Supplementation of lactobacillus-fermented rapeseed meal in broiler diet reduces Campylobacter jejuni cecal colonization and limits the l-tryptophan and l-histidine biosynthesis pathways.

BACKGROUND: Campylobacter jejuni (C. jejuni), a widely distributed global foodborne pathogen, primarily linked with contaminated chicken meat, poses a significant health risk. Reducing the abundance of this pathogen in poultry meat is challenging but essential. This study assessed the impact of Lactobacillus-fermented rapeseed meal (LFRM) on broilers exposed to C. jejuni-contaminated litter, evaluating growth performance, Campylobacter levels, and metagenomic profile. RESULTS: By day 35, the litter contamination successfully colonized broilers with Campylobacter spp., particularly C. jejuni. In the grower phase, LFRM improved (P < 0.05) body weight and daily weight gain, resulting in a 9.2% better feed conversion ratio during the pre-challenge period (the period before artificial infection; days 13-20). The LFRM also reduced the C. jejuni concentration in the ceca (P < 0.05), without altering alpha and beta diversity. However, metagenomic data analysis revealed LFRM targeted a reduction in the abundance of C. jejuni biosynthetic pathways of l-tryptophan and l-histidine and gene families associated with transcription and virulence factors while also possibly leading to selected stress-induced resistance mechanisms. CONCLUSION: The study demonstrated that LFRM inclusion improved growth and decreased cecal Campylobacter spp. concentration and the relative abundance of pivotal C. jejuni genes. Performance benefits likely resulted from LFRM metabolites. At the molecular level, LFRM may have reduced C. jejuni colonization, likely by decreasing the abundance of energy transduction and l-histidine and l-tryptophan biosynthesis genes otherwise required for bacterial survival and increased virulence. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Impact of precursor-derived peracetic acid on post-weaning diarrhea, intestinal microbiota, and predicted microbial functional genes in weaned pigs.

Post-weaning diarrhea affects piglets in the nursery phase of production, leading to a substantial impact both at the farm and financial levels. The multifactorial etiology of this disease includes housing conditions, pig genetics, microbial composition, and metagenomic assets. Among the common therapeutic approaches, the widely used zinc oxide underwent a European Union ban in 2022 due to its negative environmental impact and correlation to increased antimicrobial resistance. During this study, we have tested two levels of inclusion of the potential antimicrobial alternative peracetic acid, delivered in water via the hydrolysis of the precursors sodium percarbonate and tetraacetylethylenediamine, in comparison to zinc oxide and an untreated control during a 2-week animal study. We assessed the microbial composition and predicted the metagenome, together with performance and physiological parameters, in order to describe the microbial functional role in etiopathology. Both zinc oxide and peracetic acid resulted in amelioration of the diarrheal status by the end of the trial period, with noticeable zinc oxide effects visible from the first week. This was accompanied by improved performance when compared to the first-week figures and a decreased stomach pH in both peracetic acid levels. A significant reduction in both stomach and caecal Proteobacteria was recorded in the zinc oxide group, and a significant reduction of Campylobacter in the stomach was reported for both zinc oxide and one of the peracetic acid concentrations. Among other functional differences, we found that the predicted ortholog for the zonula occludens toxin, a virulence factor present in pathogens like Escherichia coli and Campylobacter jejuni, was less abundant in the stomach of treated pigs compared to the control group. In water, peracetic acid delivered via precursor hydrolysis has the potential to be a valid intervention, an alternative to antimicrobial, to assist the weaning of piglets. Our findings support the view that post-weaning diarrhea is a complex multifactorial disease with an important metagenomic component characterized by the differential abundance of specific predicted orthologs and microbial genera in the stomach and caecum of pigs.

Estimating the contribution of the porcine fecal core microbiota to metabolite production via mathematical modeling and in vitro fermentation.

