Outcomes of dietary alpha-lipoic acid on testicular vascularization, steroid hormones, and seminal quality in aged Baladi bucks.

BACKGROUND: Senescence is accompanied by a progressive decrease in male reproductive performance, mainly due to oxidative stress and endothelial dysfunction. Alpha lipoic acid (ALA) is a potent antioxidant, that diffuses freely in aqueous and lipid phases, possessing anti-inflammatory and anti-apoptotic properties. This study aimed to examine the effects of supplemental dietary ALA on testicular hemodynamics (TH), circulating hormones, and semen quality in aged goats. Twelve Baladi bucks were divided into two groups (n = 6 each); the first fed a basic ration and served as a control group (CON), while the second received the basic ration supplemented with 600 mg ALA/ kg daily for consecutive eight weeks (ALA). RESULTS: There were improvements in testicular blood flow in the ALA group evidenced by a lower resistance index (RI) and pulsatility index (PI) concurrent with higher pampiniform-colored areas/pixel (W3-W6). There were increases in testicular volume and decreases in echogenicity (W3-W5; ALA vs. CON). Compared to the CON, ALA-bucks had higher serum concentrations of testosterone, estradiol, and nitric oxide (W3-W5). There were enhancements in semen traits (progressive motility, viability, morphology, and concentration, alanine aminotransferase enzyme) and oxidative biomarkers (catalase, total antioxidant capacity, and malondialdehyde). CONCLUSIONS: ALA dietary supplementation (600 mg/kg diet) improved aged bucks' reproductive performance by enhancing the testicular volume, testicular hemodynamics, sex steroids, and semen quality.

A pilot study on ultrashort peptide with fluconazole: A promising novel anticandidal combination.

Background and Aim: Human infections caused by Candida albicans are common and range in severity from relatively treatable skin and mucosal conditions to systemic, fatal invasive candidiasis. The treatment of fungal infections is challenged by major obstacles, including the scarcity of effective therapeutic options, the toxicity of available medications, and the escalating antifungal resistance. Hence, there exists an urgent need to develop new classes of antimicrobial agents. This study was conducted to investigate the effect of KW-23 peptide against standard and resistant strains of C. albicans alone and in combination with fluconazole. Materials and Methods: A conjugated ultrashort antimicrobial peptide (KW-23) was designed and synthesized. KW-23 was challenged against standard and multidrug-resistant C. albicans alone and in combination with fluconazole using standard antimicrobial and checkerboard assays. The toxicity of the peptide was examined using hemolytic assays. Results: KW-23 positively affected the standard and resistant Candidal strains (at 5 and 15 µg/mL respectively), exhibiting potent synergistic antimicrobial activity against the standard strain when combined with fluconazole. The effect of the combination was additive against the resistant strain (0.6 µg/mL). Furthermore, the peptide exhibited negligible toxicity on human erythrocytes. Conclusion: KW-23 and its combination with fluconazole could be a promising candidate for developing anticandidal agents.

Influence of the Addition of Zinc Oxide Nanoparticles to Cryopreservation Medium for Dog Epididymal Spermatozoa.

The purpose of this study was to assess the effect of different concentrations of zinc oxide nanoparticles (ZnONPs) introduced to an extender for frozen-thawed epididymal dog spermatozoa. Epididymides from 22 castrated dogs were minced and cultured in a Tris buffer. The recovered spermatozoa were diluted in Tris-Citric acid-Fructose (TCF) extender with different concentrations of ZnONPs (100 and 200 µg/mL) and control (0.0 µg/mL). Diluted samples were equilibrated at 5 °C for 2 hours before being packed in 0.25 mL straws and stored in liquid nitrogen (-196 °C). After thawing at 37°C for 30 seconds, sperm motility, viability, membrane integrity, acrosome integrity, DNA integrity, and lipid peroxidation by malondialdehyde (MDA) concentration were all measured. The results were presented as mean ± SEM. Adding 100 and 200 µg/mL ZnONPs to the cryopreservation medium significantly (P < .05) improved motility and membrane integrity compared to the control. Viability and acrosome integrity were considerably (P < .05) better at 100 µg/mL ZnONPs than at 200 µg/mL ZnONPs and the control. MDA concentration was significantly (P < .05) decreased at 100 µg/mL ZnONPs compared to 200 µg/mL ZnONPs and the control. When 100 µg/mL ZnONPs were compared to 200 µg/mL ZnONPs and the control, the percentage of DNA damage was significantly (P < .05) reduced. Consequently, adding 100 µg/mL ZnONPs to TCF extender resulted in a significant increase in the proportion of motility, viability, membrane-intact, and acrosome-intact dog epididymal sperm, as well as the preservation of DNA integrity and the prevention of lipid peroxidation at the membrane level.

