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.

Application of a complete blood count to screening lethargic and anorectic cats for pancreatitis.

BACKGROUND: Feline pancreatitis (FP) is an important health problem of cats. Its diagnostics is based on the combination of quantification of serum pancreatic lipase immunoreactivity (fPLI) and abdominal ultrasonography (AUS). These modalities allow for establishing highly specific diagnosis, however they are relatively expensive and time-consuming. On the other hand, a screening test of high sensitivity which would allow to rule out FP on the first visit without a considerable increase of costs would be clinically useful. To evaluate accuracy of nonspecific inflammatory biomarkers based on complete blood count (CBC) in diagnosing FP 73 client-owned cats with signs of lethargy and reduced appetite lasting for at least 2 days before presentation were enrolled in the cross-sectional study. They were examined with fPLI assay and AUS and classified as cats with very low risk of FP when fPLI ≤3.5 µg/L and AUS negative for FP, or as cats with increased risk of FP in the case of any other combination of results. Then, 7 various CBC measurements were measured in each cat and linked to the risk of FP using the multivariable logistic regression. RESULTS: Five CBC measurements turned out to be significantly associated with the risk of FP - total leukocyte count (WBC; crude odds ratio(ORcrude) = 12.2; CI 95%: 1.52, 98.5), total neutrophil count (ORcrude = 5.84; CI 95%: 1.22, 27.9), band neutrophil count (BNC; ORcrude = 6.67; CI 95%: 1.98, 22.4), neutrophil-to-lymphocyte ratio (ORcrude = 3.68; CI 95%: 1.25, 10.9), and eosinophil count (EC; ORcrude = 0.34; CI 95%: 0.12, 0.96). The model based on WBC, BNC, and EC proved to have at least fair diagnostic potential (area under ROC curve 82.7%; CI 95%: 72.8%, 92.5%). When WBC <  18 G/L, BNC <  0.27 G/L, and EC >  0.3 G/L was considered as a negative result, and any other combination as the positive result, the CBC model had high sensitivity (91.8%; CI 95%: 80.8%, 96.8%) at a relatively low specificity (58.3%; CI 95%: 38.8%, 75.5%). CONCLUSION: The combination of three CBC measurements is an immediately available and fairly accurate screening method for identification of lethargic and anorectic cats with increased risk of FP.

Prevalence of anthelmintic resistance of gastrointestinal nematodes in Polish goat herds assessed by the larval development test.

BACKGROUND: Helminthic infections, in particular those caused by gastrointestinal nematodes (GIN), are found worldwide and are among the most economically important diseases of goats. Anthelmintic resistance (AR) in GIN of goats is currently present worldwide, and single- or multidrug resistant species are widespread. The aim of this study was to determine the prevalence of AR to benzimidazoles (BZ), macrocyclic lactones (ML) and imidazothiazoles represented by levamisole (LEV) in the Polish goat herds by using an in vitro larval development test, which is useful especially in large-scale epidemiological surveys. RESULTS: This cross-sectional study was conducted from September 2018 to June 2019 and enrolled 42 dairy goat herds scattered over the entire country. The most commonly used anthelmintic class in goat herds in Poland were BZ (92%), followed by ML (85%) and LEV (13%). BZ-resistant GIN populations were found in 37 herds (88%, CI 95%: 75 to 95%), ML-resistant GIN populations in 40 herds (95%, CI 95, 84 to 99%), and LEV-resistant GIN populations in 5 herds (12%, CI 95%: 5 to 25%). Multidrug resistance involving all three anthelmintic classes was found in 5 herds (12%, CI 95, 5 to 25%). Based on the morphological features of stage 3 larvae the main resistant GIN turned out to be Haemonchus contortus and Trichostrongylus spp. The use of BZ and frequency of anthelmintic treatments were significantly related to the presence of AR to BZ in Polish goat herds. CONCLUSIONS: This cross-sectional study demonstrates the existence of AR to BZ, ML and LEV on Polish goat farms. Resistance to BZ and ML is widespread, while AR to LEV is currently at a low level. A considerable proportion of herds harbours multidrug resistant GIN, which requires further consideration. An effective anthelmintic treatment strategy, reasonable preventive measures and better understanding of the resistance-related management practices by farmers and veterinarians may delay further development of AR.

Profile of serum lipid metabolites of one-week-old goat kids depending on the type of rearing.