The swine gut microbiota is a complex ecosystem found throughout the gastrointestinal tract, with multiple exchanges with the host and whose composition is linked to both external and internal factors, such as diet or breed. Diet, probiotic, or prebiotic interventions have been designed to boost beneficial host-microbiota interactions, such as the production of anti-inflammatory molecules, or the fermentation of otherwise undigested resources. In parallel, a smaller microbial population, shared among the same host species, independent of external or internal factors, has been described and defined as the "core microbiota." Therapies targeting the core microbiota could possibly lead to more precise and long-lasting effects. However, the metabolic role of the porcine core microbiota, especially in relation to the rest of the microbial community, is currently missing. We present here the first dynamic model of the porcine core microbiota, which we used to estimate the core-microbiota metabolite production and to forecast the effect of a synbiotic intervention targeting the core genera of the core microbiota. We developed a community model in which a total of 17 microbial groups were established based on culture-based information of representative species. First, the model parameters were estimated, and the resulting model simulations were compared favorably with in vitro experimentation. The model was then used to predict the microbial dynamics of the core and non-core members under different experimental conditions. Therefore, it was able to theorize the main-metabolite core microbiota contribution, hypothesizing that it could be mainly responsible for acetate and propionate, but not for butyrate production.IMPORTANCECurrently, little information is present in the literature to describe the generic metabolic role of the porcine core microbiota or to inform on the effect of interventions targeting the core genera. Moreover, both in vitro and in vivo experimentations aiming to explore the core microbiota dynamics are technically demanding, expensive, or restricted by ethical considerations. Modeling approaches can be used as an initial exploratory tool to develop hypotheses for targeted experimentation. Our mathematical model provides initial information on the microbial and metabolite dynamics of the core microbiota in relation to diet and therapeutic intervention.

Encapsulated peracetic acid as a valid broad-spectrum antimicrobial alternative, leading to beneficial microbiota compositional changes and enhanced performance in broiler chickens.

BACKGROUND: Antimicrobial alternatives are urgently needed, including for poultry production systems. In this study, we tested the potential broad-range antimicrobial alternative peracetic acid, delivered in feed via the hydrolysis of encapsulated precursors through a 28-day study using 375 Ross 308 broiler chickens. We tested two peracetic acid concentrations, 30 and 80 mg/kg on birds housed on re-used litter, and we evaluated the impact of both levels on gut microbial communities, bacterial concentration, antimicrobial resistance genes relative abundance and growth performance when compared to control birds housed on either clean or re-used litter. RESULTS: Body weight gain and feed conversion ratio improved in peracetic acid fed birds. At d 28, birds given 30 mg/kg of peracetic acid had a decreased Firmicutes and an increased Proteobacteria abundance in the jejunum, accompanied by an increase in Bacillus, Flavonifractor and Rombustia in the caeca, and a decreased abundance of tetracycline resistance genes. Chicken given 80 mg/kg of peracetic acid had greater caecal abundance of macrolides lincosamides and streptogramins resistance genes. Growth performance on clean litter was reduced compared to re-used litter, which concurred with increased caecal abundance of Blautia, decreased caecal abundance of Escherichia/Shigella, Anaerostipes and Jeotgalicoccus, and greater gene abundance of vancomycin, tetracycline, and macrolides resistance genes. CONCLUSIONS: Peracetic acid could be used as a safe broad-spectrum antimicrobial alternative in broilers. Encapsulated precursors were able to reduce the bacterial concentration in the jejunum whilst promoting the proliferation of probiotic genera in the caeca, especially at the low peracetic acid concentrations tested, and improve growth performance. Moreover, our findings offer further insights on potential benefits of rearing birds on re-used litter, suggesting that the latter could be associated with better performance and reduced antimicrobial resistance risk compared to clean litter rearing.

Growth performance, caecal microbiome profile, short-chain fatty acids, and litter characteristics in response to placement on reused litter and combined threonine, arginine and glutamine supplementation to juvenile male broiler chickens.

