Mastitis in Dairy Cattle: On-Farm Diagnostics and Future Perspectives.

Mastitis is one of the most important diseases in dairy cattle farms, and it can affect the health status of the udder and the quantity and quality of milk yielded. The correct management of mastitis is based both on preventive and treatment action. With the increasing concern for antimicrobial resistance, it is strongly recommended to treat only the mammary quarters presenting intramammary infection. For this reason, a timely and accurate diagnosis is fundamental. The possibility to detect and characterize mastitis directly on farm would be very useful to choose the correct management protocol. Some on-field diagnostic tools are already routinely applied to detect mastitis, such as the California Mastitis Test and on-farm culture. Other instruments are emerging to perform a timely diagnosis and to characterize mastitis, such as Infra-Red Thermography, mammary ultrasound evaluation and blood gas analysis, even if their application still needs to be improved. The main purpose of this article is to present an overview of the methods currently used to control, detect, and characterize mastitis in dairy cows, in order to perform a timely diagnosis and to choose the most appropriate management protocol, with a specific focus on on-farm diagnostic tools.

Metabolic Changes Associated with Different Levels of Energy Deficits in Mediterranean Buffaloes during the Early Lactation Stage: Type and Role of the Main Lipid Fractions Involved.

Cell function and energy redistribution are influenced by lipid classes (phospholipids (PLs), free fatty acids (FFAs), triglycerides (TGs), and cholesterol esters (CEs)). The aim of this study was to investigate metabolic alterations that are related to changes in lipid classes according to different levels of energy deficits in early lactating Mediterranean buffaloes (MBs). Sixty-three MBs were enrolled at the beginning of lactation using an observational study with a cross-sectional experimental design. Serum ß-hydroxybutyrate (BHB) levels were used to group the animals into a healthy group (Group H; n = 38; BHB < 0.70 mmol/L) and hyperketonemia risk group (Group K; n = 25; BHB ≥ 0.70 mmol/L). Statistical analysis was performed using a linear model that included the effect of the group and body condition score to assess differences in fatty acid (FA) concentrations. A total of 40 plasma FAs were assessed in each lipid class. Among the FAs, eight PLs, seven FFAs, four TGs, and four CEs increased according to BHB levels, while three FFAs, three TGs, and one CE decreased. The changes among lipid class profiles suggested the influence of inflammatory response, liver metabolism, and the state of body lipid reserves. In addition, the possible similarities of buffaloes at risk of hyperketonemia with ketotic cows suggest the necessity of further investigations in these ruminants.

Salmonella Enteritidis Fatal Septicemia with Meningoencephalitis in a Tiger (Panthera tigris) Cub.

A 15-day-old, female, captive Panthera tigris cub was hospitalized after developing severe hyperthermia, depression, and lack of appetite. The clinical condition rapidly worsened, and the tiger cub died in 72 h after the onset of neurological symptoms, septic shock, and multiple organ dysfunction syndrome. The postmortem main gross findings consisted of a severe and diffuse bilateral fibrino-suppurative meningoencephalitis and ventriculitis, mild fibrinous and sero-hemorrhagic polyserositis and cystitis, severe pulmonary edema, and hemorrhages. Microscopically, the meninges, ependyma, and choroid plexuses were diffusely expanded by abundant infiltration of neutrophils and macrophages, with multifocal fibrinous exudation. Histiocytic interstitial pneumonia, fibrinous and neutrophilic polyserositis, and pyelocystitis were also observed. Vascular thrombosis with multifocal vasculitis and vascular necrosis were frequently observed. Aerobic and anaerobic cultures performed on the brain, lungs, intestine, kidneys, and in pericardial effusion reported the presence of Salmonella enterica subsp. enterica serovar Enteritidis. Environmental and nutritional contamination were identified as putative sources of infections. To the best of the authors' knowledge, this is the first report of Salmonella Enteritidis septicemia with meningoencephalitis in a tiger cub, which highlights the need to further investigate the cause of acute perinatal death to reduce the risk of infectious disease outbreaks.

Evaluation of the metabolomic profile through 1H-NMR spectroscopy in ewes affected by postpartum hyperketonemia.

