Bromelain can reduce the negative effects of a subclinical necrotic enteritis in broiler chickens.

This study was conducted to examine the efficacy of a bromelain-based supplementation coded ANR-pf on growth performance and intestinal lesion of broiler chickens under necrotic enteritis (NE) challenge. A total of 540 Ross 308 day-old male chicks were randomly allocated into 6 treatments of 6 replicates. The bromelain formulation was delivered to chickens through gavaging or in drinking water method twice, on d 8 and 13. Nonchallenged groups included 1) without or 2) with the specific bromelain formulation gavaged at 0.8 mL/kg. NE-challenged groups included 3) without the specific bromelain formulation; 4) gavaged with 0.4 mL/kg; 5) gavaged with 0.8 mL/kg and 6) supplemented with 0.8 mL/kg via drinking water. Birds were challenged with Eimeria spp. on d 9 and Clostridium perfringens (NE-18 strain) on d 14 and 15. On d 14 and 19, fresh faecal contents were collected for the determination of oocyst counts. Intestinal lesion scores were determined on d16. Performance and mortality were recorded throughout the entire experiment. Among challenged groups, birds received additive via drinking water had higher weight gain (WG) compared to the remaining groups (P < 0.001) in the grower phase and had lower FCR compared to 0.4 mL/kg inoculated group in the grower and finisher phases (P < 0.001). Bromelain supplementation via drinking water improved the WG of challenged birds, similar to that of the nonchallenged birds (P < 0.001), and lowered FCR compared to other challenged groups (P < 0.001). Nonchallenged birds and birds that received bromelain formulation in drinking water did not have lesions throughout the small intestine whereas challenged birds, either un-supplemented or supplemented with bromelain via inoculation route recorded similar lesion score levels in the jejunum. At d 19, birds received bromelain in drinking water had lower fecal oocyst numbers compared to challenged birds without additive (P < 0.001). In conclusion, bromelain administration via drinking water could ameliorate the negative impacts of NE-infection in broilers by improving performance, lowering the oocyst numbers and lesion scores.

The effect of sex and dietary crude protein level on nutrient transporter gene expression and cecal microbiota populations in broiler chickens.

It is well known that male and female broilers differ in their growth performance and that many physiological factors contribute to this difference. The aim of this experiment is to investigate if there are differences between male and female broilers in cecal microbiota and nutrient transporter gene expression and if these differences play a role in the growth performance of broilers. The possible effect of protein level and its interaction with sex on microbiota and expression of the nutrient transporters were also investigated. Samples were collected from male and female birds fed either standard crude protein (SCP) or reduced crude protein diets (RCP) at the age of d 35. The experiment was designed as a 2 × 2 factorial arrangement of treatments consisting of 448 Cobb 500 broilers assigned to 32-floor pens with 4 treatments, 8 replicates, and 14 birds per pen for performance measurements. The factors were sex (male or female) and dietary crude protein (CP) level (standard or reduced). Body weight gain (BWG), feed intake and feed conversion ratio were recorded for each pen. Sex had a significant effect on BWG and FCR (P < 0.001) where males had a significantly higher BWG and better FCR compared to females. There was a significant interaction between sex and protein level on feed intake (FI) (P < 0.05), where male birds had a higher FI compared to female birds only when the birds were fed SCP but not RCP diets. There was a significant interaction between CP level and sex on the expression of CAT2 (P = 0.02) and PEPT2 (P = 0.026) where the genes were significantly upregulated in females but only when the RCP diet was fed. The RCP diet upregulated the expression of BoAT (P = 0.03) as a main effect. Female birds had significantly higher expression of the PepT-2 gene compared to the males. The alpha diversity of the cecal microbiota showed differences among the treatments. The Shannon diversity index was statistically higher (P = 0.036) for males fed the SCP diet and the Chao1 index for evenness was statistically higher (P = 0.027) in females fed the SCP diet. There was also a difference in the relative abundance of the 15 most common genera found in the cecal content of the broilers in this experiment and lastly, the differential composition of microbiota between the different treatments was also significantly different. This study suggests that chickens are able to compensate for a reduction in AA substrates when fed a low CP diet through the upregulation of certain AA transporters, females may adapt to low CP diets better by such upregulation compared to males, and lastly, sex has an effect on the cecal microbial population and these differences contribute towards the performance differences between male and female broilers.

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.

Xylanase and beta-glucanase improve performance parameters and footpad dermatitis and modulate intestinal microbiota in broilers under an Eimeria challenge.

