Impact Evaluation of Score Classes and Annotation Regions in Deep Learning-Based Dairy Cow Body Condition Prediction.

Body condition scoring is a simple method to estimate the energy supply of dairy cattle. Our study aims to investigate the accuracy with which supervised machine learning, specifically a deep convolutional neural network (CNN), can be used to retrieve body condition score (BCS) classes estimated by an expert. We recorded images of animals' rumps in three large-scale farms using a simple action camera. The images were annotated with classes and three different-sized bounding boxes by an expert. A CNN pretrained model was fine-tuned on 12 and 3 BCS classes. Training in 12 classes with a 0 error range, the Cohen's kappa value yielded minimal agreement between the model predictions and ground truth. Allowing an error range of 0.25, we obtained minimum or weak agreement. With an error range of 0.5, we had strong or almost perfect agreement. The kappa values for the approach trained on three classes show that we can classify all animals into BCS categories with at least moderate agreement. Furthermore, CNNs trained on 3 BCS classes showed a remarkably higher proportion of strong agreement than those trained in 12 classes. The prediction precision when training with various annotation region sizes showed no meaningful differences. The weights of our trained CNNs are freely available, supporting similar works.

Isocyanide Substitution in Acridine Orange Shifts DNA Damage-Mediated Phototoxicity to Permeabilization of the Lysosomal Membrane in Cancer Cells.

In cancer therapy, immunogenic cell death eliminates tumor cells more efficiently than conventional apoptosis. During photodynamic therapy (PDT), some photosensitizer (PS) targeting lysosomes divert apoptosis to the immunologically more relevant necrosis-like cell death. Acridine orange (AO) is a PS targeting lysosome. We synthesized a new compound, 3-N,N-dimethylamino-6-isocyanoacridine (DM), a modified AO, aiming to target lysosomes better. To compare DM and AO, we studied optical properties, toxicity, cell internalization, and phototoxicity. In addition, light-mediated effects were monitored by the recently developed QUINESIn method on nuclei, and membrane stability, morphology, and function of lysosomes utilizing fluorescent probes by imaging cytometry in single cells. DM proved to be a better lysosomal marker at 405 nm excitation and lysed lysosomes more efficiently. AO injured DNA and histones more extensively than DM. Remarkably, DM's optical properties helped visualize shockwaves of nuclear DNA released from cells during the PDT. The asymmetric polar modification of the AO leads to a new compound, DM, which has increased efficacy in targeting and disrupting lysosomes. Suitable AO modification may boost adaptive immune response making PDT more efficient.

Serological screening for Coxiella burnetii in the context of early pregnancy loss in dairy cows.

Q fever is one of the commonest infectious diseases worldwide. A Coxiella burnetii prevalence of 97.6% has been found by ELISA and PCR tests of the bulk tank milk in dairy cattle farms of Hungary. The herd- and individual-level seroprevalence rates of C. burnetii in the examined dairy cows and farms have dramatically increased over the past ten years. Three high-producing industrial dairy farms were studied which had previously been found ELISA and PCR positive for C. burnetii by bulk tank milk testing. Coxiella burnetii was detected in 52% of the 321 cows tested by ELISA. Pregnancy loss was detected in 18% of the cows between days 29-35 and days 60-70 of gestation. The study found a higher seropositivity rate (80.5%) in the cows that had lost their pregnancy and a seropositivity of 94.4% in the first-bred cows that had lost their pregnancy at an early stage. The ELISA-positive pregnant and aborted cows were further investigated by the complement fixation test (CFT). In dairy herds an average of 66.6% individual seropositivity was detected by the CFT (Phase II) in previously ELISA-positive animals that had lost their pregnancy and 64.5% in the pregnant animals. A higher (Phase I) seropositivity rate (50.0%) was found in the cows with pregnancy loss than in the pregnant animals (38.5%). The high prevalence of C. burnetii in dairy farms is a major risk factor related to pregnancy loss.

Relationship between reproductive management practices and fertility in primiparous and multiparous dairy cows.

