Assessing teat canal morphology in the dry period and during lactation by high-resolution ultrasound.

Our objectives were to quantify the dimensions of a fully 'closed' teat canal in dry cows and to describe recovery of the teat canal between milkings in lactating cows to assess whether and when full closure is attained, since this is an important determinant of udder health. Using an ultrasound scanner, teat canal length and diameter (proximal, midpoint and distal), teat cistern width, teat end width, whole teat width and teat wall thickness in 77 dry and 39 lactating dairy cows were measured. The dry cows represented a cross section of the dry population, with days since dry off ranging from 0 to 69 (median: 27). Data from lactating cows were recorded just before milking, and every 3 h post-milking. To control for location a cross-over (parlour vs. barn) study design was used. In dry cows, teat canal length and diameter did not vary by quarter or days since dry off, but multiparous cows had significantly wider teat canals than primiparous cows. The dry cow measurements can be used as baseline for dimensions for closed teats. In lactating cows, all teat dimensions except teat end width changed significantly during the 12-h milking cycle. Location (parlour vs. barn) did not affect the measurements, except teat end width and teat wall thickness. Teat canal length increased after milking and returned to pre-milking values by 9 h. Proximal and midpoint teat canal diameters decreased slightly just after milking and then progressively increased to above the pre-milking values by 9 h. Distal teat canal diameter increased after milking, partially contracting by 9 h. We found that during the dry period the teat canal is in a steady state, but its diameter is not zero, while during the lactation, the teat canal is in a near constant state of remodelling.

Teat morphology across five buffalo breeds: a multi-country collaborative study.

The buffalo (Bubalus bubalis) is a species of worldwide importance, raised to produce milk, meat, and hides, and often used as a working animal in rural contexts with low access to hi-tech solutions. In the present study, 100 lactating buffaloes (50 primiparous and 50 pluriparous) of five popular breeds were recruited to characterize and compare teat morphology. In particular, the focus was put on the Nili Ravi, Mediterranean, Egyptian, Bulgarian Murrah, and Azeri buffaloes raised in Pakistan, Italy, Egypt, Bulgaria, and Iran, respectively. In all countries, a longitudinal cross-section ultrasound was obtained before the milking to measure teat parameters at individual level: overall, teat canal length (TCL) averaged 24.13 mm, teat diameter (TD) 30.46 mm, cisternal diameter (CD) 17.80 mm, and teat wall (TW) 7.12 mm. The most variable trait across breeds was TCL which was positively correlated with CD and TD and negatively with TW, regardless of the teat position (front/rear or left/right). A strong negative correlation was found between TW and CD (- 0.43). The analysis of variance revealed that the fixed effect of breed significantly affected all the traits except TD. In fact, Bulgarian Murrah, Azeri, and Egyptian buffaloes presented the greatest estimate of TCL, whereas NR the smallest (14.70 mm). The TW was maximum in Nili Ravi, Egyptian, and Mediterranean buffaloes, with estimates equal to 8.19, 7.59, and 8.74 mm, respectively. Nili Ravi also showed the greatest TL (82.39 mm). In terms of CD, the lowest least square mean was that of Mediterranean buffaloes (12.14 mm). Primiparous and pluriparous buffaloes differed in terms of TD, TW, and TL, with older animals presenting the highest least square mean. In terms of position, instead, significant differences were observed for TD, CD, and TL when comparing front and rear teats, as left and right teats did not differ. Teat anatomy includes a set of heritable morphological features and is therefore breed-dependent. Differences presented in this study could be attributed to the divergent breeding objective and selective pressure across the five breeds; e.g., in some cases such as Mediterranean buffalo, selection for decades was oriented to improve milk production and milkability and achieve optimal conformation for mechanical milking. A better understanding of the mammary gland anatomical descriptors can be informative of the history of a breed and could provide useful insights to guide possible selection.

Ultrasonographic, endoscopic and radiographic examinations of the dromedary mammary glands and teats.

