Layer Flocks Research Articles

Occurrence of Marek's Disease in Poland on the Basis of Diagnostic Examination in 2015-2018.

IntroductionMarek’s disease (MD) is a tumourous disease caused by Marek’s disease virus (MDV) and most commonly described in poultry. The aim of the study was to determine the occurrence of Marek’s disease virus infections in Poland and analyse clinical cases in the years 2015–2018.Material and MethodsThe birds for diagnostic examination originated from 71 poultry flocks of various types of production. Birds were subjected to anatomopathological examination post mortem, during which liver and spleen sections and other pathologically changed internal organs were taken. These sections were homogenised with generally accepted methods, then total DNA was isolated and amplified with a real-time PCR. A pair of primers complementary to the MDV genome region encoding the meq gene were used.ResultsMDV infection was found predominantly in broiler chicken flocks (69.01%), and also in layer breeder (9.85%) and commercial layer flocks (7.04% each).ConclusionThe results of research conducted in the years 2015–2018 clearly indicate that the problem of MDV infections is still current.

Phenotypic and genomic analyses of agonistic interactions in laying hen lines divergently selected for feather pecking

In poultry, aggressive pecking and threatening are normal agonistic behavior patterns which serve to establish a social hierarchy. As agonistic behavior is a stressor for animals, its excessive occurrence is undesired in layer flocks. Feather pecking is a longstanding serious problem in layer flocks and its relationship to agonistic behavior is still not clear. Therefore, phenotypic and genomic analyses of the agonistic and feather pecking behavior of two laying hen lines divergently selected for high and low feather pecking were conducted. The hens were phenotyped for the active traits aggressive pecks delivered (APD)11Aggressive pecks delivered (APD), threats delivered (TD)22Threats delivered (TD), feather pecks delivered (FPD)33Feather pecks delivered (FPD) and the passive traits aggressive pecks received (APR)44Aggressive pecks received (APR), threats received (TR)55Threats received (TR) and feather pecks received (FPR)66Feather pecks received (FPR). Indices were built by subtracting the passive traits from the respective active traits to obtain the aggression index, the threat index, and the feather pecking index. As all three behavior patterns in their excessive manifestations are undesired, the index-traits Activity and Passivity were also defined by combining each the active and passive traits. The results showed that FPD is significantly positive correlated with APD and TD in both lines, but with higher coefficients in the high feather pecking line. The average amount of FPR in both lines is nearly the same and no correlation was found between FPR and FPD, APD or TD in any of the lines. The active traits and the feather pecking index showed medium heritabilities, whereas the heritability was negligibly small for the other traits. GWAS revealed four nominal significant (p ≤ 5*10−5) SNPs for APD on chromosome 6, the same four and three additional SNPs on chromosome 8 for Activity and three SNPs on chromosome 1 for the feather pecking index. It is concluded, that selection on high feather pecking leads to an increase of agonistic behavior. The correlation probably depends on the phase of establishing the social hierarchy in which the hens in a newly formed group are at the time of observation, and might disappear, after a stable ranking is established. The reception of feather pecking is similar in both lines. GWAS revealed that TD, APD, Activity and the feather pecking index seem to be typical quantitative traits with associated regions for the latter three which have slightly greater effects on these traits than other regions in the genome.

Identification, HPG2 Sequence Analysis, and Antimicrobial Susceptibility of Avibacterium paragallinarum Isolates Obtained from Outbreaks of Infectious Coryza in Commercial Layers in Sonora State, Mexico.

