDR) E. coli and Staphylococcus spp. were identified in 13 (14.4%) and 7 (18.9%) isolates, respectively. However, none of the Salmonella isolates were MDR. Of the 90 E. coli isolates, only 9 (10%) and 5 (5.6%) isolates showed the presence of eaeA and stx2 virulence-associated genes, respectively. However, both eaeA and stx2 genes were identified in four (4.4%) isolates. None of the E. coli isolates carried the hlyA and stx1 virulence-associated genes. The E. coli AMR associated genes bla CTX-M , bla TEM , bla SHV , aac(3)-IV , qnrA , and tet(A) were identified in 7 (7.8%), 5 (5.6%), 1 (1.1%), 8 (8.9%), 4 (4.4%), and 6 (6.7%) isolates, respectively. While the mecA gene was not detected in any of the Staphylococcus spp. isolates. Regarding migratory wild bird species, bacterial recovery, mixed infection, MDR, and AMR index were relatively higher in aquatic-associated species. Overall, the results showed that migratory wild birds around Al-Asfar Lake could act as a reservoir for AMR bacteria enabling them to have a potential role in maintaining, developing, and disseminating AMR bacteria. Furthermore, results highlight the importance of considering migratory wild birds when studying the ecology of AMR.

The papain-like protease (PL pro ) is an important enzyme for coronavirus polyprotein processing, as well as for virus-host immune suppression. Previous studies reveal that a molecular analysis of PL pro indicates the catalytic activity of viral PL pro and its interactions with ubiquitin. By using sequence comparisons, molecular models, and protein-protein interaction maps, PL pro was compared in the three recorded fatal CoV epidemics, which involved severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), severe acute respiratory syndrome CoV (SARS-CoV), and Middle East respiratory syndrome coronavirus (MERS-CoV). The pairwise sequence comparison of SARS-CoV-2 PL pro indicated similarity percentages of 82.59% and 30.06% with SARS-CoV PL pro and MERS-CoV PL pro , respectively. In comparison with SARS-CoV PL pro , in SARS-CoV-2, the PL pro had a conserved catalytic triad of C111, H278, and D293, with a slightly lower number of polar interface residues and of hydrogen bonds, a higher number of buried interface sizes, and a lower number of residues that interact with ubiquitin and PL pro . These features might contribute to a similar or slightly lower level of deubiquitinating activity in SARS-CoV-2 PLpro. It was, however, a much higher level compared to MERS-CoV, which contained amino acid mutations and a low number of polar interfaces. SARS-CoV-2 PL pro and SARS-CoV PL pro showed almost the same catalytic site profiles, interface area compositions and polarities, suggesting a general similarity in deubiquitination activity. Compared with MERS-CoV, SARS-CoV-2 had a higher potential for binding interactions with ubiquitin. These estimated parameters contribute to the knowledge gap in understanding how the new virus interacts with the immune system., (© 2020 Wiley Periodicals LLC.)

The objectives of the present study were to characterize Mycobacterium avium subsp. paratuberculosis (MAP) infection using serological and molecular tools and investigate the distribution and molecular characterization of MAP strains (cattle (C) and sheep (S) types) in sheep, goat, cattle, and camel herds in Eastern Province, Saudi Arabia. Serum and fecal samples were collected from all animals aged >2 years old in 31 herds (sheep = 8, goats = 6, cattle = 8 and camels = 9) from January to December 2019. Serum samples were tested by ELISA for the detection of MAP antibodies. Fecal samples were tested by PCR for the detection of MAP IS 900 gene and the identification of MAP strains. MAP antibodies were detected in 19 (61.3%) herds. At the animal level, antibodies against MAP were detected in 43 (19.5%) sheep, 21 (17.1%) goats, 13 (19.7%) cattle and 22 (9.1%) camels. The IS 900 gene of MAP was detected in 23 (74.2%) herds and was directly amplified from fecal samples of 59 (26.8%) sheep, 34 (27.6%) goats, 20 (30.3%) cattle and 36 (15.0%) camels. The S-type was the most prevalent MAP type identified in 15 herds, and all were identified as type-I, while the C-type was identified in only 8 herds. The IS 900 sequences revealed genetic differences among the MAP isolates recovered from sheep, goats, cattle and camels. Results from the present study show that MAP was prevalent and confirm the distribution of different MAP strains in sheep, goat, cattle and camel herds in Eastern Province, Saudi Arabia.

Clostridium difficile (CD) is considered a major health care problem both in developing and developed countries; frequently reported to be associated with enterocolitis and diarrhea in horses and other animals. In this study, we examined acute phase response (APR), cytokines response, neopterin (NP) procalcitonin (PCT) production and oxidative stress condition in horses and foals with C. difficile-induced enterocolitis (CDIE) and evaluated the effectiveness of these parameters as biomarkers for the disease. A total of 407 Arabian horses in 35 stables were examined between January 2017 to December 2018. Only 24 out of 407 horses showed two or more signs of CDIE. The blood level of serum amyloid A (SAA), haptoglobin (HP), proinflammatory cytokines (TNF-α, IL-6 and IL1-β), serum malondialdehyde (MDA), PCT and NPT in horses with CDIE were higher than in healthy horses. Nevertheless, the levels of nitric oxide (NO), superoxide dismutase (SOD) and total antioxidant concentration (TAC) were considerably lower in diseased horses compared to those that were healthy. The ROC curves for eleven selected blood parameters, both in healthy horses and horses with CDIE demonstrated that all examined blood markers had significant levels of differentiation between CDIE cases and healthy controls (AUC > 87.5). The data in this study suggest that the evaluation of acute-phase proteins, cytokines, PCT, NPT, and oxidative stress biomarkers may well be used as a tool for diagnosis and assessment of CDIE and in disease pathogenesis in Arabian horses., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Acute-phase proteins (APPs) have always had valued diagnostic potentialities in response to infection. This study aimed to evaluate the diagnostic accuracy of selected APPs and proinflammatory cytokines (PIC) in goats with contagious caprine pleuropneumonia (CCPP) under field conditions. Moreover, to highlight the role of tested biomarkers in CCPP pathogenesis. Fifty-eight goats (38 confirmed cases with CCPP and 20 healthy controls) were involved in this investigation. C-reactive protein (CRP), procalcitonin (PCT), haptoglobin (HP), fibrinogen (Fb), serum amyloid A (SAA), selected PIC (IL1- α , IL1- β , IL-6, interferon-gamma (IFN- γ ) and tumor necrosis factor-alpha (TNF- α )) levels were investigated in serum samples from all goats under investigation. Latex agglutination test was used for diagnosis of goats with CCPP. For microbiological investigations, nasopharyngeal swabs (from all goats), lung tissues and pleural fluids (from only necropsied goats) were collected. This study revealed that all tested parameters have a high to moderate degree of diagnostic performance for CCPP. Magnitudes of increase in levels of APPs (CRP, HP and SAA) were stronger than PIC, IFN- γ , Fb and PCT. All tested parameters showed high diagnostic accuracy (AUROC >90%), except HP (AUROC = 87.3%) and IFN- γ (AUROC = 78.8%) showed moderate accuracy in differentiation of goats with and without CCPP infection. For detecting goats with and without CCPP infection, HP had the lowest sensitivity (Se = 81.6%) and Fb had the lowest specificity (Sp = 85.0%) among the APPs parameters tested. However, PCT showed the highest Se (100%) and Sp (95.0%) to detect goats with and without CCPP infection among tested parameters. Conclusively, this study endorses the significance of selected APPs and PIC as additional screening diagnostic parameters for naturally occurring CCPP in goats. However, it does not replace traditional methods for diagnosis of CCPP in goats. Furthermore, APPs and PIC have an important role in disease pathogenesis in goats., Competing Interests: The authors declare there are no competing interests., (©2020 El-Deeb et al.)

Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an emerging disease with fatal outcomes. In this study, a fundamental knowledge gap question is to be resolved by evaluating the differences in biological and pathogenic aspects of SARS-CoV-2 and the changes in SARS-CoV-2 in comparison with the two prior major COV epidemics, SARS and Middle East respiratory syndrome (MERS) coronaviruses., Methods: The genome composition, nucleotide analysis, codon usage indices, relative synonymous codons usage, and effective number of codons (ENc) were analyzed in the four structural genes; Spike (S), Envelope (E), membrane (M), and Nucleocapsid (N) genes, and two of the most important nonstructural genes comprising RNA-dependent RNA polymerase and main protease (Mpro) of SARS-CoV-2, Beta-CoV from pangolins, bat SARS, MERS, and SARS CoVs., Results: SARS-CoV-2 prefers pyrimidine rich codons to purines. Most high-frequency codons were ending with A or T, while the low frequency and rare codons were ending with G or C. SARS-CoV-2 structural proteins showed 5 to 20 lower ENc values, compared with SARS, bat SARS, and MERS CoVs. This implies higher codon bias and higher gene expression efficiency of SARS-CoV-2 structural proteins. SARS-CoV-2 encoded the highest number of over-biased and negatively biased codons. Pangolin Beta-CoV showed little differences with SARS-CoV-2 ENc values, compared with SARS, bat SARS, and MERS CoV., Conclusion: Extreme bias and lower ENc values of SARS-CoV-2, especially in Spike, Envelope, and Mpro genes, are suggestive for higher gene expression efficiency, compared with SARS, bat SARS, and MERS CoVs., (© 2020 Wiley Periodicals, Inc.)

The present study aimed to isolate and genotype C. perfringens from healthy and diarrheic dromedary camels, pastures and herders; and to evaluate and compare antimicrobial susceptibility of the isolates. A total of 262 (56.3%) C. perfringens isolates were recovered from 465 samples of healthy and diarrheic dromedary camels, pastures and herders. C. perfringens type A (75.2%), type B (4.2%), type C (13.7%) and type D (6.9%) were detected. C. perfringens type A with only cpa + gene was found in 191 (72.9%) isolates and with cpa + associated cpb2 + was found only in 6 (2.3%) isolates. None of the isolates were positive for cpe and iap genes. The highest antimicrobial resistance (82.8%) was observed to ceftiofur with MIC 50 and MIC 90 values of <64 and ≥256 μg/mL, respectively, followed by penicillin G (72.9%) and erythromycin (61.5%). The lowest resistance (1.9%) was observed for doxycycline with MIC 50 and MIC 90 values of <1 and 4 μg/mL, respectively, followed by florfenicol (5.3%) and clindamycin (12.2%). In conclusion, C. perfringens type A with cpa + gene was the most prevalent toxin type isolated in this study. The majority of the isolates were resistant to at least one of the ten antimicrobials tested. Antimicrobial resistance patterns of C. perfringens isolates provide further evidence on the emergence of multiple-drug resistant C. perfringens. Therefore, the dissemination of surveillance programs to monitor and control C. perfringens in dromedary camels is required., Competing Interests: Declaration of Competing Interest No conflict of interests regarding the publication of this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

High-throughput cultivation methods have recently been developed to accelerate the recovery of microorganisms reluctant to cultivation. They simulate in situ environmental conditions for the isolation of environmental microbiota through the exchange of growth substrates during cultivation. Here, we introduce leaf-based culture media adopting the concept of the plant being the master architect of the composition of its microbial community. Pre-physical treatments of sunflower plant leaves, namely punching, freezing, and/or autoclavation, allowed the diffusion of electrolytes and other nutrients to configure the leaf surface as a natural pad, i.e., creating an " in situ similis " environment suitable for the growth of rarely isolated microbiota. We used surface inoculation and membrane-filtration methods to assess the culturability of endophytic bacteria from the sunflower phyllosphere and rhizosphere. Both methods supported excellent colony-forming unit (CFU) development when compared to standard R2A medium, with a special affinity to support better growth of epiphytic and endophytic populations of the phyllosphere compared with the rhizosphere. A 16S rRNA gene analysis of >122 representative isolates indicated the cultivation of a diverse set of microorganisms by application of the new methods. It indicated the predominance of 13 genera of >30 potential species, belonging to Firmicutes, Proteobacteria, and Actinobacteria, and especially genera not commonly reported for sunflower, e.g., Rhizobium , Aureimonas , Sphingomonas , Paracoccus , Stenotrophomonas , Pantoea , Kosakonia , and Erwinia . The strategy successfully extended diversity and richness in the endophyllosphere compared to the endorhizosphere, while CFUs grown on the standard R2A medium mainly pertain to Firmicutes, especially Bacillus spp. MALDI-TOF MS analysis clustered the isolates according to their niche and potential functions, where the majority of isolates of the endorhizosphere were clustered away from those of the endophyllosphere. Isolates identified as Gammaproteobacteria and Alphaproteobacteria were distinguishably sub-clustered, which was in contrast to the heterogeneous isolates of Firmicutes ( Bacillus spp.). In conclusion, leaf in situ similis cultivation is an effective strategy to support the future application of culturomics of plant microbiota. This is an effort to access novel isolates that are more adapted and competitive in their natural environments, especially those subjected to abiotic stresses like those prevailing in arid/semi-arid zones, and, consequently, to support the application of agro-biotechnologies, among other technologies, to improving agriculture in such zones., (Copyright © 2020 Nemr, Khalil, Sarhan, Abbas, Elsawey, Youssef, Hamza, Morsi, El-Tahan, Fayez, Patz, Witzel, Ruppel, El-Sahhar and Hegazi.)

