Shifts in microbiota and gene expression of nutrient transporters, mucin and interleukins in the gut of fast-growing and slow-growing chickens infected by Salmonella Enteritidis.

Slow-growing breeds are more resistant to Salmonella infection compared to fast-growing broilers. However, it is unclear whether that is associated with innate resistance or rather rely on differences in Salmonella-induced gut responses. We investigated the microbial composition and gene expression of nutrient transporters, mucin, and interleukin in the gut of a fast-growing (Cobb500) and a slow-growing naked neck (NN) chicken breeds challenged with Salmonella Enteritidis. Hatchlings were inoculated at two days of age using sterile broth (sham) or Salmonella Enteritidis (SE) and distributed according to a completely randomized design into four treatments: Cobb-sham; Cobb-SE; NN-sham; and NN-SE. Cecal SE counting and microbial composition by 16 S rRNA sequencing were determined at 24-, 96-, and 168-hours post-inoculation (hpi). Gene expression of amino acid (Asct1) and peptide transporters (PepT1), glucose transporters (Sglt1, Glut2 and Glut5) and mucin (Muc2) in the jejunum and expression of interleukins (IL1 beta, IL8, IL17 and IL22) in the cecum was assessed by qPCR at 24 and 168 hpi. NN birds were colonized by SE just as Cobb birds but showed innate upregulation of Muc2, IL8 and IL17 in comparison to Cobb. While nutrient transporter mRNA expression was impaired in SE-challenged Cobb birds, the opposite was observed in NN. There were no differences in microbial diversity at different sampling times for Cobb-SE, whereas the other groups had higher diversity and lower dominance at 24 hpi compared with 96 hpi and 168 hpi. NN birds apparently develop earlier gut microbial stability, have higher basal level of mucin gene expression as well as differential nutrient transporter and interleukin gene expression in the presence of SE which might mitigate the effects of SE infection compared to Cobb birds.

Effect of diets containing commercial bioactive compounds on Salmonella Heidelberg infection in broiler chicks.

Salmonella Heidelberg (SH) is responsible for economic losses in poultry farming and food infections in humans and is a serious public health problem. Recently, there has been an increase in the frequency of isolation of this serotype in batches of broilers raised in Brazil. It is necessary to find new ways to help control this pathogen. The present study aimed to evaluate the effect of diets containing the compound Original XPC, which is a prebiotic-like fermented compound (PFC), and/or Sangrovit, which is a sanguinarine-based phytobiotic (SAN), on SH infection in broiler chicks. For this purpose, SH colonization in the cecum and its invasion into the spleen and liver were evaluated, as were the histopathological changes caused in these organs. The lowest cecal SH counts were observed in birds that ingested SAN, followed by those fed PFC (P < 0.05), with no added effect when the two bioproducts were used together (SAN + PFC). The mean SH and liver spleen counts did not differ between groups (P > 0.05). In general, birds from all groups challenged with SH showed similar macroscopic changes, such as hemorrhagic areas, hepatomegaly, and splenomegaly, such changes being more intense in the infected control group. The microscopic changes observed in the liver included hepatocyte congestion, heterophil infiltration in the sinusoid capillaries, areas of necrosis, and mononuclear inflammation. In the cecum, heterophilic infiltrate and thickening of the lamina propria were observed. In the ileum, the most common changes were congestion and thickening of the lamina propria and atrophy of the villi and crypts. The microscopic changes were less intense in the supplemented birds than the infected control group, and those supplemented with SAN developed the least changes. As ideal conditions for histomorphometric parameters of the ileum, the villus:crypt ratio in birds should be high, the villi should be long, and the crypts should be shallow. In the present study, higher mean heights and villus areas were observed in uninfected control and SAN group birds, and the crypt depth was lower in birds in the negative control group. The lowest villus:crypt ratio was observed in the birds of the infected control group. Although additional studies are needed, the preliminary results of the current investigation indicated that the addition of bioproducts, especially SAN, to the diet of birds helped to control SH infection, reducing its count in the cecum and improving overall and intestinal health.

Hydroxy-selenomethionine enhances the productivity and egg quality of 50- to 70-week-old semi-heavy laying hens under heat stress.