The aim of this study was to analyze the impact of reproductive management practices on fertility in primiparous and multiparous dairy cows and to identify the differences by parity. Data on reproductive performance and management of cows were gathered from 34 large commercial Holstein-Friesian dairy herds in Hungary. Individual data on 23 781 cows that calved in the studied herds in 2014 were collected from the national milk recording database and used to calculate the reproductive indices. Farm managers and veterinarians were interviewed in person from May 22 to November 6, 2015 using a questionnaire to collect information about reproductive management practices. The data were statistically analyzed by mixed-effects models. Our results showed that the use of a voluntary waiting period (VWP) was linked to a greater increase in calving-to-conception interval (CCI, P < 0.05) and a greater decline in the chance of pregnancy at 200 days in milk (P200, P < 0.001) in multiparous cows. Estrus synchronization was related to a larger reduction in days-to-first service (DFS, P < 0.001), breeding interval (IBI, P < 0.05), and CCI (P < 0.01) and a greater improvement in P200 (P < 0.001) in primiparous cows. Early pregnancy diagnosis and pregnancy recheck improved IBI (P < 0.01 for both practices), CCI (P < 0.01 and P < 0.001, respectively), and P200 (P < 0.001 for both practices) to a larger extent in primiparous than in multiparous cows. In conclusion, estrus synchronization, early pregnancy diagnosis, and pregnancy recheck led to a greater improvement in fertility in primiparous cows than in multiparous cows.

L'objectif de la présente étude était d'analyser l'impact des pratiques de gestion de la reproduction sur la fertilité de vaches laitières primipares et multipares et d'identifier les différences par parité. Les données sur les performances de reproduction et la gestion des vaches ont été amassées auprès de 34 fermes laitières Holstein-Friesian commerciales importantes en Hongrie. Les données individuelles de 23 781 vaches ayant mise-bas en 2014 dans les troupeaux à l'étude ont été obtenues de la base de données nationale d'enregistrement de la production laitière et furent utilisées pour calculer les indices de reproduction. Les gérants de ferme et les vétérinaires ont été interviewés en personne entre le 22 mai et le 6 novembre 2015 à l'aide d'un questionnaire pour obtenir des informations sur les pratiques de gestion de la reproduction. Les données ont été analysées statistiquement par des modèles à effets mixtes. Nos résultats ont démontré que l'utilisation d'une période volontaire d'attente (VWP) était liée à une plus grande augmentation de l'intervalle vêlage-conception (CCI, P < 0,05) et à un plus grand déclin de la probabilité de gestation à 200 jours en lait (P200, P < 0,001) chez les vaches multipares. La synchronisation des oestrus était reliée à une plus grande réduction du nombre de jours avant la première saillie (DFS, P < 0,001), de l'intervalle de reproduction (P < 0,05), et de CCI (P < 0,01), et une meilleure amélioration du P200 (P < 0,001) chez les vaches primipares. Des diagnostics de gestation hâtifs et des vérifications de la gestation ont amélioré IBI (P < 0,01 pour ces deux pratiques), CCI (P < 0,01 et P < 0,001, respectivement), et P200 (P < 0,001 pour les deux pratiques) d'une manière plus importante chez les primipares que chez les multipares. En conclusion, la synchronisation des oestrus, les diagnostics de gestation hâtif, et la vérification de la gestation ont conduit à une plus grande amélioration de la fertilité chez les vaches primipares que chez les vaches pluripares.(Traduit par Docteur Serge Messier).

Effect of single-nucleotide polymorphisms on specific reproduction parameters in Hungarian Large White sows.

