Response of layer and broiler strain chickens to parenteral administration of a live Salmonella Typhimurium vaccine.

Responses to the parenteral administration of a live aroA deletion Salmonella serovar Typhimurium vaccine given to three brown egg layer strains and two broiler strains were studied. Twenty-five birds of each strain were reared together in floor pens to 6 weeks of age and then moved as individual strains to new floor pens and injected with 10(8) colony forming units (CFU) per bird of the vaccine bacteria intramuscularly or subcutaneously, 10(6) CFU per bird subcutaneously, or phosphate buffered saline (PBS) subcutaneously as a vaccination control. Three birds of one layer strain were injected intramuscularly with 0.5mg/ bird S. Typhimurium lipopolysaccharide (LPS) to evaluate whether response was similar for vaccine and endotoxin. Birds were weighed, and rectal temperatures recorded at the time of injection, then observed over 24 hours. Rectal temperatures were measured and blood samples collected for serum IL-6 assay at 3 hours post injection (PI). At 12 hours PI blood samples were drawn for analyses for plasma phosphorus (P), glucose (Glu), cholesterol (Cho), aspartate transaminase (AST), total protein (Ptn) and creatinine kinase (CK). Blood was sampled 14 days PI and tested for serum antibody to S. Typhimurium. Vaccination resulted in significant seroconversion by 14 days PI in all strains compared to the controls. The three layer strains exhibited a clinical malaise, evident within 90 minutes of injection, lasting for 12 hours, with complete recovery by 24 hours PI. Only the 10(8) CFU dose given subcutaneously produced an increase in rectal temperature 3 hours PI. Vaccination had no effect on IL-6 or Ptn. All vaccine doses increased P and the higher dose by either route decreased Cho in all bird strains. The 10(8) vaccine dose increased Glu and intramuscular injection markedly elevated CK only in the layer strains. The response was not completely congruous with that to LPS alone. The results highlight the need for consideration of differences in response of bird strain when consideration is given to the parenteral administration of live Salmonella vaccines.

Descriptive epidemiology of spotty liver disease in Australian cage-free brown egg layer chicken flocks with a scratch area.

Spotty Liver Disease (SLD), caused by Campylobacter hepaticus or C. bilis infection in adult female chickens continues to emerge as a major disease problem in cage-free production systems. Free range production has become the predominant system in Australian egg production and SLD is widespread in these farms. Previous studies have identified having a scratch area as a key determinant for SLD occurrence. An Australia-wide survey of egg production flocks with scratch areas was conducted regarding SLD including 48 individual flocks. Descriptive information on the facilities and flock management practices was reported. The incidence of SLD, age of first outbreak, initial mortality rate, duration of elevated mortality, and magnitude and duration of any associated egg production decline are described. Recurrence of SLD in the same flock was also reported and discussed. Therapies applied were recorded and assessed across SLD severity and duration. SLD occurred in 66.7% of layer flocks whose facility included a scratch area. Recurrent SLD outbreaks occurred in 31% of flocks experiencing SLD. Antibiotic medication reduced duration of mortality and egg production decline. Antibiotic therapy was associated with reduced duration of mortality and a less severe and shorter duration of egg production drops compared to untreated flocks. PCR detection of C. hepaticus in cloacal swabs and house dust samples and a serological ELISA test were compared and evaluated as diagnostic aids or as possible predictors of SLD outbreaks. The ELISA showed substantial agreement with detection of C. hepaticus in cloacal swabs by PCR. Examining composite house dust samples by PCR for C. hepaticus DNA appeared to be the most convenient and cost-effective aid to diagnosis and as a putative predictor for SLD outbreaks.

Development, application, and results of routine monitoring of Marek's disease virus in broiler house dust using real-time quantitative PCR.