BACKGROUND: Weaning of goat kids immediately after birth and feeding them on bovine or heat-treated caprine colostrum, referred to as snatching, is considered an effective control measure in some infectious diseases. The study was carried out in one-week-old goat kids to gain insight into the profile of lipid metabolites and to investigate the influence of snatching on kids' metabolism. Fifty-two goat kids born to 23 female goats were included in the study - 22 kids were weaned immediately after birth and kept isolated from their mothers; 30 remaining kids were left with their mothers for next 3 weeks so that they could nurse on dams' milk at will. Blood was collected at the age of 1 week and serum was obtained by centrifugation. The concentration of lipid metabolites was determined with mass spectrometry using a commercial MxP® Quant 500 kit (Biocrates Life Sciences AG, Innsbruck, Austria). RESULTS: Concentration of 240 lipid metabolites belonging to 10 lipid classes was above the limit of detection of the assay. These lipid metabolites were quantified and included in the analysis. Concentration of 2 lipid classes (acyl-alkyl-phosphatidylcholines and ceramides) and 31 lipid metabolites (14 triacylglycerols, 5 acyl-alkyl-phosphatidylcholines, 2 diacylphosphatidylcholines, 1 lyso-phosphatidylcholine, 5 ceramides, 2 sphingomyelins, and 2 cholesterol esters) differed significantly between the two groups of kids. CONCLUSION: Snatching of kids results in reduction of serum concentration of lipid metabolites, however, the magnitude of this phenomenon does not seem to be sufficient to negatively affect kids' health condition. This study is the first in which the broad set of lipid metabolites of young ruminants was quantified using the novel metabolomic assay MxP® Quant 500 kit.

The first report of multidrug resistance in gastrointestinal nematodes in goat population in Poland.

BACKGROUND: Prophylactic anthelmintic treatment with one of three basic classes of anthelmintics (benzimidazoles, macrocyclic lactones and imidazothiazoles) is still the mainstay of control of gastrointestinal nematode infections in small ruminants worldwide. As a consequence, anthelmintic resistance is a serious threat to small ruminant health and production. While the resistance to one class of anthelmintics has already been reported in most of countries, the newly-emerging problem is the resistance to two or even all of classes referred to as multidrug resistance. This study aimed to evidence the presence of multidrug resistance of gastrointestinal nematodes in goats in Poland. RESULTS: The combination of one in vivo method (fecal egg count reduction test) and two in vitro methods (egg hatch test and larval development test) performed in two goat herds in the southern Poland showed the presence of gastrointestinal nematodes resistant to fenbendazole and ivermectin in both herds. Moreover, in one herd it revealed the development of resistance to the last effective anthelmintic, levamisole, in response to one-year intensive use. Haemonchus contortus was the most prevalent gastrointestinal nematode in samples in which resistance to benzimidazoles and ivermectin was found, whereas Trichostrongylus colubriformis predominated when resistance to levamisole was observed. CONCLUSION: This study shows for the first time that multidrug resistance of gastrointestinal nematodes to three basic classes of anthelmintics is already present in goat population in Poland. Moreover, it may indicate that different species or genera of gastrointestinal nematodes are responsible for the resistance to specific anthelmintics.

The expression of cytokines in the milk somatic cells, blood leukocytes and serum of goats infected with small ruminant lentivirus.

BACKGROUND: The present study aimed to determine the expression of cytokines, which is associated with the immunological response of dairy goats against small ruminant lentivirus (SRLV). The study was conducted on 26 dairy goats in their second to sixth lactation, which were divided by breed and parity into two groups: SRLV naturally infected (N = 13) and non-infected (N = 13) animals. All goats in the study were asymptomatic. The milk and blood samples, which served as studied material were taken on days 7, 30, 120 and 240 of the lactation. The gene and protein expression of several cytokines was studied using Real-Time PCR and ELISA methods. RESULTS: INF-ß and INF-γ expression was down-regulated in the milk somatic cells (MSC) of SRLV-infected goats. However, an increased concentration of INF-ß was observed in the MSC in SRLV-infected goats, while INF-γ expression was not observed in both SRLV-infected and non-infected animals The SRLV-infected goats also displayed decreased expression of IL-1α, IL-1ß, IL-6 and INF-γ genes in the blood leukocytes,with IL-1α, IL-1ß and IL-6 protein levels also being decreased in the sera. TNF-α was the only gene that demonstrated increased expression in both the MSC and the blood of infected animals; however, no such overexpression was observed at the protein level. CONCLUSIONS: SRLV probably influences the immune system of infected animals by deregulating of the expression of cytokines. Further, epigenetic studies may clarify the mechanisms by which SRLV regulates the gene and protein expression of the host.

