The burden of lower respiratory infections and their underlying etiologies in the Middle East and North Africa region, 1990-2019: results from the Global Burden of Disease Study 2019.

BACKGROUND: Lower respiratory infections (LRIs) cause substantial mortality and morbidity. The present study reported and analysed the burden of LRIs in the Middle East and North Africa (MENA) region between 1990 and 2019, by age, sex, etiology, and socio-demographic index (SDI). METHODS: The data used in this study were sourced from the Global Burden of Disease (GBD) study 2019. The annual incidence, deaths, and disability-adjusted life-years (DALYs) due to LRIs were presented as counts and age-standardised rates per 100,000 population, along with their 95% uncertainty intervals (UIs). The average annual percent changes (AAPC) in the age-standardised incidence, death and DALYs rates were calculated using Joinpoint software and correlations (Pearson's correlation coefficient) between the AAPCs and SDIs were calculated using Stata software. RESULTS: In 2019, there were 34.1 million (95% UI 31.7-36.8) incident cases of LRIs in MENA, with an age-standardised rate of 6510.2 (95% UI 6063.6-6997.8) per 100,000 population. The number of regional DALYs was 4.7 million (95% UI 3.9-5.4), with an age-standardised rate of 888.5 (95% UI 761.1-1019.9) per 100,000 population, which has decreased since 1990. Furthermore, Egypt [8150.8 (95% UI 7535.8-8783.5)] and Afghanistan [61.9 (95% UI 52.1-72.6)] had the highest age-standardised incidence and death rates, respectively. In 2019, the regional incidence and DALY rates were highest in the 1-4 age group, in both females and males. In terms of deaths, pneumococcus and H. influenza type B were the most and least common types of LRIs, respectively. From 1990 to 2019, the burden of LRIs generally decreased with increasing SDI. There were significant positive correlations between SDI and the AAPCs for the age-standardised incidence, death and DALY rates (p < 0.05). Over the 1990-2019 period, the regional incidence, deaths and DALYs attributable to LRIs decreased with AAPCs of - 1.19% (- 1.25 to - 1.13), - 2.47% (- 2.65 to - 2.28) and - 4.21% (- 4.43 to - 3.99), respectively. CONCLUSIONS: The LRI-associated burden in the MENA region decreased between 1990 and 2019. SDI had a significant positive correlation with the AAPC and pneumococcus was the most common underlying cause of LRIs. Afghanistan, Yemen and Egypt had the largest burdens in 2019. Further studies are needed to investigate the effectiveness of healthcare interventions and programs to control LRIs and their risk factors.

Effects of Various Anticoagulants on Biochemistry Analytes From Domestic Pigeons (Columba livia domestica).

The objective of this study was to compare the effects of anticoagulant and no anticoagulant on routine biochemical analytes in domestic pigeons (Columba livia domestica). Blood samples were obtained from 8 clinically healthy pigeons. The sample obtained from each bird was divided into 4 blood collection tubes containing either ethylenediaminetetraacetic acid (EDTA), lithium heparin, sodium citrate, or no anticoagulant. The concentrations of creatinine, uric acid, triglyceride, total cholesterol, glucose, phosphorus, calcium, magnesium, total protein, albumin, and iron, and the activities of alkaline phosphatase (ALP), alanine aminotransferase (ALT), and aspartate aminotransferase (AST), were measured in blood from each of the blood collection tubes. The values of the measured parameters, with the exception of iron, were significantly lower in the citrated plasma samples compared with the serum samples, even after correcting for dilution. In the lithium heparin plasma samples, significant decreases in albumin, triglyceride, calcium, total cholesterol, and ALP, and a significant increase in iron, were observed compared with the values in the serum samples. The concentrations of total protein, creatinine, glucose, total cholesterol, triglyceride, magnesium, calcium, and phosphorus, as well as the activities for AST and ALP, were significantly lower in the EDTA plasma samples compared with the serum samples. In conclusion, the anticoagulants had significant effects on most of the measured parameters compared with serum. The findings of the present study suggest that a lithium heparin sample is the most appropriate plasma sample for the measurement of blood biochemical parameters in the domestic pigeon.

Radiographic Reference Intervals for Cardiac Size in Common Mynahs (Acridotheres tristis).