This is the first extensive report on the identification and characterization of Avibacterium paragallinarum (AVP) isolates obtained from outbreaks of infectious coryza (IC) in IC-vaccinated layer flocks from Sonora State in Mexico. Isolates obtained from IC outbreaks during the years 2007, 2014, 2015, 2017, and 2019 were identified by conventional PCR test and 16S rRNA gene analysis, serotyped by Page serotyping and genotyped by the recently described partial sequence analysis of the HPG2 region. Furthermore, antimicrobial susceptibility profiles were determined by a recently improved minimal inhibitory concentration (MIC) test. The conventional PCR test and the 16S rRNA analyses confirmed the isolates as AVP. Serotyping results showed the involvement of isolates belonging to serotypes A, B, and C in the IC outbreaks. Genotyping of the HPG2 region revealed the presence of sequence type (ST)1, ST4, and ST11, of which the latter has also been identified in Europe. The MIC susceptibility test showed that all tested isolates were susceptible for the majority of tested antimicrobials, including erythromycin and tetracycline, which are important antibiotics for the treatment of IC. The IC situation in Sonora State, Mexico, is complex because of the presence of serotypes A, B, and C. This finding emphasizes the importance of biosecurity in combination with the application of the most optimal vaccination programs in the control of IC in Sonora State, Mexico.

Effects of acute stressors experienced by five strains of layer breeders on measures of stress and fear in their offspring

Stressors experienced by layer breeders during egg production can lead to changes in the egg hormone content, potentially impacting their offspring, the commercial layers. Genetic differences might also affect the offspring's susceptibility to maternal experiences. In this study, we tested if maternal stress affects measures of stress and fear in five strains of layer breeders: commercial brown 1 & 2, commercial white 1 & 2 and a pure line White Leghorn. Each strain was equally separated into two groups: “Maternal Stress” (MS), where hens were subjected to a series of 8 consecutive days of acute psychological stressors, and “Control,” which received routine husbandry. Additional eggs from Control were injected either with corticosterone diluted in a vehicle solution (“CORT”) or just “Vehicle.” Stress- and fear-responses of the offspring were measured in a plasma corticosterone test and a combined human approach and novel object test. While the stress treatments did not affect the measured endpoints in the offspring, significant strain differences were found. The offspring of the white strains showed a higher physiological response compared to brown strains and the White 2 offspring was the least fearful strain in the human approach test. Our study found that neither the acute psychological stressors experienced by layer breeders nor the egg injections of corticosterone affected the parameters tested in their offspring. Post hoc power analyses suggest that the lack of treatment effects might be due to a small sample size (type II error). Although studies on larger flocks of layers are still needed, our results provide an initial understanding of an important subject, as in poultry production, layer breeders are often subjected to short-term stressors. In addition, our results suggest the dissociation between the physiological and behavioural parameters of stress response in laying hens, showing that increased concentrations of plasma corticosterone in response to stress might not be directly associated with high levels of fear.

Variation and Association of Hen Performance and Egg Quality Traits in Individual Early-Laying ISA Brown Hens.

Simple SummaryThe feed consumption and feed efficiency, body weight, egg weight, and rate of lay of the hen, as well as the albumen, yolk, and shell characteristics of the egg, are important measures of productivity in the layer industry. While these data are easily collected on a flock basis, there is little information on the extent of variation among individual hens in the literature and much nutrition and management is based on accepting the expected average performance of a cohort of animals. Moreover, body weight or feed consumption may have implications for other traits such as egg or egg component variables. We have attempted to control environmental variables such as feed in order to investigate the inherent variation for these traits present in a flock of 450 individually caged ISA Brown hens in early lay, and their associations with egg measurements. The results from the study revealed important variation in daily feed intake, body weight, and albumen height. Feed intake and hen body weights were weakly associated with egg weight, while feed intake and body weights were not associated with albumen height. Body weight and feed consumption had only a weak linkage with egg weight and did not influence albumen quality.Uniformity in hen and egg traits is an important consideration in commercial layer flocks. There is little information on how individual hen feed consumption and body weight affect egg quality measurements. This study investigated the variation in performance traits of individual hens and associations with egg quality characteristics. Four hundred and fifty-five ISA Brown caged hens in early lay were monitored for 42 days (25 to 30 weeks of age) to collect hen feed consumption and egg production measurements. Forty-four hens from the flock were randomly selected and eggs were collected from the same hen once weekly for albumen, yolk, and shell assessment. The means ± standard deviation of average daily feed intake (ADFI), albumen height, initial body weight (IBW), and final body weight (FBW) were 124 g ± 15, 10.3 mm ± 1.5, 1802 g ± 129, and 2000 g ± 175, respectively. Albumen height was not associated with ADFI (r = 0.18, p = 0.21), IBW (r = −0.04, p = 0.79), or FBW (r = −0.06, p = 0.69). This study showed variation in feed intake, body weight, and albumen quality of individual early-lay hens. Feed intake and body weight did not influence albumen quality.

