Infectious Laryngotracheitis Research Articles

Health surveillance of a potential bridge host: Pathogen exposure risks posed to avian populations augmented with captive-bred pheasants.

Augmentation of wild populations with captive‐bred individuals presents an inherent risk of co‐introducing novel pathogens to naïve species, but it can be an important tool for supplementing small or declining populations. Game species used for human enterprise and recreation such as the ring‐necked pheasant (Phasianus colchicus) are commonly raised in captivity and released onto public and private wildlands as a method of augmenting naturalized pheasant populations. This study presents findings on pathogen exposure from three sources of serological data collected in California during 2014–2017 including (a) 71 pen‐reared pheasants sampled across seven game bird breeding farms, (b) six previously released pen‐reared pheasants captured at two study sites where wild pheasants occurred and (c) 79 wild pheasants captured across six study sites. In both pen‐reared and wild pheasants, antibodies were detected against haemorrhagic enteritis virus (HEV), infectious laryngotracheitis (ILT), infectious bursal disease virus (IBDV), paramyxovirus type 1 (PMV‐1) and Pasteurella multocida (PM). Previously released pen‐reared pheasants were seropositive for HEV, ILT, and PM. Generalized linear mixed models accounting for intraclass correlation within groups indicated that pen‐reared pheasants were more than twice as likely to test positive for HEV antibodies. Necropsy and ancillary diagnostics were performed in addition to serological testing on 40 pen‐reared pheasants sampled from five of the seven farms. Pheasants from three of these farms tested positive by PCR for Siadenovirus, the causative agent of both haemorrhagic enteritis in turkeys and marble spleen disease of pheasants, which are serologically indistinguishable. Following necropsy, owners from the five farms were surveyed regarding husbandry and biosecurity practices. Farms ranged in size from 10,000 to more than 100,000 birds, two farms raised other game bird species on premises, and two farms used some form of vaccination. Biosecurity practices varied by farm, but the largest farm implemented the strictest practices.

Genetic evolution of Marek's disease virus in vaccinated poultry farms.

Background and Aim:The Marek’s disease virus (MDV) is a neoplastic disease causing serious economic losses in poultry production. This study aimed to investigate MDV occurrence in poultry flocks in the Lower Egypt during the 2020 breakout and genetically characterized Meq, gL, and ICP4 genes in field strains of MDV.Materials and Methods:Forty samples were collected from different breeds from eight Egyptian governorates in 2020. All flocks had received a bivalent vaccine (herpesvirus of turkey FC-126 + Rispens CVI988). However, weight loss, emaciation, reduced egg production, paralysis, and rough/raised feather follicles occurred. Samples were collected from feather follicles, liver, spleen, and nerve tissue for diagnosis by polymerase chain reaction. MDV genetic characterization was then performed by sequencing the Meq, gL, and ICP4 genes of five positive samples representing different governorates and breeds.Results:A total of 28 samples were positive for MDV field strains, while two were related to MDV vaccinal strains. All samples tested negative for ALV (A, B, C, D, and J) and REV. Phylogenetic analysis of the Meq gene of sequenced samples revealed that all MDVs were related to the highly virulent European viruses (Gallid herpesvirus 2 ATE and PC12/30) with high amino acid (A.A.) identity 99.2-100%. Alternatively, there was low A.A. identity with the vaccine strains CVI988 and 3004 (up to 82.5%). These results indicate that further investigation of the efficacy of current Egyptian vaccines is required. The Egyptian strains also harbor a specific mutation, allowing clustering into two subgroups (A and B). By mutation analysis of the Meq gene, the Egyptian viruses in our study had R101K, P217A, and E263D mutations present in all Egyptian viruses. Furthermore, R176A and T180A mutations specific to our strains contributed to the high virulence of highly virulent strains. There were no mutations of the gL or ICP4 genes.Conclusion:Further studies should evaluate the protection contributed by current vaccines used in Egypt.

