Activities of pumpkin seed oil against Biomphalaria alexandrina snails and the infective stages of Schistosoma mansoni with special emphasis on genotoxic and histopathological alterations.

Schistosomiasis is a serious health issue in tropical regions, and natural compounds have gained popularity in medical science. This study investigated the potential effects of pumpkin seed oil (PSO) on Biomphalaria [B.] alexandrina snails (Ehrenberg, 1831), Schistosoma [S.] mansoni (Sambon, 1907) miracidium, and cercariae. The chemical composition of PSO was determined using gas chromatography/mass spectrometry. A bioassay was performed to evaluate the effects of PSO on snails, miracidia, and cercariae. The results showed no significant mortality of B. alexandrina snails after exposure to PSO, but it caused morphological changes in their hemocytes at 1.0 mg/ml for 24 hours. PSO exhibited larvicidal activity against miracidia after 2 hours of exposure at a LC50 of 618.4 ppm. A significant increase in the mortality rate of miracidia was observed in a dose- and time-dependent manner, reaching a 100% death rate after 10 minutes at LC90 and 15 minutes at LC50 concentration. PSO also showed effective cercaricidal activity after 2 hours of exposure at a LC50 of 290.5 ppm. Histological examination revealed multiple pathological changes in the digestive and hermaphrodite glands. The PSO had genotoxic effects on snails, which exhibited a significant increase [p≤0.05] in comet parameters compared to the control. The findings suggest that PSO has potential as a molluscicide, miracidicide, and cercaricide, making it a possible alternative to traditional molluscicides in controlling schistosomiasis.

Binary and ternary Cu(II) complexes of pregabalin with excitatory and inhibitory neurotransmitters and their antiepileptic effect.

Antiepileptic drug (Preg) and neurotransmitters (Gpn, Gly, Asp, Glu, Ser, GABA and Ade) were used to synthesis a series of ternary Cu2+ complexes. Surface morphology and chemical composition of the complexes were using studied SEM and EDX spectra. Purity, molecular weight and formulae of the complexes were determined from GC-MS spectra and elemental analysis. XRD data and N-Treor implemented in Expo2014 computer program reveal monoclinic crystal system with space group P1 2/c 1 and P 1 21 1 of the complexes. IR spectra exhibited that Preg, Gpn and GABA coordinated to the Cu2+ as monodentate ligand through COOH whereas the amino acids bonded through the -NH2, COO- groups. UV-Vis spectra and magnetic moment values indicated pseudo tetrahedral stereochemistry. ESR spectra showed that the complexes have isotropic and axial structures with dx2-y2 ground state. TGA, DTG and DTA confirm the suggested structure and mechanism for thermal decomposition was suggested. Kinetic and thermodynamic parameters were calculated using Coats-Redfern equation. The complexes [Cu(Preg)(Ser)Cl] and [Cu(Preg)(Ade)Cl2] showed greater anticonvulsant activity compared against PTZ-induced seizures in male Albino mice. Recorded latency time for the complexes [Cu(Preg)(Ser)Cl] and [Cu(Preg)(GABA)(OH)Cl].2H2O was longer than that recorded with Preg indicating higher anticonvulsant effect.

Staphylococcus pseudintermedius Sbi paralogs inhibit complement and bind IgM, IgG Fc and Fab.

The success of staphylococci as pathogens has been attributed, in part, to their ability to evade their hosts' immune systems. Although the proteins involved in evasion have been extensively studied in staphylococci affecting humans little characterization has been done with Staphylococcus pseudintermedius, an important cause of pyoderma in dogs. Staphylococcus aureus binder of immunoglobulin (Sbi) interferes with innate immune recognition by interacting with multiple host proteins. In this study, a S. pseudintermedius gene that shares 38% similarity to S. aureus Sbi was cloned from S. pseudintermedius strains representative of major clonal lineages bearing two paralogs of the protein. Binding of immunoglobulins and Fab and Fc fragments as well as interaction with complement was measured. S. pseudintermedius Sbi protein bound IgG from multiple species and canine complement C3, neutralized complement activity and bound to canine IgM and B cells. Evidence from this work suggests Sbi may play an important role in S. pseudintermedius immune evasion.