Bovine respiratory diseases (BRD) have long been considered a serious problem that causes major economic losses in feedlot calves (FC). This study aimed to determine the diagnostic and prognostic effect of selected biological markers including, procalcitonin (PCT), neopterin (NP), proinflammatory cytokines (IL-1β, IL-8, TNF-α, IF-γ), haptoglobin (HP) and serum amyloid A (SAA) on FC with BRD under field conditions. Sixty-nine FC that were identified to be infected with Mannheimia haemolytica and Histophilus somni and had different clinical respiratory signs (diseased group) were selected for this study. In addition, 20 healthy FC have been selected as a control group. We have detected higher serum levels of PCT, NP, HP, SAA, IL-1β, IL-8, TNF-α and IF-γ in diseased FC group compared with the control group. All tested markers revealed a high level of discrimination between BRD infected FC and healthy ones (AUC > 0.90). Moreover, the obtained data showed a high degree of prognostic accuracy for PCT, NP, IL-8, HP, IF-γ and IL-1β in predicting treatment response of FC with BRD at the selected thresholds (AUC = 0.99, 0.99, 0.97, 0.93, 0.88 and 0.82, respectively). Significant inhibition was observed for the selected biochemical markers in treated FC 7 days post-treatment. In conclusion, this study showed that BRD in FC was associated with significant alterations in serum APPs, proinflammatory cytokines, PCT and NPT levels. Furthermore, it demonstrated that these serum biomarkers are much higher in FC with BRD compared to recovered ones. Our data suggest that the measurement of PCT, NPT, APPs and cytokines together with the clinical examination may be a useful diagnostic and prognostic tool for assessment of FC naturally infected with M. haemolytica and H. somni., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Lactobacillus spp. is one of the beneficial lactic acid producing microbiota in the vagina, which is important for a healthy vaginal environment. However, little is known about vaginal Lactobacillus in dromedary camels ( Camelus dromedarius ). Therefore, this study aimed to isolate vaginal lactic acid bacteria (LAB) in dromedary camels and to study the probiotic potential of selected isolates. A total of 75 vaginal swabs were collected from pluriparous, non-pregnant, non-lactating dromedary camels. The LAB were isolated using deMan, Rogosa and Sharpe broth and agar media. Suspected LAB isolates were subjected to catalase testing and Gram staining and examined for indole production, nitrate reduction, hemolytic activity, cell surface hydrophobicity, auto- and coaggregation, antibacterial activity and characterized by 16S rRNA amplification and sequencing. Eighteen LABs were isolated from the 75 vaginal swabs. Among the 18 LAB isolates, six were Lactobacillus plantarum , eight were Lactobacillus fermentum , and four were Lactobacillus rhamnosus . None of the LAB isolates was hemolytic and only four LAB were H 2 O 2 producing. The percentage of hydrophobicity ranged from 0% to 49.6%, 0% to 44.3% and 0% to 41.6% for hexadecane, xylene and toluene, respectively. All isolates showed higher ( P < 0.05) autoaggregation after 24 h of incubation compared to 4 h. Furthermore, all LAB showed higher coaggregation ( P < 0.05) and antimicrobial activity toward Staphylococcus aureus than to Escherichia coli . All LAB isolates were vancomycin resistant and sensitive to streptomycin, erythromycin, kanamycin and chloramphenicol. Only, three LAB isolates were resistant to tetracycline. The dromedary camel vaginal LAB isolates exhibited varying degrees of in vitro probiotic properties tested in this study and showed promising activity against the most common bacterial causes of endometritis in dromedary camels. Further investigation of the in vivo effect of these isolates is warranted., Competing Interests: The authors declare that they have no competing interests., (© 2020 El-Deeb et al.)

The recent introduction of plant-only-based culture media enabled cultivation of not-yet-cultured bacteria that exceed 90% of the plant microbiota communities. Here, we further prove the competence and challenge of such culture media, and further introduce "the inoculum-dependent culturing strategy, IDC". The strategy depends on direct inoculating plant serial dilutions onto plain water agar plates, allowing bacteria to grow only on the expense of natural nutrients contained in the administered inoculum. Developed colonies are successively transferred/subcultured onto plant-only-based culture media, which contains natural nutrients very much alike to those found in the prepared plant inocula. Because of its simplicity, the method is recommended as a powerful tool in screening programs that require microbial isolation from a large number of diverse plants. Here, the method comfortably and successfully recovered several isolates of endophytic Actinobacteria represented by the six genera of Curtobacterium spp., Plantibacter spp., Agreia spp., Herbiconiux spp., Rhodococcus spp., and Nocardioides spp. Furthermore, two of the isolates are most likely novel species belonging to Agreia spp. and Herbiconiux spp.

