Anticoccidial effects of tannin-based herbal formulation (Artemisia annua, Quercus infectoria, and Allium sativum) against coccidiosis in broilers.

Background: Avian coccidiosis is considered among the infectious disease of high cost in the poultry industry. Herbal extracts are safe and reliable substitute anticoccidial drugs for chemical feed additives as they do not sequel to drug resistance and tissue remnants. Objective: The current study aimed to assess the anticoccidial effect of an herbal complex of 3 plants (Artemisia annua, Quercus infectoria, and Allium sativum) in broiler chickens compared to toltrazuril anticoccidial. Methods: This experiment used one hundred twenty broiler chickens and divided them into four equally numbered groups. All the groups, except group (D), were experimentally infected with mixed Eimeria spp. (E. tenella, E. maxima, E. necatrix and E. brunetti) on day 14. Group (A) was treated with a herbal mixture, containing 75% Quercus infectoria with a minimum of 30% total tannin, 16% Artemisia annua with a minimum of 0.02% artemisinin, and 9% Allium sativum with a minimum of 0.4% total phenol contents. Group (B) was treated with toltrazuril. Group (C) did not have any treatment. Group (D) was healthy all the experiment period as a negative control group. During a 42-day breeding period, the study examined clinical signs, weight gains, feed conversion ratio, lesions scoring, casualties, and the number of oocysts in different bird groups. Results: Group (D) showed the most significant weight gain, indicating the economic damage caused by coccidiosis. The best feed conversion ratio was observed in the unchallenged group, and coccidiosis negatively affected it in other groups. Clinical signs of dysentery, diarrhea, and lethargy were seen post-challenge but improved with treatment. Group (D) showed no losses; others had casualties and coccidiosis lesions. Lesion scores were lowest in the group (D), and the herbal mixture improved performance. The herbal mixture and toltrazuril reduced oocyst counts in feces earlier than the untreated group. Conclusion: In conclusion, the anticoccidial activity of the mentioned herbal complex recommends its use as an alternative anticoccidial agent to chemotherapeutic drugs for controlling coccidiosis.

Fluctuation of the prevalence of beak and feather disease virus in captive psittacines in Iran.

Beak and feather disease virus (BFDV) is one of the most life-threatening viral agents infecting parrot species. In this study, we investigated fluctuation in the prevalence of beak and feather disease virus (BFDV) in captive psittacines in Iran. Two series of feather samples from different psittacine species, received between July 2019 and July 2021 (n = 1009) and between July 2021 and July 2022 (n = 2020), were examined for the presence of BFDV using the PCR method, and the host species distribution and temporal prevalence of BFDV within populations were calculated. The results showed a total viral prevalence of 26.86% and 26.88% within sample series 1 and 2, respectively. By examining both sample series, the prevalence of BFDV was found to be  the highest (P < 0.05) in Nymphicus hollandicus and the lowest (P < 0.05) in Psittacus erithacus, Myiopsitta monachus, Pyrrhura molinae, and Aratinga solstitialis. The viral prevalence was significantly higher (P < 0.05) within the series 1 than the series 2 samples only in Nymphicus hollandicus. Within series 2, the viral prevalence was significant (P < 0.05) in samples from Nymphicus hollandicus collected in March. This study indicates significant prevalence of BFDV in captive Nymphicus hollandicus populations and suggests that the fluctuation in the prevalence of BFDV could be due to the combined influence of host-species and temporal factors.

Fungal isolation and identification from parrot excreta in northeast Iran: A threat to human health.

BACKGROUND: One of the environmental sources of pathogenic fungi is parrot droppings. OBJECTIVES: Thus, this work aimed to investigate fungal contamination in parrots' droppings. METHODS: Seventy-nine parrot droppings including Cockatiel, Cockatoo, Green check conure, Love bird, Budgerigar, African grey parrot, Alexandrine parakeet, Amazon parrot, Yellow crown parakeet, and Macaw were sampled, and suspended 1:10 in saline solution and then 0.5 mL of supernatant was cultured. The fungi were identified by standard mycological techniques. RESULTS: Fungal contamination was detected in 66 samples (83.54%) out of 79 samples. Yeast and mould fungi were respectively isolated from 44 samples (55.69%) and 36 samples (45.56%) out of 79 samples. Overall 105 fungal isolates were isolated from parrot excreta. Cryptococcus neoformans (17.14%), Rhizopus spp. (10.47%), Rhodotorula spp. and Aspergillus niger (6.66%), and Penicillium spp. (5.71%) were the most fungi isolated from faecal samples. CONCLUSIONS: The evidence from this study suggests that the rate of fungal contamination in parrots' excreta was high. Keeping parrots in the house and their close contact with humans can double the importance of these contaminations and provide the basis for the transmission of contamination to humans. Therefore, the accumulations of parrot faeces for long periods suggest a potential threat to public health.

