Microbiome Changes in Layer Pullets Reared in Floor Pens along the Growth Period.

The gastrointestinal tract microbiome is essential for regulating nutrient absorption, gut immune function, and host growth and development. In the present study, we characterized the development of ileum and cecum microbiota in pullets throughout the rearing period, encompassing a period from the day of hatching to 18 weeks of age. The growth performance and intestinal microbiome (ileum and cecum) of pullets were analyzed at 1, 5, 11, and 18 weeks of age. The richness of the ileum and cecum bacterial communities (alpha diversity) was higher in pullets at 18 weeks of age than in those at 1 and 5 weeks of age. Microbiota from weeks 1, 5, 11, and 18 were distinctly grouped in a NMDS plot, representing beta diversity within the ileum. However, the results for cecum microbiota did not reveal evident separation among the different age groups in the weighted UniFrac. In conclusion, our findings demonstrate variations and diversification in ileum and cecum microbiota across different rearing stages in pullets. These insights have the potential to inform the development of nutritional strategies that promote gut health and contribute to the improved development of pullets.

n-3 essential fatty acid and vitamin D supplementation improve skeletal health in laying hens.

Keel bone fractures and osteoporosis are prevalent and damaging skeletal issues in the laying hen industry. There is a large interest in improving bone quality parameters to reduce or eliminate these conditions, thus improving bird welfare. Both essential fatty acids (EFA) and vitamin D can play a role in bone metabolism. The hypothesis of this study was that birds supplemented with lower n-6:n-3 EFA ratio or vitamin D would have improved bone properties compared to a control diet. A total of 3,520 Lohmann Brown-Lite pullets were used in this study. Pullets were housed on the floor from 0 to 17 wk of age and then moved to an aviary (17-52 wk of age). Starting at 12 wk of age, birds were split into diet treatments-control, flax, fish, or vitamin D diets with n-6:n-3 ratios of 6.750, 0.534, 0.534, and 6.750, respectively. Diets were formulated to be isonitrogenous and isocaloric. Basal vitamin D3 levels were formulated to be 2,760 IU/kg across all diets; for the vitamin D diet, the vitamin D3 level was increased to 5,520 IU/kg. Hens on fish and vitamin D diets had greater bone density, keel bone volume, digital bone mineral content, and keel condition compared to flax and control hens. Additionally, birds fed the vitamin D diet had the heaviest body weights compared to birds fed fish or control diets. Birds fed the flax and vitamin D diets had improved feather coverage across multiple body regions. Feeding an n-3 EFA- or vitamin D-enriched diet decreased mortality by 1.6 to 3.3% compared to the control. The fish and vitamin D diets generated mixed production performance. Compared to the other treatments, the vitamin D diet generated higher case weights but lower hen day percentage throughout the study. When compared to the other treatments, the fish diet had the lowest case weights but had a greater hen day percentage after 36 wk of age. Results indicate that a fish-based EFA and vitamin D supplementation show promise in improving skeletal health but require further investigation.

Effect of Bacillus subtilis and Oregano Oil on Performance, Gut Microbiome, and Intestinal Morphology in Pullets.

This study aimed to examine the effect of probiotics containing Bacillus subtilis and oregano essential oil on the growth performance, intestinal morphology, and cecal microbial composition in pullets aged 0-18 weeks. A total of 324 day-old Hy-Line Brown chicks were randomly assigned to three treatment groups, with six replicates per group and 18 birds per pen. The experimental treatments consisted of the following: a control group receiving a basal diet (Con), a group receiving a basal diet supplemented with 1 g/kg (3 × 108 cfu/kg) of Bacillus subtilis (BS), and a group receiving a basal diet supplemented with 0.3 g/kg of oregano essential oil (ORO). The groups supplemented with BS and ORO demonstrated significantly higher villus height/crypt depth ratios than the Con group. Microbial richness was significantly higher in groups supplemented with BS (p = 0.0317) and ORO (p = 0.00794) than in the Con group. These findings revealed a distinct separation between gut microbial communities of the Con group and those supplemented with ORO, based on unweighted and weighted UniFrac indices. Therefore, supplementation with Bacillus subtilis and oregano oil improved the composition of the microbiota, suggesting their positive effects on the gut health of pullets.