Results are presented from four studies between 2002 and 2011 into the feasibility of routinely monitoring Marek's disease virus serotype 1 (MDV-1) in broiler house dust using real-time quantitative PCR (qPCR) measurement. Study 1 on two farms showed that detection of MDV-1 occurred earlier on average in dust samples tested using qPCR than standard PCR and in spleen samples from five birds per shed assayed for MDV-1 by qPCR or standard PCR. DNA quality following extraction from dust had no effect on detection of MDV-1. Study 2 demonstrated that herpesvirus of turkeys (HVT) and MDV serotype 2 (MDV-2) in addition to MDV-1 could be readily amplified from commercial farm dust samples, often in mixtures. MDV-2 was detected in 11 of 20 samples despite the absence of vaccination with this serotype. Study 3 investigated the reproducibility and sensitivity of the qPCR test and the presence of inhibitors in the samples. Samples extracted and amplified in triplicate showed a high level of reproducibility except at very low levels of virus near the limit of detection. Mixing of samples prior to extraction provided results consistent with the proportions in the mixture. Tests for inhibition showed that if the template contained DNA in the range 0.5-20 ng/microl no inhibition of the reaction was detectable. The sensitivity of the tests in terms of viral copy number (VCN) per milligram of dust was calculated to be in the range 24-600 VCN/mg for MDV-1, 48-1200 VCN/mg for MDV-2, and 182-4560 VCN/mg for HVT. In study 4 the results of 1976 commercial tests carried out for one company were analyzed. Overall 23.1% of samples were positive for MDV-1, 26.1% in unvaccinated and 16.4% in vaccinated chickens. There was marked regional and temporal variation in the proportion of positive samples and the MDV-1 load. The tests were useful in formulating Marek's disease vaccination strategies. The number of samples submitted has increased recently, as has the incidence of positive samples. These studies provide strong evidence that detection and quantitation of MDV-1, HVT, and MDV-2 in poultry house dust using qPCR is robust, sensitive, reproducible, and meaningful, both biologically and commercially. Tactical vaccination based on monitoring of MDV-1 rather than routine vaccination may reduce selection pressure for increased virulence in MDV-1.

Scratch area as an epidemiological risk factor for Spotty Liver Disease in cage-free layers in Australia.

Spotty Liver Disease (SLD) is a serious problem in laying hens farmed in cage-free systems. The causative organism, Campylobacter hepaticus, is regarded as having a fecal-oral method of transmission and hence may build up and spread readily in housing systems which allow ease of direct contact of hens with the flock's fecal material. The epidemiology of SLD has not been thoroughly investigated. An initial cross-sectional analytical epidemiological survey of SLD in free range and barn layer systems was conducted in Australia over 2019 to 2021.The survey involved rearing flocks (n = 32) which were then followed through into laying flocks (n = 24) up to 40 wk of age. Cloacal swabs were collected during rearing and lay for C. hepaticus detection by PCR. Flocks were classified as "Cases" (n = 18) where clinical SLD according to the case definition was observed or "Controls" (n = 6) which were clinically unaffected. No C. hepaticus was detected in cloacal swabs from rearing houses whereas the organism was detected in 18 Case flocks in lay and from 2 Control flocks in lay. All layer houses that incorporated a scratch area (n = 13) were categorized as Cases. Thus, having a scratch area is a key determinant for SLD and no analyses of further contributory factors from these flocks were able to be made. Of the remaining 11 flocks which had floors fully covered by slats, 5 were Cases (45%). Further risk factor analysis was compromised by this small sample size and identification of other significant associations was not possible. A larger survey investigating flocks laying in houses with fully slatted floors was undertaken to further the understanding of SLD epidemiology and is reported in a companion paper.

Identification of epidemiological risk factors for spotty liver disease in cage-free layer flocks in houses with fully slatted flooring in Australia.

Spotty liver disease (SLD) is recognized to be caused by infection with Campylobacter hepaticus in adult layer hens farmed in cage-free environments. SLD is an emerging disease as cage-free egg production increases in popularity in response to desires for improved welfare of poultry. Outbreaks of SLD are frequently experienced around peak egg production in flocks, commonly between 25 and 40 wk of age. The disease becomes manifest with increased exposure and access of the birds to the feces of the flock. This study follows from a previous epidemiological survey of free-range and barn flocks in Australia which identified the presence of a scratch area within the laying house as a major risk factor for the occurrence of SLD. However, that survey also observed SLD occurrence in 45% of houses with a fully slatted floor (no scratch area). The present study describes a further analytical survey aimed at identification of risk factors for SLD in houses with fully slatted flooring. A comprehensive questionnaire was completed for 49 cage-free flocks from point of lay until 40 wk of age across Australia, retrieving information on house design, bird breed, flock size, stocking densities, bird growth, and performance and the occurrence of SLD. Multiple logistic regression model building was used to separate factors and identify important management factors that may be amenable to modify the occurrence of SLD in egg layers. Key determinants of SLD identified from the analyses were that houses with mechanical ventilation (such as tunnel ventilation) have some protection from SLD and an increase of an extra 1 bird/m2 of nest space increased odds of occurrence of SLD by 1.172 times. A recommendation to not exceed 112 brown egg layer hens/m2 of nest space in naturally ventilated houses with a full slat floor was suggested. A delay in birds reaching 60% hen day production (HD) by 1 wk is suggested as a possible predictor for a subsequent outbreak of SLD.