An unusual case of haemorrhagic bursa in a pullet flock associated with a genogroup 2 infectious bursal disease virus

ABSTRACT An unusual case of infectious bursal disease (IBD) was observed in eight-week-old commercial caged pullets. This flock (House 1) exhibited a one-day spike in mortality. On gross necropsy examination, enlarged, diffusely haemorrhagic bursas were observed. This lesion has been frequently described in cases of very virulent infectious bursal disease virus (vvIBDV). A five-week-old caged pullet flock (House 2) in an adjacent building did not display haemorrhagic bursa lesions. Microscopic examination of bursas from the eight-week-old pullets in House 1 showed marked diffuse haemorrhages and extensive lymphoid necrosis. Histopathology of bursas from the five-week-old pullets in House 2 showed severe, diffuse lymphoid depletion without haemorrhages. IBD ELISA results from birds in House 1 at 9 weeks had a GMT of 6395 and birds in House 2 had a GMT of 82 in the same timeframe. Diagnostic testing for avian influenza virus, Mycoplasma gallisepticum, Mycoplasma synoviae, virulent Newcastle disease virus, infectious bronchitis virus, infectious laryngotracheitis virus, chicken anaemia virus and fowl pox virus were negative. The predicted amino acid sequence of the hypervariable region of VP2 indicated the IBDV observed in both flocks (1/chicken/USA/1300OH/19 from House 1 and 1/chicken/USA/1301OH/19 from House 2) was identical and was not a vvIBDV. Their sequences were similar to a genogroup 2 IBDV from Ontario, Canada (EF138967). No mortality was observed when the 1/chicken/USA/1300OH/19 virus was inoculated into specific-pathogen-free (SPF), four-week-old pullets. Gross and microscopic lesions were observed in bursa tissue, but the bursal haemorrhages observed in the original field case were not reproduced in challenged SPF pullets.

Detection and Molecular Characterization of Avian Infectious Laryngotracheitis Virus Isolated from a Breeder Flock

Detection and Molecular Characterization of Avian Infectious Laryngotracheitis Virus Isolated from a Breeder Flock

Detection of Laryngotracheitis Virus in Poultry Flocks with Respiratory Disorders in Slovenia.

Infectious laryngotracheitis (ILT) is an acute, highly contagious infectious disease of the upper respiratory tract in chickens and other poultry species that causes significant economic losses in countries worldwide. Between 2017 and 2019, seven outbreaks of mild to severe respiratory disorders with high suspicion of ILT occurred in commercial and backyard poultry flocks in Slovenia. In all submissions, infection with ILT virus (ILTV) was confirmed by PCR, which is the first report of ILT in Slovenia. Circulating ILT strains were characterized by the sequence and phylogenetic analysis of two fragments of the ICP4 gene. Four strains—three detected in non-vaccinated flocks and one in a flock vaccinated against ILT—were identical or very similar to the chicken embryo–origin live virus vaccines, and the other three were closely related to Russian, Chinese, Australian, and American field strains and to tissue culture origin vaccine strains. As in other diseases, coinfections with other respiratory pathogens in confirmed ILT cases may cause a more severe condition and prolong the course of the disease. In our study, coinfections with Mycoplasma synoviae (7/7 tested flocks), infectious bronchitis virus (5/5 tested flocks), Mycoplasma gallisepticum (4/7 tested flocks), Ornithobacterium rhinotracheale (3/4 tested flocks), and avian pox virus (1/2 tested flocks) were confirmed, indicating the importance of these pathogens in the occurrence of ILT infections.

US3 Serine/Threonine Protein Kinase from MDV-1, MDV-2, and HVT Differentially Regulate Viral Gene Expression and Replication.