Evaluation of therapeutic effects of an herbal mixture (Echinacea purpurea and Glycyrrhiza glabra) for treatment of clinical coccidiosis in broilers.

BACKGROUND: Avian coccidiosis is thought to be one of the most expensive infectious diseases in the poultry industry. OBJECTIVES: Safe and alternative anti-coccidial drugs are herbal extracts because they do not result in tissue residue and drug resistance. The objective of the present study was to evaluate the anti-coccidial effect of the herbal mixture, a complex of two plants (Echinacea purpurea, Glycyrrhiza glabra) in broiler chickens in comparison with toltrazuril. METHODS: One hundred twenty broiler chickens were used in this experiment and divided into 4 equally numbered groups. All the groups, except Group D, were experimentally infected with mixed Eimeria spp. (E. Tenella, E. maxima, E. necatrix and E. brunetti) on day 14. Group A treated with an herbal mixture [Glycyrrhiza glabra Extract 5% (standardised to 5.4% glycyrrhizic acid) and Echinacea purpurea Extract 2% (standardised to 4% total phenolic content based on chlorogenic acid); Coxinin-EC® ; Shamim Teb Sepid Giti]. Group B treated with toltrazuril. Group C was experimentally infected with mixed Eimeria spp. but they did not have any treatment, this group was our positive control. Performance indices, faecal oocyst excretion, and intestinal lesion score were determined during the experiment. RESULTS: Positive control group had the poorest results and more mortality than other groups. Group D was not infected and was healthy all the experiment period. Treatment with herbal complex significantly reduced the negative performance and pathogenic effects associated with Eimeria spp. at a level that was comparable with toltrazuril. CONCLUSIONS: In summary, the anti-coccidial activity of the studied herbal complex suggests its use as an alternative anti-coccidial agent to chemotherapeutic drugs for controlling coccidiosis in poultry. HIGHLIGHTS: -Coccidiosis is an important infectious disease that causes serious financial loss to the poultry industry. -Chemical anti-coccidial drugs and vaccines are the main control strategies to combat the disease. However, these tools have some constraints. -Herbal remedies are suitable alternatives to chemical compounds for control of losses associated with coccidiosis in poultry. -An herbal mixture (Echinacea purpurea, Glycyrrhiza glabra) has promising effects for controlling of coccidiosis in broiler chickens.

Ameliorative role of trans-ferulic acid on induced oxidative toxicity of rooster semen by ß-cyfluthrin during low temperature liquid storage.

Current study was designed to evaluate the effects of ß-cyfluthrin, as a toxicant substance, and trans-ferulic acid (trans-FA), as a protective agent, on different parameters of rooster semen upon liquid storage. For this purpose, semen samples of roosters (Ross 308, n = 10, 32-wk-old) were collected twice a week. Good quality samples (≥70% progressive motion) were diluted, pooled and then divided for the purposes of the study. In the first experiment, motility of spermatozoa was evaluated following exposure to different concentrations of ß-cyfluthrin (1, 2.5, 5, 10, 20, 40, and 80 µM) at 0, 24, and 48 h of storage. In the second experiment, constant doses of ß-cyfluthrin (10 µM) alone or in combination with trans-FA (10, 25 mM) were assessed on motility and viability of spermatozoa at 0, 24, and 48 h time points. Moreover, amounts of malondialdehyde (MDA), total antioxidant capacity (TAC), total nitrate-nitrite, total hydroperoxide (HPO), and activity of superoxide dismutase (SOD) were evaluated in the homogenate of spermatozoa-diluent at studied time points. Results of the first experiment showed that amounts of ß-cyfluthrin greater than 5 µM, significantly reduced the motility of spermatozoa at 24 and 48 h of storage (P < 0.05). The second experiment demonstrated that, trans-FA especially at 10, 25 mM doses restored the motility and viability of spermatozoa compared to ß-cyfluthrin treated group (P < 0.05). Amounts of MDA (10, 25 mM), hydroperoxide (10, 25, and 50 mM), and nitrate-nitrite (10, 25, and 50 mM) were lower and TAC (10 and 25 mM) were greater in trans-FA + ß-cyfluthrin treated groups compared to ß-cyfluthrin alone treated samples (P < 0.05). However, activity of SOD did not show significant changes by the treatment (P > 0.05). It seems that trans-FA could ameliorate toxic effect of ß-cyfluthrin via reduction of peroxidative (as evident by measurement of MDA) and nitrosative (as evident by measurement of nitrate-nitrite) reactions over cold preservation of rooster semen.