Single melatonin injection enhances the testicular artery hemodynamic, reproductive hormones, and semen parameters in German shepherd dogs.

This study aimed to determine the effects of melatonin administration on testicular vascular perfusion in relation to steroid hormones and semen characteristics in dogs. The study included 12 normospermic German shepherd dogs (weighed 35 ± 0.5 kg and aged 4 ± 0.5 years). Males received a single melatonin administration (melatonin dimethyl sulfoxide + corn oil via subcutaneous route; MEL; n = 6), while the rest of the animals served as controls (dimethyl sulfoxide + corn oil; Control; n = 6). Males were subjected to routine examination on days -15, 0, 15, 30, 45, and 60. All examined dogs were subjected to Doppler screening, semen collection, and blood sampling. The MEL group showed a significant (P < 0.05) elevation in semen volume, concentration, percentage of sperm motility, and total sperm × 106 / ejaculate compared to other control males. Doppler indices as resistance (RI) and pulsatility (PI) indices declined (P < 0.05) from D 30 (1.02 ± 0.01) until day 60 (0.87 ± 0.02) of treatment. In MEL males, the peak systolic point of velocity (PSV; cm/sec) of the testicular artery elevated (P < 0.05) on day 60 (20.15 ± 0.99) compared to its value on day 0 (17.39 ± 1.84). On D 60, the levels of testosterone (T), estradiol 17-ß (E2), and nitric oxide (NO) elevated (P < 0.05). A negative correlation was detected between testicular volume, scrotal circumference (SC), T levels, Doppler indices, and velocities. In conclusion, single melatonin administration could improve testicular vascularization via increasing Doppler velocities and intratesticular colored areas. In addition, it could improve semen picture and steroids (T and E2) and nitric oxide.

Study the Effect of Conjugate Novel Ultra-Short Antimicrobial Peptide with Silver Nanoparticles against Methicillin Resistant S. aureus and ESBL E. coli.

Background: Bacterial resistance is a challenging limitation in infection treatment. This work evaluates the potential antibacterial activity of conjugation of Tryasine peptide with silver nanoparticles against selected pathogens. Materials and Methods: The peptide Tryasine was produced using three subunits of tryptophan and three lysine amino acids, then its purity was determined by reverse-phase high-performance liquid chromatography. The peptide was confirmed using mass spectrometry and electrospray ionization mass spectrometry. Silver nanoparticles conjugate with Tryasine was synthesized by adding Tryasine-silver nitrate solution in the presence of the reducing agent sodium borohydride. The presence of Tryasine-silver nanoparticles was indicated by the yellow-brown color and was further confirmed through ultraviolet-visible spectrophotometry. The minimum inhibitory and minimum bactericidal concentrations for Tryasine nanoparticles were determined against Staphylococcus aureus, Escherichia coli, methicillin resistant Staphylococcus aureus, and ESBL Escherichia coli using the microdilution method. Toxicity for nanoparticles conjugated with Tryasine was determined using erythrocyte hemolytic assay. Results: Tryasine alone was effective (MIC around 100 and 200 µM) against standard and resistant strains of bacteria used. However, Tryasine-silver nanoparticles were more effective with MICs ranging from 30 to 100 µM depending on the bacterial strain used. Tryasine-silver nanoparticles at concentration of 100 µM only caused 1% hemolysis on human erythrocytes after 30 min of incubation. Conclusions: The findings indicate that Tryasine-silver nanoparticles had good antibacterial activity against pathogenic strains of Gram-positive and Gram-negative bacteria. Additionally, the conjugate showed low hemolytic activity and cytotoxicity. Therefore, conjugation of Tryasine with silver nanoparticles is a promising treatment candidate for bacterial infection with low toxicity.