Cardiovascular diseases in the common mynah (Acridotheres tristis) often go undiagnosed due to limitations with the physical examination and common concurrent illnesses. The aim of this study was to establish accurate reference intervals for cardiac size based on radiographs for this species. All birds were considered healthy based on physical examination. High-quality ventrodorsal radiographic images were obtained from 34 healthy common mynah birds. The cardiac width (CW), thoracic width, distance between third and fourth ribs, synsacral width, coracoid width, and distance between clavicular bones were evaluated on the ventrodorsal radiographic images. Additionally, the ratio between the CW and these anatomical indices were measured. In order to evaluate the correlation between CW and the radiographic indices, a linear regression model was used. The mean ± SD for CW in the 34 birds was 16.1 ± 0.9 mm (95% confidence interval: 14.1-18 mm). Significant moderate correlation was found for CW with thoracic width (R2 = 0.37; P < 0.001), coracoid width (R2 = 0.25; P = 0.002), and the distance between the third and fourth ribs (R2 = 0.34; P < 0.001). The radiographic reference intervals calculated in this study can be used to evaluate cardiac size in the common mynah bird.

Comparison of iron chelators used alone or in combination with phlebotomy in common mynahs (Acridotheres tristis) with experimental iron storage disease.

To compare the therapeutic effects of iron chelators used alone or in combination with phlebotomy on iron storage disease (ISD), 66 healthy common mynahs (Acridotheres tristis) were fed an iron-loading diet (3000 ppm) for 30 days. After confirmation of ISD, the birds were randomly divided into four treatment groups; DFO: deferoxamine (100 mg/kg SC q24 h), DFP: deferiprone (oral, 75 mg/kg), DFO + F: deferoxamine (100 mg/kg SC q24 h) with phlebotomy, and DFP + F: deferiprone (oral, 75 mg/kg) with phlebotomy. In phlebotomy-treated groups, blood sampling (1% BW) was performed weekly. At 1 and 2 months after treatments, seven birds from each group were euthanized and liver iron, copper, and zinc were analysed by ICP-OES assay. After 1 month, in all treatments, the liver amount of iron, copper, and zinc was reduced (P < 0.05) and there was no significant difference between groups. In the second month, the amount of liver iron, copper, and zinc decreased more in all groups, but this change was insignificant except in the DFP + F group (P < 0.05). These results suggest that all therapeutic protocols after 1 month effectively reduce the liver iron and there is no need to continue treatment. Otherwise, it may lead to iron deficiency, especially in birds treated with DFP + P. Since deferiprone, as an inexpensive oral chelator, effectively reduces liver iron levels without causing stress in the birds, it can be recommended as a more appropriate method for the treatment of mynahs with ISD. However, further clinical studies are needed to define the most effective treatment.RESEARCH HIGHLIGHTS Deferiprone is an optimized method for treating iron storage disease.The essential metals homeostasis is impaired in iron storage disease.

Protein Electrophoresis of Serum and Heparinized Plasma in the Common Mynah (Acridotheres tristis).

Although serum protein electrophoresis is a diagnostic tool available through many veterinary laboratories, there currently are no reference intervals for protein fractions in healthy common mynahs (Acridotheres tristis). Therefore, electrophoretic patterns of proteins in serum and heparinized plasma of the common mynah were evaluated. Blood specimens were collected from 55 healthy adult common mynahs of unknown age (26 males and 29 females). The serum total protein and protein fractions were measured using the biuret method followed by cellulose acetate electrophoresis (CAE). The serum level of albumin was compared with bromocresol green (BCG) dye-binding and CAE methods. Four protein fractions, including albumin and α, ß, and γ globulins, were recorded in the electrophoretogram of serum specimens. Sex appeared to have no significant effect on the measured parameters. The serum BCG albumin fraction was significantly higher than the CAE albumin fraction (P = .01). Also, the comparison of total protein and protein fractions in serum and plasma specimens of 25 of the 55 birds sampled showed that total protein (Cohen index d = 0.66, P = .03), gamma globulin (d = 1.13, P = .00), and total globulin (d = 0.67, P = .00) in plasma samples were significantly higher than those in serum samples. The results of this study provide the specific reference intervals for total protein and protein fractions in common mynahs, which are essential for proper interpretation of laboratory results and also revealed that the albumin measurement by the BCG method yields unreliable results in common mynahs.

