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This experiment was motivated by the need to understand the impacts of delaying the first colostrum feeding on the prevalence of failed transfer of passive immunity (FTPI). A cohort of 216 kids was stratified into groups based on the colostrum feeding delay postbirth: 0-4 h, 4-8 h, 8-12 h, and 12-16 h. All kids received a single colostrum meal of 300 mL, and blood samples were collected approximately 36 h after feeding. Serum immunoglobulin G (SIgG) was measured using ELISA, and serum total protein (STP) was assessed using the Bradford method and refractometry (STP b and STP r ). Statistical methods like Pearson correlations, Bland-Altman plots, and Lin's concordance coefficient were employed to assess associations and agreements between SIgG, STP b and STP r . Receiver operator characteristic analysis was employed to determine optimal STP b and STP r thresholds for predicting FTPI (SIgG < 12 g/L). Subsequently, areas under the curve, sensitivity, and specificity were examined to assess the accuracy of these thresholds. Our results showed that for each hour's delay from birth to colostrum intake (up to 16 h), IgG apparent efficiency of absorption (AEA) decreases at an approximate rate of 2.0% per hour, and SIgG decreases at an approximate rate of 1.0 g/L per hour. However, this decline is not constant over time but intensifies progressively with increased feeding delay. Specifically, reductions in IgG AEA were 1.3, 2.9, and 5.9% per hour, and decreases in SIgG were 0.2, 0.3, and 0.7 g per hour for SIgG across the time intervals of 0-4 to 4-8 h, 4-8 to 8-12 h, and 8-12 to 12-16 h, respectively. Additionally, there was an increase in SIgG of 1.2 g/dL but a decrease in IgG AEA of 1.9% for each gram per kg of BW increase in IgG intake. The correlations between SIgG and STP r and STP b were 0.62, and 0.36, respectively. Optimal STP r and STP b thresholds predicting FTPI were determined to be 4.6 and 6.2 g/dL. The prevalence of FTPI, according to SIgG, STP r , and STP b thresholds were 63, 62, and 45%. Overall, STP r showed higher values for key performance metrics (i.e., sensitivity, likelihood ratio of positive tests, overall accuracy, and Youden's index), indicating better prediction ability than STP b . Our findings corroborate the critical importance of swift colostrum administration, ideally occurring no later than 12 h postbirth. Moreover, our research validates the effectiveness of Brix refractometry as a practical, on-farm method for assessing FTPI in goat kids., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Mastitis, an inflammation of the mammary gland affecting milk production and quality in dairy herds, is often associated with Staphylococcus spp. in goats. Neutrophils are crucial in combating infections by migrating into milk and deploying various defense strategies, including the release of neutrophil extracellular traps (NETs) composed of DNA, histones, and bactericidal proteins. This study investigated whether NETs are released by goat neutrophils stimulated in vitro by Staphylococcus aureus and Staphylococcus warneri, two common pathogens of goat mastitis. PMNs were isolated from blood from healthy adult goats. We evaluated goat NET formation by stimulating cells with: phorbol 12-myristate 13-acetate (PMA) as a positive control, cytochalasin for inhibition of actin polymerization, S. aureus, and S. warneri. NET formation was observed in response to chemical stimulation and bacterial presence, effectively trapping pathogens. Variations in NET formation between S. aureus and S. warneri suggest pathogen-specific responses. These findings suggest that the formation of NETs may be an important complementary mechanism in the defense against mastitis in goats. In conclusion, this study unveils a novel defense mechanism in goats, indicating the role of NETs against S. aureus and S. warneri in mastitis., Competing Interests: Declaration of Competing Interest none, (Copyright © 2024 Elsevier B.V. All rights reserved.)

Hybridization can result in the transfer of adaptive genetic material from one species to another, known as adaptive introgression. Bottlenecked (and hence genetically depleted) species are expected to be particularly receptive to adaptive introgression, since introgression can introduce new or previously lost adaptive genetic variation. The Alpine ibex (Capra ibex), which recently recovered from near extinction, is known to hybridize with the domestic goat (Capra aegagrus hircus), and signals of introgression previously found at the major histocompatibility complex were suggested to potentially be adaptive. Here, we combine two ancient whole genomes of Alpine ibex with 29 modern Alpine ibex genomes and 31 genomes representing six related Capra species to investigate the genome-wide patterns of introgression and confirm the potential relevance of immune loci. We identified low rates of admixture in modern Alpine ibex through various F statistics and screening for putative introgressed tracts. Further results based on demographic modelling were consistent with introgression to have occurred during the last 300 years, coinciding with the known species bottleneck, and that in each generation, 1-2 out of 100 Alpine ibex had a domestic goat parent. The putatively introgressed haplotypes were enriched at immune-related genes, where the adaptive value of alternative alleles may give individuals with otherwise depleted genetic diversity a selective advantage. While interbreeding with domestic species is a prevalent issue in species conservation, in this specific case, it resulted in putative adaptive introgression. Our findings highlight the complex interplay between hybridization, adaptive evolution, and the potential risks and benefits associated with anthropogenic influences on wild species., (© 2024 The Author(s). Molecular Ecology published by John Wiley & Sons Ltd.)

This study aimed to clarify the composition and bioactivity differences between goat and cow milk fat globule membrane (MFGM) protein by proteomic, and the immunomodulatory activity of MFGM proteins was further evaluated by using mouse splenic lymphocytes in vitro. A total of 257 MFGM proteins showed significant differences between goat and cow milk. The upregulated and unique MFGM proteins in goat milk were significantly enriched in the positive regulation of immune response, negative regulation of Interleukin-5 (IL-5) secretion, and involved in nucleotide-binding oligomerization domain (NOD)-like receptor signaling. The contents of IL-2 and Interferon-γ in the supernatant of spleen lymphocytes treated with goat MFGM proteins were much higher than those of IL-4 and IL-5, suggesting a Th1-skewed immune response. These results revealed that goat MFGM proteins could possess better immunomodulatory effects as compared to cow milk. Our findings may provide new insights to elucidate the physiological functions and nutritional of goat milk., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Milk is a whitish liquid that is secreted from mammary glands; and considered as the primary source of nutrition for newborns since they are not able to digest solid food. However, it contains primary nutrients, as well as growth and immune factors. Early weaning is a critical issue that face women and their babies in developing countries. To avoid infant malnutrition, they tend to use other milk types instead of baby formula. Therefore, the present study aimed to evaluate the impact of cow, buffalo, goat or camel milk consumption on oxidative stress, inflammation and immune response in male and female Sprague Dawley rats post weaning time. The amino acids, fatty acids, minerals and vitamins in the tested milk types were evaluated. Animals were divided into 5 groups (control, cow, buffalo, goat and camel milk administrated groups) (10 rats/group); each animal was administrated by 3.4 ml/day. Rats were administered with milk for 6 weeks; at the end of the 5th week, five animals of each group were isolated and the remaining five animals were immunized with sheep red blood cells (SRBCs) and kept for another week to mount immune response. The effect of different milk types on rats' immune response towards SRBCs was evaluated through pro-inflammatory cytokines, antioxidants, ESR and CRP measurement; together, with the histopathological examination of spleen samples and hemagglutination assay. Camel milk consumption reduced oxidative stress and inflammation in spleen that resulted from SRBCs immunization; in addition to, B cell stimulation that was apparent from the high level of anti-SRBCs antibodies. Camel milk is recommended for newborn consumption, due to its high-water content, unsaturated fatty acids, and vitamin C, as well as low lactose and fat content., (© 2024. The Author(s).)

Peste des petits ruminants is an acute and highly contagious disease caused by the Peste des petits ruminants virus (PPRV). Host proteins play a crucial role in viral replication. However, the effect of fusion (F) protein-interacting partners on PPRV infection is poorly understood. In this study, we found that the expression of goat plasminogen activator urokinase (PLAU) gradually decreased in a time- and dose-dependent manner in PPRV-infected goat alveolar macrophages (GAMs). Goat PLAU was subsequently identified using co-immunoprecipitation and confocal microscopy as an F protein binding partner. The overexpression of goat PLAU inhibited PPRV growth and replication, whereas silencing goat PLAU promoted viral growth and replication. Additionally, we confirmed that goat PLAU interacted with a virus-induced signaling adapter (VISA) to antagonize F-mediated VISA degradation, increasing the production of type I interferon. We also found that goat PLAU reduced the inhibition of PPRV replication in VISA-knockdown GAMs. Our results show that the host protein PLAU inhibits the growth and replication of PPRV by VISA-triggering RIG-I-like receptors and provides insight into the host protein that antagonizes PPRV immunosuppression.IMPORTANCEThe role of host proteins that interact with Peste des petits ruminants virus (PPRV) fusion (F) protein in PPRV replication is poorly understood. This study confirmed that goat plasminogen activator urokinase (PLAU) interacts with the PPRV F protein. We further discovered that goat PLAU inhibited PPRV replication by enhancing virus-induced signaling adapter (VISA) expression and reducing the ability of the F protein to degrade VISA. These findings offer insights into host resistance to viral invasion and suggest new strategies and directions for developing PPR vaccines., Competing Interests: The authors declare no conflict of interest.