Oxidative stress significantly compromises the production efficiency of laying hens. It has been reported in literature that selenium (Se) in poultry diets has a positive effect on mitigating these effects. This study has been carried out to evaluate the effects of Se supplementation in feeds, from either an inorganic or a hydroxy-selenomethionine (OH-SeMet) source, on the performance and physiological traits of 50- to 70-wk-old Dekalb Brown laying hens under heat stress, and on their egg quality after different storage durations. The treatments consisted in supplementing 0.3 ppm of Se as sodium selenite (SS; 45%-0.7g/ton) or OH-SeMet (2%-15g/ton) in twelve 16-bird replicates. Supplementation with OH-SeMet resulted in a better performance of the laying hens than with SS: -5% feed conversion ratio and +3.6% of egg mass. A reduction in egg quality was observed with prolonged egg storage, which was mitigated with the use of OH-SeMet in laying hen diets. The use of OH-SeMet increased the antioxidant capacity of the birds, which showed higher glutathione peroxidase levels in the blood, kidneys, liver, and intestinal mucosa, in addition to a higher Se content in the eggs and a greater bone resistance. Thus, supplementing feeds with 0.3 ppm of OH-SeMet to 50- to 70-wk-old semi-heavy laying hens enhances their antioxidant capacity and leads to a higher egg quality and productivity than SS supplementation.

Revisiting the Biological Behavior of Salmonella enterica in Hydric Resources: A Meta-Analysis Study Addressing the Critical Role of Environmental Water on Food Safety and Public Health.

The increasing number of studies reporting the presence of Salmonella in environmental water sources suggests that it is beyond incidental findings originated from sparse fecal contamination events. However, there is no consensus on the occurrence of Salmonella as its relative serovar representation across non-recycled water sources. We conducted a meta-analysis of proportions by fitting a random-effects model using the restricted maximum-likelihood estimator to obtain the weighted average proportion and between-study variance associated with the occurrence of Salmonella in water sources. Moreover, meta-regression and non-parametric supervised machine learning method were performed to predict the effect of moderators on the frequency of Salmonella in non-recycled water sources. Three sequential steps (identification of information sources, screening and eligibility) were performed to obtain a preliminary selection from identified abstracts and article titles. Questions related to the frequency of Salmonella in aquatic environments, as well as putative differences in the relative frequencies of the reported Salmonella serovars and the role of potential variable moderators (sample source, country, and sample volume) were formulated according to the population, intervention, comparison, and outcome method (PICO). The results were reported according to the Preferred Reporting Items for Systematic Review and Meta-Analyzes statement (PRISMA). A total of 26 eligible papers reporting 148 different Salmonella serovars were retrieved. According to our model, the Salmonella frequency in non-recycled water sources was 0.19 [CI: 0.14; 0.25]. The source of water was identified as the most import variable affecting the frequency of Salmonella, estimated as 0.31 and 0.17% for surface and groundwater, respectively. There was a higher frequency of Salmonella in countries with lower human development index (HDI). Small volume samples of surface water resulted in lower detectable Salmonella frequencies both in high and low HDI regions. Relative frequencies of the 148 serovars were significantly affected only by HDI and volume. Considering that serovars representation can also be affected by water sample volume, efforts toward the standardization of water samplings for monitoring purposes should be considered. Further approaches such as metagenomics could provide more comprehensive insights about the microbial ecology of fresh water and its importance for the quality and safety of agricultural products.

Antimicrobial resistance in the globalized food chain: a One Health perspective applied to the poultry industry.

Antimicrobial resistance (AMR) remains a major global public health crisis. The food animal industry will face escalating challenges to increase productivity while minimizing AMR, since the global demand for animal protein has been continuously increasing and food animals play a key role in the global food supply, particularly broiler chickens. As chicken products are sources of low-cost, high-quality protein, poultry production is an important economic driver for livelihood and survival in developed and developing regions. The globalization of the food supply, markedly in the poultry industry, is aligned to the globalization of the whole modern society, with an unprecedented exchange of goods and services, and transit of human populations among regions and countries. Considering the increasing threat posed by AMR, human civilization is faced with a complex, multifaceted problem compromising its future. Actions to mitigate antimicrobial resistance are needed in all sectors of the society at the human, animal, and environmental levels. This review discusses the problems associated with antimicrobial resistance in the globalized food chain, using the poultry sector as a model. We cover critical aspects of the emergence and dissemination of antimicrobial resistance in the poultry industry and their implications to public health in a global perspective. Finally, we provide current insights using the multidisciplinary One Health approach to mitigate AMR at the human-animal-environment interface.