Osteofibrous Dysplasia in a Cockatiel (Nymphicus hollandicus).

Osteofibrous dysplasia is a rare and benign nonneoplastic condition of unknown etiology in humans and mammals. An adult female cockatiel (Nymphicus hollandicus) was presented with the following problems: a soft tissue mass on the left frontal area of the head that extended over the left eye, anorexia, lethargy, and emaciation. Computed tomographic imaging and ultrasonography revealed a soft tissue opacity in the left cranial area of the head with multifocal heterogeneous opacities and foci of mineralization. Cytological, histopathological, and immunohistochemistry examinations were performed on biopsy samples from tissue collected from the mass. Following surgical removal of the cranial mass the patient was treated with a nonsteroidal anti-inflammatory drug and antibiotic medication. The surgical site healed with no complications and through follow-up examinations there were no signs of reoccurrence of the abnormal tissue for 2 months. Through cytological and histopathological examination and immunohistochemistry testing of the submitted tissue, a diagnosis of osteofibrous dysplasia was determined.

Characterization of Iranian canarypox and pigeonpox virus strains.

Avipoxviruses (APVs) are large DNA viruses that are detected widely in many species of birds. Little information is available regarding genetic variations in these host-specific viruses. In the present study, nine canarypox virus and five pigeonpox virus isolates were collected from northeastern Iran and isolated via the chorioallantoic membrane of chicken embryos. Further investigations were conducted using analysis of virus growth in chicken embryo fibroblasts, histopathology, electron microscopy, and molecular techniques such as polymerase chain reaction (PCR) combined with sequencing and phylogenetic analysis to investigate variations in the highly conserved P4b gene of poxviruses. Virus replication and pock lesions were evident, and microscopic examination revealed eosinophilic intracytoplasmic inclusion bodies and biconcave enveloped virus particles with randomly arranged surface filaments, which are characteristic features of poxviruses. PCR results confirmed the presence of an APV-specific 578-bp fragment in all of the samples. Sequence analysis and phylogenetic analysis of 578-bp P4b fragments of eight isolates confirmed that our canary and pigeon isolates clustered with previously reported isolates. The similarity between the nucleotide sequences of most of our isolates and those isolated previously in other countries could be due to the high degree of conservation of these fragments. However, the FZRC6V isolate from a canary in this study did not have a canarypox virus origin according to the sequence analysis, and might have originated from cross-infection with different strains of avipoxviruses.

Isolation and toxin gene detection of Clostridium (Clostridioides) difficile from traditional and commercial quail farms and packed quail meat for market supply - Short communication.

Clostridium (Clostridioides) difficile is a Gram-positive anaerobic rod-shaped bacterium and the main cause of nosocomial diarrhoea in humans. In recent years, the transmission of C. difficile from environmental reservoirs (e.g. food) to humans has become a major focus of research. The aim of this study was to investigate the prevalence and corresponding toxin genes of C. difficile in faecal samples and meat of quails. Thirty samples of packed quail meat in Mashhad, Iran and 500 faecal samples (pooled to n = 5) were collected on quail farms in the Northeastern Khorasan region for further investigation. Of 100 pooled quail faecal samples 10% showed cultural growth of C. difficile. In meat samples two out of 30 specimens (7%) showed cultural growth. In six of ten isolates from faecal samples toxin genes (tcdB and tcdA) were present, while four isolates harboured no toxin genes. However, in meat isolates no toxin genes were present. Mutations in the tcdC gene were not detected, indicating that 'hypervirulent' strains such as RT027 and RT078 were not present. The data suggest that quail and quail products might hold a potential for the spread of C. difficile.

High Prevalence of Cryptococcus neoformans and Isolation of Other Opportunistic Fungi From Pigeon (Columba livia) Droppings in Northeast Iran.