Gallid alphaherpesvirus 2 (GaHV-2), commonly known as Marek’s disease virus type 1 (MDV-1), is an oncogenic avian alphaherpesvirus, and along with its close relatives—Gallid alphaherpesvirus 3 (GaHV-3) or MDV-2 and Meleagrid alphaherpesvirus 1 (MeHV-1) or turkey herpesvirus (HVT)—belongs to the Mardivirus genus. We and others previously showed that MDV-1 US3 protein kinase plays an important role in viral replication and pathogenesis, which could be partially compensated by MDV-2 and HVT US3. In this study, we further studied the differential roles of MDV-1, MDV-2 and HVT US3 in regulating viral gene expression and replication. Our results showed that MDV-2 and HVT US3 could differentially compensate MDV-1 US3 regulation of viral gene expression in vitro. MDV-2 and HVT US3 could also partially rescue the replication deficiency of MDV-1 US3 null virus in the spleen and thymus, as determined by immunohistochemistry analysis of MDV-1 pp38 protein. Importantly, using immunohistochemistry and dual immunofluorescence assays, we found that MDV-2 US3, but not HVT US3, fully compensated MDV-1 US3 regulation of MDV-1 replication in bursal B lymphocytes. In conclusion, our study provides the first comparative analysis of US3 from MDV-1, MDV-2 and HVT in regulating viral gene expression in cell culture and MDV-1 replication in lymphocytes.

The requirement of glycoprotein C (gC) for interindividual spread is a conserved function of gC for avian herpesviruses

We have formerly shown that glycoprotein C (gC) of Gallid alphaherpesvirus 2, better known as Marek’s disease (MD) alphaherpesvirus (MDV), is required for interindividual spread in chickens. Since gC is conserved within the Alphaherpesvirinae subfamily, we hypothesized gC was important for interindividual spread of other alphaherpesviruses. To test this hypothesis, we first generated a fluorescent protein tagged clone of Gallid alphaherpesvirus 3 MD vaccine strain 301B/1 to track virus replication in cell culture and chickens using fluorescent microscopy. Following validation of this system, we removed the open reading frame of 301B/1 gC from the genome and determined whether it was required for interindividual spread using experimental and natural infection studies. Interindividual spread of MD vaccine 301B/1 was abrogated by removal of 301B/1 gC. Rescuent virus in which 301B/1 gC was inserted back into the genome efficiently spread among chickens. To further study the conserved function of gC, we replaced 301B/1 gC with MDV gC and this virus also efficiently spread in chickens. These data suggest the essential function of alphaherpesvirus gC proteins is conserved and can be exploited during the generation of future vaccines against MD that affects the poultry industry worldwide.

Metagenomic Analysis of the Respiratory Microbiome of a Broiler Flock from Hatching to Processing

Elucidating the complex microbial interactions in biological environments requires the identification and characterization of not only the bacterial component but also the eukaryotic viruses, bacteriophage, and fungi. In a proof of concept experiment, next generation sequencing approaches, accompanied by the development of novel computational and bioinformatics tools, were utilized to examine the evolution of the microbial ecology of the avian trachea during the growth of a healthy commercial broiler flock. The flock was sampled weekly, beginning at placement and concluding at 49 days, the day before processing. Metagenomic sequencing of DNA and RNA was utilized to examine the bacteria, virus, bacteriophage, and fungal components during flock growth. The utility of using a metagenomic approach to study the avian respiratory virome was confirmed by detecting the dysbiosis in the avian respiratory virome of broiler chickens diagnosed with infection with infectious laryngotracheitis virus. This study provides the first comprehensive analysis of the ecology of the avian respiratory microbiome and demonstrates the feasibility for the use of this approach in future investigations of avian respiratory diseases.

Pathogenic and Transmission Potential of Wildtype and Chicken Embryo Origin (CEO) Vaccine Revertant Infectious Laryngotracheitis Virus.

