Microproteomic-Based Analysis of the Goat Milk Protein Synthesis Network and Casein Production Evaluation.

Goat milk has been consumed by humans since ancient times and is highly nutritious. Its quality is mainly determined by its casein content. Milk protein synthesis is controlled by a complex network with many signal pathways. Therefore, the aim of our study is to clearly depict the signal pathways involved in milk protein synthesis in goat mammary epithelial cells (GMECs) using state-of-the-art microproteomic techniques and to identify the key genes involved in the signal pathway. The microproteomic analysis identified more than 2253 proteins, with 323 pathways annotated from the identified proteins. Knockdown of IRS1 expression significantly influenced goat casein composition (α, ß, and κ); therefore, this study also examined the insulin receptor substrate 1 (IRS1) gene more closely. A total of 12 differential expression proteins (DEPs) were characterized as upregulated or downregulated in the IRS1-silenced sample compared to the negative control. The enrichment and signal pathways of these DEPs in GMECs were identified using GO annotation and KEGG, as well as KOG analysis. Our findings expand our understanding of the functional genes involved in milk protein synthesis in goats, paving the way for new approaches for modifying casein content for the dairy goat industry and milk product development.

The Expression of Selected Cytokine Genes in the Livers of Young Castrated Bucks after Supplementation with a Mixture of Dry Curcuma longa and Rosmarinus officinalis Extracts.

The study aims to determine the effect of supplementation with a mixture of Curcuma longa and Rosmarinus officinalis extracts (896:19 ratio) on the expression of 15 cytokine genes in the livers of 20 castrated goat bucks. Two equal groups were created: treated and control groups. The treated group was provided a mixture (1.6 g/day/buck) for 124 days. Liver tissue samples were collected after slaughter. The gene expression was analyzed using RT-qPCR with two reference genes. Variance analysis was conducted using a model with the group fixed effect. IL-2 and IL-8 expression was below the detection level. No differences were found for IL-1α, IL-1ß, IL-4, IL-6, IL-10, IL-16, IFN-α, IFN-ß, TNF-α, and CCL4 expressions, suggesting that supplementation does not activate cytokine production in the healthy hepatocytes. The treated group demonstrated lower IL-12 expression (p < 0.05) and a tendency for higher IL-18 and INF-γ (0.05 < p < 0.10) expressions, which may indicate a hypersensitivity resulting from excessive supplement dose. The increased IFN-γ expression could be caused by the increased IL-18 expression. If a small dose of extract can induce an allergic reaction in young goat bucks, it is also possible that humans may be susceptible to an overdose of curcumin and/or turmeric extracts.

Effect of Supplementation with Curcuma longa and Rosmarinus officinalis Extract Mixture on Acute Phase Protein, Cathelicidin, Defensin and Cytolytic Protein Gene Expression in the Livers of Young Castrated Polish White Improved Bucks.

Goats are an excellent animal model for research on some physiological and pathophysiological processes in humans. The search for supplements that prevent homeostasis disorders and strengthen the immune system is necessary to reduce the risk of many diseases in both humans and animals. The aim of the study was to analyze the effect of supplementation with a mixture of dried extracts of Curcuma longa and Rosmarinus officinalis on the expression of acute-phase protein (SAA, HP, CRP, LALBA, AGP, CP, FGA, FGB, and FGG), cathelicidin (BAC5, BAC7.5, BAC3.4, MAP28, MAP34, and HEPC), beta-defensin-1 (GBD1, DEFB1), and beta-defensin-2, and cytolytic protein (LIZ and LF) genes in the livers of young castrated bucks of the Polish White Improved breed. The higher expression of LF in the control group suggests that it is important for the first line of hepatic immune defense and its expression is downregulated by the mixture of turmeric and rosemary extracts; thus, the spice-herb mixture mutes its activity. The lower expression of FGB and the higher expression of BAC5 genes in the livers of healthy, young castrated bucks who were administered the supplement suggest the silencing effects of the mixture on the acute-phase response and the stimulating effect on the antimicrobial activity of the immune system.

A large-scale study on the seroprevalence of small ruminant lentiviral infection in the Polish goat population.