Cryptococcus neoformans is an opportunistic human pathogen that causes cryptococcosis, a life-threatening infection that usually manifests as meningoencephalitis in immunocompromised patients. Pigeon (Columba livia) droppings can spread pathogenic yeasts and mold fungi, such as C neoformans, in the environment. The objective of this study was to isolate C neoformans and other opportunistic fungi from feral pigeon droppings. One hundred twenty samples of feral pigeon droppings were suspended 1:10 in saline solution and then cultured. The fungi were identified by standard mycological techniques. Fungal contamination was detected in all examined samples. Yeast and mold fungi were isolated from 114 samples (95%) and 103 samples (85.8%), respectively, out of 120 samples. The highest frequency of yeast and mold fungi isolated from collected samples was C neoformans 77.5% and Rhizopus species 38.3%, respectively. Several types of fungi exist in pigeon droppings that can be spread in the environment and transmitted to children and elderly, as well as immunocompromised patients who are at increased risk of contracting opportunistic diseases.

Plasma Biochemical and Lipid Panel Reference Intervals in Common Mynahs (Acridotheres tristis).

Mynahs are popular pet and aviary birds. Captive mynahs are prone to metabolic disorders, such as obesity, hepatic lipidosis, and atherosclerosis, and determination of a plasma biochemical reference interval is essential for diagnosis, treatment, and monitoring of these disorders. To establish reference intervals for the lipoprotein panel and plasma activity of liver enzymes in common mynahs (Acridotheres tristis), glucose, cholesterol, triglycerides, high and low density lipoprotein concentrations, and activities of aspartate aminotransferase, alkaline phosphatase, gamma-glutamyl transferase, and lactate dehydrogenase were measured in 56 adult healthy individuals. The data were grouped according to sex and body weight. For all variables, no significant differences were found between male and female mynahs, and body weight had no effect on lipoprotein fractions. Our results provide a reliable reference interval for some blood biochemical values in healthy common mynahs that could be useful for diagnosing and monitoring disease problems.

Recombinant production of a chimeric antimicrobial peptide in E. coli and assessment of its activity against some avian clinically isolated pathogens.

Over the last decades, poultry industry faced to the rapid emergence of multidrug-resistant bacteria as a global concern. Antimicrobial peptide (AMPs) known as potential antibiotic alternative and were considered as a new antimicrobial agent. Current methods of production and purification of AMPs have several limitations such as: costly, time-consuming and killing the producing host cells in recombinant form. In the present study, a chimeric peptide derived from camel lactoferrin was produced in Escherichia coli periplasmic space using a pET-based expression system and its antibacterial activity was determined on some avian pathogens in vitro. A carboxy-terminal polyhistidine tag was used for purification by Ni2+ affinity chromatography with an average yield of 0.42 g/L. The His-tagged chimeric peptide showed different range of antimicrobial activity against clinically isolated avian pathogens with low chicken blood hemolysis activity and high serum stability. Overall, the results of this investigation showed the recombinant chimeric peptide was successfully expressed in pET-based expression system and could be considered as a proper alternative for some currently used antibiotics in poultry industry and drugs veterinary medicine.

Evaluation of Cardiac Size in the Common Kestrel ( Falco tinnunculus ) Based on Radiographic Measurements.

The common kestrel ( Falco tinnunculus ) is a common raptor found worldwide. To establish reference intervals for cardiac size by radiography, 14 adult common kestrels were evaluated clinically and with radiographs (lateral and ventrodorsal views). The cardiac width, thoracic width, distance between third and fourth ribs, synsacrum width, coracoid width, and distance between clavicle bones were measured on ventrodorsal radiographs. The ratio between cardiac width and the other 5 indices was calculated. Results showed a significant, positive correlation between the cardiac width and thoracic width and between cardiac width and coracoid width. The values and ratios of this study can be used as reference intervals for cardiac size evaluated radiographically in the common kestrel for diagnosis of the cardiomegaly in this bird.

Radiographic reference limits for cardiac width of budgerigars (Melopsittacus undulatus).