The aim of this study was to reveal the effect of single-nucleotide polymorphisms (SNPs) on the total number of piglets born (TNB), the litter weight born alive (LWA), the number of piglets born dead (NBD), the average litter weight on the 21st day (M21D) and the interval between litters (IBL). Genotypes were determined on a high-density Illumina Porcine SNP 60K BeadChip. Data screening and data identification were performed by a multi-locus mixed-model. Statistical analyses were carried out to find associations between individual genotypes of 290 Hungarian Large White sows and the investigated reproduction parameters. According to the analysis outcome, three SNPs were identified to be associated with TNB. These loci are located on chromosomes 1, 6 and 13 (-log10P = 6.0, 7.86 and 6.22, the frequencies of their minor alleles, MAF, were 0.298, 0.299 and 0.364, respectively). Two loci showed considerable association (-log10P = 10.35 and 10.46) with LWA on chromosomes 5 and X, the MAF were 0.425 and 0.446, respectively. Seven loci were found to be associated with NBD. These loci are located on chromosomes 5, 6, 13, 14, 15, 16 and 18 (-log10P = 10.95, 5.43, 8.29, 6.72, 6.81, 5.90, and 5.15, respectively). One locus showed association (-log10P = 5.62) with M21D on chromosome 1 (the MAF was 0.461). Another locus was found to be associated with IBL on chromosome 8 (-log10P = 7.56; the MAF was 0.438). The above-mentioned loci provide a straightforward possibility to assist selection by molecular tools and, consequently, to improve the competitiveness of the Hungarian Large White (HLW) breed.

Luteal activity following follicular drainage of subordinate follicles for twin pregnancy prevention in bi-ovular dairy cows.

Twin pregnancy is undesirable in dairy cattle. This study examines luteal activity following ultrasound-guided puncture and drainage of the smaller pre-ovulatory follicle at timed AI in cows with a pre-ovulatory follicle in each ovary. Luteal activity was determined through Doppler ultrasonography and plasma progesterone (P4) concentrations. The effects of GnRH treatment on Day 7 post-AI on subsequent luteal activity were also assessed. Two study groups were established: a control group of 29 cows and a follicular drainage (FD) group of 28 cows. After drainage, all cows developed a corpus luteum (CL) in the drained ovary. On Day 21 post-AI, drainage-induced CL and fellow CL were similar in terms of size and vascularization. According to a GLM repeated measures analysis of variance (P < 0.001), non-treated drained cows had lower P4 concentrations on Day 21 post-AI than non-treated non-drained cows, whereas GnRH treated cows, both drained and non-drained, showed the highest P4 concentrations at this time point. Twin pregnancy was recorded in 3 of the 8 pregnant control cows, whereas no twins were observed in the FD group. Our results indicate that luteal structures following follicular drainage were functional. As for the presence of an additional CL, this could suggest a reduced risk of pregnancy loss. In addition, luteal activity was significantly increased following GnRH treatment on Day 7 post-AI in drained cows.

Is twin pregnancy, calving and pregnancy loss predictable by serum pregnancy-specific protein b (pspb) concentration 28-35 days after ai in dairy cows?

Double ovulation occurs more frequently in multiparous cows with high milk production than in primiparous cows and the rate of twin pregnancy/calving is increasing worldwide. Diagnosis of twin pregnancy is possible by ultrasound at the time of early pregnancy examination [28-34 days after artificial insemination (AI)]. Pregnancy proteins are also well-known indicators of gestation. The risk of pregnancy loss during the first trimester of gestation for cows carrying twins is three to nine times higher than for cows carrying singletons. Pregnancy-specific protein B (PSPB) is a good indicator not only of pregnancy but also of pregnancy loss. The aims of this study were (a) to collect calving data in some Hungarian Holstein-Friesian herds (n = 7,300) to compare PSPB serum concentrations (measured 29-35 days post insemination) in twin- and singleton-calving cows (Trial 1), and (b) to check the predictive value of PSPB serum concentration for twin pregnancy and pregnancy loss in high-producing Spanish Holstein-Friesian cows (n = 98; Trial 2). Our results showed almost 7% twin calving rate. Although hormonal treatments are commonly believed to be major causes of twin pregnancies, our data do not support this hypothesis. The only exception is the single PGF injection, which significantly increased twin calving. No effect of milk production on the risk of twin pregnancy was found, and twin pregnancy increased with parity. The AI bull, the bull's sire, the bull's grandfather and the cow's father also affected twin calving (P ≤ 0.02). We found much higher frequency of twin calving in cows diagnosed pregnant with higher than 3 ng/ml serum PSPB concentrations at 29-35 days after insemination. In Trial 2, non-significant but well-marked differences were found in PSPB serum concentration between singleton- and twin-pregnant cow samples (2.1 and 2.9 ng/ml) at different bleeding times. Probably the small size of the study population and the effects of milk production on PSPB values may explain this lack of significance.