Detection, characterization, and persistence of Campylobacter hepaticus, the cause of spotty liver disease in layer hens.

A Campylobacter species was first described as the etiological agent of Spotty Liver Disease (SLD) in 2015 and subsequently named as Campylobacter hepaticus in 2016. The bacterium predominantly affects barn and/or free-range hens at peak lay, is fastidious and difficult to isolate, which has impeded elucidation of its sources, means of persistence and transmission. Ten farms from South-Eastern Australia, of which 7 were free range entities participated in the study. A total of 1,404 specimens from layers and 201 from environmental sources, were examined for the presence of C. hepaticus. In this study, our principal findings included the continuing detection of C. hepaticus infection in a flock following an outbreak, indicating a possible transition of infected hens to asymptomatic carriers, that was also characterized by no further occurrence of SLD in the flock. We also report that the first outbreaks of SLD on newly commissioned free-range farms affected layers ranging from 23 to 74 wk of age, while subsequent outbreaks in replacement flocks on these farms occurred during the more conventional peak lay period (23-32 wk of age). Finally, we report that in the on-farm environment, C. hepaticus DNA was detected in layer feces, inert elements such as stormwater, mud, soil, as well as in fauna such as flies, red mites, Darkling beetles, and rats. While in off-farm locations, the bacterium was detected in feces from a variety of wild birds and a canine.

An evaluation of bird weight and diet nutrient density during early lay on ISA Brown performance, egg quality, bone characteristics, and liver health at 50 weeks of age.

This study compared the impact of a higher nutrient density (HND) or lower nutrient density (LND) diet fed during early lay to either heavier weight (HW) or lighter weight (LW) ISA Brown hens. At 18 wk of age (WOA) pullets (n = 240) were evenly assigned to either HW (n = 120) or LW (n = 120). Sixty birds from each weight group were then randomized to either the HND or LND diet treatments which were fed from 18 to 24 WOA inclusive. At 25 WOA the LND diet replaced the HND diet. All hens remained on LND diet to 50 WOA. Hen performance was measured from 18 to 50 WOA. Femur and liver health were evaluated at 50 WOA. Egg quality was assessed from 46 to 50 WOA. The 18 WOA HW hens had higher BW, cumulative egg production, cumulative feed intake (CFI), and cumulative egg mass (CEM) to both 24 and 50 WOA (P < 0.01). At 24 WOA the HND diet also generated higher BW (P < 0.001), CEM (P < 0.001) and lower cumulative feed conversion ratio (CFCR) (P < 0.01), the latter being sustained to 50 WOA (P < 0.01). At 50 WOA CFCR of LW birds was lower than HW birds (P < 0.01). Egg weight (EW), yolk diameter, and percent yolk weight were higher (P < 0.05) in the HW birds with the highest albumen to yolk ratio in LW birds (P < 0.05). Egg shape index was higher in LND diet fed birds (P < 0.01) while LW hens had higher shell phosphorus (P < 0.05). Body weight and diet nutrient density interacted on femoral diameter and cortical thickness being higher (P < 0.01) in LW birds fed HND than LW birds fed LND diets. Fatty liver hemorrhagic scores (P < 0.05) and liver lipid peroxidase (P < 0.001) at 50 WOA were higher in HW and LND diet treatments. Concurrently HW birds had the highest CFI and EW while CFCR and liver health were superior in LW and the HND diet treatment.

Seroprevalence of major respiratory diseases of chickens in central Ethiopia in different chicken production systems.