In dairy operations, antibiotics have traditionally been used to treat, prevent, and control diseases. However, given the mounting global crisis of antimicrobial resistance (AMR), farmers are urged to re-assess and reduce their reliance on antibiotics. Thus, this randomized, double-blinded cohort study aimed to estimate the prevalence of failed and successful transfer of passive immunity (FTPI and STPI) in dairy goat kids reared under commercial conditions, and the effects of antibiotic metaphylaxis on the pre-weaning (≤42 d old) mortality in FTPI and STPI kids. Plasma concentration of immunoglobulin G at 1d old (pIgG-24 h) was measured in 747 male Saanen kids for the determination of FTPI and STPI (pIgG-24 h < 12 and ≥12 g/L, respectively). Kids were then randomly divided into two groups: those receiving a single penicillin injection at 1 d old (PEN), and those receiving no treatment (CTR). The mean (±SD) pIgG-24 h and initial BW (IBW) were 17 ± 9.8 g/L and 4.1 ± 0.64 kg. The prevalence of FTPI was 29% (220/747 kids). Gastrointestinal complications were the primary cause of death (41%), followed by septicemia (22%) and arthritis (17%). A single penicillin injection reduced preweaning mortality by 55% (10 vs 22%, PEN vs CTR). However, results suggest that such a decline was mainly driven by the improved survival rates among FTPI kids, which increased by 19% (from 62% in CTR-FTPI to 82% in PEN-FTPI), as opposed to an 8% increase among STPI kids (from 85% in CTR-STPI to 93% in PEN-STPI). Additionally, the odds of mortality ≤ 42 d old were threefold higher in the CTR-FTPI group when compared to both the CTR-STPI and PEN-FTPI groups, suggesting a potential parity between STPI and PEN for mortality rate reduction. Taken together, the results indicate that although metaphylactic antibiotics can halve preweaning mortality, similar improvements are likely to be achieved via increased STPI rates. Furthermore, by targeting metaphylactic interventions to high-risk groups (i.e., those displaying signs of inadequate colostrum intake and/or low birth BW), farmers could reduce treatment costs and mitigate AMR risks. While these findings carry considerable weight for commercial dairy goat practices, their applicability to other systems (i.e., extensive, semi-intensive, mohair, meat systems) warrants further investigation., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Current research efforts require a broad range of immune reagents, but those available for pigs are limited. The goal of this study was to generate priority immune reagents for pigs and pipeline them for marketing. Our efforts were aimed at the expression of soluble swine cytokines and the production of panels of monoclonal antibodies (mAbs) to these proteins. Swine interleukin-17A (IL-17A) and Interferon-gamma (IFNγ) recombinant proteins were produced using yeast expression and used for monoclonal antibody (mAb) production resulting in panels of mAbs. We screened each mAb for cross-species reactivity with orthologs of IL-17A or IFNγ and checked each mAb for inhibition by other related mAbs, to assign mAb antigenic determinants. For porcine IL-17A, the characterization of a panel of 10 mAbs identified eight different antigenic determinants; interestingly, most of the mAbs cross-reacted with the dolphin recombinant ortholog. Likewise, the characterization of a panel of nine anti-PoIFNγ mAbs identified four different determinants; most of the mAbs cross-reacted with dolphin, bovine, and caprine recombinant orthologs. There was a unique reaction of one anti-PoIFNγ mAb that cross-reacted with the zebrafish recombinant ortholog. The αIL-17A mAbs were used to develop a quantitative sandwich ELISA detecting the yeast expressed protein as well as native IL-17A in stimulated peripheral blood mononuclear cell (PBMC) supernatants. Our analyses showed that phorbol myristate acetate/ionomycin stimulation of PBMC induced significant expression of IL-17A by CD3+ T cells as detected by several of our mAbs. These new mAbs expand opportunities for immunology research in swine., Competing Interests: JLa and YS are employed by Kingfisher Biotech, Inc., St. Paul, MN, USA. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer SW has declared a past co-authorship with one of the author JL to the handling editor at the time of review., (Copyright © 2022 Manirarora, Walker, Patil, Renukaradhya, LaBresh, Sullivan, Francis and Lunney.)

Trehalose phosphate synthase (TPS), a key enzyme in trehalose synthesis, is not present in mammals but critical to the viability of a wide range of lower organisms. However, almost nothing is known about the function of Hc-TPS (GT1-TPS structural domain protein from Haemonchus contortus ). In this study, Hc-TPS gene was cloned and the recombinant protein (rHc-TPS) was expressed and purified. The quantitative real-time PCR (qPCR) results showed that Hc-TPS was transcribed at different stages of H. contortus , with higher levels of transcription at the molting and embryo stages. Immunofluorescence analysis showed that Hc-TPS was widely distributed in adults, but the expression was mainly localized on the mucosal surface of the intestine as well as in the embryos of female worms. The impacts of rHc-TPS on peripheral blood mononuclear cell (PBMC) proliferation, nitric oxide (NO) generation, transcriptional expression of cytokines, and related pathways were examined by co-incubating rHc-TPS with goat PBMCs. The results showed that rHc-TPS significantly inhibited PBMC proliferation and NO secretion in a dose-dependent manner. We also found that rHc-TPS activated the interleukin (IL)-10/signal transducer and activator of transcription 3/suppressor of cytokine signaling 3 (IL-10/STAT3/SOCS3) axis and significantly promoted SOCS3 expression, while inhibiting interferon-gamma (INF-γ), IL-4, IL-9, and IL-2 pathways. Our findings may contribute to understanding the immune evasion mechanism for the parasite during host-parasite interactions and also help to provide ideas for discovering new drug targets., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Wen, Aleem, Aimulajiang, Chen, Liang, Song, Xu, Li and Yan.)

Peste des petits ruminants (PPR) is an acute transboundary infectious viral disease of small ruminants, mainly sheep and goats. Host susceptibility varies considerably depending on the PPR virus (PPRV) strain, the host species and breed. The effect of strains with different levels of virulence on the modulation of the immune system has not been thoroughly compared in an experimental setting so far. In this study, we used a multi-omics approach to investigate the host cellular factors involved in different infection phenotypes. Peripheral blood mononuclear cells (PBMCs) from Saanen goats were activated with a T-cell mitogen and infected with PPRV strains of different virulence: Morocco 2008 (high virulence), Ivory Coast 1989 (low virulence) and Nigeria 75/1 (live attenuated vaccine strain). Our results showed that the highly virulent strain replicated better than the other two in PBMCs and rapidly induced cell death and a stronger inhibition of lymphocyte proliferation. However, all the strains affected lymphocyte proliferation and induced upregulation of key antiviral genes and proteins, meaning a classical antiviral response is orchestrated regardless of the virulence of the PPRV strain. On the other hand, the highly virulent strain induced stronger inflammatory responses and activated more genes related to lymphocyte migration and recruitment, and inflammatory processes. Both transcriptomic and proteomic approaches were successful in detecting viral and antiviral effectors under all conditions. The present work identified key immunological factors related to PPRV virulence in vitro ., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Eloiflin, Auray, Python, Rodrigues, Seveno, Urbach, El Koulali, Holzmuller, Totte, Libeau, Bataille and Summerfield.)

Maedi-Visna-like genotype A strains and Caprine arthritis encephaltis-like genotype B strains are small ruminant lentiviruses (SRLV) which, for incompletely understood reasons, appear to be more virulent in sheep and goats, respectively. A 9-month in vivo infection experiment using Belgian genotype A and B SRLV strains showed that almost all homologous (genotype A in sheep; genotype B in goats) and heterologous (genotype A in goats; genotype B in sheep) intratracheal inoculations resulted in productive infection. No differences in viremia and time to seroconversion were observed between homologous and heterologous infections. Higher viral loads and more severe lesions in the mammary gland and lung were however detected at 9 months post homologous compared to heterologous infection which coincided with strongly increased IFN-γ mRNA expression levels upon homologous infection. Pepscan analysis revealed a strong antibody response against immune-dominant regions of the capsid and surface proteins upon homologous infection, which was absent after heterologous infection. These results inversely correlated with protection against virus replication in target organs and observed histopathological lesions, and thus require an in-depth evaluation of a potential role of antibody dependent enhancement in SRLV infection. Finally, no horizontal intra- and cross-species SRLV transmission to contact animals was detected.