Coccidiosis is an enteric disease of poultry worldwide that compromises gut health and growth performance. The current research investigated the effects of 2 doses of a multienzyme preparation on broilers' performance, gut health, and footpad dermatitis (FPD) under an Eimeria challenge. A total of 512 mixed-sex day-old chicks (Cobb 500) were randomly allocated to 4 treatments of 8 replicates. Treatments were: 1) nonchallenged control (NC); 2) NC + Eimeria challenge (CC); 3) CC + recommended level of xylanase and glucanase (XG, 100 g/t feed [on top]); 4) CC + double XG (2XG, 200 g/t feed). Eimeria spp. vaccine strains were gavaged on d 9 to induce coccidiosis in chickens. Performance parameters were evaluated during starter, grower, and finisher phases, and 4 birds per pen were euthanized on d 16 for sampling, FPD was scored on d 35, and litter moisture was analyzed on d 17 and 35. The data were analyzed using 1-way ANOVA with Tukey's test to separate means, and Kruskal-Wallis test was used for non-normally distributed parameters. The results showed that the Eimeria challenge was successful based on reduced weight gain and feed intake during grower phase, and higher FITC-d concentration, lesion score (female), and oocyst counts (d 14) in CC group compared to N.C. group, while XG and 2XG increased (P < 0.001) weight gain and improved FCR compared to CC and NC groups during finisher phase. The addition of X.G. and 2XG decreased litter moisture (P = 0.003) and FPD (P < 0.001) in challenged broilers compared to the N.C. group (d 35). Supplementing XG and 2XG reestablished the population of Lactobacillus in the cecum of challenged birds to an intermediate level between the NC and CC groups (P > 0.05). The inclusion of XG tended to increase the expression of Junctional adhesion molecule 2 (JAM2), which was not different from CC and NC groups (P > 0.05). In conclusion, the combination of xylanase and glucanase (Natugrain TS) improved the performance and modulated jejunal microbiota of broilers under mild Eimeria challenge.

Buffered formic acid and a monoglyceride blend improve performance and modulate gut bacteria and immunity gene expression in broilers under necrotic enteritis challenge.

Due to the removal of antibiotics from animal feed, alternatives have been sought to control necrotic enteritis (NE) in broilers. The current study investigated the effects of buffered formic acid (Amasil NA) and monoglycerides of short- and medium-chain fatty acids (Balangut LS P) on the performance and gut health of broilers challenged with NE. A total of 816 as-hatched 1-d-old chicks (Cobb 500) were randomly assigned to 6 treatments with 8 replicates. Treatments were: T1) nonchallenged control; T2) NE challenged control; T3) Amasil NA (challenge plus Amasil NA, 0.3% throughout all phases); T4) Balangut LS P (challenge plus Balangut LS P, 0.5%, 0.3%, and 0.2% in the starter, grower and finisher phases, respectively; T5) Combined (challenge plus combination of T3 and T4); T6) Antibiotic (challenge plus Zn bacitracin, 0.05 % throughout all phases). Birds were orally gavaged with live Eimeria vaccine species (d 9) and with Clostridium perfringens (d 14 and 15). On d 16, birds were sampled to evaluate gut permeability, microbiota, and mRNA abundance in the jejunum. The data were analyzed in JMP software using one-way ANOVA with Tukey's test to separate means, and Kruskal-Wallis test was used for non-normally-distributed parameters. Results showed that Balangut LS P decreased (P<0.05) feed conversion ratio compared to nonchallenged ones at the end of the study. Balangut LS P reduced (P < 0.05) the level of cecal Bacteriods compared to nonchallenged group, whereas Amasil NA shifted the levels of ileal Bifidobacteria, Enterobacteriaceae, and Lactobacillus towards nonchallenged control (P > 0.05). NE challenge upregulated (P < 0.001) the expression of IL-21R, zeta chain of T cell receptor (ZAP70), and dual specificity phosphatase 4 (DUSP4) compared to nonchallenged birds, whereas Balangut LS P showed an intermediate (P > 0.05) expression pattern of these genes towards nonchallenged and antibiotic groups. In conclusion, combination of Balangut LS P and Amasil NA has the potential to be used as an additive to improve the performance and gut health of broiler chickens, especially under challenging conditions such as NE infections.

Sex proportion as a covariate increases the statistical test power in growth performance based experiments using as-hatched broilers.