In Ethiopia, most chicken disease outbreaks and mortalities are attributed to a respiratory syndrome known as "fengil" with variable clinical signs and undefined etiology. The main goal of this study was to determine whether key respiratory pathogens that could contribute to the fengil syndrome circulate in Ethiopia. Specifically, we aimed to determine the seroprevalence of infectious laryngotracheitis virus (ILTV), infectious bronchitis virus (IBV), Newcastle disease virus (NDV), Mycoplasma gallisepticum (Mg), and avian metapneumovirus (aMPV). A cross-sectional survey was conducted in 158 scavenging and 42 small and medium-scale intensive chicken holdings in the East, West and North Shewa Zones of central Ethiopia. Blood from 495 chickens was collected and serological tests were used to determine exposure to these pathogens. Vaccination against NDV was the only immunization practiced with a significantly higher vaccination rate in the intensive than the scavenging system. Serological evidence of a high level of exposure to all pathogens was detected, including the first report on the seroprevalence of aMPV, ILTV, and IBV in the East Shewa Zone. The chicken and holding seroprevalence rates were respectively 91% and 94% for IBV, 34% and 57% for aMPV, 47% and 66% for Mg, 27% and 51% for ILTV and in unvaccinated flocks, 39% and 53% for NDV. These pathogens could contribute to the fengil syndrome, commonly ascribed to NDV. The seroprevalence of aMPV and ILTV was higher in chickens under the scavenging system. Exposure to multiple pathogens was common, with more than 50% of chickens positive for three or more pathogens in the scavenging system. This was reflected in significant positive associations between seropositivity to ILTV, Mg, ILTV, and IBV. The role of these pathogens in the causation of respiratory disease in the field requires further investigation.

Field Application of qPCR Monitoring of Infectious Laryngotracheitis Virus in Settled Chicken House Dust and Its Role in Control of a Major Outbreak.

Population-level sampling based on qPCR detection of infectious laryngotracheitis virus (ILTV) in poultry dust can be used to assess ILT vaccination outcomes following mass administration in drinking water. We report on the field application of this approach to assess the success of vaccine administration and its use in ILT outbreak control in meat chickens. In Study 1, dust samples were collected from 26 meat chicken flocks at 0, 4, 7, 14, and 21 days post drinking water vaccination (DPV) given between 7 to 13 days of age with the Serva or A20 live attenuated ILT vaccines. Unexpectedly, ILTV DNA was detected in dust samples collected prior to vaccination in 22/26 flocks. Typing revealed that the detected ILTV was different from the vaccine virus. To determine whether the detected ILTV DNA was from active infection or carryover of a noninfectious virus, Study 2 was implemented in 14 additional flocks with dust samples collected at 0, 7, 14, and 21 DPV and tracheal swabs collected from 15 birds/flock at 0 and 21 DPV. The results indicated that there was active infection with ILTV in those flocks before vaccination. This approach contributed to a statewide control program resulting in the eradication of ILT from South Australia as confirmed by negative ILTV test results for dust samples from 50 flocks and the absence of clinical ILT. These findings show that ILTV infection prior to vaccination is common in outbreak situations and that dust samples must be collected at 0 and 7 DPV for meaningful interpretation of vaccination outcomes and ILTV status. Comparatively low-cost dust testing during an outbreak, coupled with typing information, greatly assisted with decision making and control strategies during a major outbreak, including confirmation of the absence of infection in the final stages.

Aplicación de campo del monitoreo por qPCR del virus de la laringotraqueítis infecciosa en el polvo de casetas avícolas y su función en el control de un brote importante El muestreo a nivel de población basado en la detección por qPCR del virus de la laringotraqueítis infecciosa (ILTV) en el polvo de instalaciones avícolas se puede utilizar para evaluar los resultados de la vacunación contra esta enfermedad después de la administración masiva en el agua de bebida. Se reporta la aplicación de campo de este enfoque para evaluar el éxito de la administración de vacunas y su uso en el control de brotes por laringotraqueítis infecciosa en pollos de engorde. En el Estudio 1, se recolectaron muestras de polvo de 26 parvadas de pollos de engorda a los 0, 4, 7, 14 y 21 días después de la vacunación en el agua de bebida (DPV) a los 7 a 13 días de edad con las vacunas de laringotraqueítis vivas atenuadas Serva o A20. Inesperadamente, se detectó ADN del virus de laringotraqueítis en muestras de polvo recolectadas antes de la vacunación en 22/26 parvadas. La tipificación reveló que el virus detectado era diferente del virus de la vacuna. Para determinar si el ADN del virus de laringotraqueítis detectado procedía de una infección activa o del remanente de un virus no infeccioso, se implementó el Estudio 2 en 14 parvadas adicionales con muestras de polvo recolectadas a los 0, 7, 14 y 21 días después de la vacunación y de hisopos traqueales recolectados de 15 aves/parvada a los cero y 21 días después de la vacunación. Los resultados indicaron que había infección activa con el virus de laringotraqueítis en esas parvadas antes de la vacunación. Este enfoque contribuyó a un programa de control estatal que resultó en la erradicación de laringotraqueítis del sur de Australia, como lo confirmaron los resultados negativos de las pruebas del mismo virus para muestras de polvo de 50 parvadas y la ausencia de laringotraqueítis infecciosa clínica. Estos hallazgos muestran que la infección por el virus de la laringotraqueítis antes de la vacunación es común en situaciones de brotes y que las muestras de polvo deben recolectarse a los cero y 7 días después de la vacunación para una interpretación significativa de los resultados de la vacunación y el estado de esta enfermedad. Las pruebas de polvo comparativamente de bajo costo durante un brote, junto con la información de tipificación, ayudaron mucho con la toma de decisiones y con las estrategias de control durante un brote importante, incluida la confirmación de la ausencia de infección en las etapas finales.

