Domestic Poultry Species Research Articles

Comparison of DNA metabarcoding and morphological diet analysis of lumpfish (Cyclopterus lumpus) as methods of estimating sea lice cleaning efficacy inside commercial Atlantic salmon (Salmo salar) sea cages

Lumpfish (Cyclopterus lumpus) are often used as cleaner fish inside salmonid sea cages for biological control of ectoparasitic sea lice. We compared traditional morphological diet assessments with DNA metabarcoding as methods of estimating lumpfish cleaning efficacy as a function of its size, and of its ingestion of alternative prey. Traditional morphological analysis was performed on 122 ethanol-preserved stomachs. The stomachs were removed in the field - following eDNA protocols – from lumpfish collected from industrial Atlantic salmon sea cages in southern Newfoundland. Morphological stomach analysis gave semi-quantitative estimates of prey counts and/or biomass at the order or family level; it showed that shorter lumpfish ate more gammarid amphipods whereas longer lumpfish ate more formulated feed pellets designed for salmon. No sea lice were confidentially morphologically identified in the lumpfish stomachs. In contrast, COI metabarcoding was able to confidently identify the presence of the salmon louse (Lepeophtheirus salmonis) in 61 of 82 successfully barcoded lumpfish stomachs. Metabarcoding showed that diurnally-migrating krill and gammarid amphipods were present in more stomachs in 2018 whereas formulated feed pellets – identified from COI sequences classified as domestic poultry and non-indigenous fish species - were present in more stomachs in 2020. We argue that, when weighed by the total number of reads, the counts per stomach of the number of Global Estimated Sequence Variants (ESV) classified as a particular prey species could be used to estimate the single-species indices of abundance needed for lumpfish breeding values. The number of ESVs classified as the salmon louse could be used to estimate an index of lumpfish cleaning efficacy. As proof-of-concept, we show that 83% of the variance in the number of Norwegian krill heads observed per stomach was explained by two weighed covariates: 1) Norwegian krill ESV counts and 2) lumpfish stomach contents weight.

An insight into vitamin E and lipid nutrition of the plains-wanderer Pedionomus torquatus.

Vitamin E, as α-tocopherol, is an essential antioxidant protecting the body from free radicals. The vitamin E requirement of managed wildlife species is known to be greater than their wild counterparts, predominantly due to higher dietary lipid content and potentially stressful environments. The plains-wanderer (Pedionomus torquatus, Family Pedionomidae [monotypical]) is a critically endangered, superficially quail-like bird that is the focus of an ongoing captive breeding programme in Australia. It is estimated that plains-wanderers have a high vitamin E requirement (compared with domestic poultry species) to offset a high lipid diet and their naturally flighty temperament. This study therefore aims to gain a greater understanding of the nutritional status and vitamin E requirements of plains-wanderers in managed environments. Total lipid and α-tocopherol intake were quantified for 26 zoo-managed plains-wanderers over a series of diet intake trials in addition to measurement of plasma α-tocopherol and cholesterol concentrations. Plains-wanderers that consumed higher portions of dietary fat had significantly lower circulating α-tocopherol concentrations than birds that consumed lower total dietary fat (p < .001). Additionally, plasma cholesterol concentrations of managed plains-wanderers were found to be significantly greater than all other bird species reviewed, irrespective of Family or feeding type. We also present the first published data quantifying the nutritional makeup of stomach contents of a wild plains-wanderer for use as a potential guide for diet formulation. This study forms a vital foundational insight into the nutritional management of plains-wanderers, but further research is required to understand their dietary habits and cholesterol metabolism.