Background: The liver fluke Fasciola gigantica secretes excretory-secretory proteins during infection to mediate its interaction with the host. In this study, we investigated the immunomodulatory effects of a recombinant tegumental calcium-binding EF-hand protein 4 of F. gigantica (rFg-CaBP4) on goat monocytes., Methods: The rFg-CaBP4 protein was induced and purified by affinity chromatography. The immunogenic reaction of rFg-CaBP4 against specific antibodies was detected through western blot analysis. The binding of rFg-CaBP4 on surface of goat monocytes was visualized by immunofluorescence assay. The localization of CaBP4 within adult fluke structure was detected by immunohistochemical analysis. The cytokine transcription levels in response to rFg-CaBP4 were examined using ABI 7500 real-time PCR system. The expression of the major histocompatibility complex (MHC) class-II molecule (MHC-II) in response to rFg-CaBP4 protein was analyzed using Flow cytometry., Results: The isopropyl-ß-D-thiogalactopyranoside-induced rFg-CaBP4 protein reacted with rat sera containing anti-rFg-CaBP4 polyclonal antibodies in a western blot analysis. The adhesion of rFg-CaBP4 to monocytes was visualized by immunofluorescence and laser scanning confocal microscopy. Immunohistochemical analysis localized native CaBP4 to the oral sucker, pharynx, genital pore, acetabulum and tegument of adult F. gigantica. Co-incubation of rFg-CaBP4 with concanavalin A-stimulated monocytes increased the transcription levels of interleukin (IL)-2, IL-4, interferon gamma and transforming growth factor-β. However, a reduction in the expression of IL-10 and no change in the expression of tumor necrosis factor-α were detected. Additionally, rFg-CaBP4-treated monocytes exhibited a marked increase in the expression of the major histocompatibility complex (MHC) class-II molecule (MHC-II) and a decrease in MHC-I expression, in a dose-dependent manner., Conclusions: These findings provide additional evidence that calcium-binding EF-hand proteins play roles in host-parasite interaction. Further characterization of the immunomodulatory role of rFg-CaBP4 should expand our understanding of the strategies used by F. gigantica to evade the host immune responses.

Previously, it was found that several proteins of Haemonchus contortus were involved in the stimulation of the host immune system. However, the information about the selection of superlative antigens with immunogenic efficacies on host DCs is lacking. In the current study, the stimulatory effects of five recombinant proteins (elongation factor-1α, arginine kinase, ES-15, ES-24, and ADP-ribosylation factor 1) of H. contortus on the maturation of goat monocyte-derived dendritic cells (md-DCs) were reported. Recombinant proteins were purified separately in E. coli expression and incubated with isolated goat peripheral blood mononuclear cells (PBMC). Immunofluorescence assay (IFA) results confirmed the binding of these molecules to the md-DC's surface as compared to control groups. In the flow cytometry analysis, recombinant proteins induced md-DC stimulation via the up-regulation of the expression of the costimulatory molecule (CD80) and MHC-II. Quantitative RT-PCR data showed a significant increase in the expression of specific genes of the WNT and toll-like receptor (TLR) signaling pathways. The result of ELISA indicated the higher levels of cytokine (IL-10, IL-12, IFN-γ, and TNF-α) secretion in the md-DC compared to the negative (pET-32a His-Tag) and blank (PBS) control groups. The data gives valuable support in the selection of potential antigens for future studies on the immunomodulation of the host against the infection of H. contortus., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The major functions of γδ T cells in mammals overlap with those of αβ T cells but differ in that γδ T cells are rapid responders and see different types of antigens. While γδ T cells have been shown to be a major population of circulating lymphocytes in artiodactyl species such as cattle, sheep, and pigs, less is known about these cells in goats, an important agricultural species. We have recently shown that WC1, a γδ T cell-specific family of hybrid pattern recognition receptors/co-receptors, is a multigenic family in goats expanded beyond what occurs in cattle. This study was conducted to address some of the limitations of previous studies in determining the proportions of γδ T cells, WC1 + γδ T cells as well as the WC1.1 + and WC1.2 + subpopulations in blood and to evaluate their responses to various pathogens. Previously, the proportion of caprine γδ T cells was determined using a monoclonal antibody (mAb) 86D that we show here does not react with all γδ T cells thereby underestimating their contribution to the lymphocyte population. Using a mAb reactive with the TCRδ constant region we found the proportion of γδ T cells in blood was not significantly less than that of either CD4 or CD8 T cells and did not decrease with age after 6 months. γδ T cells that expressed WC1 ranged from ~20 to 85% of the total γδ T cells. Less than half of those were classified as WC1.1 + or WC1.2 + by mAb staining thus indicating a third major WC1 + population. We found that naïve γδ T cells proliferated in cultures of PBMC stimulated with antigens of Leptospira or Mycobacterium avium paratuberculosis (MAP) more than they did in control medium cultures or in those stimulated with M. bovis BCG antigens and that the responding γδ T cells included both WC1 + and WC1 - cells. In ex vivo PMA/ionomycin-stimulated cultures of WC1 - γδ T cells but not WC1 + cells produced both IL-17 and IFNγ. In longterm cultures with Leptospira or MAP both WC1 - and WC1 + cells proliferated but only WC1 - γδ T cells produced IL-17. In conclusion, goats have a substantial number of WC1 - and WC1 + γδ T cells in PBMC that do not decrease with animal age after 6 months; both populations respond to bacterial antigens as naïve cells but in these cultures only the WC1 - γδ cells produc IL-17 and IFNγ ., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Mastitis represents one of the major economic and health threats to the livestock sector associated with reduction in milk quality, loss of production and is a major reason for culling. Somatic cell score (SCS) is used as a criterion in breeding programmes to select cows genetically less susceptible to mastitis. The relevance of SCS as a predictor of udder health and susceptibility to mastitis is still untested in goats. In this study, two lines of French Alpine goats selected for extreme breeding values for somatic cell scores, one line with high SCS (HSCS) and the other with low SCS (LSCS), were used to test the hypothesis that the mammary response and function differed between the lines. The aim of the present study was to investigate differences in the early immune response in caprine mammary gland tissues challenged with Staphylococcus aureus, one of the main pathogens responsible for the intra-mammary infection in small ruminants, using transcriptomic and histopathology analyses. The comparison between HSCS and LSCS goat lines, showed differences in the response at the histological level for inflammation, presence of neutrophils and micro-abscess formation, and at the molecular level in the expression of CXCL8, IL-6, NFKBIZ and IL-1β. CXCL8 and CXCL2 genes, which showed a higher level of expression in the experimentally infected HSCS line. The molecular data and histopathology both suggested that following S. aureus infection, mobilization, recruitment, infiltration, and chemotaxis of neutrophil, leads to a more severe inflammation in the HSCS compared to LSCS animals. Our results represent an initial basis for further studies to unravel the genetic basis of early mastitis inflammatory responses and the selection of dairy animals more resistant to bacterial mastitis., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Goats and cattle diverged 30 million years ago but retain similarities in immune system genes. Here, the caprine T cell receptor (TCR) gene loci and transcription of its genes were examined and compared to cattle. We annotated the TCR loci using an improved genome assembly (ARS1) of a highly homozygous San Clemente goat. This assembly has already proven useful for describing other immune system genes including antibody and leucocyte receptors. Both the TCRγ (TRG) and TCRδ (TRD) loci were similarly organized in goats as in cattle and the gene sequences were highly conserved. However, the number of genes varied slightly as a result of duplications and differences occurred in mutations resulting in pseudogenes. WC1 + γδ T cells in cattle have been shown to use TCRγ genes from only one of the six available cassettes. The structure of that Cγ gene product is unique and may be necessary to interact with WC1 for signal transduction following antigen ligation. Using RT-PCR and PacBio sequencing, we observed the same restriction for goat WC1 + γδ T cells. In contrast, caprine WC1 + and WC1 - γδ T cell populations had a diverse TCRδ gene usage although the propensity for particular gene usage differed between the two cell populations. Noncanonical recombination signal sequences (RSS) largely correlated with restricted expression of TCRγ and δ genes. Finally, caprine γδ T cells were found to incorporate multiple TRD diversity gene sequences in a single transcript, an unusual feature among mammals but also previously observed in cattle.