Development and validation of a drag swab method using tampons and different diluents for the detection of members of Salmonella in broiler houses.

Members of the genus Salmonella represent a significant public health concern and also a colonizer of commercial poultry. Therefore, the early detection and management of colonized broiler breeders and their progeny is essential. There have been numerous methods for farm-based detection, with gauze-based drag swabs being the most commonly used. In the present study, the wet (boiled water, buffered peptone water and double-strength skin milk) tampon was compared with the gauze to determine the recovery rate (10(2) colony-forming units/swab) of five common poultry serovars of Salmonella and after cold (4°C/48 h) storage. The recovery was found to be equivalent when tested using the ISO6572:2002 method, for all diluents (Cohen's κ =1.0; sensitivity = 1.0; specificity = 1.0). The subsequent field trial (n = 15 farms) compared the tampon drag swab (TDS) with a statistically appropriate (90% confidence, detect 10% prevalence) number of faecal swabs (n = 22), which also showed high agreement between the TDS and faecal sampling (κ = 0.86; McNemar's χ(2) = 1.0; sensitivity = 0.9; specificity = 1.0). However, direct faecal sampling showed a wider diversity of serovars of Salmonella than the corresponding TDS. The TDS is a very sensitive, readily available and cost-effective screening method for salmonellas in broiler breeder houses. This TDS technique may be used for routinely screening of broiler houses, and faecal sampling would only be used to confirm colonization or contamination, and to measure flock serovar variance.

A molecular based method for rapid detection of Salmonella spp. in poultry dust samples.

Salmonellosis, caused by Salmonella spp., is a widely reported foodborne zoonosis frequently associated with ingestion of poultry products. Salmonella vaccination of chickens can be used to reduce bacterial shedding and risk of human infection. To determine Salmonella burden in chicken farms, culture methods of environmental samples that require a turn-around time of 5-7 days are usually used. Rapid screening using molecular assays such as PCR of pre-enriched broth has been reported for Salmonella spp. detection in feed, floor dust, and drag swabs within 2-3 days. Here we report an adaptation of the method for detection of Salmonella in poultry dust samples collected using a settle plate method under experimental conditions. Key features:•Passive dust sample collection using dry settle plates without media suspended from dropper lines of drinkers.•Small amount of sample required for the pre-enrichment process.•Quantification of Salmonella DNA with high sensitivity using an inexpensive extraction protocol.

Can a combination of vaccination, probiotic and organic acid treatment in layer hens protect against early life exposure to Salmonella Typhimurium and challenge at sexual maturity?

Day old layer chicks were challenged with Salmonella Typhimurium using a seeder bird technique. Treatment groups were untreated control, administration of a probiotic in drinking water weekly, vaccination by intramuscular injection of a live aro-A deletion mutant vaccine at 10 weeks of age (woa) followed by an oral dose at 16 woa, probiotic administration plus vaccination, vaccination plus the administration of an organic acid preparation in feed from 16 woa and a combination of probiotic, vaccine and organic acid. Faecal shedding was monitored by culture at 1, 2, 3, 4, 8, 12, 15, 17, 20, 21, 23 and 25 woa and in dust from settle plates by PCR at intervals from 8 woa. Birds from each group were separated at 17 and 18 woa and challenged orally with 106 CFU of S. Typhimurium. Both untreated and probiotic groups shed Salmonella until 56 days. Salmonella was also detected in dust from 8 until 12 woa but little after this. After vaccination, from sexual maturity (18 woa) all groups except those that were vaccinated with and without probiotic re-excreted Salmonella. The probiotic alone was ineffective against this re-excretion and all groups receiving organic acids shed Salmonella. At 17 woa, unchallenged controls were fully susceptible to caecal colonization, however all other groups showed reduced susceptibility, including the untreated challenged group. However, at 18 woa (sexual maturity) only the groups that were vaccinated with or without probiotic showed reduced susceptibility to colonization. The organic acid treated groups (including the vaccinated group) did not show a difference to the untreated controls. S. Typhimurium demonstrated an ability to re-emerge at sexual maturity, similar to other serovars. The vaccine assisted in limiting the re-excretion at sexual maturity and decreased susceptibility to subsequent challenge. Use of a probiotic augmented the vaccine's protective capacity.