Ketosis is one of the most important health problems in dairy sheep. The aim of this study was to evaluate the metabolic alterations in hyperketonemic (HYK) ewes. Forty-six adult Sardinian ewes were enrolled between 7 ± 3 days post-partum. Blood samples were collected from the jugular vein using Venosafe tubes containing clot activator from jugular vein after clinical examination. The concentration of ß-hydroxybutyrate (BHB) was determined in serum and used to divide ewes into assign ewes into: Non-HYK (serum BHB < 0.80 mmol/L) and HYK (serum BHB ≥ 0.80 mmol/L) groups. Animal data and biochemical parameters of groups were examined with one-way ANOVA, and metabolite differences were tested using a t-test. A robust principal component analysis model and a heatmap were used to highlight common trends among metabolites. Over-representation analysis was performed to investigate metabolic pathways potentially altered in connection with BHB alterations. The metabolomic analysis identified 54 metabolites with 14 different between groups. These metabolites indicate altered ruminal microbial populations and fermentations; an interruption of the tricarboxylic acid cycle; initial lack of glucogenic substrates; mobilization of body reserves; the potential alteration of electron transport chain; influence on urea synthesis; alteration of nervous system, inflammatory response, and immune cell function.

Changes in plasma fatty acids profile in hyperketonemic ewes during early lactation: a preliminary study.

The transition from late pregnancy to early lactation is characterized by marked changes in energy balance of dairy ruminants. The mobilization of adipose tissue led to an increase in plasma non-esterified fatty acids (NEFA) and ß-hydroxybutyrate (BHB). The aim of this study was to analyze the total plasma fatty acids of healthy and hyperketonemic dairy ewes in early lactation through gas chromatography (GC) to evaluate metabolic alterations. An observational study was used with a cross-sectional experimental design. Forty-six Sarda dairy ewes were enrolled in the immediate post-partum (7 ± 3 days in milk) and divided into two groups according to serum BHB concentration: non-hyperketonemic group (n = 28; BHB < 0.86 mmol/L) and hyperketonemic group (n = 18; BHB ≥ 0.86 mmol/L). A two-way ANOVA included the effect of group and parity was used to evaluate differences in fatty acids (FA) concentrations. A total of 34 plasma FA was assessed using GC. 12 out of 34 FA showed a significant different between groups and 3 out of 34 were tended to significance. Only NEFA concentration and stearic acid were influenced by parity. The results may suggest possible links with lipid metabolism, inflammatory and immune responses in hyperketonemic group. In conclusion, GC represents a useful tool in the study of hyperketonemia and primiparous dairy ewes might show a greater risk to develop this condition.

Development of an equation to screen for solar hemorrhages from digital cushion ultrasound texture analysis in veal calves at slaughter.

Claw disorders are a relevant welfare issue in the cattle industry, fast and accurate diagnoses are essential for successful treatment and prevention. The present study aimed to develop an equation to assess the presence of solar hemorrhages from real-time ultrasound images texture analysis at slaughter. Eighty-eight hind feet were collected at the slaughterhouse from 44 Holstein male veal calves. The claws were trimmed by a veterinarian hoof-trimmer, approximately 30 min after the calves' slaughter, and classified into healthy and affected by solar hemorrhages. At the same time, ultrasound images were collected for each claw. Sole soft tissues' thickness was measured, and texture analysis was performed using MaZda software. The resulting parameters from sole soft tissues' measurements and texture analysis were screened with a stepwise linear discriminant analysis using the absence or presence (0/1) of solar hemorrhages as the dependent variable. Results from the stepwise analysis identified 9 variables (among 279) as predictors, and an equation was developed and used to predict the presence or absence of solar hemorrhages on the scanned claws by binary measure: values ≤0.5 counted as 0, while those >0.5 as 1. Validation of the equation was performed by testing predicted lesions (LESpred) against the clinically evaluated lesions (LESeval) with a confusion matrix, a ROC analysis, and a precision-recall curve. Results of the present study suggest that the equation proposed has a good potential for detecting effectively hemorrhages of the sole by ultrasound imaging texture means, and could be used to monitor unsatisfactory housing and management conditions at the farm level, and for early management intervention and prevention.

Evaluation of body surface temperature variations in dogs affected by spinal cord injuries during physiotherapy exercise in a water treadmill.