Knowledge, Attitudes, and Practices of Adult Iraqi Population Towards COVID-19 Booster Dose: A Cross-Sectional Study.

Purpose: COVID-19 vaccines are critical for containing the pandemic and preventing serious SARS-CoV-2 infections. In addition to the two main doses, a booster dose has been utilized to improve immunity. The aim of current study is to evaluate Iraqi adult population knowledge, attitudes, and practices towards COVID-19 booster dose. Subjects and Methods: This online cross-sectional survey of adult Iraqis (n = 754) assessed the attitudes of people who have had both immunizations regarding a potential COVID-19 vaccine booster dosage and to identify potential factors that might impact these attitudes. Factors evaluated in the current study included previously received vaccine type in the first two doses, socioeconomic characteristics, health status, knowledge about COVID-19 and its vaccines and adherence to protective practices. Results: Overall, 61.1% of participants expressed willingness to receive a COVID-19 booster dose, with a high median score of knowledge and practice toward COVID-19. Participants who did not perceive COVID-19 to be serious, p-value <0.001), participants who believed they would not be infected with COVID-19 in the next 6 months (p-value <0.001), low knowledge score group (p-value <0.001), lower education (p-value <0.001), participants who received the COVID-19 vaccine because of imposed laws (p-value <0.001), participants who received AstraZeneca vaccine (p-value <0.001), younger participants (p-value=0.003), low level of practice (p-value <0.001), participants who did not know someone who had died due to COVID-19 (p-value=0.01), low risk of developing serious side effects if infected with COVID-19 and participants in the low side effects score were significantly less frequently willing to receive a booster COVID-19 dose (p-value <0.001). The main reasons for booster dose hesitancy/refusal were the perceived lack of need for a booster shot, the uselessness of a booster shot and the conspiracy theory of boosting corporate profits through booster shots. Conclusion: There is high hesitancy towards COVID-19 booster dose acceptance among the Iraqi population. The study identified several factors associated with vaccine hesitancy including low socioeconomic status and low knowledge about COVID-19 and its vaccines.

The efficiency of intrauterine infusion of platelet-rich plasma in the treatment of acute endometritis as assessed by endoscopic, Doppler, oxidative, immunohistochemical, and gene expression alterations in jennies.

This study used autologous platelet-rich plasma (PRP) to treat acute endometritis in jennies with follow-up for alterations in uterine hemodynamics, endoscopic, immunohistochemistry, oxidant/antioxidant imbalance, pro-inflammatory regulatory molecules, and transmembrane mucin expressions. Ten jennies suffering from endometritis (acute type; n = 10) were included in the study. PRP was prepared from each animal and two intrauterine infusions one week apart were administrated. Examination and follow-up were done physically, ultrasonographically, endoscopically and samples were taken for histopathology, immunohistochemistry, and bacteriological examination. Blood and uterine fluid samples were taken to estimate biochemical and oxidative stress alterations. Expression of TRAF6 and MUC1 genes was investigated in uterine fluid, at days -1 (day of diagnosis establishment), 7, 14, and 21. Uterine bacteriological examination showed a decrease in bacterial isolates after PRP treatment. The uterine thickness and uterine vascular perfusion as illustrated by color Doppler ultrasonography were significantly decreased in jennies treated by PRP. Uterine spectral wave pattern showed a significant linear increase in pulsatility index only. Three weeks after first PRP treatment, white light endoscopic examination revealed normal uterine body mucosa and uterine horn folds. A high nuclear factor (NF-κB) expression was seen in the mononuclear cells. A significant reduction in oxidative stress biomarkers in both serum and uterine fluid was recorded after PRP treatment. The TRAF-1 gene expression significantly decreased gradually after intrauterine PRP infusion. The MUC-1 gene expression significantly decreased gradually after intrauterine PRP infusion. Both genes were within normal levels by week 3. Endometritis in jennies is associated with an oxidative process, alterations in serum biochemical parameters, Doppler indices, endoscopic appearance, high NF-κB expression, and upregulation of TRAF-1 and MUC-1 expressions. Two intrauterine infusions of autologous PRP restored normal endometrial appearance after acute endometritis.

