Serological evidence of H9N2 avian influenza virus exposure among poultry workers from Fars province of Iran.

BACKGROUND: Since the 1990s, influenza A viruses of the H9N2 subtype have been causing infections in the poultry population around the globe. This influenza subtype is widely circulating in poultry and human cases of AI H9N2 have been sporadically reported in countries where this virus is endemic in domestic birds. The wide circulation of H9N2 viruses throughout Europe and Asia along with their ability to cause direct infection in mammals and humans, raises public health concerns. H9N2 AI was reported for the first time in Iran in 1998 and at present it is endemic in poultry. This study was carried out to evaluate the exposure to H9N2 AI viruses among poultry workers from the Fars province. METHODS: 100 poultry workers and 100 healthy individuals with no professional exposure to poultry took part in this study. Serum samples were tested for antibodies against two distinct H9N2 avian influenza viruses, which showed different phylogenetic clustering and important molecular differences, such as at the amino acid (aa) position 226 (Q/L) (H3 numbering), using haemagglutination inhibition (HI) and microneutralization (MN) assays. RESULTS: Results showed that 17 % of the poultry workers were positive for the A/chicken/Iran/10VIR/854-5/2008 virus in MN test and 12 % in HI test using the titer ≥40 as positive cut-off value. Only 2 % of the poultry workers were positive for the A/chicken/Iran/12VIR/9630/1998 virus. Seroprevalence of non exposed individuals for both H9N2 strains was below 3 % by both tests. Statistical analyses models showed that exposure to poultry significantly increases the risk of infection with H9N2 virus. CONCLUSIONS: The results have demonstrated that exposure to avian H9N2 viruses had occurred among poultry workers in the Fars province of Iran. Continuous surveillance programmes should be implemented to monitor the presence of avian influenza infections in humans and to evaluate their potential threat to poultry workers and public health.

Cumulus cell expansion and first polar body extrusion during in vitro oocyte maturation in relation to morphological and morphometric characteristics of the dromedary camel ovary.

The morphological and morphometric characteristics of the ovary are fundamental properties for in vitro oocyte maturation. Nuclear maturation, including first polar body (1PB) extrusion, cytoplasmic maturation and cumulus cell (CC) expansion are the criteria for in vitro maturation (IVM) of oocyte. This study was designed to determine the effect of morphological and morphometric features of the ovary on CC expansion and 1PB extrusion during IVM of oocyte in the adult female dromedary camel. The weight, volume and three dimensions of ovaries from slaughtered dromedary camels and oocytes inside zona diameter and zona pellucida thickness were measured. The follicles were classified in regard to the size and oocytes according to their ooplasm appearance and CC compactness. Aspirated cumulus oocyte complexes (COCs) were incubated for 48 hr (with a 6-hr interval) in Hams-F10, and CC expansion and 1PB extrusion were assessed. Significant differences were seen in the shape, weight, volume and three dimensions of the ovaries between ≤4-year-old and >4-year-old dromedary camel (p < .5). Approximately, 95.82% of follicles were 2-4 mm in diameter. The mean (±SD) of inside zona diameter of the oocyte and zona pellucida thickness was 132.22 ± 13.8 and 14.64 ± 2.24 µm, respectively, in >4-year-old dromedary camel. The CC expansion and 1PB extrusion were seen in 86% and 21.88% of COCs, respectively. Age and sexual conditions of dromedary camel influence the morphological and morphometric characteristics of the ovary. Most COCs retrieved from 2-6 mm follicles are cultivable. The most slaughterhouse-derived COCs retrieved from 2-6 mm follicles of non-pregnant dromedary camels are excellent and good and yielding a most favourable diameter to achieve the developmental competence for IVM in an optimal time of 24-30 hr; the optimal time for CC expansion is 24-30 hr in this species. However, the CC expansion is a prerequisite process, but not sufficient for IVM.

Pathogenesis of H9N2 virus in Chukar partridges.