Infectious laryngotracheitis (ILT) is an infectious upper respiratory tract disease that impacts the poultry industry worldwide. ILT is caused by an alphaherpesvirus commonly referred to as infectious laryngotracheitis virus (ILTV). Vaccination with live attenuated vaccines is practiced regularly for the control of ILT. However, extensive and improper use of live attenuated vaccines is related to vaccine viruses reverting to virulence. An increase in mortality and pathogenicity has been attributed to these vaccine revertant viruses. Recent studies characterized Canadian ILTV strains originating from ILT outbreaks as related to live attenuated vaccine virus revertants. However, information is scarce on the pathogenicity and transmission potential of these Canadian isolates. Hence, in this study, the pathogenicity and transmission potential of two wildtype ILTVs and a chicken embryo origin (CEO) vaccine revertant ILTV of Canadian origin were evaluated. To this end, 3-week-old specific pathogen-free chickens were experimentally infected with each of the ILTV isolates and compared to uninfected controls. Additionally, naïve chickens were exposed to the experimentally infected chickens to mimic naturally occurring infection. Pathogenicity of each of these ILTV isolates was evaluated by the severity of clinical signs, weight loss, mortality, and lesions observed at the necropsy. The transmission potential was evaluated by quantification of ILTV genome loads in oropharyngeal and cloacal swabs and tissue samples of the experimentally infected and contact-exposed chickens, as well as in the capacity to produce ILT in contact-exposed chickens. We observed that the CEO vaccine revertant ILTV isolate induced severe disease in comparison to the two wildtype ILTV isolates used in this study. According to ILTV genome load data, CEO vaccine revertant ILTV isolate was successfully transmitted to naïve contact-exposed chickens in comparison to the tested wildtype ILTV isolates. Overall, the Canadian origin CEO vaccine revertant ILTV isolate possesses higher virulence, and dissemination potential, when compared to the wildtype ILTV isolates used in this study. These findings have serious implications in ILT control in chickens.

Control, Elimination, and Monitoring of Infectious Laryngotracheitis in a Multiage Commercial Layer Pullet Farm in Canada.

A multiage commercial layer pullet operation with a history of chicken embryo (CEO) modified live infectious laryngotracheitis (ILT) virus vaccination suffered severe ILT breaks in 2017. The initial sequencing revealed that the circulating virus was of vaccine origin. Changes to the timing and dosage of CEO ILT vaccine failed to control the outbreak. The clinical resolution of the outbreak occurred with the transition to a turkey herpesvirus vector vaccine given in hatchery followed by a tissue culture origin (TCO) vaccine given before the onset of symptoms on the farm. The circulating ILT viruses were monitored periodically using next generation sequencing. This site became free of ILT virus within one year after implementing the new vaccination program.

Control, Suppression, and Monitoring of Infectious Laryngotracheitis in a Multiage Commercial Layer Pullet Farm in Canada.

A multiage commercial layer pullet operation with a history of chicken embryo-origin (CEO) modified live infectious laryngotracheitis (ILT) virus vaccination suffered severe ILT outbreaks in 2017. The initial sequencing revealed that the circulating virus was of vaccine origin. Changes to the timing and dosage of CEO ILT vaccine failed to control the outbreak. The clinical resolution of the outbreak occurred with the transition to a turkey herpesvirus vector vaccine given in-hatchery, followed by a tissue culture-origin vaccine given on the farm. The circulating ILT viruses were monitored periodically by next-generation sequencing. This site became free of ILT virus within 1 yr after implementing the new vaccination program.

Research Note: Simultaneous detection of infectious laryngotracheitis virus, fowlpox virus, and reticuloendotheliosis virus in chicken specimens

Infectious laryngotracheitis (ILT), fowlpox (FP), and reticuloendotheliosis are important poultry diseases caused by gallid herpesvirus 1 (ILTV), fowlpox virus (FWPV), and reticuloendotheliosis virus (REV), respectively. Coinfections with ILTV and FWPV occur naturally in chickens, and FP in its more virulent wet form is characterized by diphtheritic lesions and easily confused with ILT. Moreover, the insertion of only partial REV-LTR or a nearly full-length REV into the FWPV genome, located between the ORF 201 and ORF 203, has increased recently in wild-type field FWPV isolates. Therefore, it is critical to detect ILTV, FWPV, REV-integrated FWPV, and REV early and accurately. In this study, we successfully developed a multiplex PCR assay for the simultaneous detection of ILTV, FWPV, REV-integrated FWPV, and REV, and the detection limits was 1 × 54 copies/tube. When used to test clinical samples, the results of the multiplex PCR were in 100% agreement with singleplex PCRs and sequencing. This new multiplex PCR is a simple, rapid, sensitive, specific, and cost-effective method for detection of 4 viruses in clinical specimens.