Molecular characterization of virulence and antibiotic resistance genes among Salmonella serovars isolated from broilers in Egypt.

This study was carried out to investigate the resistance to some antimicrobial groups among Salmonellae isolated from broilers in Egypt. The prevalence of some virulence and resistance genes among the recovered multidrug resistant (MDR) isolates was also scrutinized. A total of 55 (15·6%) Salmonella isolates were recovered from 353 different samples (liver, yolk sac, gall bladder and caecum), gathered from apparently healthy and diseased broilers suffered from diarrhoea, dehydration and respiratory distress. Thirty Salmonellae strains were serotyped into Salmonella Enteritidis, Salmonella Infantis, Salmonella Kentucky, Salmonella Maloma, Salmonella Bardo, Salmonella Gdansk, Salmonella Typhimurium and Salmonella Blegdame. The resistance pattern of all Salmonella isolates was constructed and 15 MDR Salmonella isolates were subsequently examined for the presence of virulence (invA, ompA and stn) and resistance (qnrS, qnrA, blaTEM and blaCTX ) genes. Of note, invA, ompA and stn virulence genes and blaTEM and qnrS resistance genes were found in all examined isolates. On the other hand, the qnrA gene detection frequency was 20%, whereas blaCTX was not detected at all. Our findings emphasize the wide spread of antimicrobial resistance genes in Salmonella isolates and the importance of effective control measures for the disease. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights the possible emergence of widespread resistance to some antimicrobials among Salmonellae isolated from broilers in Egypt. The results also reveal the prevalence of some virulence (invA, ompA and stn) and resistance (qnrS, qnrA, blaCTX and blaTEM ) genes among the recovered multidrug resistant isolates. Clearly, our data emphasize that antimicrobial resistance genes are widely spread in Salmonella isolates which is crucial for developing novel methods for controlling such disease problem of zoonotic concern.

Identification, Cloning, and Characterization of Staphylococcus pseudintermedius Coagulase.

Coagulase activation of prothrombin by staphylococcus induces the formation of fibrin deposition that facilitates the establishment of infection by Staphylococcus species. Coagulase activity is a key characteristic of Staphylococcus pseudintermedius; however, no coagulase gene or associated protein has been studied to characterize this activity. We report a recombinant protein sharing 40% similarity to Staphylococcus aureus coagulase produced from a putative S. pseudintermedius coagulase gene. Prothrombin activation by the protein was measured with a chromogenic assay using thrombin tripeptide substrate. Stronger interaction with bovine prothrombin than with human prothrombin was observed. The S. pseudintermedius coagulase protein also bound complement C3 and immunoglobulin. Recombinant coagulase facilitated the escape of S. pseudintermedius from phagocytosis, presumably by forming a bridge between opsonizing antibody, complement, and fibrinogen. Evidence from this work suggests that S. pseudintermedius coagulase has multifunctional properties that contribute to immune evasion that likely plays an important role in virulence.

Genetic basis of resistance waves among methicillin resistant Staphylococcus aureus isolates recovered from milk and meat products in Egypt.

Antimicrobial resistance of methicillin-resistant Staphylococcus aureus (MRSA) poses a serious problem for clinicians worldwide. The present study attempted to evaluate the susceptibility patterns of MRSA to various antimicrobials and the prevalence of inducible clindamycin resistance as well as the relevant antibiotic and antiseptic resistance genes among these isolates. Totally, 40 MRSA isolates were recovered from examined milk and meat product samples (18.60%). Multi-drug resistance (MDR) was remarkably observed among 85% of these isolates. There was a good correlation between phenotypic determination of methicillin, amoxicillin/clavulinic acid and tetracycline resistances and PCR detections of mecA, blaZ and tet(K) genes, respectively, but norA gene was not detected in the four ciprofloxacin resistant isolates. Although, 55% of MRSA expressed resistance to benzalkonium chloride (BC), neither qacA/B nor smr gene was detected. Of 20 isolates exhibiting erythromycin- clindamycin discordant resistance pattern, 8 displayed positive double disk diffusion (D-zone) test denoting inducible macrolide-lincosamide-streptogramin B (MLSB) resistance phenotype with the inducibly expressed erm(A) and erm(C) genes in 87.5% of these isolates. Besides, the remaining 12 isolates showed MS phenotype (resistant to macrolides and type B streptogramins only) with a variety of erm(A), mph(C), msr(A) or a combination of these genes including erm(C). Finally, the constitutive MLSB phenotype with the constitutive expression of erm(A), erm(B) and erm(C) genes was comprised in 2 isolates with higher minimum inhibitory concentration (MIC) values for erythromycin (512 and 1024 µg/ml) and clindamycin (16 and 32 µg/ml). These findings suggested the importance of monitoring the evolution of MRSA resistance.

