Inclusion of up to 20% Black Soldier Fly larvae meal in broiler chicken diet has a minor effect on caecal microbiota.

Background: The Black Soldier Fly larvae (BSFL) are a source of nutrients and bioactive compounds in broiler diets. Some components of the BSFL may serve as a prebiotic or may impact the intestinal microbiota of the broilers by other modes of action, which in turn can affect the health and performance of broilers. Here, we investigate the impact of up to 20% BSFL in broiler diets on the diversity and composition of the broiler's microbiota. Methods: Four hundred broilers were fed five iso-nutritious experimental diets with increasing levels of BSFL meal reaching 0%, 5%, 10%, 15%, 20% in the finisher diets. Eight caecal content samples coming from each of the eight replicates per treatment were collected at two time points (day 21 and day 42) for DNA extraction and sequencing of the V3-V4 regions using Illumina MiSeq 2 × 300 bp pair-end sequencing with 341f and 805r primers. Analysis of variance and Spearman's correlation were performed, while QIIME2, DADA2, and Calypso were used for data analysis. Results: When broilers were 21 days of age, the abundance of two groups of sequence variants representing Enterococcus and unclassified Christensenellaceae was significantly lower (p-value = 0.048 and p-value = 0.025, respectively) in the 20% BSFL group compared to the 0% BSFL group. There was no relevant alteration in the microbiota diversity at that stage. On day 42, the Spearman correlation analysis demonstrated that the sequence variants representing the genus Coprococcus showed a negative relationship with the BSFL inclusion levels (p-value = 0.043). The sequence variants representing the genus Roseburia and Dehalobacterium demonstrated a positive relationship with the BSFL dietary inclusion (p-value = 0.0069 and p-value = 0.0034, respectively). There was a reduction in the dissimilarity index (ANOSIM) caused by the 20% BSFL dietary inclusion. Conclusion: The addition of up to 20% BSFL in broiler diets did not affect the overall caeca microbiota diversity or composition at day 21. On day 42, there was a reduction in the beta diversity caused by the 20% BSFL dietary inclusion. The abundance of the bacterial group Roseburia was increased by the BSFL dietary inclusion, and it may be beneficial to broiler immunity and performance.

An Initial Study on the Use of Machine Learning and Radio Frequency Identification Data for Predicting Health Outcomes in Free-Range Laying Hens.

Maintaining the health and welfare of laying hens is key to achieving peak productivity and has become significant for assuring consumer confidence in the industry. Free-range egg production systems represent diverse environments, with a range of challenges that undermine flock performance not experienced in more conventional production systems. These challenges can include increased exposure to parasites and bacterial or viral infection, along with injuries and plumage damage resulting from increased freedom of movement and interaction with flock-mates. The ability to forecast the incidence of these health challenges across the production lifecycle for individual laying hens could result in an opportunity to make significant economic savings. By delivering the opportunity to reduce mortality rates and increase egg laying rates, the implementation of flock monitoring systems can be a viable solution. This study investigates the use of Radio Frequency Identification technologies (RFID) and machine learning to identify production system usage patterns and to forecast the health status for individual hens. Analysis of the underpinning data is presented that focuses on identifying correlations and structure that are significant for explaining the performance of predictive models that are trained on these challenging, highly unbalanced, datasets. A machine learning workflow was developed that incorporates data resampling to overcome the dataset imbalance and the identification/refinement of important data features. The results demonstrate promising performance, with an average 28% of Spotty Liver Disease, 33% round worm, and 33% of tape worm infections correctly predicted at the end of production. The analysis showed that monitoring hens during the early stages of egg production shows similar performance to models trained with data obtained at later periods of egg production. Future work could improve on these initial predictions by incorporating additional data streams to create a more complete view of flock health.

Prevalence and magnitude of gastrointestinal helminth infections in cage-free laying chickens in Australia.

