Molecular characterization of haemagglutinin-neuraminidase gene among virulent Newcastle disease viruses isolated in Iran.

BACKGROUND: Virulent Newcastle disease virus (vNDV) causes great economic losses to the poultry industry throughout the world. Despite the endemicity of Newcastle disease (ND) and occurrence of recurrent outbreaks, the nature and genetic features of circulating NDV strains in Iran are largely unknown. Aims: This study was conducted to characterize 13 NDV isolates obtained from different outbreaks in various regions of Iran during 1999-2000 by sequencing and phylogenetic analysis of complete coding sequences of haemagglutinin-neuraminidase (HN) gene. METHODS: All isolates were analyzed based on the previously determined in vivo pathogenicity indices and amino acid (aa) sequences of fusion (F) protein cleavage site (FPCS). RESULTS: Phylogenetic analysis based on the HN gene coding region revealed a very close relationship of these viruses with the recently defined genotype XIII, and more specifically, subgenotype XIIIa viruses. Analysis of HN gene nucleotide (nt) sequences revealed that all studied isolates encode for a protein length of 571 aa and there is no C-terminal extension on HN aa sequences. Sequence analysis revealed multiple aa residue substitutions at antigenic sites or neutralizing epitopes on the HN glycoprotein of studied viruses compared with commonly used vaccinal strains. CONCLUSION: In this study, molecular characterization of vNDV isolates, obtained from commercial poultry farms in Iran, were conducted through complete sequencing and analysis of HN gene. Isolation and molecular characterization of further NDV isolates from other parts of Iran and from neighboring countries in the region will be helpful to identify the nature and origin of indigenous viruses.

Bleaching effects of massecuite on some properties of crystallized sugar beet and milk chocolate bars produced.

Four kinds of bleached, unbleached, second and third crystal sugars (BCS, UCS, SCS and TCS) were made from different massecuites in a sugar-beet factory, and their physiochemical (polarization, invert sugar, colorants, pH, ash and SO2), microbiological and functional properties were measured. While the polarization of UCS, SCS and TCS were lower than BCS; their invert sucrose, colorants, pH and ash contents were significantly higher than BCS. The phenols and betaine of BCS, UCS, SCS, and TCS were 144, 401, 384 and 673 (mg/100 g); and 244, 791, 4662, and 6589 (mg/100 g); respectively. Whereas the phenol of milk chocolate bars (MCB) made with UCS, SCS, and TCS were only 10% higher than MCB completed with BCS; their betaine contents were substantially (up to 16 times) higher than the ones finished with BCS. Sensory evaluation showed that the MCB prepared with three sugars including UCS, SCS and TCS had significantly higher glossiness, brittleness, flavor and mouth feel than those made with BCS. The greater colorants, ash content and inverted sugars of UCS, SCS and TCS (in comparison with BCS) made considerable improvements in the glossiness, flavor and brittleness of MCB, respectively. BCS had 8 ppm of toxic sulfur; whereas, UCS, SCS and TCS had no detectable sulfur and significantly higher beneficial copper content than BCS. No pathogenic microorganism were detected in UCS, SCS, TCS or their subsequent MCB. Our results highly recommend using UCS, SCS and TCS instead of BCS in food products (such as MCB) due to their higher health benefits.

Protective effect of pravastatin on doxorubicin-induced hepatotoxicity.