The aim of the present study was to examine the mammary gland of dromedary camels using ultrasonography, endoscopy and radiography. These techniques are easy to perform in the field and feasible to diagnose pathological conditions of the mammary gland. Udders of 49 slaughtered and 26 adult dromedary camels submitted for necropsy were used for the examinations. Additionally, 11 lactating female dromedary camels were selected for the ultrasonographic udder examination. The transition from the milk ducts into the udder cistern, the teat cistern and the teat canals were examined in individual udders. Teat cistern length, teat end width, teat wall thickness, teat cistern width and middle cistern wall thickness were measured using ultrasonography. The measurements resulted in mean values of the teat cistern length of 37.3 mm, the teat end width of 2.0 mm, the teat wall thickness of 4.4 mm, the teat cistern width of 8.2 mm and the cistern wall thickness of 3.5 mm. The teat wall was differentiated into three layers, a hyperechoic outer layer, a hypoechoic middle layer and a hyperechoic inner layer. The mid cistern wall was hyperechoic. Endoscopic examination is an easy to perform and practicable method for examining the inner structures of the teats of dead animals; however, the feasibility has not been shown in lactating animals yet. Ring-like folds were present in the teat cistern, which protruded horizontally into the lumen. It was also possible to visualize the branchlike transition of the teat cistern into the larger milk ducts. Radiographic examination using barium sulfate contrast medium showed that the teat cistern ends in a network of initially wide but branching and narrowing milk ducts. The two teat canals and cisterns are completely independent of each other and there is no communication between the glandular tissue of the two canals and cisterns.

Longitudinal analysis of bovine mammary gland development.

Many studies on bovine mammary glands focus on one stage of development. Often missing in those studies are repeated measures of development from the same animals. As milk production is directly affected by amount of parenchymal tissue within the udder, understanding mammary gland growth along with visualization of its structures during development is essential. Therefore, analysis of ultrasound and histology data from the same animals would result in better understanding of mammary development over time. Thus, this research aimed to describe mammary gland development using non-invasive and invasive tools to delineate growth rate of glandular tissue responsible for potential future milk production. Mammary gland ultrasound images, biopsy samples, and blood samples were collected from 36 heifer dairy calves beginning at 10 weeks of age, and evaluated at 26, 39, and 52 weeks. Parenchyma was quantified at 10 weeks of age using ultrasound imaging and histological evaluation, and average echogenicity was utilized to quantify parenchyma at later stages of development. A significant negative correlation was detected between average echogenicity of parenchyma at 10 weeks and total adipose as a percent of histological whole tissue at 52 weeks. Additionally, a negative correlation between average daily gain at 10 and 26 weeks and maximum echogenicity at 52 weeks was present. These results suggest average daily gain and mammary gland development prior to 39 weeks of age is associated with development of the mammary gland after 39 weeks. These findings could be predictors of future milk production, however this must be further explored.

Mouse Mammary Gland Whole Mount Density Assessment across Different Morphologies Using a Bifurcated Program for Image Processing.

Mammographic density is associated with increased breast cancer risk. Conventional visual assessment of murine mouse models does not include quantified total density analysis. A bifurcated method was sufficient to obtain relative density scores on a broad range of two-dimensional whole mount images that contained both normal and abnormal findings. Image processing techniques, including a ridge operator and a gaussian denoising method, were used to isolate background away from mammary epithelium and use mean pixel intensity to represent mammary density on genetically engineered mouse models for breast cancer in mice 4 to 29 months of age. The bifurcated method allowed for application of an optimal image processing approach for the structural elements present in the whole mount images. Gaussian denoising was the optimal approach when more dense lobular growth and tertiary branching dominate and a ridge operator when epithelial growth was more sparse and secondary branching was the more dominant structural feature. The two processing approaches were combined in a single experimental flow program using an initial image density measurement as the decision point between the two approaches. Higher density was associated with lobular growth, tertiary branching, fibrotic stroma, and presence of cancer. The significance of the study is development of a readily accessible program for digital assessment of mammary gland whole mount density across a range of mammary gland morphologies.

Multiscale imaging of basal cell dynamics in the functionally mature mammary gland.