The availability of sexed day-old broiler chicks is becoming an issue as feather sexing is no longer possible. This has great implications for broiler researchers as the use of randomly distributed mixed-sex birds may result in a greater between-pen variation and thus less statistical power than the use of single-sex birds. The objective of this study was to evaluate the effect of including sex proportion as a covariate in an analysis of covariance (ANCOVA) on the statistical power compared to analysis of variance (ANOVA) where sex was not considered. The statistical parameters examined include mean square error (MSE), the F-statistic, model fit, model significance and observed power. A total of 4 separate experiments that used mixed-sex broilers with unequal numbers of male and female birds per pen were conducted during which performance of the birds was measured. The male % in each pen was recorded during each experiment and corrected for mortality. The performance results were analysed by ANOVA and the statistical parameters were then compared to ANCOVA where sex proportion was included as a covariate. The results showed that a set of assumptions first needed to be met to run ANCOVA. In addition, if the ANOVA results show a high level of model significance and power, then ANCOVA may not be necessary. In other circumstances where the assumptions are met and model significance and observed power are low, the inclusion of sex proportion as a covariate in the analysis will help to reduce MSE, increase the F-statistic value and improve the model significance, model fit and observed power. Therefore, it is suggested that sex proportion should be considered as a covariate in ANCOVA to improve statistical power in nutritional experiments when male and female broilers are unequally and randomly distributed in pens.

Influence of sex and rearing method on performance and flock uniformity in broilers-implications for research settings.

Male and female broiler chickens differ in their growth performance, carcass part weights and nutrient requirements. The potential reasons for these differences have been explored by looking at differences in nutrient digestibility, nutrient transporter gene expression as well as gut microbiota populations between male and female birds. Studies have shown that male broilers have higher crude protein requirements compared to female broilers. The expression of monosaccharide and amino acid transporters show conflicting results as expression depends on the interactions between sex and bird age and breed as well as which tissue is sampled. Differences in microbiota populations between the genders were reported which may contribute towards performance differences, however research in this area is limited. The differences observed between the sexes contribute to increased variation in nutrition trials, and the potential to rear birds as equally mixed-sex becomes an option to reduce the variation introduced by the sex effect. Difference in rearing options obviously would only be feasible provided a quick, practical and cost-effective method of sexing birds is available, a topic that is also discussed in this review.

Rearing broilers as mixed or single-sex: relevance to performance, coefficient of variation, and flock uniformity.

With known variation in performance between male and female broilers and the fact that sourcing single-sex birds for use in research is becoming increasingly difficult, it becomes important to determine the effect of rearing method with male and female broilers on between-pen variation and body weight (BW) uniformity. We evaluated the performance response of broilers reared as single or mixed-sex to standard and reduced crude protein (CP) diets. The study was designed as a 2 × 3 factorial arrangement of treatments consisting of 672 Cobb-500 broilers assigned to 48 floor pens with 6 treatments, 8 replicates, and 14 birds per pen. The factors were rearing method (male single-sex, female single-sex, or equally mixed-sex) and dietary CP level (standard or reduced). For the overall period of the trial (d 0-35) there was a significant effect (P < 0.001) of rearing method and CP level on feed intake (FI) and feed conversion ratio (FCR). There was also a significant interaction between rearing method and CP level for BWG during d 0 to 35 (P < 0.01). There was a significant interaction between CP level and sex on d 34 BW (P < 0.01) where the reduced CP diet decreased the BW of both males and females, but to a greater extent the BW of the female birds. Dietary CP level had a significant effect on relative breast and drumstick weights with birds fed the reduced CP diet having significantly lower breast weights (P < 0.001) and higher drumstick weights (P < 0.01).This study suggests that male and female broilers have different CP requirements, and rearing birds as equally mixed-sex results in the lowest CV% for performance parameters and best BW uniformity compared to single-sex birds. Furthermore, when low CP diets are fed to broilers, they will prioritize the growth of more important body parts such as the legs.

Sources and levels of copper affect liver copper profile, intestinal morphology and cecal microbiota population of broiler chickens fed wheat-soybean meal diets.

Super dosing copper (Cu) has long been used as an alternative to antibiotic growth-promoters in broiler chickens' diet to improve gut health. This study was designed to compare nutritional and growth-promoting levels of Cu hydroxychloride (CH) with CuSO4 on gut health bio-markers and liver mineral profile of broiler chickens. Ross 308 chicks (n = 864) were randomly assigned to eight treatments, as basal diet containing no supplemental Cu; the basal diet with 15 or 200 mg/kg Cu as CuSO4; or 15, 50, 100, 150 or 200 mg/kg Cu from CH. The highest liver Cu content was observed in birds fed the diets with 200 mg/kg CuSO4 (P < 0.01). Serum FITC-d concentration as the leaky gut marker, and liver malondialdehyde concentration were not affected. Copper level or source had no effect on cecal short chain fatty acid and the mRNA expression of five jejunal genes involved in gut integrity. Negative linear responses of Cu were observed on Lactobacillus (P = 0.032), Bacteroides (P = 0.033), and Enterobacteriaceae (P = 0.028) counts. The jejunal villus height increased in birds fed CH at 200 and 100 mg/kg (P < 0.05). Increasing Cu levels, linearly and quadratically (P < 0.001), increased Cu excretion.