Marek's disease virus in vaccinated poultry flocks in Turkey: its first isolation with molecular characterization.

Marek's disease (MD) is an important disease of avian species and a potential threat to the poultry industry worldwide. In this study, 16 dead commercial chickens from flocks with suspected MD were necropsied immediately after death. Pathological findings were compatible with MD, and gallid alphaherpesvirus 2 was identified in PCR of spleen samples. Virus isolation was performed in primary cell culture, and partial sequencing of the meq gene of the isolate revealed >99% nucleotide sequence identity to virulent and very virulent plus strains from a number of European countries, placing it in the same subclade of clade III as two virulent Italian strains and a very virulent plus Polish strain as well as virulent strains of geese and ducks. The data reported here indicate that a virulent strain of Marek's disease virus is circulating in Turkey and has not been stopped by the current national vaccination programme.

Gallid Alphaherpesvirus 2 in the Egyptian Turkeys: Molecular Characterization and Establishment of a Universal System for Phylogenetic Classification

Introduction: Gallid alphaherpesvirus 2 (GaHV-2) is a highly contagious oncogenic virus that causes Marek’s disease in chickens and occasionally in turkeys. Among 100 genes identified in GaHV-2 genome, the Meq gene appears to involve viral virulence, oncogenicity, and genetic diversity. Despite the use of Meq gene sequences in phylogenetic classification of GaHV-2 strains circulating in many countries worldwide, no integrated system exists yet. Methods: Turkeys from 2 commercial Egyptian farms were presented with signs of dullness, dehydration, and emaciation. Samples prepared from the internal organs were examined by histopathology and immunohistochemistry. Pools of the internal organs were analyzed by PCR for identification of GaHV-2, avian leucosis virus, and reticuloendotheliosis virus. The Meq gene of an Egyptian strain was sequenced and analyzed in comparison to 40 reference strains for generation of a universal system for phylogenetic classification of GaHV-2 strains. Results: Gross and histopathological examination revealed grayish-white soft masses in the internal organs characterized by diffuse infiltration of pleomorphic neoplastic cells. All lymphoma cells were identified as T-lymphocytes of CD3+ phenotype. Samples of both farms were only positive for GaHV-2 by PCR. Sequence analysis of the Meq gene has classified the current turkey strain as related to the Egyptian strains identified in chicken in 2012. A universal phylogenetic system for classification of GaHV-2 strains into 4 clusters was proposed. The vaccine strains were all grouped in cluster 2, and most of the classical American strains belonged to cluster 4. Cluster 1 was further divided into 3 subclusters (1.1–1.3). Conclusion: GaHV-2 was identified in turkeys for the first time in Africa and the Middle East. Sequence analysis of the Meq gene of the Egyptian strain along with a wide array of the global strains has enabled the construction of a novel phylogenetic classification system.

Pathobiological and molecular studies on infectious Laryngotracheitis virus (ILTV) infection in commercial chicken flocks in India