BACKGROUND: Exposure of broilers to litter microbiome may increase specific amino acid (AA) requirements towards activated immune responses. This may challenge the generality of the ideal protein (IP) concept, in which dietary essential AA to lysine ratios aimed to mimic presumably constant AA to lysine ratios in whole bird requirements. Therefore, we tested the effect of threonine, arginine and glutamine (TAG) supplementation to IP-based control diets (C) on performance, caecal microbiome composition, short-chain fatty acids and litter characteristics of broiler chickens placed on reused litter. RESULTS: Thirty-two pens with ten male broiler chickens each were used in a 2 × 2 factorial arrangement of two diet treatments (with or without TAG supplementation) and two litter treatments (placement on clean or reused litter) for 21 days (n = 8). Caecal contents were analysed for microbiome profile using percent guanine + cytosine (%G + C profile) method and short chain fatty acids. TAG-supplemented birds underperformed compared to C birds (P = 0.002), whereas birds placed on reused litter outperformed those on clean litter (P = 0.047). Diet, reused litter and their interaction impacted the %G + C profile at different ranges. Whilst TAG supplementation reduced bacterial abundance at %G + C 51-56 (P < 0.05), reused litter placement tended to reduce %G + C 23-31 and increase %G + C 56-59 (P < 0.10). However, TAG supplementation reduced bacterial abundance at %G + C 47-51 (P < 0.05) and increased caecal branched chain fatty acids on clean litter only (P = 0.025). Greater levels of propionic acid were observed for C birds placed on reused litter only (P = 0.008). Litter pH was greater for reused litter pens than clean litter pens at day 21 (P < 0.001). In addition, litter moisture content was less for TAG birds and reused litter pens compared to C birds (P = 0.041) and clean litter pens (P < 0.001), respectively. CONCLUSIONS: These data support the view that irrespective of performance benefits arising from bird placement on reused litter, TAG supplementation to IP-formulated baseline rations impaired growth, supported by the lowered abundance of caecal bacteria known to dominate in well-performing birds and greater levels of caecal branched chain fatty acids.

Precursor-derived in-water peracetic acid impacts on broiler performance, gut microbiota, and antimicrobial resistance genes.

Past antimicrobial misuse has led to the spread of antimicrobial resistance amongst pathogens, reportedly a major public health threat. Attempts to reduce the spread of antimicrobial resistant (AMR) bacteria are in place worldwide, among which finding alternatives to antimicrobials have a pivotal role. Such molecules could be used as "green alternatives" to reduce the bacterial load either by targeting specific bacterial groups or more generically, functioning as biocides when delivered in vivo. In this study, the effect of in-water peracetic acid as a broad-spectrum antibiotic alternative for broilers was assessed via hydrolysis of precursors sodium percarbonate and tetraacetylethylenediamine. Six equidistant peracetic acid levels were tested from 0 to 50 ppm using four pens per treatment and 4 birds per pen (i.e., 16 birds per treatment and 96 in total). Peracetic acid was administered daily from d 7 to 14 of age whilst measuring performance parameters and end-point bacterial concentration (qPCR) in crop, jejunum, and ceca, as well as crop 16S sequencing. PAA treatment, especially at 20, 30, and 40 ppm, increased body weight at d 14, and feed intake during PAA exposure compared to control (P < 0.05). PAA decreased bacterial concentration in the crop only (P < 0.05), which was correlated to better performance (P < 0.05). Although no differences in alpha- and beta-diversity were found, it was observed a reduction of Lactobacillus (P < 0.05) and Flectobacillus (P < 0.05) in most treatments compared to control, together with an increased abundance of predicted 4-aminobutanoate degradation (V) pathway. The analysis of the AMR genes did not point towards any systematic differences in gene abundance due to treatment administration. This, together with the rest of our observations could indicate that proximal gut microbiota modulation could result in performance amelioration. Thus, peracetic acid may be a valid antimicrobial alternative that could also positively affect performance.

Recent Progress in Phage Therapy to Modulate Multidrug-Resistant Acinetobacter baumannii, including in Human and Poultry.

Acinetobacter baumannii is a multidrug-resistant and invasive pathogen associated with the etiopathology of both an increasing number of nosocomial infections and is of relevance to poultry production systems. Multidrug-resistant Acinetobacter baumannii has been reported in connection to severe challenges to clinical treatment, mostly due to an increased rate of resistance to carbapenems. Amid the possible strategies aiming to reduce the insurgence of antimicrobial resistance, phage therapy has gained particular importance for the treatment of bacterial infections. This review summarizes the different phage-therapy approaches currently in use for multiple-drug resistant Acinetobacter baumannii, including single phage therapy, phage cocktails, phage-antibiotic combination therapy, phage-derived enzymes active on Acinetobacter baumannii and some novel technologies based on phage interventions. Although phage therapy represents a potential treatment solution for multidrug-resistant Acinetobacter baumannii, further research is needed to unravel some unanswered questions, especially in regard to its in vivo applications, before possible routine clinical use.

Bacterial concentration and Campylobacter spp. quantification differ when fresh or ultra-frozen samples are analysed over time using molecular biology and culture-based methods.