A large-scale study was carried out in a Polish goat population in 2014-2022 to determine the herd-level (between-herd) and within-herd seroprevalence of small ruminant lentivirus (SRLV) infection. A total of 8354 adult goats (aged >1 year) from 165 herds located in various regions of Poland were serologically tested using a commercial ELISA. One hundred twenty eight herds were randomly selected while 37 were enrolled based on convenience non-random sampling. At least 1 seropositive result was obtained in 103 / 165 herds. For all these herds the probability that they were truly positive (herd-level positive predictive value) was calculated. It was ≥ 90% in 91 seropositive herds and 73% to < 90% in 12 herds in which only 1-4 goats were seropositive (22 goats in total). The seropositive goats in the latter herds were retested using a different commercial ELISA and 14 goats (9 males and 5 females) from 9 herds were confirmed to be seropositive (serial testing). The true herd-level seroprevalence was estimated at 61% (95% confidence interval [CI 95%]: 53%-68%). It differed significantly between herd size classes (p = 0.003): the highest prevalences were found in the medium (51 - 100 adult goats) and large herds (>100 adult goats) - 72% (CI 95%: 56-84%) and 86% (CI 95%: 67%-95%), respectively, while prevalences in very small (≤ 20 adult goats) and small herds (21 - 50 adult goats) were 46% (CI 95%: 34%-59%) and 57% (CI 95%: 43%-70%), respectively. The true herd-level seroprevalence differed significantly also between geographical regions of Poland (p = 0.003), with the highest values in the north-western and the lowest in the southern region of the country. The true within-herd seroprevalence estimated using a Bayesian approach ranged from 0.7% to 100% with the median (IQR) of 42% (17%-84%), and did not vary significantly between herd size classes (p = 0.393) or geographical regions of Poland (p = 0.570). Concluding, SRLV infection is widespread in the Polish goat population, the north-western region of Poland is most extensively infected, and herds counting > 50 adult goats are more often infected.

Health potential of fermented goat dairy products: composition comparison with fermented cow milk, probiotics selection, health benefits and mechanisms.

Goat milk as a preferable probiotic vehicle has been investigated and the contribution of fermented goat dairy products to the nutritional and economic wellbeing of the world is tremendous. This review presents the recent progress on fermented goat dairy products, including probiotic selection, composition comparison to fermented cow milk, health effects, and related mechanisms. Fermented goat milk maintains a better nutritional profile in comparison to fermented cow milk with higher values of protein, minerals (Ca, Mg, Fe, Cu, Zn and Se), vitamins (A, D3 and B12) and some fatty acids. Lactobacillus is the predominant genus used in goat milk fermentation and endows goat milk with higher functional value, including gut microbiota regulation, anti-microbial and anti-inflammatory functions, hypocholesterolemic effects, antioxidant effects, hypotensive effects, bone health, anemia recovery, anti-obesity, and anti-atherogenic function. The corresponding mechanisms have been elucidated at the molecular level. A series of collection on probiotics starters, fermentation strategy and characteristics of fermented goat dairy products are performed. Although the industrial applications of fermented goat milk remain underdeveloped, the improved functional annotation and fermentation strategy identified in this review provide a bright future and an excellent framework for the future fermented goat dairy market.

Serological testing of an equal-volume milk sample - a new method to estimate the seroprevalence of small ruminant lentivirus infection?

BACKGROUND: In cattle attempts to evaluate within-herd prevalence of various infectious and parasitic diseases by bulk-tank milk (BTM) testing with ELISA have been made with moderate success. The fact that BTM is composed of variable and unknown volumes of milk from individual lactating animals weakens the relationship between numerical result of the ELISA and the within-herd prevalence. We carried out a laboratory experimental study to evaluate if a pooled milk sample created by mixing an equal volume of individual milk samples from seropositive and seronegative goats, henceforth referred to as an equal-volume milk sample (EVMS), would allow for accurate estimation of within-herd seroprevalence of caprine arthritis-encephalitis (CAE) using 3 different commercial ELISAs. By mixing randomly selected milk samples from seronegative and seropositive goats, 193 EVMS were created - 93 made of seronegative samples and 100 with the proportion of seropositive individual milk samples (EVMS%POS) ranging from 1 to 100%. EVMS%POS could be considered as a proxy for the within-herd seroprevalence. Then, OD of EVMS (ODEVMS) of the 193 EVMS was measured using 3 commercial ELISAs for CAE - 2 indirect and 1 competitive. RESULTS: The cut-off values of ODEVMS indicating SRLV infection were determined. The regression functions were developed to link ODEVMS with EVMS%POS. A significant monotonic relationship between ODEVMS measured with 2 commercial indirect ELISAs and EVMS%POS was identified. Two regression models developed on this basis described approximately 90% of variability and allowed to estimate EVMS%POS, when it was below 50%. High ODEVMS indicated EVMS%POS of > 50%. CONCLUSION: Our study introduces the concept of serological testing of EVMS as a method of detecting SRLV-infected herds and estimating the proportion of strongly seropositive goats. Further field studies are warranted to assess practical benefits of EVMS serological testing.