Caprine brucellosis is a chronic, world-wide distributed disease which causes reproductive failure in goats and Brucella melitensis, its causative agent, bears a great zoonotic potential. There is evidence suggesting that some cattle and pigs have an innate ability to resist Brucella infection, but this has not yet been investigated in goats. In this study, we compared caprine macrophages that exhibit extreme restriction and permissiveness to B. melitensis' intracellular growth in vitro. Monocyte derived macrophages (MDMs) from 110 female goats were cultured and challenged in vitro with B. melitensis 16 M. After initial screening, 18 donor goats were selected based on their macrophages ability to restrict or allow bacterial intracellular growth and some elements of humoral and cellular immunity were studied in depth. MDMs that were able to restrict the pathogen's intracellular growth showed enhanced bacterial internalization, although there were no differences between groups in the production of reactive oxygen and nitrogen intermediates following 48 h treatment with heat-killed B. melitensis. Moreover, there were no differences between groups in the level of antibodies reacting with keyhole limpet hemocyanin (natural antibodies, NAbs) or with Brucella LPS antigens (cross-reacting antibodies, CrAbs), although a strong positive correlation between individual levels of IgM NAbs and IgM CrAbs was detected. Altogether, these results represent an initial step in understanding innate primary host response to B. melitensis, and deciphering which mechanisms may determine a successful outcome of the infection in goats., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Arthropod-borne Batai virus (BATV) is an Orthobunyavirus widely distributed throughout European livestock and has, in the past, been linked to febrile diseases in humans. In Germany, BATV was found in mosquitoes and in one captive harbor seal, and antibodies were recently detected in various ruminant species. We have, therefore, conducted a follow-up study in ruminants from Saxony-Anhalt, the most affected region in Eastern Germany. A total of 325 blood samples from apparently healthy sheep, goats, and cattle were tested using a BATV-specific qRT-PCR and SNT. Even though viral RNA was not detected, the presence of antibodies was confirmed in the sera of all three species: sheep (16.5%), goats (18.3%), and cattle (41.4%). Sera were further analyzed by a glycoprotein Gc-based indirect ELISA to evaluate Gc-derived antibodies as a basis for a new serological test for BATV infections. Interestingly, the presence of neutralizing antibodies was not directly linked to the presence of BATV Gc antibodies. Overall, our results illustrate the high frequency of BATV infections in ruminants in Eastern Germany.

Currently, brucellosis is a reemerged zoonotic infectious disease with an increased incidence in recent years. A simple, rapid and sensitive method for diagnosing brucellosis can help to reduce medical burden and economic loss. Previously, a multiple epitope recombinant protein was constructed based on linear B-cell epitope prediction tools. In this study, the recombinant protein was used as an antigen to study the immune response produced by immunized mice, and goat serum was used to verify its diagnostic accuracy. The production of antibodies was successfully induced in the vaccinated mice. Flow cytometric analysis revealed that the percentage of CD4 + , CD8 + and the CD4+/CD8+ ratios were increased by T cell subsets in mouse splenocytes, indicating that the recombinant protein induced a strong immune response had strong immunoreactivity. Using indirect ELISA, the recombinant protein correctly diagnosed positive and negative brucellosis samples. Compared with the whole bacterial antigen, the recombinant protein had a weaker sensitivity but a stronger specificity. Animal experiments showed that the recombinant protein had good antigenicity, and indirect ELISA indicates that it can be used as an antigen to diagnose brucellosis. Therefore, the recombinant protein is a potential candidate antigen for brucellosis vaccine development and serological diagnosis., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Newborn ungulates depend on the timely supply of colostrum containing sufficient immunoglobulins to obtain passive immunity against disease. Brix refractometry enables a rapid on-farm estimation of colostrum quality and has been intensively studied in bovines. However, the suitability of Brix refractometers for assessing colostrum quality in goats and ewes has been scarcely evaluated. The present study compared bovine, caprine, and ovine colostrum quality estimation using an optical Brix refractometer. In addition, between-species variations in the relationships between Brix values and colostrum constituents (IgG, fat, protein, and lactose) and the accuracy of Brix refractometry at different cutoff values were evaluated by a receiver operating characteristic curve analysis. We measured the Brix value and contents of IgG, fat, protein, and lactose in 324 colostrum samples (108 cows, 116 does, and 100 ewes). Thresholds for classification of good colostrum quality (as determined by ELISA) were set at 50 mg IgG/mL in cows and 20 mg/mL in does and ewes. Bovine colostrum showed the greatest IgG concentrations compared with caprine and ovine colostrum. Fat and protein content was higher in sheep colostrum compared with the other species, whereas the highest lactose concentrations were detected in goat colostrum. Brix values ranged from 11.4 to 34.6% (22.1 ± 4.2%; mean ± standard deviation), 15.4 to 40.0% (28.5 ± 6.8%), and 8.8 to 39.8% (21.6 ± 5.3%) in bovine, ovine, and caprine colostrum, respectively. In all 3 species, Brix was highly correlated with IgG and protein concentrations (cows, r = 0.83 and 0.98; goats, r = 0.83 and 0.89; sheep, r = 0.75 and 0.87). Optimal cutoff points for greatest accuracy of Brix measurements were 19.3% Brix in cows [with 87.1% sensitivity (Se) and 100% specificity (Sp)], 20.7% Brix in does (with 53.5% Se and 100% Sp), and 26.5% Brix in ewes (with 75% Se and 91.3% Sp). In conclusion, Brix refractometry is an acceptable tool for on-farm estimations of colostrum quality in does and ewes despite distinct between-species variations in colostrum composition., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Background: Long-term high-concentrate (HC) diet feeding increased bacterial endotoxins, which translocated into the mammary glands of dairy goats and induced inflammatory response. γ-d-Glutamyl-meso-diaminopimelic acid (iE-DAP), bacterial peptidoglycan component, triggered inflammatory response through activating nucleotide oligomerization domain protein 1 (NOD1) signaling pathway. While dietary supplemented with sodium butyrate (SB) relieved inflammatory response and improved animal health and production. To investigate the effects and the mechanisms of action of SB on the inflammatory response in the mammary glands of dairy goats fed HC diet, 12 Saanen dairy goats were randomly assigned into HC group and SB regulated (BHC) group., Results: The results showed that SB supplementation attenuated ruminal pH decrease caused by HC diet in dairy goats resulting in a decrease of proinflammatory cytokines and iE-DAP plasma concentration and the mRNA expression of NOD1 and other inflammation-related genes. The protein levels of NOD1, NF-κB p65 and NF-κB pp65 were decreased by the SB supplementation. The expression of histone deacetylase 3 (HDAC3) was also inhibited by the SB supplementation. Meanwhile, the chromatin compaction ratios and DNA methylation levels of NOD1 and receptor-interacting protein 2 (RIP2) of BHC group were upregulated., Conclusion: Collectively, the SB supplementation mitigated the inflammatory response in the mammary glands of dairy goats during HC-induced subacute ruminal acidosis (SARA) by inhibiting the activation of the NOD1/NF-κB signaling pathway through the decrease of the iE-DAP concentration in the rumen fluid and plasma and HDAC3 expression. DNA methylation and chromatin remodeling also contributed to the anti-inflammatory effect of SB. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Goats are important food animals and are disseminated globally because of their high adaptability to varying environmental conditions and feeding regimes that provide them with a comparative advantage. Productivity is impacted by infectious diseases; this then contributes to societal poverty, food insecurity, and international trade restrictions. Since γδ T cells have been shown to have vital roles in immune responses in other mammals we reviewed the literature regarding what is known about their functions, distribution in tissues and organs and their responses to a variety of infections in goats. It has been shown that caprine γδ T cells produce interferon-γ and IL-17, are found in a variety of lymphoid and nonlymphoid tissues and constitute a significant population of blood mononuclear cells. Their representation in tissues and their functional responses may be altered concomitant with infection. This review summarizes caprine γδ T cell responses to Brucella melitensis, Fasciola hepatica, Mycobacterium avium paratuberculosis, caprine arthritis encephalitis virus (CAEV), and Schistosoma bovis in infected or vaccinated goats. Caprine γδ T cells have also been evaluated in goats infected with M. caprae, Ehrilichia ruminantium, Haemonchus contortus and peste des petits ruminants (PPR) virus but found to have an unknown or limited response or role in either protective immunity or immunopathogenesis in those cases., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The objectives of the study were to determine the dose-dependent effects of active immunization against inhibin α-subunit (AIINHA) on ovarian dynamics, concentrations of progesterone (P 4 ), pregnancy rate (PR), embryonic and fetal losses (EFL), and prolificacy during the non-breeding season when there was imposing of a progestin-based treatment regimen to induce estrus in Beetal goats. Goats (n = 30) were randomly assigned into following groups: 1) saline (G-CON-0 mg; n = 10), 2) small dose (G-AIINHA-0.5 mg; n = 10), and 3) large dose (G-AIINHA-1 mg; n = 10). The primary administration of inhibin immunogen was administered at Day -48, followed by another administration at Day -20, and subsequently there was induction of estrus using a progestin based treatment regimen that included a single administration of progestin-containing sponge and PGF 2α at Day -8. The sponge was removed, and GnRH was administered at Day -3 followed by breeding (Day 0) at standing estrus. Results indicated mean diameter of the follicles, size of pre-ovulatory follicles and corpora lutea, and post-breeding P 4 concentrations were greater (P < 0.05) in the goat does of the G-AIINHA-0.5 than G-CON-0 group. The PR, and EFL, however, did not differ (P> 0.05) among groups, whereas prolificacy rate was greater (P = 0.04) in goat does of the G-AIINHA-0.5 than G-CON-0 groups. The data from this study indicate G-AIINHA-0.5 is the recommended dose of inhibin immunogen to enhance the reproductive performance during non-breeding season in Beetal goats when estrus is induced using a progestin-based treatment regimen., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Peripheral blood from healthy sheep (n = 3) and goats (n = 3) were employed to establish an efficient method for simultaneous isolation of peripheral blood mononuclear cells (PBMCs) and neutrophils and to standardize protocols for monocyte purification and generation of monocyte-derived macrophages (MDMs). In both species, a significantly enriched population of PBMCs, with higher purity and number of cells determined by flow cytometry, was achieved when processing through a density gradient a mixture of buffy-coat and red blood cell layer (RBC) in comparison to the use of just the buffy-coat (p <  0.05). Neutrophils could be subsequently isolated from the layer, located underneath PBMCs fraction with significant higher purity rates, higher than 85 % determined by flow cytometry, than those obtained with protocols without density gradients (< 60 %) (p <  0.05). This technique would allow the isolation of both cell populations from the same sample of blood. A pure cell population of monocytes, CD14 + cells, was purified from PBMCs when using immunomagnetic columns, which allow for 17 % (nº monocytes/nº PBMCs) of yield and high percentages of expression of CD14 + (88 %), MHC-II + (91.5 %) and CD11b + (94 %) established by flow cytometry. On the other hand, the classical and non-expensive purification of monocytes from PBMCs based on the adherence capacity of the former, allowed significantly lower yield of monocytes (4.6 %), with percentages of surface markers expression that dropped to 35 %, 65 % and 55 %, respectively (p < 0.001), suggesting the isolation of a mixed population of cells. The addition of GM-CSF to the culture, at concentration from 25 to 125 ng/mL, enhanced proportionally the number of MDMs generated compared to the absence of supplementation or the use of autologous serum from 5% to 20 %. However, purification of monocytes through the adherence method achieved higher yields of MDMs than those isolated through immunomagnetic columns in both species (p <  0.001). Under the conditions of this study, the use of centrifugation in density gradients allow for the simultaneous purification of PBMCs and neutrophils, with high purity of both populations, from the same sample of blood. The isolation of monocytes could be subsequently achieved through two different methods, i.e. based on immunomagnetic columns or adherence. The preference between both methods would depend on the necessities of the experiment, the initial sample with high purity of monocytes or a final population of MDMs required., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Assessment of immune fitness is valuable in many aspects of livestock management and research. Determining immune consequences of selection for increased disease resistance or inhabiting various environments or climates can lead to different management decisions. The ability to measure immune responses due to different diets, pregnancy status, or aging will increase insight about how these factors contribute to overall immune health. The main objective of these experiments was to adapt a methodology used in cattle and pigs to measure both the humoral and cell-mediated immune response in sheep and goats. The route of administration of two antigens, Candida albicans and hen egg white lysozyme, were compared in sheep to determine differences in antibody or cell-mediated immune response. Subcutaneous injection produced a larger (P < 0.001) cell-mediated response compared to intramuscular injection. Inoculation in the axillary space produced a larger (P = 0.0031) antibody response compared to neck region. Finally, methodology was confirmed in goats. Complete blood cell counts were compared and lymphocytes were highest in low cell-mediated responders while eosinophils were highest in average antibody-mediated responders. This work provides a means to measure immune fitness in sheep and goats allowing for future experiments examining environmental or genetic effects on the immune response., (Published by Elsevier B.V.)