Antimicrobial resistance and virulence genes in the Escherichia coli isolates obtained from ostrich.

Escherichia coli (E.coli) is a normal intestinal microflora of birds, including ostrich. However, some strains are pathogenic to ostrich. There is a lack of knowledge concerning the differences between commensal and pathogenic E.coli isolates of ostrich. This research aimed to characterize the antimicrobial susceptibility profile and virulence gene content of presumptive commensal and pathogenic E.coli isolates of ostrich. A total of 129 bacterial isolates (commensal strains no. = 45; pathogenic strains no. = 84) were obtained from ostriches. The resistance profile of these isolates was investigated by the disk diffusion method and PCR. Also, the strains were screened for virulence-associated genes of avian pathogenic E.coli (APEC). The study showed that the highest and the lowest antimicrobial resistance were against oxytetracycline and gentamicin, respectively. Oxytetracycline, florfenicol, and streptomycin resistance rate in pathogenic isolates were higher than commensal ones (p < 0.05). Also, tetA, blaTEM, and aac(3)-IV resistance genes were more prevalent in pathogenic than commensal isolates (p < 0.05). More than half of the isolates had no virulence-associated genes. The multiplex PCR results showed that irp2 gene was more prevalent in pathogenic than commensal E.coli (p < 0.05). Nevertheless this was not the case with the other genes. Our results indicated a low frequency of antimicrobial resistance and virulence genes in E.coli isolates of ostriches. Antimicrobial susceptibility profile and virulence gene content of E.coli isolates of ostriches differ between presumptive commensal and pathogenic strains. However, more analyses are needed to discriminate these isolates.

Outbreak of avian mycobacteriosis in a commercial turkey breeder flock.

Avian mycobacteriosis (AM) is a chronic and contagious disease of pet birds, captive exotic, wild and domestic fowl, and mammals. Mycobacterium avium subsp. avium is the most common cause of AM in poultry. For the first time, we report a chronic outbreak of AM in an Iranian breeder flock of 250 45-week-old turkeys (Meleagris gallopavo) with a morbidity and mortality rate of 91.6% and 80%, respectively. A well-defined clinical feature of the outbreak included a progressive weight loss, decreased egg production, listlessness, and lameness. Tuberculous nodules were seen on liver, spleen, ovary, and ribs. Granulomatous inflammation and acid-fast bacilli were confirmed by using Ziehl-Neelsen method on hepatic lesions. M. avium subsp. avium was identified by polymerase chain reaction techniques based on the presence of 16S ribosomal RNA gene and insertion elements IS1245 and IS901. In this report, we not only describe the epidemiological, pathological, and molecular characteristics of the outbreak in detail, but we also discuss multiple factors influencing the introduction and development of AM critically. In this case, wild feral pigeons might have been the source of infection, but further molecular-epidemiology studies are needed to understand the role of wild birds in the persistence and transmission of Mycobacterium.RESEARCH HIGHLIGHTS First report of avian mycobacteriosis in an Iranian commercial turkey flock is described in detail.Risk factors intrinsic to the bird and mycobacteria, as well as extrinsic factors influencing the introduction and development of avian mycobacteriosis in birds, are critically discussed.

Quercetin attenuates H2 O2 -induced toxicity of rooster semen during liquid storage at 4°C.

Current study was carried out to examine the protective effects of quercetin against toxicity induced by hydrogen peroxide in rooster semen in vitro. Semen samples were collected from ten roosters (Ross 308 broiler breeder males, 32 weeks old) twice a week by abdominal massage method. Samples with ≥70% progressive motility were selected, pooled, diluted and used for the study. Experimental groups consisted of negative control, control that received solvent of quercetin, H2 O2 (40 µM) and combination groups which incubated with constant dose of H2 O2 (40 µM) plus various levels of quercetin (20, 40 and 80 µM). Measurement of total hydroperoxide (HPO), malondialdehyde (MDA), nitric oxide (NO), total antioxidant capacity (TAC) and superoxide dismutase activity as well as routine sperm tests were done at 0, 24 and 48 hr of storage at 4°C. Results revealed that exposure to hydrogen peroxide significantly increased HPO (138.43 ± 7.32 vs. 66.08 ± 3.97 µmol/g protein), MDA (7.21 ± 0.08 vs. 5.71 ± 2.16 µmol/g protein) and NO (0.367 ± 0.013 vs. 0.215 ± 0.011 µmol/g protein) levels and decreased sperm progressive motility (27.28 ± 1.21 vs. 47.49 ± 1.29%), and amounts of TAC (11.49 ± 0.39 vs. 15.70 ± 0.79 mmol/g protein) compared to control at 24 hr (p < 0.05). Changes at mentioned variables were repeated at 48 hr of storage. Also, co-administration of quercetin (especially at 40 and 80 µM) with hydrogen peroxide restored the toxic effects of hydrogen peroxide on rooster semen parameters such as primary and secondary lipid peroxidative indicators and other evaluated variables. The study concluded that rooster semen enrichment with quercetin would protect lipid peroxidative and nitrosative hydrogen peroxide-mediated damage during cold liquid storage of rooster semen.