Low-pathogenic avian influenza virus (H9N2) is circulating in the poultry industry of many countries in the Middle East and Asia, causing serious economic damage. In this study the clinical signs, antibody response, viral shedding and efficacy of oil emulsion vaccines in Chukar partridges were investigated until 9 days post inoculation (d.p.i.). Seventy-five Chukar partridges (Alectoris chukar) were divided randomly in three groups of challenged (Group C), vaccinated and challenged (Group VC) and control (non-vaccinated and non-challenged [Group NC]), 25 birds/group. Groups C and VC were inoculated with 0.4 ml allantoic fluid containing 10(7) median embryo infective dose/bird of A/Chicken/Iran/772/1998(H9N2) avian influenza virus. Clinical signs, antibody response, viral shedding and vaccine efficacy were evaluated and compared among these groups over 9 days. Clinical signs such as coughing and sneezing with depression and decreased feed and water consumption were observed in Group C. In Group VC only a slight decrease in food and water consumption was observed. Both Groups C and VC showed maximum antibody titre at 9 d.p.i. At 1 d.p.i. the virus was detected from all tissues in challenged group, but the virus was not detected from the spleen and caecal tonsil of Group VC. Group C showed the longest period of viral shedding in the trachea and kidney.

Frequency of sex chromosomal disomy in spermatozoa of normal and oligozoospermic Iranian patients and its effects on fertilisation and implantation rates after ICSI.

Chromosomal aneuploidy is a well-known phenomenon in human gametes including spermatozoa. Success rate of fertilisation and implantation in subfertile patients with male factor has always been shown to be very low. We tried to relate the possible impact of sex chromosomal aneuploidy in spermatozoa used for intracytoplasmic sperm injection (ICSI) on fertilisation and implantation rate. To evaluate the frequency of disomy for X and Y chromosomes in sperm samples retrieved from normal and oligozoospermic individuals, primed in situ labelling (PRINS) technique was used. Following ICSI, the rate of eight-cell embryos for each category was determined and followed up for successful implantation. Results showed a statistically significant higher frequency of disomy for all chromosomes under study in spermatozoa of oligozoospermic patients compared with normal men (P<0.01). The rate of eight-cells embryo formation was significantly lower than in normal group (P<0.01). The number of embryos transferred for both groups were nearly similar. Implantation rate for oligozoospermic patients was much lower than that of the normal group but was not significantly different (P>0.05). These results demonstrate that men especially with severe oligozoospermia have an elevated risk for chromosome abnormalities in their spermatozoa. These abnormalities might affect fertilisation and pre-embryo formation with less impact on implantation.

Construction of Meta-Thinking Educational Program Based on Mental-Brain Simulation (MTMBS) and Evaluating its Effectiveness on Executive Functions, Emotion Regulation, and Impulsivity in Children With ADHD: A Resting-State Functional MRI Study.

OBJECTIVE: The aim of present research was to make a Meta-Thinking educational program based on mental-brain simulation and to evaluate its effectiveness on executive functions, emotion regulation and impulsivity in children with ADHD. METHODS: The research method was Embedded Design: Embedded Experimental Model. The research sample included 32 children with ADHD who were randomly assigned to two experimental and control groups. The intervention was implemented for eight sessions of 1.5 hr for the experimental group, and fMRI images were taken from them, while the control group didn't receive any treatment. Finally, using semi-structured interviews, coherent information was collected from the parents of the experimental group about the changes made. Data were analyzed with SPSS-24, MAXQDA, fMRIprep, and FSL software. RESULTS: The Meta-Thinking Educational Program had effect on performance of ADHD children and suppressed brain regions related to DMN. CONCLUSION: The Implementation of this educational program plays a vital role in improving psychological problems of children with ADHD.

4K-memristor analog-grade passive crossbar circuit.

