scholarly journals Recombinant HcGAPDH Protein Expressed on Probiotic Bacillus subtilis Spores Protects Sheep from Haemonchus contortus Infection by Inducing both Humoral and Cell-mediated Responses

2019 ◽  
Author(s):  
Yi Yang ◽  
Guiheng Zhang ◽  
Jie Wu ◽  
Xueqiu Chen ◽  
Danni Tong ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Immune Responses ◽  
Haemonchus Contortus ◽  
Mononuclear Cells ◽  
Parasitic Infection ◽  
Small Ruminants ◽  
Infected Sheep ◽  
Infection Model ◽  
Average Weight ◽  
Gastrointestinal Microbiota

AbstractProbiotic Bacillales have been shown effective in controlling pathogens. In particular, live probiotic bacteria are thought to improve the composition of gastrointestinal microbiota, and to reduce pathogen colonization. However, how probiotics regulate immune responses and protect the host from parasitic infection remains largely unknown. In this study, we investigated whether Bacillales can be used against Haemonchus contortus, a parasitic nematode that infects small ruminants in sheep and goats worldwide. Using 16S ribosomal RNA sequencing, we found that Bacillales was highly depleted in the abomasal microbiota of sheep infected with H. contortus. We constructed a recombinant Bacillus subtilis strain (rBSCotB-HcG) that express glyceraldehyde-3-phosphate dehydrogenase of H. contortus (HcGAPDH) on its spore surface. However, mice orally administrated with the rBSCotB-HcG strain showed strong Th1-dominated immune responses; and sheep administrated per os with rBSCotB-HcG showed increased proliferation of peripheral blood mononuclear cells, elevated anti-HcGAPDH IgG levels in sera, and higher anti-HcGAPDH sIgA levels in intestinal mucus. In addition, treatment of H. contortus infected sheep with rBSCotB-HcG (Hc+rBSCotB-HcG) promoted the abundance of probiotic species in the abomasal microbiota; it also improved the average weight gain of the sheep by 27.7%. These Hc+rBSCotB-HcG sheep have reduced number of eggs per gram of feces (by 84.1%) and worm burdens (by 71.5%), with alleviated abomasal damage by H. contortus. Collectively, our data demonstrate the protective roles of CotB-HcGAPDH-expressing B. subtilis spores against H. contortus infection, suggesting a potential value of using this probiotic-based strategy in controlling parasitic nematodes of socioeconomic importance.ImportanceSequencing of the infected sheep ’s stomach flora revealed potential probiotics that could control H. contortus infection, and further genetically engineered recombinant probiotic spores expressing parasite protein, and validated their good immunogenicity in a mouse model. In the sheep infection model, the recombinant probiotics have proven to be effective against parasite infections.

scholarly journals Recombinant HcGAPDH Protein Expressed on Probiotic Bacillus subtilis Spores Protects Sheep from Haemonchus contortus Infection by Inducing both Humoral and Cell-Mediated Responses

mSystems ◽  
2020 ◽  
Vol 5 (3) ◽  
Author(s):  
Yi Yang ◽  
Guiheng Zhang ◽  
Jie Wu ◽  
Xueqiu Chen ◽  
Danni Tong ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Weight Gain ◽  
16S Rrna ◽  
Immune Responses ◽  
Haemonchus Contortus ◽  
Infected Sheep ◽  
16S Rrna Sequencing ◽  
Probiotic Bacteria ◽  
Content Type ◽  
Rrna Sequencing