Rapid detection of avian influenza virus based on CRISPR-Cas12a

BackgroundAvian influenza (AI) is a disease caused by the avian influenza virus (AIV). These viruses spread naturally among wild aquatic birds worldwide and infect domestic poultry, other birds, and other animal species. Currently, real-time reverse transcription polymerase chain reaction (rRT-PCR) is mainly used to detect the presence of pathogens and has good sensitivity and specificity. However, the diagnosis requires sophisticated instruments under laboratory conditions, which significantly limits point-of-care testing (POCT). Rapid, reliable, non-lab-equipment-reliant, sensitive, and specific diagnostic tests are urgently needed for rapid clinical detection and diagnosis. Our study aimed to develop a reverse transcription recombinase polymerase amplification (RT-RPA)/CRISPR method which improves on these limitations.MethodsThe Cas12a protein was purified by affinity chromatography with Ni-agarose resin and observed using sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE). Specific CRISPR RNA (crRNA) and primers targeting the M and NP genes of the AIV were designed and screened. By combining RT-RPA with the Cas12a/crRNA trans-cleavage system, a detection system that uses fluorescence readouts under blue light or lateral flow strips was established. Sensitivity assays were performed using a tenfold dilution series of plasmids and RNA of the M and NP genes as templates. The specificity of this method was determined using H1–H16 subtype AIVs and other avian pathogens, such as newcastle disease virus (NDV), infectious bursal disease virus (IBDV), and infectious bronchitis virus (IBV).ResultsThe results showed that the method was able to detect AIV and that the detection limit can reach 6.7 copies/μL and 12 copies/μL for the M and NP gene, respectively. In addition, this assay showed no cross-reactivity with other avian-derived RNA viruses such as NDV, IBDV, and IBV. Moreover, the detection system presented 97.5% consistency and agreement with rRT-PCR and virus isolation for detecting samples from poultry. This portable and accurate method has great potential for AIV detection in the field.ConclusionAn RT-RPA/CRISPR method was developed for rapid, sensitive detection of AIV. The new system presents a good potential as an accurate, user-friendly, and inexpensive platform for point-of-care testing applications.

BotCl, the First Chlorotoxin-like Peptide Inhibiting Newcastle Disease Virus: The Emergence of a New Scorpion Venom AMPs Family.

Newcastle disease virus (NDV) is one of the most serious contagions affecting domestic poultry and other avian species. It causes high morbidity and mortality, resulting in huge economic losses to the poultry industry worldwide. Despite vaccination, NDV outbreaks increase the need for alternative prevention and control means. In this study, we have screened fractions of Buthus occitanus tunetanus (Bot) scorpion venom and isolated the first scorpion peptide inhibiting the NDV multiplication. It showed a dose dependent effect on NDV growth in vitro, with an IC50 of 0.69 µM, and a low cytotoxicity on cultured Vero cells (CC50 > 55 µM). Furthermore, tests carried out in specific pathogen-free embryonated chicken eggs demonstrated that the isolated peptide has a protective effect on chicken embryos against NDV, and reduced by 73% the virus titer in allantoic fluid. The N-terminal sequence, as well as the number of cysteine residues of the isolated peptide, showed that it belongs to the scorpion venom Chlorotoxin-like peptides family, which led us to designate it "BotCl". Interestingly, at 10 µg/mL, BotCl showed an inhibiting effect three times higher than its analogue AaCtx, from Androctonus australis (Aa) scorpion venom, on NDV development. Altogether, our results highlight the chlorotoxin-like peptides as a new scorpion venom AMPs family.

Human death due to H5N1 amid the COVID-19 pandemic and Mpox outbreak: A call for action.

Human death due to H5N1 amid the COVID-19 pandemic and Mpox outbreak: A call for action.

Protective Efficacy of H9N2 Avian Influenza Vaccines Inactivated by Ionizing Radiation Methods Administered by the Parenteral or Mucosal Routes.

H9N2 viruses have become, over the last 20 years, one of the most diffused poultry pathogens and have reached a level of endemicity in several countries. Attempts to control the spread and reduce the circulation of H9N2 have relied mainly on vaccination in endemic countries. However, the high level of adaptation to poultry, testified by low minimum infectious doses, replication to high titers, and high transmissibility, has severely hampered the results of vaccination campaigns. Commercially available vaccines have demonstrated high efficacy in protecting against clinical disease, but variable results have also been observed in reducing the level of replication and viral shedding in domestic poultry species. Antigenic drift and increased chances of zoonotic infections are the results of incomplete protection offered by the currently available vaccines, of which the vast majority are based on formalin-inactivated whole virus antigens. In our work, we evaluated experimental vaccines based on an H9N2 virus, inactivated by irradiation treatment, in reducing viral shedding upon different challenge doses and compared their efficacy with formalin-inactivated vaccines. Moreover, we evaluated mucosal delivery of inactivated antigens as an alternative route to subcutaneous and intramuscular vaccination. The results showed complete protection and prevention of replication in subcutaneously vaccinated Specific Pathogen Free White Leghorn chickens at low-to-intermediate challenge doses but a limited reduction of shedding at a high challenge dose. Mucosally vaccinated chickens showed a more variable response to experimental infection at all tested challenge doses and the main effect of vaccination attained the reduction of infected birds in the early phase of infection. Concerning mucosal vaccination, the irradiated vaccine was the only one affording complete protection from infection at the lowest challenge dose. Vaccine formulations based on H9N2 inactivated by irradiation demonstrated a potential for better performances than vaccines based on the formalin-inactivated antigen in terms of reduction of shedding and prevention of infection.