The superior density of passive analog-grade memristive crossbar circuits enables storing large neural network models directly on specialized neuromorphic chips to avoid costly off-chip communication. To ensure efficient use of such circuits in neuromorphic systems, memristor variations must be substantially lower than those of active memory devices. Here we report a 64 × 64 passive crossbar circuit with ~99% functional nonvolatile metal-oxide memristors. The fabrication technology is based on a foundry-compatible process with etch-down patterning and a low-temperature budget. The achieved <26% coefficient of variance in memristor switching voltages is sufficient for programming a 4K-pixel gray-scale pattern with a <4% relative tuning error on average. Analog properties are also successfully verified via experimental demonstration of a 64 × 10 vector-by-matrix multiplication with an average 1% relative conductance import accuracy to model the MNIST image classification by ex-situ trained single-layer perceptron, and modeling of a large-scale multilayer perceptron classifier based on more advanced conductance tuning algorithm.

Combinatorial optimization by weight annealing in memristive hopfield networks.

The increasing utility of specialized circuits and growing applications of optimization call for the development of efficient hardware accelerator for solving optimization problems. Hopfield neural network is a promising approach for solving combinatorial optimization problems due to the recent demonstrations of efficient mixed-signal implementation based on emerging non-volatile memory devices. Such mixed-signal accelerators also enable very efficient implementation of various annealing techniques, which are essential for finding optimal solutions. Here we propose a "weight annealing" approach, whose main idea is to ease convergence to the global minima by keeping the network close to its ground state. This is achieved by initially setting all synaptic weights to zero, thus ensuring a quick transition of the Hopfield network to its trivial global minima state and then gradually introducing weights during the annealing process. The extensive numerical simulations show that our approach leads to a better, on average, solutions for several representative combinatorial problems compared to prior Hopfield neural network solvers with chaotic or stochastic annealing. As a proof of concept, a 13-node graph partitioning problem and a 7-node maximum-weight independent set problem are solved experimentally using mixed-signal circuits based on, correspondingly, a 20 × 20 analog-grade TiO2 memristive crossbar and a 12 × 10 eFlash memory array.

Infection dynamics of highly pathogenic avian influenza and virulent avian paramyxovirus type 1 viruses in chickens, turkeys and ducks.

A range of virus doses were used to infect 3-week-old chickens, turkeys and ducks intranasally/intraocularly, and infection was confirmed by the detection of virus shedding from the buccal or cloacal route by analysis of swabs collected using real-time reverse transcriptase-polymerase chain reaction assays. The median infectious dose (ID(50)) and the median lethal dose (LD(50)) values for two highly pathogenic avian influenza (HPAI) viruses of H5N1 and H7N1 subtypes and one virulent Newcastle disease virus (NDV) were determined for each virus and host combination. For both HPAI viruses, turkeys were >100-fold more susceptible to infection than chickens, while both these hosts were >10-fold more susceptible to H5N1 virus than the H7N1 virus. All infected chickens and turkeys died. Ducks were also much more readily infected with the H5N1 virus (ID(50)< or =10(1) median embryo infective dose [EID(50)]) than the H7N1 virus (ID(50)=10(4.2) EID(50)). However, the most notable difference between the two viruses was their virulence for ducks, with a LD(50) of 10(3) EID(50) for the H5N1 virus, but no deaths in ducks being attributed to infection with H7N1 virus even at the highest dose (10(6) EID(50)). For both HPAI virus infections of ducks, the ID(50) was lower than the LD(50), indicating that infected birds were able to survive and thus excrete virus over a longer period than chickens and turkeys. The NDV strain used did not appear to establish infection in ducks even at the highest dose used (10(6) EID(50)). Some turkeys challenged with 10(6) EID(50), but not other doses, of NDV excreted virus for a number of days (ID(50)=10(4.6) EID(50)), but none died. In marked contrast, chickens were shown to be extremely susceptible to infection and all infected chickens died (ID(50)/LD(50)=10(1.9) EID(50)).

Molecular survey of avian circoviruses in some non-psittacine birds and detection of a novel canary circovirus in a pigeon.