English

Avian metapneumovirus (aMPV) is an important poultry pathogen causing an acute highly contagious upper respiratory tract infection in chickens, and could result in swollen head syndrome. The disease causes significant economic losses in turkey and chicken flocks; however, its existence in the Nigerian poultry industry is not well defined. Also, the influence of weather on the prevalence of the disease is unknown. This study was therefore carried out to determine the prevalence of Avian Metapneumovirus in chicken flocks from different climatic zones of Nigeria during both dry and wet seasons. Eighty commercial chickens (growers and adults) including layer and broiler flock from randomly selected local government areas from each of Sokoto, Plateau and Oyo States of Nigeria, were bled via jugular venipuncture for detection and titration of aMPV antibodies in serum “(n=240 serum samples each from both dry and wet season)”. Assay was carried out using a commercial enzyme-linked immunosorbent assay kit (ID Screen®, Rue Louis Pasteur-Grabels-France) which was able to determine antibodies against A, B, C and D subtypes of the virus.  Out of a total of 480 samples collected, 287 (59.8%) were positive for aMPV antibodies. During the dry season, prevalence was 100, 56.2 and 48.8% in Plateau, Sokoto and Oyo States, respectively, while it was 52.5, 65.2 and 36.2%, respectively during the wet season. Commercial chickens screened during the dry season (68.3%) and mean aMPV antibody titer of 2990.95 had significantly (α<0.001) higher seroprevalence of aMPV antibody compared to those screened in the wet season (51.2%) and mean aMPV antibody titer of 572.98. This study therefore shows that aMPV circulates more in the dry season in the studied areas of Nigeria. Key words: ELISA, avian metapneumovirus (aMPV), antibodies, weather conditions, seroprevalence, commercial chickens. &nbsp

An observational field study of the cloacal microbiota in adult laying hens with and without access to an outdoor range

BackgroundLaying hens with access to outdoor ranges are exposed to additional environmental factors and microorganisms, including potential pathogens. Differences in composition of the cloacal microbial community between indoor- and outdoor-housed layers may serve as an indicator for exposure to the outdoor environment, including its pathogens, and may yield insights into factors affecting the chickens’ microbiota community dynamics. However, little is known about the influence of outdoor housing on microbiota community composition in commercial layer flocks. We performed a cross-sectional field study to evaluate differences in the cloacal microbiota of indoor- vs outdoor-layers across farms.Eight layer flocks (four indoor, four outdoor) from five commercial poultry farms were sampled. Indoor and outdoor flocks with the same rearing flock of origin, age, and breed were selected. In each flock, cloacal swabs were taken from ten layers, and microbiota were analysed with 16S rRNA gene amplicon sequencing.ResultsHousing type (indoor vs outdoor), rearing farm, farm and poultry house within the farm all significantly contributed to bacterial community composition. Poultry house explained most of the variation (20.9%), while housing type only explained 0.2% of the variation in community composition. Bacterial diversity was higher in indoor-layers than in outdoor-layers, and indoor-layers also had more variation in their bacterial community composition. No phyla or genera were found to be differentially abundant between indoor and outdoor poultry houses. One amplicon sequence variant was exclusively present in outdoor-layers across all outdoor poultry houses, and was identified as Dietzia maris.ConclusionsThis study shows that exposure to an outdoor environment is responsible for a relatively small proportion of the community variation in the microbiota of layers. The poultry house, farm, and rearing flock play a much greater role in determining the cloacal microbiota composition of adult laying hens. Overall, measuring differences in cloacal microbiota of layers as an indicator for the level of exposure to potential pathogens and biosecurity seems of limited practical use. To gain more insight into environmental drivers of the gut microbiota, future research should aim at investigating community composition of commercial layer flocks over time.