Molecular versus conventional culture for detection of respiratory bacterial pathogens in poultry.

Acute respiratory tract infections are leading causes of morbidity in poultry farms allover the world. Six pathogens; Escherichia coli, Mycoplasma gallisepticum, Staphylococcus aureus, Pasteurella multocida, Mannheimia haemolytica and Pseudomonas aeruginosa were involved in respiratory infections in poultry. Herein, conventional identification procedures and polymerase chain reaction (PCR) were applied for detection of the most common respiratory bacterial pathogens in clinical specimens of poultry obtained from 53 Egyptian farms with various respiratory problems and the results were compared statistically. The analyzed data demonstrated a significantly higher rate of detection of the most recovered microorganisms (P<0.05) by PCR comparing to classical culture procedures. Further, multiplex PCR could detect E. coli, M. gallisepticum, S. aureus and Ps. aeruginosa in a single reaction, however, M. haemolytica was reported in a uinplex system. According to PCR results, the most commonly recorded bacterial pathogens in examined poultry farms were E. coli and Ps. aeruginosa (54.71% each), followed by M. haemolylica (35.85%) and M. gallisepticum (20.75%). In conclusion, PCR assay offered an effective alternative to traditional typing methods for the identification and simultaneous detection of the most clinically relevant respiratory pathogens in poultry.

Extended-Spectrum ß-Lactamases and/or Carbapenemases-Producing Enterobacteriaceae Isolated from Retail Chicken Meat in Zagazig, Egypt.

OBJECTIVES: The aim of the present study was to determine the prevalence and to characterize extended-spectrum ß-lactamases- and/or carbapenemases-producing Enterobacteriaceae among Enterobacteriaceae isolated from retail chicken meat in Zagazig, Egypt. METHODS: One hundred and six Enterobacteriaceae isolates were collected from retail chicken meat samples purchased in Zagazig, Egypt in 2013. Species identification was done by MALDI-TOF MS. Screening for ESBL-E was performed by inoculation of isolates recovered from meat samples onto the EbSA (Cepheid Benelux, Apeldoorn, the Netherlands) selective screening agar. ESBL production was confirmed by combination disc diffusion test with clavulanic acid (Rosco, Taastrup, Denmark). Carbapenemases production was confirmed with double disk synergy tests. Resistance genes were characterized by PCR with specific primers for TEM, SHV, and CTX-M and carbapenemases (KPC, NDM, OXA-48, IMP and VIM). PCR products of CTX-M genes were purified and sequenced. Phylogenetic grouping of E. coli was performed by a PCR-based method. RESULTS: Of these 106 isolates 69 (65.09%) were ESBL producers. Twelve (11.32%) of these isolates were also phenotypically class B carbapenemases producer. TEM genes were detected in 61 (57.55%) isolates. 49 (46.23%) isolates harbored CTX-M genes, and 25 (23.58%) carried genes of the SHV family. All CPE belonged to the NDM group. The predominant CTX-M sequence type was CTX-M-15 (89.80%). The majority (80%) of the ESBL-EC belonged to low virulence phylogroups A and B1. CONCLUSIONS: This is the first study from Egypt reporting high rates of ESBLs and carbapenemases (65.09% and 11.32%, respectively) in Enterobacteriaceae isolated from retail chicken meat. These results raise serious concerns about public health and food safety as retail meat could serve as a reservoir for these resistant bacteria which could be transferred to humans through the food chain.