Single nucleotide polymorphisms in the bovine SLC2A12 and SLC5A1 glucose transporter genes - the effect on gene expression and milk traits of Holstein Friesian cows.

In this study, novel single nucleotide polymorphisms (SNPs) were found in the 5'-regulatory regions (promoters) of the bovine glucose transporter (GT) genes SLC2A12 and SLC5A1. These polymorphisms were shown to associate with certain milk production traits in HF cows, including milk yield, milk composition, and somatic cell count. It was shown that the SNP g.-671C > G (NC_037336.1: g.72224078C > G) in the SLC2A12 gene could be an effective marker of cattle production traits and that genotypes CC and CG are associated with the best productivity. The polymorphisms found in the SLC5A1 gene promoter also influenced milk production traits in HF cows, albeit to a lesser extent, and we propose that these polymorphisms could be useful as genetic markers for milk production traits in marker-assisted selection (MAS); however, this must be confirmed on larger populations of cattle. In addition, the presence of polymorphisms within promoter regions appears to affect the expression of GT genes in the cow mammary gland and modify transcription factor (TF) binding capacity.

Effect of Supplementation with Organic Selenium or Turmeric and Rosemary Mixture on Beta-Defensin Content in Goat Milk.

The present study examines the effects of diet supplementation with an organic selenium or herb-spice mixture on beta-defensin1 (GBD-1) and beta-defensin2 (GBD-2) concentrations in goat milk. Herd-I, consisting of Polish White (PWI) and Fawn Improved (PFI) goats, received supplementation with organic or inorganic selenium (controls). All goats were free from parasites, mastitis, and small ruminant lentivirus infection. Herd-II, consisting of PWI goats, either received a turmeric-rosemary mixture (experimental) or not (controls). The Herd I control group demonstrated higher fat, free fatty acid, and somatic cell levels and lower protein and lactose contents than Herd II controls. The GBD-1 concentration was below the detection limit in both herds. Herd I controls demonstrated higher GBD-2 concentrations in milk than Herd II controls. In addition, lower GBD-2 concentrations were noted in PWI goat milk. Organic selenium or rosemary-turmeric mixture supplementation had no effect on the GBD-2 content in the milk of healthy goats. The higher GBD-2 concentration observed in Herd Ic than in Herd IIc may suggest that the type of basal diet affects defensin secretion.

miRNA expression patterns in blood leukocytes and milk somatic cells of goats infected with small ruminant lentivirus (SRLV).

The study aims to determine the selected miRNAs expression in milk somatic cells (MSC) and blood leukocytes (BL) of SRLV-seronegative (SRLV-SN) and SRLV-seropositive (SRLV-SP) goats. A functional in silico analysis of their target genes was also conducted. MiR-93-5p and miR-30e-5p were expressed only in BL, while miR-144 was expressed only in MSC, regardless of SRLV infection. In the SRLV-SP goats, higher miR-214-3p and miR-221-5p levels were found in the MSC than in the BL. Only miR-30e-5p was influenced by the lactation stage in BL in both groups, while only miR-93-5p was altered in BL of SRLV-SN goats. The target gene protein products exhibited contradictory functions, protecting the host from virus on the one hand and assisting viruses in their life cycle on the other. The differential expression of the miRNAs observed between the MSC and BL of SRLV-SP goats may suggest that the local immune response to the infection in the udder differs from the systemic response, and acts independently. Some miRNAs demonstrated different expression between lactation stages. It may be influenced by the metabolic burden occurring in early lactation and its peak. Some of the studied miRNAs may influence viral infection by regulating the expression of their target genes.