The mammary epithelium is indispensable for the continued survival of more than 5,000 mammalian species. For some, the volume of milk ejected in a single day exceeds their entire blood volume. Here, we unveil the spatiotemporal properties of physiological signals that orchestrate the ejection of milk from alveolar units and its passage along the mammary ductal network. Using quantitative, multidimensional imaging of mammary cell ensembles from GCaMP6 transgenic mice, we reveal how stimulus evoked Ca2+ oscillations couple to contractions in basal epithelial cells. Moreover, we show that Ca2+-dependent contractions generate the requisite force to physically deform the innermost layer of luminal cells, compelling them to discharge the fluid that they produced and housed. Through the collective action of thousands of these biological positive-displacement pumps, each linked to a contractile ductal network, milk begins its passage toward the dependent neonate, seconds after the command.

Relationship between blood calcium level and post-milking teat canal closure in dairy cows.

The teat canal-one of the primary defense mechanisms of the udder-ensures the milk flow during milking in bovines and prevents pathogens from entering the udder by forming a barrier through the elastic muscle and keratin layers tightly closing the surrounding area. The current study investigated the effects of blood calcium status on teat closure in cows after milking. The study covered 200 healthy teats, of which 100 were from normocalcemic (NC) cows and 100 were from subclinical hypocalcemic (SCH) cows. Teat canal length (TCL) and width (TCW) were measured with ultrasonography at 0-min pre-milking and 15- and 30-min post-milking. Cylindrically shaped teat canal volume (TCV) was calculated by deriving from TCL and TCW. Time-dependent changes in teat canal closure and their relationships with blood calcium levels were analyzed. The results showed that the calcium level did not affect TCL, TCW, and TCV (P > 0.05) during the 15-min post-milking period. However, TCL (P < 0.001), TCW (P < 0.05), and TCV (P < 0.001) were lower in NC cows than in SCH cows at 30-min post-milking. At 15-min post-milking, no correlation existed between the teat canal closure (ΔTCL, ΔTCW, and ΔTCV) and the blood calcium level, while significant correlations were available between the teat canal closure and the blood calcium level {ΔTCL (r: - 0.288, P < 0.001), ΔTCW (r: - 0.260, P < 0.001), ΔTCV (r: - 0.150, P < 0.05)} at 30-min post-milking. The current study concluded that the blood calcium status significantly impacts the teat canal closure in bovines, and calcium status should be meticulously monitored with the mastitis control program to apply necessary strategic steps.

Magnetic resonance angiography reveals increased arterial blood supply and tumorigenesis following high fat feeding in a mouse model of triple-negative breast cancer.

Breast cancer is the second most commonly diagnosed malignancy among women globally. Past MRI studies have linked a high animal fat diet (HAFD) to increased mammary cancer risk in the SV40Tag mouse model of triple-negative breast cancer. Here, serial MRI examines tumor progression and measures the arterial blood volume feeding mammary glands in low fat diet (LFD) or HAFD fed mice. Virgin female C3(1)SV40Tag mice (n = 8), weaned at 3 weeks old, were assigned to an LFD (n = 4, 3.7 kcal/g, 17.2% kcal from vegetable oil) or an HAFD (n = 4, 5.3 kcal/g, 60% kcal from lard) group. From ages 8 to 12 weeks, weekly fast spin echo MR images and time-of-flight (TOF) MR angiography of inguinal mammary glands were acquired at 9.4 T. Following in vivo MRI, mice were sacrificed. Inguinal mammary glands were excised and fixed for ex vivo MRI and histology. Tumor, blood, and mammary gland volumes for each time point were measured from manually traced regions of interest; tumors were classified as invasive by histopathology-blinded observers. Our analysis confirmed a strong correlation between total tumor volume and blood volume in the mammary gland. Tumor growth rates from weeks 8-12 were twice as high in HAFD-fed mice (0.42 ± 0.14/week) as in LFD-fed mice (0.21 ± 0.03/week), p < 0.004. Mammary gland blood volume growth rate was 2.2 times higher in HAFD mice (0.29 ± 0.11/week) compared with LFD mice (0.13 ± 0.06/week), p < 0.02. The mammary gland growth rate of HAFD-fed mice (0.071 ± 0.011/week) was 2.7 times larger than that of LFD-fed mice (0.026 ± 0.009/week), p < 0.01. This is the first non-invasive, in vivo MRI study to demonstrate a strong correlation between an HAFD and increased cancer burden and blood volume in mammary cancer without using contrast agents, strengthening the evidence supporting the adverse effects of an HAFD on mammary cancer. These results support the potential future use of TOF angiography to evaluate vasculature of suspicious lesions.