The impact of organic vs. inorganic selenium on dairy goat productivity and expression of selected genes in milk somatic cells.

The aim of this study was to determine the effect of diet supplemented with selenized yeast (Se-yeast) on milk yield and milk composition of goats and expression of casein and mammary-gland-immune system genes in milk somatic cells (MSC). Twenty-four dairy goats in their second to fourth lactations were divided into control and experimental groups, balanced according to lactation number and breed (Polish White or Fawn Improved). Morning milk and blood samples were collected four times during lactation (on the 21st, 70th, 120th, 180th day after kidding). The control and experimental groups were fed diets with 0.7 mg inorganic Se/goat/day (sodium selenite) or 0.6 mg organic Se/goat/day (selenized yeast), respectively. Milk, fat and protein yields during lactation as well as average somatic cell count, fat, protein and lactose contents in milk were evaluated. Microelements in milk and blood serum and biochemical parameters in blood serum were determined at the beginning and the end of the experiment. The expression levels of the genes encoding αS1-casein (CSN1S1), αS2-casein (CSN1S2), κ-casein (CSN3), interleukin 8 (IL-8), serum amyloid A3 (SAA3), interleukin 1ß (IL-1ß), bactenecin 7.5 (BAC7.5), bactenecin 5 (BAC5), ß2-defensin (GBD2), hepcidin (HAMP), chemokine 4 (CCL4), tumour necrosis factor α (TNFα), toll-like receptor 2 (TLR2), cathelicidin-7 (MAP34) and cathelicidin-6 (MAP28) were determined in MSC. Milk, fat, and protein yields were higher and somatic cell count (SCC expressed as natural logarithm) was lower in the milk of goats fed organic Se. The Se concentration in milk was twice as high in the organic vs. inorganic treatment groups at the end of the experiment, while there were no differences in studied biochemical parameters between groups. The transcript levels of CSN1S2 and BAC7.5 were higher and IL-8 was lower in MSC of Se-yeast treated groups. Such results may indicate better health status of mammary glands of goats treated with organic Se as well as positive impact of selenized yeast on the goat's milk composition. Differences in the IL-1ß and IL-8 transcript levels were also noted between the stages of lactation, with the highest expression at the peak of lactation (day 70), highlighting the metabolic burden at this time. We concluded that the Se-yeast supplementation improved the productivity and health status of goats and could have significant economic impact on farmer's income.

MLST and RAPD molecular analysis of Staphylococcus aureus subsp. anaerobius isolated from goats in Poland.

Staphylococcus aureus subsp. anaerobius is an etiological agent of Morel's disease in small ruminants. The infection results in superficial abscesses located near lymph nodes. In the study, molecular analysis based on multilocus sequence typing (MLST) of seven housekeeping genes (arcC, aroE, glp, gmk, pta, tpi, yqiL) and random amplified polymorphic DNA (RAPD) was carried out on 19 S. aureus subsp. anaerobius strains isolated from two different goat herds from Poland. All of the 19 S. aureus subsp. anaerobius strains were found to belong to single MLST and RAPD types which support the high clonality level of this agent. However, the results obtained show clearly that the S. aureus subsp. anaerobius clone found in goats in Poland is different from those previously described. However, it is identical to the ATCC 38844 strain isolated from sheep in Spain, which has not been so far genotyped using MLST.

Small ruminant lentivirus infection influences expression of acute phase proteins and cathelicidin genes in milk somatic cells and peripheral blood leukocytes of dairy goats.