Application of Ultraviolet Light for Poultry Production: A Review of Impacts on Behavior, Physiology, and Production

The application of ultraviolet (UV) light in poultry production is garnering increased interest with the drive toward improved poultry welfare and optimized production. Poultry can see in the UV spectrum (UVA wavelengths: 320–400 nm) thus inclusion of these shorter wavelengths may be viewed as more natural but are typically excluded in conventional artificial lights. Furthermore, UVB wavelengths (280–315) have physiological impact through stimulation of vitamin D pathways that can then improve skeletal health. However, better understanding of the effects of UV supplementation must occur before implementation practically. This non-systematic literature review aimed to summarize the impacts of UV supplementation on the behavior, welfare, and production of laying hens, meat chickens (breeders and growers), and other domestic poultry species including directions for future research. The literature demonstrated that UVA light has positive impacts on reducing fear and stress responses but in some research, it significantly increases feather pecking over age during the production phase. UVB light will significantly improve skeletal health, but an optimum duration of exposure is necessary to get this benefit. Supplementation with UVB light may have more distinct impacts on egg production and eggshell quality when hens are experiencing a dietary vitamin D3deficiency, or if they are at the terminal end of production. The relative benefits of UVB supplementation across different ages needs to be further verified along with commercial trials to confirm beneficial or detrimental impacts of adding UVA wavelengths. Further research is warranted to determine whether adding natural light wavelengths to indoor poultry production is indeed a positive step toward optimizing commercial housing systems.

Age-Associated Changes in Recombinant H5 Highly Pathogenic and Low Pathogenic Avian Influenza Hemagglutinin Tissue Binding in Domestic Poultry Species.

Simple SummaryWe evaluated differences in avian influenza H5 Hemagglutinin (HA) tissue binding across age groups using recombinant H5 HA (rHA) proteins. Gene sequences from low pathogenic (LPAIV) (A/mallard/MN/410/2000(H5N2) and an a high pathogenic (HPAIV) (A/Northern pintail/Washington/40964/2014(H5N2)) influenza A virus (IAV) were used to generate the rHA proteins. Respiratory and intestinal tracts from chickens, ducks (Mallard, Pekin, Muscovy), and turkeys of different age groups were used to detect rHA binding with protein histochemistry, which was quantified as the median area of binding (MAB) for statistical analysis. Turkeys had significant differences in the HPAIV rHA binding in the respiratory tract, with younger turkeys having higher levels of binding in the lung compared to the older group. In the intestinal tract, younger turkeys had higher levels of binding compared to the older birds. Using LPAIV, only turkeys had overall significant differences in the respiratory tract MAB, with the older birds having higher levels of binding compared to the younger group. We saw no age-related differences in the overall intestinal tract rHA binding. Age-related differences in rHA binding of the LPAIV and HPAIV rHA demonstrated in this study may partially, but not completely, explain differences in host susceptibility to infection observed during avian influenza outbreaks and in experimental infection studies.The 2014 outbreak of clade 2.3.4.4A highly pathogenic avian influenza (HPAI) led to the culling of millions of commercial chickens and turkeys and death of various wild bird species. In this outbreak, older chickens and turkeys were commonly infected, and succumbed to clinical disease compared to younger aged birds such chicken broilers. Some experimental studies using waterfowl species have shown age-related differences in susceptibility to clinical disease with HPAI viruses. Here, we evaluate differences in H5 Hemagglutinin (HA) tissue binding across age groups, using recombinant H5 HA (rHA) proteins generated using gene sequences from low pathogenic (A/mallard/MN/410/2000(H5N2 (LPAIV)) and a HPAIV (A/Northern pintail/Washington/40964/2014(H5N2)) influenza A virus (IAV). Respiratory and intestinal tracts from chickens, ducks (Mallard, Pekin, Muscovy) and turkeys of different age groups were used to detect rHA binding with protein histochemistry, which was quantified as the median area of binding (MAB) used for statistical analysis. There were species and tissue specific differences in the rHA binding among the age groups; however, turkeys had significant differences in the HPAIV rHA binding in the respiratory tract, with younger turkeys having higher levels of binding in the lung compared to the older group. In addition, in the intestinal tract, younger turkeys had higher levels of binding compared to the older birds. Using LPAIV, similar species and tissues, specific differences were seen among the age groups; however, only turkeys had overall significant differences in the respiratory tract MAB, with the older birds having higher levels of binding compared to the younger group. No age-related differences were seen in the overall intestinal tract rHA binding. Age-related differences in rHA binding of the LPAIV and HPAIV demonstrated in this study may partially, but not completely, explain differences in host susceptibility to infection observed during avian influenza outbreaks and in experimental infection studies.