The Prevalence of Histopathological Features of Pneumonia in Goats with Symptomatic Caprine Arthritis-Encephalitis.

Chronic interstitial pneumonia (CIP) is a main pathology of sheep infected with small ruminant lentivirus (SRLV). Caprine arthritis-encephalitis (CAE) is caused by the same pathogen; however, the presence of CIP has been only occasionally reported in SRLV-infected goats. We carried out a cross-sectional study to determine the prevalence of histopathological lesions indicative of CIP in goats with symptomatic CAE, and to investigate whether CIP was associated with a higher prevalence of other types of pneumonia (purulent bronchopneumonia, fibrinous pleuropneumonia) or bacterial infections. Lung specimens and bronchial swabs were collected for histopathological and bacteriological examination, respectively, from 116 goats from a CAE-affected herd. All goats were euthanized due to severe clinical signs of CAE. The goats were seropositive for SRLV infection in two different ELISAs and the presence of SRLV antigen in the lung tissue was confirmed by immunohistochemistry. Histopathologically, pneumonia of any type was confirmed in 82 goats (70.7%) and CIP was present in 67 goats (57.8%). In most goats, the severity of the histopathological features of pneumonia was mild. Bacteria were detected in bronchial swabs from 73 goats (62.9%). CIP proved to be significantly positively linked to the occurrence of purulent bronchopneumonia (p < 0.001), fibrinous pleuropneumonia (p = 0.001), and of the infection of lungs with bacteria capable of causing pneumonia (p = 0.050). The causal character of these associations should be considered and warrants further investigation.

Lack of Fetal Protection against Bovine Viral Diarrhea Virus in a Vaccinated Heifer.

The aim of the report was to present the circulation of BVDV (bovine viral diarrhea virus) in the cattle population and determine the cause of the failure of vaccination failure leading to the birth of the PI (persistently infected) calf. The case study was carried out at the BVDV-free animal breeding center and cattle farm, where the vaccination program against BVDV was implemented in 2012, and each newly introduced animal was serologically and virologically tested for BVDV. In this case, a blood sample was taken from a 9-month-old breeding bull. Positive RT-PCR and negative ELISA serology results were obtained. The tests were repeated at 2-week intervals, and the results confirmed the presence of the virus and the absence of specific antibodies, i.e., persistent infection. Additionally, sequencing and phylogenetic analysis were performed, and the BVDV-1d subgenotype was detected. The results of this study showed that pregnant heifers and cows that are vaccinated multiple times with the killed vaccine containing BVDV-1a may not be fully protected against infection with other subgenotypes of BVDV, including their fetuses, which can become PI calves.

Longitudinal Study on Seroreactivity of Goats Exposed to Colostrum and Milk of Small Ruminant Lentivirus-infected Dams.

Introduction: Small ruminant lentivirus (SRLV) causes caprine arthritis-encephalitis in goats and maedi-visna disease in sheep. Transmission is via ingestion of colostrum and milk from infected dams or long-term direct contact between animals. Lifelong seroconversion can occur several weeks after infection via ingestion. However, sub-yearling lambs that ingest contaminated colostrum may be able to clear the infection and become seronegative. Whether a similar phenomenon occurs in goats remains unknown. Therefore, the serological status of goats was studied longitudinally from the moment of natural exposure to colostrum and milk of SRLV-positive dams through the age of 24 months. Material and Methods: Between February 2014 and March 2017 a dairy goat herd was studied which had been infected with SRLV for more than 20 years and carried maedi-visna virus-like genotype A subtype A17. Thirty-one kids born to dams seropositive for SRLV for at least a year beforehand were followed. They ingested colostrum immediately after birth and then remained with their dams for three weeks. The goats were tested serologically every month using two commercial ELISAs. The clinical condition of the goats was also regularly assessed. Results: Out of 31 goats, 13 (42%) seroconverted at the age ranging from 3 to 22 months with a median of 5 months. Two goats seroconverted in the second year of life. The other eleven did so before the age of one year; two of these reverted to seronegative status. Only 9 out of 31 goats (29%) seroconverted in the first year of life and remained seropositive. They were early and stable seroreactors to which SRLV was transmitted lactogenically. The age at which they seroconverted ranged from 3 to 10 months with a median of 5 months. In 8 of the 18 persistently seronegative goats, a single isolated positive result occurred. No goats showed any clinical signs of arthritis. The level of maternal antibodies at the age of one week did not differ significantly between the stable seroreactors and the remainder. Conclusion: Seroconversion appears to occur in less than 50% of goats exposed to heterologous SRLV genotype A via ingestion of colostrum and milk from infected dams and is delayed by 3-10 months. The natural lactogenic route of transmission of SRLV genotype A in goats appears to be less effective than this route of genotype B transmission reported in earlier studies.