In ovo threonine supplementation affects ileal gene expression of nutrient transporters in broilers inoculated post-hatch with Salmonella Enteritidis.

The effect of in ovo threonine (Thr) supplementation on the ileal expression of glucose, peptide and amino acid transporters was assessed in Salmonella Enteritidis-challenged broiler chicks. At 17.5 days of incubation, fertile eggs were supplemented in the amniotic fluid with sterile saline or 3.5% threonine. Hatchlings were individually weighed, and Salmonella Enteritidis negative status was confirmed. At 2 days of age, half of the birds of each group were inoculated with sterile nutrient broth or Salmonella Enteritidis inoculum. Relative expression of sodium-dependent glucose transporter 1 (SGLT1), glucose transporter 2 (GLUT2), di- and tri-peptide transporter 1 (PepT1) and alanine, serine, cysteine, threonine transporter (ASCT1) was assessed at hatch, 2 and 9 days of age, i.e., before inoculation and 7 days post-inoculation (dpi). At 9 days of age (7dpi), threonine increased SGLT1 and GLUT2 expression, whereas GLUT2 expression decreased in Salmonella-challenged birds. There was a significant interaction between threonine and Salmonella for PepT1 and ASCT1. Threonine increased PepT1 expression only in non-challenged birds. In addition, in ovo supplementation increased expression of ASCT1 regardless of post-hatch inoculation; Salmonella inoculation resulted in decreased expression of ASCT1 only in supplemented birds. The results suggest that while intra-amniotic threonine administration in broiler embryos increases the expression of genes related to the absorption of monosaccharides and amino acids, Salmonella challenge may negatively affect the expression of protein related transporters in the ileum of broilers.

Staphylococcus sciuri as a Reservoir of mecA to Staphylococcus aureus in Non-Migratory Seabirds from a Remote Oceanic Island.

Aim: Genomic analysis of a methicillin-resistant Staphylococcus aureus (MRSA) strain cultured from a non-migratory seabird at Fernando de Noronha Archipelago (Brazilian oceanic islands) was carried out to investigate the potential origin of MRSA genetic determinants in an ecological setting with minimal or absent antimicrobial selective pressure, and minimal interaction with humans and domestic animals. Results: The study determined mecA gene homology and the phylogenetic relatedness with mecA described in Staphylococcus sciuri, which was the major Staphylococcus spp. cultured from the birds. Our findings corroborate in silico assumptions that the mecA gene in MRSA strains clinically relevant for humans and animals originates from S. sciuri ancestors. Conclusion: Coagulase-negative staphylococci seem to be natural reservoirs of methicillin-resistant genes to S. aureus, even in environments with very low antimicrobial selection pressure.

Occurrence of KPC-Producing Escherichia coli in Psittaciformes Rescued from Trafficking in Paraíba, Brazil.

The emergence and spread of antimicrobial resistance pose a threat to public health globally. Antibiotic-resistant bacteria and genes can disseminate among environments, animals and humans. Therefore, investigation into potential reservoirs of multidrug-resistant bacteria is of great importance to the understanding of putative transmission routes of resistant bacteria and resistance genes. This study aimed to report the occurrence of Escherichia coli harboring the Klebsiella pneumoniae carbapenemase-producing gene (blaKPC) in Psittaciformes rescued from wildlife trafficking in Paraíba State, Brazil. Cloacal swabs were collected from thirty birds and cultured by conventional microbiology using MacConkey and serum tryptone glucose glycerol (STGG) media supplemented with selective antimicrobials. E. coli isolates (n = 43) were identified by phenotypic tests and confirmed by MALDI-TOF. Antimicrobial susceptibility profiles were determined by means of Kirby-Bauer test. All isolates were further screened for extended-spectrum beta-lactamase (ESBL) production, and putative genes encoding ESBL were investigated by PCR. Additionally, blaKPC-harboring strains were genotyped by REP-PCR. A total of 43 E. coli phenotypically resistant isolates were recovered. The highest resistance rate was observed against ciprofloxacin. Among the resistance genes, only blaKPC was found in seven different birds from three species. According to the genotyping, these seven isolates belonged to four different strains. To date, this is the first report on the occurrence of KPC-E. coli in Psittaciformes rescued from trafficking in Northeastern Brazil. Due to the high clinical importance of KPC-E. coli, our findings suggest that wild animals in captivity at wildlife rescue centers can play a role as reservoirs of bacteria that are resistance to Critically Important antimicrobials in human medicine.