The study aimed to delineate the robustness of the culture-based and molecular biology methods to assess the total bacterial concentration and Campylobacter jejuni (C. jejuni) quantification in caecal content, analysed as fresh or after being stored immediately at ultra-low (-80°C) temperature at different time points (for 3, 7, 14, 28 and 62 days post collection). The caecal content was collected from birds that were artificially colonised with C. jejuni (in-vivo), and quantification was performed using both colony-forming unit (CFU) and qPCR. The results showed that storage time affected the output of culture-based analyses but mostly did not alter concentration retrieved via qPCR. After an initial ~4.5 log10 reduction in CFU observed from fresh (day 0) to frozen samples, bacterial concentration retrieved with culture-based methods seemed to be constant in samples frozen for 3 to 62 days, indicating a possible threshold for C. jejuni loss of viability due to effect of storage temperature. Ranking order analyses, revealed that the molecular biology technique was able to attribute somewhat the same relative C. jejuni concentrations to the samples analysed via qPCR. However, day 0 measurements from culture-based methods were associated with the absence of or negatively weak correlations with the rest of the time points, but ranking order was maintained from day 3 onwards. On the other hand, ranking order correlations were less constant when measuring total bacterial concentration through qPCR. The study suggests that if biological samples can't be analysed as fresh (immediately after collection) and have to be stored prior to analysis, then storage at -80°C samples be recommended to avoid the temporal-dependent effects on C. jejuni concentrations. In addition, irrespective of the method of analysis, an initial loss of CFU must be factored in when interpreting the results obtained from frozen samples.

Dietary protein supplementation results in molecular and cellular changes related to T helper type 2 immunity in the lung and small intestine in lactating rats re-infected with Nippostrongylus brasiliensis.

Acquired immunity to gastrointestinal nematodes reduces during late pregnancy and lactation which is known as periparturient relaxation of immunity (PPRI). Protein supplementation reduces the degree of PPRI in a rat model re-infected with Nippostrongylus brasiliensis, but the underlying molecular mechanisms have yet to be elucidated. Here, we hypothesized that protein supplementation will enhance T helper type 2 immunity (Th2) in the lung and small intestine. Nulliparous Sprague-Dawley rats were given a primary infection of N. brasiliensis prior to mating and restrictedly fed diets with either low protein (LP) or high protein (HP) during pregnancy and lactation. Dams were secondary infected with N. brasiliensis on day 2 post-parturition, and histology and gene expression were analysed for tissue samples collected at days 5, 8 and 11. Genes related to Th2 immunity in the lung, Retnla, Il13 and Mmp12, and in the intestine, Retnlb, were upregulated in HP dams compared to LP dams, which indicates the effect of dietary protein on Th2 immunity. HP dams also had increased splenic CD68+ macrophage populations compared to LP dams following secondary infection, suggesting enhanced immunity at a cellular level. Our data assist to define strategic utilization of nutrient supply in mammals undergoing reproductive and lactational efforts.

Temporal and nutritional effects on the weaner pig ileal microbiota.

BACKGROUND: The porcine gastrointestinal microbiota has been linked to both host health and performance. Most pig gut microbiota studies target faecal material, which is not representative of microbiota dynamics in other discrete gut sections. The weaning transition period in pigs is a key development stage, with gastrointestinal problems being prominent after often sudden introduction to a solid diet. A better understanding of both temporal and nutritional effects on the small intestinal microbiota is required. Here, the development of the porcine ileal microbiota under differing levels of dietary protein was observed over the immediate post-weaning period. RESULTS: Ileal digesta samples were obtained at post-mortem prior to weaning day (day - 1) for baseline measurements. The remaining pigs were introduced to either an 18% (low) or 23% (high) protein diet on weaning day (day 0) and further ileal digesta sampling was carried out at days 5, 9 and 13 post-weaning. We identified significant changes in microbiome structure (P = 0.01), a reduction in microbiome richness (P = 0.02) and changes in the abundance of specific bacterial taxa from baseline until 13 days post-weaning. The ileal microbiota became less stable after the introduction to a solid diet at weaning (P = 0.036), was highly variable between pigs and no relationship was observed between average daily weight gain and microbiota composition. The ileal microbiota was less stable in pigs fed the high protein diet (P = 0.05), with several pathogenic bacterial genera being significantly higher in abundance in this group. Samples from the low protein and high protein groups did not cluster separately by their CAZyme (carbohydrate-active enzyme) composition, but GH33 exosialidases were found to be significantly more abundant in the HP group (P = 0.006). CONCLUSIONS: The weaner pig ileal microbiota changed rapidly and was initially destabilised by the sudden introduction to feed. Nutritional composition influenced ileal microbiota development, with the high protein diet being associated with an increased abundance of significant porcine pathogens and the upregulation of GH33 exosialidases-which can influence host-microbe interactions and pathogenicity. These findings contribute to our understanding of a lesser studied gut compartment that is not only a key site of digestion, but also a target for the development of nutritional interventions to improve gut health and host growth performance during the critical weaning transition period.