Investigation on eggshell apex abnormality (EAA) syndrome in France: isolation of Mycoplasma synoviae is frequently associated with Mycoplasma pullorum

BackgroundMycoplasma synoviae (MS) is known to cause Eggshell Apex Abnormality (EAA) syndrome characterized by an altered shell surface with increased translucency on the apex. However, no large-scale studies have been conducted to obtain prevalence data of EAA and MS isolates associated to this syndrome. This manuscript reports the results of two field studies performed in the French poultry industry (2015–2017): focusing mainly on investigation of presence and prevalence of EAA in different types of laying hen flocks (phase 1), and isolation of MS strains from EAA-infected flocks (phase 2).ResultsThe first survey included 77 farms of commercial layers in three French egg-production regions, hosting 40 flocks in alternative systems (ALT) and 56 in furnished cages (FC). Seven flocks (4 FC and 3 ALT) presented EAA clinical signs, giving a prevalence of 7.3% in this studied sample. A second independent field study was conducted to identify MS by in vitro cultivation and PCR in samples from 28 flocks with clinical signs of EAA. Different types of biological specimens were collected in EAA-affected flocks and submitted to the laboratory. M. synoviae was detected in 25/28 flocks, from both production systems (5/5 ALT and 20/23 FC). Detection of MS was significantly higher in tracheal swabs (59%) than in cloacal (10.5%), albumen (3.6%) and egg yolk (1.1%) swabs. It is worth to mention that attempts to clone MS from positive samples were often hampered by the presence of another Mycoplasma species, which showed fast growing behaviour in the selective media used in this study (Frey Medium 4 and Frey Medium 4 supplemented with erythromycin). The use of MALDI-TOF mass spectrometry in combination with next-generation sequencing (NGS) results allowed the identification of this fast growing mycoplasma as Mycoplasma pullorum, which was detected in 14 of the 25 (56%) MS-positive flocks.ConclusionsThese results confirmed the presence of the EAA syndrome in MS-positive flocks of layers in France, reared in different regions and in different production systems (ALT and FC). Studies need to be conducted to test whether M. pullorum may influence the expression of clinical signs of EAA in MS-infected layer farms.

Welfare Assessment of Two Free-range Laying Hen Flocks in Turkey

ABSTRACT This study aimed to determine the prevalence of keel bone deviations and feather damage of laying hens in two different free-range housing systems under commercial conditions. Both of the free-range systems had an indoor barn and an outdoor range area. The floor of the indoor barn consisted of litter and either perches (litter and perch, LP) or slats (litter and slats, LS). The hens of both flocks were raised under identical conditions in the same house during the rearing period and then divided and transferred to two different free-range housing systems for the laying period. Examinations were conducted on the flocks at the end of the production cycle. Hens from the LP group had a greater prevalence of keel bone deviation and feather damage compared to hens from the LS group. The highest percentage of severe feather damage was found in the back and tail body regions in both flocks. The findings of this study are based upon the evaluation of two Turkish laying flocks. Therefore further research with more replicates of these treatments is needed to reach a general conclusion.

Gut microbiota profiles of commercial laying hens infected with tumorigenic viruses

BackgroundStudies have shown that some viral infections cause structural changes in the intestinal microflora, but little is known about the effects of tumorigenic viral infection on the intestinal microflora of chickens.ResultsA 29-week commercial layer flock positive for avian leukosis virus-J (ALV-J), Marek’s disease virus (MDV) and avian reticuloendotheliosis virus (REV) was selected, and fresh fecal samples were collected and examined for the composition of the gut microflora by Illumina sequencing of the V3-V4 region of the 16S rRNA gene. The operational taxonomic units (OTUs) of the fecal microbiota differentiated the chickens infected with only ALV-J and those coinfected with ALV-J and MDV or REV from infection-negative chickens. The enrichment and diversity of cloacal microflora in chickens infected with ALV-J alone were slightly different from those in the infection-negative chickens. However, the diversity of cloacal microflora was significantly increased in chickens coinfected with both ALV-J and MDV or REV.ConclusionsThe intestinal microbiota was more strongly disturbed in chickens after coinfection with ALV-J and MDV or REV than after infection with ALV-J alone, and there may be underlying mechanisms by which the capacity for the stabilization of the intestinal flora was impaired due to viral infection and tumorigenesis.