Effect of single-nucleotide polymorphisms on the breeding value of fertility and breeding value of beef in Hungarian Simmental cattle.

The objective of this study was to estimate the effect of single-nucleotide polymorphisms (SNPs) on the breeding value of fertility (BVF) and the breeding value of beef (BVB) in Hungarian Simmental cattle. Genotypes were determined on a high-density Illumina Bovine DNA Chip. Data screening and data identification were performed by multi-locus mixed-model. Statistical analyses were carried out to find associations between individual genotypes and the investigated quality values. Three loci showed considerable association with BVF (-log10 P = 9.5, 9.9 and 14.5, respectively) on chromosomes 9, 28 and 29, respectively. The frequencies of their minor alleles (MAF) were 0.375, 0.355 and 0.354, respectively. Two loci showed association with BVB (-log10 P = 25.3 and 22.7) on chromosomes 2 and 11, respectively (their MAF were 0.438 and 0.229). The abovementioned loci provide a straightforward possibility to assist selection by molecular tools.

Detection of pregnancy in sheep using an ELISA for pregnancy-specific protein B.

The early detection of pregnancy and the determination of fetal numbers have economic benefits in sheep production because of the seasonal breeding patterns where missing a breeding opportunity means the loss of one productive year. The purpose of this study was to evaluate the efficacy of the B6-HRP ELISA for ovine pregnancy-specific protein B (oPSPB) measurement in the detection of pregnancy and estimation of fetal numbers in different sheep breeds. BioPRYN® ELISA assay kit was used for the detection of pregnancy in the experimental animals. Ninety-three ewes of three breeds (British Milksheep - BM, Lacaune - L and Transylvanian Racka - TR), each from three farms in Hungary, were included in the study. BM and L ewes were artificially inseminated (AI). Thirty-five days after AI, all ewes were examined by transabdominal ultrasound. The TR flock was mated naturally over a six-week period. At the end of the mating period, the ewes were similarly examined by ultrasound. Blood samples were taken from all pregnant ewes twice (35 and 65 days after AI), and serum samples were assayed by the BioPRYN test. It can be concluded that the detection of serum PSPB by ELISA is a much easier, safer, less expensive and highly accurate method for the detection of ovine pregnancy. Although some breed-related differences were detectable at 35 and 65 days post breeding, no differences in oPSPB levels were found in pregnant ewes carrying different numbers of fetuses.

Macroscopic and histological characteristics of fluid-filled ovarian structures in dairy cows.

The primary objective of this study was to use macroscopic and histological features of corpora lutea with a cavity and anovulatory cystic ovarian structures, present in 90 pairs of abattoir-derived dairy cow ovaries, as the basis to clarify the nomenclature of ovarian structures. Excluding morphologically normal ovarian fol-licles (antrum < 2 cm, wall < 1 mm), there were 27 fluid-filled ovarian structures. Ovulatory structures > 16 mm in diameter were designated as Group A (cavity ≤ 10 mm and wall > 10 mm) or Group B (cavity > 10 mm and wall < 10 mm). The volume of luteal tissue was less (P < 0.05) in Group B than in Group A, whereas that of a solid corpus luteum (CL) was intermediate (least square means ± SEM: 72 ± 1.92, 11.22 ± 1.57 and 5.84 ± 1.92 cm3, respectively). There was a greater proportion (P < 0.05) of small luteal cells in Group B compared to a solid CL, whereas Group A was intermediate (58.6 ± 5.3, 37.4 ± 5.3 and 44.0 ± 4.4%, respectively). Connective tissue was thicker (P < 0.05) in Group B than in Group A (295.4 ± 46.9 vs. 153.9 ± 38.2 µm). Based on the above-mentioned characteristics and differences, Groups A and B were designated as a CL with a cavity and a cystic CL, respectively. Furthermore, there were three groups of anovulatory ovarian structures. Structures in Group C were termed persistent/anovulatory follicles (overall diameter and wall thickness ≤ 20 and 1-3 mm, respectively). Finally, Groups D and E were designated as a follicle-fibrous cyst and a follicle-luteinised cyst (based on histological structure) for anovulatory structures with an overall diameter and wall thickness of ≥ 20 and ≤ 3 mm, and ≥ 20 and ≥ 3 mm, respectively.