Primary and secondary cardiovascular diseases are not uncommon in birds. Although radiologic standards for heart width have been developed for mammals, they are still not available for many avian species. The purpose of this study was to establish normal reference values for cardiac size in budgerigar (Melopsittacus undulatus), one of the most popular pet bird species all over the world. After clinical and radiographic (lateral and ventrodorsal views) evaluations, 27 adult, clinically healthy budgerigars (10 females and 17 males) were included in this study. High-quality ventrodorsal and lateral radiographic projections were obtained. The cardiac and thoracic width, distance between third and fourth ribs, synsacrum width, coracoid width, and the distance between clavicle bones were measured on ventrodorsal radiographs. The ratio between cardiac width and other mentioned indices was calculated. Correlation of each anatomical index with the cardiac width was evaluated by linear regression model. Sex and weight were included in all models. Mean + SD of cardiac width was 10.8 +/- 0.6 mm, with lower and upper limits of 9.5 and 12.0 mm. The results showed a significant correlation between the cardiac width and the thoracic width (R2 = 0.28; P = 0.005). There were no significant associations between weight, sex, and the heart width. The values and ratios obtained in this study can be used as a reference of normal cardiac size of budgerigar in radiology for detection of cardiomegaly in this bird.

Comparison of a Point-of-Care Glucometer and a Laboratory Autoanalyzer for Measurement of Blood Glucose Concentrations in Domestic Pigeons ( Columba livia domestica).

Biochemical analysis is necessary for diagnosis and monitoring of diseases in birds; however, the small volume of blood that can be safely obtained from small avian species often limits laboratory diagnostic testing. Consequently, a suitable methodology requiring only a small volume of blood must be used. This study was designed to compare blood glucose concentrations in domestic pigeons ( Columba livia domestica) as measured by a commercial, handheld, human glucometer and a standard autoanalyzer. During the first phase of the study, whole blood samples obtained from 30 domestic pigeons were used to measure the blood glucose concentration with a glucometer, the packed cell volume (PCV), and the total erythrocyte count (nRBC). Plasma separated from the each sample was then used to obtain the plasma glucose concentration with the autoanalyzer. During the second phase of the study, 30 pigeons were assigned to 2 equal groups (n = 15). Hypoglycemia or hyperglycemia was induced in each group by intravenous injection of insulin or glucose, respectively. Blood was collected and processed, and glucose concentrations, PCV, and nRBC were measured as previously described. Linear-regression models demonstrated a significant relationship between results measured by the glucometer and autoanalyzer results from normoglycemic (correlation coefficient [R] = 0.43, P = .02), hypoglycemic (R = 0.95; P < .001), and hyperglycemic (R = 0.81; P < .001) birds. The results of this study suggest that we can predict the real blood-glucose concentration of pigeons by using results obtained by a glucometer.

A Review on cLF36, a Novel Recombinant Antimicrobial Peptide-Derived Camel Lactoferrin.

Lactoferrin is an antimicrobial peptide (AMP) playing a pivotal role in numerous biological processes. The primary antimicrobial efficacy of lactoferrin is associated with its N-terminal end, which contains various peptides, such as lactoferricin and lactoferrampin. In this context, our research team has developed a refined chimeric 42-mer peptide known as cLF36 over the past few years. This peptide encompasses the complete amino acid sequence of camel lactoferrampin and partial amino acid sequence of lactoferricin. The peptide's activity against human, avian, and plant bacterial pathogens has been assessed using different biological platforms, including prokaryotic (P170 and pET) and eukaryotic (HEK293) expression systems. The peptide positively influenced the growth performance and intestinal morphology of chickens challenged with pathogen bacteria. Computational methods and in vitro studies showed the peptide's antiviral effects against hepatitis C virus, influenza virus, and rotavirus. The chimeric peptide exhibited higher activity against certain tumor cell lines compared to normal cells, which may be attributed to the peptide's interaction with negatively charged glycosaminoglycans on the surface of tumor cells. Importantly, this peptide exhibited no toxicity against host cells and demonstrated remarkable thermal and protease stability in serum. In conclusion, while our investigations suggest that the chimeric peptide, cLF36, may offer potential as a candidate or complementary option to some available antibiotics, antiviral agents, and chemical pesticides, significant uncertainties remain regarding its cost-effectiveness, as well as its pharmacodynamic and pharmacokinetic characteristics, which require further elucidation.

Molecular detection and analysis of beak and feather disease viruses in Iran.