Objective: To understand the origin of extremely high gonadotropin levels in a perimenopausal woman. Methods: A 52-year-old woman with a 2 months of amenorrhea followed spontaneous menstrual cycles recovery was referred to our outpatient clinic with elevated follicle-stimulating hormone (FSH, 483 mUI/ml), luteinizing hormone (LH, 475 mUI/ml) and prolactin (PRL, 173 ng/ml). She was known to take levosulpiride. The gonadotropin levels did not fit with the clinical features. Results: A gonadotroph tumor was ruled out. Further analysis confirmed constantly high FSH, LH and PRL levels. The measurements were repeated using different analytical platforms with different results. After serial dilutions, nonlinearity was present suggesting an immunoassay interference. After post-polyethylene glycol recovery, hormone levels appeared in the normal range. Anti-goat antibodies were recognized in the serum of the patient. Conclusions: This case report shows a case of falsely abnormal high gonadotropin and PRL levels in a woman during menopause transition. In the clinical practice the evaluation of gonadotropin profile is not recommended at this age, but the abnormal levels stimulated further evaluation. An interference in the assay due to anti-goat antibodies resulted in abnormally high level of FSH and LH. A strict collaboration between clinicians and the laboratory is needed, when laboratory findings do not correspond to clinical findings.

On-farm assessment of caprine colostrum quality is important for goat farmers; the ability to quickly recognize whether colostrum is suitable to feed to kids helps achieve successful passive transfer of immunity. The study compared the use of optical and digital Brix refractometers and a hydrometer against the international gold standard radial immunodiffusion (RID), using both fresh and frozen samples. A locally available ELISA methodology was included for comparison. A total of 300 samples were collected from 2 farms (farm 1: n = 157, collected by research staff within 24 h of parturition; farm 2: n = 143, collected by the farmer within 12 h of parturition). Farm 1 provided doe age for a subset of samples (n = 86). Samples were tested fresh and then frozen for shipment and repeated testing. Specific gravity was measured using a hydrometer in a subset of samples (n = 22) from farm 2. Because no gold standard thresholds are currently available for caprine colostrum, RID-derived values of 30, 40, and 50 g/L IgG were used as potential "good quality" thresholds. Pearson (ρ) and Lin's concordance correlation coefficients (CCC) were calculated for comparison of methods. Optimum thresholds were established maximizing the Youden index and minimizing the "distance closest to the top left corner" of the receiver operator characteristic curves. Brix values were correlated with RID (optical Brix, fresh: ρ = 0.73; digital Brix, fresh: ρ = 0.71; digital Brix, frozen: ρ = 0.76) and with each other (range: ρ = 0.93 to 0.99; CCC = 0.91 to 0.99). Specific gravity measured by the hydrometer yielded a strong relationship with RID (ρ = 0.83) and with Brix values (range: ρ = 0.88 to 0.90). The ELISA method was not correlated with Brix methods (range: ρ = 0.02 to 0.09) or RID (ρ = 0.20). Depending on the colostrum IgG threshold, the hydrometer yielded high Youden indices (range: 0.78 to 0.93) and low distance closest to the top left corner criteria (0 to 0.05) at a threshold of 1.047 specific gravity. For all RID IgG thresholds, the best Brix threshold (regardless of type or whether the sample was fresh or frozen) was 18 or 19%, with the highest Youden indices (range: 0.47 to 0.61) and lowest distance to the top left corner criteria (range: 0.09 to 0.16); however, we recommend 19%, because this reduces the potential of feeding poor-quality colostrum. The ELISA method was the poorest predictor of colostrum concentration. Age was not found to affect colostrum quality; however, the sample size of this subset was small. Hydrometers are inexpensive and easy to use, whereas Brix methods use only a small amount of colostrum; we suggest that either method could be used on-farm., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).)

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.

The presence of antibodies to bluetongue virus (BTV) and the viral antigen is reported recently from the Andaman and Nicobar Islands, a group of islands at the juncture of the Bay of Bengal and the Andaman Sea. A retrospective study was conducted to investigate the presence of neutralizing antibodies to different BTV serotypes in the seroconverted goats of the Islands. Thirty six samples out of 186 serum samples tested were selected on the basis of high antibody titre as predicted in an indirect ELISA. Each of the selected serum samples was used for neutralization of six BTV serotypes (BTV-1, BTV-2, BTV-9, BTV-10, BTV-16 and BTV-23), the most commonly reported serotypes in India. Out of 36 serum samples used in the neutralization study, neutralizing antibodies could be determined in 15 samples. The neutralizing antibodies to BTV-10 were found in more number of the serum samples followed by BTV-1, BTV-2 and BTV-23 and BTV-9 and BTV-16. Many of the serum samples could neutralize more than one BTV serotypes indicating possible widespread superinfections by multiple BTV serotypes in goats in the Islands. Majority of the serum samples used in the neutralization study could not neutralize any of the six BTV serotypes commonly reported from India indicating possible circulation of other BTV serotypes yet to confirm. The present study reveals circulation of multiple BTV serotypes in Andaman and Nicobar Islands where there was no such report available earlier. The findings are laudable as the baseline information for further investigations to identify and characterize the virus and competent vectors and for implementing appropriate suitable control strategies for bluetongue in the Islands and the nearby territories.

Strongylida are gastrointestinal nematodes (GIN) of greatest importance in small ruminants throughout the world. Differences in resistance and resilience to GIN among goat breeds were reported. This study aims to investigate the mechanism underlying the breed-associated differences using a cosmopolitan (Alpine, AB) and an autochthonous (Nera di Verzasca, NV) goat breed. At first, fifteen goats from the same herd (NV = 7, AB = 8) at day 0 were infected with infective larvae (L3) of mixed GIN. From the 15th day post-infection (DPI), individual parasite egg excretion (faecal egg counts, FEC) was performed on all goats, once per week, until the 63rd DPI. Afterwards, in goats under field conditions (30 AB and 30 NV reared on the same farm), individual faecal and blood samples were collected; FEC-specific antibody and PCV levels were explored. In goats with experimental GIN infection, mean eggs per gram of faeces (EPG) values were consistently lower in NV goats. In goats with natural GIN infection, EPG and prevalence values showed high variability in both breeds; among individual variables, breed had a significant influence on EPG. Further, PCV and anti-T. circumcincta IgA levels were influenced by the breed. Lower PCV values were also associated with higher strongyle EPG in AB goats, and anti-T. circumcincta IgA levels were influenced by both strongyle EPG and breed, with IgA levels being higher in AB vs. NV goats and positively associated with EPG. Neither EPG nor breed had any influence on IgE levels. Both studies on experimental and natural infection confirmed that goats of NV are more resistant to infection with gastrointestinal nematodes.