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.

Effects of full-fat high-oleic soybean meal in layer diets on nutrient digestibility and egg quality parameters of a white laying hen strain.

This study was conducted to understand the impact of including full fat high-oleic soybean meal in layer hen diets on nutrient digestibility and added nutritional value in eggs. Forty-eight layers (∼36 wk old) were randomly assigned to one of 4 isonitrogenous (18.5% crude protein) treatment diets with 12 replicate birds per treatment in a 3-wk study. Treatments were 1) solvent extracted defatted soybean meal + corn diet, 2) dry extruded defatted soybean meal + corn, 3) full-fat soybean meal + corn, 4) high-oleic full-fat soybean meal + corn diet. Apparent ileal digestibility of crude fat (CF) and crude protein (CP) were determined using celite (∼2%) as an indigestible marker. Tibia strength and egg quality parameters (egg weight, shell strength, Haugh unit, shell color, and yolk color) were recorded during the study. Fatty acid profiles, including the monounsaturated fatty acid, oleic acid (C18:1, cis), in eggs and adipogenic tissue (liver, muscle, and fat pad) were measured using gas chromatography (GC-FID). Digestibility values of CF ranged from 71 to 84% and CP varied from 67 to 72% for treatment diets, with treatment mean values being no different (P > 0.05) between treatment diets. No differences between treatment diets in tibia strength or egg quality parameters (egg weight, shell strength, and Haugh unit) were observed (P > 0.05) except for yolk color. Similarly, there were no differences in the total lipids in egg yolk (P > 0.05) between treatment diets. However, oleic acid percentage of total lipid in egg and tissue was significantly higher (P < 0.001) in hens given the high-oleic full-fat soybean meal diet than in other treatment groups. No difference was observed in oleic acid percentage of total lipid in egg between the other 3 treatment diets (P > 0.05). Overall, the results exhibited that the eggs and tissue of layer hens fed the full-fat high-oleic acid soybean meal diet were higher in oleic acid while the CF and CP digestibility remained similar to the digestibility of the other diets.

Campylobacter bilis, the second novel Campylobacter species isolated from chickens with Spotty Liver Disease, can cause the disease.

Spotty Liver Disease (SLD) is a significant disease of commercial layer hens. It can cause up to 10 % flock mortalities and reduce egg production by 25 %. Campylobacter hepaticus has been identified as the main cause of the disease, although it also appears that predisposing factors, such as some form of stress, may increase the likelihood of clinical disease occurring. Recently, a newly identified species, Campylobacter bilis, was isolated from bile samples of clinical SLD affected chickens. To investigate the pathogenic potential of C. bilis two independent isolates were used in infection trials of layer hens. Within 6 days of oral challenge birds developed typical SLD liver lesions, demonstrating that both strains induced SLD. C. bilis could be recovered from all the challenged birds that developed SLD. Thus, each of the steps in Koch's postulates have been fulfilled, confirming that C. bilis is an additional cause of SLD. A PCR method was developed which can specifically detect C. bilis from samples with complex microbiota. The identification of this newly discovered Campylobacter species as a second cause of SLD and the provision of a rapid method to detect the SLD causing bacterium will help with SLD vaccine development and epidemiology, thus assisting in the control of this important disease of poultry.

Intestinal microbiota of layer hens and its association with egg quality and safety.

The intestinal microbiota has attracted tremendous attention in the field of the poultry industry due to its critical role in the modulation of nutrient utilization, immune system, and consequently the improvement of the host health and production performance. Accumulating evidence implies intestinal microbiota of laying hens is a potential mediator to improve the prevalent issues in terms of egg quality decline in the late phase of laying production. However, the regulatory effect of intestinal microbiota on egg quality in laying hens remains elusive, which requires consideration of microbial baseline composition and succession during their long lifespans. Notable, although Firmicutes, Bacteroidetes, and Proteobacteria form the vast majority of intestinal microbiota in layer hens, dynamic intestinal microbiota succession occurs throughout all laying periods. In addition to the direct effects on egg safety, intestinal microbiota and its metabolites such as short-chain fatty acids, bile acids, and tryptophan derivatives, are suggested to indirectly modulate egg quality through the microbiota-gut-liver/brain-reproductive tract axis. These findings can extend our understanding of the crosstalk between intestinal microbiota and the host to improve egg quality and safety. This paper reviews the compositions of intestinal microbiota in different physiological stages of laying hens and their effects on egg quality and proposes that intestinal microbiota may become a potential target for modulating egg quality and safety by nutritional strategies in the future.

