Isolation, Molecular, and Histopathological Patterns of a Novel Variant of Infectious Bursal Disease Virus in Chicken Flocks in Egypt.

After an extended period of detecting classical virulent, attenuated, and very virulent IBDV, a novel variant (nVarIBDV) was confirmed in Egypt in this study in 18, IBD vaccinated, chicken flocks aged 19-49 days. Partial sequence of viral protein 2 (VP2) [219 aa, 147-366, resembling 657 bp] of two obtained isolates (nos. 3 and 4) revealed nVarIBDV (genotype A2d) and OR682618 and OR682619 GenBank accession numbers were obtained. Phylogenetic analysis revealed that both nVarIBDV isolates were closely related to nVarIBDV strains (A2d) circulating in China, exhibiting 100% identity to SD-2020 and 99.5-98.1% similarity to ZD-2018-1, QZ, GX and SG19 strains, respectively. Similarity to USA variant strains, belonging to genotypes A2b (9109), A2c (GLS) and A2a (variant E), respectively, was 95.5-92.6%. Also, the VP2 hypervariable region in those two, A2d, isolates revealed greater similarities to Faragher 52/70 (Vaxxitek®) at 90.4% and to an Indian strain (Ventri-Plus®) and V217 (Xtreme®) at 89.7% and 86-88.9% in other vaccines. Histopathological examination of both the bursa of Fabricius and spleen collected from diseased chickens in flock no. 18 revealed severe atrophy. In conclusion, further studies are required to investigate the epidemiological situation of this novel genotype across the country, and to assess various vaccine protections against nVarIBDV. Additionally, vaccination of breeders with inactivated IBD vaccines including this nVarIBDV is essential to obtain specific maternal antibodies in their broilers.

Evaluation of faecal lactoferrin as a morbidity biomarker in Schistosoma mansoni infection.

OBJECTIVE: Lactoferrin is an 80 KDa iron-binding glycoprotein that plays a significant role in the innate immune system and is considered to be an important microbicide molecule. This study aimed to assess the concentration of lactoferrin in Schistosoma mansoni-infected cases before and after praziquantel treatment. METHODS: A cross-sectional study was carried out on 250 individuals aged from 5 to 30 years. Stool samples were examined for the presence of parasitic infections using Kato-Katz and formalin ethyl acetate techniques. All S. mansoni-positive cases were treated with praziquantel and stool samples were recollected 21 days later. Faecal lactoferrin level was determined before and after treatment. RESULTS: The prevalence of S. mansoni infection was 14.4%. Among 36 participants infected with S. mansoni, the cure rate was 91.7%. A statistically significant difference in the mean lactoferrin level before and after treatment was detected (1648.95 pg/ml ± 656.5 vs. 1162.8 pg/ml ± 356.8). This difference was statistically significant in the middle and older age groups, in males and in the absence of coinfection with other parasites. CONCLUSION: Lactoferrin could be a promising biomarker associated with S. mansoni infection, however, it could not be used to assess the severity of infection.

Comparison of gdh polymerase chain reaction-restriction fragment length polymorphism and tpi assemblage-specific primers for characterization of Giardia intestinalis in children.

Background: Giardia is a diarrheagenic eukaryotic parasite that consists of at least eight morphologically identical but genetically distinct genotypes. Human giardiasis is caused mainly by A and B assemblages. Aim and objectives: The study aimed to compare the performance of gdh polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and tpi assemblage-specific primers in genotyping of G. intestinalis. Materials and Methods: Stool samples of 315 children were microscopically screened for G. intestinalis. Positive samples were genotyped using tpi assemblage-specific primers and gdh semi-nested PCR-RFLP techniques. Results: The prevalence of Giardia was 18.1%. The detected genotypes using tpi and gdh approaches were assemblage A (15.8% vs. 12.7%) and assemblage B (36.8% vs. 74.5%) as single infections and mixed assemblages A and B (47.4% vs. 12.7%). The two approaches showed a moderate agreement (kappa index = 0.413, P < 0.001). PCR-RFLP of gdh gene revealed that sub-assemblages BIII and BIV were equally detected (30.9% each). The remaining samples were equally divided between sub-assemblage AII, mixed BIII and BIV, and mixed AII and BIII (12.7% each). A significant association was detected between the retrieved sub-assemblages and the presence of symptoms. Conclusions: Although both approaches confirmed the predominance of assemblage B, the use of assemblage-specific primers is more effective in elucidating the true picture of mixed assemblage infection.

Heat stress during critical windows of the oestrous cycle and risk of pregnancy establishment in embryo-recipient dairy heifers.