Probiotic Bacilli Inhibit Salmonella Biofilm Formation Without Killing Planktonic Cells.

Salmonellosis is a foodborne infection caused by Salmonella. Domestic poultry species are one of the main reservoirs of Salmonella, which causes the foodborne infection salmonellosis, and are responsible for many cases of animal-to-human transmission. Keeping backyard chickens is now a growing trend, increasing the frequency of direct contact with the flock and, by consequence, the incidence of Salmonella infections. Bacillus subtilis KATMIRA1933 and Bacillus amyloliquefaciens B-1895 are probiotic bacilli that produce the bacteriocins subtilosin A and subtilin, respectively. The antimicrobial activity of the two strains was determined against the reference strain Micrococcus luteus ATCC 10420. The cell-free supernatant of B. subtilis KATMIRA1933 inhibited biofilm formation by Salmonella enterica subsp. enterica serovar Hadar, Salmonella enterica subsp. enterica serovar Enteritidis phage type 4, and Salmonella enterica subsp. enterica serovar Thompson by 51.1, 48.3, and 56.9%, respectively. The cell-free supernatant of B. amyloliquefaciens B-1895 inhibited the biofilm formation of these Salmonella strains by 30.4, 28.6, and 35.5%, respectively. These findings suggest that the bacillus strains may have the potential to be used as probiotics and antibiotic alternatives for the control of Salmonella in poultry. The number of planktonic cells was unaffected by treatment with the cell-free supernatant. A co-culture of the Salmonella strains with either bacilli showed no signs of growth inhibition, suggesting that it might have been quorum sensing that is affected by the two Bacillus strains.

Susceptibility of turkeys, chickens and chicken embryos to SARS-CoV-2.

The susceptibility of turkeys, chickens and chicken embryos to SARS-CoV-2 was evaluated by experimental infection. Turkeys and chickens were inoculated using a combination of intranasal, oral and ocular routes. Both turkeys and chickens did not develop clinical disease or seroconvert following inoculation. Viral RNA was not detected in oral swabs, cloacal swabs or in tissues using quantitative real-time RT-PCR. In addition, chicken embryos were inoculated by various routes including the yolk sac, intravenous, chorioallantoic membrane and allantoic cavity. In all instances, chicken embryos failed to support replication of the virus. SARS-CoV-2 does not affect turkeys or chickens in the current genetic state and does not pose any potential risk to establish an infection in both species of domestic poultry.

Surveillance and Genetic Characterization of Virulent Newcastle Disease Virus Subgenotype V.3 in Indigenous Chickens from Backyard Poultry Farms and Live Bird Markets in Kenya.

