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.

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.

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.

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.

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.

Immunization with oral and parenteral subunit chimeric vaccine candidate confers protection against Necrotic Enteritis in chickens.

Following the ban on the use of in-feed antimicrobials, necrotic enteritis (NE) NE is the most important clostridial disease. Vaccination has been considered as a possible approach to prevent NE. Our previous study showed that a chimeric protein product consisting of antigenic epitopes of NetB, Alpha-toxin and Zinc metallopeptidase (Zmp) triggered immune response against C. perfringens. In the current study we optimized the chimeric gene and constructed a fusion protein containing NetB, Alpha-toxin and Metallopeptidase (NAM) for expressing in tobacco plant to use as an edible vaccine for immunizing the chicken against NE. Simultaneously, we expressed and purified a His-tagged recombinant version of the NAM (rNAM) expressed in E. coli BL21 for subcutaneous immunization of chickens. Immunized birds produced strong humoral immune responses against both edible plant-based and parenteral purified rNAM. The responses were determined by the mean titer of antibody in blood samples to be around 9000 and 32,000, for edible and injected rNAM, respectively. Birds immunized subcutaneously showed the most striking responses. However the edible vaccine provided a more long lasting IgY response 14 days after the third vaccination compared to the injected birds. Chickens immunized with either lyophilized leaves expressing rNAM or purified rNAM, subsequently were subjected to the challenge with a virulent C. perfringens strain using an NE disease model. Our results showed that birds immunized both parenterally and orally with recombinant chimeric vaccine were significantly protected against the severity of lesion in the intestinal tract, but the protection provided with the injectable form of the antigen was greater than that of the oral form. Further analysis is needed to check whether these strategies can be used as the potential platform for developing an efficient vaccine against NE.

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.

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.

Cutaneous xanthogranuloma associated with Klebsiella pneumoniea in a budgerigar (Melopsittacus undulatus).

Budgerigar is a common name for a colorful Australian native bird belonging to the Melopsittacus undulatus species. It is a very familiar pet around the world and its breeding has been grown in Iran. This study was conducted on a 2-year-old budgerigar with a nodular mass on the left wing. Physical examination revealed a firm, round and well-circumscribed mass approximately 1.70 cm in diameter. Radiographs showed a soft tissue mass with no involvement of bony structures. Fine needle aspiration was performed and the sample was cultured. In cultural examination, Klebsiella spp. were isolated in pure culture. Genus and species of the bacteria were confirmed using multiplex polymerase chain reaction. The mass was surgically excised and it was mainly composed of numerous, large lipid-laden macrophages containing abundant vacuolated cytoplasm, extracellular acicular cholesterol clefts and large number of multinucleated giant cells (especially multinucleated Touton giant cells) in the dermis. Finally, a diagnosis of cutaneous xanthogranuloma was made based on histopathological findings.

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.

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.

Ossifying fibroma in a canary (Serinus canaria).

A 1-year-old male canary (Serinus canaria, Jibber breed), with a protuberance on the left tibia was referred to the clinic. The bird was not able to use the affected foot to stand on its perch. Results of radiographs revealed a radio-opaque hard tissue mass. The canary was euthanatized and submitted for necropsy. On histopathologic examination, the mass was diagnosed as an ossifying fibroma tumor. To our knowledge, ossifying fibroma has not been reported in birds so far, and this is the first known report of this tumor in birds.