Different vacuum levels, vacuum reduction during low milk flow, and different cluster detachment levels affect milking performance and teat condition in dairy cows.

Traditionally, machine milking is performed at a constant vacuum supply. The system vacuum has to be set high enough to allow a sufficiently high vacuum at the teat end, despite the inevitable vacuum drop caused by milk flow. This leads to an increased vacuum load on the teat, especially when milk flow ceases at the end of milking. We tested the hypothesis that a milk flow-controlled adaptation of vacuum settings during milking allows even higher vacuum levels than are usually recommended during the period of high milk flow if the vacuum is reduced during low milk flow. Combined with a high cluster detachment flow rate level, increased milking performance is expected without an increased effect on teat tissue. Ten Holstein dairy cows were milked with a bucket milker with the claw vacuum adjusted in the absence of milk flow at a regular (43 kPa) and high (48 kPa) claw vacuum, with and without vacuum reduction during low milk flow (<2 kg/min), and combined with different cluster detachment levels (0.2, 0.6, and 1 kg/min). Each treatment was applied in each cow during 4 subsequent milkings in a randomized crossover design. Both claw vacuum and milk flow were continuously recorded throughout milking. Teat tissue thickness was measured using a cutimeter and teat wall diameter was measured by B-mode ultrasonography at 5 min after the end of milking. Milk yield was not affected by either vacuum settings or detachment levels. Machine-on time in treatments with vacuum reduction was shorter at high than at low vacuum and decreased with increasing detachment levels. Average milk flow was higher at high than at low vacuum and reached highest values in milkings without vacuum reduction at both vacuum levels. The average milk flow was higher at a cluster detachment of 1 kg/min than at 0.2 kg/min. However, both teat tissue thickness and (as a tendency) teat wall diameter at 5 min after cluster detachment were higher in milkings at high vacuum without vacuum reduction compared with all other treatments. In conclusion, high claw vacuum up to 48 kPa increases milking performance because of higher milk flow and reduced machine-on time. Negative effects of high vacuum on teat tissue are prevented by reducing vacuum during low milk flow (<2 kg/min) at the start and end of milking. Additionally, using a high cluster detachment level reduces machine-on time without a loss of harvested milk.

Short communication: Teat wall diameter and teat tissue thickness in dairy cows are affected by intramammary pressure and by the mechanical forces of machine milking.

The objective of this study was to validate and apply 2 different methods to record changes in teat tissue related to machine milking. Teat wall diameter was measured via B-mode ultrasound cross sectioning with a 7.5-MHz linear probe. Teat tissue thickness was measured using a cutimeter (spring-loaded caliper, spring constant 6.5 N/cm, 0.5 N at closed jaws). Both methods were applied at the teat barrel, 2 cm above the teat tip. In experiment 1, 24 teats from freshly slaughtered cows were used to perform ultrasound imaging (12 teats) or cutimeter measurements (12 teats) while the teat cisterns were filled with water to increase the intracisternal pressure from 0 to 30 kPa in steps of 1 kPa. Teat tissue thickness did not change at an intracisternal pressure from 0 to 10 kPa but increased with intracisternal pressure at levels >10 kPa. In contrast, teat wall diameter decreased with intracisternal pressure between 0 and 7 kPa but did not significantly change at a pressure ≥7 kPa up to 30 kPa. Significant Pearson correlation coefficients between intracisternal pressure and teat wall diameter were observed from 0 to 7 kPa (r = -0.38), and between intracisternal pressure and teat tissue thickness from 10 to 30 kPa (r = 0.45). In experiment 2, ultrasound and cutimeter measurements were performed in 12 lactating Holstein cows. Measurements before and during milking, immediately after cluster removal, with normal milking or with a 5-min overmilking, were performed and continued at 5-min intervals for 60 min and at 10-min intervals until 120 min. Additionally, with the 5-min overmilking treatment, measurements were continued at 60-min intervals up to 10 h after milking. Teat wall diameter decreased in response to milk ejection, followed by a continuous increase during the course of milking, with highest values after 5 min overmilking. Teat tissue thickness did not change during milking but was significantly increased after overmilking. Teat wall diameter and teat tissue thickness recovered to premilking levels within 35 min after normal milking and within 60 min after overmilking. Until 10 h after overmilking, the teat wall diameter decreased steadily, whereas teat tissue thickness was unfluctuating. In the physiologically relevant range of intramammary pressure, ultrasound measurements of the teat wall were affected by both intramammary pressure and mechanical forces, whereas cutimeter measurements were not affected by the intramammary pressure.