Kenyan poultry consists of ~80% free-range indigenous chickens kept in small flocks (~30 birds) on backyard poultry farms (BPFs) and they are traded via live bird markets (LBMs). Newcastle disease virus (NDV) was detected in samples collected from chickens, wild farm birds, and other domestic poultry species during a 2017–2018 survey conducted at 66 BPFs and 21 LBMs in nine Kenyan counties. NDV nucleic acids were detected by rRT-PCR L-test in 39.5% (641/1621) of 1621 analyzed samples, of which 9.67% (62/641) were NDV-positive by both the L-test and a fusion-test designed to identify the virulent virus, with a majority being at LBMs (64.5%; 40/62) compared to BPFs (25.5%; 22/62). Virus isolation and next-generation sequencing (NGS) on a subset of samples resulted in 32 complete NDV genome sequences with 95.8–100% nucleotide identities amongst themselves and 95.7-98.2% identity with other east African isolates from 2010-2016. These isolates were classified as a new sub-genotype, V.3, and shared 86.5–88.9% and 88.5–91.8% nucleotide identities with subgenotypes V.1 and V.2 viruses, respectively. The putative fusion protein cleavage site (113R-Q-K-R↓F 117) in all 32 isolates, and a 1.86 ICPI score of an isolate from a BPF chicken that had clinical signs consistent with Newcastle disease, confirmed the high virulence of the NDVs. Compared to genotypes V and VI viruses, the attachment (HN) protein of 18 of the 32 vNDVs had amino acid substitutions in the antigenic sites. A time-scaled phylogeographic analysis suggests a west-to-east dispersal of the NDVs via the live chicken trade, but the virus origins remain unconfirmed due to scarcity of continuous and systematic surveillance data. This study reveals the widespread prevalence of vNDVs in Kenyan backyard poultry, the central role of LBMs in the dispersal and possibly generation of new virus variants, and the need for robust molecular epidemiological surveillance in poultry and non-poultry avian species.

Isolation of Arcobacter spp. and identification of isolates by multiplex PCR from various domestic poultry and wild avian species

PurposeThe purpose of the present study was to determine the extent and seasonal prevalence of Arcobacter spp. in domestic poultry and wild birds in the Kars region of Turkey using multiplex polymerase chain reaction (m-PCR).MethodsIn this study, 1570 samples were collected from domestic poultry and wild avian species. The numbers of collected samples were as follows: 182 fecal samples from chickens, geese, and turkeys from family farms in the Kars region in Turkey; 1089 cloacal swab samples from chickens, geese, ducks, turkeys, and quails from family farms in this region; and 299 fecal samples from wild pigeons, crows, and owls in the same region.ResultsArcobacter spp. were isolated from 17.43%, 35.77%, 3.63%, 6.87%, and 3.33% of the cloacal swab samples obtained from geese, ducks, chickens, turkeys, and quails, respectively. In the stool samples, Arcobacter spp. were isolated from 9.62%, 13.33%, and 4% of chicken, goose, and turkey samples, respectively. In wild birds, the isolation rates of Arcobacter spp. were 6.6%, 12.15%, and 0% in pigeons, crows, and owls, respectively. Using m-PCR, among 171 Arcobacter spp. isolates obtained from poultry and wild birds, 67, 78, 24, and 2 were identified as Arcobacter cryaerophilus, Arcobacter butzleri, Arcobacter skirrowii, and Arcobacter cibarius, respectively.ConclusionsBoth poultry and wild avian species exhibited variable rates of Arcobacter species positivity. The presence of Arcobacter spp. in the digestive tracts of healthy poultry and wild birds may serve as a potential reservoir for the dissemination of these microbes in the environment and their transmission to other animals and humans.

Effects of Ultraviolet Light Supplementation on Pekin Duck Production, Behavior, and Welfare.