Potential of a mixture of eugenol and garlic tincture to improve performance and intestinal health in broilers under necrotic enteritis challenge.

Plant extracts (PE) are gaining increased attention as potential alternatives to in-feed antimicrobials (AM) due to their known antimicrobial activities. This study was conducted to examine the potential of PE, a microencapsulated product composed of eugenol and garlic tincture as an alternative to AM-agent on performance and intestinal health in broilers under necrotic enteritis (NE) challenge. A total of 960 day-old mixed-sex Cobb 500 chicks were randomly distributed to 48-floor pens with 6 treatments replicated 8 times with 20 birds each. The 6 treatments were as follows: UC, unchallenged control; CC, challenged control; PE, challenged group plus PE; AM, challenged group plus AM; FAP, challenged group plus a full dose of AM with PE; HAP, challenged group plus a half dose of AM with PE in starter, grower and finisher phases. Birds in the challenged groups were inoculated with Eimeria spp. on d 9 and Clostridium perfringens on d 14. The body weight gain (BWG), feed intake (FI), feed conversion ratio (FCR), and livability of birds were compromised, and intestinal lesions and mortality were increased (P < 0.05) by NE challenge, illustrating a successful clinical NE challenge. Birds fed AM had higher BWG and FI, and lower FCR, mortality, and intestinal lesions compared to the CC group (P < 0.05). Birds fed PE had improved FCR (P < 0.05) and livability (5.8%) in an overall period compared to the CC group. On d 16, PE supplementation reduced ileal lesion scores in only male birds (P < 0.05). Birds fed PE had decreased Eimeria maxima and Eimeria acervulina oocyst counts in caecal content (P < 0.05). Birds fed PE had decreased Escherichia brunetti and total oocyst counts in caecal content, and E. acervulina oocyst counts in ileal content in only female birds (P < 0.05). On d 35, PE supplementation reduced variation of BW in both male and female birds and increased yellowness (b∗ value, 14.4%) in the thigh. These findings suggest the potential of PE supplementation in diets to improve the performance and intestinal health of birds under clinical NE as indicated by improved FCR, livability, uniformity, reduced ileal lesions, oocyst counts and increased skin yellowness. However, the protective effect of PE may not be apparent in the presence of AM in the feed.

Organic acid blends improve intestinal integrity, modulate short-chain fatty acids profiles and alter microbiota of broilers under necrotic enteritis challenge.

Controlling enteric diseases of broilers is crucial. Among many additives, organic acids (OA) and their blends are gaining attention to combat diseases in the post-antibiotic era. The current study evaluated the potentials of short-chain fatty acids (SCFA) and medium-chain fatty acids (MCFA) blends and/or phenolic compounds on intestinal integrity, intestinal pH, caecal microbiota, and caecal SCFA profiles of broilers under necrotic enteritis (NE) challenge. The additives used were: (A) a blend of SCFA, MCFA, and a phenolic compound (SMP), (B) a blend of free and buffered SCFA with MCFA (SMF), and (C) a blend of free and buffered SCFA with a high concentration of MCFA (SHM). A total of 1,404 male parental chicks of Ross 308 broilers were randomly allocated to 78 floor pens on hatching day with 6 treatments replicated 13 times with 18 birds per pen. The treatments were: UCC, unchallenged control; CHC, challenged control; BAC, challenged group plus zinc bacitracin; SMP, challenged group plus additive SMP; SMF, challenged group plus additive SMF; SHM, challenged group plus additive SHM. Birds were challenged with field-strain Eimeria spp. on d 9 and Clostridium perfringens on d 14. Birds challenged with NE increased fluorescein isothiocyanate dextran (FITC-d) concentration in serum, reduced acetate and butyrate concentrations, and increased Bacteroides and C. perfringens load in the caeca (P < 0.05). Birds fed additives decreased FITC-d from gut to serum, reduced Bacteroides (d 16, P < 0.05) and numerically reduced C. perfringens load compared to CHC group. Birds fed additive SHM had higher concentrations of acetate and butyrate (d 21, P < 0.05) than CHC group but were not different from SMP and SMF groups. All the additives exhibited similar intestinal protection against NE compared to the BAC group indicated by FITC-d concentration in serum, acetate, propionate and butyrate concentrations in the caeca, and caecal bacterial loads except for the C. perfringens (P > 0.05). The SMP group had a higher load compared to BAC (P < 0.05). These findings suggest the promising effects of OA blends as alternatives to BAC to ameliorate the impact of NE challenge of broilers as indicated by improved intestinal health.