BACKGROUND: Circoviruses are small, non-enveloped, single stranded DNA viruses. There is scarce information about these agents in non-psittacine birds. AIMS: It is attempted to detect and characterize circoviruses in non-psittacine birds. METHODS: Forty-five samples were collected from different non-psittacine species belonging to seven avian orders. A nested polymerase chain reaction (nested-PCR) for the detection of rep gene of circoviruses was applied. RESULTS: Two different types of circoviruses were detected in two pigeon samples (2/11, 18.2%). One of the detected circoviruses was placed in clade A next to a polish strain based on phylogenetic analysis. Interestingly, the other detected circovirus was closely related to canary circoviruses (CaCVs). CONCLUSION: In addition to the molecular diagnosis of a pigeon circovirus (PiCV), this is the first report of the detection of CaCv in a pigeon. The possible hypotheses of such circumstance are discussed.

Efficacy of Thermostable Newcastle Disease Virus Strain I-2 in Broiler Chickens Challenged with Highly Virulent Newcastle Virus.

Newcastle disease (ND) is a major threat to poultry industry production throughout developing countries. The Newcastle disease viruses (NDVs) infecting industrialized and indigenous poultry in Iran are velogenic strains and responsible for the frequent outbreaks of ND in poultry farms even in vaccinated flocks causing serious economic losses in the commercial and indigenous poultry. However, vaccination is the only way to protect against endemic ND, and the conventional vaccines are not heat stable and consequently require complex cold-chains to be transferred to users leading to not much resistance. The present study aimed to evaluate the efficacy of thermostable NDV strain I-2 in broiler chickens vaccinated via drinking water and coated on oiled wheat grain. The horizontal transmission of I-2 strain and transmission of disease from vaccinated to unvaccinated chickens were also evaluated in this study. The obtained results showed that both routes of administration, following primary and/or secondary dose, provoked the production of necessary antibody titer and adequate protective immunity in broiler chickens. Moreover, the horizontal transmission of I-2 strain from vaccinated to unvaccinated chickens housed together induced an antibody response and protected unvaccinated chickens against a local field isolate of a virulent strain of NDV (The intravenous pathogenicity index 2.46, mean death time 59 h). Nevertheless, all unvaccinated and Newcastle challenged broilers chickens against the NDV died in this study. It is noteworthy that the transmission of the virus from challenged broiler chickens was very low to induce clinical signs in susceptible chickens. The obtained results of this study revealed the efficacy of NDV strain I-2 coated on the oiled wheat and via drinking water as it protects broiler chickens from highly virulent NDV.

Molecular detection and characterization of beak and feather disease virus in psittacine birds in Tehran, Iran.

Beak and feather disease virus (BFDV), a member of genus circovirus, is a small, non-enveloped, single stranded DNA virus. Although BFDVs are among the most well studied circoviruses, there is little to no information about BFDVs in Iran. The aim of the present study was to detect and identify BFDV molecules from the birds referred to the avian clinic of The Faculty of Veterinary Medicine, Tehran University, Iran. A total of 55 DNA samples were extracted from birds from nine different species of the order psittaciformes. A robust conventional polymerase chain reaction (PCR) was applied to detect the rep gene of the virus. Ten out of 55 samples, from four different species, were tested positive for BFDVs in PCR (Melopsittacus undulates (4), Psittacula Krameri (3), Psittacus erithacus (2), Platycercus eximius (1)). Molecular identification of the detected BFDVs was performed based on their rep gene sequences. The phylogenetic analysis revealed that the Iranian BFDVs from this study were clustered into four genetically distinct clades belonging to different genetic subtypes of BFDVs (L1, N1, T1, and I4). Although the relation between the samples and their related subtypes in the tree are discussed, further studies are needed to elucidate the host specificity and incidence of the BFDVs from different genetic subtypes.

Implementation of multilayer perceptron network with highly uniform passive memristive crossbar circuits.