ABSTRACT Probiotic Bacillales are effective in controlling pathogens. Live probiotic bacteria improve the composition of the gastrointestinal microbiota, leading to a reduction in pathogen colonization. However, it remains largely unknown how probiotics regulate the host’s immunologic responses and protect the host from parasitic infection. In this study, we addressed whether Bacillales were effective against Haemonchus contortus, a parasitic nematode that infects small ruminants worldwide. Using 16S rRNA sequencing, we found that Bacillales were largely depleted in the abomasal microbiota of sheep infected with H. contortus. We constructed a recombinant Bacillus subtilis named rBSCotB-HcG that express the glyceraldehyde-3-phosphate dehydrogenase of H. contortus (HcGAPDH) on its spore surface using the Bacillus subtilis spore coat protein B (CotB) as a carrier. Mice receiving rBSCotB-HcG orally showed strong Th1-dominated immune responses. More importantly, sheep administered BSCotB-HcG per os showed increasing proliferation of the peripheral blood mononucleates, elevated anti-HcGAPDH IgG in sera, and higher anti-HcGAPDH sIgA in the intestinal mucus than the control sheep. The average weight gain of H. contortus-infected sheep treated with rBSCotB-HcG (Hc+rBSCotB-HcG) was 48.73% greater than that of unvaccinated sheep. Furthermore, these Hc+rBSCotB-HcG sheep had fewer eggs per gram of feces by 84.1% and adult worms by 71.5%. They also demonstrated greatly lessened abomasal damage by H. contortus with an abundance of probiotic species in the abomasal microbiota. Collectively, our data unequivocally demonstrate the protective roles of CotB-HcGAPDH-expressing B. subtilis spores in against H. contortus infection and showed great potential of using probiotic-based strategy in controlling parasitic nematodes of socioeconomic importance in general. IMPORTANCE Initial analyses of the abomasal microbiota of sheep using 16S rRNA sequencing suggested that probiotic bacteria played a protective role in against H. contortus infection. A recombinant Bacillus subtilis expressing a fusion protein CotB-HcGAPDH on its spore’s surface induced strong Th1 immune response in a murine model. The same probiotic recombinant, upon only one oral application, protected sheep against H. contortus infection by reducing egg shedding and decreasing adult worm loads of the parasite and increasing body weight gain of infected sheep. Both Th1 and Th2 immune responses were evident in these immunized sheep.

scholarly journals Characterization of a Rhodanese Homologue from Haemonchus Contortus and its Immune-Modulatory Effects on Goat Immune Cells in Vitro

2020 ◽  
Author(s):  
Yujian Wang ◽  
Muhammad Ehsan ◽  
Jianmei Huang ◽  
Kalibixiati Aimulajiang ◽  
RuoFeng Yan ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Mononuclear Cells ◽  
Small Ruminants ◽  
Parasitic Nematodes ◽  
Dependent Manner ◽  
Peripheral Blood Mononuclear ◽  
Nematode Parasites ◽  
Wide Range

Abstract Background: Suppression and modulation of the immune response of the host by nematode parasites have been reported widely. Rhodaneses or thiosulfate: cyanide sulfurtransferases are present in a wide range of organisms, such as archea, bacteria, fungi, plants and animals. Previously, it was reported that a rhodanese homology could bind by goat peripheral blood mononuclear cells (PBMCs) in vivo.Results: In the present study, we cloned and produced recombinant rhodanese protein originated from Haemonchus contortus (rHCRD), which was one of the parasitic nematodes of small ruminants. The effect of this protein on modulating the immunity of goat PBMC and monocyte was studied in the current work. The predominant localization of the natural HCRD protein was verified as the bowel wall and body surface of worms, according to the immunohistochemical tests. It was proved in this study that the serum produced by artificially infecting goats with H. contortus successfully recognized rHCRD which conjugated goat PBMCs. The rHCRD was co-incubated with goat PBMCs to observe the immunomodulatory effect on proliferation, apoptosis and secretion of cytokines exerted by HCRD. The results showed that the interaction of rHCRD suppressed proliferation of goat PBMCs stimulated by ConA but did not induce the apoptosis of goat PBMCs. After rHCRD exposure, the production of TNF-α and IFN-γ were significantly decreased, however, it significantly increased the secretion of IL-10 and TGF-β1 in goat PBMCs. Phagocytotic assay by FITC-dextran internalization showed that rHCRD inhibited the phagocytosis of goat monocytes. Moreover, rHCRD could down-regulate the expression of MHC-II on goat monocytes in a dose-dependent manner. Conclusions: These discoveries proposed a possible target as immunomodulator, which was potentially beneficial to illuminate the interaction between parasites and hosts in the molecular level and hunt for innovative protein species as candidate targets of drug and vaccine.