The aim this study was to evaluate variation in body surface temperature (BST) in healthy and spinal cord injured (SCI) dogs, and to outline temperature variation at rest (T0), during (T1) and after (T2) water-treadmill physiotherapy sessions in SCI using infrared thermography (IRT). Sixty-seven dogs of different sex, breed, body weight and age were enrolled: 14 healthy dogs and 53 dogs affected by disc pathologies. The study examined three regions of interest (ROIs): the total image of the spine (IMAGE), the spinal cord area from 1st thoracic vertebra to the last lumbar vertebra (AR01) and the surgery wound or spinal cord lesion area (AR02). Significant BST variations between healthy and SCI were reported in T°max and T°max-min (ΔT) values in IMAGE (P < 0.05). In SCI group, AR01 and AR02 assessment showed an increase in temperature ate the sited of the injured area and adjacent body structures. In SCI, a significant effect of water-treadmill exercise in AR01 and AR02 was reported. In fact, both AR01 and AR02 reported higher BST (T°max, T°mean, T°min and ΔT) during the physical exercise (T1), representing the response to physical activity of the spine vascularization, muscles and column contiguous tissues. Furthermore, in T2, the same areas reported a significant lower BST (T°max, T°mean, and ΔT), related to a decrease in tissue inflammation on the target area of the water treadmill physiotherapy. This study highlights how IRT can detect BST variations associated with injured areas. In addition, IRT revealed a positive effect of water-treadmill exercise on the injured spinal cord areas, thus it could be a viable non-invasive and rapid method to support both clinical examination and assessment of the effectiveness of medical treatment in SCI.

Lung Ultrasonography and Clinical Follow-Up Evaluations in Fattening Bulls Affected by Bovine Respiratory Disease (BRD) during the Restocking Period and after Tulathromycin and Ketoprofen Treatment.

Bovine respiratory disease (BRD) is a global infectious disease, causing decreased well-being and economic losses in livestock, frequently during the restocking period. The aim of this study was to evaluate the feasibility of thoracic ultrasonography (TUS) to assess BRD in restocked animals, and the effectiveness of tulathromycin and ketoprofen treatment in sick animals. A total of 60 Limousin fattening bulls were enrolled. On the day of restocking (T0), the animals were divided into two groups based on TUS assessment of six lung areas: group C (ultrasonography score (US score) < 3) and group D (US score ≥ 3). Group D received a single administration of tulathromycin and ketoprofen at T0: this group was revaluated after 1.5, 3, 7, and 14 days. Both groups were revaluated after 21 days. The two groups showed a significant difference both in US score and clinical symptoms (respiratory score, nasal and ocular discharges, and rectal temperature) at T0. In group D, the treatment was effective in improving the clinical symptoms and US score, particularly reducing the severity of lung lesions. TUS represents a non-invasive and cost-effective tool for BRD early diagnosis and for monitoring treatment efficacy in restocked livestock.

Differences in the serum metabolome profile of dairy cows according to the BHB concentration revealed by proton nuclear magnetic resonance spectroscopy (1H-NMR).

The mobilization of body reserves during the transition from pregnancy to lactation might predispose dairy cows to develop metabolic disorders such as subclinical ketosis or hyperketonemia. These conditions are not easily identifiable and are frequently related to other diseases that cause economic loss. The aim of this study was to evaluate the serum metabolome differences according to the ß-hydroxybutyrate (BHB) concentration. Forty-nine Holstein Friesian dairy cows were enrolled between 15 and 30 days in milk. According to their serum BHB concentration, the animals were divided into three groups: Group 0 (G0; 12 healthy animals; BHB ≤ 0.50 mmol/L); Group 1 (G1; 19 healthy animals; 0.51 ≤ BHB < 1.0 mmol/L); and Group 2 (G2; 18 hyperketonemic animals; BHB ≥ 1.0 mmol/L). Animal data and biochemical parameters were examined with one-way ANOVA, and metabolite significant differences were examined by t-tests. Fifty-seven metabolites were identified in the serum samples. Thirteen metabolites showed significant effects and seemed to be related to the mobilization of body reserves, lipids, amino acid and carbohydrate metabolism, and ruminal fermentation.