The progress in the field of neural computation hinges on the use of hardware more efficient than the conventional microprocessors. Recent works have shown that mixed-signal integrated memristive circuits, especially their passive (0T1R) variety, may increase the neuromorphic network performance dramatically, leaving far behind their digital counterparts. The major obstacle, however, is immature memristor technology so that only limited functionality has been reported. Here we demonstrate operation of one-hidden layer perceptron classifier entirely in the mixed-signal integrated hardware, comprised of two passive 20 × 20 metal-oxide memristive crossbar arrays, board-integrated with discrete conventional components. The demonstrated network, whose hardware complexity is almost 10× higher as compared to previously reported functional classifier circuits based on passive memristive crossbars, achieves classification fidelity within 3% of that obtained in simulations, when using ex-situ training. The successful demonstration was facilitated by improvements in fabrication technology of memristors, specifically by lowering variations in their I-V characteristics.

Spike-timing-dependent plasticity learning of coincidence detection with passively integrated memristive circuits.

Spiking neural networks, the most realistic artificial representation of biological nervous systems, are promising due to their inherent local training rules that enable low-overhead online learning, and energy-efficient information encoding. Their downside is more demanding functionality of the artificial synapses, notably including spike-timing-dependent plasticity, which makes their compact efficient hardware implementation challenging with conventional device technologies. Recent work showed that memristors are excellent candidates for artificial synapses, although reports of even simple neuromorphic systems are still very rare. In this study, we experimentally demonstrate coincidence detection using a spiking neural network, implemented with passively integrated metal-oxide memristive synapses connected to an analogue leaky-integrate-and-fire silicon neuron. By employing spike-timing-dependent plasticity learning, the network is able to robustly detect the coincidence by selectively increasing the synaptic efficacies corresponding to the synchronized inputs. Not surprisingly, our results indicate that device-to-device variation is the main challenge towards realization of more complex spiking networks.

Pathobiology of H9N2 avian influenza virus in Japanese quail (Coturnix coturnix japonica).

Clinical signs, serologic response, viral contents of the trachea and intestine, and histopathological and ultrastructural changes of the tracheal epithelium of Japanese quail experimentally infected with field isolate of H9N2 avian influenza were studied. Vaccinated and unvaccinated quail were inoculated with 10(6.3) 50% embryo infectious dose/bird of A/ chicken/Iran/SH-110/99 (H9N2) virus via nasal inoculation. Clinical signs such as depression, ruffled feathers, diarrhea, and nasal and eye discharges were observed 6 days postinfection (PI). No mortality was observed; however, there was reduction in feed and water consumption and egg production. However, the serologic response of vaccinated challenged and unvaccinated challenged birds was not significantly different. Unvaccinated challenged quail showed more severe histopathologic reaction in their lungs and trachea. Hyperemia, edema, infiltration of inflammatory cells, and deciliation and sloughing of the tracheal epithelium were observed. Ultrastructural study showed dilatation of endoplasmic reticulum and degeneration of Golgi apparatus and cilia of the tracheal lining cells of respiratory epithelium.

Efficacy of thermostable I-2 Newcastle disease vaccine compared to B1 commercial vaccine in broiler chicken.

Frequent vaccination failures have occurred in the broiler farms in Eurasian countries during Newcastle disease outbreaks. The disease is enzootic in many countries of the region, especially in southwest Asia. I-2 vaccine has been used successfully in village chickens in many Asian and African countries. Our preliminary study showed good efficacy of the vaccine in broiler chickens. Therefore the current experimental study was conducted to compare viral shedding period of heat resistance I-2 vaccine with B1 commercial vaccine following challenge with Herts'33. For this purpose three hundred commercial broilers were randomly allocated into four groups; 1) Thermostable I-2 vaccine, 2) Hitchner B1 vaccine, 3) Challenge group with no vaccine, and 4) Negative control group. Experimental chicks were vaccinated on days 19 and 26 by the eye drop route and then the birds were challenged via intra ocular route on day 40 with a suspension containing 106 EID50/ml challenge virus. Experimental chickens were monitored by collecting buccal and cloacal swabs at different times. Collected swabs were submitted to PCR test. The results showed that vaccination can protect the birds against mortality and also decrease virus shedding; also there was not a significant difference between vaccination with I-2 and B1 vaccines.

Chronological and ultrastructural changes in camel (Camelus dromedarius) oocytes during in vitro maturation.