The design and evaluation of the antimicrobial activity of a novel conjugated penta-ultrashort antimicrobial peptide in combination with conventional antibiotics against sensitive and resistant strains of S. aureus and E. coli.

Background and purpose: Antimicrobial resistance still constitutes a major health concern to the global human population. The development of new classes of antimicrobial agents is urgently needed to thwart the continuous emergence of highly resistant microbial pathogens. Experimental approach: In this study, we have rationally designed a novel conjugated ultrashort antimicrobial peptide. The peptide named naprolyginine was challenged against representative strains of wild-type and multidrug-resistant bacteria individually or in combination with individual antibiotics by employing standard antimicrobial and checkerboard assays. Findings / Results: Our results displayed that the peptide exhibits potent synergistic antimicrobial activity against resistant strains of gram-positive and gram-negative bacteria when combined with individual antibiotics. Additionally, the peptide was evaluated for its hemolytic activity against human red blood cells and displayed negligible toxicity. Conclusion and implications: Naprolyginine could prove to be a promising candidate for antimicrobial drug development.

Selenium nanoparticles from Lactobacillus paracasei HM1 capable of antagonizing animal pathogenic fungi as a new source from human breast milk.

The current study was performed to develop a simple, safe, and cost-effective technique for the biosynthesis of selenium nanoparticles (SeNPs) from lactic acid bacteria (LAB) isolated from human breast milk with antifungal activity against animal pathogenic fungi. The LAB was selected based on their speed of transforming sodium selenite (Na2SeO3) to SeNPs. Out of the four identified LAB isolates, only one strain produced dark red color within 32 h of incubation, indicating that this isolate was the fastest in transforming Na2SeO3 to SeNPs; and was chosen for the biosynthesis of LAB-SeNPs. The superior isolate was further identified as Lactobacillus paracasei HM1 (MW390875) based on matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and phylogenetic tree analysis of 16S rRNA sequence alignments. The optimum experimental conditions for the biosynthesis of SeNPs by L. paracasei HM1 were found to be pH (6.0), temperature (35˚C), Na2SeO3 (4.0 mM), reaction time (32 h), and agitation speed (160 rpm). The ultraviolet absorbance of L. paracasei-SeNPs was detected at 300 nm, and the transmission electron microscopy (TEM) captured a diameter range between 3.0 and 50.0 nm. The energy-dispersive X-ray spectroscopy (EDX) and the Fourier-transform infrared spectroscopy (FTIR) provided a clear image of the active groups associated with the stability of L. paracasei-SeNPs. The size of L. paracasei-SeNPs using dynamic light scattering technique was 56.91 ± 1.8 nm, and zeta potential value was -20.1 ± 0.6 mV in one peak. The data also revealed that L. paracasei-SeNPs effectively inhibited the growth of Candida and Fusarium species, and this was further confirmed by scanning electron microscopy (SEM). The current study concluded that the SeNPs obtained from L. paracasei HM1 could be used to prepare biological antifungal formulations effective against major animal pathogenic fungi. The antifungal activity of the biologically synthesized SeNPs using L. paracasei HM1 outperforms the chemically produced SeNPs. In vivo studies showing the antagonistic effect of SeNPs on pathogenic fungi are underway to demonstrate the potential of a therapeutic agent to treat animals against major infectious fungal diseases.

Genetic and antimicrobial resistance profiles of non-O157 Shiga toxin-producing Escherichia coli from different sources in Egypt.