Helminth infections have been re-emerging with the growing popularity of free-range and floor-based chicken production systems. The aim of this study was to determine the prevalence and worm burdens of intestinal helminth infection in cage-free laying chickens in Australia. In an online survey about worm prevalence, a high proportion of respondents reported the detection of Ascaridia galli (77%), followed by tapeworms (69%) and caecal worms (Heterakis gallinarum) (62%), whereas fewer respondents (23%) reported the presence of hair worms (Capillaria spp.) in their flocks. Total worm recovery from 407 laying hens on four farms found that 92.1% of hens harboured one or more helminth parasite with a prevalence of 73 to 100% across farms. Mixed infections were common with 79% of hens harbouring two or more helminth species. The prevalence of nematode species H. gallinarum, A. galli and Capillaria spp. was 87, 82 and 35% respectively. Five cestode species were found with a low individual chicken prevalence (Raillietina tetragona 4.7%, Raillietina echinobothrida 3.2%, Raillietina cesticillus 5.2%, Choanotaenia infundibulum 4.4%, and Hymenolepis cantaniana 4.4%). The hens harboured an average of 71 worms with H. gallinarum having the highest mean burden (45.5 worms/hen) followed by A. galli (22.0 worms/hen), Capillaria spp. (2.7 worms/hen) and cestodes (0.8 worms/hen). The sex ratio (female:male worms) was 1.38:1 for A. galli, and 1.77:1 for H. gallinarum. There was a strong positive correlation between A. galli female worm count and excreta egg count (EECs) (rs = 0.94, P < 0.0001) and also between total nematode worm count and EEC (rs = 0.82, P < 0.0001) in individual hens. When investigating intestinal excreta (n = 10) and caecal excreta (n = 10) of 16 chicken flocks the prevalence of infection with ascarid worms in intestinal and caecal excreta was 71 and 78% respectively and 27% prevalence of Capillaria spp. in intestinal excreta with mean EECs of 407, 404, and 18 eggs/g of excreta (EPG), respectively. These results suggest that most chickens kept in free-range or floor production systems are infected with one or more helminth parasite species. Heavy worm infections would likely affect the production performance and welfare of birds with adverse economic impact. Strategic or tactical anthelmintic treatment with effective anthelmintic could reduce this impact.

Ascaridia galli eggs obtained from fresh excreta, worm uteri or worms cultured in artificial media differ in embryonation capacity and infectivity.

Ascaridia galli infection models use eggs isolated from chicken excreta, worm uteri and worms cultured in artificial media. The aim of this study was to compare the infectivity of A. galli eggs isolated from these sources under two infection regimens. A 3 × 2 factorial arrangement was employed to test the infectivity of A. galli eggs from the three sources and two modes of infection (single or trickle infection). One hundred and fifty-six Isa-Brown one day-old cockerels randomly assigned to the six treatment groups (n = 26) were orally infected with embryonated A. galli eggs obtained from the three A. galli egg sources (worm uteri, excreta or eggs shed in vitro) administered either as single dose of 300 eggs at one day-old or trickle infected with 3 doses of 100 eggs over the first week of life. Twenty-two negative control birds remained uninfected. Eggs obtained from cultured worms or excreta exhibited a higher embryonation capacity (P = 0.003) than eggs obtained from worm uteri. There were higher worm establishment (infectivity) rates from embryonated eggs originating from cultured worms and worm uteri compared with eggs obtained from fresh excreta (P < 0.0001). Trickle infection resulted in a significantly higher total worm burden (P = 0.002), establishment rate (P = 0.002) and excreta egg counts (EEC, P = 0.025) than single infection. Worm length was greater in birds infected with embryonated eggs from excreta than from uteri or cultured worms (P < 0.0001). However, mode of infection did not affect worm length (P = 0.719) and weight (P = 0.945). A strong significant positive linear correlation was observed between EECs and female worm counts at 12 weeks of post infection sampling (r = 0.75; P < 0.0001). Body weight of birds was negatively correlated with both worm burden (r = - 0.21; P < 0.01) and EEC (r = - 0.20; P < 0.05) at 12 weeks post infection. In conclusion, our results show that eggs shed by cultured worms or isolated from worm uteri had greater infective capacity than eggs harvested from excreta and that trickle rather than bolus infection resulted in higher worm establishment. These factors should be taken into account when considering artificial infection protocols for A. galli.

Method optimisation for prolonged laboratory storage of Ascaridia galli eggs.