OBJECTIVE: The present study was designed to investigate the possible protection of pravastatin against hepatic oxidative stress and dysfunctions induced by doxorubicin in rats. BACKGROUND: Statins have beneficial effects on oxidative stress and inflammation. METHODS: Male Sprague-Dawley rats were divided into four groups. Control group (received saline orally), Group 2 received pravastatin (20 mg/kg, i.p. for 15 days), Group 3 received single dose doxorubicin (15 mg/kg, i.p.), Group 4 was treated with pravastatin (20 mg/kg, i.p.) daily from 5 days before to 10 days after injection of doxorubicin (15 mg/kg, i.p.). Hepatic toxicity was estimated by biochemical parameters and oxidative stress and histopathological studies. RESULTS: Administration of doxorubicin indicated an increase in ALT, AST, ALP, TG, cholesterol, LDL and total bilirubin levels (p < 0.01). Doxorubicin caused a reduction in HDL and albumin levels (p < 0.01) as well as superoxide dismutase, glutathione peroxidase and catalase activities (p < 0.05) with a concomitant increase in liver malondialdehyde (p < 0.05) and liver damage (p < 0.001). Pravastatin reduced the scale liver injury (p < 0.001) and protected liver functions and other biochemical parameters (p < 0.01). Increase in malondialdehyde level associated with a reduction in antioxidant activities in the doxorubicin group was attenuated by pravastatin treatment (p < 0.05). CONCLUSION: Results indicated that pravastatin has a protective effect on the liver against doxorubicin-induced hepatotoxicity in rats (Tab. 3, Fig. 2, Ref. 34).

Detection of Mycoplasma agalactiae in Small Ruminants of Southeast Iran.

Agalactia is an infectious and contagious disease of small ruminants caused by Mycoplasma agalactiae (M. agalactiae). Although different microorganism strains contribute to this disease, M. agalactiae is known as the most prominent causative agent. Therefore, this study aimed to investigate the rate of M. agalactiae involvement in contagious agalactia in the southeast region of Iran. Sampling was performed from milk, conjunctiva, ear lesions, and joints exudate of suspicious sheep and goat flocks according to the reports of Iran Veterinary Organization. The presence of Mycoplasma and its species, namely M. agalactiae, was evaluated through microbial culture and polymerase chain reaction (PCR) techniques. The detected microorganisms were confirmed to be Mycoplasma and M. agalactiae by the PCR amplification of 16S rRNA and lipoprotein target genes. According to the findings of present study, 14.8% and 36.0% of the samples were diagnosed as positive for Mycoplasma by culture and PCR, respectively. Moreover, the incidence of M. agalactiae was determined as 6.1% using the specific PCR method. Therefore, it is recommended to identify the other species of Mycoplasma in small ruminant samples involved with contagious agalactiae disease.

Isolation and Detection of Mycoplasma agalactiae from Semen Samples of Goats.

Contagious agalactia (CA) is a highly infectious disease of goats and sheep, and is a form of Mycoplasmosis, which is usually enzootic. Since Mycoplasma agalactiae (M. agalactiae) is the main cause of this disease in goats, the aim of this study was to isolate and detect M. agalactiae from semen of goat bucks. Thirty-nine semen samples were collected from goat bulks, and all samples were cultured in PPLO broth medium supplemented for M. agalaciae isolation. The bacteria DNAs were extracted from clinical samples and the PCR assay was applied to detect Mycoplasma genus and M. agalactiae species using specific primers, which amplified a 163bp fragment in 16SrRNA gene and a 375bp fragment in lipoprotein gene. The PCR evaluations were performed for both the clinical samples and the cultures. Out of the 39 samples, 29 (74.3%) of the cultures were shown positive and typical Mycoplasma colonies grew on PPLO agar, which could be considered as the diagnostic method. In addition, 38 (97.4%) samples had positive PCR results for Mycoplasma genus and six (15.3%) of the samples were shown to be positive using PCR for M. agalactiae as the diagnostic method. In the present study, M. agalactiae was detected in semen of goat bulks for the first time in Iran. Therefore, it is recommended to concern semen as one of the significant sources for this pathogen and the possibility for transmission to the female goats through semen is highlighted. Moreover, presence of this microorganism in semen could be involved in infertility of goat population.

Co-circulation of genetically distinct groups of avian paramyxovirus type 1 in pigeon Newcastle disease in Iran.