Estrus response and fertility after a single cloprostenol treatment in dairy cows with various ovarian structures.

The objective of this study was to determine rates of estrus and conception in lactating multiparous Holstein cows given 500 µg of cloprostenol intramuscularly after detection of the following ≥ 60 d after parturition: a solid corpus luteum (CL), a CL with a nonechodense cavity ≤ 20 mm in diameter (CLcav), a luteal cyst (cavity > 20 mm in diameter and a luteinized wall > 3 mm in diameter), or a follicular cyst (cavity > 20 mm and a luteinized wall ≤ 3 mm in diameter). The estrus rates were 335/419 (80.0%), 183/223 (82.1%), 170/182 (93.4%), and 44/87 (50.6%), respectively (P < 0.0001), and the conception rates 30 to 36 d after insemination among the estrous cows with an apparently normal mucus discharge were 130/285 (45.6%), 44/141 (31.2%), 39/79 (49.4%), and 19/30 (63.3%), respectively (P < 0.002). Compared with a solid CL, a CLcav did not affect the estrus rate but significantly reduced the conception rate (P < 0.05), and the estrus rates were significantly higher and lower in cows with a luteal or follicular cyst, respectively (P < 0.05).

L'objectif de la présente étude était de déterminer les taux d'oestrus et de conception chez des vaches Holstein multipares en lactation à qui on administra 500 µg de cloprosténol par voie intramusculaire après détection d'un des éléments suivants ≥ 60 j après la parturition : un corps jaune solide (CL), un CL avec une cavité non-échodense ≤ 20 mm de diamètre (CLcav), un kyste lutéal (cavité > 20 mm de diamètre et une paroi lutéinisée > 3 mm de diamètre), ou un kyste folliculaire (cavité > 20 mm et une paroi lutéinisée ≤ 3 mm de diamètre). Les taux d'oestrus étaient de 335/419 (80,0 %), 183/223 (82,1 %), 170/182 (93,4 %), et 44/87 (50,6 %), respectivement (P < 0,0001), et les taux de conception 30 à 36 j après insémination parmi les vaches en oestrus avec un écoulement de mucus d'apparence normale étaient 130/285 (45,6 %), 44/141 (31,2 %), 39/79 (49,4 %), et 19/30 (63,3 %), respectivement (P < 0,002). Comparativement à un CL solide, un CLcav n'a pas affecté le taux d'oestrus mais a réduit significativement le taux de conception (P < 0,05), et les taux d'oestrus étaient significativement plus élevés et plus faibles chez les vaches avec, respectivement, un kyste lutéal ou folliculaire (P < 0,05).(Traduit par Docteur Serge Messier).

Imaging of the parafoveal capillary network in diabetes.

The retinal vasculature is an extremely complex system that is adapted to support the metabolic demands of the retinal structures, but on the other hand maintain the optimal optical qualities of this tissue. Through histological studies and clinical studies using fluorescein angiography we have learned a lot about the retinal vasculature in its physiological state and in different diseases, but both of these study methods have serious limitations that limit their extensive application in healthy subjects or in patients with early disease. In this current review we will present early observations about the retinal vasculature from several novel noninvasive imaging modalities like adaptive optics SLO, retinal functional imager, adaptive optics OCT and Doppler OCT. Some of these instruments allow a more detailed in vivo examination of the retinal vasculature than fluorescein angiography without its potentially serious side effects, thus better allowing us to further study retinal vascular homeostasis in healthy subjects and to identify preclinical changes in early disease stages.

Photoreceptor layer regeneration is detectable in the human retina imaged by SD-OCT after laser treatment using subthreshold laser power.