Eggs in the infective stage of the chicken nematode Ascaridia galli are often required for in vivo and in vitro studies on this parasite. The reliability of any artificial A. galli infection depends on the viability and embryonation capacity of A. galli eggs. The aim of this study was to determine ideal storage conditions for maximising the viability of A. galli eggs and maintaining viability for the longest period. A 2 × 2 × 3 × 5 factorial experimental design was employed to investigate the effects of storage temperature (4°C or 26°C), storage condition (aerobic or anaerobic), storage medium (water, 0.1 N H2SO4 or 2% formalin) and storage period (4, 8, 12, 16 and 20 weeks). The viability of eggs was assessed after eggs in all treatment groups were held aerobically at 26°C for 2 weeks after the storage period to test embryonation capacity. Based on morphological characteristics, they were categorised as undeveloped, developing, vermiform, embryonated or dead. The maintenance of viability during storage at 4°C was optimal under anaerobic conditions while at 26°C it was optimal under aerobic conditions. Anaerobic conditions at 26°C led to a rapid loss of viability while aerobic conditions at 4°C had a less severe negative effect on maintenance of viability. Egg storage in 0.1 N H2SO4 resulted in a significantly higher viability overall (54.7%) than storage in 2% formalin (49.2%) or water (37.3%) (P < 0.0001). Untreated water was the least favourable storage medium when eggs were stored at 26°C while it was a medium of intermediate quality at 4°C. The viability of A. galli eggs decreased significantly with storage time (P < 0.0001) depending on the other factors. The lowest rate of decline was seen with storage of eggs under anaerobic conditions at 4°C or aerobic conditions at 26°C in 0.1 N H2SO4. Eggs in these treatments retained up to 72% of overall viability at 20 weeks with a decline rate of approximately 2% per week with no significant difference between the two. Therefore, this study has clearly revealed opposing aerobic conditions required for prolonged storage of A. galli eggs in the pre-embryonated state at 4°C. It has also identified that 0.1 N H2SO4 provides the best preservation against degradation during storage, particularly at 26°C under aerobic conditions. Achieving strictly anaerobic conditions can be difficult to achieve so storage aerobically at 26°C may be preferred for simplicity.

Worm control practices on free-range egg farms in Australia and anthelmintic efficacy against nematodes in naturally infected layer chickens.

This study investigated worm control practices by free-range egg farmers and the efficacy of the commercial anthelmintics levamisole (LEV), piperazine (PIP), flubendazole (FLBZ) and fenbendazole (FBZ) against gastrointestinal nematodes on two free-range layer farms in Australia. An online survey comprising 36 questions was designed and implemented using SurveyMonkey. The survey contained questions about participant demographics, farm and flock characteristics, perceived intestinal worm importance, infection monitoring, deworming and other worm control practices. A link for the survey was emailed to free range egg producers from their industry body in December 2019. The anthelmintic efficacy trial was conducted in a total of 229 layers naturally infected with Ascaridia galli, Hetarakis gallinarum, Capillaria spp. and/or tape worms. Chickens received a single oral dose of LEV (28 mg/kg), PIP (100 mg/kg), FBZ (10 mg/kg) or LEV-PIP co-administered at their full individual doses, and FLBZ (Flubenol®), 30 ppm or 60 ppm) in the feed over 7 days. Anthelmintic efficacies were estimated by both worm count reduction (WCR %) and excreta egg count reduction (EECR %) tests 10 days after start of treatment. The survey with a response rate of 16/203, revealed that worm infection was of moderate concern to the producers and the majority (68%) felt that the current anthelmintics work effectively. The level of understanding of worms, monitoring and control practices did not reveal any major deficiencies of concern. The most commonly used anthelmintic was LEV (73%) followed by PIP (45%). Based on a standard cut-off value (≥90%), LEV, LEV-PIP, and FBZ attained the desired efficacy but PIP exhibited reduced efficacy against immature A. galli (61-85%), all stages of H. gallinarum (42-77%) and Capillaria spp. (25-44%). FLBZ was highly effective against all stages of roundworms and tapeworm infections. Even though there was some association between the efficacies estimated by WCR % and EECR % the latter was poorly associated in the natural infection model and hence does not provide a reasonable alternative for assessing anthelmintic efficacy when immature stages of the lifecycle are included. These results show no evidence of loss of susceptibility to the tested anthelmintics on these farms supporting the perception of producers that participated in the survey that current treatments work effectively. The reduced efficacy of PIP against some species and immature stages is related to its spectrum of activity rather than providing evidence of emerging resistance.

Evaluation of in vitro methods of anthelmintic efficacy testing against Ascaridia galli.

To investigate methods for in vitro assessment of anthelmintic efficacy against the chicken nematode Ascaridia galli this study firstly evaluated sample preparation methods including recovery of eggs from excreta using different flotation fluids and induced larval hatching by the deshelling-centrifugation method and the glass-bead method with or without bile. It then evaluated two in vitro assays, the in-ovo larval development assay (LDA) and larval migration inhibition assay (LMIA), for anthelmintic efficacy testing against A. galli using fresh eggs and artificially hatched larvae, respectively. Four anthelmintics, thiabendazole (TBZ), fenbendazole (FBZ), levamisole (LEV) and piperazine (PIP) were employed using an A. galli isolate of known susceptibility. The results suggested that the LDA and LMIA could successfully be used to generate concentration response curves for the tested drugs. The LDA provided EC50 values for inhibition of egg embryonation of 0.084 and 0.071 µg/ml for TBZ and FBZ, respectively. In the LMIA, the values of effective concentration (EC50) of TBZ, FBZ, LEV and PIP were 105.9, 6.32, 349.9 and 6.78 × 107 nM, respectively. For such in vitro studies, a saturated sugar solution showed high egg recovery efficiency (67.8%) and yielded eggs of the highest morphological quality (98.1%) and subsequent developmental ability (93.3%). The larval hatching assays evaluated did not differ in hatching efficiency but the deshelling-centrifugation method yielded larvae that had slightly better survival rates. For final standardization of these tests and establishment of EC50 reference values, tests using isolates of A. galli of defined resistance status need to be performed.