The aim of the study was to analyze acute phase protein and cathelicidin gene responses to small ruminant lentivirus (SRLV) infection in goats. In uninfected goats, we found higher Cp and lower Fbγ mRNA levels in blood leucocytes (BL) than in milk somatic cells (MSC), as well as lower SAA, Hp, and CRP and higher Cp and AGP concentrations in blood serum than in milk. In SRLV-infected goats, we found higher Fbγ and MAP28 and lower Cp expression in MSC than in BL, and higher SAA, Hp, Fb, and MAP28 and lower AGP concentrations in milk than in blood serum. Higher SAA and Hp expressions in BL and Hp expression in MSC were found in SRLV-infected goats. In SRLV-infected goats, we observed a higher concentration of SAA in blood serum, while in milk, lower SAA, Cp, and MAP28 and higher MAP34 concentrations were observed. The expression profiles of the studied genes differed between BL/serum and MSC/milk. The elevated SAA concentration in blood serum was accompanied by a decreased concentration of SAA and Cp in the milk of infected goats. No differences in the expression of the other studied genes may mean that the SRLV has the ability to evade the immune system, continuing to replicate. The elevated concentration of SAA in blood serum may promote viral multiplication. This higher concentration of SAA in blood serum and simultaneous reduced concentration of SAA and Cp in milk may be additive indicators of this infection.

Herd-level seroprevalence of Fasciola hepatica and Ostertagia ostertagi infection in dairy cattle population in the central and northeastern Poland.

BACKGROUND: Fasciola hepatica and Ostertagia ostertagi infections are widespread in cattle population of Europe, however data on their prevalence in Poland are only fragmentary. Therefore, the cross-sectional study was carried out to determine the herd-level seroprevalence of F. hepatica and O. ostertagi infection in dairy cattle population in the central and north-eastern provinces Poland, and to identify basic local risk factors for these infections. In total, 598 herds were enrolled, 394 (65.9%) in the north-eastern province and 204 (34.1%) in the central province. In each herd the questionnaire survey was conducted and bulk-tank milk (BTM) sample was collected and screened using two indirect immunoenzymatic tests. Optical density ratio (ODR) was regarded as the quantitative proxy of exposure to either of the two parasites. RESULTS: Both Fasciola and Ostertagia ELISA ODR in the north-eastern province was significantly higher than ODR in the central province. At the cut-off value of ODR = 0.27 the true herd-level seroprevalence of F. hepatica was 79.6% (95% CI: 74.0%, 84.3%) in the north-eastern province and 13.0% (95% CI: 5.3%, 21.7%) in the central province. At the cut-off of ODR = 0.50151 of 188 herds (80.3%, 95% CI: 74.1%, 85.4%) were seropositive for O. ostertagi in the north-eastern province and only 70 of 136 herds (51.5%, 95% CI: 43.1%, 59.7%) were seropositive in the central province. Location of a herd in the north-eastern province, longer grazing period practiced in a herd and > 50%-share of grazing grass in roughage were all positively related to the increase in exposure to both parasites. Moreover, the use of hay or haylage as main roughage proved to be positively related to the increase in exposure to F. hepatica. CONCLUSIONS: F. hepatica and O. ostertagi are widespread in cattle population in Poland, however their occurrence at a herd-level varies between different regions of Poland. This diversity can only partly be explained by different herd management, and appears linked to environmental and climate conditions typical for these regions.

Influence of small ruminant lentivirus infection on cheese yield in goats.

Three-year cohort study was carried out to investigate the influence of small ruminant lentivirus (SRLV) infection on cheese yield in goats. For this purpose records of milk yield, milk composition and cheese yield were collected in a dairy goat herd. Cheese yield was recorded as the amount of fresh cheese obtained from 1 kg milk. All goats were serologically tested for SRLV infection twice a year. The analysis included 247 records in total (71 for seropositive and 176 from seronegative individuals) and was carried out with the use of the four-level hierarchical linear model (α = 0·05). SRLV infection proved to be a statistically significant independent factor reducing cheese yield (P = 0·013)--when other covariates were held constant cheese yield was reduced by 4·6 g per each 1 kg milk in an infected goat compared with an uninfected goat. Other statistically significant covariates positively associated with cheese yield were protein contents, fat contents and the 3rd stage of lactation (P < 0·001 for all).

Seropositive bucks and within-herd prevalence of small ruminant lentivirus infection.