Simple SummaryDucks, like other poultry species, have the ability to see in the ultraviolet (UV) portion of the light spectrum; however, very little research has explored how UV lighting affects Pekin duck welfare. Therefore, the objective of this study was to determine how providing supplementary UV light in a UV-deficient environment could affect production parameters and behavior. Pekin ducks were reared under either light-emitting diode (LED) bulbs with supplemental UV light (UV) or just LED bulbs (control). The results from this study indicate that Pekin ducks reared under UV light have both lower acute and chronic stress susceptibility and lower fear responses. It was also found that eye morphology can be manipulated with UV light exposure. There were no treatment differences in production parameters such as growth, feed efficiency, tibial mineral composition, visual gait score, or stride length. The results of this study demonstrate the necessity of UV light exposure in duck growout settings and emphasize the need for correct artificial lighting in the poultry industry.Ducks, like other domestic poultry species, can visualize the ultraviolet (UV) portion of the light spectrum; however, the importance of UV light radiation in artificially lit duck growout facilities remains unknown. The objective of this study was to determine the effects of UV light supplementation on Pekin duck production parameters, eye development, stress, and fear. Pekin ducks were reared with light-emitting diode (LED) lights supplemented with UV light or just LED lights (control). There were no differences in body weight (p = 0.32), feed conversion ratio (p = 0.38), or gait score (p = 0.89). Differences in eye morphology were observed, with ducks reared under UV light having narrower (12.3 ± 0.06632 mm; p = 0.010) and lighter (1.46 ± 0.01826 g; p = 0.025) eyes than the control (12.5 ± 0.05583 mm; 1.53 ± 0.02386 g). Ducks reared in UV environments had lower acute and chronic stress susceptibility with lower plasma corticosterone (6317 ± 593.79 pg/mL; p = 0.024), heterophil to lymphocyte ratios (0.43 ± 0.02889; p = 0.035), and composite asymmetry (0.58 ± 0.0298; p = 0.002) than control ducks (9242 ± 1120.7 pg/mL; 0.54 ± 0.04212; 0.76 ± 0.03726 mm, respectively). Ultraviolet ducks had a faster latency for the first head movement during tonic immobility (61.28 ± 9.4863 s, p = 0.026) and required more attempts to induce tonic immobility (1.71 ± 0.07333, p = 0.018) than control ducks (100.7 ± 14.846 s and 1.48 ± 0.06478, respectively). There were no differences in inversion testing (p = 0.91). These results indicate that UV lighting can lower stress and fear responses in Pekin ducks and can therefore increase welfare. Additionally, this study emphasizes the importance of choosing correct artificial lighting for all poultry species.

The quail genome: insights into social behaviour, seasonal biology and infectious disease response

BackgroundThe Japanese quail (Coturnix japonica) is a popular domestic poultry species and an increasingly significant model species in avian developmental, behavioural and disease research.ResultsWe have produced a high-quality quail genome sequence, spanning 0.93 Gb assigned to 33 chromosomes. In terms of contiguity, assembly statistics, gene content and chromosomal organisation, the quail genome shows high similarity to the chicken genome. We demonstrate the utility of this genome through three diverse applications. First, we identify selection signatures and candidate genes associated with social behaviour in the quail genome, an important agricultural and domestication trait. Second, we investigate the effects and interaction of photoperiod and temperature on the transcriptome of the quail medial basal hypothalamus, revealing key mechanisms of photoperiodism. Finally, we investigate the response of quail to H5N1 influenza infection. In quail lung, many critical immune genes and pathways were downregulated after H5N1 infection, and this may be key to the susceptibility of quail to H5N1.ConclusionsWe have produced a high-quality genome of the quail which will facilitate further studies into diverse research questions using the quail as a model avian species.

Analysis of two avian influenza epidemic models involving fractal-fractional derivatives with power and Mittag-Leffler memories.

Since certain species of domestic poultry and poultry are the main food source in many countries, the outbreak of avian influenza, such as H7N9, is a serious threat to the health and economy of those countries. This can be considered as the main reason for considering the preventive ways of avian influenza. In recent years, the disease has received worldwide attention, and a large variety of different mathematical models have been designed to investigate the dynamics of the avian influenza epidemic problem. In this paper, two fractional models with logistic growth and with incubation periods were considered using the Liouville-Caputo and the new definition of a nonlocal fractional derivative with the Mittag-Leffler kernel. Local stability of the equilibria of both models has been presented. For the Liouville-Caputo case, we have some special solutions using an iterative scheme via Laplace transform. Moreover, based on the trapezoidal product-integration rule, a novel iterative method is utilized to obtain approximate solutions for these models. In the Atangana-Baleanu-Caputo sense, we studied the uniqueness and existence of the solutions, and their corresponding numerical solutions were obtained using a novel numerical method. The method is based on the trapezoidal product-integration rule. Also, we consider fractal-fractional operators to capture self-similarities for both models. These novel operators predict chaotic behaviors involving the fractal derivative in convolution with power-law and the Mittag-Leffler function. These models were solved numerically via the Adams-Bashforth-Moulton and Adams-Moulton scheme, respectively. We have performed many numerical simulations to illustrate the analytical achievements. Numerical simulations show very high agreement between the acquired and the expected results.

Aerosol Transmission of Gull-Origin Iceland Subtype H10N7 Influenza A Virus in Ferrets.