Acute phase proteins (APPs) and oxidative stress are helpful markers in diagnosis of several infectious diseases. APPs, proinflammatory cytokines and oxidative stress markers were evaluated for their role in the diagnosis of naturally acquired Coxiella burnetii (Q fever) associated with abortion in sheep, goats and she-camels. Blood, aborted materials and vaginal swabs were collected from mixed herds in the Eastern Province of Saudi Arabia. Antioxidant biomarkers showed significant decline in cases of abortion compared to control animals at delivery time. The correlation between disease status and all parameters ranged from moderate to high. The APPs, cytokines and the oxidative stress marker malondialdehyde (MDA) displayed a high degree of distinction between aborted sheep and goat and normal delivered animals (AUC > 0.90). However, only MDA showed a high degree of differentiation (AUC > 0.90) between aborted she-camels and normal delivered controls. In conclusion, results from our study allow us to recommend using APPs, cytokines and oxidative stress markers as an additional tool for diagnosis of naturally occurring C. burnetii infection in sheep, goats and she-camels. However, it does not replace standard procedures for detection of C. burnetii., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Introduction: Few centers worldwide have advanced single-incision pediatric endosurgery (SIPES) for pediatric age group. Up to our knowledge this is the first study assessing the safety of SIPES cholecystectomy in children with sickle cell disease (SCD) done by surgical residents. Aims: To determine the feasibility, safety, and expediency of SIPES cholecystectomy in children with SCD performed by surgical residents. Materials and Methods: Retrospective study of all SIPES cholecystectomies performed in our unit from April 1, 2011 to March 31, 2018. We compared the outcome of SIPES cholecystectomy done by fellows and residents. SIPES Covidien 5-12 mm port was inserted through umbilicus. Long 50 cm laparoscope, straight regular instruments, and transabdominal gallbladder traction suture were used in all patients. The cystic duct and artery were identified and divided. The gallbladder is then dissected off the liver and extracted from the abdomen through the port. Results: Forty-three SCD patients underwent SIPES cholecystectomy for 7 years. Mean HbS was 37.56%. Mean age was 10 years. Twenty-four cases (56%) and 19 (44%) were performed by 8 fellows and 10 residents, respectively. Nine other procedures were done simultaneously with cholecystectomy and were excluded from the mean operative time (MOT). The MOT for all cases was 85 minutes, 78 minutes for fellows and 94 minutes for residents ( P value is <.001). One extra port was inserted in 2 patients at the beginning of the series. Conclusion: SIPES cholecystectomy in children with SCD is safe and feasible if done by residents under supervision. Stepwise training is essential in this challenging surgery. Involving residents with other SIPES procedures and hands-on minimally invasive surgery single port workshops help them in improving their skills.

Aim: Egypt is the habitat for a large number of bird species and serves as a vital stopover for millions of migratory birds during their annual migration between the Palearctic and Afrotropical ecozones. Surveillance for avian influenza viruses (AIVs) is critical to assessing risks for potential spreading of these viruses among domestic poultry. Surveillance for AIV among hunted and captured wild birds in Egypt was conducted in order to understand the characteristics of circulating viruses., Methods: Sampling of wild bird species occurred in two locations along the Mediterranean Coast of Egypt in the period from 2014 to 2016. A total of 1316 samples (cloacal and oropharyngeal swabs) were collected from 20 different species of hunted or captured resident and migratory birds sold at live bird markets. Viruses were propagated then sequenced. Phylogenetic analysis and receptor binding affinities were studied., Results: Eighteen AIVs (1.37%) were isolated from migratory Anseriformes at live bird markets. Further characterization of the viral isolates identified five hemagglutinin (H3, H5, H7, H9, and H10) and five neuraminidase (N1, N2, N3, N6, and N9) subtypes, which were related to isolates reported in the Eurasian region. Two of the 18 isolates were highly pathogenic H5N1 viruses related to clade 2.2.1, while three isolates were G1-like H9N2 viruses., Conclusions: Our data show significant diversity of AIVs in Anserifromes sold at live bird markets in Egypt. This allows for genetic exchanges between imported and enzootic viruses and put the exposed humans at a higher risk of infection., (© 2019 The Authors. Influenza and Other Respiratory Viruses Published by John Wiley & Sons Ltd.)

Improving cultivability of a wider range of bacterial and archaeal community members, living natively in natural environments and within plants, is a prerequisite to better understanding plant-microbiota interactions and their functions in such very complex systems. Sequencing, assembling, and annotation of pure microbial strain genomes provide higher quality data compared to environmental metagenome analyses, and can substantially improve gene and protein database information. Despite the comprehensive knowledge which already was gained using metagenomic and metatranscriptomic methods, there still exists a big gap in understanding in vivo microbial gene functioning in planta , since many differentially expressed genes or gene families are not yet annotated. Here, the progress in culturing procedures for plant microbiota depending on plant-based culture media, and their proficiency in obtaining single prokaryotic isolates of novel and rapidly increasing candidate phyla are reviewed. As well, the great success of culturomics of the human microbiota is considered with the main objective of encouraging microbiologists to continue minimizing the gap between the microbial richness in nature and the number of species in culture, for the benefit of both basic and applied microbiology. The clear message to fellow plant microbiologists is to apply plant-tailored culturomic techniques that might open up novel procedures to obtain not-yet-cultured organisms and extend the known plant microbiota repertoire to unprecedented levels.

Introduction: Few centers worldwide have advanced single-incision pediatric endosurgery (SIPES) splenectomy. The safety and feasibility of SIPES performed by trainees were not assessed before. SIPES splenectomy is a demanding technique that needs high level of skills. This is the largest series of 39 SIPES splenectomies performed by 14 trainees in one single center., Aims: To assess the safety, feasibility, and technical challenges of SIPES splenectomy performed by trainees and to compare it with other published series., Materials and Methods: Retrospective study of all patients operated with SIPES splenectomy for 7 years. SIPES port was inserted through 1.5 cm umbilical incision. One type of port and straight regular instruments were used in all cases., Results: Forty-nine patients underwent SIPES splenectomy. Eighty percent was done by our trainees. Six cholecystectomies were done simultaneously. Forty-five patients with sickle cell disease, two with thalassemia, one spherocytosis, and one Fanconi's anemia. Mean operative time (MOT) for splenectomy was 182 minutes (130-190) and 251 minutes for splenectomy with cholecystectomy (230-270) depending on severity of adhesions and size of the spleen; P value <.001. Two conversions have to be opened due to bleeding. There is neither wound infection nor incisional hernia up to date., Conclusion: SIPES splenectomy is safe and feasible when performed by surgical trainees without adding any morbidity to the patient. Learning curve can quickly improve with more exposure of trainee to different SIPES procedures and hands-on workshops. More than one procedure could be done at the same time. It has excellent cosmesis and almost invisible scar.