Evaluation of inactivated vaccine of the variant 2 (IS-1494 /GI-23) genotype of avian infectious bronchitis

The infectious bronchitis virus (IBV) is the cause of avian infectious bronchitis (IB). IB is one of the most highly contagious diseases, which results in many economic losses in the poultry industry worldwide. The nature of this virus is such that it generates new genotypes continuously. Proper vaccination is the most suitable way of combatting IB. One of the novel genotypes of IBV, which has been circulating in the Middle Eastern countries, is the variant 2 (IS-1494/GI-23) genotype. This study aims to design and produce an autogenous variant 2 vaccines. After isolation and characterization of the Iranian variant 2, the inactivated vaccine was formulated according to the OIE guidelines, and its different aspects (Purity, titration, inactivation, immunization) were evaluated. The designed vaccine passed all of OIE quality control standards. In the assessment process, the protection rate in the groups receiving the variant 2 and commercial vaccines was 67 % and 60 %, respectively. Although the differences were not significant, they indicated better protection, and the viral load in the feces and the kidney of the group receiving the variant 2 vaccine was lower than that in the commercial vaccine. It is suggested that the variant2 strain should be added as one of the local strains to the commercial inactivated vaccines in areas affected by this genotype. The use of this vaccine in layer and breeder flocks can help to protect them against variant 2 during the production phase. Also, the transfer of maternal antibodies to offspring can provide strain-specific immunity for one-day-old chickens.

Molecular characterisation and phylogenetic analysis of Marek’s disease virus in Turkish layer chickens

ABSTRACT 1. There is no current data about the genotypes of Marek’s disease virus (MDV) in Turkish poultry flocks; hence, this study was performed to analyse CVI988/Rispens, turkey herpesvirus (HVT) vaccine viruses and MDV field viruses as well as to perform phylogenetic analysis of MDV in Turkish layer chickens. 2. In 2017 and 2018, a total of 602 spleen samples from 49 layer flocks were collected from the Marmara, West Black Sea and Aegean regions. DNA was extracted from the spleen samples and the samples were analysed by real-time PCR probe assay to detect CVI988/Rispens and HVT vaccine viruses and MDV field strains. Samples found positive for MDV by real-time PCR were subjected to PCR using the Meq gene primers for phylogenetic analysis. 3. Amongst 49 flocks, virulent MDV was detected in nine flocks. CVI988/Rispens and HVT vaccine strains were detected in 47 flocks and HVT in all 49 flocks. Splenomegaly, hepatomegaly and tumours in the oviduct were observed in chickens of affected flocks. Virulent MDV was detected in 120 out of 602 spleen samples. Sequencing and phylogenetic analyses showed that MDVs detected in this study were closely related to MDV strains from Italy, Poland, Saudi Arabia, Iraq, India and China but showed diversity with MDV strains from Egypt and Hungary. Multiple sequence analysis of the Meq protein revealed several point mutations in deduced amino acid sequences. Interestingly, CVI988/Rispens vaccine virus from China (AF493555) showed mutations at position 66 (G66R) and 71 (S66A) along with two other vaccine strains from China (GU354326.1) and Russia (EU032468.1), in comparison with the other vaccine strain CVI988/Rispens (DQ534538). The molecular analyses of the Meq gene suggested that Turkish field strains of MDV are in the class of virulent or very virulent pathotypes. 4. The results have shown that MDV still affects poultry health, and the phylogenetic and amino acid variation data obtained will help in vaccination and control strategies.

Detection of Low Pathogenic Avian Influenza Virus Subtype H10N7 in Poultry and Environmental Water Samples During a Clinical Outbreak in Commercial Free-Range Layers, Netherlands 2017.