Technological and Sensory Quality and Microbiological Safety of RIR Chicken Breast Meat Marinated with Fermented Milk Products.

The aim of the study was to determine the effect of marinating with fermented milk products (buttermilk and sour milk) on the physical characteristics, microbiological quality, and sensory acceptability of Rhode Island Red (RIR) hen meat after the first year of laying use. The hen breast meat was marinated with fermented dairy products, buttermilk and sour milk, by the immersion method for 12 h at 4 °C. The assessed features included the quality of raw and roasted marinated and non-marinated meat in terms of physical characteristics (marinade absorption, water absorption, pH, L*, a*, b* colour, shear strength, texture profile analysis (TPA) test), microbiological parameters, and sensory characteristics. Bacteria were identified by the mass spectrometry method (MALDI-TOF MS Biotyper). Marinating meat with fermented dairy products lightened the colour, decreased the value of shear force, reduced hardness and chewiness, and limited the growth of aerobic bacteria and Pseudomonas spp. Additionally, after heat treatment, the number of identified aerobic bacteria families in the marinated in buttermilk and marinated in sour milk groups was smaller than in the non-marinated muscle group. The sensory evaluation showed a beneficial effect of marinating with buttermilk and sour milk on the tenderness, juiciness, and colour of roasted meat.

Effects of Enterococcus faecalis administration on the community structure of airborne bacteria in weanling piglet and layer hen houses.

Probiotics have been shown to improve microbial compositions in animal intestine and feces, but the effects of probiotic administration on airborne microbial composition in animal houses remain unclear. In this study, we investigated the effects of dietary Enterococcus faecalis on the bacterial community structure in the air of piglet and layer hen houses. Indoor air and feces from piglet and layer hen houses were sampled after supplementing E. faecalis in feed for 60 days, and bacterial community structures were analyzed using Illumina high-throughput sequencing technology. Results showed that Chao1, ACE, Shannon, and Simpson indices of bacterial diversity did not significantly change in feces or indoor air of piglet or layer hen after supplementation with E. faecalis (P > 0.05). However, E. faecalis administration resulted in a decrease in the relative abundance of Proteobacteria (P < 0.05). In addition, E. faecalis significantly reduced the relative abundance of opportunistic pathogens such as Acinetobacter, Escherichia, and Shigella (P < 0.05), and beneficial bacterial genus such as Lactobacillus was significantly enriched in both feces and indoor air (P < 0.05). These changes should be of benefit to livestock, farm workers, and the surrounding environment.

Amino acid requirements for laying hens: a comprehensive review.

The main aim of this review is to consolidate the relevant published data examining amino acid requirements of layer hens and to reach a new set of recommendation based on these data. There are inconsistences in lysine, sulphur-containing amino acids, threonine, tryptophan, branched-chain amino acids, and arginine recommendations in data that have surfaced since 1994. This review finds that breed, age, basal diet composition, and assessment method have contributed toward inconsistencies in amino acid recommendations. Presently, the development of reduced-protein diets for layer hens is receiving increasing attention because of the demand for sustainable production. This involves quite radical changes in diet composition with inclusions of nonbound, essential and nonessential amino acids. Increasing inclusions of nonbound amino acids into layer diets modifies protein digestive dynamics, and it may influence amino acid requirements in layer hens. This review considers present amino acid recommendations for layer hens and proposes refinements that may better serve the needs of the layer industry in the future.

Genome-wide association studies for mottled eggs in chickens using a high-density single-nucleotide polymorphism array.