Differences of the Plasma Total Lipid Fraction from Pre-Foaling to Post-Foaling Period in Donkeys.

The period from late gestation to early lactation is characterized by changes in energy metabolism. The aim of the current study was to analyze the plasma total lipid fraction using gas chromatography (GC) analysis, in order to highlight metabolic changes from the pre-foaling to post-foaling periods. Eleven pluriparous dairy jennies (mean age of 11.88 ± 3.79 years) belonging to the Amiata donkey breed were enrolled. Blood sampling was performed at 15 days before foaling (T0), and 15 (T1), 30 (T2), 60 (T3), and 90 (T4) days after foaling, for biochemical and GC analysis. A total of 37 fatty acids were identified in plasma samples: 4 medium chain (MCFA), 24 long chain (LCFA), and 9 very-long chain (VLCFA) fatty acids. Among them, 20 fatty acids changed significantly, and two fatty acid showed a trend toward significance. Furthermore, the LCFA, saturated, unsaturated, monounsaturated, and polyunsaturated ω-3 fatty acids changed significantly during the study period. The main alterations were between T0 and the other time points and appeared to be related to lipid metabolism, cellular structure and function, and inflammatory and immune responses. Our findings reveal greater energy requirements at the end of gestation compared to early lactation in donkeys.

Thermographic Screening of Beef Cattle Metatarsal Growth Plate Lesions.

Lameness represents one of the main causes of decreased productive performance and impaired animal welfare in the bovine industry. Young beef bulls are predisposed to develop diseases of the growing skeleton, especially growth plate lesions. Early diagnosis is indispensable for ensuring correct treatment, fast recovery and reduction losses. However, when dealing with beef cattle, this is not always possible. Fast and reliable diagnostic imaging techniques are necessary to improve dealing with lameness in beef animals. The aim of the present study was to examine the potential of thermographic imaging as a non-invasive tool for rapidly screening beef bulls for the presence of growth plate lesions. Here, 20 Charolais and Limousine beef bulls affected by growth plate lesions in one of the rear limbs were selected. IRT was performed on both hind limbs using a digital infrared camera (ThermaCam T420 Model, Flir Systems, Boston, MA, USA), prior to radiographic imaging and clinical examination. The temperature of healthy and affected limbs was measured in two regions: the area correspondent to the growth plate (AR01) and the whole area of the metatarsus (AR02). Growth plate lesions were found to increase the maximum, mean, and minimum temperatures in AR01; and the mean and maximum temperatures in AR02, therefore, indicating the potential of IRT as a reliable, practical tool for screening growth plate lesions in beef bulls.

Associations between ultrasound measurements and hematochemical parameters for the assessment of liver metabolic status in Holstein-Friesian cows.

Metabolic disorders, including hepatic lipidosis and ketosis, severely affect animal health status and welfare with a large economic burden in dairy herds. The gold standard for diagnosing hepatic lipidosis is the liver biopsy, which is impractical and invasive for the screening at farm level. Ultrasound (US) imaging is a promising technique for identifying liver dysfunction, but standardized specifications in physiological conditions are needed. Herein, we described the features of four US measurements, namely the liver predicted triacylglycerol (pTAG) content, liver depth (LD), and portal vein area (PVA) and depth (PVD) and we investigated their associations with a set of hematochemical (HC) indicators in 342 clinically healthy Holstein Friesian dairy cows. Liver pTAG content was negatively associated with hematocrit and positively with globulin, whereas PVA was negatively associated with thiol group levels, and LD positively with ceruloplasmin. We found significant interactions between some HC parameters and parity: in particular, creatinine, thiol groups and globulin for PVA, and aspartate aminotransferase, paraoxonase and ceruloplasmin for PVD. This study offers new insights on variations in liver function occurring after calving and pave the way for the potential use of minimally invasive techniques for prompt detection of metabolic disorders in dairy herds.

Milk Fatty Acids Composition Changes According to ß-Hydroxybutyrate Concentrations in Ewes during Early Lactation.