Blood perfusion of teat tissue in dairy cows: Changes associated with pre-milking stimulation and machine milking.

Mechanical forces during machine milking of dairy cows evoke circulatory impairment of the teat tissue that may affect the teats' defense mechanisms against mastitis pathogens. Ample research describes dimensional changes of different teat traits after machine milking, whereas reports that describe changes in blood circulation of dairy cows' teats are limited. Therefore, the objectives of this study were to (1) describe changes in teat blood circulation that occur after pre-milking teat stimulation and machine milking and (2) study the effect of 2 different milking liners on machine milking-induced changes in teat blood flow. In a randomized trial, Holstein dairy cows were stratified by parity, stage of lactation, and average daily milk yield during the previous week, and allocated to 1 of 2 treatment groups. Treatment consisted of 1 milking observation with either a round or multisided concave milking liner. Teat scans were taken of the left front and the right hind teats using power Doppler ultrasonography. Imaging occurred before pre-milking udder preparation (T1), after completion of pre-milking udder preparation but before milking-unit attachment (T2), and immediately after unit detachment (T3). Perfusion intensity measurements from teat scans were performed with a commercially available software program. Data from 109 cows were analyzed. A general linear mixed model showed differences in perfusion intensity between time points. Least squares means (95% confidence intervals) for T1, T2, and T3, respectively, were 0.035% (0.026-0.047), 0.124% (0.093-0.164), and 0.095% (0.073-0.124). Conversely, no statistically significant differences between treatment groups were observed. We conclude that teat blood circulation is subjected to several influences, including inherent circulatory regulation mechanisms, as well as extrinsic factors such as machine milking. Future research is warranted to decipher the magnitude of their influence and to further our understanding of how these changes relate to the susceptibility to intramammary infection and milking performance.

Ductal tree ablation by local delivery of ethanol prevents tumor formation in an aggressive mouse model of breast cancer.

BACKGROUND: Prophylactic mastectomy is the most effective intervention to prevent breast cancer. However, this major surgery has life-changing consequences at the physical, emotional, psychological, and social levels. Therefore, only high-risk individuals consider this aggressive procedure, which completely removes the mammary epithelial cells from which breast cancer arises along with surrounding tissue. Here, we seek to develop a minimally invasive procedure as an alternative to prophylactic mastectomy by intraductal (ID) delivery of a cell-killing solution that locally ablates the mammary epithelial cells before they become malignant. METHODS: After ID injection of a 70% ethanol-containing solution in FVB/NJ female animals, ex vivo dual stained whole-mount tissue analysis and in vivo X-ray microcomputed tomography imaging were used to visualize ductal tree filling, and histological and multiplex immunohistochemical assays were used to characterize ablative effects and quantitate the number of intact epithelial cells and stroma. After ID injection of 70% ethanol or other solutions in cancer-prone FVB-Tg-C3(1)-TAg female animals, mammary glands were palpated weekly to establish tumor latency and examined after necropsy to record tumor incidence. Statistical difference in median tumor latency and tumor incidence between experimental groups was analyzed by log-rank test and logistic mixed-effects model, respectively. RESULTS: We report that ID injection of 70% ethanol effectively ablates the mammary epithelia with limited collateral damage to surrounding stroma and vasculature in the murine ductal tree. ID injection of 70% ethanol into the mammary glands of the C3(1)-TAg multifocal breast cancer model significantly delayed tumor formation (median latency of 150 days in the untreated control group [n = 25] vs. 217 days in the ethanol-treated group [n = 13], p value < 0.0001) and reduced tumor incidence (34% of glands with tumors [85 of 250] in the untreated control group vs. 7.3% of glands with tumor [7 of 95] in the ethanol-treated group, risk ratio = 4.76 [95% CI 1.89 to 11.97, p value < 0.0001]). CONCLUSIONS: This preclinical study demonstrates the feasibility of local ductal tree ablation as a novel strategy for primary prevention of breast cancer. Given the existing clinical uses of ethanol, ethanol-based ablation protocols could be readily implemented in first-in-human clinical trials for high-risk individuals.