scholarly journals Characterization of a rhodanese homologue from Haemonchus contortus and its immune-modulatory effects on goat immune cells in vitro

2020 ◽  
Author(s):  
Yujian Wang ◽  
Muhammad Ehsan ◽  
Jianmei Huang ◽  
Kalibixiati Aimulajiang ◽  
RuoFeng Yan ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Mononuclear Cells ◽  
Small Ruminants ◽  
Dependent Manner ◽  
Peripheral Blood Mononuclear ◽  
Mhc I ◽  
Nematode Parasites ◽  
Wide Range

Abstract Background: Modulation of the host immune response by nematode parasites has been widely reported. Rhodaneses (thiosulfate: cyanide sulfurtransferases) are present in a wide range of organisms, such as archaea, bacteria, fungi, plants and animals. Previously, it was reported that a rhodanese homologue could be bound by goat peripheral blood mononuclear cells (PBMCs) in vivo.Methods: In the present study, we cloned and produced a recombinant rhodanese protein originating from Haemonchus contortus (rHCRD), a parasitic nematode of small ruminants. rHCRD was co-incubated with goat PBMCs to assess its immunomodulatory effects on proliferation, apoptosis and cytokine secretion.Results: We verified that the natural HCRD protein localized predominantly to the bowel wall and body surface of the parasite. We further demonstrated that serum produced by goats artificially infected with H. contortus successfully recognized rHCRD, which bound to goat PBMCs. rHCRD suppressed proliferation of goat PBMCs stimulated by concanavalin A but did not induce apoptosis in goat PBMCs. The production of TNF-α and IFN-γ decreased significantly, whereas secretion of IL-10 and TGF-β1 increased, in goat PBMCs after exposure to rHCRD. rHCRD also inhibited phagocytosis by goat monocytes. Moreover, rHCRD downregulated the expression of major histocompatibility complex (MHC)-II on goat monocytes in a dose-dependent manner, but did not alter MHC-I expression.Conclusions: These results propose a possible immunomodulatory target that may help illuminate the interactions between parasites and their hosts at the molecular level and reveal innovative protein species as candidate drug and vaccine targets.

scholarly journals Characterization of a rhodanese homologue from Haemonchus contortus and its immune-modulatory effects on goat immune cells in vitro

2020 ◽  
Author(s):  
Yujian Wang ◽  
Muhammad Ehsan ◽  
Jianmei Huang ◽  
Kalibixiati Aimulajiang ◽  
RuoFeng Yan ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Mononuclear Cells ◽  
Small Ruminants ◽  
Dependent Manner ◽  
Peripheral Blood Mononuclear ◽  
Mhc I ◽  
Nematode Parasites ◽  
Wide Range