Expression of cytokines in dairy cattle mammary gland parenchyma during chronic staphylococcal infection.

The study aim was to determine the expression of genes potentially related to chronic mastitis at the mRNA and protein levels, viz. chemokine C-C motif receptor 1 (CCR1), C-C motif chemokine ligand 2 (CCL2), C-C motif chemokine ligand 5 (CXCL5), tumor necrosis factor α (TNFα), interleukin 1ß (IL-1ß), interleukin 6 (IL-6), interleukin 8 (IL-8), interleukin 18 (IL-18), in bovine mammary gland parenchyma. The study examines the differences in expression of selected genes between cows with chronic mastitis caused by coagulase-positive (CoPS) or coagulase-negative staphylococci (CoNS) and those with healthy udders (H). Samples were collected from the udder quarters from 40 Polish Holstein-Friesian cows; 54 of these samples were chosen for analysis based on microbiological analysis of milk taken two days before slaughter. They were categorized into three groups: CoPS (N = 27), CoNS (N = 14) and H (N = 13). The RNA expression was analyzed by RT-qPCR and protein concentration by ELISA. No differences in the mRNA levels of seven genes (TNFα, IL-18, CCR1, IL-1ß, CCL2, IL-8, IL-6) and four proteins (TNFα, IL-18, CCR1, IL-1ß) were identified between the CoPS and H groups. Higher transcript levels of CXCL5 (p ≤ 0.05) gene were noted in CoPS than in H. Compared to H, higher concentrations of IL-8 and CXCL5 (p ≤ 0.05) were observed in CoPS (0.05 < p < 0.1) and CCL2 (0.05 < p < 0.1) in CoNS, while lower levels of Il-6 were found in CoPS. This may suggest that during chronic mastitis the organism stops producing pro-inflammatory cytokines, probably to protect the host tissues against their damage during prolonged infection.

A Comparison of Oxidative Stress Biomarkers in the Serum of Healthy Polish Dairy Goats with Those Naturally Infected with Small Ruminant Lentivirus in the Course of Lactation.

The present study examines the effects of natural infection by small ruminant lentivirus (SRLV) in the two most common goat breeds in Poland, i.e., Polish white improved and polish fawn improved. It focuses on biomarkers of oxidative stress, oxidatively modified proteins and antioxidant defenses, ceruloplasmin level as an acute phase protein, and the activities of antioxidant enzymes in the goat serum. It was conducted on 24 goats divided into two equal groups: one SRLV-seropositive (SRLV-SP) and another SRLV-seronegative (SRLV-SN). Both groups were identical in terms of breed and parity. Despite infection, the SRLV-SP goats demonstrated no symptoms of caprine arthritis-encephalitis. In addition, the SRLV-SP goats did not reveal pronounced dysfunctions in oxidative stress biomarkers in the serum compared to the SRLV-SN animals. However, both groups demonstrated elevated levels of the aldehydic and ketonic derivatives of oxidatively modified proteins during the lactation period. In addition, both groups retained a high total antioxidant capacity in serum despite the decrease of enzyme antioxidant defenses throughout the 200-day lactation period.

Does Small Ruminant Lentivirus Infection in Goats Predispose to Bacterial Infection of the Mammary Gland? A Preliminary Study.

The aim of this study was to determine whether asymptomatic small ruminant lentivirus seropositive (SRLV-SP) goats were more susceptible to bacterial infection of the udder when lactating by comparing the presence and species of pathogenic bacteria in their milk with the values for seronegative goats (SRLV-SN). Milk samples were collected during morning milking on days 20, 40, 60, 150, and 210 of lactation for three consecutive years and subjected to bacteriological examination. Staphylococcus caprae and S. xylosus were the most frequent strains identified in both SRLV-SP and SRLV-SN goats. The prevalence of pathogenic bacteria was the highest in the 1st lactation, regardless of SRLV status. Moreover, the prevalence of pathogenic bacteria was significantly higher in SRLV-SP goats, but only those in the 5th or further lactation (p = 0.010). This suggests a relationship between long-lasting SRLV infection and susceptibility to bacterial infections of the udder.