Local wavefront mapping in tissue using computational adaptive optics OCT.

The identification and correction of wavefront aberrations is often necessary to achieve high-resolution optical images of biological tissues, as imperfections in the optical system and the tissue itself distort the imaging beam. Measuring the localized wavefront aberration provides information on where the beam is distorted and how severely. We have recently developed a method to estimate the single-pass wavefront aberrations from complex optical coherence tomography (OCT) data. Using this method, localized wavefront measurement and correction using computational OCT was performed in ex vivo tissues. The computationally measured wavefront varied throughout the imaged OCT volumes and, therefore, a local wavefront correction outperformed a global wavefront correction. The local wavefront measurement was also used to generate tissue aberration maps. Such aberration maps could potentially be used as a new form of tissue contrast.

High-speed, low-cost, pulsed-laser-diode-based second-generation desktop photoacoustic tomography system.

Bulky, expensive Nd:YAG lasers are used in conventional photoacoustic tomography (PAT) systems, making them difficult to translate into clinics. Moreover, real-time imaging is not feasible when a single-element ultrasound transducer is used with these low-pulse-repetition-rate lasers (10-100 Hz). Low-cost pulsed laser diodes (PLDs) can be used instead for photoacoustic imaging due to their high-pulse-repetition rates and compact size. Together with acoustic-reflector-based multiple single-element ultrasound transducers, a portable desktop PAT system was developed. This second-generation PLD-based PAT achieved 0.5 s cross-sectional imaging time with high spatial resolution of ∼165 µm and an imaging depth of 3 cm. The performance of this system was characterized using phantom and in vivo studies. Dynamic in vivo imaging was also demonstrated by monitoring the fast uptake and clearance of indocyanine green in small animal (rat) brain vasculature.

Development and Validation of an Image Analysis System for the Measurement of Cell Proliferation in Mammary Glands of Rats.

Reliable detection and measurement of cell proliferation are essential in the preclinical assessment of carcinogenic risk of therapeutics. In this context, the assessment of mitogenic potential on mammary glands is crucial in the preclinical safety evaluation of novel insulins. The existing manual counting is time-consuming and subject to operator bias. To standardize the processes, make it faster, and resistant to errors, we developed a semiautomated image analysis system (CEPA software, which is open-source) for counting of proliferating cells in photomicrographs of mammary gland sections of rats labeled with Ki-67. We validated the software and met the predefined targets for specificity, accuracy, and reproducibility. In comparison to manual counting, the respective mean differences in absolute labeling indices (LIs) for CEPA software were 3.12% for user 1 and 3.05% for user 2. The respective regression analysis revealed a good correlation between the CEPA software user and manual counting. Moreover, the CEPA software showed enhanced reproducibility between independent users. The interuser variability is centered around 0 and the absolute difference was about 0.53% LI. Based on validation data, our software has superiority to the manual counting and is a valid and reliable tool for the routine analysis of cell proliferation in mammary glands from rats exposed to insulin analogs.