PURPOSE: We evaluated the morphologic changes in retinal structure after laser photocoagulation using supra- and subthreshold laser fluence. METHODS: In a prospective cohort study 10 consecutive patients received scatter laser photocoagulation. Treatment was performed using a semiautomated patterned scanning laser system. In a study area adjacent to the temporal vessel arcades, 2 × 2 pattern laser spots were applied with halving the flux of the laser power in a stepwise manner starting from a power producing a typical grayish lesion. The study areas then were imaged on days one, three, and seven, and on months one, two, three, and six using color fundus photography, autofluorescence (AF), infrared (IR) imaging, and spectral domain optical coherence tomography (SD-OCT). RESULTS: The starting threshold power lesions each were visible on color fundus photography, IR, and AF in all patients, and showed characteristic changes on OCT throughout the follow-up period. The halved flux laser burns (first step) were undetectable ophthalmoscopically during the laser session, but during the follow-up always were detectable on IR and AF images, and sometimes on fundus photography. On OCT they showed changes similar to the suprathreshold laser scars, but were much smaller in diameter, and in some instances an inward migration of the photoreceptor layer was observed. CONCLUSIONS: Subthreshold laser burns with halved energy flux produced similar morphologic changes in the retina as threshold power, but with a smaller size. They induced less collateral damage to the neuroretina, and permit a level of reorganization in the outer retina. (ClinicalTrials.gov number, NCT00682240.).

Seasonal fertility differences in synchronised dairy cows: ultrasonic, metabolic and endocrine findings.

The aim of this study was to identify relationships among seasonal differences of fertility, metabolic parameters and appearance of irregular luteal forms in high-yielding dairy cows. Holstein-Friesian cows were put on the Provsynch regimen in winter (n = 10) and in summer (n = 10). Blood sampling (starting 35 days post partum) and rectal ultrasound examinations (starting post insemination) were carried out once a week in each examination period. Metabolic [plasma nonesterified fatty acid (NEFA) and beta-hydroxybutyrate (BHB) levels, ferric reducing ability of plasma (FRAP) and serum beta-carotene] and endocrine parameters [plasma thyroxine (T4), triiodothyronine (T3), insulin-like growth factor (IGF-I) and insulin levels] were measured. In summer, two cows were excluded from the study because of metritis and none of the remaining animals became pregnant, but 6 of the 8 cows had irregular luteal forms (ILF) on their ovaries. In winter, one cow was excluded because of metritis and 6 of the 9 cows became pregnant, while 2 of the 3 open cows had irregular luteal forms. In summer the mean plasma NEFA and BHB concentrations were significantly higher, while serum carotene and plasma IGF-I concentrations were significantly lower than in winter. The high plasma NEFA concentration found in summer seemed to be in association with the lower body condition score (BCS) caused by depressed appetite. In conclusion, statistical analysis supports the hypothesis that increased plasma NEFA and BHB and decreased plasma IGF-I concentrations may result in reduced fertility in summer. These changes may be associated with the more frequent appearance of ILFs and probably have a negative effect on ovarian function and/or oocyte quality.

A systematic correlation between morphology and functional alterations in diabetic macular edema.

PURPOSE: The aim of this study was to correlate different types of retinal morphologic alterations secondary to diabetic macular disease with their characteristic impact on retinal function. METHODS: In the present cross-sectional study, 26 eyes of 26 diabetic patients with clinically significant macular edema were examined. All patients underwent complete standardized ophthalmologic examination, including SD-OCT and microperimetry. Microperimetric values were projected over the scanning laser ophthalmoscope image of the OCT device, allowing direct correlation of functional and morphologic parameters. Results over all 1066 individual areas were analyzed using a general linear model. RESULTS: All the characteristic morphologic alterations demonstrated a significant effect on retinal function (P < 0.0002), except for outer nuclear layer (ONL) hyporeflectivity and small ONL cysts. Large ONL cysts (>220 µm) and serous retinal detachment had the greatest estimated negative effect on retinal sensitivity (-3.86 and -3.66 dB), followed by medium-sized ONL cysts, hard exudates associated with an extinction of the scan signal, and inner nuclear layer cysts. CONCLUSIONS: In diabetic macular edema, serous retinal detachment and large ONL cysts are the two morphologic changes with the greatest negative impact on retinal function.