The effect of single-nucleotide polymorphism in the promoter region of bovine alpha-lactalbumin (LALBA) gene on LALBA expression in milk cells and milk traits of cows.

Polymorphisms of milk protein genes have been proposed as candidate markers for dairy production traits in cattle. In the present study, a polymorphism was detected in the 5'-flanking (promoter) region of the bovine alpha-lactalbumin (LALBA) gene, a T/C transition located at nucleotide -1,001 relative to the transcription start site g.-1001T > C (NC_037332.1:g.31183170T > C), which is recognizable with PstI restriction endonuclease. In silico analyses showed that this mutation created novel retinoid X receptor alpha and vitamin D receptor transcription factor binding sites. Real-time PCR found that cows with different genetic variants of the promoter demonstrated different levels of expression of LALBA mRNA in milk somatic cells (MSCs). The TT genotype cows demonstrated low expression, whereas those with CT demonstrated much higher expression (P < 0.05). ELISA analysis found milk LALBA protein levels also differed between the TT and CT cows (P < 0.05) and that these levels were not correlated with the mRNA abundance in MSC. Association analysis found that the g.-1001T > C polymorphism in the promoter region of the LALBA gene influenced milk production traits in Polish Holstein-Friesian cows. High daily milk yield and dry matter yield, and high lactose yield and concentration were associated with the TT genotype. The TT genotype cows also had a lower number of somatic cells in the milk, considered as an indicator of udder health status. Therefore, the TT genotype could be more desirable from the breeder's perspective.

Diagnostic accuracy of three commercial immunoenzymatic assays for small ruminant lentivirus infection in goats performed on individual milk samples.

Caprine arthritis-encephalitis (CAE) caused by small ruminant lentivirus (SRLV) infection is one of the most widespread and devastating diseases of goats. Serological methods, mainly immunoenzymatic assays (ELISA), are the mainstay of CAE diagnostics. Even though blood is still the most commonly tested material, animal welfare issues and increasing costs of veterinary service prompt the development of serological methods based on milk testing. Several different types of ELISAs for CAE are available on the market. All of them perform well on serum, however their diagnostic accuracy for testing milk has not been so far compared. Therefore, we carried out the study in 5 dairy goat herds in Poland whose previous epidemiological situation regarding CAE was known. Paired serum and milk samples were collected from all adult females (n = 420) and tested with 3 commercial ELISAs - indirect ELISA based on the whole-virus antigen (wELISA), indirect ELISA based on the recombined transmembrane and capsid protein (TM/CA-ELISA), and competitive ELISA based on the surface glycoprotein (SU-ELISA). Milk was tested as lactoserum at dilution of 1/2 in wELISA and TM/CA-ELISA, and undiluted in SU-ELISA. The true status of goats was based on the composite reference standard comprising the results of all three ELISAs done on serum and the true prevalence of SRLV infection in the herd of origin. 243 (57.9 %) goats were classified as truly positive and 177 (42.1 %) goats as truly negative. Diagnostic accuracy was evaluated using the area under the ROC curve (AUROC) as well as sensitivity (Se) and specificity (Sp) for a range of cut-off values. AUROC was 98.8 % (CI 95 %: 97.5 %, 100 %) for wELISA, 97.9 % (CI 95 %: 96.5 %, 99.2 %) for TM/CA-ELISA, and 91.7 % (CI 95 %: 88.9 %, 94.5 %) for SU-ELISA. At the cut-off values recommended by the manufacturers both indirect ELISAs were highly sensitive (89.3 % and 91.4 %, respectively) and highly specific (98.3 % and 95.5 %, respectively), whereas SU-ELISA had only moderate Se (71.2 %) at comparably high Sp (96.6 %). Nevertheless, the optimal cut-off values were lower than those recommended by manufacturers for serum - sample-to-positive control serum ratio (S/P%) of 10 % for wELISA, S/P% of 80 % for TM/CA-ELISA, and percentage inhibition of 23 % for SU-ELISA. Concluding, the study shows that wELISA and TM/CA-ELISA may be interchangeably used for testing individual goat milk samples for SRLV infection. Diagnostic sensitivity and specificity of these ELISAs appear not to be lower on milk than on serum. SU-ELISA is considerably less sensitive on milk samples than indirect ELISAs.