The rapid development of high-throughput techniques and expansion of bacterial databases have accelerated efforts to bring plant microbiomes into cultivation. We introduced plant-only-based culture media as a successful candidate to mimic the nutritional matrices of plant roots. We herein employed a G3 PhyloChip microarray to meticulously characterize the culture-dependent and -independent bacterial communities of the maize root compartments, the endo- and ecto-rhizospheres. An emphasis was placed on the preference of the growth of unculturable candidate divisions/phyla on plant-only-based culture media over standard culture media (nutrient agar). A total of 1,818 different operational taxonomic units (OTUs) were resolved representing 67 bacterial phyla. Plant-only-based culture media displayed particular affinity towards recovering endophytic over ectophytic rhizobacteria. This was shown by the slightly higher recovery of CFUs for endophytes on plant-only-based culture media (26%) than on standard culture media (10%) as well as the higher taxa richness and numbers of exclusive families of unculturable divisions/phyla. Out of 30 bacterial phyla (comprising >95% of the whole population), 13 were of a significantly higher incidence on plant-only-based culture media, 6 phyla of which were not-yet-cultured (Atribacteria, OP9; Dependentiae, TM6; Latescibacteria, WS3; Marinimicrobia, SAR406; Omnitrophica, OP3; BRC1). Furthermore, plant-only-based culture media significantly enriched less abundant and/or hard-to-culture bacterial phyla (Acidobacteria, Gemmatimonadetes, and Tenericutes). These results present conclusive evidence of the ability of plant-only-based culture media to bring the plant-fed in situ microbiome into the status of plant-fed in vitro cultures, and to widen the scope of cultivation of heretofore-unculturable bacterial divisions/phyla.

A cross sectional study was conducted on 1010 goats from 25 flocks located in the eastern province, Saudi Arabia, to study the prevalence of methicillin resistant Staphylococcus aureus (MRSA) among goat farms. A total of 235 milk samples and 775 nasal swabs were collected for bacteriological investigation. Based on resistance to cefoxitin, 20 isolates were permissively identified as MRSA. PCR with specific primers was used to confirm MRSA. The prevalence of MRSA was 2%; with maximum prevalence in mastitic milk (9.2%) and swabs from animals showed respiratory signs (2.6%), while the lowest prevalence was identified in apparently normal milk (0.6). The standard disk diffusion test was used for in vitro evaluation of isolates resistance profile to 13 antimicrobial agents. Multidrug resistance (MDR) was detected in all MRSA and in 23.5% of methicillin sensitive Staphylococcus aureus (MSSA). Univariable association between the prevalence of MDR/MRSA strains and management practices indicated a higher prevalence with larger size flocks, where raising animals for both meat and milk production, and where antibiotics were used during the last 30 days, the latter was particularly pertinent to penicillin-streptomycin. Multivariable models indicated that larger flocks (200-400, and >400) were, respectively, 4 and 3.5 fold more likely to have MDR S. aureus compared to smaller flocks (<200)., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Habronemosis is a common parasitic disease of horses worldwide. In order to investigate how haptoglobin (Hp), serum amyloid A (SAA), oxidative stress markers, nitric oxide (NO), interleukin-6 (IL-6) and tumor necrosis factor (TNF- α), varies in cutaneous habronemosis, 30 horses with the clinical disease and 20 clinically healthy horses were included in the current study. The serum levels of Hp, SAA, and proinflammatory cytokines (IL-6 and TNF-α), NO, malondialdehyde (MDA), super oxide dismutase (SOD), glutathione (GSH), and total antioxidant capacity (TAC) were determined in horses before and after two weeks of treatment. The serum levels of Hp, SAA, IL-6, TNF-α and MDA were significantly elevated in infected horses as compared to the controls. Alternately, the serum levels of SOD, GSH, TAC and NO, were recorded low in infected horses as compared to the controls. All tested markers resumed the same levels after treatment as in control group. The Hp, SAA, IL-6, TNF-α, and MDA exhibited a high degree of clinical accuracy of the cases diagnosis. The area under the curve (AUC) for acute phase proteins (SAA, Hp), IL-6, TNF-α, and MDA was 0.87, 0.94, 0.96, 0.96 and 1.0, respectively. These findings showed that Hp, SAA, IL-6, TNF-α, and MDA may be supportive in the diagnosis of cutaneous habronemosis in horses and, simultaneously, they can also be used to monitor the progress of the treatment., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In order to improve the culturability and biomass production of rhizobacteria, we previously introduced plant-only-based culture media. We herein attempted to widen the scope of plant materials suitable for the preparation of plant-only-based culture media. We chemically analyzed the refuse of turfgrass, cactus, and clover. They were sufficiently rich to support good in vitro growth by rhizobacteria isolates representing Proteobacteria and Firmicutes. They were also adequate and efficient to produce a cell biomass in liquid batch cultures. These culture media were as sufficient as artificial culture media for the cultivation and recovery of the in situ rhizobacteria of barley (Hordeum murinum L.). Based on culture-dependent (CFU plate counting) and culture-independent analyses (qPCR), mowed turfgrass, in particular, supported the highest culturable population of barley endophytes, representing >16% of the total bacterial number quantified with qPCR. This accurately reflected the endophytic community composition, in terms of diversity indices (S', H', and D') based on PCR-DGGE, and clustered the plant culture media together with the qPCR root populations away from the artificial culture media. Despite the promiscuous nature of the plant materials tested to culture the plant microbiome, our results indicated that plant materials of a homologous nature to the tested host plant, at least at the family level, and/or of the same environment were more likely to be selected. Plant-only-based culture media require further refinements in order to provide selectivity for the in vitro growth of members of the plant microbiome, particularly difficult-to-culture bacteria. This will provide insights into their hidden roles in the environment and support future culturomic studies.

