Proliferation and apoptosis dynamics of the normal canine mammary gland during the oestrous cycle evaluated by Ki-67 and Caspase-3 indexes.

Understanding the normal physiology of the canine mammary gland (CMG) is crucial, as it provides a foundational reference for understanding canine mammary neoplasms. The relation between the Proliferation Index (PI) indicated by Ki-67 expression, along with the Apoptotic Index (AI) determined through Caspase-3 expression during the oestrous cycle, is inadequately documented in existing literature. This study seeks to offer insights into the interplay between PI and AI in the CMG across oestrous cycle phases. An extensive investigation was conducted on a diverse case series of bitches (n = 18). Oestrous cycle stages were determined through vaginal cytology, histological examination of the reproductive tract and serum progesterone and oestradiol concentrations. The entire mammary chain was histologically examined, and proliferation and apoptosis were assessed via double immunohistochemistry employing anti-Ki-67 and Caspase-3 antibodies. PI and AI were evaluated through a systematic random sampling approach, counting a minimum of 200 cells for each cell type. There was a significantly higher PI during early dioestrus in all mammary gland components, with a greater proportion of positive cells observed in epithelial cells compared to stromal cells. The highest PI was detected in epithelial cells within the end buds. Significant differences were found in Ki-67 labelling across the cranial mammary glands. A positive and strong correlation was noted between progesterone concentration and PI in epithelial cells. The AI remained consistently low throughout the oestrous cycle, with few differences observed across histological components. Caspase-3 labelling displayed the highest positivity in caudal mammary pairs. A negative and moderate correlation was identified between progesterone concentration and AI in interlobular mesenchymal cells. This study highlights the influence of endocrine regulation on cell proliferation indices in mammary tissue, emphasizing the need to consider these hormonal variations in toxicopathological studies involving canine mammary gland.

Pink adipose tissue: A paradigm of adipose tissue plasticity.

Adipose tissue is highly plastic, as illustrated mainly by the transdifferentiation of white adipocytes into beige adipocytes, depending on environmental conditions. However, during gestation and lactation in rodent, there is an amazing phenomenon of transformation of subcutaneous adipose tissue into mammary glandular tissue, known as pink adipose tissue, capable of synthesizing and secreting milk. Recent work using transgenic lineage-tracing experiments, mainly carried out in Saverio Cinti's team, has demonstrated very convincingly that this process does indeed correspond to a transdifferentiation of white adipocytes into mammary alveolar cells (pink adipocytes) during gestation and lactation. This phenomenon is reversible, since during the post-lactation phase, pink adipocytes revert to the white adipocyte phenotype. The molecular mechanisms underlying this reversible transdifferentiation remain poorly understood.

Post-milking application of a Lacticaseibacillus paracasei strain impacts bovine teat microbiota while preserving the mammary gland physiology and immunity.

Bovine mastitis (BM) is a major disease in dairy industry. The current approaches - mainly antibiotic treatments - are not entirely effective and may contribute to antimicrobial resistance dissemination, rising the need for alternative treatment. The present study aims to evaluate the impact of post-milking application of Lacticaseibacillus paracasei CIRM BIA 1542 (Lp1542) on the teat skin (TS) of 20 Holstein cows in mid lactation, in order to reinforce the barrier effect of the microbiota naturally present on the teat. Treatment (Lp1542, iodine or no treatment) was applied post-milking twice a day on the 4 teats of healthy animals for 15 days. Blood and milk samples, and TS swabs were collected at day (D)1, D8, D15 and D26 before morning milking and at D15 before evening milking (D15E) to evaluate Lp1542 impact at the microbial, immune and physiological levels. Lp1542 treatment resulted in a higher lactic acid bacteria and total microbial populations on TS and in foremilk (FM) at D15(E) compared with iodine treatment. Metabarcoding analysis revealed changes in the composition of TS and FM microbiota, beyond a higher Lacticaseibacillus abundance. This included a higher abundance of Actinobacteriota, including Bifidobacterium, and a lower abundance of Pseudomonadota on TS of Lp1542 compared with iodine-treated quarters. In addition, Lp1542 treatment did not trigger any major inflammatory response in the mammary gland, except interleukin 8 production and expression which tended to be slightly higher in Lp1542-treated cows compared with the others. Finally, Lp1542 treatment had no impact on the mammary epithelium functionality (milk yield and composition) and integrity (epithelial cell exfoliation into milk and milk Na+/K+ ratio). Altogether, these results indicate that a topical treatment with Lp1542 is safe with regard to mammary gland physiology and immune system, while impacting its microbiota, inviting us to further explore its effectiveness for mastitis prevention.