Impact of reduced dietary crude protein levels and phytase enzyme supplementation on growth response, slurry characteristics, and gas emissions of growing pigs.

This experiment was carried out to evaluate the effect of reduced dietary crude protein (CP) levels supplemented with or without exogenous phytase on growing pigs. Six dietary treatments arranged in a 3 × 2 factorial arrangements of 3 CP levels (containing 14%, 16%, and 18% CP) supplemented each with or without 5,000 FTU/g phytase enzyme. Thirty growing pigs (average weight of 17.80 ± 0.10 kg) were allotted to the six dietary treatments in a complete randomized design. The final weight, daily weight gain, and feed conversion ratio (FCR) increased significantly with increasing CP levels. While, phytase supplementation improved (p = .044) FCR in pigs. Total solid and volatile solid content of the slurry were higher (p = .001) in pigs fed 14% and 16% CP diets supplemented with phytase when compared with other treatment groups. Concentration of methane gas emitted was lowest (p = .001) in the slurry of pigs fed 14% CP diet with or without phytase and those fed 16% CP diet with phytase supplementation. In conclusion, reduction in dietary CP levels resulted in reduced weight gain and poor FCR. While, reduced CP with phytase supplementation reduced concentration of methane gas emitted.

Changes in the Ileal, but Not Fecal, Microbiome in Response to Increased Dietary Protein Level and Enterotoxigenic Escherichia coli Exposure in Pigs.

The relationship between porcine gut microbiota composition and health is an important area of research, especially due to the need to find alternatives to antimicrobial use to manage disease in livestock production systems. Previous work has indicated that lower crude dietary protein levels can reduce the impacts of postweaning colibacillosis, which is a porcine diarrheal disease caused by enterotoxigenic Escherichia coli (ETEC). Here, to explore the complex interactions between the gut microbiota, protein nutrition, and ETEC exposure, the microbial compositions of both ileal digesta and feces were analyzed with or without ETEC exposure from pigs fed a low- or high-protein diet. Since ETEC colonization is mostly localized to the ileum, changes in the small intestinal microbiota were expected in response to ETEC exposure. This was supported by the study findings, which identified significant microbiota changes in ileal samples but not in fecal samples. Both increased dietary protein and ETEC exposure impacted on ileal microbiota alpha diversity (richness and diversity indices) and beta diversity (structure, stability, and relative taxon abundances) at certain sampling points, although the combination of a high-protein diet and ETEC exposure had the most profound impact on ileal microbiota composition. An understanding of how infection and nutrition lead to microbiota changes is likely to be required if dietary strategies are to be developed for the management of enteric diseases.IMPORTANCE Gut bacterial communities have been shown to play a key role in pig health and development and are strongly influenced by host diet, but studies highlighting the complex interactions between nutrition, gut infections and the microbiome tend to focus on bacterial populations in the feces and not other important gut locations. We found that alteration of dietary protein level and exposure to a pathogenic microorganism, enterotoxigenic Escherichia coli (ETEC), changed bacterial populations in the distal small intestine (i.e., the ileum). We found that the most profound changes occurred in pigs fed a high-protein diet in combination with exposure to ETEC, showing a clear interaction between dietary composition and exposure to a key pathogen. These changes were not observed in the fecal samples, revealing the importance of studying biologically pertinent sites in the gut, and so the data will help to inform the development of alternative management strategies for enteric disorders.

Evaluation of the nutritive value of legume alternatives to soybean meal for broiler chickens.