Algae are a rich source of bioactive compounds and health properties that have been narrowly explored in goat production systems. The aim of this study was to determine the effect of feeding diets supplemented with Sargassum spp. on antioxidant status and immune parameters in goat kids. The diets were as follows: control (basal diet without alga), Sargassum spp. 2.5% (Ss2.5), and Sargassum spp. 5% (S5) fed over a 70-day period. A total of 11 body tissues, intestinal mucus, and blood serum were sampled at necropsy. Protein content, superoxide dismutase (SOD), catalase (CAT), myeloperoxidase (MPO), lysozyme, and anti-protease activities were determined, as well as immunoglobulin A (IgA) and immunoglobulin G (IgG). The results indicated that Sargassum spp. supplementation increased protein content in six tissues. Antioxidant activities (SOD and CAT) and immune-related (lysozyme, MPO, and anti-protease) activities were statistically higher (P < 0.05) in Sargassum spp. groups compared with control in several tissues, intestinal mucus, and serum. Imunoglobulin A levels in intestinal mucus were higher (P < 0.05) in Sargassum spp.-supplemented groups than the control group. In conclusion, diet supplementation of Sargassum spp. improves the antioxidant status and enhances the immune parameters in goats. Sargassum spp. dietary supplementation is proposed as strategy to strengthen antioxidant status and stimulate the immune system, which helps in the control of opportunistic pathogens in goats.

Aims: Most breeds of goat are more susceptible to nematode infection than sheep, and this appears to be a consequence of less effective immune responses. Several papers have considered the effectiveness of eosinophils and immunoglobulin A (IgA) in goats but differences in the induction of responses have not been studied in the same detail. The aim of this study was to look at the induction of eosinophil and IgA responses in Boer goats reared indoors under intensive conditions., Methods and Results: The goats were experimentally infected with a low dose of 2400 Haemonchus contortus, Trichostrongylus spp. and Oesophagostomum spp. at a 6:1:1 ratio. Faecal egg counts (FEC), packed cell volume (PCV), IgA activity against third-stage larvae and peripheral eosinophilia were measured twice a week for eight weeks. The infection generated an IgA response but did not significantly increase peripheral eosinophilia in the 25 infected kids compared with the 4 control animals. FEC was not associated with IgA activity or eosinophilia., Conclusion: A detailed analysis of IgA and eosinophil responses to deliberate nematode infection in Boer goats showed that there was an increase in nematode-specific IgA activity but no detectable eosinophil response. In addition, there was no association between increased IgA activity or eosinophilia with egg counts and worm burdens. These suggest that IgA and eosinophils do not act to control nematode infection in goats., (© 2020 John Wiley & Sons Ltd.)

Haemonchus contortus has evolved highly integrated and sophisticated mechanisms to promote coexistence with hosts. The excretory-secretory (ES) products generated by this parasite contribute to the regulation of the host immune response to facilitate immune evasion and induce chronicity, but the proteins responsible for this process and the exact cellular mechanisms have yet to be defined. In this study, we identified 114 H. contortus ES proteins (HcESPs) interacting with host T cells and 15 T cell binding receptors via co-immunoprecipitation and shotgun liquid chromatography-tandem mass spectrometry analysis. Based on bioinformatics analysis, we demonstrated that HcESPs could inhibit T cell viability, induce cell apoptosis, suppress T cell proliferation and cause cell cycle arrest. Furthermore, the stimulation of HcESPs exerted critical control effects on T cell cytokine production profiles, predominantly promoting the secretion of interleukin (IL)-10, IL-17A and transforming growth factor-β1 and inhibiting IL-2, IL-4 and interferon-γ production. Collectively, these findings may provide insights into the interaction between ES proteins and key host effector cells, enhancing our understanding of the molecular mechanism underlying parasite immune evasion and providing new clues for novel vaccine development.

Suppression of ubiquitin proteasome pathway (UPP) and stimulation of caspase-3 are involved in neurodegeneration. Can UPP activators and caspase-3 inhibitors ameliorate neurodegeneration? Here, we found a novel neuronal cell death accompanied with UPP activation and caspase-3 inhibition. Recently, plasmalemmal neuron-specific enolase (NSE) has been identified as one of membrane targets of 15-deoxy-Δ 12,14 -prostaglandin J 2 (15d-PGJ 2 ). 15d-PGJ 2 induces neuronal apoptosis via activating caspase-3 and inactivating UPP, whereas the anti-NSE antibody inactivated caspase-3, activated UPP, and caused neuronal cell death. The anti-NSE antibody activated caspase-1 (pyroptosis marker), but not condense chromatin (apoptosis marker). The anti-NSE antibody declined intracellular level of ATP, which is not altered in pyroptosis. The intracellular level of calcium is elevated in necrosis and pyroptosis, but its chelator did not ameliorate the neurotoxicity of anti-NSE. Thiol antioxidants such as N-acetyl cysteine and glutathione reduced the neurotoxicity of 15d-PGJ 2 but enhanced that of the anti-NSE antibody. The anti-NSE antibody incorporated propidium iodide into neurons through the disrupted plasma membrane, which are not observed in ferroptosis and autophagic cell death. Thus, the anti-NSE antibody induced neuronal cell death in a novel fashion distinguished from necrosis, necroptosis, apoptosis, pyroptosis, ferroptosis, and autophagic cell death.

Background: Tick distribution in Sweden has increased in recent years, with the prevalence of ticks predicted to spread towards the northern parts of the country, thus increasing the risk of tick-borne zoonoses in new regions. Tick-borne encephalitis (TBE) is the most significant viral tick-borne zoonotic disease in Europe. The disease is caused by TBE virus (TBEV) infection which often leads to severe encephalitis and myelitis in humans. TBEV is usually transmitted to humans via tick bites; however, the virus can also be excreted in the milk of goats, sheep and cattle and infection may then occur via consumption of unpasteurised dairy products. Virus prevalence in questing ticks is an unreliable indicator of TBE infection risk as viral RNA is rarely detected even in large sample sizes collected at TBE-endemic areas. Hence, there is a need for robust surveillance techniques to identify emerging TBEV risk areas at early stages., Methods: Milk and colostrum samples were collected from sheep and goats in Örebro County, Sweden. The milk samples were analysed for the presence of TBEV antibodies by ELISA and validated by western blot in which milk samples were used to detect over-expressed TBEV E-protein in crude cell extracts. Neutralising titers were determined by focus reduction neutralisation test (FRNT). The stability of TBEV in milk and colostrum was studied at different temperatures., Results: In this study we have developed a novel strategy to identify new TBEV foci. By monitoring TBEV antibodies in milk, we have identified three previously unknown foci in Örebro County which also overlap with areas of TBE infection reported during 2009-2018. In addition, our data indicates that keeping unpasteurised milk at 4 °C will preserve the infectivity of TBEV for several days., Conclusions: Altogether, we report a non-invasive surveillance technique for revealing risk areas for TBE in Sweden, by detecting TBEV antibodies in sheep milk. This approach is robust and reliable and can accordingly be used to map TBEV "hotspots". TBEV infectivity in refrigerated milk was preserved, emphasising the importance of pasteurisation (i.e. 72 °C for 15 s) prior to consumption.