Expression of glucose transporters and morphometry in the intestine of Japanese quails after hatch.

The intestinal physiology and mechanisms involved in nutrient transport are not well established in quails (Coturnix coturnix japonica). The present study assessed the growth performance, morphological development, duodenal density and the expression of Sglt1 and Glut2 of female Japanese quails from 1 to 49 days of age. The three small intestine segments were sampled weekly from 1 to 49 days of age to evaluate villus height, crypt depth and villus: crypt ratio, and goblet cell counts. Scanning electronic microscopy was used to determine duodenal villus density, and real-time polymerase chain reaction (qPCR) was used to study the sodium/glucose cotransporter-1 Sglt1 and glucose transporter Glut2 in the jejunum. Villus height and crypt depth in the duodenum, jejunum and ileum increased with age until 42 and 49 days of age (P < 0.001), and regression analysis evidenced a quadratic effect (P < 0.0001), indicating increasing values to a maximum and then a decrease afterwards. Goblet cell counts increased (P < 0.001) in duodenum, jejunum and ileum from 1 to 42 days, decreasing at 49 days, which was also corroborated by the regression analysis. Villus density in the duodenum was greater in the first week, decreased with age and increased again at 42 days, probably due to the proximity with egg production onset. The expression of Sglt1 and Glut2 mRNA in the jejunum varied with age. In conclusion, the intestinal mucosa of female Japanese quail developed morphologically until 42days and functionally until earlier ages, indicating an adaptation to the exogenous diet during the first weeks of life.

High Incubation Temperature and Threonine Dietary Level Improve Ileum Response Against Post-Hatch Salmonella Enteritidis Inoculation in Broiler Chicks.

This study assessed the effect of both embryonic thermal manipulation and dietary threonine level on the response of broilers inoculated with Salmonella Enteritidis, considering bacterial counts in the cecal contents, intestinal morphology, mucin and heat shock protein 70 gene expression, body weight and weight gain. Thermal manipulation was used from 11 days of incubation until hatch, defining three treatments: standard (37.7°C), continuous high temperature (38.7°C) and continuous low temperature (36.7°C). After hatch, chicks were distributed according to a 3x2+1 factorial arrangement (three temperatures and two threonine levels and one sham-inoculated control). At two days of age, all chicks were inoculated with Salmonella Enteritidis, except for the sham-inoculated control group. There was no interaction between the factors on any analyses. High temperature during incubation was able to reduce colonization by Salmonella Enteritidis in the first days, reducing both Salmonella counts and the number of positive birds. It also increased mucin expression and decreased Hsp70 expression compared with other inoculated groups. High temperature during incubation and high threonine level act independently to reduce the negative effects associated to Salmonella Enteritidis infection on intestinal morphology and performance, with results similar to sham-inoculated birds. The findings open new perspectives for practical strategies towards the pre-harvest Salmonella control in the poultry industry.

Expression of heat shock protein in broiler embryo tissues after acute cold or heat stress.

This study evaluated the expression of heat shock protein 70 kD (hsp70) in broiler chicken embryos subjected to cold (Experiment I) or high incubation temperature (Experiment II). In each experiment, fertile eggs were distributed in three incubators kept at 37.8 degrees C. At day 13 (D13), D16, and D19 of incubation, the embryos were subjected to acute cold (32 degrees C) or heat (40 degrees C) for 4-6 hr. Immediately after cold or heat exposure, samples from the liver, heart, breast muscle, brain, and lungs of 40 embryos were taken per age and treatment (control or stressed embryos). A tissue pool from 10 embryos was used as 1 replication. The levels of hsp70 in each tissue sample was quantified by Western blot analysis. The data were analyzed in a 3 x 2 factorial arrangement of treatments with four replications. hsp70 was detected in all embryo tissues, and the brain contained 2- to 5-times more hsp70 protein compared to the other tissues in either cold or heat stressed embryos. hsp70 increases were observed in the heart and breast muscle of cold stressed embryos at D16 and D19, respectively. Heat stressed embryos showed an increase of hsp70 in the heart at D13 and D19, and in the lung at D19 of incubation. Younger embryos had higher hsp70 synthesis than older embryos, irrespective of the type of thermal stressor. The results indicate that the expression of hsp70 in broiler chicken embryos is affected by cold and heat distress, and is tissue- and age-dependent.