This study aimed to determine effects of exposure of recipient dairy heifers to heat stress (THI ≥ 73) during the oestrous cycle coinciding with embryo transfer (ET) on the risk of pregnancy establishment after transfer of in vivo produced embryos. Recipients exposed to THI values ≥73 during Days zero (recipient estrus), 7 (day of ET), 14 (seven days after ET), 15 and 16 (maternal recognition of pregnancy) of the ET cycle were considered as heat-stressed heifers (n = 254), while heifers in the control group (n = 470) were not exposed to THI ≥ 73 at any of the previous days. Results revealed no significant effects of any of the investigated factors on the risk of pregnancy following ET. However, the mean THI above 77 was associated with a drastic numerical decrease in PR/ET (36.63%), when compared to a mean THI 72 (78.78%). In addition, PR/ET after transfer of second- and third-grade embryos were numerically lower in heat-stressed recipients, compared with first-grade embryos (41.17% vs. 56.36%, respectively). Our findings confirmed that transfer of blastocysts was associated with numerically higher PR/ET in heat-stressed and control recipients, as compared to morula stage. Interestingly, PR/ET tended to be higher when sexed embryos were transferred to the control recipients compared with heat-stressed ones. In conclusion, PR/ET in dairy heifers was not significantly affected by heat stress during critical windows of their oestrous cycle coinciding with ET, whereas transfer of sexed embryos gives lower results under conditions of heat stress.

Detection of Giardia intestinalis assemblages A and B among children from three villages in the West Delta region, Egypt using assemblage specific primers.

Giardia intestinalis is a common diarrheagenic parasite infecting children globally. It has been classified into eight morphologically identical but genetically distinct genotypes. Human infection is mainly associated with A and B assemblages with variable geographical distribution. The present work aimed to study the epidemiology of assemblages A and B in children inhabiting different areas in Lower Egypt. Stool samples were collected from 315 children and examined microscopically for parasitic infections. Giardia positive samples were genotyped using tpi assemblage specific primers. The prevalence of Giardia was 18.1% among the examined children. Mixed assemblages A and B was more common (47.4%) than single assemblage B (36.8%) or A (15.8%). The distribution of different genotypes was significantly associated with the residence area, animal contact, and handwashing habits. A non-significant association was observed between Giardia assemblages and the clinical manifestations. Assemblage B is the predominant genotype among Egyptian children. The distribution of different Giardia assemblages is strongly associated with the studied area and the habits of its people.

Chlorella vulgaris ameliorates sodium nitrite-induced hepatotoxicity in rats.

The current was conducted to evaluate the ameliorating effect of Chlorella vulgaris (CV) extract against sodium nitrite-induced hepatotoxicity in rats. Forty-five rats were allocated randomly into 5 groups (n = 9). Group I (GI), control group: orally gavaged with normal saline daily. Group II (GII): orally gavaged with CV extract (70 mg/kg BW) for 3 months. Group III (GIII): orally gavaged with sodium nitrite (80 mg/kg BW) for 3 months. Group IV (GIV): received sodium nitrite as GIII and CV extract as GII simultaneously for 3 months. Group V (GV): received CV extract as GII and then, sodium nitrite as in GIII from the end of first month until the end of the experiment. Sodium nitrite significantly increased the activities of serum alanine aminotransferase, aspartate aminotransferase, and serum concentrations of tumor interleukin 1-ß and necrosis factor α. In addition, it increased concentrations of malondialdehyde and nitric oxide and expression level of caspase-3 in the hepatic tissue. However, it decreased activities of hepatic glutathione peroxidase, catalase, and superoxide dismutase and induced degenerative and necrotic changes in hepatic tissues. In contrast, CV extract administration modulated sodium nitrite-induced inflammation, oxidative stress, and alteration in hepatic tissue function and architecture. This study indicated that CV extract modulated sodium nitrite-induced hepatic toxicity through decreasing oxidative stress and inflammation and enhancing antioxidant enzyme activities in hepatic tissue of rats.

Methanolic extract of Chlorella vulgaris protects against sodium nitrite-induced reproductive toxicity in male rats.