Ascaridia galli challenge model for worm propagation in young chickens with or without immunosuppression.

With the continued growth of free-range egg production, the importance of the chicken roundworm Ascaridia galli is increasing. Investigations into this parasite would be facilitated by the availability of characterised strains and clear guidelines on optimal methods of multiplication and maintenance. Currently, there is lack of well-defined in vivo models for maintaining A. galli and the potential of using host immunosuppression to boost parasite development and worm egg output has not been investigated. To determine the most efficient way of propagating A. galli in young chickens an experiment with a 2 × 3 × 4 × 2 factorial design involving age of chicken at infection (day-old or 14 days old), immunosuppression (dexamethasone (DEX), cyclophosphamide (CY) or sham), infective egg dose (0, 100, 300 or 900 embryonated eggs/bird) and time of worm recovery after infection (8 or 10 weeks post-infection) was conducted. The experiment used a total of 384 layer cockerel chicks. Infection was delivered orally in 3 split doses over one week and immunosuppressants were administered by intramuscular injection concurrently with the infections. Body weight, excreta egg counts, intestinal worm count and worm establishment rate were assessed. The only sign of ascaridiosis noted was mild diarrhoea at the time of slaughter in some birds with a significant- positive association with worm count. Infection caused a significant dose dependent reduction in body weight in non-immunosuppressed birds but this effect was ameliorated by immunosuppression. Age at infection had no significant effect on the studied variables although both worm and egg counts were numerically higher in the day-old infected groups. Egg dose significantly influenced the prevalence of infection, worm establishment rate, worm egg production and mean worm count. The 300 and 900 egg doses resulted in significantly higher worm count and egg production than the 100 egg dose. A significant negative correlation was observed between egg dose and worm establishment rate indicating an inverse relationship. Immunosuppression with DEX, but not CY resulted in significantly higher mean worm burden than in control chickens with excreta egg counts also considerably higher in DEX treated birds. Our results suggest that trickle infection at day-old with infective doses of 300 eggs coupled with immunosuppression with DEX would provide the most efficient way to propagate A. galli worms in vivo, as using older birds or a higher egg dose did not provide any advantage.

Anthelmintic efficacy evaluation against different developmental stages of Ascaridia galli following individual or group administration in artificially trickle-infected chickens.

The efficacy of commercially available anthelmintics against mature and immature stages (including ovicidal effects) of two Australian field isolates of Ascaridia galli was evaluated in two separate experiments. The anthelmintics tested were levamisole (LEV), piperazine (PIP) and flubendazole (FBZ) plus LEV-PIP. A total of 192 artificially trickle-infected young cockerels (96 birds per isolate) were randomized into sixteen experimental groups of 12 cockerels each (7 treatments and 1 untreated control per isolate). Chickens received label-recommended doses of LEV (28 mg/kg), PIP (100 mg/kg) or LEV-PIP co-administered at their full individual doses as a single oral dose or in group drinking water at recommended concentrations of 0.8 mg/ml or 2.5 mg/ml over eight hours for 1 and 2 days respectively and FLBZ (30 ppm) in the feed over 7 days. Anthelmintic efficacies were assessed by worm count reduction (WCR%) and excreta egg count reduction (EECR%) estimated by two methods. Ten days post treatment, all untreated control birds harboured mixed worm population of 10.1 and 12.3/bird for each isolate respectively which was significantly higher (P < 0.0001) than counts in all treatment groups. Luminal or histotrophic larvae comprised 50-57 % of the total worm count. For LEV, PIP and LEV-PIP, individual oral administration provided a somewhat higher efficacy than group medication in drinking water. EECR% values were inconsistent with WCR% and found to be only an indicator of efficacy against adult worms. All developmental stages of the two A. galli isolates were highly susceptible to FLBZ (100 %) followed by LEV-PIP (92.4-100 %) and LEV (87.7-100 %). PIP exhibited good efficacy against adult worms (92-97 %) but reduced efficacy against luminal (79-84 %) and histotrophic (61-72 %) larvae. Embryonation capacity of eggs recovered from worms expelled after treatment with LEV (47-54 %), PIP (44-54 %) or LEV-PIP (45-48 %) did not differ from those from untreated birds (50-51 %) whereas eggs from FLBZ treated worms had a significantly lower (P < 0.05) capacity to embryonate (≤ 2 %). Put together, our results demonstrate no evidence of resistance of the test A. galli isolates to the tested anthelmintics but a significant advantage of FLBZ, followed by LEV-PIP and LEV over PIP in the control of A. galli, specifically with regard to immature stages. A. galli worms expelled after treatment with LEV, PIP or their combination, but not FLBZ contain viable eggs. This has epidemiological implications and may also provide an option for isolating eggs from mature worms for A. galli propagation experiments without having to sacrifice birds.