The plant-based-sea water culture medium is introduced to in vitro cultivation and in situ recovery of the microbiome of halophytes. The ice plant ( Mesembryanthemum crystallinum ) was used, in the form of juice and/or dehydrated plant powder packed in teabags, to supplement the natural sea water. The resulting culture medium enjoys the combinations of plant materials as rich source of nutrients and sea water exercising the required salt stress. As such without any supplements, the culture medium was sufficient and efficient to support very good in vitro growth of halotolerant bacteria. It was also capable to recover their in situ culturable populations in the phyllosphere, ecto-rhizosphere and endo-rhizosphere of halophytes prevailing in Lake Mariout, Egypt. When related to the total bacterial numbers measured for Suaeda pruinosa roots by quantitative-PCR, the proposed culture medium increased culturability (15.3-19.5%) compared to the conventional chemically-synthetic culture medium supplemented with (11.2%) or without (3.8%) NaCl. Based on 16S rRNA gene sequencing, representative isolates of halotolerant bacteria prevailed on such culture medium were closely related to Bacillus spp., Halomonas spp., and Kocuria spp. Seed germination tests on 25-50% sea water agar indicated positive interaction of such bacterial isolates with the germination and seedlings' growth of barley seeds.

In an effort to axenically culture the previously uncultivable populations of the rhizobacteria of Lucerne (Medicago sativa L.), we propose plant-only teabags culture media to mimic the nutritional matrix available in the rhizosphere. Here, we show that culture media prepared from Lucerne powder teabags substantially increased the cultivability of Lucerne rhizobacteria compared with a standard nutrient agar, where we found that the cultivable populations significantly increased by up to 60% of the total bacterial numbers as estimated by Quantitative Real-time Polymerase Chain Reaction (qRT-PCR). Cluster analysis of 16S rDNA Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) of cultivable Colony-Forming Units (CFUs) revealed a more distinct composition and separation of bacterial populations recovered on the plant-only teabags culture media than those developed on a standard nutrient agar. Further, the new plant medium gave preference to the micro-symbiont Sinorhizobium meliloti, and succeeded in isolating a number of not-yet-cultured bacteria, most closely matched to Novosphingobium sp., Lysobacter sp. and Pedobacter sp. The present study may encourage other researchers to consider moving from the well-established standard culture media to the challenging new plant-only culture media. Such a move may reveal previously hidden members of rhizobacteria, and help to further explore their potential environmental impacts.

Classical contagious caprine pleuropneumonia is one of the most fatal contagious disease of goats listed by World Organization for Animal Health that leads to major economic losses. It is caused by infection with Mycoplasma capricolum subspecies capripneumoniae. In order to isolate the causative agents of CCPP for the first time in the Kingdom of Saudi Arabia, fifteen flocks from Eastern region (Al Ahsa, Dammam and Hafr Albaten) and ten flocks from Riyadh and Al-Kharj regions were selected for this study. A total of 700 samples (400 nasal swabs, 300 pleural fluid samples and lung samples (from necropsied animals)) were collected from goats showing typical signs of CCPP. The clinical signs of diseased cases revealed serous to mucoid nasal discharge, coughing, dyspnea, frothy salivation, and fever (40-42°C). Necropsied animals showed fibrinous pleuropneumonia and increased pleural fluid. Of 400 nasal swabs, 190 pleural fluid, and 110 lung samples, 26 (6.5%), 31 (16.3%) and 19 (17.3%) Mycoplasma isolates were recovered, respectively. Biochemically, all isolates were sensitive to digitonin and fermented glucose. Sixty seven of Mycoplasma isolates were belonged to Mycoplasma mycoides cluster based on detection of 16S rRNA. Polymerase chain reaction screening of Mycoplasma isolates using specific primer for M. capricolum subsp. capripneumoniae confirmed 55 isolates to be M. capricolum subsp. capripneumoniae., (Copyright © 2017 Elsevier B.V. All rights reserved.)

We have developed teabags packed with dehydrated plant powders, without any supplements, for preparation of plant infusions necessary to develop media for culturing rhizobacteria. These bacteria are efficiently cultivated on such plant teabag culture media, with better progressive in situ recoverability compared to standard chemically synthetic culture media. Combining various plant-based culture media and incubation conditions enabled us to resolve unique denaturing gradient gel electrophoresis (DGGE) bands that were not resolved by tested standard culture media. Based on polymerase chain reaction PCR-DGGE of 16S rDNA fingerprints and sequencing, the plant teabag culture media supported higher diversity and significant increases in the richness of endo-rhizobacteria, namely Gammaproteobacteria (Enterobacteriaceae) and predominantly Alphaproteobacteria (Rhizobiaceae). This culminated in greater retrieval of the rhizobacteria taxa associated with the plant roots. We conclude that the plant teabag culture medium by itself, without any nutritional supplements, is sufficient and efficient for recovering and mirroring the complex and diverse communities of rhizobacteria. Our message to fellow microbial ecologists is: simply dehydrate your plant canopy, teabag it and soak it to prepare your culture media, with no need for any additional supplementary nutrients., (© 2016 Scandinavian Plant Physiology Society.)

Our previous publications and the data presented here provide evidences on the ability of plant-based culture media to optimize the cultivability of rhizobacteria and to support their recovery from plant-soil environments. Compared to the tested chemically-synthetic culture media (e.g. nutrient agar and N-deficient combined-carbon sources media), slurry homogenates, crude saps, juices and powders of cactus (Opuntia ficus-indica) and succulent plants (Aloe vera and Aloe arborescens) were rich enough to support growth of rhizobacteria. Representative isolates of Enterobacter spp., Klebsiella spp., Bacillus spp. and Azospirillum spp. exhibited good growth on agar plates of such plant-based culture media. Cell growth and biomass production in liquid batch cultures were comparable to those reported with the synthetic culture media. In addition, the tested plant-based culture media efficiently recovered populations of rhizobacteria associated to plant roots. Culturable populations of >10(6)-10(8) cfu g(-1) were recovered from the ecto- and endo-rhizospheres of tested host plants. More than 100 endophytic culture-dependent isolates were secured and subjected to morphophysiological identification. Factor and cluster analyses indicated the unique community structure, on species, genera, class and phyla levels, of the culturable population recovered with plant-based culture media, being distinct from that obtained with the chemically-synthetic culture media. Proteobacteria were the dominant (78.8%) on plant-based agar culture medium compared to only 31% on nutrient agar, while Firmicutes prevailed on nutrient agar (69%) compared to the plant-based agar culture media (18.2%). Bacteroidetes, represented by Chryseobacterium indologenes, was only reported (3%) among the culturable rhizobacteria community of the plant-based agar culture medium.