Abstract Background: Modulation of the host immune response by nematode parasites has been widely reported. Rhodaneses (thiosulfate: cyanide sulfurtransferases) are present in a wide range of organisms, such as archaea, bacteria, fungi, plants and animals. Previously, it was reported that a rhodanese homologue could be bound by goat peripheral blood mononuclear cells (PBMCs) in vivo.Methods: In the present study, we cloned and produced a recombinant rhodanese protein originating from Haemonchus contortus (rHCRD), a parasitic nematode of small ruminants. rHCRD was co-incubated with goat PBMCs to assess its immunomodulatory effects on proliferation, apoptosis and cytokine secretion.Results: We verified that the natural HCRD protein localized predominantly to the bowel wall and body surface of the parasite. We further demonstrated that serum produced by goats artificially infected with H. contortus successfully recognized rHCRD, which bound to goat PBMCs. rHCRD suppressed proliferation of goat PBMCs stimulated by concanavalin A but did not induce apoptosis in goat PBMCs. The production of TNF-α and IFN-γ decreased significantly, whereas secretion of IL-10 and TGF-β1 increased, in goat PBMCs after exposure to rHCRD. rHCRD also inhibited phagocytosis by goat monocytes. Moreover, rHCRD down-regulated the expression of major histocompatibility complex (MHC)-II on goat monocytes in a dose-dependent manner, but did not alter MHC-I expression. Conclusions: These results propose a possible immunomodulatory target that may help illuminate the interactions between parasites and their hosts at the molecular level and reveal innovative protein species as candidate drug and vaccine targets.

scholarly journals Proteomic identification of galectin-11 and 14 ligands fromHaemonchus contortus

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e4510
Author(s):  
Dhanasekaran Sakthivel ◽  
Jaclyn Swan ◽  
Sarah Preston ◽  
MD Shakif-Azam ◽  
Pierre Faou ◽  
...  
Keyword(s):  
Immune Responses ◽  
Adult Stage ◽  
Parasitic Infection ◽  
Small Ruminants ◽  
Affinity Column ◽  
Protein Targets ◽  
Zinc Metalloprotease ◽  
Host Parasite Interactions ◽  
Affinity Column Chromatography ◽  
Host Parasite

Haemonchus contortusis the most pathogenic nematode of small ruminants. Infection in sheep and goats results in anaemia that decreases animal productivity and can ultimately cause death. The involvement of ruminant-specific galectin-11 (LGALS-11) and galectin-14 (LGALS-14) has been postulated to play important roles in protective immune responses against parasitic infection; however, their ligands are unknown. In the current study, LGALS-11 and LGALS-14 ligands inH. contortuswere identified from larval (L4) and adult parasitic stages extracts using immobilised LGALS-11 and LGALS-14 affinity column chromatography and mass spectrometry. Both LGALS-11 and LGALS-14 bound more putative protein targets in the adult stage ofH. contortus(43 proteins) when compared to the larval stage (two proteins). Of the 43 proteins identified in the adult stage, 34 and 35 proteins were bound by LGALS-11 and LGALS-14, respectively, with 26 proteins binding to both galectins. Interestingly, hematophagous stage-specific sperm-coating protein and zinc metalloprotease (M13), which are known vaccine candidates, were identified as putative ligands of both LGALS-11 and LGALS-14. The identification of glycoproteins ofH. contortusby LGALS-11 and LGALS-14 provide new insights into host-parasite interactions and the potential for developing new interventions.

scholarly journals Proteomic identification of Galectin-11 and 14 ligands from Haemonchus contortus

2017 ◽  
Author(s):  
Dhanasekaran Sakthivel ◽  
Jaclyn Swan ◽  
Sarah Preston ◽  
MD Shakif-Azam ◽  
Pierre Faou ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Adult Stage ◽  
Parasitic Infection ◽  
Small Ruminants ◽  
Affinity Column ◽  
Protein Targets ◽  
Zinc Metalloprotease ◽  
Host Parasite Interactions ◽  
Affinity Column Chromatography ◽  
Host Parasite