Detection and Quantification of Clostridium perfringens and Eimeria spp. in Poultry Dust Using Real-Time PCR Under Experimental and Field Conditions.

Infection of poultry with Eimeria spp., the causative agent of coccidiosis, can predispose birds to necrotic enteritis (NE) caused by netB gene-positive strains of Clostridium perfringens. The detection of Eimeria spp., C. perfringens, and netB were examined in settled dust from broiler flocks under experimental and field conditions. Dust samples were collected from settle plates twice weekly from two experimental flocks inoculated with three species of pathogenic Eimeria in 9-day-old chicks, followed by netB gene-positive C. perfringens 5 days later to produce subclinical and clinical NE. A noninoculated flock was sampled weekly from day 0 and served as a control flock. An additional 227 dust samples from commercial broiler flocks were collected at the end-of-batch (6-7 wk of age; one scraped dust sample per flock). In the NE-subclinical and NE-clinical flocks, high levels of Eimeria spp. and C. perfringens were detected after inoculation followed by a gradual decline over time. In the control flock, C. perfringens and netB were detected at low levels. No significant effect of sampling location was evident on Eimeria spp., C. perfringens, and netB load within poultry houses. These results provide evidence that Eimeria spp., C. perfringens, and netB gene copies can be readily measured in poultry dust samples collected in settle plates and may provide an alternative sampling method for monitoring flock coccidiosis and NE status. Further studies are required to assess the utility for such a test in commercial flocks.

Artículo regular­Detección y cuantificación de Clostridium perfringens y Eimeria spp. en polvo de instalaciones avícolas mediante PCR en tiempo real bajo condiciones experimentales y de campo. La infección de aves comerciales con Eimeria spp., el agente causante de la coccidiosis, puede predisponer a las aves a enteritis necrótica (NE) causada por cepas de Clostridium perfringens positivas a la presencia del gene netB. La detección de Eimeria spp., C. perfringens y del gene netB se examinó en el polvo sedimentado de parvadas de pollos de engorde bajo condiciones experimentales y de campo. Se recolectaron muestras de polvo por sedimentación en placa dos veces por semana de dos parvadas experimentales inoculadas con tres especies de Eimeria patógena en pollitos de nueve días, seguidas de C. perfringens positiva al gene netB cinco días después para producir enteritis necrótica subclínica y clínica. Una parvada no inoculada se muestreó semanalmente desde el día cero y sirvió como parvada control. Se recolectaron 227 muestras adicionales de polvo de parvadas de pollos de engorde comerciales al final del lote (6 a 7 semanas de edad; una muestra de polvo por raspado por parvada). En las parvadas con enteritis necrótica subclínica y clínica, se detectaron niveles altos de Eimeria spp. y de C. perfringens después de la inoculación seguida de una disminución gradual con el tiempo. En la parvada control, C. perfringens y el gene netB se detectaron en niveles bajos. No fue evidente ningún efecto significativo de la ubicación del muestreo sobre la carga de Eimeria spp., C. perfringens y del gene netB dentro de las casetas. Estos resultados proporcionan evidencia de que las copias genéticas de Eimeria spp., C. perfringens y del gene y netB se pueden medir fácilmente en muestras de polvo de instalaciones avícolas recolectadas mediante sedimentación en placa y pueden proporcionar un método de muestreo alternativo para monitorear coccidiosis y el estado de enteritis necrótica en la parvada. Se requieren más estudios para evaluar la utilidad de tal prueba en parvadas comerciales.

A Microencapsulated Mixture of Eugenol and Garlic Tincture Supplementation Mitigates the Effect of Necrotic Enteritis on Intestinal Integrity and Increases Goblet Cells in Broilers.