P-Nitrophenol is a major metabolite of some organophosphorus compounds. It is considered to be one of nitrophenol derivatives of diesel exhaust particles that induce substantial hazards impacts on human and animal health. P-Nitrophenol (PNP) is a persistent organic pollutant. Consequently, bioaccumulation of PNP potentiates toxicity. The objectives of the current study were to assess the potential hepatic toxicity and pathway associated with long-term exposure to PNP. Japanese quails were orally administered different doses of PNP for 75 days. Liver and plasma samples were collected at days 45 (45D), days 60 (60D) and days 75 (75D). Liver histological changes and plasma corticosterone levels were assessed. Basal mRNA level of cytochromes P450 (CYP 450) (CYP1A4, 1A5, 1B1), heme oxygenase (HO-1), and aryl hydrocarbon receptor 1 (AhR1), from the liver of exposed birds and primary hepatocytes cultured for 24 hr with PNP, were analyzed using quantitative real-time PCR. The results revealed various histopathological changes in the liver, such as lymphocytes aggregation and hepatocytes degeneration. Significant increases in corticosterone levels were reported. After 60-days of in vivo exposure, the birds exhibited an overexpression in the liver CYP1A4, 1B1, AhR1, and HO-1. Furthermore, with continuous PNP administration, an overall downregulation of the tested genes was observed. In vitro, although a significant overexpression of CYP1A4, 1B1, and HO-1 was observed, CYP1A5 was downregulated. In conclusion, PNP can interfere with the liver CYP 450 enzymes and modulate HO-1 expression in the in vitro and in vivo experiments. Hence, it could have serious deleterious effects on humans, livestock, and wild animals.

P-Nitrophenol (PNP) is considered to be one of nitrophenol derivatives of diesel exhaust particles. PNP is a major metabolite of some organophosphorus compounds. PNP is a persistent organic pollutant as well as one of endocrine-disrupting compounds. Consequently, bioaccumulation of PNP potentiates toxicity. The objectives of the current study were to assess in vivo adverse effects of long-term low doses of PNP exposure on reproductive system during development stage. Twenty-eight-day-old male Japanese quails were orally administered different doses of PNP (0, 0.01, 0.1, 1 mg/kg body weight) daily for 2.5 months. Testicular histopathology, hormones, caspase-3 (CASP3), and claudin-1 (CLDN1) tight junction protein, as well as plasma hormones were analyzed. The results revealed that long-term PNP exposure caused testicular histopathological changes such as vacuolation of spermatogenic cell and spermatocyte with significant testicular and cloacal gland atrophy. PNP activated CASP3 enzyme that is an apoptosis-related cysteine peptidase. Besides, it disrupted the expression of CLDN1. Furthermore, a substantial decrease in plasma concentrations of luteinizing hormone (LH) and testosterone was observed after 2 and 2.5 months in the PNP-treated groups. Meanwhile, the pituitary LH did not significantly change. Site of action of PNP may be peripheral on testicular development and/or centrally on the hypothalamic-pituitary-gonadal axis through reduction of pulsatile secretion of gonadotrophin-releasing hormone. Consequently, it may reduce the sensitivity of the anterior pituitary gland to secrete LH. In conclusion, PNP induced profound endocrine disruption in the form of hormonal imbalance, induction of CASP3, and disruption of CLDN1 expression in the testis. Hence, it may hinder the reproductive processes.

Tilmicosin (TIL) is a long-acting macrolide antibiotic approved for the treatment of cattle with Bovine Respiratory Disease. However, overdose of TIL has been reported to induce cardiotoxicity. The purpose of our experiment was to evaluate the protective effects of Commiphora molmol (mirazid (MRZ); myrrh) and (or) ascorbic acid (AA) against TIL-induced cardiotoxicity in mice. MRZ and AA were orally administered using stomach gavage, either alone or in combination for 5 consecutive days, followed with a single TIL overdose. TIL overdose induced a significant increase in serum levels of cardiac damage biomarkers (AST, LDH, CK, CK-MB, and cTnT), as well as cardiac lipid peroxidation, but cardiac levels of antioxidant biomarkers (GSH, SOD, CAT, and TAC) were decreased. Both MRZ and AA tended to normalize the elevated serum levels of cardiac injury biomarkers. Furthermore, MRZ and AA reduced TIL-induced lipid peroxidation and oxidative stress parameters. MRZ and AA combined produced a synergistic cardioprotective effect. We conclude that myrrh and (or) vitamin C administration minimizes the toxic effects of TIL through their free-radical-scavenging and potent antioxidant activities.