Wild birds are the natural reservoir of the avian influenza virus (AIV) and may transmit AIV to poultry via direct contact or indirectly through the environment. In the Netherlands, a clinically suspected free-range layer flock was reported to the veterinary authorities by the farmer. Increased mortality, a decreased feed intake, and a drop in egg production were observed. Subsequently, an infection with low pathogenic avian influenza virus was detected. This study describes the diagnostic procedures used for detection and subtyping of the virus. In addition to routine diagnostics, the potential of two different environmental diagnostic methods was investigated for detecting AIV in surface water. AIV was first detected using rRT-PCR and isolated from tracheal and cloacal swabs collected from the hens. The virus was subtyped as H10N7. Antibodies against the virus were detected in 28 of the 31 sera tested. An intravenous pathogenicity index (IVPI) experiment was performed, but no clinical signs (IVPI = 0) were observed. Post-mortem examination and histology confirmed the AIV infection. Multiple water samples were collected longitudinally from the free-range area and waterway near the farm. Both environmental diagnostic methods allowed the detection of the H10N7 virus, demonstrating the potential of these methods in detection of AIV. The described methods could be a useful additional procedure for AIV surveillance in water-rich areas with large concentrations of wild birds or in areas around poultry farms. In addition, these methods could be used as a tool to test if the environment or free-range area is virus-free again, at the end of an AIV epidemic.

Pathogenicity and Molecular Typing of Fowl Adenovirus-Associated With Hepatitis/Hydropericardium Syndrome in Central China (2015–2018)

In central China, a large number of broiler and layer flocks have suffered from outbreaks of severe hepatitis/hydropericardium syndrome (HHS). This resulted in huge economic losses to the poultry industry, from 2015 to 2018. To identify the specific pathogen and study its pathogenicity, 195 samples from Hubei, Jiangxi, Anhui, Hunan, and Henan provinces in central China were collected. The samples were screened for the adenovirus hexon gene, and neighbor joining was used for the phylogenetic reconstruction of the sequences. Among the collected samples, 122 were found to be positive for fowl adenovirus (FAdV) by PCR, and 73 isolates were obtained. The predominant viral serotype was serotype 4 (FAdV-4), which was found in 48 isolates, while 24 were serotype 10 (FAdV-10), and one was serotype 2 (FAdV-2). The CH/HBTF /1710 isolate was selected for further experiment and inoculated into 33-day-old specific pathogen-free chickens via intramuscular injection or oral administration to evaluate pathogenicity. It was found that the mortality for chickens infected by intramuscular injection or oral administration was 70 and 60%, respectively. Necropsy revealed mild to severe hepatitis and hydropericardium at 5 and 7 days after infection. Ancestor analyses indicated that all of the FAdV-4 strains obtained in this study shared a common Indian precursor and had a close genetic relationship with the JSJ13, SDSX, HN/151025, and SDDM-15 strains common in China.

The first complete genome sequence and pathogenicity characterization of fowl adenovirus 11 from chickens with inclusion body hepatitis in Pakistan

Inclusion body hepatitis (IBH), hydropericardium syndrome, and gizzard erosion associated with fowl adenovirus (FAdV) infections are reported globally and resulted in significant poultry industry economic losses. In 2018, severe IBH appeared in Pakistan in a 17-week-old layer flock. Subsequently, a FAdV-11 strain (designated as PKFAd18) was isolated from liver samples and identified based on phylogenetic analyses of the serotype-specific L1 region of the capsid hexon gene. There is no complete genome sequence of the Pakistani FAdV-11. This study successfully sequenced the complete genome of PKFAd18. The full genome of PKFAd18 contains 43 840 base pairs (bp) with a G + C content of 53.9 %, which is comparable to other FAdV serotypes. Similar to other FAdV-11 strains, PKFAd18 has only one fiber, while FAdV-1 and FAdV-4 have two fibers. Notably, PKFAd18 showed unique characteristics compared to other FAdV-11 strains. A natural large genomic deletion (1215 bp) appeared in tandem repeat region two, relative to the ON-NP2 strain. Phylogenetic analyses of the PKFAd18 penton gene showed higher homology with FAdV-9, highlighting potential natural recombination between FAdV-11 and FAdV-9. Moreover, the pathogenicity of PKFAd18 studied in specific-pathogen-free chickens showed that PKFAd18 is capable of inducing severe IBH and could be responsible for IBH in Pakistan. Thus, the first complete genome of FAdV-11 in Pakistan was sequenced in this study, which enriches the diversity of knowledge about FAdV-11 and is useful for developing diagnostics and vaccines for IBH induced by FAdV-11 in Pakistan.