Pigeons are considered as one of the major natural reservoirs in the epidemiology of Newcastle disease (ND). In this study, the partial sequence of fusion protein gene of 17 pigeon-origin ND viruses (NDVs) isolated during 2012-2013 in Iran was analysed. Since the studied isolates showed F0 protein cleavage sites compatible with velogenic NDVs, all were considered as virulent NDVs. Two isolates carried 112RRQKRF117 as the cleavage site motif, whereas the rest demonstrated 112KRQKRF117 motif which just recently has been reported among Iranian virulent NDVs. Phylogenetic analysis divided all these diverse isolates in two distinct clusters within class II genotype VI. Based on the partial fusion protein gene sequence, 15 out of 17 isolates showed the highest genetic identity to subgenotype VIb/2 and the other two isolates were placed in a distinct genetic group of genotype VI. Based on recent findings, at least two different sublineages of genotype VI are causing the ND outbreaks in the pigeon population and are circulating simultaneously along with virulent NDVs of genotype VII in various species in Iran. The continuing circulation of a diverse group of virulent NDVs as an enzootic in widespread species such as pigeon can cause outbreaks in commercial poultry flocks and also failure in controlling programmes. Therefore, the constant monitoring and awareness of the virus characteristics should be considered in controlling programmes against ND in Iran.

Effects of sulfur water extraction on anthocyanins properties of tepals in flower of saffron (Crocus sativus L).

A sulfur solution with different metabisulfite concentrations (100, 400, 700, 1,000 and 2,000 ppm) was used to extract anthocyanins from saffron tepals. The extraction process was compared with acidified ethanol solution at similar extraction times of 20, 40, 60, 120, and 180 min at 40 °C. The recovery of anthocyanins with sulfur solution was higher than ethanol extraction and reached to 700 mg anthocyanins/100 g, when the sulfur concentration and extraction time were 700 ppm and 60 min, respectively. HPLC analysis showed that anthocyanins extracted with sulfur solution followed by partial desulfurization and reducing sulfur content (to less than 250 ppm) had around 100 % more cyanidin 3 glucosides and 100 % less pelargonidin 3,5 glucosides in comparison with ethanol extraction. Additionally, the color of low-sulfured anthocyanins had more saturation (chroma), less lightness, and more stability than the one extracted with ethanol solution. While monomeric and polymeric anthocyanins extracted with sulfur solution had less than 1 % changes after 3 h extraction time, they had more than 12 % changes when they extracted with alcoholic solution at similar conditions. Overall, the sulfur method had a potential to extract stable anthocyanins from waste and discarded saffron tepals in aqueous solvent, and with higher quantity and quality (more attractive color) than conventional ethanol extraction method.

Multidetector computed tomography (MDCT) in the diagnosis of pulmonary embolism: interobserver agreement among radiologists with varied levels of experience.

PURPOSE: To assess the interobserver variability of radiologists with varied levels of experience in the interpretation of multidetector computed tomography (MDCT) pulmonary angiographies. MATERIAL AND METHODS: Review of CT pulmonary angiographies performed on patients included in a diagnostic study evaluating a decision-based algorithm for diagnosing pulmonary embolism (PE). Five radiologists, three board-certified general radiologists and two radiology trainees with 2 years' experience, participated in the study. RESULTS: According to the consensus reading, PE was present in 91 (31%) and absent in 194 (67%) patients, while in five patients (1.7%) the interpretations were regarded as equivocal. The per-patient agreement on the diagnosis of PE achieved by each of the four readers compared to the consensus reading was very good (kappa range 0.85-0.92), but peripheral emboli were missed in four to six patients by three of four observers. The agreement on the most proximal level of PE (per-proximal level) assessed by mean kappa value was 0.83 (kappa range 0.68-0.91) for the detection of proximal emboli, 0.61 for segmental emboli (kappa range 0.40-0.80), and 0.38 for emboli in the subsegmental vessels (kappa range 0.0-0.89). CONCLUSION: The overall agreement on the diagnosis of PE by MDCT for general radiologists and radiology trainees is very good, and we therefore believe that the initial management of patients with suspected PE could be based on the preliminary assessment performed by on-call radiologists with 2 years of experience.