Comparison of the Modified McMaster and Mini-FLOTAC methods for the enumeration of nematode eggs in egg spiked and naturally infected chicken excreta.

Excreta egg counting techniques are used for indirectly estimating the magnitude of gastrointestinal nematode infection in live animals. The aim of this study was to optimise laboratory and field sampling methods for routine monitoring of nematode infections in chickens by evaluating the sensitivity, accuracy, and precision of the Modified McMaster (MM) and Mini-FLOTAC (MF) methods using laying chicken excreta samples spiked with estimated true numbers of eggs (Experiment 1 = 5-1500 EPG (eggs/g); Experiment 2 = 5-500 EPG) without and with operator effects, respectively or using individual fresh excreta (n = 230) and fresh floor excreta (n = 42) from naturally infected free-range layer farms. The Coefficient of Variation (CV) was assessed within and between operators and the time spent on sample preparation and counting was also evaluated. MF was more sensitive than MM at ≤ 50 EPG level but not above this while MM had a significantly higher egg recovery rate than MF for ≥ 50 EPG levels (MM = 89.7 %, MF = 68.2 %; P < 0.0001). Operator factors did not have a significant effect (P = 0.358-0.998) on egg counts across methods and EPG levels. The CV between replicates of the MM and MF methods for ≥ 50 EPG was 43.4 and 36.5 %, respectively. The inter-observer CV of the MM and MF methods for ≥ 50 EPG levels was 63.8 and 44.3 % respectively. When the naturally infected free-range layers which were individual caged for excreta sampling, the proportion of samples positive for MM and MF were 91.7 and 96.5 %, respectively (P = 0.023). MM resulted in significantly (P = 0.029) higher excreta egg counts (604) than MF (460) with the difference between methods greatest at higher EPG levels. Fresh floor excreta (pooled or individual) and individual caged chicken excreta did not have significant effect on egg counts (P = 0.274). The total time taken for sample preparation and egg counting was significantly lower using the MM method (4.3-5.7 min) than the MF method (16.9-23.8 min) (P < 0.0001). In conclusion, MM was more accurate than MF, particularly at higher EPG levels, but slightly less precise and sensitive, particularly at low EPG levels, while taking less than 25 % of the laboratory time per sample. Our observations indicate that the MM method is more appropriate for rapid diagnosis of chicken nematodes in the field. Pooled fresh floor excreta samples would be sufficient to indicate infection level in free range farms.

The Response of Layer Hen Productivity and Egg Quality to an Additional Limestone Source When Offered Diets Differing in Calcium Concentrations and the Inclusion of Phytase.

Laying hens require substantial quantities of calcium (Ca) to maintain egg production. However, maintaining recommended dietary Ca through inclusion of limestone may impede nutrient digestibility, including that of other minerals. It was hypothesized that providing a separate source of dietary Ca in the form of limestone grit would preserve Ca intake of hens offered diets containing suboptimal Ca concentrations. Furthermore, the impact of dietary phytase at a "superdosing" inclusion rate on the voluntary consumption of limestone grit was evaluated. One hundred and forty-four laying hens (19 weeks of age) were assigned to one of six dietary treatments in a 3 × 2 factorial arrangement comprising three dietary Ca concentrations (40, 30, and 20 g/kg) and ±dietary phytase (3500 FYT/kg diet) on an ad libitum basis for six weeks. Limestone grit (3.4 ± 1.0 mm) was provided to all hens ad libitum. Hens offered diets containing phytase consumed significantly less limestone grit p = 0.024). Egg weight, rate of lay, and egg mass were unaffected by dietary treatment (p > 0.05). Egg shell weight % (p < 0.001), shell thickness (p < 0.001), and shell breaking strength (p < 0.01) decreased in line with dietary Ca levels. In summary, dietary superdosing with phytase reduced the consumption of a separate limestone source in individually housed, early lay ISA Brown hens. Egg shell quality variables but not egg production worsened in line with lower dietary Ca levels.