The current study aimed to investigate the protective potential of Chlorella Vulgaris (CV) extract against the reproductive dysfunction induced by sodium nitrite toxicity. Forty-five male Wistar albino rats were assigned into five groups (n = 9). Control group received normal saline orally for 3 months, CV-treated: administered CV extract (70 mg/kg.BW) orally for 3 months, sodium nitrite-treated: received sodium nitrite (80 mg/kg.BW) orally for 3 months, co-treated: simultaneously received CV along with sodium nitrite treatment, orally, daily for 3 months, and CV-pre-treated: pre-treated with CV extract for 4 weeks followed by simultaneous treatment with sodium nitrite and CV extract for additional 8 weeks. Treatment with sodium nitrite significantly decreased serum testosterone and follicle-stimulating hormone concentrations, sperm count, motility, and viability. Besides, it decreased testicular superoxide dismutase and glutathione peroxidase activities while increased malondialdehyde concentration. This effect of sodium nitrite was associated with degenerative, necrotic, vascular, and inflammatory changes in testicular tissues. Treatment of sodium nitrite-intoxicated rats with CV in co-treated and pre-treated groups significantly prevented sodium nitrite-induced alterations of sperm parameters, hormonal concentrations, testicular oxidative-antioxidant status, and histological architecture. This study indicates that CV extract ameliorates the reproductive dysfunction induced by sodium nitrite toxicity via improving reproductive hormonal levels and testicular antioxidant activities.

Validation of electrical velocimetry in resuscitation of patients undergoing liver transplantation. Observational study.

Major hemodynamic changes are frequently noted during liver transplantation (LT). We evaluated the performance of electrical velocimetry (EV) as compared to that of TEE in SV optimization during liver transplantation. This was an observational study in 32 patients undergoing LT. We compared SV values measured simultaneously by EV (SVEV) and TEE (SVTEE) at baseline 30 min after induction, at the end of dissection phase, 30 min after anhepatic phase, 30 min after reperfusion. We also evaluated the reliability of EV to track changes In SV before and after 49 fluid challenges. Finally, the SV variation (SVV) and pulse pressure variation (PPV) were tested as predictors for volume responsiveness, defined as an increase in SV ≥ 10% after 250 ml of colloid. For 112 paired SV data, the overall correlation was 0.76 and bias (limits of agreement) 0.3 (- 29 to 29) ml percentage error 62%. The EV was able to track changes in SV with a concordance rate of 97%, and a sensitivity and specificity of 93% to detect a positive fluid challenge. The AUC values (with 95% confidence intervals) for SVV and PPV were 0.68 (0.52-0.83) and 0.72 (0.57-0.86), respectively, indicating low predictive capacity in these setting. The absolute values of SV derived from EV did not agree with SV derived from TEE. However, EV was able to track the direction of changes in SV during hemodynamic management of patients undergoing liver transplantation.Clinical trial registration: Clinicaltrials.gov Identifier: NCT03228329 prospectively Registered on 13-July-2017.

Development and optimization of a novel drug free nanolipid vesicular system for treatment of osteoarthritis.

OBJECTIVE: The goal of this study is to improve the transdermal delivery of phosphatidylcholine (PC) via constructing a novel nanolipid vesicular system (NLVS) with high level of permeability through the stratum corneum (SC). SIGNIFICANCE: In our study, a novel drug free NLVS was developed. The system depends on PC boundary cartilage lubrication to relieve osteoarthritic pain without developing gastrointestinal problems associated with anti-inflammatory drug. MATERIALS AND METHODS: A full two-level (23) factorial design is applied to optimize the quality of the prepared NLVS. The selected independent variables are the concentration of PC, the concentration of edge activator (EA), and EA type. The developed NLVS was evaluated for in-vitro, ex-vivo as well as in-vivo efficacy in rat animal model. RESULTS: Based on the factorial design, the selected formulation variables significantly affect the tested responses. The prepared NLV formulations have a particle size (PS)in the range of 10.34 to 496.3 nm, polydispersity index (PdI) values less than one, and negative zeta potential (ZP) range of -1.42 to -32.01 mV. In-vitro and ex-vivo study results reveal that the designed NLVS is effective in sustaining PC release and enhancing its transdermal permeation over 24 h. The optimal permeation flux through ex-vivo study is 0.415 mg/cm2/h following zero-order kinetics. Moreover, in-vivo study of the optimized formulations demonstrated remarkable reduction in inflammatory mediators associated with osteoarthritis (OA). CONCLUSION: The results indicate that the optimized drug free NLVS significantly augment transdermal delivery of PC and have a potential role in treatment of OA without the risk of systemic side effects.

Ameliorative effect of pumpkin seed oil against emamectin induced toxicity in mice.