Mottled eggs in layer chickens are gaining increasing attention because of the economic impact on the egg industry caused by the reduced sale value of commodity eggs. However, the genetic architecture underlying mottled eggs is not well understood. The genetic architecture underlying the mottled egg trait was investigated using genome-wide association studies (GWAS) by high-density arrays, using a total of 407 pink eggs and 799 blue eggs from an F2 resource population generated by crossing Dongxiang Blue-shelled and White Leghorn chickens. The mottled egg score in blue eggs was found to be higher than that in pink eggs. The single-nucleotide polymorphism heritability of mottled egg at laying day and storage for 7 days was 0.18 and 0.20, respectively. Bivariate GWAS provided 29 significant loci, mainly located on GGA2, GGA3, GGA8, GGA10, GGA15, GGA17, and GGA23, affecting mottled egg on laying day. Candidate genes RIMS2, SLC25A32, RIMBP2, VPS13B, and RGS3 were obtained for mottled eggshell by bivariate GWAS and gene annotation. Our findings provide new insights into the genetic architecture of mottled egg in hens, and demonstrate that a genomic selection method would be profitable for breeding out the mottled egg trait.

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

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

RESUMO: Laringotraqueíte infecciosa (LTI), causada por um alfaherpesvírus (herpesvírus Gallid-1; GaHV-1), foi observada na região das Terras Altas da Mantiqueira, Minas Gerais. De 2010 a 2018, o Serviço Veterinário Oficial (SVO) do Instituto Mineiro Agropecuário (IMA) implementou medidas para impedir a disseminação do vírus para outras regiões do estado e controlar a doença na região interditada. Devido à proximidade e consequente vínculo epidemiológico entre as granjas, a região foi considerada uma unidade epidemiológica única. Para verificar a eficiência das medidas de controle de LTI, foram realizados: (1) pesquisa soroepidemiológica, (2) questionários para avaliar medidas de biosseguridade; e (3) avaliação da influência da densidade populacional da granja na ocorrência de LTI. Em 2016, 2017 e 2018, a LTI foi investigada usando métodos epidemiológicos e clínico-patológicos, com a detecção molecular de GaHV-1. O levantamento sorológico foi realizado em 24 granjas da região interditada e em 13 granjas de outras regiões do estado. Em 2010 e 2018, foram aplicados questionários para coletar dados e determinar indicadores de medidas de biosseguridade em todas as granjas da área interditada. As diferenças foram avaliadas (p 0,05 de Wilcoxon). Os resultados indicaram sorologia positiva em toda a região, embora apenas em quatro granjas (16,6%) as galinhas apresentaram sinais clínicos, lesões macroscópicas e histológicas da LTI. A prevalência de infecção viral aumentou de 2016 (27%) para 2017 (50%) e foi maior em fazendas com alta densidade de alojamento (p=0,033). Presença da doença, vírus ou anticorpos foram detectados nas granjas fora da área interditada. Embora os indicadores de biosseguridade tenham melhorado em todas as fazendas da área interditada (p 0,05), o vírus está ativo e circulava na região. As medidas de contingência contiveram o surto, mas as práticas de biosseguridade são fundamentais para o controle de novos surtos. O controle oficial será mantido na região, incluindo a vigilância de novos casos e procedimentos de biosseguridade para mitigar o risco de transmissão do vírus para outras regiões.

Evaluation of actions of the official veterinary service to mitigate outbreaks of infectious laryngotracheitis and improve biosecurity on laying hen farms / Avaliação de ações do serviço veterinário oficial para bloqueio de foco de laringotraqueite infecciosa e melhoria da biosseguridade em granjas de postura