Research Note: Molecular and pathologic characterization of avian adenovirus isolated from the oviducts of laying hens in eastern Japan

Cases of poor egg production were investigated in 2 layer farms from Ibaraki Prefecture in eastern Japan. To identify any microbial agents that may have caused the problem, necropsy, bacterial isolation, histopathology, and virus detection were performed. Members of the avian adenoviruses was detected by PCR in oviduct samples from both farms; chicken anemia virus coinfection was also confirmed in one of the farms. Avian adenovirus was isolated from the oviducts of the affected chickens on each farm. Inoculation into chick embryos showed tropism for the chorio-allantoic membrane. Stunting and hemorrhaging was observed in all infected embryos, as well as death in a few. Inoculation of 1-day-old specific pathogen-free chicks, and 400-day-old commercial hens, did not result in any significant findings. The isolated viruses were analyzed by sequencing of the hexon gene and were confirmed as fowl adenovirus type-c serotype-4 (FAdV-4). The 2 virus strains were found to be 99.29% similar to each other. One of the strains, Japan/Ibaraki/Y-H6/2016, was 99.15% similar to the KR5 strain. The other, Japan/Ibaraki/M-HB2/2016, was 99.57% similar to the KR5 strain. Fiber-2 gene analysis confirmed the identity as FAdV-4 that is closely related to nonpathogenic strains. Although nonpathogenic to chicks and laying hens, this infection can possibly cause economic damage. Perhaps the bigger concern is the effect on infected breeder operations. Because the virus is fatal to 9.09% of infected embryos, this could translate to a considerable loss in chick production owing to embryonic death. This is the first report of detection and isolation of FAdV-4 from the chicken oviduct; however, further studies are needed to elucidate its impact on both layer and breeder flocks. Indeed, FAdV-4 has negative effects on the avian reproductive tract as well.

Impacts and characteristics of antimicrobial resistance of Escherichia coli isolates by administration of third-generation cephalosporins in layer hatcheries

We investigated the characteristics and persistence of Escherichia coli resistant to third-generation cephalosporins (3GCs) by early administration of ceftiofur or gentamicin and to analyze the impact of 3GC use in hatcheries. We studied 10 ceftiofur-treated flocks (CTFs) and 10 gentamicin-treated flocks (GTFs) of layers. Fecal samples were collected at 1, 2, 4, 8, 18, and 30 weeks of age for all flocks. Among the 446 E. coli isolates, 58 (29.0 %) of 200 isolates in CTFs were identified as 3GC-resistant E. coli and 28 (11.4 %) of 246 isolates in GTFs were identified as 3GC-resistant E. coli. The presence of 3GC-resistant E. coli isolates at 1, 2, and 4 weeks was significantly higher in CTFs than in GTFs (p < 0.05). Moreover, the rate of resistance to 3GCs gradually decreased from 83.3 % at 1 week of age to 4.4 % at 30 weeks of age in CTFs. Of the 86 3GC-resistant E. coli isolates, 32 isolates had β-lactamase-encoding gene: blaCTX-M-14 (ten isolates), blaCTX-M-15 (three isolates), blaCMY-2 (five isolates), and blaTEM-1 (twenty-five isolates) genes. Plasmid replicon typing revealed that blaCTX-M-14, blaCTX-M-15, blaCMY-2, and blaTEM-1 were located on F, F and FIB, I1 and K, and I1 and FII, respectively. Furthermore, 18 isolates carried class 1 integrons, with four different gene cassettes. These results revealed that ceftiofur used in hatcheries can lead to an increase in the number of 3GC-resistant E. coli with many characteristics. A voluntary ban must be imposed on the use of 3GCs for 1-day-old chicks in poultry industry.