The beak and feather disease virus (BFDV) is one of the few pathogens capable of causing extinction of psittacines. To determine the prevalence and the nature of BFDV mutation, this study investigated the presence of the BFDV among 1,095 individual birds of the 17 psittacine species in Iran followed by analyzing the DNA sequences of seven replication-associated protein (rep) and 10 capsid (cap) genomes of the virus. The BFDV was found to be the foremost pathogen among more than 12 psittacine species, and phylogenetic analysis showed that the BFDV GenBank-published sequences from Poland, Saudi Arabia, South Africa, Taiwan, and Thailand were most similar to those of this study. Evolutionary analysis concluded that arginine, leucine, and glycine were the amino acids frequently involved in the least-conserved substitution patterns of BFDV, and conversely, methionine, glutamine, and tryptophan were the amino acids that exhibited ultra-high conservation through the substitution patterns. The high substitution rate of arginine to lysine and glycine to serine also made greater contribution to the BFDV gene mutation. The relative synonymous codon usage between two genes revealed that the cap genome encoded proteins frequently used fewer codons, while the rep genome encoded proteins used more codons only at moderate frequency, explaining the broader divergence of the cap compared to the rep sequence. The data analysis also introduced a new variant of BFDV that exists in the rep and cap sequences of budgerigars. While the existence of more new variants was suspected, more solid evidence is required to substantiate this suspicion.

Intestinal changes and immune responses during Clostridium perfringens-induced necrotic enteritis in broiler chickens.

Clostridium perfringens-induced necrotic enteritis (NE) is an economically important disease of broiler chickens. The present study evaluated the effect of C. perfringens on the intestinal histomorphometry, enteric microbial colonization, and host immune responses using 3 experimental NE reproduction methods. The experimental groups consisted of 1) unchallenged Control diet (corn-soybean meal), 2) Control diet + Eimera inoculation at d 11 followed by C. perfringens challenge at d 15 (ECp), 3) Wheat-based diet + C. perfringens challenge (WCp), and 4) Wheat-based diet + Eimeria inoculation followed by C. perfringens challenge (WECp). The results showed that chickens receiving ECp and WECp had reduced (P < 0.05) bird performance coupled with enteric gross lesions and epithelial damage at d 17 and 24 of age compared to unchallenged control birds. These ECp and WECp administered birds also had increased (P < 0.05) ileal colonization by clostridia and E. coli at d 17 and 24, while the resident Lactobacillus counts were reduced (P < 0.05) at d 24 of age. Furthermore, at d 24, jejunal transcription of IL-6, IL-10, annexin-A1 and IL-2 genes was upregulated (P < 0.05) in the ECp group, whereas the transcription of TNF receptor associated factor (TRAF)-3 gene was increased (P < 0.05) in WECp treated birds when compared to unchallenged control group. Additionally, stimulation of chicken splenocytes and cecal tonsilocytes with virulent C. perfringens bacilli or their secretory proteins resulted in a higher (P < 0.05) frequency of T cells and their upregulation of MHC-II molecule, as determined by flow cytometry. These findings suggest that C. perfringens, while inducing epithelial damage and changes in microbiota, can also trigger host immune responses. Furthermore, NE reproduction methods using coccidia with or without the wheat-based dietary predisposition seem to facilitate an optimal NE reproduction in broiler chickens and thus, may provide better avenues for future C. perfringens research.

Effects of nano-manganese on humoral immune response and oxidative stress in broilers.

The objective of the present study was to evaluate the alterations in selected indicators of immune responses and oxidative stress of broilers fed with nano-manganese. One hundred-sixty 1-day-old broiler chicks were randomly assigned into four groups with three replicates. Birds were fed the same basal diet supplemented with nano-manganese oxide, as 0.00 (control group), 50.00, 100, or 150 mg kg-1 of diet. The birds were vaccinated against avian influenza (AI), Newcastle disease (ND), infectious bronchitis (IB) and infectious bursal disease (IBD) as the standard vaccination schedule. Blood sample was taken from the brachial vein of birds on 42th day. A significant decrease in antibody titer against sheep RBC was revealed in the nano-manganese 100 and 150 groups compared to the control group. In addition, the antibody titers against IB and ND were significantly lower in the all nano-manganese groups compared to the control group. No significant difference was observed for the antibody titer against AI and oxidative stress indices among the experimental groups. The findings in the present study suggested that nano-manganese at 50.00, 100 and 150 mg kg-1 levels might suppress humoral immune response in broilers which should be taken into consideration in supplementation.

