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.

In vivo label-free optical signatures of chemotherapy response in human pancreatic ductal adenocarcinoma patient-derived xenografts.

Pancreatic cancer is a devastating disease often detected at later stages, necessitating swift and effective chemotherapy treatment. However, chemoresistance is common and its mechanisms are poorly understood. Here, label-free multi-modal nonlinear optical microscopy was applied to study microstructural and functional features of pancreatic tumors in vivo to monitor inter- and intra-tumor heterogeneity and treatment response. Patient-derived xenografts with human pancreatic ductal adenocarcinoma were implanted into mice and characterized over five weeks of intraperitoneal chemotherapy (FIRINOX or Gem/NabP) with known responsiveness/resistance. Resistant and responsive tumors exhibited a similar initial metabolic response, but by week 5 the resistant tumor deviated significantly from the responsive tumor, indicating that a representative response may take up to five weeks to appear. This biphasic metabolic response in a chemoresistant tumor reveals the possibility of intra-tumor spatiotemporal heterogeneity of drug responsiveness. These results, though limited by small sample size, suggest the possibility for further work characterizing chemoresistance mechanisms using nonlinear optical microscopy.

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.

Effect of restricted feeding on hen performance, egg quality and organ characteristics of individual laying hens.

This study was to assess the impact of permanent or temporary restricted feeding on laying hen production traits, physiology, and egg quality. Two hundred and forty individually housed ISA Brown hens were monitored across 2 phases, assigned to 3 treatments: ad libitum feeding (ALF), temporary restricted feeding (TRF) and permanent restricted feeding (PRF), n = 80 hens per treatment. In Phase 1 (P1), 22 to 40 weeks, the TRF and PRF hens were offered 115 g of feed daily. In Phase 2 (P2), 41 to 46 weeks, the TRF hens were transitioned to ALF status while the ALF and PRF hens remained as in P1. From 35 to 40 weeks, eggs were collected once weekly from 15 hens per treatment and assessed for differences in albumen, yolk, and shell variables. At 45 weeks, 10 hens each from the ALF and PRF groups were euthanized and differences in organ characteristics were assessed. In P1, feed intake, feed to egg conversion ratio and body weight (BW) change were lower (P < 0.01), while albumen height and Haugh unit were higher (P < 0.01) in both PRF and TRF hen treatments compared to hens allocated the ALF treatment. In P2, TRF and ALF hens had a higher egg production and egg mass than PRF (P < 0.01) than ALF. Body weight change in P2 was higher in TRF and similar in both ALF and PRF, while feed intake and feed conversion ratio were higher in TRF followed by ALF and least in the PRF treatment group (P < 0.01). At 45 weeks ALF hens had a greater abdominal fat pad weight and fatty liver haemorrhagic syndrome lesion score compared to PRF. Restricting hens to 115 g of feed per day from point of lay restrained BW, improved feed conversion ratio and albumen quality and reduced abdominal fat pad deposition and clinical signs of fatty liver haemorrhagic syndrome in individually housed laying hens.

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.

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.

Association of feed efficiency with organ characteristics and fatty liver haemorrhagic syndrome in laying hens.

Poor feed efficiency (FE) in hens impacts body weight (BW) and may reflect suboptimal health. Fatty Liver Haemorrhagic Syndrome (FLHS) is mostly observed in laying hens and affects egg production and hen performance. The aim of this study was to investigate the relationships of FE and BW with organ characteristics, liver composition and incidence of FLHS of 150 individually housed ISA Brown hens ranked on the basis of feed conversion ratio (FCR) attained from early lay. At 45 weeks, 10 birds per FE group (HFE-High feed efficient; MFE-medium feed efficient; LFE-low feed efficient) were randomly selected and euthanized. Hen BW was positively associated with feed intake and FCR. The HFE hens had a lower abdominal fat pad and liver weight compared to LFE hens. FLHS lesion score was higher (worse) in the LFE than HFE hen group and was moderately positively associated with BW and abdominal fat pad, but strongly positively associated with liver weight. Liver pathology of LFE hens showed hepatocytes with abnormal retention of lipids causing distended cytoplasmic vacuoles compared to the HFE hens. Hens which exhibited poorer FE in early lay had heavier abdominal fat pads, heavier, fatter livers and were more prone to FLHS.

The role of hen body weight and diet nutrient density in an extended laying cycle.