Correlation of udder thermogram and somatic cell counts as a tool for detection of subclinical mastitis in buffaloes.

This study aimed to monitor the mammary health of 37 multiparous Murrah buffaloes through infrared thermography (IRT). Based on the California Mastitis Test (CMT) and milk somatic cell counts (SCC), buffaloes were grouped into healthy (H, n = 16), subclinical mastitis (SCM, n = 10), and clinical mastitis (CM, n = 11). Buffaloes were milked twice daily in the morning (5:00-6:00 AM) and evening (5:00-6:00 PM). Rectal temperature and respiratory rates were recorded, CMT was performed and thermal images of the mammary gland of all the buffaloes were taken before and after each milking. Milk samples were analysed after each milking for SCC, fat, Solids-Not-Fat (SNF), density, protein, lactose, salts, conductivity, and pH immediately in the laboratory from fresh milk samples. The surface temperature of the periocular region of both the eyes, muzzle, flank, and vagina were also taken. Thermal images were used to assess the surface temperature of the udder (USST), teat apex (TAT), teat barrel (TB1T), teat base (TB2T), and teat skin surface (TSST). Eye and USST showed significantly higher temperatures (p < 0.05), whereas skin surface temperatures (SST) of different body parts were non-significant in both SCM and CM animals than buffaloes in the H group. Milk SCC showed a positive correlation with conductivity (r > 0.7), salts, and pH (r < 0.6) and a negative correlation with fat, SNF, density, protein, and lactose. TAT, TB1T, TB2T, TSST, and USST were positively correlated with milk SCC. Receiver Operating Characteristic (ROC) analysis of H and SCM groups showed that USST before milking had optimum sensitivity (Se = 0.80) and specificity (Sp = 0.906) among the various skin temperatures recorded. Thermal images captured during the morning showed higher sensitivity compared to images taken in the evening. Results indicate IRT can be used to monitor the mammary health of buffaloes but using IRT in conjunction with milk SCC can help in the accurate prediction of SCM in dairy buffaloes.

The association between teat shape and bimodal milk ejection in Holstein dairy cows.

Our objectives were to investigate the association of teat shape with (1) bimodality, (2) incremental milk flow rates (average flow rates during the first 15 s, 15-30 s, 30-60 s, and 60-120 s of milking), and (3) peak milk flow rate in Holstein dairy cows milked 3 times/d. In this prospective cohort study, we analyzed 220,928 milking observations that were collected from 2,520 cows from a single dairy herd over a period of 31 d. Teat shape was visually assessed and classified into 1 of 4 categories as follows: (1) triangular barrel and pointed teat end (TP), (2) square barrel and round teat end (SR), (3) square barrel, round teat end, and flat in the area of the teat orifice (SRF), and (4) square barrel and flat teat end (SF). Individual cow-level milk flow rates were obtained using electronic on-farm milk meters. We considered bimodality to be present if any of the incremental milk flow rates (flow rates during 15-30 s, 30-60 s, or 60-120 s) were lower than any of the previous rates (flow rates during the first 15 s, 15-30 s, or 30-60 s). The frequency distribution of cows with different teat shapes was as follows: TP, 96 (3.8%); SR, 1,751 (69.5%); SRF, 617 (24.5%); and SF, 56 (2.2%). A generalized linear mixed model revealed differences in the odds of bimodality among cows with different teat shapes. Compared with cows in category SR, the odds (95% CI) of bimodality of cows in other categories were as follows: TP, 0.68 (0.48-0.97); SF, 1.96 (1.21-3.19); and SRF, 1.46 (1.23-1.72). General linear mixed models indicated an association between teat shape and all incremental milk flow rates, with higher milk flow rates being present for cows with teat shapes in the SRF and SF categories. The general linear mixed model for the dependent variable peak milk flow rate indicated an association between teat shape and peak milk flow rate. Least squares means and 95% CI were 5.1 (4.9-5.3) kg/min for TP, 5.8 (5.5-6.1) kg/min for SF, 5.6 (5.5-5.7) for SRF, and 5.3 (5.3-5.4) for SR, respectively. We conclude that in this study cohort, bimodality is more likely to occur in cows with flat teat ends compared with those with a round teat ends. We attribute this relationship to the difference in milking speed across cows with different teat shapes. Teat shape may serve as a useful phenotype to identify cows that are more likely to exhibit bimodality. Because our study population was unique, future studies considering different circumstances such as breed, milking system, and milking routine are needed before results can be extrapolated.