Black soldier fly larvae in broiler diets improve broiler performance and modulate the immune system.

Non-conventional feed ingredients are receiving more interest in their ability to increase farming efficiency, sustainability and animal performance. The objective of this study was to determine the optimal rate of inclusion level of the full-fat black soldier fly larvae (BSFL) in broiler diets and to evaluate their impact on performance, nutrient digestibility, and the immune system (blood cells and intraepithelial lymphocytes). A total of 400 male day-old Ross 308 broilers were randomly assigned to 5 treatment groups with 8 replicates each. Five inclusion levels of full-fat BSFL were investigated across starter (0, 2.5%, 5%, 7.5% and 10%), grower and finisher diets (0, 5%, 10%, 15% and 20%). All diets were formulated based on digestible amino acid values according to the Aviagen (2016) recommendations. A polynomial regression at different degrees was performed to analyse broiler performance parameters (body weight, body weight gain, feed intake, and feed conversion ratio), nutrient digestibility, and blood cell count. Intraepithelial lymphocyte population data was analysed performing univariate linear regression. During the entire experimental period (from 2 to 42 d), BSFL inclusion levels decreased the feed conversion ratio by 10% in broilers that received 20% BSFL in their diets (P < 0.05). Lymphocytes and white blood cell count decreased linearly by 47.7% and 35.9%, respectively, with up to 20% BSFL inclusion (P < 0.001). A 4-fold decrease in CD3+ T lymphocytes and a 9.7-fold decrease of CD3+CD8+ intestinal cytotoxic T lymphocytes occurred in broilers fed 20% BSFL compared to the control group. These findings suggest that the inclusion of BSFL can improve broiler performance and potentially reduce immune response energy expenditure in birds fed 20% BSFL for 42 d.

Comparative therapeutic efficacies of oral and in-water administered levamisole, piperazine and fenbendazole against experimental Ascaridia galli infection in chickens.

Evidence on the current efficacy status of anthelmintics used in the Australian poultry sector is lacking. A controlled trial was conducted to evaluate the efficacy of three commonly used anthelmintics, namely levamisole (LEV), piperazine (PIP) and fenbendazole (FBZ) plus levamisole-piperazine combination (LEV-PIP) against a field strain of A. galli recovered following flock treatment with LEV. A total of 108 A. galli infected cockerels were randomized into nine experimental groups of 12 cockerels each (eight treatments and one untreated control) with each treatment administered by two routes (oral drench or in drinking water). Chickens received label-recommended doses of LEV (28 mg/kg) and PIP (100 mg/kg) while LEV-PIP involved both compounds co-administered at their full individual dose rates. FBZ was tested at two dose rates; 10 mg/kg as a single oral drench or 5 mg/kg in drinking water over 5 days. Anthelmintic efficacies were assessed by worm count reduction (WCR%) and excreta egg count reduction (EECR%) estimated by two methods. Ten days post treatment, the untreated control birds harboured significantly higher worm counts (P < 0.0001) than those in all treatment groups irrespective of the mode drug of application. Oral drenching caused a greater reduction in worm and egg counts (P < 0.05) than medication in drinking water. Based on geometric worm counts the percentage efficacies for the oral drench were 99.1, 96.3, 97.2 and 100 % respectively for LEV, PIP, FBZ and LEV-PIP, and for administration in water 96.4, 93.7, 88.7 and 97.7 % respectively. Efficacies based on EECR% were consistent with WCR% with strong positive linear association between efficacy values. In conclusion, our results demonstrate no evidence of loss of susceptiblity of the test A. galli isolate to both LEV and PIP contrary to our hypothesis. Additional efficacy studies are needed using A. galli isolates sourced from different poultry flocks across Australia.

Global and regional prevalence of helminth infection in chickens over time: a systematic review and meta-analysis.

Gastrointestinal helminth parasites are a concern for the poultry industry worldwide as they can affect the health, welfare, and production performance. A systematic review of the prevalence over time in different countries may improve our understanding of gastrointestinal helminthiasis in chickens and subsequently lead to improved poultry health. The aim of this systematic review and meta-analysis was to provide an overview of the published information regarding the epidemiology and the diagnostic approaches of chicken helminth infection. Six databases were searched for studies, and a total of 2,985 articles published between 1942 and 2019 were identified and subsequently screened for eligibility using title or abstract and full text assessment, resulting in 191 publications to be used in the study. Postmortem diagnostics (73.8%) and the flotation technique (28.8%) were commonly used to detect helminth infections with a pooled prevalence of 79.4% ranging from 4 to 100%. More than 30 helminth species in chicken populations were identified including Ascaridia galli (35.9%), Heterakis gallinarum (28.5%), Capillaria spp. (5.90%), and Raillietina spp. (19.0%) being the most prevalent. The reported prevalence of helminth infection decreased over time in developing countries while it increased in the developed world. Chicken kept in backyard and free-range systems had a markedly higher pooled prevalence of helminth infection (82.6 and 84.8%, respectively) than those housed in cage production systems (63.6%). This may indicate the need for more rigorous control and prevention measures in free-range and backyard production systems using regular deworming coupled with access to early and accurate diagnosis allowing for early intervention.