Beta-glucans from yeast can induce trained immunity in in vitro and in vivo models. Intraperitoneal doses of β-glucans in mammals have shown to induce trained immunity, but the training effects of orally administering β-glucans are unknown. Newborn goats are susceptible to infections in the neonatal stage, so the induction of trained immunity could improve animal survival. This study aimed to describe the in vitro effects of immunological training by β-glucan from Debaryomyces hansenii (β-Dh) on caprine monocytes, as well as its in vivo effects using oral doses on newborn goats upon challenge with lipopolysaccharide (LPS). Hence in vitro, goat monocytes trained with β-Dh up-regulated the gene expression of macrophage surface markers (CD11b and F4/80) whereas enhanced cell survival and high phagocytic ability was found upon LPS challenge. In the in vivo experiment, newborn goats stimulated with two doses (day -7 and - 4) of β-Dh (50 mg/kg) and challenged (day 0) with LPS showed an increase in respiratory burst activity, IL-1β, IL-6, and TNFα production in plasma, and transcription of the macrophage surface markers. This study has demonstrated for the first time that trained immunity was induced with oral doses of β-glucan upon LPS challenge in mammals using newborn goats., Competing Interests: Declaration of competing interest The authors have no financial conflicts of interest., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Mammary serum amyloid A3 (M-SAA3) has shown potential in stimulating innate immunity during intramammary infections, at calving and at dryoff. In this study, we produced recombinant caprine M-SAA3 to test its ability to reduce intramammary infections with Staphylococcus aureus, Streptococcus uberis, Streptococcus dysgalactiae, and Escherichia coli, which are all common mastitis-producing pathogens. Recombinant production of M-SAA3 (followed by lipopolysaccharide removal to avoid lipopolysaccharide-nonspecific stimulation of the immune system) was successfully achieved. Mammary serum amyloid A3 stimulated the expression of IL-8 in a dose-dependent manner in primary mammary cultures. Although a direct killing effect on Staph. aureus by M-SAA3 was not detected, this acute phase protein was able to reduce Staph. aureus, Strep. uberis, and Strep. dysgalactiae infections by up to 50% and induced a reduction in E. coli counts of 67%. In general, the best concentration of caprine M-SAA3 for inhibiting infections was the lowest concentration tested (10 μg/mL), although higher concentrations (up to 160 μg/mL) increased its antimicrobial potential against some pathogens., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Contagious caprine pleuropneumonia (CCPP) is a highly contagious infectious disease of goats caused by Mycoplasma capricolum subspecies capripneumoniae (Mccp). CCPP outbreaks usually result in high morbidity and mortality of the affected goats, making this disease a major cause of economic losses to goat producers globally. However, the pathogenesis of CCPP remains unclear. Here, we show that IL-17-driven neutrophil accumulation is involved in the lung damage in CCPP goats. During CCPP development, intense inflammatory infiltrates could be observed in the injured lungs. Specifically, neutrophils were observed to be present within the alveoli. Increased IL-17 release drove the excessive influx of neutrophils into the lung, as IL-17 effectively stimulated the production of neutrophil chemoattractants from lung epithelial cells following Mccp infection. Our data highlight a critical role of IL-17-driven neutrophil accumulation in the pathogenesis of CCPP and suggest that IL-17 may potentially be a useful immunotherapeutic target for the treatment of CCPP., Competing Interests: Declaration of Competing Interest The authors have no conflicts of interest to declare., (Copyright © 2020 Elsevier B.V. All rights reserved.)

While it is now broadly accepted that inter-individual variation in the outcomes of host-pathogen interactions is at least partially genetically controlled, host immunogenetic characteristics are rarely investigated in wildlife epidemiological studies. Furthermore, most immunogenetic studies in the wild focused solely on the major histocompatibility complex (MHC) diversity despite it accounts for only a fraction of the genetic variation in pathogen resistance. Here, we investigated immunogenetic diversity of the Alpine ibex (Capra ibex) population of the Bargy massif, reservoir of a virulent outbreak of brucellosis. We analysed the polymorphism and associations with disease resistance of the MHC Class II Drb gene and several non-MHC genes (Toll-like receptor genes, Slc11A1) involved in the innate immune response to Brucella in domestic ungulates. We found a very low neutral genetic diversity and a unique MHC Drb haplotype in this population founded few decades ago from a small number of individuals. By contrast, other immunity-related genes have maintained polymorphism and some showed significant associations with the brucellosis infection status hence suggesting a predominant role of pathogen-mediated selection in their recent evolutionary trajectory. Our results highlight the need to monitor immunogenetic variation in wildlife epidemiological studies and to look beyond the MHC.

This experiment aimed to evaluate the suitability of glycerol and propylene glycol to reduce microbial count and preserve immune properties in heat-treated goat colostrum. Colostrum samples from 11 goats were each divided into 9 aliquots. Different concentrations (2, 6, 10, and 14%; vol/vol) of either glycerol or propylene glycol were added to the aliquots. Phosphate buffer solution was added to one aliquot, which was set as the control (CG). After the respective additions, all colostrum samples were heat treated at 56°C for 1 h. After cooling, aerobic mesophilic bacteria were cultured. The samples were frozen until free fatty acid, IgG, IgA, and IgM concentrations and chitotriosidase activity were measured. No differences were found in aerobic mesophilic bacteria counts between either 10 or 14% glycerol and propylene glycol additives. These additions reduced bacterial count to a greater extent than CG, and 2 or 6% additions. Colostrum IgG concentration was not affected by either of the additives or their concentrations. The propylene glycol additive reduced IgA and IgM concentrations and chitotriosidase activity, compared with CG. Conversely, glycerol did not affect any of the studied immune variables. In conclusion, glycerol addition to goat colostrum before heat treatment is suitable to enhance bacterial reduction, whereas colostrum immune properties were not affected., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Goat milk (GM), as compared to cow milk (CM), is easier for humans to digest. It also has antioxidant and anti-inflammatory effects and can improve minor digestive disorders and prevent allergic diseases in infants. It is unclear whether GM consumed in pregnant mothers has any protective effects on allergic diseases in infants. In this experimental study with mice, we found GM feeding enhanced immunoglobulin production, antigen-specific (ovalbumin, OVA) immune responses, and phagocytosis activity. The GM-fed mice had an increasing proportion of CD3 + T lymphocytes in the spleen. Splenocytes isolated from these animals also showed significantly increased production of cytokines IFN-γ and IL-10. More importantly, GM feeding during pregnancy and lactation periods can confer protective activity onto offspring by alleviating the airway inflammation of allergic asthma induced by mite allergens. There was a remarkably different composition of gut microbiota between offspring of pregnant mice fed with water or with milk (GM or CM). There was a greater proportion of beneficial bacterial species, such as Akkermansia muciniphila, Bacteroides eggerthii , and Parabacteroides goldsteinii in the gut microbiota of offspring from GM- or CM-fed pregnant mice compared to the offspring of water-fed pregnant mice. These results suggested that improving the nutrition of pregnant mice can promote immunological maturation and colonization of gut microbiota in offspring. This mother-to-child biological action may provide a protective effect on atopy development and alleviate allergen-induced airway inflammation in offspring., (Copyright © 2020 Kao, Wang, Tseng, Wu, Tsai, Hsieh, Chen, Kuo, Liu, Liu and Wang.)

Studies in mice with ablation of Prnp, the gene that encodes the cellular prion protein (PrP C ), have led to the hypothesis that PrP C is important for peripheral nerve myelin maintenance. Here, we have used a nontransgenic animal model to put this idea to the test; namely, goats that, due to a naturally occurring nonsense mutation, lack PrP C . Teased nerve fiber preparation revealed a demyelinating pathology in goats without PrP C . Affected nerves were invaded by macrophages and T cells and displayed vacuolated fibers, shrunken axons, and onion bulbs. Peripheral nerve lipid composition was similar in young goats with or without PrP C , but markedly different between corresponding groups of adult goats, reflecting the progressive nature of the neuropathy. This is the first report of a subclinical demyelinating polyneuropathy caused by loss of PrP C function in a nontransgenic mammal., (© 2019 Norwegian University of Life Sciences. The FASEB Journal published by John Wiley & Sons Ltd on behalf of The FASEB Journal.)

Galectins are glycan-binding proteins that are widely expressed and distributed in mammalian tissues as well as cells of innate and adaptive immune responses. CD4+ T-helper cells differentiate into effector subsets in response to cytokines. T helper 9 cells are one of the recently described subsets of effector T cells that are relatively new and less studied. In this study, galectin domain containing protein from Haemonchus contortus (Hc-GDC) was cloned, expressed in pET32a, and immunoblotting was performed. Localization of recombinant (r)Hc-GDC on outer and inner surface of H. contortus worm and binding with goat Peripheral Blood Mononuclear cells (PBMCs) were performed using immunofluorescence assay. Moreover, effects of rHc-GDC on proliferation, apoptosis, cell migration, and the nitric oxide production in goat PBMCs were evaluated. Furthermore, modulatory effects of rHc-GDC on production of Th1, Th2, and Th9 cells were evaluated by flowcytometry and on interferon gamma, interleukin (IL)-4 and IL-9 were evaluated by quantitative real-time polymerase chain reaction. The results demonstrated that rHc-GDC was successfully cloned, expressed in expression vector as well as in the gut surface of adult H. contortus worm and successful binding with PBMCs surface were observed. Immunoblotting results revealed that rHc-GDC is an important active protein of H. contortus excretory and secretory products. Moreover, the interaction of rHc-GDC with host cells increased the production of Th2, Th9 cells, IL4, IL-9, PBMC proliferation, nitric oxide, and cell migration. No effects of rHc-GDC were observed on PMBC apoptosis, production of Th1 cells, and secretions of IFN- and IL-10 cytokines. These findings indicate that recombinant GDC protein from H. contortus modulates the immune functions of goat PBMCs and has the potential to enhance protective immunity by inducing T helper-9-derived IL-9 in vitro., Competing Interests: Appendix A