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

Laringotraqueíte infecciosa (LTI), causada por um alfaherpesvírus (herpesvírus Gallid-1; GaHV-1), foi observada na região das Terras Altas da Mantiqueira, Minas Gerais. De 2010 a 2018, o Serviço Veterinário Oficial (SVO) do Instituto Mineiro Agropecuário (IMA) implementou medidas para impedir a disseminação do vírus para outras regiões do estado e controlar a doença na região interditada. Devido à proximidade e consequente vínculo epidemiológico entre as granjas, a região foi considerada uma unidade epidemiológica única. Para verificar a eficiência das medidas de controle de LTI, foram realizados: (1) pesquisa soroepidemiológica, (2) questionários para avaliar medidas de biosseguridade; e (3) avaliação da influência da densidade populacional da granja na ocorrência de LTI. Em 2016, 2017 e 2018, a LTI foi investigada usando métodos epidemiológicos e clínico-patológicos, com a detecção molecular de GaHV-1. O levantamento sorológico foi realizado em 24 granjas da região interditada e em 13 granjas de outras regiões do estado. Em 2010 e 2018, foram aplicados questionários para coletar dados e determinar indicadores de medidas de biosseguridade em todas as granjas da área interditada. As diferenças foram avaliadas (p<0,05 de Wilcoxon). Os resultados indicaram sorologia positiva em toda a região, embora apenas em quatro granjas (16,6%) as galinhas apresentaram sinais clínicos, lesões macroscópicas e histológicas da LTI. A prevalência de infecção viral aumentou de 2016 (27%) para 2017 (50%) e foi maior em fazendas com alta densidade de alojamento (p=0,033). Presença da doença, vírus ou anticorpos foram detectados nas granjas fora da área interditada. Embora os indicadores de biosseguridade tenham melhorado em todas as fazendas da área interditada (p<0,05), o vírus está ativo e circulava na região. As medidas de contingência contiveram o surto, mas as práticas de biosseguridade são fundamentais para o controle de novos surtos. O controle oficial será mantido na região, incluindo a vigilância de novos casos e procedimentos de biosseguridade para mitigar o risco de transmissão do vírus para outras regiões.(AU)

Challenges in Vaccinating Layer Hens against Salmonella Typhimurium.

Salmonella Typhimurium is among the most common causes of bacterial foodborne gastrointestinal disease in humans. Food items containing raw or undercooked eggs are frequently identified during traceback investigation as the source of the bacteria. Layer hens can become persistently infected with Salmonella Typhimurium and intermittently shed the bacteria over the course of their productive lifetime. Eggs laid in a contaminated environment are at risk of potential exposure to bacteria. Thus, mitigating the bacterial load on farms aids in the protection of the food supply chain. Layer hen producers use a multifaceted approach for reducing Salmonella on farms, including the all-in-all-out management strategy, strict biosecurity, sanitization, and vaccination. The use of live attenuated Salmonella vaccines is favored because they elicit a broader host immune response than killed or inactivated vaccines that have been demonstrated to provide cross-protection against multiple serovars. Depending on the vaccine, two to three doses of Salmonella Typhimurium vaccines are generally administered to layer hens within the first few weeks. The productive life of a layer hen, however, can exceed 70 weeks and it is unclear whether current vaccination regimens are effective for that extended period. The objective of this review is to highlight layer hen specific challenges that may affect vaccine efficacy.

Improving effects of Epimedium flavonoids on the selected reproductive features in layer hens after forced molting.

In the present study, for the purpose of investigating the effects of the total flavonoids of Epimedium (TFE) in regard to preventing the development of atrophied oviducts and follicles induced by forced molting, 300-day-old Hy-Line Brown layer hens were divided into 3 study groups as follows: the control (CON) group was the normal group, without forced molting and TFE treatments; the TFE1 group was treated by adding a 1‰ TFE treatment after forced molting; and the TFE0 group was not treated by TFE after forced molting. During this study's experimental process, the egg production rates were recorded each day. In addition, the hens were randomly chosen to be weighed every 4 D and also randomly selected to be sacrificed every 7 D. Then, sample tissues of albumen-secreting part and uterus from the fallopian tube of the layer hens were collected for PCR and hematoxylin-eosin staining tests. The results showed that the body weights, number of follicles, and weights and sizes of the fallopian tube for the TFE1 and TFE0 groups were significantly reduced when compared with those of the control group on the 15th D of the experiment. Furthermore, at the end of study, it was found that the egg production rates, weights of the fallopian tube, and ovarian follicles of TFE1 had recovered to normal levels. At the same time, the serum estrogen and the expressions of the progesterone receptor and estrogen receptor mRNA in fallopian tube were higher than those observed for the TFE0 group. The results of this study provided valuable evidence that TFE could improve the development of atrophied oviducts and increase the egg laying rates, thereby making it a potential multicomponent natural drug for egg production in the future.