Mesoporous silica nanoparticles-based formulations of a chimeric proteinous vaccine candidate against necrotic enteritis disease.

To develop a nanoparticle-based vaccine against necrotic enteritis, a chimeric antigen (rNA) consisting of the main antigens of Clostridium perfringens, NetB, and Alpha toxin, was prepared. Then, the rNA molecules were loaded onto the functionalized mesoporous silica nanoparticles (MSNPs) using physical adsorption or covalent conjugation methods. The characterization of synthesized nanoparticles was performed by scanning electron microscopy, dynamic light scattering, zeta potential measurement, Fourier transform infrared spectroscopy, and thermogravimetry techniques. The results revealed that the spherical nanoparticles with an average diameter of 90 ±â€¯12 nm and suitable surface chemistries are prepared. MSNPs-based formulations did not show any significant toxicity on the chicken embryo fibroblast cells. The results of the challenge experiments using subcutaneous or oral administration of the as-prepared formulations in the animal model showed that the as-prepared nanosystems, similar to those formulated with a commercial adjuvant (Montanide), present stronger humoral immune responses as compared to that of the free proteins. It was also indicated that the best protection is obtained in groups vaccinated with MSNPs-based nanovaccine, especially those who orally received covalently conjugated nanovaccine candidates. These results recommend that the MSNPs-based formulated chimeric proteinous vaccine candidates can be considered as an effective immunizing system for the oral vaccination of poultry against gastrointestinal infectious diseases.

Serologic and Molecular Evidence of Widespread Infection of Avian Hepatitis E Virus in Poultry Farms of Iran.

Hepatitis-splenomegaly syndrome is caused by avian hepatitis E virus (aHEV), a nonenveloped, single-stranded RNA virus. The economic importance of this disease in the poultry industry is due to the decline in egg production (10%-40%) and the rise in mortality (1%-4%). In the present study, 1540 serum samples from 33 broiler breeder flocks were analyzed by an enzyme-linked immunosorbent assay for the presence of an anti-aHEV antibody. In addition, a diagnostic nested reverse transcriptase-PCR was done on all farm samples. In the serologic study, 66.7% (22/33) of the flocks and 28.5% (439/1540) of the chickens were positive. The molecular study showed that three farms were positive, and PCR products were observed for the conserved regions of the aHEV helicase and capsid virus genes as 386 bp and 242 bp, respectively. It should be noted that clinical and pathologic symptoms including decreased egg production, enlarged livers and spleens, and a slight rise in mortality rate were observed in eight farms. To our knowledge, this is the first documented study on the aHEV identification and its antibody detection in broiler breeder farms in Iran.

Evidencia serológica y molecular de una infección diseminada del virus de la hepatitis E aviar en granjas avícolas en Irán. El síndrome de hepatitis-esplenomegalia es causado por el virus de la hepatitis E aviar (aHEV), un virus de ARN de cadena simple sin envoltura. La importancia económica de esta enfermedad en la industria avícola se debe a la disminución en la producción de huevo (10%-40%) y al aumento de la mortalidad (1%-4%). En el presente estudio, se analizaron 1540 muestras de suero de 33 parvadas de reproductores pesados mediante un ensayo de immunoabsorción con enzimas ligadas para determinar la presencia de anticuerpos contra el virus de la hepatitis E aviar. Además, se realizó un método de transcripción reversa y PCR anidado de diagnóstico en todas las muestras de la granja. En el estudio serológico, el 66.7% (22/33) de las parvadas y el 28.5% (439/1540) de los pollos fueron positivos. El estudio molecular mostró que tres granjas fueron positivas, y se observaron productos de PCR para las regiones conservadas de los genes del virus de la cápside y de la helicasa del virus de la hepatitis E aviar con tamaños de 386 pb y 242 pares de bases, respectivamente. Cabe señalar que en ocho granjas se observaron signos clínicos y patológicos como disminución de la producción de huevos, agrandamiento del hígado y del bazo y un ligero aumento en la tasa de mortalidad. Hasta donde se conoce, este es el primer estudio documentado sobre la identificación del virus de la hepatitis E aviar y la detección de anticuerpos en granjas de pollos de engorde en Irán.