Infectious laryngotracheitis (ILT) is one of the highly infectious and contagious diseases of chickens caused by Gallid Herpesvirus 1, a member of the genus Iltovirus. It is an economically important disease in major poultry producing countries, including India. Despite India having large population of poultry birds, very limited information is available on the disease magnitude and molecular epidemiology. We screened 560 tissue samples collected from necropsied cases of tracheitis and conjunctivitis to diagnose ILT by histopathology, immunohistochemistry and polymerase chain reaction. Out of 560 samples, ILT was diagnosed in 90 (16.07%) cases by histopathology; however, polymerase chain reaction detected ILT virus (ILTV) in 128 (22.85%). Immu-nohistochemistry of representative PCR positive tissues (n=30) from trachea, lungs and conjunctiva detected viral antigen in the lining or denuded epithelial cells in 76.66% (23/30) cases. Analysis of age wise occurrence of ILT cases in our study revealed 47.65% in adult birds over 18 weeks of age, 35.15% in grower birds aged between 7 and18 weeks of age, 17.18% in chicks aged less than 6 weeks. Nucleotide sequence of gene encoding viral thymidine kinase revealed involvement of vaccine and virulent strains of ILTV in the investigated cases. Based on our findings and recent publications, we conclude that there is resurgence or re-emergence of ILT in the chicken flocks in different parts of India, indicating a possible biosecurity breach. Further genetic characterization of virus by whole genome sequencing technologies would decipher the genotype circulating in India, and help to design suitable native vaccine candidate(s).

Detection and molecular characterization of a highly oncogenic Marek's disease virus from vaccinated hens in Turkey

Marek's disease (MD) is a highly contagious neoplastic disease of chickens associated with economic losses, often due to visceral lymphomas. The etiological agent is MD virus serotype1 (MDV-1), also called Gallid alphaherpesvirus 2 (GaHV-2). Despite intensive vaccination, MDV is constantly evolving and maintaining its presence in the world. The aim of this study was to genetically analyze a highly oncogenic MDV/Tur/2019 strain obtained from a poultry farm in Turkey's Elazig province in 2019. Genes associated with viral pathogenicity and oncogenicity Marek's EcoRI-Q-encoded protein (MEQ), phosphoprotein-38 (pp38), and viral interleukin 8 (vIL-8) were selected for this purpose. The vIL-8 nucleotide sequence showed high similarity (100% identity) to some European (EU-1, Polen 5) and Asian (03 India, GADVASU-M2) MDV strains. The pp38 nucleotide sequence showed high similarity (100% identity) to some American (CU-2, JM/102W, RB1B) and European (MD70/13, ATE2539) MDV strains. There were no disrupted four-proline molecules (PPPP) within the transactivation domain of the MEQ. However, according to phylogenetic results, the MDV/Tur/2019 strain was included in cluster 2a alongside European MDV strains (Polish, Hungarian, Italian) with very virulent and very virulent plus pathotypes. In conclusion, we believe that the MDV/Tur/2019 strain obtained from turkey herpesvirus (HVT)-vaccinated chickens has a very virulent or very virulent plus pathotype. Although this result provides some clues regarding the virulence of this strain, in vivo studies are needed to achieve exact pathotyping. Further, combination of HVT with the CVI988 strain should be used for vaccination to provide the best protection, as highly pathogenic MDV strains can break sterile immunity against the HVT vaccine. Keywords: GaHV-2; Marek's disease; oncogenes; Turkey.

Evaluation of actions of the official veterinary service to mitigate outbreaks of infectious laryngotracheitis and improve biosecurity on laying hen farms

ABSTRACT: Infectious laryngotracheitis (ILT), caused by an Alphaherpesvirus (Gallid herpesvirus-1; GaHV-1), has been noticed in the region of the Terras Altas da Mantiqueira, Minas Gerais. From 2010 to 2018, the “Serviço Veterinário Oficial” (SVO) of the “Instituto Mineiro Agropecuário” (IMA), implemented measures to prevent spread of the virus to other regions and control the disease in the area. Due to the close proximity and consequent epidemiological link among farms, the region was considered a unique epidemiological unit. To check the efficiency of the ILT control measures, we carried out: (1) a seroepidemiological survey, (2) questionnaires for evaluating biosecurity measures; and (3) an evaluation of the influence of farm population density on the occurrence of ILT. In 2016, 2017, and 2018, ILT was investigated using epidemiological and clinicopathological methods, along with GaHV-1 molecular detection. Serological survey was carried out on 24 farms in the quarantined region and on 13 farms from other regions of the state. In 2010 and 2018, questionnaires were applied to collect data and determine indicators of biosecurity practices in all farms of the quarantined area. The differences were then assessed (Wilcoxon’s p<0.05). The results indicated positive serology throughout the region, although only on four farms (16.6%) the chickens have clinical signs, macroscopic and histological lesions of ILT. The prevalence of viral infection increased from 2016 (27%) to 2017 (50%) and was higher in farms with a high stock density (p=0.033). No disease, virus or antibodies were detected in the farms outside of the quarantined area. Although the biosecurity indicators had improved on all farms in the quarantined area (p<0.05), the virus was active and circulating in the region. The contingency measures have contained the outbreak, but biosecurity practices are paramount in the control of new outbreaks. Official control will be maintained in the region, including surveillance of new cases and biosecurity procedures to mitigate the risk of the virus reaching other regions.