Technological Quality, Amino Acid and Fatty Acid Profile of Broiler Meat Enhanced by Dietary Inclusion of Black Soldier Fly Larvae.

We evaluated the effects of full-fat black soldier fly larvae (BSFL) on broiler carcass composition, cut yield, and breast meat quality. Broilers were fed for 42 days with up to 20% dietary inclusion of BSFL (0, 5, 10, 15, and 20%). On day 42, 120 broilers were slaughtered, and images were taken using computed tomography. Breasts, drumsticks, and thighs were collected for cut yield determination. The pH, color, lipid oxidation, cooking loss, shear force, amino acid profile, and fatty acid profile of the breast meat were assessed. There was no dietary effect on carcass composition or meat quality parameters except for fatty and amino acids compositions. When 20% BSFL was included in the diet, individual fatty and amino acids, such as lauric and myristic acids, aspartic acid, glutamine, and lysine, increased by 22.0-, 5.50-, 1.08-, 1.06-, and 1.06-fold, respectively (p < 0.05). Although total polyunsaturated fatty acids decreased, eicosapentaenoic fatty acids (EPA) increased by 78% in the 20% BSFL inclusion group. In conclusion, up to 20%, dietary full-fat BSFL did not affect key meat characteristics but positively increased the levels of the health-claimable omega-3 fatty acid EPA.

Managing Free-Range Laying Hens-Part B: Early Range Users Have More Pathology Findings at the End of Lay but Have a Significantly Higher Chance of Survival-An Indicative Study.

While free-range laying hens frequently experience health and welfare challenges, the contribution of range use towards these risks are largely unknown. The aim of this pilot study was to investigate the survival, health and welfare of commercial free-range laying hens and explore the association with early range use. Range use of 9375 Lohmann Brown hens housed within five flocks was assessed during 18-21 weeks of age and individual hens were classified as "rangers" (frequent range users), "roamers" (intermittent range users), and "stayers" (rare/no range users) were then subject to necropsy at 74 weeks of age. Rangers and roamers were three times and 2.4 times more likely to survive than stayers, respectively (p = 0.001). Overall, rangers had significantly better feather cover and more lesions associated with spotty liver diseases compared to roamers and stayers (p = 0.001). Similarly, rangers and roamers had a higher prevalence of A. galli infection and less frequent signs of fatty liver syndrome compared to stayers. Rangers had a higher proportion of hens with full ovary follicle production compared to stayers and roamers (p = 0.035). This information is highly relevant to consider the targeted support of different flock subpopulations to improve hen health and welfare, directly affecting farm profitability. Further research on other farms is warranted to investigate the transferability of the observed results.

Frequent Visits to an Outdoor Range and Lower Areas of an Aviary System Is Related to Curiosity in Commercial Free-Range Laying Hens.

Individual hen preferences to spend time at particular locations within a free-range aviary system and relationships with temperament is relatively unknown. Hens (n = 769) from three commercial flocks were monitored with Radio Frequency Identification technology to determine time spent on the range, upper and lower aviary tiers, and nest boxes. Prior depopulation, novel arena (NA) and novel object (NO) tests assessed exploration and fearfulness. During early life; more time on the lower tier was associated with more lines crossed in the NA test (p < 0.05). No other evidence suggested preference during early life was related to fear or curiosity. More time on the range and lower tier were associated with heavier pre-ranging body weight and gain (p = 0.0001). Over the hens' whole life; time spent on range and lower tier was associated with approaching the NO (p < 0.01). More time spent on the upper tier was associated with less time near the NO and fewer lines crossed in NA (p < 0.01). The relationships during early and whole life use of space and some potential indicators of fearfulness were inconsistent and therefore, no strong, valid, and reliable indicators of hen fearfulness such as freezing were identified.

Body weight and range usage affect net energy utilisation in commercial free-range laying hens when evaluated in net energy chambers.