Haemonchus contortus is the most pathogenic nematode of small ruminants. Infection in sheep and goats results in anaemia that decreases animal productivity and can ultimately cause death. The involvement of ruminant-specific galectin-11 (LGALS-11) and galectin-14 (LGALS-14) has been postulated to play important roles in protective immune responses against parasitic infection; however, their ligands are unknown. In the current study, LGALS-11 and LGALS-14 ligands in H. contortus were identified from larval (L4) and adult parasitic stages extracts using immobilised LGALS-11 and LGALS-14 affinity column chromatography and mass spectrometry. Both LGALS-11 and LGALS-14 bound more putative protein targets in the adult stage of H. contortus (43 proteins) when compared to the larval stage (2 proteins). Of the 43 proteins identified in the adult stage, 34 and 35 proteins were bound by LGALS-11 and LGALS-14, respectively, with 26 proteins binding to both galectins. Interestingly, hematophagous stage-specific sperm-coating protein and zinc metalloprotease (M13), which are known vaccine candidates, were identified as putative ligands of both LGALS-11 and LGALS-14. The identification of glycoproteins of H. contortus by LGALS-11 and LGALS-14 provide new insights into host-parasite interactions and the potential for developing new interventions.

scholarly journals Prevalence of gastrointestinal parasite in small ruminants of District Dir Upper Khyber Pakhtunkhwa Province of Pakistan

2023 ◽  
Vol 83 ◽  
Author(s):  
Ruhoollah ◽  
W. Khan ◽  
O. A. Al-Jabr ◽  
T. Khan ◽  
A. Khan ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Fasciola Hepatica ◽  
Intestinal Parasites ◽  
Parasitic Infection ◽  
Small Ruminants ◽  
Cost Effective ◽  
Gastrointestinal Parasites ◽  
Khyber Pakhtunkhwa ◽  
Khyber Pakhtunkhwa Province ◽  
Fasciola Spp

Abstract The present research was planned to assess the occurrence of intestinal parasites in small ruminants of Upper Dir of Khyber Pakhtunkhwa Province of Pakistan. For this purpose, the faecal material was collected randomly with gloved fingers directly from the rectum region of sheep and goats and the faecal materials were then put in hygienic plastic bottles with 10% formalin. The overall 315 (n=184 sheep and n= 131 goats) faecal samples were collected out of 315 samples, 281 were found positive for different parasites. Patterns-wise prevalence of GI parasites of the study area was found. Overall Single parasitism 89.20% (281/315) with 94.0% (173/184) in sheep and 82.43% (108/131) in goats. Double parasitic infection in small ruminant recorded in which Fasciola+ Haemonchus. contortus in sheep were found their prevalence was 25.54% (47/184). While in goats, the double parasitic infection in which Haemonchus contortus+Trichuris spp were found and their prevalence were 23.43% (30/131). The species found in the sample of sheep were includes, i.e., Strongyloides papillosus (41.30%), Heamonchus controtus (21.73%), Trichuris ovis (17.39%), and Fasciola hepatica (13.58%), the corresponding value for goat were Strongyloides spp 33.33% (36/108), Haemonchus spp 28.70%, (27/108), Trichuris spp 25.20% (27/184) and Fasciola spp 10.68% (14/184). The sheep of the study area are more infected as compared to goats. This study suggested that gastrointestinal parasites are major health problems of small ruminants in the study area. Therefore, a comprehensive study on species of gastrointestinal parasites circulating in the area, control options, cost-effective strategies and awareness about gastrointestinal parasites among the farmers in the study area should be instituted.

scholarly journals Etiopathological and hematobiochemical profiles in goats with gastrointestinal disorders

2021 ◽  
pp. 1760-1766
Author(s):  
Sunena Nayak ◽  
Prasana Kumar Rath ◽  
Susen Kumar Panda ◽  
Bidyut Prava Mishra ◽  
Rajshree Mishra ◽  
...  
Keyword(s):  
Risk Factors ◽  
Production Efficiency ◽  
Management Practices ◽  
Mononuclear Cells ◽  
Parasitic Infection ◽  
Clinical Signs ◽  
Small Ruminants ◽  
Blood Analysis ◽  
Parasitic Infections ◽  
Chi Square Analysis