Utilization of a novel autologous killed tri-vaccine (serogroups B [Typhimurium], C [Mbandaka] and E [Orion]) for Salmonella control in commercial poultry breeders.

An autologous killed trivalent vaccine (3x10(8) colony-forming units [CFU]), based on three Salmonella serovars (Typhimurium - serogroup B, Mbandaka - serogroup C, and Orion - serogroup E) prevalent in the flocks of Australian poultry companies, was developed using Salenvac techniques. At 20 weeks, hens vaccinated at 12 and 17 weeks as well as non-vaccinated hens were challenged (250 microl of 10(7) CFU) with autologous and heterologous serovars belonging to serogroup B (Typhimurium and Agona), serogroup C (Mbandaka and Infantis) and serogroup E (Orion and Zanzibar). Overall, vaccination resulted in a significant difference in carriage of Salmonella between non-vaccinated and vaccinated commercial Cobb hens (P <0.05) for serogroups B and C. However, due to low colonization rates in the non-vaccinated birds, no significant difference (P>0.05) could be determined for serogroup E. All vaccinated flocks produced a significant antibody response (P<0.001) to the S. Typhimurium vaccine strain, measured using a S. Typhimurium enzyme-linked immunosorbent assay (Guildhay), which peaked at 20 weeks of age, with 39% of the hens positive. Maternal antibodies were detected in 16% of the yolks from eggs produced by these flocks. There was a significant difference after challenge with Salmonella (P <0.05) among 1-day-old chicks from vaccinated versus non-vaccinated parents, when challenged using 10(4) CFU but not when challenged with 10(8) CFU. The success of this trial resulted in the incorporation of this vaccine into a Salmonella control system in commercial broiler breeder production.

Assessment of A20 infectious laryngotracheitis vaccine take in meat chickens using swab and dust samples following mass vaccination in drinking water.

Infectious laryngotracheitis, caused by the alphaherpesvirus infectious laryngotracheitis virus (ILTV), is an important disease of chickens. Partial control of this disease in meat chickens is commonly achieved by mass vaccination with live virus in drinking water. There is a need for a practical test to evaluate vaccination outcomes. For the Serva ILTV vaccine, quantitative real-time PCR (qPCR) enumeration of ILTV genome copies (GC) in flock level dust samples collected at 7-8 days post vaccination (dpv) can be used to differentiate flocks with poor and better vaccine take. This study aimed to validate this approach for A20, another widely used ILT vaccine in Australia. In four meat chicken flocks vaccinated with A20 in water using two different water stabilization times (20 or 40 min), swabs from the trachea and choanal cleft and dust samples were collected at 0, 7, 14 and 21 dpv. ILTV GC detection in swabs and dust was highest at 7 dpv and at this time ILTV GC load in dust was strongly and positively associated with vaccine take in individual birds assessed by swab samples. Choanal cleft swabs provided significantly fewer ILTV positive results than paired tracheal swab samples but the level of ILTV GC detected was similar. Water stabilization time had only minor effects on vaccination response in favour of the shorter time. Location of dust collection had no effect on viral load measured in dust samples. Dust samples collected at 0 and 7 dpv can be used to assess the vaccination status of flocks.

A practical method for assessing infectious laryngotracheitis vaccine take in broilers following mass administration in water: Spatial and temporal variation in viral genome content of poultry dust after vaccination.

Infectious laryngotracheitis is an important disease of chickens caused by infectious laryngotracheitis virus (ILTV). Outbreaks commonly occur in meat chicken flocks and mass vaccination with live attenuated vaccines, usually in water, is used to control the disease in these populations. Vaccination with live virus via water and nipple drinkers requires stringent adherence to protocols to ensure success, but vaccine administration monitoring is not currently assessed due to a lack of economically viable methods. Vaccinal ILTV has been shown to be detectable in dust in experimental studies and has potential as a method of assessing vaccination success. However, the pattern of vaccinal ILTV detection in dust following vaccination under commercial conditions has not been defined. We report the longitudinal profile of ILTV genome copies (GC) in poultry house dust collected on settle plates following vaccination of 8 flocks of commercial meat chickens on four farms. ILTV GC was enumerated using quantitative real-time polymerase chain reaction (qPCR). There was considerable variation between flocks in the levels of ILTV GC detected post vaccination and this variation was significantly associated with vaccine take measured in individual birds in a companion study. There was no effect of sampling location on ILTV GC in dust but the amount of dust collected was greater in locations closer to the exhaust fans in artificially ventilated houses. Results indicate that measurement of ILTV GC in single or pooled dust samples at 7-8 days post vaccination enables detection of poor vaccine takes and provides a practical means of monitoring ILT vaccination.