Ketosis is a metabolic disease of pregnant and lactating ewes linked to a negative energy balance which can cause different economic losses. The aims of this study were to understand the metabolic status of the early-lactating ewes and to identify biomarkers for early diagnosis of subclinical ketosis. Forty-six Sarda ewes were selected in the immediate post-partum for the collection of the biological samples. A blood sample from the jugular vein was used to determine ß-Hydroxybutyrate (BHB) concentrations. Animals were divided into two groups: BHB 0 or healthy group (n = 28) with BHB concentration < 0.86 mmol/L; and BHB 1 or subclinical ketosis (n = 18) with a BHB concentration ≥ 0.86 mmol/L. Ten mL of pool milk were collected at the morning milking for the analyses. The concentration of 34 milk fatty acids was evaluated using gas chromatography. Two biochemical parameters and 11 milk fatty acids of the total lipid fraction presented a p-value ≤ 0.05. The study revealed different relationships with tricarboxylic acid cycle, blood flows, immune and nervous systems, cell functions, inflammatory response, and oxidative stress status. Eight parameters were significant for the receiver operating characteristic (ROC) analysis with an area under the curve greater than 0.70.

Application of Ultrasound Images Texture Analysis for the Estimation of Intramuscular Fat Content in the Longissimus Thoracis Muscle of Beef Cattle after Slaughter: A Methodological Study.

Intramuscular fat (IMF) is a major trait in the evaluation of beef meat, but its determination is subjective and inconsistent and still relies on visual inspection. This research objective was a method to predict IMF% from beef meat using ultrasound (US) imaging texture analysis. US images were performed on the longissimus thoracis muscle of 27 Charolaise heifers. Cuts from the 12th to 13th ribs were scanned. The lipid content of the muscle samples was determined with the petrol ether (Randall) extraction method. A stepwise linear discriminant analysis was used to screen US texture parameters. IMF% measured by chemical extraction (IMFqa) was the dependent variable and the results of the texture analysis were the explanatory variables. The model highlighted seven parameters, as a predictive and a multiple regression equation was created. Prediction of IMF content (IMFpred) was then validated using IMFqa as ground truth. Determination coefficient between IMFqa and IMFpred was R2 = 0.76, while the ROC analysis showing a sensitivity of 88% and a specificity of 90%. Bland-Altman plot upper and lower limit were +1.34 and -1.42, respectively (±1.96 SD), with a mean of -0.04. The results from the present study therefore suggest that prediction of IMF content in muscle mass by US texture analysis is possible.

Comparison between two preventive treatments for hyperketonaemia carried out pre-partum: effects on non-esterified fatty acids, ß-hydroxybutyrate and some biochemical parameters during peripartum and early lactation.

The objective of this study was to compare the effect of two different preventive protocols, on serum ß-hydroxybutyrate (BHB) concentration and liver health indices pre-partum and during early-lactation in high-yielding Holstein dairy cows. One hundred cows were randomly divided into three groups: control group (CTRL, n = 20, without preventive treatment), second group (SUPP, n = 40 animals treated with a compound based on acetyl-methionine, inositol, cyanocobalamin, l-alanine, l-arginine, l-threonine, l-glutamic acid supplementation and α-lipoic acid) and third group (MON, n = 40 animals treated with monensin). Blood samples were collected from all cows at on 3 occasions pre-partum and 3 occasions post-partum. Body condition (BCS) score was evaluated and glucose, non-esterified fatty acids (NEFA), BHB, triglycerides, total cholesterol, alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyltransferase (GGT), total bilirubin, total proteins, globulins, albumin and urea concentrations were assessed. Two-way repeated measures analysis of variance was applied. Statistically significant differences among the three experimental groups were found in the values of all studied parameters (P < 0.05). Our results confirm the established beneficial effect of MON treatment in decreasing BHB levels and increasing glucose availability after calving. Serum biochemical analysis revealed the expected post-partum alterations attributable to adaptations that influenced the metabolism and liver function in CTRL, whereas these alterations were reduced or absent in SUPP and MON. Results from the present study suggest that both preventive protocols, but in particular SUPP, could positively affect selected indicators of energy metabolism reducing the risk of hyperketonaemia and increase of liver function in Holstein dairy cows, both pre- and post-partum.

A retrospective study on transabdominal ultrasound measurements of the rumen wall thickness to evaluate chronic rumen acidosis in beef cattle.