BACKGROUND: The Shiga toxin-producing Escherichia coli (STEC) represented a great risk to public health. In this study, 60 STEC strains recovered from broiler and duck fecal samples, cow's milk, cattle beef, human urine, and ear discharge were screened for 12 virulence genes, phenotypic and genotypic antimicrobial resistance, and multiple-locus variable-number tandem-repeat analysis (MLVA). RESULTS: The majority of strains harbored Shiga toxin 1 (stx1) and stx1d, stx2 and stx2e, and ehxA genes, while a minority harbored stx2c subtype and eaeA. We identified 10 stx gene combinations; most of strains 31/60 (51.7%) exhibited four copies of stx genes, namely the stx1, stx1d, stx2, and stx2e, and the strains exhibited a high range of multiple antimicrobial resistance indices. The resistance genes blaCTX-M-1 and blaTEM were detected. For the oxytetracycline resistance genes, most of strains contained tetA, tetB, tetE, and tetG while the tetC was present at low frequency. MLVA genotyping resolved 26 unique genotypes; genotype 21 was highly prevalent. The six highly discriminatory loci DI = 0.9138 are suitable for the preliminary genotyping of STEC from animals and humans. CONCLUSIONS: The STEC isolated from animals are virulent, resistant to antimicrobials, and genetically diverse, thus demands greater attention for the potential risk to human.

The Design of Alapropoginine, a Novel Conjugated Ultrashort Antimicrobial Peptide with Potent Synergistic Antimicrobial Activity in Combination with Conventional Antibiotics.

(1) Background: Antimicrobial resistance represents an urgent health dilemma facing the global human population. The development of novel antimicrobial agents is needed to face the rising number of resistant bacteria. Ultrashort antimicrobial peptides (USAMPs) are considered promising antimicrobial agents that meet the required criteria of novel antimicrobial drug development. (2) Methods: Alapropoginine was rationally designed by incorporating arginine (R), biphenylalanine (B), and naproxen to create an ultrashort hexapeptide. The antimicrobial activity of alapropoginine was evaluated against different strains of bacteria. The hemolytic activity of alapropoginine was also investigated against human erythrocytes. Finally, synergistic studies with antibiotics were performed using the checkerboard technique and the determination of the fractional inhibitory index. (3) Results: Alapropoginine displayed potent antimicrobial activities against reference and multi-drug-resistant bacteria with MIC values of as low as 28.6 µg/mL against methicillin-resistant S. aureus. Alapropoginine caused negligible toxicity toward human red blood cells. Moreover, the synergistic studies showed improved activities for the combined conventional antibiotics with a huge reduction in their antimicrobial concentrations. (4) Conclusions: The present study indicates that alapropoginine exhibits promising antimicrobial activity against reference and resistant strains of bacteria with negligible hemolytic activity. Additionally, the peptide displays synergistic or additive effects when combined with several antibiotics.

Effect of melatonin supplementation during IVM of dromedary camel oocytes (Camelus dromedarius) on their maturation, fertilization, and developmental rates in vitro.

The positive impact of melatonin on in vitro embryo production (IVEP) has been reported in many domestic species; however, no studies have been carried out in camelids. We aimed to evaluate the effects of melatonin supplementation in maturation media on in vitro maturation, fertilization, and preimplantation embryo development of dromedary camel oocytes (experiment 1). We also evaluated the concentrations of total antioxidant capacity (TAC), and malondialdehyde (MDA) in the IVM spent medium in relation to melatonin supplementation. Cumulus oocyte complexes (COCs) were cultured in in vitro maturation media (IVM) supplemented with either 0.0, 25.0, 50.0 or 75.0 µM of melatonin for 30 h. Matured oocytes were then fertilized in vitro with epididymal camel spermatozoa. Following IVF, the resulting embryos were cultured in vitro for seven days. The percentage of maturation, fertilization, cleavage, and embryo developmental rates (morula and blastocyst) was recorded (experiment 1). TAC and MDA levels in the IVM spent maturation media were also evaluated at 30 h post-IVM (experiment 2). The results showed that supplementation of IVM media with 25 µM melatonin significantly improved oocyte nuclear maturation, fertilization (18 h post-insemination; pi), cleavage (day 3 pi), morula (day 5 pi) and blastocyst (day 7 pi) rates as compared with the controls and other melatonin-supplemented groups. Furthermore, the TAC in the IVM spent media was significantly increased (P < 0.05) in 25 µM melatonin supplemented groups than those supplemented with 0.0, 50.0, 75.0 µM melatonin. However, the concentration of MDA was significantly lower (P < 0.05) in IVM media supplemented with 25.0 µM of melatonin when compared with the control and other treatment groups. In conclusion, supplementation of IVM medium with 25 µM of melatonin could enhance the in vitro developmental capacity of dromedary camel oocytes.