Going Full TeRM: The Seminal Role of Tissue-Resident Macrophages in Organ Remodeling during Pregnancy and Lactation.

During pregnancy and lactation, the uterus and mammary glands undergo remarkable structural changes to perform their critical reproductive functions before reverting to their original dormant state upon childbirth and weaning, respectively. Underlying this incredible plasticity are complex remodeling processes that rely on coordinated decisions at both the cellular and tissue-subunit levels. With their exceptional versatility, tissue-resident macrophages play a variety of supporting roles in these organs during each stage of development, ranging from maintaining immune homeostasis to facilitating tissue remodeling, although much remains to be discovered about the identity and regulation of individual macrophage subsets. In this study, we review the increasingly appreciated contributions of these immune cells to the reproductive process and speculate on future lines of inquiry. Deepening our understanding of their interactions with the parenchymal or stromal populations in their respective niches may reveal new strategies to ameliorate complications in pregnancy and breastfeeding, thereby improving maternal health and well-being.

Conductivity of mammary gland secretions is a sensitive and specific predictor of parturition in mares.

BACKGROUND: Accurate prediction of parturition is paramount to ensuring monitoring of delivery and preventing complications. Assessing the pH and electrolytes of the mammary gland secretions (MGS) helps detect impending parturition. As conductivity is related to electrolyte concentrations and pH, it could be a useful alternative for predicting impending parturition; however, this hypothesis warrants a critical assessment. OBJECTIVES: To assess the ability of conductivity, pH, and Brix in the MGS to predict parturition and to investigate their associations. STUDY DESIGN: Field study. METHODS: The MGS of periparturient mares (n = 241) was assessed daily for conductivity, pH, and Brix index from 320d until parturition. Receiving operating curve cut-off values for conductivity (≤4.8 mS/cm), pH (≤6.4), and Brix index (>23.6%) were used to calculate sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for predicting parturition in ≤24 h. RESULTS: Impending parturition was associated with a reduction in conductivity and pH (p < 0.05), and conductivity was strongly correlated with pH (r = 0.88) and Brix (r = -0.80) (p < 0.05). Sensitivity, specificity, PPV, and NPV for parturition in ≤24 h for conductivity (82%, 91%, 77%, and 92%, respectively), pH (79%, 84%, 81%, and 71%, respectively), and Brix (72%, 79%, 66%, and 83%, respectively) were determined separated and pairwise. Of interest, the sensitivity, specificity, PPV, and NPV, of combining conductivity and pH, were 80%, 95%, 90%, and 88%, respectively. Conductivity (≤4.8 mS/cm) presented the greatest odds ratio for predicting parturition in ≤24 h, and coupling it with pH (≤6.4 pH units) doubled its odds ratio (i.e., 25.4-62.3). MAIN LIMITATIONS: Field study. CONCLUSION: The conductivity of MGS is a sensitive and specific method to predict parturition. This is the first large-scale study showing that a combination of conductivity and pH is useful for predicting parturition in mares. The methods employed can likely apply to other settings with similar results.

Understanding mammary activity in red-rumped agouti and implications for management and conservation of this Neotropical game species / Compreendendo a atividade mamária na cutia e as implicações para o manejo e a conservação desta espécie cinegética Neotropical