Background and Aim: Gastrointestinal (GI) disorders in small ruminants limit production efficiency and productivity growth in the livestock sector, thereby directly preventing farmers from augmenting their income. This study aimed to provide detailed insight into the etiology, hematobiochemical parameters, and epidemiological risk factors of GI disorders in goats and to determine the pathology associated with the disorders. Materials and Methods: Over the period of 2018-2019, 500 goats in and around Bhubaneswar, Odisha, India, were screened for GI disorders based on clinical signs. Blood samples from the control (n=10) and treatment (n=25) groups were collected for both hematological and serum biochemical alterations. Fecal examinations (n=220) were conducted for parasitic, bacterial, and virological assessments. Detailed necropsy and histopathological evaluations were conducted on 27 goats. Results: The GI disorder prevalence rate and mortality rate among the 500 goats analyzed were 44.4% and 12.27%, respectively. Chi-square analysis showed a significantly higher occurrence of GI disorders among the goats that were between 6 months and 1.5 years old (58.72%), were of the Ganjam breed (45.49%), had a poor body condition (71.11%), and were housed with an earthen floor (55.22%). The most common etiological risk factor observed was parasitic infection (65.45%), followed by bacterial (18.18%) and mixed infection (9.54%). Blood analysis showed neutrophilia and eosinophilia in infected goats, in addition to anemia; significant decreases in total protein, globulin, albumin, and glucose levels; and significant increases in aspartate transaminase and alanine aminotransferase levels. The major histopathological findings were infiltration of mononuclear cells and desquamation of the intestinal and ruminal mucosa. Conclusion: Stakeholders should focus not only on parasitic infections and other important etiological risk factors for GI disorders in goats but also on proper farming management practices to help enhance the income of farmers. The hematobiochemical alterations and pathomorphological changes reported in this study can be used by field veterinarians as guidelines for clinical evaluation and disease severity assessment.

scholarly journals Characterization of a rhodanese homologue from Haemonchus contortus and its immune-modulatory effects on goat immune cells in vitro

2020 ◽  
Author(s):  
Yujian Wang ◽  
Muhammad Ehsan ◽  
Jianmei Huang ◽  
Kalibixiati Aimulajiang ◽  
RuoFeng Yan ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Mononuclear Cells ◽  
Small Ruminants ◽  
Dependent Manner ◽  
Peripheral Blood Mononuclear ◽  
Mhc I ◽  
Nematode Parasites ◽  
Candidate Drug ◽  
Wide Range

Abstract Background: Modulation of the host immune response by nematode parasites has been widely reported. Rhodaneses (thiosulfate: cyanide sulfurtransferases) are present in a wide range of organisms, such as archaea, bacteria, fungi, plants and animals. Previously, it was reported that a rhodanese homologue could be bound by goat peripheral blood mononuclear cells (PBMCs) in vivo.Methods: In the present study, we cloned and produced a recombinant rhodanese protein originating from Haemonchus contortus (rHCRD), a parasitic nematode of small ruminants. rHCRD was co-incubated with goat PBMCs to assess its immunomodulatory effects on proliferation, apoptosis and cytokine secretion.Results: We verified that the natural HCRD protein localized predominantly to the bowel wall and body surface of the parasite. We further demonstrated that serum produced by goats artificially infected with H. contortus successfully recognized rHCRD, which bound to goat PBMCs. rHCRD suppressed proliferation of goat PBMCs stimulated by concanavalin A but did not induce apoptosis in goat PBMCs. The production of TNF-α and IFN-γ decreased significantly, whereas secretion of IL-10 and TGF-β1 increased, in goat PBMCs after exposure to rHCRD. rHCRD also inhibited phagocytosis by goat monocytes. Moreover, rHCRD down-regulated the expression of major histocompatibility complex (MHC)-II on goat monocytes in a dose-dependent manner, but did not alter MHC-I expression.Conclusions: These results propose a possible immunomodulatory target that may help illuminate the interactions between parasites and their hosts at the molecular level and reveal innovative protein species as candidate drug and vaccine targets.

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