Salmonella on Australian cage egg farms: Observations from hatching to end of lay.

Single-aged caged layer hen flocks were monitored for Salmonella over the course of their lifetime. Chicks from both flocks were Salmonella negative at hatch and remained negative during rearing. Pullets were transported to production farms at 15 weeks of age. Pre-population dust swabs collected from both production sheds had a high percentage of Salmonella positive samples (80 and 90%). Flocks were sampled at regular intervals until 70-72 weeks of age. The proportion of Salmonella positive samples and mean load detected on eggs was low on both farms. Analysis of dust samples revealed that Salmonella persisted in dust over 8 weeks. Dust total moisture content and water activity appears to influence bacterial persistence. On egg grading equipment, only suction cups prior to egg washing were Salmonella positive (mean proportion Salmonella positive samples 0.13 ± 0.07; mean load of 18.6 ± 12.31 MPN/ml). An egg washing experiment demonstrated that while washing reduced the total Salmonella load from eggshell surfaces, no effect was observed for shell pores. These results demonstrate that despite environmental contamination on farm, Salmonella contamination of eggs is low and is further minimized by washing.

Saltbush (Atriplex nummularia and A. amnicola) as potential plants for free-range layer farms: consequences for layer performance, egg sensory qualities, and excreta moisture.

Highly adapted plant species suited to low rainfall conditions need to be considered for free-range poultry farms, particularly in marginal rainfall areas. A group of suitable plants are the saltbushes, in particular old man saltbush (Atriplex nummularia) can provide shelter, shade, and possibly forage. The first experiment examined whether hens eat old man saltbush while ranging. Free-range Hy-Line Brown layer hens were provided saltbush or "no" saltbush on the outdoor range throughout 11 wk of early production (16 to 27 wk). The quantity of saltbush eaten by the hens was determined by the n-alkane method. Hen interactions with saltbush were video recorded, and hen live weight, feed intake, egg production, and egg quality were measured. Hens ate the saltbush at 5% of their dietary dry matter intake. This level of saltbush intake had no influence on egg production. The video footage revealed hens actively pecked at the saltbush. The second experiment investigated the consequences of hens diluting their diet by eating increased levels of River saltbush (Atriplex amnicola). Air-dried, hammer-milled river saltbush was mixed and pelleted into a standard commercial layer diet at the following levels; 0 (control), 5, 10, 15, and 20%. Hy-Line Brown layer hens were fed the diets for 28 D (32 to 35 wk of age). Seventy-five hens were housed in individual cages, with 15 hen replicates per diet. The saltbush had no significant impact on egg production, hen live weight, or feed intake. Excreta moisture increased significantly (P < 0.001) with increased saltbush (5, 10, 15, and 20%) in the diet. Furthermore, high saltbush eggs (20%) had significantly stronger egg yolk color (P = 0.006) and thicker egg shells (P < 0.001) compared to the control eggs. Consumers were presented with the eggs from the second experiment and they preferred the high saltbush eggs (20%). In conclusion, saltbush would be a valuable plant on free-range poultry farms to provide shade and shelter, especially in those areas with marginal rainfall.

A Long-Term Efficacy Trial of a Live, Attenuated Salmonella Typhimurium Vaccine in Layer Hens.