BACKGROUND: Chronic and subacute rumen acidosis are economically important in the beef industry. The aim of this study was to evaluate the potential suitability of the transabdominal ultrasonographic examination of the ruminal wall to diagnose chronic rumen acidosis in beef cattle compared to direct measurement of ruminal pH, as a fast non-invasive tool to be used in field condition. Ultrasonographic examination of the rumen was conducted in 478 beef cattle before rumenocentesis (chronic rumen acidosis group = pH ≤ 5.8; healthy group = pH ≥ 5.9). Rumen wall ultrasound measurements included rumen wall thickness (RWT) and rumen mucosa and submucosa thickness (RMST). RESULTS: The Analysis of Variance showed the high significant effect of the pH class for RWT and RMST (P < 0.001). Spearman RANK correlation analysis showed interaction between rumen pH and RWT (- 0.71; P < 0.0001) and RMST (- 0.75; P < 0.0001). A significant Spearman's correlations were found between volatile fatty acids (VFA) and RWT and RMST. The differentiation efficiency of RWT between healthy and chronic rumen acidosis groups, as a result of the receiver operator curve (ROC) analysis, was quite good with an area under the receiver operator curve (AUROC) of 0.88: P < 0.0001; 95% CI: 0.83-0.98. Using a cut-off value of > 8.2 mm. The differentiation efficiency of RMST between healthy and chronic rumen acidosis groups, as a result of ROC curve analysis, was good with an AUROC of 0.90: p < 0.0001; 95% CI: 0.85-0.94. Using a cut-off value of > 5.3 mm. CONCLUSIONS: In this study, the thickening of RWT and RMST is correlated with the changes of ruminal pH. Transabdominal rumen ultrasound has the potential to become a powerful diagnostic tool useful to identify fattening bulls affected by chronic rumen acidosis.

Changes of Plasma Fatty Acids in Four Lipid Classes to Understand Energy Metabolism at Different Levels of Non-Esterified Fatty Acid (NEFA) in Dairy Cows.

The transition period is a central moment in dairy cows breeding because metabolic disorders may occur in relation to a dramatic increase in energy demand. This research aimed to identify new biomarkers for the diagnosis of hyperketonemia in bovine in early lactation phase with different value of plasmatic non-esterified fatty acid (NEFA). The profile of plasma fatty acids (FAs) divided into four lipid classes was evaluated using thin layer chromatography and gas chromatographic techniques (TLC-GC). A group of 60 multiparous Holstein-Friesian dairy cows were recruited in the present study. Blood samples were collected from the coccygeal vein and NEFA and the ß-hydroxybutyrate (BHB) were evaluated. All animals were divided in 2 groups based on NEFA, NEFA0 group had as mean value 0.24 ± 0.12 mEq/L and NEFA1 group had as mean value 0.87 ± 0.23 mEq/L. Plasma FA concentrations were analyzed separately in free fatty acids, cholesterol esters, phospholipids and triglycerides. Six FAs demonstrated a predictive value in the hyperketonemic dairy cows. In the free fatty acid class, the predictive FAs were C14:0 (AUC = 0.77), C18:1 ω 9 (AUC = 0.72), C18:1 ω 7 (AUC = 0.70) and C18:3 ω 3 (AUC = 0.68). In the phospholipids class the predictive parameters were C12:0 (AUC = 0.78) and C8:0 (AUC = 0.73). In cholesterol, esters and triglycerides lipidic classes no FA had a predictive function.

Changes of milk fatty acid composition in four lipid classes as biomarkers for the diagnosis of bovine ketosis using bioanalytical Thin Layer Chromatography and Gas Chromatographic techniques (TLC-GC).