Machine milking-induced changes in teat canal dimensions as assessed by ultrasonography.

Changes in teat canal diameter as induced by machine milking are thought to be related to teat canal penetrability and susceptibility to new intramammary infections. The objective of this study was to describe the effects of machine milking on teat canal dimensions as assessed by ultrasonography and to evaluate postmilking changes of teat canal dimensions throughout an 8-h milking interval. Ultrasonographic images were taken of the left front and right hind teats of 80 Holstein cows milked 3 times per day. Imaging occurred before attachment of the milking unit after completion of premilking udder preparation (t-1), immediately after removal of the milking unit (t0), and then every hour after milking until 8 h had elapsed (t1 to t8). Teat canal length and teat canal diameter (at the proximal, middle, and distal regions) were measured. General linear mixed model analysis indicated differences in the relative change of teat canal length (compared with t-1) with least squares means (LSM) and 95% confidence interval (95% CI) of 11.1% (8.7-13.4%) after machine milking (t0). Teat canal length decreased throughout the 8-h milking interval but remained elevated relative to t-1. Teat canal diameter at the proximal and middle regions increased near the end of the milking interval, whereas that at the distal region did not change meaningfully during the 8 h of observation. Our findings suggest that the teat canal and its surrounding tissue is in a state of near-constant remodeling in cows milked 3 times daily. Future work is warranted to study the association between ultrasonographically assessed changes of teat canal dimensions after machine milking and teat canal penetrability to mastitis-causing pathogens.

Technical note: Automatic evaluation of infrared thermal images by computerized active shape modeling of bovine udders challenged with Escherichia coli.

Mastitis causes substantial economic losses and animal suffering in the dairy industry. The trend toward larger herd sizes complicates the monitoring of udder health in individual animals. Infrared thermography has successfully been used for early mastitis detection. However, manual thermogram analysis is time consuming and requires a skilled examiner, and automated image processing has not been tested. The aim of this study was to determine whether automatic evaluation of thermograms showed results comparable to those of manual evaluation of thermograms. Five healthy cows underwent an intramammary challenge with Escherichia coli to induce clinical mastitis. Multiple udder thermograms were taken every 2 h for 24 h before and after the challenge, resulting in 4,143 images in total. All images were evaluated using image recognition software (automatically) and a polygon tool (manually) to calculate the average and maximum surface temperatures. Because of the slightly different regions of interest, temperatures ascertained from the thermograms using the automatic method were consistently lower than those ascertained using the manual method. However, average udder surface temperatures evaluated using both methods were strongly correlated (r = 0.98 in the left hindquarter, and r = 0.99 in the right hindquarter) and showed maximum temperature peaks at the same time, 13 and 15 h after intramammary challenge. In the receiver operating characteristic analysis, both methods provided good results for sensitivity and specificity in detecting clinical E. coli-induced mastitis at different threshold values. For automatically evaluated maximum right hindquarter temperature, sensitivity was 93.75% and specificity was 94.96%, and for manually evaluated maximum right hindquarter temperature, sensitivity was 93.75% and specificity was 96.40%. Thus, automatic thermogram evaluation is a promising tool for automated mastitis detection.

Technical note: Development and evaluation of a standardized technique to assess blood perfusion in teats of dairy cows using power Doppler ultrasonography.

