Fumonisin B (FB) and other fumonisins, deoxynivalenol (DON), and zearalenone (ZEN) are mycotoxins (secondary metabolites of fungi) present at high levels of contamination in poultry diets and threatening the sustainability of the poultry industry and egg safety for consumers. However, residual mycotoxins in breeder eggs and their effects on chicken progeny and gizzard ulcerations remain unclear. To unveil mycotoxin contaminations from daily diets to breeder eggs, 293 poultry feed samples were collected from three large-scale poultry provinces across Northern China to Southern China. Average levels of 1,628 ± 4.36 μg/kg of FB1, 593 ± 11.16 μg/kg of DON, 69 ± 9.21 μg/kg of ZEN, 52 ± 7.33 μg/kg of OTA, and 24 ± 5.85 μg/kg of AFB1 were found in feedstuffs and poultry diets using commercial ELISA kits. In terms of residual mycotoxins in breeder eggs, FB1 and DON contaminations dominated residues in egg albumen and yolk samples. Out of 221 breeder eggs, the average residual of FB1 in albumen were 320.6 ± 10.12 μg/kg (Hebei), 420.2 ± 10.98 μg/kg (Guangdong), and 549.4 ± 10.27 (Guangxi). Moreover, higher residual of DONs were determined in Guangdong and Guangxi provinces compared to Hebei province. ZEN, ochratoxins A (OTA), and aflatoxin B1 (AFB1) contamination at low levels were found in the above samples collected from afronmentioned three provinces. Based on residual mycotoxins in breeder eggs, SPF embryonated eggs aged 11 days were inoculated into albumen with different doses of FB1, FB2 or DON, or a combination of FB1 and DON, or a combination of FB1 with FB2 and FB3. A lower hatching rate was observed in the chicken progenies with the combination of 24 μg of FB1 and 0.1 μg of DON compared to other treatments. Moreover, typical gizzard ulcerations with hemorrhagic lungs were observed in the progeny of breeder eggs post-inoculation of 24 μg of FB1 and synergetic inoculation of FB1 and DON. Finally, residual FB mycotoxins were detected in the gizzards and in the lungs of the progenies. Based on the above evidence, feed-borne FB1 and DON are dominant mycotoxins in breeder eggs and threatening food security using breeder eggs as a Trojan horse. More importantly, the residual of FB1 alone and in combination with of DON contamination are associated with low hatching rate and gizzard ulcerations in chicken progenies, hampering sustainable development perspectives of the poultry industry.
Chlamydia psittaci (C. psittaci) is a common zoonotic agent that affects both poultry and humans. Interleukin 10 (IL-10) is an anti-inflammatory factor produced during chlamydial infection, while dendritic cells (DCs) are powerful antigen-presenting cells that induce a primary immune response in the host. However, IL-10 and DCs regulatory mechanisms in C. psittaci infection remain elusive. In vivo and in vitro investigations of the regulatory mechanisms were performed. IL-10−/− mice, conditional DCs depletion mice (zinc finger dendritic cell-diphtheria toxin receptor [zDC-DTR]), and double-deficient mice (DD, IL-10−/−/zDCDTR/DTR) were intranasally infected with C. psittaci. The results showed that more than 90% of IL-10−/− mice, 70% of wild-type mice, and 60% of double-deficient mice survived, whereas all zDC-DTR mice died. A higher lymphocyte proliferation index was found in the IL-10 inhibitor mice and IL-10−/− mice. Moreover, severe lesions and high bacterial loads were detected in the zDC-DTR mice compared with double-deficient mice. In vitro studies revealed increased OX40-OX40 ligand (OX40-OX40L) activation and CD4+T cell proliferation. Besides, the expression of indoleamine 2, 3-dioxygenase (IDO), and regulatory T cells were significantly reduced in the co-culture system of CD4+ T cells and IL-10−/− DCs in C. psittaci infection. Additionally, the activation of the NLR family pyrin domain-containing 3 (NLRP3) inflammasome increased to facilitate the apoptosis of DCs, leading to rapid clearance of C. psittaci. Our study showed that IL-10−/− upregulated the function of deficient DCs by activating OX40-OX40L, T cells, and the NLPR3 inflammasome, and inhibiting IDO, and regulatory T cells. These effects enhanced the survival rate of mice and C. psittaci clearance. Our research highlights the mechanism of IL-10 interaction with DCs, OX40-OX40L, and the NLPR3 inflammasome, as potential targets against C. psittaci infection.