Caprine arthritis-encephalitis is an economically important disease of goats. It is evident that horizontal transmission through respiratory secretions and milk plays an important part in the disease spread whereas the role of sexual transmission remains questionable. The cross-sectional study was carried out to investigate the relationship between presence of small ruminant lentivirus (SRL V)-seropositive bucks and seroprevalence of SRL V infection in does in herds. The analysis included 76 goat herds seropositive for SRL V infection. A sample of adult female goats from each herd was selected in a simple random fashion. All males present in a herd were also enrolled in the study. The animals were screened with commercial serological immunoenzymatic tests. Standardized questionnaires were used to gather knowledge of 3 hypothesized herd-level confounding factors: number of years for which a herd had existed until testing, goat replacement from other herds in Poland and use of machine milking. Three-level hierarchical linear regression model was developed to evaluate the relationship (α = 0.05). Median (interquartile range) within-herd seroprevalence of SRL V was 60.1% (35.7% to 87.9%) and 35.8% (10.1% to 49.6%) in herds where seropositive males were present and absent, respectively. Controlling for possible confounders presence of SRL V-seropositive bucks proved to be an independent factor linked to the higher within-herd seroprevalence of SRL V (p = 0.001). The study indicates that seropositive bucks may facilitate the spread of SRL V infection in goat herds and therefore their presence should be considered as a risk factor.

Morphometry of the heart orifices and morphometry and topography of the coronary ostia in the goat.

BACKGROUND: The coronary vessels have been described in various species of domestic and wild ruminants. However, no studies on the detailed morphology and morphometry of heart orifices and coronary ostia in the Polish goat are available. MATERIALS AND METHODS: The study was carried out on 112 female, adult dairy goats belonging to Polish Fawn Improved and Polish White Improved breed, closely related to French Alpine and Saanen, respectively. RESULTS: In all examined individuals, all heart orifices and heart valves were of normal structure. There was no significant difference between diameter of the aortic and pulmonary orifice. The right atrioventricular opening was significantly wider than the left atrioventricular opening. The dimension of the left coronary ostium ranged from 1.0 to 5.5 mm with the arithmetic mean (± standard deviation [SD]) of 4.3 ± 0.8 mm. The dimension of the right coronary ostium ranged from 0.5 to 5.0 mm with the arithmetic mean (± SD) of 2.8 ± 0.7 mm. Both coronary artery ostia were located under the sinotubular junction. Out of 112 examined goats, 39 (34.8%) had variations in the structure of the coronary ostia such as the lack of main trunk or the presence of additional coronary ostia. They were observed in one (in 34/39 goats) or both coronary arteries (5/39 goats). CONCLUSIONS: In goats, the dimensions of aortic and pulmonary orifices are similar while the right atrioventricular opening outsizes the left one. On the other hand, the left coronary ostium is wider than the right one. Morphological variations in the coronary ostia occur in approximately one third of goats, more often in the right than in the left ostium.

Herd-level true seroprevalence of caseous lymphadenitis and paratuberculosis in the goat population of Poland.

A large-scale study was carried out in the Polish goat population in 2014-2021 to determine the herd-level true seroprevalence (HTP) of caseous lymphadenitis (CLA) caused by Corynebacterium pseudotuberculosis (Cp) and paratuberculosis (PTB) caused by Mycobacterium avium ssp. paratuberculosis (Map). Two-stage cluster sampling was applied to herds counting at least 20 adult goats (aged >1 year) and in each herd all males and 10-13 females were tested. At least one seropositive goat regardless of its sex was necessary to consider the herd as infected. HTP was estimated using the Bayesian approach with the Gibbs sampler in the EpiTools and reported as the median and 95 % credibility interval (95 % CrI). A total of 1282 adult goats from 86 herds were serologically tested using two commercial ELISAs (Cp-ELISA and Map-ELISA). At least 1 seropositive result of Cp-ELISA and Map-ELISA was obtained in 73/86 herds (84.9 %) and 40/86 herds (46.5 %), respectively. HTP of CLA was estimated at 73.3 % (95 % CrI: 65.0 %, 80.4 %) and HTP of PTB was estimated at 42.9 % (95 % CrI: 25.8 %, 58.0 %). There was a significant positive association between the occurrence of CLA and PTB in the herds (odds ratio 6.0, 95 % confidence interval: 1.2, 28.8; p = 0.010). Probability of the seropositive result for PTB was also significantly higher in Cp-seropositive goats than in Cp-seronegative goats (odds ratio 3.9, 95 % confidence interval: 2.4, 6.3; p < 0.001) which could indicate either a higher risk of co-infection or a higher rate of false positive results for PTB in Cp-positive goats. To investigate this issue, optical densities obtained in Map-ELISA were compared between Cp-positive and Cp-negative goats and results of Map-ELISA were adjusted accordingly. Map-negative sera from Cp-positive goats turned out to have significantly higher optical densities than Map-negative sera from Cp-negative goats (p < 0.001). After the adjustment, the herd-level apparent seroprevalence of PTB was 41.9 % (36/86 herds) so it still fell within the 95 % CrI of HTP of PTB calculated before the adjustment. Concluding, CLA appears to be widespread in the Polish goat population. In many of them it may be subclinical at the moment, however will likely emerge in the future as the disease follows cyclic pattern in Poland. On the other hand, given the total lack of clinical PTB in Polish goats, an explanation for a high HTP of PTB remains unclear and warrants further studies using tests of higher analytical specificity than ELISA.