This study was conducted to examine the effects of a plant extract mixture, a microencapsulated product composed of eugenol and garlic tincture (PE), on intestinal health in broilers under necrotic enteritis (NE) challenge. A total of 960 d-old mixed-sex Cobb 500 chicks were randomly distributed to 48-floor pens housing 20 birds per pen. Six treatments were applied: UC, unchallenged control; CC, challenged control; PE, challenged group plus PE; AM, challenged group plus antimicrobial (AM); FAP, challenged group plus a full dose of AM with PE; HAP, challenged group plus a half dose of AM with PE in starter, grower and finisher phases. Birds in the challenged groups were inoculated with Eimeria spp. on d 9 and Clostridiumperfringens on d 14. On d 16, the CC group had increased serum fluorescein isothiocyanate dextran (FITC-d), reduced villus surface area, goblet cell number, upregulated CLDN1, JAM2 genes and reduced microbial diversity compared to the UC group (p < 0.05). Birds fed PE had reduced FITC-d, increased goblet cell number and Bifidobacterium compared to the CC group (p < 0.05). Birds fed PE had reduced CLDN5 expression in male birds, and Bacteroides spp. in female birds than CC group (p < 0.05). These findings suggest that PE supplementation mitigates the effect of NE by improving the intestinal health of birds.

In vitro inhibitory activities of sugarcane extract on avian Eimeria sporozoites.

The current in vitro study aimed to investigate the effects of a processed sugarcane extract on the viability of avian Eimeria sporozoites. Treatments were applied to hatched sporozoites: 1) without additives (no-treatment control); 2) with ethanol; 3) with salinomycin; 4) with Polygain™. All treatments were incubated in RPMI media containing live sporozoites at 37 °C for 14 h and then the number of viable sporozoites were counted. Compared to the no-treatment control, Polygain™ decreased (P < 0.001) the counts of E. maxima, E. acervulina, E. bruneti, and E. mitis sporozoites to a level similar to salinomycin (P > 0.05). In conclusion, Polygain™ could be a potential candidate as an anticoccidial agent.

Potential of blended organic acids to improve performance and health of broilers infected with necrotic enteritis.

Organic acids (OA) and their blends have been shown to positively affect performance and health of broilers. However, the data in the literature are not consistent. This study examined the potential of blended short-chain fatty acids (SCFA) with medium-chain fatty acids (MCFA) as alternatives to antibiotic growth promoters (AGP) on performance, health and welfare of broilers infected with necrotic enteritis (NE). The additives used were: A) a blend of SCFA, MCFA, and a phenolic compound (SMP); B) a blend of free and buffered SCFA with MCFA (SMF); C) a blend of free and buffered SCFA with a high concentration of MCFA (SHM). A total of 1,404 Ross 308 one-day-old male parental chicks were randomly distributed into 78-floor pens with 13 replicates of 18 birds each. Six treatments were the following: T1, unchallenged control (UCC); T2, challenged control (CHC); T3, challenged group plus zinc bacitracin (BAC); T4, challenged group plus additive SMP; T5, challenged group plus additive SMF; T6, challenged group plus additive SHM. Challenged birds were gavaged with Eimeria spp. on d 9 and Clostridium perfringens EHE-NE18 on d 14. Post NE challenge and cumulatively, BWG, FCR, and nutrient digestibility of birds were compromised (P < 0.05) by NE challenge indicating a successful induction of sub-clinical NE. Additive SHM had higher BWG compared to CHC and BAC groups (P = 0.001; d 10 to 24) but not different from SMP and SMF groups (P > 0.05). All the 3 additive groups had lower FCR compared to CHC (P = 0.001; d 0 to 35), and exhibited similar jejunal lesions (d 16) compared to BAC and apparent ileal protein digestibility (d 21) compared to UCC and BAC groups (P > 0.05). Birds in additive SHM group had a higher concentration of serum IgA compared to all groups (P = 0.001) except additive SMF (P > 0.05; d 21). All the additive groups had lower footpad dermatitis and hock burns compared to CHC (P < 0.05). The findings suggest the potential of blended OA as alternatives to BAC to protect broilers from NE indicated by improved FCR, immunity, digestibility, and bird welfare.

Multi-carbohydrase enzymes improve feed energy in broiler diets containing standard or low crude protein.