Within a given free-range flock, some hens prefer to spend the majority of their time in the shed (stayers), while others frequently access the range (rangers). Laying performance has been associated not only with the development of these sub-populations but also with different body weights (BW). The purpose of this study was to determine if range usage, BW or a combination of both is associated with energy metabolism and as such contribute to improved hen performance. Forty-eight Lohmann Brown hens at 74 wk of age were selected from a commercial free-range farm based on their BW and range usage over a 56-week period. Using a 2 × 2 factorial arrangement, hens were either classified as heavy (mean ± SEM; 2.01 ± 0.02 kg, n = 24) or light (1.68 ± 0.01 kg, n = 24), and also classified as rangers (accessed the range for 84.1% of available days, 242 ± 3.75 d; n = 24) or stayers (accessed the range for 7.17% of available days; 23.4 ± 6.08 d, n = 24). Stayers had significantly higher metabolizable energy (ME) intake per metabolic BW per d (0.852 vs. 0.798 MJ/kg BW0.75 per d; P = 0.025), higher heat production (0.637 vs. 0.607 MJ/kg BW0.75 per d; P = 0.005), higher heat increment (0.267 vs. 0.237 MJ/kg BW0.75 per d; P = 0.005) and retained more nitrogen (1.59 vs. 1.46 g/hen per d; P = 0.023) compared to the rangers. Light hens had significantly higher metabolic energy intake per metabolic BW (0.854 vs. 0.796 MJ/kg BW0.75 per d; P = 0.018), net energy (NE) intake (0.595 vs. 0.551 MJ/kg BW0.75 per d; P = 0.032), and retained energy (0.225 vs. 0.181 MJ/kg BW0.75 per d; P = 0.032), as well as lower heat production (0.936 vs. 1.003 MJ/hen per d; P = 0.002) compared to heavier hens. An interaction was observed across levels of analysis i.e. between light stayers and light rangers. The light rangers had significantly higher NE intake compared to the light stayers (9.77 vs. 9.27 MJ/kg BW0.75 per d; P = 0.024). In conclusion, light hens were more energy efficient compared to heavy hens. Moreover, light rangers had a more efficient feed utilisation compared to the light stayers.

Managing Free-Range Laying Hens-Part A: Frequent and Non-Frequent Range Users Differ in Laying Performance but Not Egg Quality.

Little is known about the impact of ranging on laying performance and egg quality of free-range hens. The aim of this study was to characterise egg production of commercial free-range laying hen sub-populations of low-, moderate- and high-range use at an early age. A total of five flocks with 40,000 hens/flock were investigated where 1875 hens/flock were randomly selected at 16 weeks of age, monitored for their range use and subsequently grouped into "stayers" (the 20% of hens that spent the least time on the range), "roamers" (the 20% of the hens that used the range more than stayers but less than rangers) and "rangers" (the 60% of the hens that spent the most time on the range). Eggs from the individual groups were collected in 10-weekly intervals until hens were 72 weeks of age, commercially graded and tested for several quality parameters. Significant differences were noted for hen-day production. For example, at 22 weeks of age, rangers enjoyed a laying rate of 88.0% ± 1.1%, while stayers performed at 78.2% ± 1.9% but at 72 weeks of age egg production of rangers was 85.1% ± 0.9% and of stayers was 95.5% ± 0.9% (p < 0.05). Range use was of minor importance to the egg quality.

Effect of pecking stones and age on feather cover, hen mortality, and performance in free-range laying hens.

Severe feather pecking is frequently associated with impaired egg production, poor hen welfare, and increased mortality. The aim of this study was to investigate the potential of pecking stones to ameliorate the incidence of feather pecking. A total of 18 flocks of Hy-Line Brown laying hens were randomly assigned to control (n = 9 no pecking stone access) or treatment (n = 9 pecking stone access) flocks and housed in commercial fixed sheds (farm A, n = 10) or commercial mobile sheds (farm B, n = 8) differing in various housing conditions. Beak length, feather score, egg production, and mortality were evaluated at 16 wk of age and every 10 wk until at least 46 wk of age. On farm A, hens with access to pecking stones had significantly lower mortality than hens without pecking stone access (P = 0.001). A significant interaction between pecking stone and age was observed on feather scores of wings of hens housed in farm A. Hens of the pecking stone group in farm A had higher wing feather score (indicating better feather condition) at the age of 56 and 66 wk than hens with no access to pecking stones (pecking stone × age, P = 0.002). The age of the hens was significantly associated with lower overall feather scores (poorer feather condition), reduced egg production, and higher mortality (P < 0.05). Although pecking stones reduced some feather loss and mortality, this effect was only present on one farm and therefore may be related to farm management. Especially the impact of pecking stones on mortality was inconclusive as the cumulative mortality in farm B was nearly twice as high compared with that in control flocks. Further investigations are warranted including the effects of pecking stone provision at an early age such as during rearing (0-17 wk of age).