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.

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.

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.

Correlation between metabolomic profile constituents and feline pancreatic lipase immunoreactivity.

BACKGROUND: Feline pancreatic lipase immunoreactivity (fPLI) is commonly used to diagnose pancreatitis in cats (FP). Untargeted metabolomics has been extensively applied in human and veterinary medicine, but no metabolomic studies regarding FP have been conducted. OBJECTIVES: To identify metabolites significantly associated with increased fPLI. ANIMALS: Forty-nine client-owned cats: 11 clinically healthy and 38 with various clinical conditions. METHODS: Analytical cross-sectional study with convenience sampling. A panel of 630 metabolites belonging to 26 biochemical classes was quantified in plasma using a commercial metabolomic assay. The correlation between plasma metabolite concentrations and serum fPLI was evaluated using Spearman's rank correlation coefficient (Rs ) with Bonferroni correction. Multivariable analysis then was performed to control for glomerular filtration rate, liver damage, and blood glucose concentration. The accuracy of selected metabolites in discriminating between cats with normal (≤3.5 µg/L) and increased (>5.3 µg/L) fPLI was estimated using the area under the receiver operating characteristic curve (AUROC). RESULTS: Four hundred and seven of 630 metabolites (64.6%) were quantified in all cats. When controlled for potential confounders only 3 sphingolipids were significantly positively correlated with fPLI: 2 cerebrosides: HexCer(d18:1/24:0); (Rs  = .56), and HexCer(d18:1/24:1); (Rs  = .58) and 1 sphingomyelin: SM C18:0 (Rs  = .55). Their AUROCs in identifying cats with increased fPLI were 82% (95% confidence interval [CI 95%], 70%-94%), 84% (CI 95%, 72%-96%), and 78% (CI 95%, 65%-92%), respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: Selected sphingolipids are moderately positively correlated with fPLI and appear to have fair to moderate diagnostic accuracy in discriminating between cats with normal and increased fPLI.

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.

Accuracy of acute-phase proteins in identifying lethargic and anorectic cats with increased serum feline pancreatic lipase immunoreactivity.

The mainstay of laboratory diagnostics of feline pancreatitis (FP) is measuring serum feline pancreatic lipase immunoreactivity (fPLI). Thus far, several studies have investigated the relationship between acute-phase proteins (APPs) and the presence and severity of FP. Although changes in serum amyloid A (SAA) concentration have been linked with FP, its diagnostic accuracy remains unknown. We aimed to determine the accuracy of selected APPs in identifying cats with a high risk of FP based on increased fPLI. Serum fPLI was measured in cats (n = 52) that were presented to veterinary clinics with signs of lethargy and anorexia. The cats were divided into two groups (fPLI ≤3.5 µg/L; n = 27 and fPLI >3.5 µg/L, n = 25), corresponding to low and high risks of FP. Serum albumin, globulin, haptoglobin (Hp), and SAA concentrations, as well as the albumin-to-globulin ratio (A/G) and SAA-to-albumin (SAA/A) ratios, were determined and compared between groups. The accuracy of these measurements was examined using a multivariable logistic regression model and receiver operator characteristic (ROC) curve analysis. SAA concentrations and SAA/A ratios were significantly (P = .026 and P = .022) higher in cats with increased fPLI, with the area under ROC curve (AUROC) of 68.1% (CI 95%: 53.4%, 82.9%) and 68.6% (CI 95%: 54.0%, 83.2%), respectively. The logistic regression model combining SAA and A/G had AUROC of 75.2% (CI 95%: 62.0%, 88.4%) for identifying cats with increased fPLI. SAA as a sole analyte or combined with A/G had low to moderate accuracy in identifying anorexic, lethargic cats with increased fPLI. Serum albumin, globulin, and Hp concentrations had no discriminatory potential in these cats.