This study evaluated the effect of multi-carbohydrase (MC) on energy and nitrogen (N) balance and gene expression in broilers fed diets with different crude protein (CP) contents. The study employed a 2 × 2 factorial arrangement of treatments. The factors were presence or absence of MC, and standard (SCP) or low (LCP) dietary CP concentration. A 3-phase feeding program was used, including starter (0 to 7 d), grower (8 to 17 d) and finisher (18 to 28 d) phases. The study was undertaken in closed calorimetry chambers. Each of the 4 dietary treatments was replicated 8 times in total across 2 runs, with 2 birds per replicate (n = 64). Data for energy partitioning and N balance were collected from d 25 to 28. On d 28, birds were euthanized to collect muscle and intestinal tissue samples for gene expression. The results showed that the MC increased apparent metabolizable energy (AME, P < 0.01) and net energy (NE, P < 0.05), and reduced the feed conversion ratio (FCR, P < 0.01) in all diets. The proportion of energy retained as fat per total energy retention (REf/RE) was positively correlated with feed AME and NE (r = 0.541, P < 0.01 and r = 0.665, P < 0.001, respectively), suggesting that feed energy augmented with increased fat gain. Muscle ATP synthase subunit alpha (ATP5A1W) gene expression had a positive correlation with REf/RE and feed NE (r = 0.587, P < 0.001 and r = 0.430, P < 0.05, respectively). Similarly, muscle peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1A) expression was negatively correlated with weight gain and positively correlated with FCR (r = -0.451, P < 0.05 and r = 0.359, P < 0.05, respectively). These correlations show that over-expressions of muscle genes related to energy production reduce bird performance. This study demonstrated that MC increase dietary energy utilization, regardless of dietary CP concentration. However, the energy released by the enzymes increases feed energy-to-CP ratio, meaning there is excess energy that is then deposited as body fat. This suggests that supplemental MC in broiler feeds is beneficial if diets are formulated to contain marginal energy levels.

Detection and distribution of haematophagous flies and lice on cattle farms and potential role in the transmission of Theileria orientalis.

On the Northern Tablelands of New South Wales, Australia, endemic Theileria orientalis infection of cattle has been reported on farms where no ticks have been observed, raising the question of how Theileria is transmitted in these areas. To investigate the potential role of mechanical transmission by insects, the present study investigated the seasonal dynamics of biting fly species trapped between December 2017 and May 2019 on six cattle farms in the region where the presence of Theileria was confirmed. These flies, sucking lice collected from these farms and pools of Culicoides species affecting cattle also trapped in the region were subjected to qPCR detection of T. orientalis. Eleven species from eight genera of biting flies were collected using unbaited Nzi traps. Tabanid species were present in all farms, with Dasybasis oculata (43.6 %) and D. circumdata (27.6 %) being the most abundant and widespread species. The effect of season on the mean count per trapping event was statistically significant and highest in the summer months for Lilaea fuliginosa and D. oculata, and the autumn months for D. circumdata, with no seasonal effect on the abundance of the undescribed Dasybasis spp. No biting flies were trapped during the winter months. Sucking (Linognathus vituli and Haematopinus eurysternus) and biting (Bovicola bovis) cattle lice were also collected from all farms with the latter detected in only one farm. PCR screening for T. orientalis of trapped hematophagous insects resulted in parasite detection in the tabanid and Stomoxyini flies, biting midges and sucking lice with the highest proportion of positive samples for Haematopinus eurysternus (4/4 pools) and H. irritans exigua (6/15 individuals). The detection of the parasite in these potential vectors indicates a possible role in the mechanical transmission of T. orientalis and may partly explain the ubiquitous presence of Theileria in areas where ticks are absent.

Monoglyceride Blend Reduces Mortality, Improves Nutrient Digestibility, and Intestinal Health in Broilers Subjected to Clinical Necrotic Enteritis Challenge.

This study evaluated the potential of monoglyceride blend (MG) and buffered formic acid (FA) as alternatives to antibiotics in the performance and intestinal health of broilers under clinical necrotic enteritis (NE) challenge. A total of 544 as-hatched Ross 308 broiler chicks were randomly distributed to 32-floor pens housing 17 birds per pen. The four treatments were: NC-non-additive control; ZBS-antibiotic group supplemented with zinc bacitracin and salinomycin; MG-additive MG supplementation in the starter phase only; and MGFA-additive MG in starter phase and FA in grower and finisher phases. All birds were challenged with Eimeria spp. and Clostridium perfringens. Results showed that the NC group had lower BWG and higher FCR than the ZBS group in the grower and overall period (p < 0.05). The NC group had higher NE-caused mortality (days 14 to 17) than the ZBS group (p < 0.05). Birds fed MG had lower NE-caused mortality than the NC group (p < 0.05). Birds fed MG had upregulated jejunal tight junction protein1 (TJP1) and immunoglobulin (IgG) on day 16 and improved gross energy digestibility on day 24 than the NC group (p < 0.05). These findings suggest that supplementation of MG may improve intestinal health and protect birds from clinical NE occurrence.

Buffered formic acid and a monoglyceride blend coordinately alleviate subclinical necrotic enteritis impact in broiler chickens.