The egg production (EP), egg quality and health of heavier or lighter hens fed a diet of either higher nutrient density (HND) or lower nutrient density (LND) during early lay, was assessed at very late lay. Based on their body weight (BW) at 18 wk of age (WOA) ISA Brown pullets were allocated as either heavier weight (HW; average 1.65 kg) or lighter weight (LW: average 1.49 kg). Half of each BW group received the HND (2,901 kcal/kg; 17.6% crude protein (CP) or LND (2726 kcal/kg, 16.4% CP) diet from 18 to 24 WOA. From 25 to 90 WOA all birds received identical early, then mid and late-lay diets. Hen BW was measured after peak-lay (36 WOA) and at 90 WOA. At 89 WOA and across 18 to 36 and 18 to 89 WOA feed intake (FI), EP, egg mass (EM), and feed conversion ratio (FCR) were calculated. Eggshell quality, breast score, relative ovary weight and liver and bone health were evaluated in very late lay. Differences in BW continued to 90 WOA. At 36 WOA HW hens produced heavier eggs, and had higher 18 to 36 WOA cumulative FI, EM (P < 0.001) and FCR (P < 0.05). When 89 WOA HW birds consumed more feed (P < 0.001) but EP, EM and FCR did not differ from LW hens. Cumulatively, 18 to 89 WOA FI and EM were higher for HW hens (P < 0.05), but cumulative EP and FCR was not different. The early-lay HND diet improved very late lay eggshell thickness (P < 0.05) and shell breaking strength (P = 0.05). Lighter hens fed HND and HW hens fed LND diet produced heavier eggs, higher relative oviduct weight and lower liver lipid peroxidase in very late lay (P < 0.05). Bone strength did not differ, but LW hens had higher femoral manganese and zinc (P < 0.05), lowering their likelihood of osteoporosis. Overall LW hens sustained EP throughout a longer laying cycle with beneficial bone characteristics. The HND diet improved eggshell strength and, in LW hens reduced hepatic oxidation.

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.

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.

The influence of hen size and diet nutrient density in early lay on hen performance, egg quality, and hen health in late lay.

The effect of hen size and diet nutrient density during early lay on egg production (EP) at 24 and 69 wk of age (WOA) and late lay egg quality and hen health was evaluated. Based on bodyweight (BW) at 18 WOA ISA Brown hens were assigned as heavier (HW; n = 120) or lighter weight (LW; n =120). Sixty birds from each BW group were fed an early-lay diet of higher nutrient density (HND), or lower nutrient density (LND) between 18 and 24 WOA. From 25 WOA all hens received the same early-lay diet and then from 40 WOA the mid-lay diet. Hen average daily feed intake (ADFI), hen-day EP, egg weight (EW), egg mass (EM), and feed conversion ratio (FCR) were assessed at 24 and 69 WOA. Between 66 and 70 WOA eggshell and internal egg quality was evaluated and at 70 WOA BW, liver and bone health were assessed. At 24 WOA BW was highest in HW birds and birds receiving the HND diet (P < 0.01). Concurrently ADFI, and FCR were higher and hen-day EP was lower in HW compared to LW birds (P < 0.05). The HND diet resulted in lower ADFI and FCR at 24 WOA, but higher EW and EM compared to the LND diet (P < 0.01). At 69 WOA HW birds had higher ADFI, EW (P < 0.02) and heavier 70 WOA BW compared to LW hens. The lower FCR of the LW birds at 69 WOA was approaching significance (P = 0.054). Hen weight and diet density did not affect 69 WOA egg production. Between 18 and 69 WOA cumulative FI and EM were higher in HW hens (P < 0.01) than LW hens, as was cumulative FCR (P = 0.053). Hen weight and diet density did not alter 66-70 WOA internal egg quality, but the HND diet generated thicker eggshells and higher eggshell breaking strength (P < 0.05). Seventy WOA liver health, keel curvature and femur breaking strength did not differ. Overall LW hens had lower FCR than HW hens and the early-lay HND diet facilitated improved eggshell integrity during late lay compared to the LND diet.

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.

Effectiveness of tests to detect the presence of SARS-CoV-2 virus, and antibodies to SARS-CoV-2, to inform COVID-19 diagnosis: a rapid systematic review.