Cumulus-oocyte complexes (COCs) were collected from non-pregnant camels at a local slaughterhouse by aspiration from antral follicles (2-6 mm). In Experiment I, camel COCs (n=304) were matured in vitro in Hams-F10, fixed at different time intervals (6, 12, 18, 24, 30, 36, 42, or 48 h) and stained with 1% aceto-orcein to assess nuclear changes in culture. A majority of the oocytes (81.5%) underwent germinal vesicle break down (GVBD) between 6 and 12h. Forty-eight percent of the oocytes were observed at the metaphase I (M I) stage by 18 h culture. The percentage of matured oocytes (M II stage) at 30 and 42 h were 66.5 and 71% respectively, which were significantly (p<0.05) different to that observed at 24 h (42.5%). In Experiment II, after different periods of culture (12, 24, 36, or 48 h), the COCs (n=26) were processed for transmission electron microscopy. Expansion of both the cumulus and corona radiate cells occurred between 12 and 24 h in the majority of oocytes concomitant with enlargement of the cumulus cell process endings (CCPEs) in the developed perivitelline space. After 12 h of culture disruption of the junctions between CCPEs and the oolemma was observed together with and breakdown of the GV. For 24-36 h of culture cortical granules had spread and aligned along the oolemma. Signs of degeneration in the cytoplasmic organelles of the oocytes were also observed from less than 36 h. After 48 h of culture, larger vesicles and lipid droplets had appeared in the central part of the oocytes and showed uneven distribution throughout the ooplasm. Predominantly non-penetrating CCPEs were also observed in four oocytes by 48 h. In conclusion, based on both light and electron microscopic evaluations, the optimal culture time for the development of competent Camelus dromedarius oocytes in vitro appears to be 30 h using Hams-F10 medium.

Avian influenza (H9N2) outbreak in Iran.

An epidemic of avian influenza (AI) (H9N2) occurred in broiler chicken farms in Iran during 1998-01. Mortality between 20% and 60% was commonly observed on the affected farms. Mixed infections of the influenza virus with other respiratory pathogens, particularly infectious bronchitis virus and Mycoplasma gallisepticum, were thought to be responsible for such high mortality, which resulted in great economic losses. Clinical signs included swelling of the periorbital tissues and sinuses, typical respiratory discharge, and severe respiratory distress. Gross lesions included extensive hyperemia of the respiratory system followed by exudation and cast formation in the tracheal biforcation extending into the secondary bronchi. Light microscopy lesions were characterized by severe necrotizing tracheatis. Serological examination using H9N2 AI viral antigen produced inconsistent results. Ultrastructural findings showed typical viral replication through budding processes on cell membranes of the tracheal epithelium.

Can dogs carry the global pandemic candidate avian influenza virus H9N2?

BACKGROUND: H9N2 avian influenza virus (AIV) is one of the most widely circulating viruses in Eurasia. Recent studies have shown that the molecular recombination of H9N2 and H1N1 could pose a pandemic threat. Mammals that are susceptible to subtype H9N2 may contribute to the spread of the virus. OBJECTIVE: To determine the susceptibility of 1-year-old dogs to H9N2 AIV. PROCEDURE: H9N2 AIV infection was experimentally reproduced in 1-year-old dogs. The animals were intranasally inoculated with a titre of 10(7.5) (50% egg infective dose) of H9N2 AIV isolated from a broiler farm during an outbreak. The animals in the contact group were exposed to contaminated surfaces. RESULTS: Clinical signs including sneezing, coughing and nasal discharge were observed in the inoculated and contact groups. The virus was detected in nasal swab, faecal and buffy coat samples of dogs in both the inoculated and contact groups and both groups developed antibody titres against AIV H9N2 subtype. CONCLUSION: H9N2 AIV isolated from outbreaks in a broiler farm can easily infect dogs and infected animals shed the virus. Because many Asian countries are facing frequent outbreaks of H9N2 infection in the poultry industry, dogs could be a potentially important source of virus transmission within and between poultry farms.