Five experiments were conducted to investigate the nutritional value of various legumes and a faba beans processing co-product for broilers. In Expt. 1 and 3, metabolizable energy (AME) content was evaluated for 2 batches of bean starch concentrate (BSC) that differed in physical and chemical characteristics. Standardized ileal amino acid digestibility (SIAAD) was determined for BSC in Expt. 2, and for corn, soybean meal (SBM), organic and conventional faba beans, and quinoa (Expt. 4). The growth performance response of broiler chickens to partial replacement of wheat and SBM with various legumes was investigated in Expt. 5. The AME of the BSC assayed in Expt. 1 was lower (P < 0.01) than that of the BSC assayed in Expt. 3. The SIAAD was generally high for BSC in Expt. 2 although the content and digestibility of sulfur amino acids were low. In Expt. 4, there was no difference in SIAAD of Arg, Phe, Asp, and Gly among the different feedstuffs assayed. SIAAD was largely similar for both conventional and organic faba bean. The SIAADs of Met, Thr, Ser, and Tyr were lower (P < 0.05) for quinoa compared with SBM or corn. In Expt. 5, FCR was greater (P < 0.05) for broiler chickens receiving faba beans+barley mix or lupins compared with the wheat-SBM control diet. Amino acid digestibility was greater (P < 0.01) for the diets containing lupins compared with the other diets except for Lys, Met, Thr, Ala, Asp, and Ser. On the other hand, amino acid digestibility in diet with faba beans+barley mix was lower (P < 0.05) compared with all the other diets, except for Arg, Asp, Lys, and Thr. It was concluded from the current studies that there is scope for using the assayed legumes, co-products, and quinoa in broiler chickens to partly replace SBM as protein feedstuffs.

Body Protein Reserves Sustain Maternal Performance in Early Lactation but Dietary Protein Is Necessary to Maintain Performance and Immune Responses to Nippostrongylus brasiliensis in Lactating Rats.

Background: It has been shown that dietary protein supplementation during lactation boosts immunity in Nippostrongylus brasiliensis-infected periparturient rats. It is not known whether body protein reserves accumulated during gestation have a similar effect during lactation. Objective: This study aimed to quantify the impact of body protein reserves and dietary protein supplementation on maternal performance and immune responses to N. brasiliensis during lactation. Methods: Multiparous female Sprague-Dawley rats were administered a primary infection of N. brasiliensis before mating and were restriction-fed either 60 g [low-protein diet gestation (Lge)] or 210 g [high-protein diet gestation (Hge)] crude protein (CP) per kilogram of dry matter (DM) until parturition. From parturition onward, dams were restriction-fed either 100 g [low-protein diet lactation (Lla)] or 300 g [high-protein diet lactation (Hla)] CP per kilogram of DM, generating 4 different dietary treatments. A subset of rats was sampled before parturition; postparturition, dams were secondarily infected with N. brasiliensis and samples were collected at days 5 and 11 postparturition. Results: Maternal performance until parturition, as measured by pup weight, was better in Hge rats than in Lge rats [Lge: 4.84 g; Hge: 6.15 g; standard error of the difference (SED): 0.19]. On day 11, pup weights of dams with reduced protein reserves fed protein during lactation (Lge-Hla; 20.28 g) were higher than their counterparts from Hge-Lla dams (17.88 g; SED: 0.92). Worm counts were significantly different between Lge-Lla-fed (253; 95% CI: 124, 382) and Hge-Hla-fed (87; 95% CI: 22, 104) dams on day 11 (P = 0.024). The expression of splenic interleukin 13 (Il13) and arachidonate 15-lipoxygenase (Alox15) was significantly higher (P < 0.05) in Hge-Hla dams compared with Lge-Lla dams on day 5. Conclusions: Although protein reserves were adequate to maintain maternal performance in the early stage of lactation in dams infected with N. brasiliensis, they were not adequate to maintain maternal performance and effective immune responses at later stages. Dietary protein supplementation was required to achieve this.

Analysis of temporal fecal microbiota dynamics in weaner pigs with and without exposure to enterotoxigenic Escherichia coli1,2.