Gene Expression Profile in Peripheral Blood Nuclear Cells of Small Ruminant Lentivirus-Seropositive and Seronegative Dairy Goats in Their First Lactation.

The immune response to a viral antigen causes inflammatory cell infiltration to the tissue, which creates a suitable environment for the replication of the virus in macrophages, and the recruitment of more monocytes to the site of infection, or latently infected monocytes. The aim of the study was to analyze the transcriptomic profile of peripheral blood nuclear cells isolated from SRLV-seropositive and SRLV-negative goats at the peak of their first lactation. SRLV-seropositive goats were probably infected via colostrum. Custom transcriptomic microarrays for goats were designed and developed, namely the Capra hircus gene expression array, which features ~50,000 unique transcripts per microarray. Only four genes were differentially expressed, with up-regulated expression of the GIMAP2, SSC5D and SETX genes, and down-regulated expression of the GPR37 gene in SRLV-seropositive vs. SRLV-seronegative goats. However, in an RT-qPCR analysis, the result for the SETX gene was not confirmed. The differences in the expressions of the studied genes indicate an active inflammatory process in the SRLV-seropositive goats at the early stage of infection.

MicroRNA expression profile in bovine mammary gland parenchyma infected by coagulase-positive or coagulase-negative staphylococci.

MicroRNAs (miRNAs) are short, non-coding RNAs, 21-23 nucleotides in length which are known to regulate biological processes that greatly impact immune system activity. The aim of the study was to compare the miRNA expression in non-infected (H) mammary gland parenchyma samples with that of glands infected with coagulase-positive staphylococci (CoPS) or coagulase-negative staphylococci (CoNS) using next-generation sequencing. The miRNA profile of the parenchyma was found to change during mastitis, with its profile depending on the type of pathogen. Comparing the CoPS and H groups, 256 known and 260 potentially new miRNAs were identified, including 32 that were differentially expressed (p ≤ 0.05), of which 27 were upregulated and 5 downregulated. Comparing the CoNS and H groups, 242 known and 171 new unique miRNAs were identified: 10 were upregulated (p ≤ 0.05), and 2 downregulated (p ≤ 0.05). In addition, comparing CoPS with H and CoNS with H, 5 Kyoto Encyclopedia of Genes and Genomes pathways were identified; in both comparisons, differentially-expressed miRNAs were associated with the bacterial invasion of epithelial cells and focal adhesion pathways. Four gene ontology terms were identified in each comparison, with 2 being common to both immune system processes and signal transduction. Our results indicate that miRNAs, especially miR-99 and miR-182, play an essential role in the epigenetic regulation of a range of cellular processes, including immunological systems bacterial growth in dendritic cells and disease pathogenesis (miR-99), DNA repair and tumor progression (miR-182).

Transcripts and protein levels of CSN1S1 and CSN3 genes in dairy cattle mammary gland secretory tissue during chronic staphylococcal infection.

Our objective was to determine the influence of chronic coagulase-positive staphylococci (CoPS) or coagulase-negative staphylococci (CoNS) infection on the mRNA and protein levels of two main milk proteins responsible for cheese curd quantity and quality, alpha-S1-casein (CSN1S1) and kappa-casein (CSN3). Measurements were made in cow mammary parenchyma with a prevalence of secretory tissue (MGST). Samples of MGST were collected from the separate quarters and divided into CoPS, CoNS and bacteria-free (H) groups according to the microbiological status of the quarter milk. No differences in CSN1S1 and CSN3 mRNA level were found between groups, however, CSN1S1 protein level was significantly higher in the H group than the CoNS group, and CSN3 protein level was significantly higher in H than CoPS group. Hence, while the CSN1S1 and CSN3 genes appear to be constitutively expressed at the mRNA level in dairy cow MGST during mastitis, CoNS infection negatively affected CSN1S1 protein level, and CoPS infection negatively affected CSN3 protein level. The lack of change at the mRNA level suggests that staphylococcal infection may affect the post-transcriptional or post-translational modifications.