Salpingitis is manifested as hemorrhagic follicular inflammation exudations and peritonitis, leading to reduced egg production and high culling of breeder flocks. From 2018 to 2021, increasing salpingitis during egg peak is threatening the poultry industry post-artificial insemination, both in breeder layers and breeder ducks across China. In our study, Escherichia coli (E. coli), Enterococcus faecalis (E. faecalis) and Chlamydia psittaci (C. psittaci) were isolated and identified from the diseased oviducts using biochemical tests and PCR. To identify and isolate pathogenicity, we inoculated the isolates into laying hens via an intravaginal route. Later, laying hens developed typical salpingitis after receiving the combination of the aforementioned three isolates (1 × 105 IFU/mL of C. psittaci and 1 × 106 CFU/mL of E. faecalis and E. coli, respectively), while less oviduct inflammation was observed in the layers inoculated with the above isolate alone. Furthermore, 56 breeder ducks were divided into seven groups, eight ducks per group. The birds received the combination of three isolates, synergic infection of E. coli and E. faecalis, and C. psittaci alone via vaginal tract, while the remaining ducks were inoculated with physiological saline as the control group. Egg production was monitored daily and lesions of oviducts and follicles were determined post-infection on day 6. Interestingly, typical salpingitis, degenerated follicles and yolk peritonitis were obviously found in the synergic infection of three isolates and the birds inoculated with C. psittaci alone developed hemorrhagic follicles and white exudates in oviducts, while birds with E. faecalis or E. coli alone did not develop typical salpingitis. Finally, higher E. coli loads were determined in the oviducts as compared to E. faecalis and C. psittaci infection. Taken together, the combination of E. coli and E. faecalis, and C. psittaci could induce typical salpingitis and yolk peritonitis both in laying hens and breeder ducks. Secondary infection of E. coli and E. faecalis via artificial insemination is urgently needed for investigation against salpingitis.
Background: Fumonisin B1 (FB1) is a secondary metabolite produced mainly by Fusarium verticillioides or Fusarium proliferatum. It poses a huge threat to the sustainable animal industry and human health as well via food chains (egg, meat and milk). Although E. coli-expressed nanobodies are documented for diagnostic applications, nanobodies remain elusive as FB1 detoxifiers in feed and food. Results: In the present study, the E. coli-expressed nanobody was assessed to remove FB1 in fresh milk, embryonated eggs and broilers. Firstly, 2 alpacas received intramuscularly FB1-adjuvanted BSA 6 times, and then the variable domain of the heavy-chain antibody (VHH) of fb1 genes were amplified to clone into the pCANTAB 5 E vector in order to generate a VHH-FB1 phage antibody display library, yielding 3.4 × 1010 capacity with 96.7% positivity. Afterwards, 5 anti-FB1 nanobodies were expressed and identified. Furthermore, maximal 43.2% FB1 was removed from milk by 1:2000 concentration of nanobody 5 (Nb5). Furthermore, SPF-embryonated eggs were inoculated into albumens with nanobody-treated FB1. The Nb5 group yielded an 83.3% hatching rate, higher body weight, lower gizzard ulceration and fewer FB1 residuals. In order to warrant the above results, 50 broilers aged 10 days were received orally with 20 ppm of FB1 for 20 days. At the same time, birds were fed orally with 50 μg of Nb5 or bivalent nanobody 11 (BiNb11). Finally, the Nb5 group showed a higher relative body weight gain and lower gastric ulcerations and fewer inflammations in the thymus and bursa. Conclusions: Based on the above evidence, the Nb5 nanobody may be considered as an additional FB1 detoxifier, contributing to FB1 decontamination.
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