The primary aim of this work was to study potential effects of subclinical enterotoxigenic Escherichia coli (ETEC) exposure on porcine fecal microbiota composition, with a secondary aim of profiling temporal shifts in bacterial communities over the weaning transition period. 16S rRNA gene metabarcoding and quantitative PCR (qPCR) were used to profile the fecal microbiota and quantify ETEC excretion in the feces, respectively. Temporal shifts in fecal microbiota structure and stability were observed across the immediate postweaning period (P < 0.05), including significant shifts in the relative levels of specific bacterial phylotypes (P < 0.05). ETEC exposure did not change the fecal microbiota structure (P > 0.05), but significant variations in fecal community structure and stability were linked to variations in ETEC excretion level at particular time points (P < 0.05). In this study, marked temporal changes in microbiota structure and stability were evident over the short weaning transition period, with a relationship between ETEC excretion level and fecal microbiota composition being observed. This study has provided a detailed analysis of fecal microbiota dynamics in the pig, which should help to inform the development of novel management strategies for enteric disorders based on an improved understanding of microbial populations during the challenging postweaning period.

Tolerance and resistance to a nematode challenge are not always mutually exclusive.

The relationship between the manifestations of tolerance (a host's ability to reduce the impact of a given level of pathogens) and resistance (a host's ability to clear pathogens) has been assumed to be an antagonistic one. Here we tested the hypothesis that mice from strains more resistant to intestinal nematodes will experience reduced tolerance compared with less resistant mice. Three inbred strains of mice were used: C57BL/6 mice have been characterised as susceptible, whereas BALB/c and NIH mice have been characterised as resistant to Heligmosomoides bakeri infection. Mice of each strain were either parasitised with a single dose of 250 L3H. bakeri (n=10) in water or were sham-infected with water (n=10). Body weight, food intake and worm egg output were recorded regularly throughout the experiment. Forty-two days p.i. mice were euthanised and organ weights, eggs in colon and worm counts were determined. C57BL/6 mice showed significantly greater worm egg output (P<0.001), eggs in colon (P<0.05) and female worm fecundity (P<0.05) compared with NIH and BALB/c mice. Parasitised BALB/c mice grew more whilst parasitised C57BL/6 mice grew less than their sham-infected counterparts during the first 2 weeks post-challenge (P=0.05). Parasitism significantly increased liver, spleen, small intestine and caecum weights (P<0.001) but reduced carcass weight (P<0.01). Average daily weight gain and worm numbers were positively correlated in NIH mice (P=0.05); however, the relationship was reversed when carcass weight was used as a measure for tolerance. BALB/c mice did not appear to suffer from the consequences of parasitism, with carcass weight similar in all animals. Our hypothesis that strains more resistant to the H. bakeri infection are less tolerant compared with less resistant strains is rejected, as the two resistant strains showed variable tolerance. Thus, tolerance and resistance to an intestinal nematode infection are not always mutually exclusive.

Ethno-medicinal study of plants used for treatment of human and livestock ailments by traditional healers in South Omo, Southern Ethiopia.

BACKGROUND: Plants have traditionally been used for treatment of human and livestock ailments in Ethiopia by different ethnic and social groups. However, this valuable source of knowledge is not adequately documented, which impedes their widespread use, evaluation and validation. Here, we recorded indigenous knowledge and standard practices for human and livestock disease control, of three ethnic groups (Aari, Maale and Bena-Tsemay) in South Omo Zone of Southern Nations, Nationalities and Peoples Regional State, Ethiopia. METHODS: A cross-sectional study was carried out using a semi-structured questionnaire to document knowledge of 50 traditional healers (40 male and 10 female) in medicinal plant use for treatment of human and livestock ailments. Descriptive statistics were used to analyze and summarize the ethno-botanical data. RESULTS: Ninety-one plants, with claimed medicinal properties against a total of 34 human and livestock ailments, were reported and botanically identified as belonging to 57 genera and 33 plant families. Most of the plant species reported belonged to one of seven major families: Lamiaceae, Solanaceae, Menispermiaceae, Fabaceae, Asteraceae, Plumbaginaceae and Geraniaceae. Woody plants (shrubs 21% and trees 29%) were the major growth form used, whilst roots (40%) and leaves (35%) were the major plant parts used in the study areas. Healers mostly practice oral administration of plant preparations (65%). Multiple medicinal plants were cited against particular ailments, and mixing of two or more different medicinal plants (14.3%) against a single ailment was also commonly reported. CONCLUSION: This study showed that traditional medicine, mainly involving the use of medicinal plants, is playing a significant role in meeting the primary healthcare needs of the three ethnic groups. Acceptance of traditional medicine and limited access to modern healthcare facilities could be considered as the main factors for the continuation of the practice. Documented knowledge of the traditional healers can be used to support the country's human and livestock health care system and improve lives and livelihoods. Information generated will be used in future studies to validate bioactivity of selected medicinal plants used by traditional healers, so to increase their acceptability in health care systems both nationally and internationally.