The objective of this study was to evaluate the effect of 2 different doses of a partially buffered formic acid product (Amasil NA; 61% formic acid, 20.5% sodium formate), and a monoglyceride blend of short- and medium-chain fatty acids (BalanGut LS P) on necrotic enteritis (NE) infected broilers in terms of performance, intestinal microbial population and short-chain fatty acids concentrations in the gastrointestinal tract. A total of 528-day-old as hatched Ross 308 broilers were allocated to 48 pens with 11 birds in each pen. Six dietary treatments applied in the study were: T1) nonsupplemented diet (Control); T2) antibiotic supplemented diets; T3) and T4) high (Starter: 0.5%; Grower and Finisher: 0.5%) and low (Starter: 0.3%; Grower and Finisher: 0.2%) dose of Amasil NA; and groups T5) and T6) high (Starter: 0.3%; Grower and Finisher: 0.2%) and low dose (Starter: 0.3%; Grower: 0.15%; Finisher: 0.075%) of (BalanGut LS P). All birds in this study were fed starter (d 0-10), grower (d 11-24) and finisher (d 25-35) diets and challenged with NE. To induce subclinical NE, oral administrations of Eimeria oocysts (d 9) followed by inoculation of Clostridium perfringens strains (d 14 and 15) were applied. Results showed that birds fed the high dose of Amasil NA, had a higher feed conversion ratio (FCR,P < 0.05) compared to the nonsupplemented group during the starter period. Antibiotic supplementation reduced FCR during the grower (P < 0.001), finisher (P < 0.05) and overall (P < 0.001) periods of the experiment. Both levels of BalanGut LS P and low levels of Amasil NA enhanced overall FCR (P < 0.05) compared to the birds in the nonsupplemented group. Compared to the nonsupplemented group, high levels of Amasil NA and low levels of BalanGut LS P improved FCR in the finisher stage (P < 0.05). On d 16, cecum digesta of birds fed with antibiotic supplemented diets showed a significantly lower number of C. perfringens (P < 0.001) compared to the nonsupplemented and high level of BalanGut LS P group. Bacillus (P < 0.01) and Ruminococcus numbers were significantly lower in the birds fed with high level of Amasil NA (P < 0.05) compared to the antibiotic supplemented diets. High doses of Amasil NA, showed the highest propionate concentration in the cecum (P < 0.001). The study suggests that supplementation of BalanGut LS P and Amasil NA at different feeding phases may achieve optimal performance improvement in broilers under NE challenge.

Over-processed meat and bone meal and phytase effects on broilers challenged with subclinical necrotic enteritis: Part 3. Bone mineralization and litter quality.

This study was conducted to determine the effect of necrotic enteritis (NE), phytase level and meat and bone meal (MBM) processing on bone mineralization of broilers and litter quality. Ross 308 male broiler chicks (n = 768) were allotted to 48 pens with 16 birds each. There were 8 dietary treatments in a 2 × 2 × 2 factorial arrangement. Factors were NE challenge (no or yes), phytase level (500 or 5,000 FTU/kg), and MBM (as-received or over-processed). Half of the birds were challenged with field strains of Eimeria spp. at d 9 and 108 CFU per mL of Clostridium perfringens strain EHE-NE18 on d 14 and 15. The middle toe, tibia and femur of 2 birds per pen were excised at d 16 and 29 for determination of ash, breaking strength (BS) and bone mineralization. At d 42, all were assessed for hock burns and litter was scored and assessed for dry matter (DM). At d 16, challenged birds had lower toe ash (P < 0.01), femur ash (P < 0.001), tibia ash (P < 0.001) and tibial BS (P < 0.001) than unchallenged birds. At d 16, challenged birds fed high phytase and over-processed MBM had higher toe Mn than those fed low phytase and as-received MBM. At d 29 unchallenged birds fed high phytase and as-received MBM had a higher toe Mn than those fed over-processed MBM. At d 16, a phytase × MBM interaction was detected for femur Zn concentration (P < 0.05), where a higher level of Zn was observed in the high phytase group fed over-processed MBM. At d 16, tibial Ca (P < 0.05) and P (P < 0.05) were lower in the challenged whereas the femur K (P < 0.001), Mn (P < 0.01) and Na (P < 0.001) were higher in the challenged at d 16. At d 42, challenged birds had higher litter DM (P = 0.058) and fewer hock burns than those unchallenged (P < 0.05). In conclusion, NE impaired bone traits but high phytase and over-processed MBM increased bone mineral contents. Cases of hock burns may be lower under NE incidences due to lower livability of birds reducing litter wetness.