Subtype H10 influenza A viruses (IAVs) have been recovered from domestic poultry and various aquatic bird species, and sporadic transmission of these IAVs from avian species to mammals (i.e., human, seal, and mink) are well documented. In 2015, we isolated four H10N7 viruses from gulls in Iceland. Genomic analyses showed four gene segments in the viruses were genetically associated with H10 IAVs that caused influenza outbreaks and deaths among European seals in 2014. Antigenic characterization suggested minimal antigenic variation among these H10N7 isolates and other archived H10 viruses recovered from human, seal, mink, and various avian species in Asia, Europe, and North America. Glycan binding preference analyses suggested that, similar to other avian-origin H10 IAVs, these gull-origin H10N7 IAVs bound to both avian-like alpha 2,3-linked sialic acids and human-like alpha 2,6-linked sialic acids. However, when the gull-origin viruses were compared with another Eurasian avian-origin H10N8 IAV, which caused human infections, the gull-origin virus showed significantly higher binding affinity to human-like glycan receptors. Results from a ferret experiment demonstrated that a gull-origin H10N7 IAV replicated well in turbinate, trachea, and lung, but replication was most efficient in turbinate and trachea. This gull-origin H10N7 virus can be transmitted between ferrets through the direct contact and aerosol routes, without prior adaptation. Gulls share their habitat with other birds and mammals and have frequent contact with humans; therefore, gull-origin H10N7 IAVs could pose a risk to public health. Surveillance and monitoring of these IAVs at the wild bird-human interface should be continued.IMPORTANCE Subtype H10 avian influenza A viruses (IAVs) have caused sporadic human infections and enzootic outbreaks among seals. In the fall of 2015, H10N7 viruses were recovered from gulls in Iceland, and genomic analyses showed that the viruses were genetically related with IAVs that caused outbreaks among seals in Europe a year earlier. These gull-origin viruses showed high binding affinity to human-like glycan receptors. Transmission studies in ferrets demonstrated that the gull-origin IAV could infect ferrets, and that the virus could be transmitted between ferrets through direct contact and aerosol droplets. This study demonstrated that avian H10 IAV can infect mammals and be transmitted among them without adaptation. Thus, avian H10 IAV is a candidate for influenza pandemic preparedness and should be monitored in wildlife and at the animal-human interface.

Characterization of avian influenza H9N2 viruses isolated from ostriches (Struthio camelus)

H9N2 subtype avian influenza viruses (AIVs) have been isolated from various species of wild birds and domestic poultry in the world, and occasionally transmitted to humans. Although H9N2 AIVs are seldom isolated from ostriches, seven such strains were isolated from sick ostriches in China between 2013 and 2014. Sequence analysis showed several amino acid changes relating to viral adaptation in mammals were identified. The phylogenetic analyses indicated that these isolates were quadruple reassortant viruses, which are different from the early ostrich isolates from South Africa or Israel. Most of the ostrich virus carried a human-type receptor-binding property. The chicken experiments showed the ostrich strains displayed low pathogenicity, while they could cause mild to severe symptoms in chicken. Theses strains could efficiently transmit among chickens, and one strain showed higher transmissibility. The virus could not kill mice, and merely replicated in the lung of mice. The ostrich strains could not efficiently transmit between guinea pigs in the direct contact model. These results suggested we should pay attention to the interface between ostrich and other domestic fowl, and keep an eye on this population when monitoring of influenza virus.

Assessment of listing and categorisation of animal diseases within the framework of the Animal Health Law (Regulation (EU) No2016/429): Salmonella infection in poultry with serotypes of animal health relevance (S.Pullorum, S.Gallinarum and S.arizonae).