Beneficial Effects of L-Carnitine Supplementation during IVM of Canine Oocytes on Their Nuclear Maturation and Development In Vitro.

This study aimed to investigate the effect of L-Carnitine (LC) supplementation during in vitro maturation (IVM) of canine oocytes on nuclear maturation, fertilization status, and preimplantation development. Cumulus-oocyte complexes (COCs) collected from the ovaries of ovariohysterectomized female dogs were matured in vitro for 72 h in a TCM-199 medium supplemented with (0.1, 0.3, 0.6, 1.0, or 2.0 mg/mL) or without (0.0 mg/mL) LC. Matured oocytes were fertilized in vitro with frozen-thawed spermatozoa, and zygotes were cultured in a SOF medium for 7 days. IVM rates were higher (p ≤ 0.05) in 0.3 and 0.6 mg/mL LC supplemented groups than in the control (0.0 mg/mL LC) and other LC groups. Fertilization (18 h postinsemination (pi)) and cleavage (2-16-cell stage at day 3 pi) rates were higher (p ≤ 0.05) in the 0.6 mg/mL LC group than in the control and 0.1, 1.0, and 2 mg/mL LC supplemented groups. Interestingly, 4.5% of fertilized oocytes developed to morula (day 5 pi) in the 0.6 mg/mL LC group, which was higher (p ≤ 0.05) than those developed in the 0.3 mg/mL group (1.0%). No cleaved embryos developed to morula in other groups. In conclusion, LC supplementation at 0.6 mg/mL during IVM of canine oocytes improved their maturation, fertilization, and preimplantation embryo development rates following IVF and in vitro culture (IVC).

Alleviation of carbon tetrachloride-induced hepatocellular damage and oxidative stress in rats by Anabaena oryzae phycocyanin.

Phycocyanin isolated from Anabaena biomass was in-vitro assayed for its antioxidant activity against DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS [2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)] free radical, revealing maximum activities amounting to 77 and 80% at 1,000 µg/ml and SC50 values about 96 and 111 µg/ml, respectively. A biological experiment was conducted, involving 40 male Wistar Albino rats, divided into five groups. Group I received only the basal diet as a normal control, while groups II, III, IV, and V were administrated intraperitoneal (IP) injection of a single dose of CCl4 (50% in corn oil) at 0.5 ml/kg body weight. Subsequently, groups II, III, IV, and V received phycocyanin at 0.0, 25, 50, and 100 mg/kg body weight/day. CCl4 induced considerable increases (p < .05) in the levels of serum ALT, AST, urea and creatinine, total lipid, and triglycerides coupled with significant reductions (p < .05) in serum antioxidant enzymes and some liver histopathological deformations compared to the negative control (group 1). Administration of Anabaena oryzae phycocyanin can counteract these CCl4 -induced changes. PRACTICAL APPLICATIONS: Phycocyanin isolated from Anabaena has beneficial effects such as the antioxidant, antibacterial, anticancer, and hepatoprotective effect. Phycocyanin may play a key role in alleviating oxidative stress, artificially induced by carbon tetrachloride in Albino rats, to ultimately determine its capacity to serve as a natural antioxidant for food and health applications.