Ample research has described the assessment of dimensional changes for different teat traits, whereas diagnostic techniques to reliably assess blood circulation in teats of dairy cows are limited. Here, we describe the development and evaluation of a scanning technique to quantify blood flow in teats of dairy cows using power Doppler ultrasonography. In 2 consecutive trials, 384 teat scans [trial 1, n = 256 (sagittal plane, n = 128; transverse plane, n = 128); trial 2, n = 128 (transverse plane)] from 16 cows were obtained by the same 2 operators. Perfusion intensity from single images (trial 1) and video images (trial 2) were assessed using a commercially available software program. Intraclass correlation coefficients (ICC) and concordance correlation coefficients (CCC) were used to assess interoperator reproducibility (agreement between measurements performed by different operators) and intraoperator repeatability (agreement between measurements performed by the same operator). In trial 1, interoperator ICC and CCC indicated poor agreement (ICC ≤0.26, CCC ≤0.26). Intraoperator ICC and CCC demonstrated poor agreement between duplicate measurements within operators (ICC ≤0.19, CCC ≤0.19). Modifications after trial 1 included (1) a different ultrasound device, (2) analysis of video clips rather than single images, (3) restriction to 1 sectional plane (i.e., transverse), and (4) a scanning sequence such that repeated scans within operators were measured one after another. Through these modifications, intraoperator repeatability in trial 2 yielded fair to good agreement, with intraoperator ICC and CCC over both operators ranging from 0.44 to 0.70 and from 0.57 to 0.69, respectively, whereas interoperator ICC and CCC showed poor agreement (ICC = 0.35, CCC = 0.34). We conclude that repeatable measurements of blood perfusion intensity of teats in dairy cows can be attained with power Doppler ultrasonography. Power Doppler ultrasonography is a suitable tool to quantify slow flow in small vessels and may be an acceptable diagnostic technique to assess changes in blood circulation that result from machine milking in teats of dairy cows, although further research is necessary to validate this hypothesis.

Short communication: Risk factors for sonographically detectable udder edema in overbagged cows at dairy shows.

At dairy cow shows, the appearance of the udder is very important. To show cows at their best, udders are often presented in an overbagged state by extending milking intervals before the show. This practice represents an animal welfare issue because it is associated with behavior changes (e.g., excessive abduction and decreased eating time) and may affect the health of the animal. The aim of the current study was to determine the association between various parameters (e.g., milk yield, days in milk, milking procedures, or administration of nonsteroidal anti-inflammatory drugs, steroids, or diuretics) and the sonographically detectable udder edema score. Data were collected from the show catalogs, by interviewing the exhibitors, by analyzing official veterinary treatment protocols and official milking data, via laboratory analysis of blood samples, and by sonographic examinations of the udders at the show. For sonographically detectable udder edema scoring, 3 scans were taken, 1 on each fore quarter and 1 incorporating both rear quarters at the level of the median suspensory ligament. For grading the scans, a scoring system of 4 grades (score 0 = no edema, 1 = slight edema, 2 = moderate edema, 3 = severe edema) was used. Data from 321 cows of different breeds were collected at 4 highly competitive Swiss dairy shows (shows A-D) between January and September 2017. To determine risk factors for sonographically detectable udder edema, data were analyzed with 2 different logistic regression models. In model 1, the odds ratios were 3.33 (milking intervals of ≥16 h vs. <14 h), 3.84 (show A vs. show C), and 7.39 (dairy breeds vs. dual-purpose breeds). Milking intervals of dairy breeds were significantly higher than those of dual-purpose breeds. In model 2, milking interval was the only significant risk factor with an odds ratio of 9.00 for milking intervals of ≥16 h. Only the milking intervals represented a relevant risk factor in both models; therefore, we concluded that the previously described sonographic udder edema scoring is a useful technique for detecting overbagged udders at dairy shows. Its routine implementation may improve cow welfare at dairy shows.

The relationship between mastitis and the B-mode, colour Doppler ultrasonography measurements of supramammary lymph nodes in cows.

This research communication aims to test the hypothesis that B-Mode, colour Doppler ultrasonographic measurements and characteristics can identify mastitis in dairy cows. A total of 102 lactating cows were divided into 3 groups: cows in which all mammary quarters were CMT-negative, cows with CMT-positive mammary quarters and cows with clinical mastitis in at least one quarter. Colour Doppler ultrasonography measurements of the supramammary lymph nodes revealed that distortion-type vascular morphology, the rate of type 4 vascular densities and the incidence of mixed-type vascular distributions were highest in the clinical mastitis group, whereas the frequency of avascularity in supramammary lymph nodes was highest in the CMT-negative group. All differences were significant. In conclusion, the use of B-mode and colour Doppler ultrasonographic measurements of the supramammary lymph nodes can provide useful information about the current condition of mastitis in cows, although its diagnostic potential remains to be determined.