Salmonella infection in poultry (Salmonella Pullorum, Salmonella Gallinarum and Salmonella arizonae) has been assessed according to the criteria of the Animal Health Law (AHL), in particular criteria of Article 7 on disease profile and impacts, Article 5 on the eligibility of Salmonella to be listed, Article 9 for the categorisation of Salmonella according to disease prevention and control rules as in Annex IV and Article 8 on the list of animal species related to Salmonella. The assessment has been performed following a methodology composed of information collection and compilation, expert judgement on each criterion at individual and, if no consensus was reached before, also at collective level. The output is composed of the categorical answer, and for the questions where no consensus was reached, the different supporting views are reported. Details on the methodology used for this assessment are explained in a separate opinion. According to the assessment performed, Salmonella can be considered eligible to be listed for Union intervention as laid down in Article 5(3) of the AHL. The disease would comply with the criteria as in Sections 4 and 5 of Annex IV of the AHL, for the application of the disease prevention and control rules referred to in points (d) and (e) of Article 9(1). The assessment here performed on compliance with the criteria as in Section 1 of Annex IV referred to in point (a) of Article 9(1) is inconclusive. The main animal species to be listed for Salmonella according to Article 8(3) criteria are all species of domestic poultry and wild species of mainly Anseriformes and Galliformes, as indicated in the present opinion.

Assessment of listing and categorisation of animal diseases within the framework of the Animal Health Law (Regulation (EU) No2016/429): low pathogenic avian influenza.

Low pathogenic avian influenza (LPAI) has been assessed according to the criteria of the Animal Health Law (AHL), in particular criteria of Article 7 on disease profile and impacts, Article 5 on the eligibility of LPAI to be listed, Article 9 for the categorisation of LPAI according to disease prevention and control rules as in Annex IV and Article 8 on the list of animal species related to LPAI. The assessment has been performed following a methodology composed of information collection and compilation, expert judgement on each criterion at individual and, if no consensus was reached before, also at collective levels. The output is composed of the categorical answer, and for the questions where no consensus was reached, the different supporting views are reported. Details on the methodology used for this assessment are explained in a separate opinion. According to the assessment performed, LPAI can be considered eligible to be listed for Union intervention as laid down in Article 5(3) of the AHL. The disease would comply with the criteria as in Sections 3 and 5 of Annex IV of the AHL, for the application of the disease prevention and control rules referred to in points (c) and (e) of Article 9(1). The animal species to be listed for LPAI according to Article 8(3) criteria are all species of domestic poultry and wild species of mainly Anseriformes and Charadriiformes, as indicated in the present opinion.

Molecular Characterization of Some Bacteria Isolated from Broiler Chickens Showing Respiratory Manifestations

Bacterial pathogens play an important role in causing respiratory disease in domestic poultry species, so the present study was carried out for isolation of bacteria from broiler chickens suffering from respiratory manifestations with special reference to molecular characterization of E. coli, Detection of virulence genes of E. coli associated with respiratory system infection was carried out in addition to antibiogram activities and plasmid profile of Pseudomonas aeruginosa from nasal swabs, tracheal swabs and lung samples which were collected from twenty seven broiler flocks at private farm in El-Bohira governorate. A total of 41 samples were subjected to bacterial isolation and identification by using cultural, biochemical, PCR, antibiogram and plasmid profile. Out of 41 samples, 63 isolates were recovered which categorized as 21 (33.3%) E. coli, 17 (26.9%) Pseudomonas aeruginosa, 4 (6.3%) Salmonella Enteritidis, 1 (1.6%) Pasteurella multocida, 10 (15.9%) Corynebacterium species, 3 (4.8%) Staphylococcus aureus and 7 (11.1%) coagulase negative Staphylococcus isolates. The results of amplification of phoA gene (specific for genus of E. coli ) revealed that out of 21 tested isolates, 10 (47.6%) isolates were positive. The results of amplification of virulence genes (tsH, fimH and papC) of E. coli revealed that out of ten isolates of E. coli; of them 3 (30%) isolates were positive for tsH gene, 7 (70 %) isolates were positive for fimH gene and 1 (10 %) isolate was positive for papC gene. The antibiotic resistance pattern showed that Pseudomonas aeruginosa showed high resistance (94.6%) to levofloxacin, streptomycin, ciprofloxacin, ofloxacin, erythromycin, chloramphenicol and neomycin , 76.5% were resistant to norfloxacin but 5.9% of pseudomonas aeruginosa isolates were intermediate susceptible to erythromycin, chloramphenicol and neomycin, (17.7 %) to norfloxacin and (5.9%) were susceptible to levofloxacin, streptomycin, ciprofloxacin, ofloxacin and norfloxacin. Out of 17 Pseudomonas aeruginosa isolates, 15(88, 2%) isolates showed plasmid DNA profile (specific band at 25kbp).