The aim of the present study was to define whether elevated rumen-undegradable protein (RUP) and tannin-rich peanut skin (PS) supplementation would affect animal growth performance, average daily gain (ADG), blood metabolites, carcass traits associated with lipogenic and immune-related gene expressions in meat goats grazing winter wheat (WW). Thirty-six Kiko-crossbreed male goats at approximately 6 months of age were blocked by body weight (BW; 25.6 ± 1.1 kg) and randomly assigned to one of the four treatments with two replicates based on a 2 × 2 factorial design. Diets contained PS replacing alfalfa meal (ALM), without or with RUP supplementation. Both PS and ALM were incorporated into grain mix portion of the diet and pelletized, with remaining diets fed ad libitum of WW forage for a period of 51 days. Lipogenic genes examined included SCD, ACLY, YWHAZ, PPIA and FABP4, while immune-related genes examined included ACTB (as a control gene), H3F3A, PPIA, IRF3, STAT2, HERC3 and IFIT3 antibody genes. The meat goats on PS-pellet-supplemented group with or without RUP supplementation grew 38.5% faster ADG (p < .001) when compared to control-supplemented group. When goats received PS diet, empty body weight, hot carcass, cold carcass, shoulder, hind shank, rack, loin and fat thickness were greater (p < .05) than control diet. Animals on PS-pellet had higher ACLY, YWHAZ, PPIA and FABP4 gene expression (p < .05) when compared to ALM-pellet control, with RUP by PS-pellet interactions (p < .01). Goats receiving additional RUP supplementation had increased (p < .05) STAT2 gene expression, whereas goats receiving PS-pellet supplementation showed increased STAT2 (p < .05) and a tendency to increase IRF3 (p = .07) gene expressions. In conclusion, the addition of PS-pellet or RUP supplementation has the potential to improve ADG and altered selected lipogenic and immune-related gene expressions., (© 2019 Blackwell Verlag GmbH.)

Background: Ten healthy adult Huang-huai white goats were selected and sacrificed by jugular vein bleeding after anaesthesia to observe the distribution characteristics of the histological structure of the intestinal mucosa, goblet cells, IgA, and CD3+ lymphocytes., Materials and Methods: Three sections of the duodenum, the jejunum, and the ileum were immediately collected and fixed with 4% paraformaldehyde for 72 h to prepare tissue sections. After haematoxylin and eosin, periodic acid Schiff, and immunohistochemical staining was performed, the distribution characteristics of goblet cells, IgA-positive cells, and CD3+ lymphocytes were observed. Results showed high columnar epithelial cells in the duodenum and jejunum of Huang-huai white goat and low columnar epithelial cells in the ileum mucosa., Results: Mucopolysaccharides secreted by intestinal goblet cells were mainly neutral, and the number of ileum goblet cells was significantly higher than that of the duodenum and the jejunum (p < 0.05). IgA-positive cells were distributed in the lamina propria of the duodenum, and the number of cells was significantly higher than that in the jejunum and the ileum (p < 0.01). The significant difference was found between the jejunum and the ileum (p < 0.01). The CD3+ cells in the intestinal mucosa were distributed in the lamina propria mucosae, and some of the positive cells in the jejunum were distributed between epithelial cells. CD3+ cells had the largest number in the jejunal lamina propria but had the lowest number in the ileum., Conclusions: The jejunum was significantly higher than the duodenum (p < 0.05), and the ileum was much less than the jejunum (p < 0.01).

The aim of the study was to investigate the use of serine protease from Yarrowia lipolytica yeast for reduction of milk proteins allergenicity. Whey protein concentrate (WPC-80), αs-casein and their hydrolysates were analyzed for the capacity to bind IgE and IgG antibodies present in sera from patients with cow milk protein allergy using a competitive ELISA. The hydrolysis of αs-casein and whey protein concentrate contributed to a significant reduction of their immunoreactive epitopes. In case of IgE antibodies, the lowest binding capacity was detected in the 24 h hydrolysates of both proteins in which the inhibition of the reaction was ≤20 and ≤68% for αs-casein and whey protein concentrate respectively. One hour hydrolysis of WPC-80 reduced the protein antigenicity, while the longer time (5 h) might lead to the exposure of new IgE - reactive epitopes., (Copyright © 2019. Published by Elsevier Ltd.)

This study aimed to determine if intrauterine-infused lipopolysaccharides (LPS) can be translocated to the mammary glands and induce an inflammatory response. Thirty-seven goats were divided into two experiments. Nineteen goats (control group, n = 9; LPS group, n = 10) were subjected to intravenous injection of LPS, and eighteen goats (control group, n = 8; LPS group, n = 10) were subjected to intrauterine infusion of LPS. Milk and blood samples were collected before and after the LPS challenge, to measure the blood leukocyte count (BLC), plasma LPS-binding protein (LBP), milk yield, milk somatic cell count (SCC), lactoferrin (LF), milk lactoperoxidase (LPO) activity, and pro- and anti-inflammatory cytokines in plasma and milk. Mammary gland tissues were collected from the parenchyma before and after the LPS challenge, for immunohistochemistry of LPS. In the intravenous injection experiment, the BLC (P < 0.001) and milk yield (P = 0.009) were lower, whereas the LF concentration (P < 0.001) and milk LPO activity (P < 0.001) were higher in the LPS group compared to that in the control group. LPS was detected in the mammary gland 3 and 24 h after intravenous injection of LPS. In the intrauterine infusion experiment, the mean concentrations of IL-1β and IL-6 in milk were higher in the LPS group compared to that in the control group (P = 0.004 and P = 0.017, respectively), whereas there were no changes in milk yield or SCC. LPS was detected in the connective tissues and interepithelial spaces of the alveoli of the mammary glands 24 h after intrauterine infusion of LPS. We conclude that intrauterine-infused LPS can be translocated to the mammary glands from the uterus, however, the amount of translocated LPS might not be enough to induce symptoms of clinical or subclinical mastitis., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Peste des petits ruminants is a highly contagious acute or subacute disease of small ruminants caused by the peste des petits ruminants virus (PPRV), and it is responsible for significant economic losses in animal husbandry. Vaccination represents the most effective means of controlling this disease, with virus-like particle (VLP) vaccines offering promising vaccine candidates. In this study, a PPRV VLP-based vaccine was developed using a baculovirus expression system, allowing for the simultaneous expression of the PPRV matrix (M), hemagglutinin (H), fusion (F) and nucleocapsid (N) proteins in insect cells. Immunization of mice and goats with PPRV VLPs elicited a robust neutralization response and a potent cellular immune response. Mouse studies demonstrated that VLPs induced a more robust IFN-γ response in CD4 + and CD8 + T cells than PPRV Nigeria 75/1 and recruited and/or activated more B cells and dendritic cells in inguinal lymph nodes. In addition, PPRV VLPs induced a strong Th1 class response in mice, as indicated by a high IgG2a to IgG1 ratio. Goat studies demonstrated that PPRV VLPs can induce the production of antibodies specific for F and H proteins and can also stimulate the production of virus neutralizing antibodies to the same magnitude as the PPRV Nigeria 75/1 vaccine. Higher amounts of IFN-γ in VLP-immunized animal serum suggested that VLPs also elicited a cellular immune response in goats. These results demonstrated that VLPs elicit a potent immune response against PPRV infection in small ruminants, making PPRV VLPs a potential candidate for PPRV vaccine development., Competing Interests: The authors declare no conflict of interest.

The leukocyte receptor complex (LRC) encodes a large number of immunoglobulin (Ig)-like receptors involved in the immune response, particularly in modulating natural killer (NK) cell function. The killer cell Ig-like receptors ( KIR ), the leukocyte Ig-like receptors ( LILR ), and a recently described novel Ig-like receptor family are highly variable between species, which is consistent with rapid evolution driven by selection pressure from pathogens. Among the species studied to date, only simians (such as humans) and bovids (such as cattle and goats) have an expanded complement of KIR genes and represent an interesting model to study KIR evolution. Using recently improved genome assemblies and an assembly of bacterial artificial chromosomes, we describe the structure of the LRC, and the KIR region in particular, in goats and compare this to sheep as the assemblies allow. These species diverged from a common ancestor ~10 million years ago and from cattle ~25 million years ago. We identified conserved KIR genes common to both goats and sheep and confirm a partial sheep haplotype shared between the Rambouillet and Texel breeds. Goats and sheep have independently expanded two novel KIR subgroups, and unlike cattle or any other mammal, they do not appear to possess a functional 3DL-lineage KIR gene. Investigation of LRC gene expression using available transcriptomic data for various sheep and goat tissues largely confirmed putative gene annotation and revealed that a relatively conserved caprinae -specific KIR subgroup is expressed in macrophages. The LILR and novel Ig-like receptors were also highly expressed across a diverse range of tissues. This further step toward our understanding of the LRC receptor repertoire will help inform future studies investigating immune response variation in these species., (Copyright © 2019 Schwartz, Sanderson, Bickhart, Smith and Hammond.)