Principais desafios que afetam a saúde intestinal das aves

O trato gastrointestinal das aves é colonizado por microrganismos benéficos e maléficos, vivendo de maneira comensal. As bactérias benéficas influenciam positivamente a integridade intestinal, funcionalidade da barreira de proteção, resposta imunológica, digestão e absorção dos nutrientes. As bactérias maléficas causadoras de enterites nas aves são: Clostridium colinum (enterite ulcerativa), Clostridium perfringens (enterite necrótica), Campylobacter jejuni e Campylobacter coli (enterite hemorrágica), Escherichia coli, Salmonella entérica (Salmonella pullorum - pulorose e Salmonella gallinarum - tifo aviário). Os protozoários causadores de enterites são: Eimeria acervulina, Eimeria maxima, Eimeria tenella (causam a coccidiose), Cryptosporidium meleagridis, Cryptosporidium baileyi e Cryptosporidium galli. Os vírus que acometem aves são: Paramoxivírus aviário tipo I (doença de Newcastle), rotavírus, astrovírus e reovírus. Porém, as lesões intestinais provocadas pelo microbioma maléfico podem ser amenizadas com o uso de prebióticos e probióticos na dieta das aves. Os prebióticos e probióticos fortalecem o sistema imunológico da ave, previne contra doenças entéricas infecciosas e melhora a digestão e absorção intestinal. Algumas bactérias que pertencem a microbiota intestinal das aves são utilizadas como probióticos, as espécies mais utilizadas pertencem ao gênero Lactobacillus e Bifidobacterium. Esta revisão analisa as informações científicas disponíveis sobre os principais microrganismos que afetam a saúde intestinal das aves e o uso de prebióticos e probióticos como aditivos preventivos na literatura vigente.

Anticoccidial resistance in poultry

This study aimed to determine the resistance of coccidia to ionophores used in broiler farms in Tizi-Ouzou province, Algeria. Droppings were collected and recovered Eimeria oocyst isolates were analysed by morphometry to determine their composition, and then inoculated by peros into chicks of the Arbor Acres strain, reared on the ground. Four of six groups of chicks were treated to test the sensitivity of oocysts to four anticoccidial agents added to their growth feeds [(robenidine (33 ppm), monensin (120 ppm), narasin-nicarbazin (80 ppm) and salinomycin (60 ppm)], while the other two groups were controls. The results revealed the presence of total resistance to monensin and robenidine, and partial resistance to salinomycin and the narasin-nicarbazin combination. The lack of sensitivity to monensin and robenidine was unsurprising, given their inappropriate and unreasonable use for years as the only anticoccidial compounds. The appearance of partial resistance to narasin-nicarbazin and salinomycin suggests the development of cross-resistance in the Eimeria population. The possibility of a relatively uniform composition of Eimeria species collected in these farms indicates that Eimeria acervulina and Eimeria maxima develop resistance more quickly to these ionophores. Finally, a control strategy must be rigorously developed by considering other molecules that are alternatives to anticoccidials.

Eimeria acervulina Microneme Protein 3 Inhibits Apoptosis of the Chicken Duodenal Epithelial Cell by Targeting the Casitas B-Lineage Lymphoma Protein

Eimeria acervulina (E. acervulina) causes coccidiosis in poultry which persists as economic pain worldwide. Most damage to the intestinal mucosa results from apoptosis of the infected intestinal epithelial cells. The Microneme protein 3 (MIC3) protein is a key virulence factor in some parasites involved in host cell apoptosis inhibition. Here, we studied whether and how MIC3 affects the apoptosis in E. acervulina infected chicken duodenal epithelial cells. Through flow cytometry (FCM), we found that the presence of merozoites and the overexpression of MIC3 significantly decreased apoptosis and the activity of caspase-3 in chicken duodenal epithelial cells at 4, 6, and 8 h post merozoite infection (P < 0.01). Silencing the Casitas B-lineage lymphoma (CBL) protein, a host receptor for MIC3 with shRNA was shown to promote apoptosis in the chicken duodenal epithelial cells. The early apoptotic rate of host cells in the lentiviral-MIC3 group was significantly lower than that in the lentiviral-MIC3 + shRNA CBL group at 4 h after MIC3 expression (P < 0.01), and it was moderately decreased in the lentiviral-MIC3 + shRNA CBL group compared with that in the shRNA CBL group. Our data indicated that MIC3 inhibited early apoptosis of E. acervulina infected chicken duodenal epithelial cells by targeting host receptor-CBL protein. These findings unveiled one of the mechanisms of how intracellular parasites affect the apoptosis of infected host cells, which provided a deeper understanding of their pathogenesis.