OBJECTIVES: We undertook a rapid systematic review with the aim of identifying evidence that could be used to answer the following research questions: (1) What is the clinical effectiveness of tests that detect the presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) to inform COVID-19 diagnosis? (2) What is the clinical effectiveness of tests that detect the presence of antibodies to the SARS-CoV-2 virus to inform COVID-19 diagnosis? DESIGN AND SETTING: Systematic review and meta-analysis of studies of diagnostic test accuracy. We systematically searched for all published evidence on the effectiveness of tests for the presence of SARS-CoV-2 virus, or antibodies to SARS-CoV-2, up to 4 May 2020, and assessed relevant studies for risks of bias using the QUADAS-2 framework. MAIN OUTCOME MEASURES: Measures of diagnostic accuracy (sensitivity, specificity, positive/negative predictive value) were the main outcomes of interest. We also included studies that reported influence of testing on subsequent patient management, and that reported virus/antibody detection rates where these facilitated comparisons of testing in different settings, different populations or using different sampling methods. RESULTS: 38 studies on SARS-CoV-2 virus testing and 25 studies on SARS-CoV-2 antibody testing were identified. We identified high or unclear risks of bias in the majority of studies, most commonly as a result of unclear methods of patient selection and test conduct, or because of the use of a reference standard that may not definitively diagnose COVID-19. The majority were in hospital settings, in patients with confirmed or suspected COVID-19 infection. Pooled analysis of 16 studies (3818 patients) estimated a sensitivity of 87.8% (95% CI 81.5% to 92.2%) for an initial reverse-transcriptase PCR test. For antibody tests, 10 studies reported diagnostic accuracy outcomes: sensitivity ranged from 18.4% to 96.1% and specificity 88.9% to 100%. However, the lack of a true reference standard for SARS-CoV-2 diagnosis makes it challenging to assess the true diagnostic accuracy of these tests. Eighteen studies reporting different sampling methods suggest that for virus tests, the type of sample obtained/type of tissue sampled could influence test accuracy. Finally, we searched for, but did not identify, any evidence on how any test influences subsequent patient management. CONCLUSIONS: Evidence is rapidly emerging on the effectiveness of tests for COVID-19 diagnosis and management, but important uncertainties about their effectiveness and most appropriate application remain. Estimates of diagnostic accuracy should be interpreted bearing in mind the absence of a definitive reference standard to diagnose or rule out COVID-19 infection. More evidence is needed about the effectiveness of testing outside of hospital settings and in mild or asymptomatic cases. Implementation of public health strategies centred on COVID-19 testing provides opportunities to explore these important areas of research.

Hens That Exhibit Poorer Feed Efficiency Produce Eggs with Lower Albumen Quality and Are Prone to Being Overweight.

Feed efficiency (FE) is an important measure of productivity in the layer industry; however, little is known about how FE differs between individual hens during the egg-laying cycle and the implications for egg quality parameters. Individual 25-week-old ISA Brown hens were observed for 42 days, ranked into three FE groups (n = 48 per High (HFE), Medium (MFE) and Low (LFE) FE groups and then monitored later in the laying cycle from 35-40 weeks. The groups exhibited different feed to egg conversion ratios (p < 0.001) from 35-40 weeks. Average daily feed intake and body weight were highest (p < 0.001) in the LFE group compared to the MFE and HFE groups, while albumen height, Haugh unit and amino acid concentrations of the albumen were significantly higher in the HFE groups compared to the LFE cohort (p < 0.001). This study concludes that FE status established in early lay is a stable variable until at least 40 weeks of age, and overweight, mid-laying hens that had poor FE produced inferior egg albumen quality measurements and composition. The distinct traits of the highly efficient hens and the poor feed efficient hens may provide important information to improving productivity in egg production.

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.

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.

Short- and medium-term survival after TAVI: Clinical predictors and the role of the FRANCE-2 score.

AIM: The aim of this study was to explore the value of the FRANCE-2 score in associating with clinical outcome in the medium and short-term after TAVI and to compare its relative merits with other risk score models. METHODS: 187 consecutive patients undergoing TAVI in a single UK centre were retrospectively studied. The FRANCE-2, logistic EuroSCORE, EuroSCORE II, German AV and STS/ACC TVT risk scores were calculated retrospectively and c-statistics associating with mortality were applied. Survival outcomes were compared between different risk groups according to the FRANCE-2 scores. RESULTS: Of the 187 patients, 57.2% were male and their mean age was 80.9 ± 6.9 years. The c-index of FRANCE-2 score for predicting 30-day mortality was 0.793 (p = 0.009), for 1-year mortality 0.679 (p = 0.016) and for 2-year mortality was 0.613 (p = 0.088). The mean survival time for patients with a high FRANCE-2 score (18.6 months) was significantly less than for patients with low and moderate scores (p = 0.0004). The logistic EuroSCORE and EuroSCORE II were poorly associated with 30-day and 1-year mortality. STS/ACC TVT score was best predictive of 1-year mortality and German AV score was moderately predictive of 30-day mortality. CONCLUSIONS: The FRANCE-2 risk score is associated with differential short- and medium-term survival in patients undergoing TAVI. The presence of a high FRANCE-2 score (>5) is associated with poor survival. The FRANCE-2 scoring system could be considered as a useful additional tool by the Heart multidisciplinary team (MDT) in identifying patients who are likely to have limited survival benefit although this requires further prospective evaluation.

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

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.