Transfer of Anthocyanin Accumulating Delila and Rosea1 Genes from the Transgenic Tomato Micro-Tom Cultivar to Moneymaker Cultivar by Conventional Breeding.

Delila and Rosea1 anthocyanin accumulation genes were subjected to bioinformatics analysis. Delila protein has 56-69% similarity with different anthocyanin-rich plants, while Rosea1 protein has 83-87% with anthocyanin-rich plant proteins. This study aimed at transferring Delila and Rosea1 genes from the transgenic Micro-tom tomato cultivar to the Moneymaker tomato cultivar using traditional breeding for enhancing their fruit anthocyanin content. Results of all produced F1 plants of manual hybridization between both cultivars were consistent with the Mendelian inheritance hypothesis. Plants of F2 populations showed a 3:1 Mendelian segregation proportion (75% of plants have anthocyanin pigmentation). Seeds of F2 were individually cultured to get four homozygous lines with anthocyanin accumulation in fruits. The total anthocyanin in the anthocyanin-enriched inbred fruit (3 g/kg DM) represented a relative increase of about 131% of the parent level. The total phenolic compounds in inbred tomato fruits were 54.9 mg/100 g DM referring to a relative increase of about 51% of the respective parent plant. The antioxidant activity of inbred fruit at maturity (m) was 83.5% compared with 91% for TBHQ. The inbred (m) tomato fruit extract reduced the growth of G- bacteria G+ bacteria by 99% and 95%, respectively.

Simultaneous H2S mitigation and methanization enhancement of chicken manure through the introduction of the micro-aeration approach.

The impact on H2S alleviation and methane yield enhancement after submitting the anaerobic digestion of chicken manure to a finite amount of air was investigated. The largest reduction in the H2S biogas content (58% lower) occurred when air intensity of 30 ml/g VSin was injected into the reactors. Consequently, a maximum methane yield (335 mL-g VSin-1), which was 77% higher than the control, was concurrently achieved. Slight sulfate accumulation (<330 mg L-1) was observed inside the micro-aerated digesters with higher air intensities, suggesting a suppression of sulfide inhibition. Bacterial diversity/richness was enhanced in these digesters while the relative abundance of Methanocelleus increased by 36%. The most important contributing factor to enhancement was the synergistic effect resulting from increments in the hydrolysis rate and the suppression of sulfide inhibition. The results highlighted the potential of in situ H2S mitigation with the added benefit of methane yield enhancement.

New Approach for Controlling Snail Host of Schistosoma mansoni, Biomphalaria alexandrina with Cyanobacterial Strains-Derived C-Phycocyanin.

Schistosomiasis is one of the major communicable diseases of public health and socioeconomic importance in the developing world. It is a waterborne disease in which Biomphalaria alexandrina snails are known to be the intermediate molluscan host for Schistosoma mansoni: the causative agent of human intestinal schistosomiasis. Therefore, snail control is one of the cornerstones of schistosomiasis control programs. Several methods have been used to eliminate snail hosts. One of these methods is chemical molluscicides, which have undesirable effect to nontarget organisms. Consequently, the search for biologically derived molluscicides to complement the use of synthetic molluscicides is a top priority. In this concern, this study is the first to evaluate the molluscicidal potency of Cyanobacterial Phycocyanin (C-PC) as a virtually untapped source. Laboratory assessment of three freshwater Cyanobacterial strains: Anabaena oryzae SOS13, Nostoc muscorum SOS14, and Spirulina platensis SOS13-derived C-Phycocyanin as a biocontrol agent against freshwater mollusks; B. alexandrina snails were performed. Also, the safety of tested C-PC on nontarget organisms (Tilapia fish) was assessed. Results reveal that C-PC extracted from all tested Cyanobacteria strains showed a promising molluscicidal activity (the mortality rate was 100% at 100 µg/mL concentration). Out of the examined strains, A. oryzae SOS13 phycocyanin was found to be the most potent strain (LC50 and LC90 were 38.492 and 49.976 µg/mL, respectively). Moreover, C-PC extracts from all tested strains have been found to be safe to Tilapia fish as the survival rate was 100% at the effective molluscicidal concentrations. We can conclude that C-PC extracts are the first promising microbial biopesticides for the control of freshwater B. alexandrina snails.