In this study, a potential radiation hazard from TENORM sludge wastes generated during exploration and extraction processes of oil and gas was evaluated. The activity concentration of natural radionuclides (238)U, (226)Ra and (232)Th were determined in TENORM sludge waste. It was found that sludge waste from oil and gas industry is one of the major sources of (226)Ra in the environment. Therefore, some preliminary chemical treatment of sludge waste using Triton X-100 was also investigated to reduce the radioactivity content as well as the risk of radiation hazard from TENORM wastes. The activity concentrations of (226)Ra and (228)Ra in petroleum sludge materials before and after chemical treatment were measured using gamma-ray spectrometry. The average values of the activity concentrations of (226)Ra and (228)Ra measured in the original samples were found as 8908 Bq kg(-1) and 933 Bq kg(-1), respectively. After chemical treatment of TENORM samples, the average values of the activity concentrations of (226)Ra and (228)Ra measured in the samples were found as 7835 Bq kg(-1) and 574 Bq kg(-1), respectively. Activity concentration index, internal index, absorbed gamma dose rate and the corresponding effective dose rate were estimated for untreated and treated samples., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Background: Data are scarce on the advantage of ultrasound-guided spinal anaesthesia in patients with easily identifiable bony landmarks. In this study, we compared the use of ultrasound to the landmark method in patients with no anticipated technical difficulty, presenting for caesarean delivery under spinal anaesthesia., Methods: A total of 150 pregnant women were recruited in this randomized, controlled study. Ultrasound examination and spinal anaesthesia were performed by three anaesthetists with experience in ultrasound-guided neuraxial block. Patients were randomized to either the Ultrasound Group (n=75) or the Landmark Group (n=75). In both groups the level of L3-4 or L4-5 was identified by ultrasound (transverse and longitudinal approach) or palpation. The primary outcome was the procedure time, measured from the time of skin puncture by the introducer to the time of viewing cerebrospinal fluid at the hub of the spinal needle. Secondary outcomes were the number of skin punctures, number of passes, and incidence of successful spinal blockade., Results: The average procedure time, number of skin punctures and needle passes, were similar in both groups. The number of patients with successful spinal anaesthesia after one puncture was not statistically different between the groups., Conclusion: The present results indicate that when performed by anaesthetists experienced in both ultrasound and landmark techniques, the use of ultrasound does not appear to increase the success rate of spinal anaesthesia, or reduce the procedure time or number of attempts in obstetric patients with easily palpable spines., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Organic agriculture as well as good agricultural practices (GAPs) intrigues the concern of both consumers and producers of agricultural commodities. Bio-preparates of various rhizospheric microorganisms (RMOs) are potential sources of biological inputs supporting plant nutrition and health. The response of open-field potatoes to the application of RMO bio-preparates, the biofertilizer "Biofertile" and the bioagent "Biocontrol", were experimented over 5 successive years under N-hunger of north Sinai desert soils. Both vegetative and tuber yields of a number of tested cultivars were significantly improved due to rhizobacterial treatments. In the majority of cases, the biofertilizer "Biofertile" did successfully supply ca. 50% of plant N requirements, as the yield of full N-fertilized plants was comparable to those received 50% N simultaneously with bio-preparates treatment. The magnitude of inoculation was cultivar-dependent; cvs. Valor and Oceania were among the most responsive ones. Bio-preparate introduction to the plant-soil system was successful via soaking of tubers and/or spraying the plant canopy. The "Biocontrol" formulation was supportive in controlling plant pathogens and significantly increased the fruit yields. The cumulative effect of both bio-preparates resulted in tuber yield increases of ca. 25% over control.

North Sinai deserts were surveyed for the predominant plant cover and for the culturable bacteria nesting their roots and shoots. Among 43 plant species reported, 13 are perennial (e.g. Fagonia spp., Pancratium spp.) and 30 annuals (e.g. Bromus spp., Erodium spp.). Eleven species possessed rhizo-sheath, e.g. Cyperus capitatus, Panicum turgidum and Trisetaria koelerioides. Microbiological analyses demonstrated: the great diversity and richness of associated culturable bacteria, in particular nitrogen-fixing bacteria (diazotrophs); the majority of bacterial residents were of true and/or putative diazotrophic nature; the bacterial populations followed an increasing density gradient towards the root surfaces; sizeable populations were able to reside inside the root (endorhizosphere) and shoot (endophyllosphere) tissues. Three hundred bacterial isolates were secured from studied spheres. The majority of nitrogen-fixing bacilli isolates belonged to Bacillus megaterium, Bacillus pumilus, Bacillus polymexa, Bacillus macerans, Bacillus circulans and Bacillus licheniformis. The family Enterobacteriaceae represented by Enterobacter agglomerans, Enterobacter sackazakii, Enterobacter cloacae, Serratia adorifera, Serratia liquefaciens and Klebsiella oxytoca. The non-Enterobacteriaceae population was rich in Pantoae spp., Agrobacterium rdiobacter, Pseudomonas vesicularis, Pseudomonas putida, Stenotrophomonas maltophilia, Ochrobactrum anthropi, Sphingomonas paucimobilis and Chrysemonas luteola. Gluconacetobacter diazotrophicus were reported inside root and shoot tissues of a number of tested plants. The dense bacterial populations reported speak well to the very possible significant role played by the endophytic bacterial populations in the survival, in respect of nutrition and health, of existing plants. Such groups of diazotrophs are good candidates, as bio-preparates, to support the growth of future field crops grown in deserts of north Sinai and irrigated by the water of El-Salam canal.

Among 42 plant species representing the flora of north Sinai, two possessed sand grain sheath encasing the roots. They are Panicum turgidum Forssk. and Stipagrostis scoparia (Trin.and Rupr.) deWinter. Rhizosheaths, compared to surrounding free sand, accommodated higher population density of microorganisms including associative diazotrophs. Isolates secured belonged to the species of Bacillus circulans, Paenib. macerans (Bacillus macerans), Enterobacter agglomerans, Agrobacterium radiobacter and Chryseomonas luteola (Pseudomonos luteola). The rhizosheath potentiality in re-vegetating sand dunes and arid lands, through nitrogen fixation, plant-water relationship and root continuity for nutrient uptake, are discussed.

Studies were conducted into the effects on pituitary gonadotrophic hormones in ovariectomized rats of butorphanol, a synthetic morphine derivative which was claimed to be a potent analgesic with few side-effects, in comparison to effects of the naturally occurring alkaloid morphine. For this purpose, 3 groups of ovariectomized rats were used. Rats of the 1st group were injected butorphanol at 2 dose levels (1 or 2 mg/kg body weight [b.w.]. Those of the 2nd group were injected morphine sulphate (10 or 20 mg/kg b.w.). The 3rd group was injected saline and served as control. Blood samples were collected by orbital sinus punctures, just before treatment and 1 hour post injection. Luteinizing hormone (LH) and follicle-stimulation hormone levels were determined in the sera of rats by radio-immuno-assay. The results revealed that morphine, at the 2 dose levels used, produced more than 90% decrease in serum LH concentration, whereas butorphanol produced more than 70% decrease in serum LH levels. Both morphine and butorphanol, at the 2 doses used, produced more than 76% decrease in serum follicle stimulating hormone concentration. It is concluded that butorphanol, the morphinic derivative, has a depressive effect on the synthesis and/or release of gonadotrophic hormones. This inhibitory effect, however, was nearly as potent as that produced by morphine sulphate.

The validity of seed bacterization with Azotobacter chroococcum in soils of variable densities of naturally present azotobacters was studied. Inoculation of barley grains had no effect on counts of total microflora, neither in rhizosphere nor in root-free soil, but significantly increased Azotobacter population, especially in the rhizosphere. The rate of colonization in the root region was much higher when soil initially harboured low Azotobacter densities. Bacterization improved plant growth and increased soil nitrogen through nitrogen fixation. Nitrogen balance in soils showed higher gains in the inoculated treatments over the uninoculated analogues of 30--98 ppm.