Salmonella remains one of the most common causes of bacterial foodborne gastrointestinal disease in humans. Raw eggs or food items containing undercooked eggs are frequently identified as the source of Salmonella. Salmonella Typhimurium contamination of table eggs most commonly occurs when they are laid in a contaminated environment. Several control strategies, including vaccination, are widely used to mitigate the total Salmonella load. It is unclear, however, whether live attenuated Salmonella vaccines are efficacious over the life span of a layer hen. Live attenuated Salmonella vaccines have been favored due to their ability to illicit a strong humoral immune response. The lifespan of a layer hen ranges between 60 and 80 weeks and the long term efficacy of attenuated vaccine strains has not been investigated. In this study, commercial brown layer chicks were vaccinated at day old, 6 weeks of age, and again at 10 weeks of age with the Bioproperties VaxsafeTM STM1 aroA mutant vaccine. Birds were challenged at 18 weeks of age with Salmonella Typhimurium DT9 (MLVA 03 15 08 11 550). Feces and eggs were monitored for S. Typhimurium for 40 weeks post-infection. Birds produced a strong immune response following the final dose which was administered intramuscularly. The serum antibody response to S. Typhimurium DT9 infection did not differ between challenged groups. Fecal shedding and egg contamination was highly variable and did not differ significantly between vaccinated and unvaccinated birds that had been challenged with S. Typhimurium DT9. Total bacterial load in feces was quantified using qPCR. No significant difference was detected between unvaccinated and vaccinated birds after challenge.

Evaluation of Two Compressed Air Foam Systems for Culling Caged Layer Hens.

Outbreaks of avian influenza (AI) and other highly contagious poultry diseases continue to be a concern for those involved in the poultry industry. In the situation of an outbreak, emergency depopulation of the birds involved is necessary. In this project, two compressed air foam systems (CAFS) were evaluated for mass emergency depopulation of layer hens in a manure belt equipped cage system. In both experiments, a randomized block design was used with multiple commercial layer hens treated with one of three randomly selected depopulation methods: CAFS, CAFS with CO2 gas, and CO2 gas. In Experiment 1, a Rowe manufactured CAFS was used, a selection of birds were instrumented, and the time to unconsciousness, brain death, altered terminal cardiac activity and motion cessation were recorded. CAFS with and without CO2 was faster to unconsciousness, however, the other parameters were not statistically significant. In Experiment 2, a custom Hale based CAFS was used to evaluate the impact of bird age, a selection of birds were instrumented, and the time to motion cessation was recorded. The difference in time to cessation of movement between pullets and spent hens using CAFS was not statistically significant. Both CAFS depopulate caged layers, however, there was no benefit to including CO2.

Salmonella spp. contamination in commercial layer hen farms using different types of samples and detection methods.

The performance of detection methods (culture methods and polymerase chain reaction assay) and plating media used in the same type of samples were determined as well as the specificity of PCR primers to detected Salmonella spp. contamination in layer hen farms. Also, the association of farm characteristics with Salmonella presence was evaluated. Environmental samples (feces, feed, drinking water, air, boot-swabs) and eggs were taken from 40 layer hen houses. Salmonella spp. was most detected in boot-swabs taken around the houses (30% and 35% by isolation and PCR, respectively) follow by fecal samples (15.2% and 13.6% by isolation and PCR, respectively). Eggs, drinking water, and air samples were negative for Salmonella detection. Salmonella Schwarzengrund and S. Enteritidis were the most isolated serotypes. For plating media, relative specificity was 1, and the relative sensitivity was greater for EF-18 agar than XLDT agar in feed and fecal samples. However, relative sensitivity was greater in XLDT agar than EF-18 agar for boot-swab samples. Agreement was between fair to good depending on the sample, and it was good between isolation and PCR (feces and boot-swabs), without agreement for feed samples. Salmonella spp. PCR was positive for all strains, while S. Typhimurium PCR was negative. Salmonella Enteritidis PCR used was not specific. Based in the multiple logistic regression analyses, categorization by counties was significant for Salmonella spp. presence (P-value = 0.010). This study shows the importance of considering different types of samples, plating media and detection methods during a Salmonella spp. monitoring study. In addition, it is important to incorporate the sampling of floors around the layer hen houses to learn if biosecurity measures should be strengthened to minimize the entry and spread of Salmonella in the houses. Also, the performance of some PCR methods and S. Enteritidis PCR should be improved, and biosecurity measures in hen farms must be reinforced in the region of more concentrated layer hen houses to reduce the probability of Salmonella spp. presence.