Evaluation of linoleic acid on lipid peroxidative/antioxidative parameters, motility and viability of rooster spermatozoa during cold storage.

Sustainable reduction in semen quality due to detrimental effects of primary and secondary peroxidative products was occurred during liquid storage. The objective of the current experiment was to explore the influence of bovine serum albumin conjugated linoleic acid (LA) on the rooster spermatozoa routine tests and lipid peroxidative/antioxidative levels stored at 4 °C over 48 h. For this purpose, collected ejaculates (≥ 80% progressive motile spermatozoa) pooled and extended with the phosphate buffer medium without (control) or enriched with different amounts of LA (0.125, 0.25 or 0.50 mM). Contents of total antioxidant status (TAS) and thiobarbituric reactive substances (TBARS) were measured separately in the medium and spermatozoa, as well as percent of viability and motility at 0, 24 and 48 h intervals. Viability was not affected by treatment during the study intervals (P > 0.05). While, higher motility was recorded in LA 0.50 mM group (77.98 ± 1.89 and 57.02 ± 2.45) compared to the control group (68.78 ± 1.29 and 45.09 ± 1.86) at 24 and 48 h, respectively (P < 0.03). Amounts of TBARS in medium and spermatozoa were lower in LA 0.25 and 0.50 groups compared to the control at 48 h (P < 0.01). Moreover, TAS levels of medium and spermatozoa were lower in control samples compared to LA treated groups at 48 h (P < 0.03). Because of the ability of the LA to lowering the quantities of lipid peroxidation index and improving motility especially at 0.5 mM levels, it can be proposed as an additive during liquid storage of rooster semen.

Molecular characterization of Mycoplasma synoviae isolated from broiler chickens of West Azarbaijan province by PCR of vlhA gene.

Mycoplasma synoviae (MS) is a pathogen responsible for respiratory and locomotor disorders and causes major economic losses in poultry industry. Early and accurate diagnosis of MS infection plays a major role in control of the infection. This study was conducted to characterize Iranian field isolates of MS isolated from broiler chickens of West Azarbaijan province (Northwest of Iran), and differentiate them from vaccine strain MS-H. Two encoding genes, 16S rRNA and vlhA were employed. PCR results using primers related to 16s rRNA and vlhA genes were analyzed and compared. Out of 21 field samples, eight samples (38.0%) were positive using both sets of primers. Amplified products of vlhA gene were sequenced for MS strain identification. The results showed that Iranian field isolates of MS had high nucleotide and amino acid similarity. Iranian field isolates were distinct from vaccine strain MS-H. Results presented in this study showed that characterization of field isolates of MS by sequencing of vlhA gene and is beneficial for strain typing and differentiating them from vaccine strain. To our knowledge, this is the first study characterizing vlhA gene of MS isolates from broiler chickens in the West Azarbaijan province.

Seroprevalence of avian influenza (H9N2) in broiler chickens in Northwest of Iran.

OBJECTIVE: To demonstrate seroprevalence of avian invluenza (H9N2) subtybe in broiler chickens in Northwest of Iran. MATERIALS: A total of 310 blood samples were collected from 25 broiler flocks in slaughterhouses of West Azarbayjan, Iran. Serum samples were subjected to haemagglutination inhibition test. RESULTS: The test showed 40.6% of positive serums. Mean antibody titer of avian influenza virus differed between geographical locations in this survey. CONCLUSIONS: High prevalence of avian influenza virus antibodies in serum of birds emphasize that avian influenza has an important role in respiratory complexes in broiler chickens in this region, and probably throughout Iran. Biosecurity measures, monitoring and surveillance programs, and to some degree vaccination are effective tools to prevent introduction of H9N2 infection and its economic losses.