The intake pattern and feed preference of layer hens selected for high or low feed conversion ratio.

Feed accounts for the greatest proportion of egg production costs and there is substantial variation in feed to egg conversion ratio (FCR) efficiency between individual hens. Despite this understanding, there is a paucity of information regarding layer hen feeding behaviour, diet selection and its impact on feed efficiency. It was hypothesised that variation in feed to egg conversion efficiency between hens may be influenced by feeding behaviour. For this experiment, two 35-bird groups of ISA Brown layers were selected from 450 individually caged hens at 25-30 weeks of age for either low FCR < 1.8 ± 0.02 (high feed efficiency (HFE) or high FCR > 2.1 ± 0.02 (low feed efficiency (LFE)). For each of these 70 hens, intake of an ad-libitum mash diet at 2-minute time intervals, 24 h a day, for 7 days was determined alongside behavioural assessment and estimation of the selection of components of the mash. The group selected for HFE had a lower feed intake, similar egg mass and associated lower FCR when compared with the LFE group. Whilst feed intake patterns were similar between HFE and LFE hens, there was a distinct intake pattern for all layer hens with intake rate increasing from 0300 to 1700 h with a sharp decline to 2200 h. High feed efficiency hens selected a diet with 25% more ash and 4% less gross energy than LFE hens. The LFE hens also spent more time eating with more walking events, but less time spent resting, drinking, preening and cage pecking events as compared with HFE hens. In summary, there was no contrasting diurnal pattern of feed consumption behaviour between the groups ranked on feed efficiency, however high feed efficiency hens consumed less feed and selected a diet with greater ash content and lower gross energy as compared with LFE hens. Our work is now focused on individual hen diet selection from mash diets with an aim of formulating precision, targeted diets for greater feed efficiency.

Spatial and temporal variation of Marek's disease virus and infectious laryngotracheitis virus genome in dust samples following live vaccination of layer flocks.

Monitoring of Marek's disease virus (MDV) and infectious laryngotracheitis virus (ILTV) genome using poultry dust can be useful to monitor on-farm vaccination protocols but there are no set guidelines for collection of this sample type. This study assessed different dust collection methods for MDV and ILTV detection in a vaccinated layer flock (n = 1700) from day-old to 50 weeks of age. Birds were vaccinated against MDV at day-old, and ILTV by drinking water at week 6 and eye drop at week 12. Dust samples were collected weekly by settle plates (1-3 plates/15 m2) or by scraping surfaces in the poultry shed and tested for ILTV and MDV genomic copies (GC) by PCR. ILTV GC were detected 4 weeks post water vaccination, peaked at weeks 12-14 and became mostly undetectable after week 18. MDV was detected in dust on week 1, peaked at weeks 3-6, declined 3 logs by week 26 and remained detectable at this level until week 50. There was no difference in the detection rates of ILTV and MDV collected from settle plates in different locations of the shed (P > 0.10). There was no difference between settle plate and scraped samples in ILTV GC load but higher MDV GC were found in scraped samples. The settle plate method appears to reflect the current level of vaccine virus in the flock while the scrape method likely represents a cumulative record of shedding. Assessment of viral GC in dust samples is a good candidate for a practical method of estimating successful vaccine administration.

Uptake and spread of infectious laryngotracheitis vaccine virus within meat chicken flocks following drinking water vaccination.