The aim of this study was to extend the limited research available on the association between the concentration of milk fatty acids and the elevated plasmatic value of ß-hydroxybutyrate (BHB) in early lactation of dairy cows. Fifty-four Holstein Friesian dairy cows were enrolled in the study. All animals were classified on the basis of their blood BHB concentration: BHB ≥ 1.0 mmol/L (BHB-1, sick group) and BHB ≤ 0.99 mmol/L (BHB-0, healthy group). Using Thin Layer Chromatography (TLC), four lipid classes (cholesterol esters -CE-, phospholipids -PL-, free fatty acids -FFA- and triacylglycerols -TAG-) were separated, and then the fatty acid (FA) composition was determined by High Resolution Gas Chromatography coupled with Flame Ionization Detector/Mass Spectrometer (HRGC-FID/MS). The FA profiles were used to investigate the diagnostic potential value of milk fatty acids for the correct classification of cows with BHB concentration above the established threshold (BHB < 1.0 mmol/L). Boruta Test and Receiver Operating Characteristic curves (ROC) were used to identify which FA and their thresholds of concentration could be used when animals presented hyperketonemia. The research has identified fourteen FA, belonging to CE, FFA, and TAG classes, useful for an association with BHB-1. These compounds, with predictive value for the development of hyperketonemia, could be considered valuable biomarkers. Further studies on a wider sampling, based on clinical and therapeutic approach, will be necessary to confirm, by bioanalytical chromatographic approaches, if these predictive FA will change between healthy and sick animals. New approaches in relation on the administration of different diets or supplements, and administration of drugs might improve the prevention of hyperketonemia.

Application of texture analysis of b-mode ultrasound images for the quantification and prediction of intramuscular fat in living beef cattle: A methodological study.

Intramuscular fat (IMF) contributes significantly to the aroma and tenderness of the meat, therefore playing a key role in quality determination. Yet, IMF determination methods rely on visual inspection or on fat extraction from meat samples after animals' slaughter. The aim of this methodological study was the elaboration of a process capable of predicting IMF% using real-time ultrasound (RTU) images in live beef cattle. The longissimus dorsi (LD) muscle of 26 Charolaise heifers was investigated. In vivo ultrasound images were taken and texture analysis was performed. One week after the animals' slaughter, the whole twelfth rib cut was collected, and IMF% was determined by extraction with petrol ether (Randall) method. Animals were divided in 3 groups depending on their mean lipid content percentage in 100 g meat (Group 1: IMF ≤ 4.24%; Group 2: 4.25% ≤ IMF ≤ 5.75%; Group 3: IMF ≥ 5.76%). Texture parameters were selected by a stepwise linear discriminant analysis using IMF% measured by chemical extraction (IMFqa) as the dependent variable, and the results of the texture analysis as explanatory variables. 6 variables were found predictive and molded into a multiple regression equation, this equation was then validated using IMFqa as ground truth. A high linear correlation between IMFqa and IMFpred was evident (r2 = 0.8504), ROC analysis perfomed on IMFpred comparing it to IMFqa showed a sensitivity of 80% and a specificity of 93.7%, while results from the Bland-Altman plot were ± 1.96 (±1.11SD).

Ultrasonographic alterations of bovine claws sole soft tissues associated with claw horn disruption lesions, body condition score and locomotion score in Holstein dairy cows.

Claw Horn Disruption Lesions (CHDL) negatively affect the sole soft tissue structures located beneath the sole horn. The aim of the present study was to investigate the effect of CHDL on sole soft tissues by ultrasound means, correlating Body condition score (BCS), locomotion score and CHDL with ultrasonography evaluations of sole soft tissues in Holstein dairy cows. 100 Holstein dairy cows were enrolled in the study. BCS and locomotion score were assessed and functional trimming was performed on all animals. 84 healthy claws and 174 claws with solely one CHDL per claw were evaluated both clinically and with ultrasound, and CHDL were identified and recorded. Sole soft tissues thickness (mm) and echogenicity was determined, and ultrasonographic alterations, related to CHDL presence, where measured long their vertical (L1) and horizontal (L2) axis. Statistically significant (P < .001) differences were found in echogenicity between healthy claws and all the affected ones, with the healthy ones being mainly anechoic. Statistically significant (P < .001) differences were found for vertical (L1) and horizontal (L2) axis measures between the diverse CHDL, confirming ultrasonography as a useful tool to distinguish lesions and their extension by measuring L1 and L2. BCS had an influence both on sole soft tissues ultrasonographic appearance and on CHDL insurgency. These results confirm ultrasonography as a reliable tool for detecting an increase in sole soft tissues echogenicity, that was seen to occur with CHDL insurgency, and in determining lesion extension.