Isolation, Serological and Molecular Detection of Infectious Laryngotracheitis Virus (ILTV) in Chickens in Libya

Infectious Laryngotracheitis (ILT) is a viral respiratory disease of poultry caused by Gallid herpesvirus 1 leading to severe economic losses. The aim of the current study was to isolate and identify ILT virus in suspected outbreaks of one broiler flock (farm 1) and three layer flocks (farms 2,3 and 4) located in Tripoli. Samples of trachea were collected and preserved in transport media for viral isolation and some were preserved in 10% neutral formalin for histopathology. Broiler and Layer flocks were serologically monitored and tested for the presence of antibodies by ELISA against ILTV at first days old and later at slaughter age for broilers and before the onset of production for layers in which total of 290 blood samples were collected from six poultry farms (3 broiler flocks and 3 layer flocks).

Genetic variability of the Avian leukosis virus subgroup J gp85 gene in layer flocks in Lower Egypt.

Aim:This study aimed to determine the prevalence of layer flock tumor disease in Lower Egypt during the period of 2018-2019 and to undertake molecular characterization and determine the genetic diversity of all identified viruses.Materials and Methods:Forty samples were collected from layer chicken located in six governorates of Lower Egypt during the period of 2018-2019. Samples were taken from tumors in different organs. Tumor tissues were identified by histopathological sectioning and then further confirmed by a reverse-transcription polymerase chain reaction. Finally, genetic evolution of Avian leukosis virus (ALV-J) gp85 gene was studied.Results:All the study samples were negative for Marek’s disease virus, reticuloendotheliosis virus, ALV (A,B,C and D) and 20 samples were positive for ALV-J in backyard in six governrates. Sequencing of ALV-J gp85 gene was performed for six representative samples (one from each governorate), and they were found to be genetically related to prototype virus HPRS-1003 (identity percentage: 91.2-91.8%), but they were from a different group that was similar to the AF88-USA strain (first detected in 2000) with specific mutations, and they differed from a strain that was previously isolated in Egypt in 2005, forming two different subgroups (I and II) that had mutations in the hr1domain (V128F, R136A) and hr2 domain (S197G, E202K).Conclusion:The ALV-J virus was the main cause of neoplastic disease in layer chickens from Lower Egypt in the period of 2018-2019. We found that the genetic evolution of ALV-J gp85 gene was related to prototype virus HPRS-1003 but in a different group with a specific mutation. Further studies are needed to evaluate the antigenicity and pathogenicity of recently detected ALV-J strains.

First report on the molecular detection, phylogeny, virological and pathological investigations of Avibacterium paragallinarum in chickens in Turkey

Avibacterium paragallinarum is an important pathogen affecting the respiratory tract of chickens. There is a paucity of information on the molecular characteristics and pathology of A. paragallinarum in Turkish poultry flocks. In the present study, broiler and layer flocks (n = 2) suspected of viral infections with serious respiratory signs and significant mortality were visited. Chickens showed various disease signs and necropsy lesions, including purulent nasal discharge, respiratory distress, facial edema, sticky eyes, mucoid tracheitis, hemorrhagic inflammation of the infraorbital sinuses along with fibrinous mass and conjunctivitis. Histopathological lesions included loss of cilia along with necrosis and exfoliation of the superficial mucosal epithelium of the trachea, facial cellulitis, dermatitis, fibrinous plasmatic edema and infiltration of inflammatory cells, especially heterophils. A. paragallinarum was detected in tissue samples by species-specific polymerase chain reaction (PCR). The sequencing and phylogenetic analysis of the core region of the hemagglutinin (HA) gene revealed that Turkish strains detected here belonged to serotype A (serovar A1). They were related to strains reported from India (VRDC), the United States (0083), and Japan (0221), which are representatives of serovar A1. A homology of 88-90% was found between Indian strains and the Turkish strains detected in this study. Surprisingly, only vaccine strains of infectious bronchitis virus (IBV) were detected as a co-infecting agent in all samples positive for A. paragallinarum. Our findings suggest that A. paragallinarum may be an emerging pathogen in Turkish poultry flocks, and direct PCR may facilitate rapid diagnosis of infectious coryza. These results will also help to develop control strategies for A. paragallinarum.