Leucine and methionine deficiency impairs immunity to gastrointestinal parasites during lactation.

Lactating rats reinfected with Nippostrongylus brasiliensis fed low-crude protein (CP) foods show reduced lactational performance and less resistance to parasites compared with their high-CP counterparts. Here, we hypothesised that feeding high-CP foods deficient in specific essential amino acids (AA) would result in similar penalties. Second-parity lactating rats, immunised with 1600 N. brasiliensis infective larvae before mating, were fed foods with either 250 (high protein; HP) or 150 (low protein; LP) g CP/kg, or were HP deficient in either leucine (HP-Leu) or methionine (HP-Met). On day 1 of lactation, litter size was standardised at twelve pups. On day 2, dams were either reinfected with 1600 N. brasiliensis larvae or sham-infected with PBS. Dams and litters were weighed daily until either day 8 or 11, when worm burdens, and inflammatory cells and systemic levels of N. brasiliensis-specific Ig isotypes were assessed. Data from five out of sixteen HP-Met rats were omitted due to very high levels of food refusals from parturition onwards. Relative to feeding HP foods, feeding LP, HP-Met and HP-Leu foods reduced dam weight gain and, to a lesser extent, litter weight gain, and increased the number of worm eggs in the colon, indicative of a reduction in resistance to parasites. However, only feeding LP and HP-Leu foods resulted in increased worm numbers, while none of the feeding treatments affected systemic Ig, mast and goblet cells, and eosinophil numbers. The present results support the view that resistance to parasites during lactation may be sensitive to specific essential AA scarcity.

Manipulating small ruminant parasite epidemiology through the combination of nutritional strategies.

It is increasingly being recognized that non-chemical parasite control strategies may need to be combined to control more effectively gastrointestinal parasitism, result in resilient production systems and reduce reliance on anthelmintics. Here, we consider if and how metabolizable protein (MP) supplementation and anti-parasitic plant secondary metabolites (PSM) may modulate parasite epidemiology through intervention in pasture contamination, development of infection on pasture and larval challenge as target processes. We then propose that combining two or more non-chemical parasite control strategies may have additive effects on host resistance, especially if the individual strategies target different drivers of parasite epidemiology, different processes in the parasite life cycle or different phases of acquired immunity to parasites. This epidemiological framework is used to review recent findings on combining maternal MP supplementation and grazing the PSM-rich bioactive forage chicory as an example of combining nutritional treatments to manipulate parasite epidemiology in a temperate production system. In the absence of available data for combined nutritional strategies in tropical production systems, we make predictions on the consequences of combining such strategies in these systems. We conclude that currently published studies on combining nutritional strategies under temperate conditions show potential to improve additively host resilience and reduce reliance on anthelmintics; however, effects on epidemiology have to date not shown the additive results hypothesized. The framework developed may assist further in evaluating combined (nutritional) strategies to manipulate parasite epidemiology.

Interactive effects of protein nutrition, genetic growth potential and Heligmosomoides bakeri infection pressure on resilience and resistance in mice.

The ability of animals to cope with an increasing parasite load, in terms of resilience and resistance, may be affected by both nutrient supply and demand. Here, we hypothesized that host nutrition and growth potential interact and influence the ability of mice to cope with different parasite doses. Mice selected for high (ROH) or low (ROL) body weight were fed a low (40 g/kg; LP) or high (230 g/kg; HP) protein diet and infected with 0, 50, 100, 150, 200 or 250 L3 infective Heligmosomoides bakeri larvae. ROH-LP mice grew less at doses of 150 L3 and above, whilst growth of ROH-HP and of ROL mice was not affected by infection pressure. Total worm burdens reached a plateau at doses of 150L3, whilst ROH mice excreted fewer worm eggs than ROL mice. Serum antibodies increased with infection dose and ROH mice were found to have higher parasite-specific IgG1 titres than ROL mice. In contrast, ROL had higher total IgE titres than ROH mice, only on HP diets. The interaction between host nutrition and growth potential appears to differentially affect resilience and resistance in mice. However, the results support the view that parasitism penalises performance in animals selected for higher growth.