Developmental competence of Dromedary camel (Camelus dromedarius) oocytes selected using brilliant cresyl blue staining.

The objectives of the present studies were to investigate the developmental capacity of dromedary camel oocytes selected by brilliant cresyl blue (BCB) staining and to investigate the expression of select transcripts in germinal vesicle (GV) stage oocytes. These transcripts included BMP15 and GDF9 as important transcripts for folliculogenesis and oocyte development, Zar1 and Mater as maternal transcripts required for embryonic development, Cyclin B1 and CDK1 as cell cycle regulators and Oct4 and STAT3 as transcription factors. Dromedary camel oocytes were retrieved from ovaries collected at a local slaughterhouse. After exposure to BCB staining, cumulus-oocyte complexes (COCs) from BCB+, BCB- and control (selected based on morphological criteria) groups were subjected to in vitro maturation, in vitro fertilization and in vitro culture. For gene expression studies, after BCB staining cumulus cells were stripped off and the completely denuded GV stage oocytes were used for RT-PCR analysis of selected transcripts. BCB+ oocytes showed higher maturation, and fertilization rates compared with BCB- and control groups. Indices of early embryonic development, namely, cleavage at 48 hours post insemination (hpi), and development to morula at day 5 and day 7 blastocyst rates were also significantly higher in the BCB+ group. RT-PCR revealed a higher expression of BMP15, GDF9, Zar1, Mater, Cyclin B1, CDK1, OCT4 and STAT3 in good quality oocytes that stained positively for BCB (BCB+). Collectively, results provide novel information about the use of BCB screening for selecting good quality oocytes to improve in vitro embryo production in the dromedary camel.

Pathogenicity of Highly Pathogenic Avian Influenza Virus H5N1 in Naturally Infected Poultry in Egypt.

Highly pathogenic avian influenza virus (HPAIV) H5N1 has been endemic in Egypt since 2006, and there is increasing concern for its potential to become highly transmissible among humans. Infection by HPAIV H5N1 has been described in experimentally challenged birds. However, the pathogenicity of the H5N1 isolated in Egypt has never been reported in naturally infected chickens and ducks. Here we report a 2013 outbreak of HPAIV H5N1 in commercial poultry farms and backyards in Sharkia Province, Egypt. The main symptoms were ecchymosis on the shanks and feet, cyanosis of the comb and wattles, subcutaneous edema of the head and neck for chickens, and nervous signs (torticollis) for ducks. Within 48-72 hrs of the onset of illness, the average mortality rates were 22.8-30% and 28.5-40% in vaccinated chickens and non-vaccinated ducks, respectively. Tissue samples of chickens and ducks were collected for analyses with cross-section immunohistochemistry and real-time RT-PCR for specific viral RNA transcripts. While viral RNA was detected in nearly all tissues and sera collected, viral nucleoprotein was detected almost ubiquitously in all tissues, including testis. Interestingly, viral antigen was also observed in endothelial cells of most organs in chickens, and clearly detected in the trachea and brain in particular. Viral nucleoprotein was also detected in mononuclear cells of various organs, especially pulmonary tissue. We performed phylogenetic analyses and compared the genomic sequences of the hemagglutinin (HA) and nonstructural proteins (NS) among the isolated viruses, the HPAIV circulated in Egypt in the past and currently, and some available vaccine strains. Further analysis of deduced amino acids of both HA and NS1 revealed that our isolates carried molecular determinants of HPAIV, including the multibasic amino acids (PQGERRRK/KR*GLF) in the cleavage site in HA and glutamate at position 92 (D92E) in NS1. This is the first report of the pathogenicity of the HPAIVH5N1 strain currently circulating in naturally infected poultry in Egypt, which may provide unique insights into the viral pathogenesis in HPAIV-infected chickens and ducks.