Indicators and risk factors of infectious laryngotracheitis in layer hen flocks in Algeria.

Background and Aim:Since 2017, there have been epidemics with respiratory disorders in the laying hen farms in Algeria, as signs and lesions, respiratory difficulties, and hemorrhagic tracheitis, which closely like laryngotracheitis. This study aimed to analyze the epidemiological, serological, and clinical indicators, as well as the risk factors, of infectious laryngotracheitis (ILT) in layer hen flocks in Algeria.Materials and Methods:A total of 1728 layer hens were sampled randomly from 48 poultry houses. Blood samples were collected from each hen at the wing vein area, and an indirect enzyme-linked immunosorbent assay was done using an IDvet® kit.Results:The flocks showed 56.25% seroprevalence. Clinical signs and gross lesions of ILT suspect cases included respiratory signs characterized by hemorrhagic tracheitis and sinusitis; conjunctivitis; egg drop; and a low mortality rate varying from 5% to 20%. Statistical analyses showed the effect of risk factors on the seropositivity for ILT in 48 layer flocks. When the vaccination was not applied, flocks were significantly more seropositive by 54% (odds ratio OR=1.54, p=0.01) compared to vaccinated flocks. Furthermore, flocks with poor hygiene were more seropositive by 68% (OR=1.68, p=0.002) compared to those with good hygiene. Finally, flocks with decreased egg production between 10% and 30% were significantly more seropositive by 42% (OR=1.42, p=0.04) than those with egg production >30%.Conclusion:The serological survey revealed anti-ILT virus antibodies, signifying the circulation of this virus in layer hen farms in Algeria. Correct vaccination protocol, strict biosecurity measures, rapid diagnosis, and detection of latent carriers are necessary to control and eradicate the disease from layer farms.

Циркуляція вірусу інфекційного ларинготрахеїту в птахівничих господарствах України в 2009–2020 роках

Циркуляція вірусу інфекційного ларинготрахеїту в птахівничих господарствах України в 2009–2020 роках

Recombinase-aided amplification–lateral flow dipstick assay—a specific and sensitive method for visual detection of avian infectious laryngotracheitis virus

The purpose of this study was to explore a specific, simple, and sensitive method for diagnosis of avian infectious laryngotracheitis virus. Recombinase-aided amplification (RAA) and lateral flow dipstick (LFD) were combined for labeling the optimized RAA probe with 6-carboxyfluorescein (FAM) and the 5′-end of the downstream primer with biotin, respectively. By optimizing the reaction time, temperature, and primer concentration of RAA, a RAA–LFD assay, which could be used for detection of infectious laryngotracheitis, was established. After the specificity and sensitivity test, the target gene fragments could be amplified by RAA–LFD assay in 20 min under isothermal conditions (37°C), and the amplification products could be visually observed and determined by LFD within 3 min. There was no cross-reaction with nucleic acids of other avian pathogens, the lowest detectable limit of RAA–LFD was 102 copies/μL, and the sensitivity of this method was 100 times higher than that of conventional PCR with the lowest detectable limit of 104 copies/μL. The results showed that RAA–LFD assay was highly sensitive, easy to use, and more suitable for clinical detection.