Abstract The red-rumped agouti (Dasyprocta leporina) produces precocial young and is the most hunted and farmed game species in several Neotropical countries. An understanding of the reproductive biology, including the relationship between litter size and teat functionality is crucial for conservation management of this animal. In precocial mammals, as the red-rumped agouti, maintaining maternal contact to learn foraging patterns may be more important than the energy demands and nutritional constraints during lactation and suckling may not play important roles when compared to altricial mammals. Therefore, in this study we evaluated the relationship between mammary functionality with litter size, litter birth weight, and parturition number in captive red-rumped agouti. Functionality was assessed by manual palpation of teats from un-sedated females (N=43). We compared the average birth weight of all newborns, male newborns and female newborns among agoutis with different litter sizes and different parturitions by one way ANOVA's, while Pearson's Chi-squared tests were used to detect relationships between teat functionality, litter size, and parturition number. Parturition number had no effect on the mean birth weight of all young (F0.822, P > 0.05), male young (F0.80, P > 0.05) or female young (F0.66, P > 0.05) in the litters. We found (i) no significant correlations (P > 0.05) between teat functionality and litter size and (ii) no significant correlations (P > 0.05) between teat functionality and parturition number. This suggests that whilst all teat pairs were functional, functionality was a poor indicator of litter size; suggesting that female agouti young may not have a high dependency on maternal nutrition; an possible evolutionary strategy resulting in large wild populations; hence its popularity as a game species.

Resumo A cutia (Dasyprocta leporina) produz filhotes precoces, e é a espécie cinegética mais caçada e criada em muitos países neotropicais. O entendimento da biologia reprodutiva, incluindo a relação entre o tamanho da ninhada e a funcionalidade das glândulas mamárias é crucial para o manejo conservacionista deste animal. Nos mamíferos precoces, como a cutia, manter o contato materno para a aprendizagem de padrões de forrageamento pode ser mais importante do que a demanda energética e de restrição nutricional durante a lactação, e a amamentação pode não desempenhar um papel importante quando comparado aos mamíferos altriciais. Portanto, neste estudo nós avaliamos a relação entre a funcionalidade mamária com o tamanho da ninhada, peso ao nascer da ninhada, e o número de partos em cutias cativas. A funcionalidade foi avaliada por meio de palpação manual das glândulas mamárias em fêmeas não sedadas (N=43). Nós comparamos a média do peso ao nascer de todos os recém-nascidos, macho e fêmeas recém-nascidos entre cutias com diferentes tamanhos de ninhadas e diferentes parturições, por meio da análise one-way ANOVA unifatorial; enquanto o teste do chi-quadrado de Pearson foi utilizado para detectar as relações entre a funcionalidade da glândula mamária, o tamanho da ninhada e o número de partos. O número de partos teve efeito na média de peso ao nascer dos filhotes (F0,822, P > 0,05), filhotes machos (F0,80, P > 0,05) ou filhotes fêmeas (F0,66, P > 0,05) nas ninhadas. A análise bivariada de Pearson (i) não mostrou correlação (P > 0,05) entre a funcionalidade da glândula mamária e o tamanho da ninhada e (ii) nenhuma correlação significativa (P > 0,05) entre a funcionalidade da glândula mamária e o número de parições. Os resultados sugerem que, embora todos os pares de tetas fossem funcionais, as cutias recém-nascidas podem não depender da nutrição materna para sobreviver; .embora todos os pares de tetas sejam funcionais, as cutias recém nascidas aparentemente não dependem da nutrição materna para sua sobrevivência; esta é, provavelmente, uma estratégia evolutiva que resulta em grandes populações da cutia em vida livre; e que pode explicar sua popularidade como espécie cinegética

Establishment of a standard operating procedure for predicting the time of calving in cattle

Precise calving monitoring is essential for minimizing the effects of dystocia in cows and calves. We conducted two studies in healthy cows that compared seven clinical signs (broad pelvic ligaments relaxation, vaginal secretion, udder hyperplasia, udder edema, teat filling, tail relaxation, and vulva edema) alone and in combination in order to predict the time of parturition. The relaxation of the broad pelvic ligaments combined with teat filling gave the best values for predicting either calving or no calving within 12 h. For the proposed parturition score (PS), a threshold of 4 PS points was identified below which calving within the next 12 h could be ruled out with a probability of 99.3% in cows (95.5% in heifers). Above this threshold, intermitted calving monitoring every 3 h and a progesterone rapid blood test (PRBT) would be recommended. By combining the PS and PRBT (if PS > or = 4), the prediction of calving within the next 12 h improved from 14.9% to 53.1%, and the probability of ruling out calving was 96.8%. The PRBT was compared to the results of an enzyme immunoassay (sensitivity, 90.2%; specificity, 74.9%). The standard operating procedure developed in this study that combines the PS and PRBT will enable veterinarians to rule out or predict calving within a 12 h period in cows with high accuracy under field conditions.