Vaccination against infectious laryngotracheitis virus (ILTV) in commercial broiler flocks in the field, which is only undertaken in the face of a local outbreak, requires mass administration techniques, usually via drinking water. This is often fraught with difficulties such as variable vaccination "reactions" and sometimes, vaccination failure. Laboratory testing of the outbreak strains however invariably shows the vaccines in use to be protective. To investigate this paradox, the dynamics of an ILT vaccine virus was examined within broiler flocks during a natural outbreak. In an initial flock, 70 birds were individually identified and had tracheal swabs collected sequentially at intervals from 1 to 26 days after vaccination and submitted for ILTV detection using qPCR. This evaluation was extended by collection of tracheal swabs from 40 to 45 random birds at 4, 7-8, 12-13 and 25-26 days post vaccination (pv) across a further 7 flocks. The results showed a very variable early uptake of vaccine virus from the drinking water (between 3% and 52% of tested birds with detectable virus in trachea at 4 days pv) and revealed that actual vaccination of the flocks relied on bird to bird transmission of the vaccine virus. In flocks with very low (<10%) initial bird uptake, successful exposure of vaccine virus to the majority of the flock can be delayed, leaving a large proportion of birds as susceptible at the likely time of possible exposure to wild virus. This may explain the cases of apparent failure of vaccination in the field. The variable bird to bird spread can be associated with reversion to virulence, this may explain the rolling vaccine reactions often observed. The variation in initial vaccine uptake may be affected by some factors involved with the administration technique and this requires further study in a larger sample size.

Kinetics of Marek's disease virus (MDV) infection in broiler chickens 1: effect of varying vaccination to challenge interval on vaccinal protection and load of MDV and herpesvirus of turkey in the spleen and feather dander over time.

Two experiments in commercial broiler chickens vaccinated with herpesvirus of turkeys (HVT) and challenged with Marek's disease virus (MDV) investigated the effects of the vaccination-to-challenge interval (VCI) on vaccinal protection against Marek's disease, and the kinetics of MDV and HVT load in the spleen and feather dander determined using real-time quantitative polymerase chain reaction. Experiment 1 in isolators tested VCI of 2, 4 and 7 days, while Experiment 2 in floor pens tested VCI of 0, 2, 4, 7 and 10 days. MDV challenge induced gross Marek's disease lesions in 14% to 74% of chickens by 56 days post-challenge. Vaccinal protection increased from approximately 40% to approximately 80% with increasing VCI between days 2 and 7 in both experiments, but not thereafter. MDV was detected in both the spleen and dander at 7 days post-challenge and increased rapidly to approximately 21 days post-challenge, after which levels plateaued, rose or fell gradually depending on treatment. HVT was also shed in significant amounts, 1 to 2 logs lower than for MDV, with a clear peak around 14 to 21 days post-vaccination. Vaccination significantly reduced the log(10)MDV load in the spleen (vaccinated, 2.99+/-0.20/10(6) spleen cells; unvaccinated, 4.60+/-0.23/10(6) spleen cells) and dander (vaccinated, 5.28+/-0.13/mg; unvaccinated, 6.00+/-0.18/mg) from infected chickens. The MDV load had a significant negative association with the VCI and the level of vaccinal protection. Measurement of dander production in Experiment 1 and the dust content of air in Experiment 2, combined with determination of the MDV load in these, enabled estimation of total daily shedding rates of MDV per chicken and of the MDV load in air for the first time.

Ranging behavior relates to welfare indicators pre- and post-range access in commercial free-range broilers.

Little is known about the effect of accessing an outdoor range on chicken welfare. We tracked individual ranging behavior of 538 mixed-sex Ross 308 chickens on a commercial farm across 4 flocks in winter and summer. Before range access, at 17 to 19 d of age, and post-range access, at 30 to 33 and 42 to 46 d of age in winter and summer flocks respectively, welfare indicators were measured on chickens (pre-range: winter N = 292; summer N = 280; post-range: winter N = 131; summer N = 140), including weight, gait score, dermatitis and plumage condition. Post-ranging autopsies were performed (winter: N = 170; summer: N = 60) to assess breast burn, leg health, and ascites. Fewer chickens accessed the range in winter flocks (32.5%) than summer flocks (82.1%). Few relationships between welfare and ranging were identified in winter, likely due to minimal ranging and the earlier age of post-ranging data collection compared to summer flocks. In summer flocks prior to range access, chickens that accessed the range weighed 4.9% less (P = 0.03) than chickens that did not access the range. Pre-ranging weight, gait score, and overall plumage cover predicted the amount of range use by ranging chickens in summer flocks (P < 0.01), but it explained less than 5% of the variation, suggesting other factors are associated with ranging behavior. In summer flocks post-range access, ranging chickens weighed 12.8% less than non-ranging chickens (P < 0.001). More range visits were associated with lower weight (P < 0.01), improved gait scores (P = 0.02), greater breast plumage cover (P = 0.02), lower ascites index (P = 0.01), and less pericardial fluid (P = 0.04). More time spent on the range was associated with lower weight (P < 0.01) and better gait scores (P < 0.01). These results suggest that accessing an outdoor range in summer is partly related to changes in broiler chicken welfare. Further investigations are required to determine causation.