The current study was conducted to evaluate the toxic effects of emamectin insecticide in mice and the possible protective effect of pumpkin seed oil. Treated mice received emamectin benzoate in the diet at 75-ppm for 8 weeks, while another group of animals received emamectin in addition to pumpkin seed oil at a dose of 4 ml/kg. Biochemical analysis of MDA, DNA fragmentation, GSH, CAT and SOD was performed in liver, kidney and brain as oxidant/antioxidant biomarkers. In addition, gene expression of CYP2E1 and Mgst1 and histopathological alterations in these organs were evaluated. Emamectin administration induced oxidative stress in liver and kidney evidenced by elevated levels of MDA and percentage of DNA fragmentation with suppression of GSH level and CAT and SOD activities. Brain showed increase of MDA level with inhibition of SOD activity. Relative expressions of CYP2E1 and Mgst1 genes were significantly elevated in both liver and kidney. Emamectin produced several histopathological changes in liver, kidney and brain. Co-administration of pumpkin seed oil produced considerable protection of liver and kidney and complete protection of brain. In conclusion, pumpkin seed oil has valuable value in ameliorating the toxic insult produced by emamectin in mice.

Clinical, pathological, and molecular data concerning Coenurus cerebralis in Sheep in Egypt.

This article contains information related to a recent study "Prevalence and Identity of Taenia multiceps cysts "Coenurus cerebralis" in Sheep in Egypt" (Amer et al., 2017) [1]. Specifically, affected sheep showed neurological disorders manifested as depression, head shaking and circling, altered head position, incoordination and paralysis in some cases. Brain-derived cysts were molecularly identified by PCR-sequence analysis at mitochondrial 12S rRNA gene marker. Cyst-induced pathological changes included degenerative changes and demyelination in brain tissue, infiltration of lymphocytes and histiocytes. Cystic fluids were biochemically analyzed for protein, lipids and electrolytes. The data of this study provides more understanding on phylogeny, epidemiology and pathology of coenurosis in sheep.

Prevalence and Identity of Taenia multiceps cysts "Coenurus cerebralis" in Sheep in Egypt.

Coenurosis is a parasitic disease caused by the larval stage (Coenurus cerebralis) of the canids cestode Taenia multiceps. C. cerebralis particularly infects sheep and goats, and pose a public health concerns. The present study aimed to determine the occurrence and molecular identity of C. cerebralis infecting sheep in Egypt. Infection rate was determined by postmortem inspection of heads of the cases that showed neurological manifestations. Species identification and genetic diversity were analyzed based on PCR-sequence analysis of nuclear ITS1 and mitochondrial cytochrome oxidase (COI) and nicotinamide adenine dinucleotide dehydrogenase (ND1) gene markers. Out of 3668 animals distributed in 50 herds at localities of Ashmoun and El Sadat cities, El Menoufia Province, Egypt, 420 (11.45%) sheep showed neurological disorders. Postmortem examination of these animals after slaughter at local abattoirs indicated to occurrence of C. cerebralis cysts in the brain of 111 out of 420 (26.4%), with overall infection rate 3.03% of the involved sheep population. Molecular analysis of representative samples of coenuri at ITS1 gene marker showed extensive intra- and inter-sequence diversity due to deletions/insertions in the microsatellite regions. On contrast to the nuclear gene marker, considerably low genetic diversity was seen in the analyzed mitochondrial gene markers. Phylogenetic analysis based on COI and ND1 gene sequences indicated that the generated sequences in the present study and the reference sequences in the database clustered in 4 haplogroups, with more or less similar topologies. Clustering pattern of the phylogenetic tree showed no effect for the geographic location or the host species.

Sequential protocol biopsies from renal transplant recipients show an increasing expression of active TGF beta.

Chronic allograft nephropathy (CAN) is a major cause of graft loss after renal transplantation. Implicated in the pathogenesis of this complication is overproduction of the cytokine transforming growth factor beta (TGF beta). In this study we measured changes in CAN's expression in stable patients early after transplantation, and studied links with established risk factors for CAN, such as delayed graft function, acute rejection, and cyclosporine exposure. We took biopsies from 40 renal allografts at time of transplantation (pre-perfusion), and then, using ultrasound guidance, at 1 week and 6 months after transplantation. An immunofluorescence technique was used to stain sections for active TGF beta. These were then assessed by semi-quantitative scanning laser confocal microscopy. There was very little variation in active TGF-beta expression among patients in their pre-perfusion biopsies. Expression had increased by 1 week and then very significantly by 6 months ( P<0.0001). Patients who suffered delayed graft function had increased TGF-beta expression at both time points. There was no difference regarding donor type, acute rejection, and immunosuppressive drug (cyclosporine or tacrolimus). There was no correlation between the amount of TGF-beta expression at any time-point and isotope glomerular filtration rate (GFR) at 12 months. This study demonstrated that in a group of stable renal allograft recipients, TGF-beta expression in the kidney increased after transplantation. As the study used protocol biopsies, this increase is unlikely to be due to acute events, and probably represents a genuine increase.