The impact of maturity on the ability of Eimeria acervulina and Eimeria meleagrimitis oocysts to sporulate

The sporulation of oocysts of Eimeria that infect poultry is known to be under the influence of environmental conditions, including temperature, oxygen supply, and moisture. However, even when these conditions are optimal, the level of sporulation can remain low. The effect of oocyst maturity on their ability to sporulate was investigated for two species of Eimeria: E. acervulina of chickens, and E. meleagrimitis of turkeys. After oral infection of birds, oocysts were collected at their production site in the intestine at different times around the prepatent period. The percentage of sporulation was determined by observation of 100 oocysts for each sample. With E. acervulina, it was observed that sporulation depended on the time of collection of the oocysts in the intestine, and that it increased with aging oocysts (from 5% to 40% globally in 8 h). With E. meleagrimitis, sporulation remained low with oocysts collected in the duodenum (below 20%), but oocysts collected in the midgut and in the lower intestine sporulated more efficiently (around 80%) than oocysts collected in the duodenum at the same time. One explanation for these results is the assumption that oocysts may be produced before fertilization, and that microgametes have not yet fertilized the newly produced oocysts. As time goes on, more oocysts would be fertilized, locally in the duodenum for E. acervulina, and descending along the gut for E. meleagrimitis. This hypothesis needs to be investigated further, but it could lead to new approaches to control these parasites by targeting the microgametes.

Glyceraldehyde-3-phosphate dehydrogenase from Eimeria acervulina modulates the functions of chicken dendritic cells to boost Th1 type immune response and stimulates autologous CD4+ T cells differentiation in-vitro

Dendritic cells (DCs) play a pivotal role to amplify antigen-specific immune responses. Antigens that sensitize T cells via antigen-presentation by DCs could enhance the capacity of host immunity to fight infections. In this study, we tested the immunogenic profiles of chicken DCs towards Glyceraldehyde-3-phosphate dehydrogenase from Eimeria acervulina (EaGAPDH). Immunoblot analysis showed that recombinant EaGAPDH (rEaGAPDH) protein was successfully recognized by rat sera generated against rEaGAPDH. Interaction and internalisation of rEaGAPDH by chicken splenic-derived DCs (chSPDCs) was confirmed by immunofluorescence analysis. Flow cytometry revealed that chSPDCs upregulated MHCII, CD1.1, CD11c, CD80, and CD86 cell-surface markers. Moreover, mRNA expressions of DC maturation biomarkers (CCL5, CCR7, and CD83) and TLR signalling genes (TLR15 and MyD88) were also upregulated whereas those of Wnt signalling were non-significant compared to negative controls. rEaGAPDH treatment induced IL-12 and IFN-γ secretion in chSPDCs but had no effect on IL-10 and TGF-β. Likewise, DC-T cell co-culture promoted IFN-γ secretion and the level of IL-4 was unaffected. Proliferation of T cells and their differentiation into CD3+/CD4+ T cells were triggered in chSPDCs-T cells co-culture system. Taken together, rEaGAPDH could promote Th1 polarization by activating both host DCs and T cells and sheds new light on the role of this important molecule which might contribute to the development of new DCs-based immunotherapeutic strategies against coccidiosis.

Isolated and combined organic acids in diets of broiler chickens challenged with Eimeria acervulina

ABSTRACT This study aimed to evaluate the effect of isolated or combined citric and benzoic acids added to the diets of broiler chickens on performance, allometry of the digestive system organs, intestinal pH and quantity of microorganisms in the jejunum. A total of 840 male Cobb broiler chicks were utilized, distributed in a complete random design in 2 × 2 factorial scheme, supplemented or not with citric acid, and sodium benzoate, with seven replications. At 14 days old, 1mL of a solution containing 1 × 105 sporulated oocysts of Eimeria acervulina per bird was inoculated orally. There was no effect of the acids on the broiler’s performance in the 1 to 21-day period. In the total period (1 to 42 days), the broilers fed with a blend of citric and benzoic acid presented greater feed intake, without increment in weight gain. The data of allometry of the digestive system organs and the jejunal pH values were not influenced by the treatments. The citric acid increased the bacterial quantity of gram-positive coccus and total anaerobes in the jejunum. Under the conditions of the present study, we conclude that the citric and benzoic acids, isolated or combined, do not benefit the nutrition of broilers challenged with E. acervulina.