Extended-Spectrum ß-Lactamase- and Carbapenemase-Producing Enterobacteriaceae Isolated from Egyptian Patients with Suspected Blood Stream Infection.

OBJECTIVES: The aim of the study was to investigate the prevalence of extended-spectrum ß-lactamase and carbapenemase production among Enterobacteriaceae isolated from Egyptian patients with suspected blood stream infection. METHODS: Ninety-four Enterobacteriaceae blood culture isolates from Egyptian patients with suspected blood stream infection were collected, one isolate per patient. Identification of bacterial isolates was performed with MALDI-TOF (MS-based Vitek MS system, bioMerieux). Screening for ESBLs and carbapenemases production was done with the Vitek 2 system (bioMérieux). ESBL production was confirmed using the combined disk diffusion method for cefotaxime, ceftazidime, and cefepime, all with and without clavulanic acid (Rosco). Real-time PCR and sequencing were used to characterize the resistance genes. The phylogenetic groups of E. coli were identified by a PCR-based method. RESULTS: Of the 94 Enterobacteriaceae isolates 46 (48.93%) showed an ESBL phenotype. One Enterobacter spp isolate was ESBL-producer and meropenem-resistant. The genetic analysis showed that CTX-M was present in 89.13% (41/46) of the ESBL-producing Enterobacteriaceae, whereas TEM and SHV were detected in 56.52% (26/46) and 21.74% (10/46) respectively (47.83%) of the ESBL-producing isolates were multidrug resistant (MDR). Eleven out of 30 ESBL-producing E-coli isolates were assigned to phylogroup B2, followed by groups B1 (8 isolates), A (6 isolates) and D (5 isolates). CONCLUSIONS: The high ESBL-E rates (48.93%) found in this study together with the identification of one carbapenem-resistant Enterobacter spp isolate is worrisome. Our results indicate that systems for monitoring and detection of ESBL-producing bacteria in Egyptian hospitals have to be established. Also strict hospital infection control policies with the restriction of the consumption of extended-spectrum cephalosporins are necessary.

Serological investigation of horse sera for antibodies against mycoplasmas and acholeplasmas.

Sera from horses with respiratory disease (RD) have been investigated using the complement fixation test, indirect hemagglutination test, enzyme immune assay, and the metabolic inhibition test, and sera from mares after abortion, using the complement fixation test, indirect hemagglutination test and enzyme immune assay, for antibodies against Mycoplasma equirhinis, M subdolum, M. equigenitalium, M. pulmonis, M. felis, Acholeplasma laidlawii, A. hippikon and A. equifetale. Antibodies were found against all mycoplasma and acholeplasma species tested, more often against acholeplasmas. The antibody pattern was quite similar for horses with RD and for mares after abortion. The results of the four serological tests performed showed only a limited correlation and the percentage of sera with antibodies detected by the four tests used differed widely.

[Investigation of mare sera for antibodies against acholeplasmas and mycoplasmas with ther enzyme linked immunosorbent assay (ELISA) (author's transl)]. / Untersuchung von Stutenseren auf Antikörper gegen Acholeplasmen und Mykoplasmen mit dem Enzyme Linked Immunosorbent Assay (ELISA).

After abortion sera were taken from 58 thoroughbred and other mares of the northwestern part of Germany and investigated by ELISA (enzyme linked immuno-sorbent assay) for antibodies against Mycoplasma equirhinis, M. subdolum, M. equigenitalium, M. pulmonis, M. felis, Acholeplasma laidlawii, A. hippikon, and A. equifetale. Reactions at serum dilutions of 1:32 and higher were considered as positive. At serum dilution 1:32 no antibodies were found in 11 sera. The remaining sera showed antibodies against one or more of the mycoplasma antigens investigated. The number of multiple reactions decreased with an increasing dilution of the sera. Titers were found between 1:32 and 1:256. In one case a titer of 1:2048 against M. equigenitalium antigen was found. Most often antibodies against A. laidlawii were observed i.e. in 37 sera. These antibodies also showed the highest titers. Only 3 sera contained antibodies against A. hippikon. Antibodies against M. felis and A. equifetale were found in 26 sera. Between 10 and 15 sera showed antibodies against the remaining mycoplasma species.