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.

Developing a selective culturing approach for Campylobacter hepaticus.

Campylobacter hepaticus, the causative agent of Spotty Liver Disease (SLD) is an important disease in cage-free egg producing chickens causing mortality and production drops. C. hepaticus is a slow growing Campylobacter easily overgrown by fecal bacteria. It is currently only reliably isolatable from bile samples. A selective media for isolation from feces or environment would assist diagnosis and impact assessment. Growth of five Australian C. hepaticus isolates was studied using Horse blood agar (HBA), sheep blood agar (SBA), Bolton, Preston and Brain Heart Infusion (BHI) base media. Blood and/or bile were added to Bolton, Preston and BHI medias. C. jejuni was used as a positive control. Plates were incubated in duplicate under microaerophilic conditions at 42°C for 10 days and examined at days 3-5 and 7-10 of incubation. Each isolate was examined for sensitivity to 14 antimicrobials using HBA sensitivity plates. Growth was inhibited by BHI and by added bile, while blood improved growth. Further replicates using SBA, HBA, Bolton and Preston media showed best growth on Bolton agar with blood. All five C. hepaticus isolates were resistant to trimethoprim and vancomycin, while four were also resistant to rifampicin and bacitracin. Media based upon Bolton plus blood supplemented with vancomycin and trimethoprim might be used as the most appropriate media for selective growth of C. hepaticus. The addition of bile to media for C. hepaticus isolation and growth will inhibit growth and is not advised.

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.

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.

Oral Inoculation of Point-of-Lay Hens with the New South Wales Outbreak Strain of Salmonella Enteritidis Phage Type 12 Causes Infection, but Minimal Histopathology.

An outbreak of food poisoning in New South Wales (NSW) Australia in 2018, caused by Salmonella enterica serovar Enteritidis phage type 12 (PT12), was traced to eggs consumed from a NSW layer flock. This was the first report of Salmonella Enteritidis infection in NSW layer flocks, despite ongoing environmental monitoring. Clinical signs and mortalities were minimal in most flocks, although seroconversion and infection were demonstrated in some flocks. An oral dose-response challenge study with Salmonella Enteritidis PT12 was undertaken in commercial point-of-lay hens. Cloacal swabs collected at 3, 7, 10, and 14 days postinoculation and caeca, liver, spleen, ovary, magnum, and isthmus tissues collected at necropsy at either 7 or 14 days were processed for Salmonella isolation (AS 5013.10-2009 from ISO6579:2002). Histopathology was performed on the above tissues, as well as lung, pancreas, kidney, heart, and additional intestinal and reproductive tract tissues. Salmonella Enteritidis was consistently detected in cloacal swabs between 7 and 14 days postchallenge. The Salmonella Enteritidis PT12 isolate successfully colonized the gastrointestinal tract, liver, and spleen of all hens orally challenged with 107, 108, and 109 Salmonella Enteritidis, and less consistently colonized their reproductive tracts. On histopathology, mild lymphoid hyperplasia in the liver and spleen, along with hepatitis, typhlitis, serositis, and salpingitis, was observed at 7 and 14 days postchallenge, with a greater proportion of affected birds in the two higher dose groups. Diarrhea and culture of Salmonella Enteritidis from heart blood were not detected in challenged layers. The NSW isolate of Salmonella Enteritidis PT12 was able to invade and colonize the birds' reproductive tracts as well as a wide range of other tissues, indicating the potential for these naive commercial hens to contaminate their eggs.

La inoculación oral de gallinas ponedoras en el pico de postura con la cepa de Salmonella Enteritidis PT12 del brote en Nueva Gales del Sur causa infección, pero una histopatología mínima. Un brote de intoxicación alimentaria en Nueva Gales del Sur (NSW), Australia en 2018, causado por Salmonella enterica serovar Enteritidis fagotipo 12, se rastreó hasta los huevos consumidos de una parvada de ponedoras de NSW. Este fue el primer informe de infección por Salmonella Enteritidis en parvadas de ponedoras de NSW, a pesar del monitoreo ambiental continuo. Los signos clínicos y la mortalidad fueron mínimos en la mayoría de las parvadas, aunque se demostró seroconversión e infección en algunas parvadas. Se llevó a cabo un estudio de desafío oral para evaluar la dosis y su respuesta para Salmonella Enteritidis PT12 en gallinas ponedoras comerciales. Los hisopos cloacales recolectados a los tres, siete, diez y 14 días posteriores a la inoculación y los tejidos de ciego, hígado, bazo, ovario, magnum e istmos recolectados en la necropsia a los siete o 14 días se procesaron para el aislamiento de Salmonella (AS 5013.10-2009 del estándar ISO6579: 2002). Se realizó histopatología en los tejidos anteriormente mencionados, así como de pulmón, páncreas, riñón, corazón y tejidos intestinales y del tracto reproductivo adicionales. Salmonella Enteritidis se detectó consistentemente en hisopos cloacales entre los siete y 14 días después del desafío. El aislado de Salmonella Enteritidis PT12 colonizó con éxito el tracto gastrointestinal, el hígado y el bazo de todas las gallinas desafiadas por vía oral con dosis de 107, 108 y 109 de Salmonella Enteritidis, pero colonizó de manera menos consistente sus tractos reproductivos. En la histopatología, se observó hiperplasia linfoide leve en el hígado y el bazo, junto con hepatitis, tiflitis, serositis y salpingitis, a los siete y 14 días posteriores a la exposición, con una mayor proporción de aves afectadas en los dos grupos de dosis más altas. En las ponedoras desafiadas no se detectaron diarrea ni cultivo de Salmonella Enteritidis de sangre colectada del corazón. El aislamiento de Salmonella Enteritidis PT 12 de Nueva Gales del Sur pudo invadir y colonizar los tractos reproductivos de las aves, así como una amplia gama de otros tejidos, lo que indica el potencial de estas gallinas comerciales sin inmunidad para contaminar sus huevos.

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.

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.