Effects on goal directed behavior and habit in two animal models of Parkinson's disease.

Instrumental conditioning involves two different processes: Goal-directed behavior, characterized by its dependence on the causal relationship between action and outcome and the sensitivity of actions to changes in the value of the outcome; and habits, characterized for its persistence and insensitivity to changes after conditioning. It is known that the dopaminergic system is involved in both kind of learning. The present experiments analyzed two animal models of Parkinson's disease. The 6-OHDA model causes selective damage of the catecholaminergic neurons, specifically affecting the dopaminergic neurons in nigro-striatal system. This model simulates degenerative process symptomatology of Parkinson's disease. On the other hand, the LPS model generates an inflammation process in the infusion area. This model simulates the early symptoms of this disorder, including neuroinflammation and microglia activation. In order to validate both parkinsonian models, we studied if 6-OHDA and LPS models cause the same behavioral effects. The results showed that the 6-OHDA model interfered with the process involved in habit formation. In contrast, animals treated with LPS showed a goal-directed learning deficit. Differences between these models could be due to the different effects on Substantia Nigra neurons. 6-OHDA model might disrupt the nigrostriatal pathway, while LPS could interfere on efferences and afferences to Substantia Nigra.

Adipose-derived stem cells decreased microglia activation and protected dopaminergic loss in rat lipopolysaccharide model.

Adult stem cell therapy is being used extensively to rejuvenate damaged tissue. One important tissue source to obtain these cells is adipose, which contains cells called adipose-derived stem cells (ADSCs). These cells have a great therapeutic potential not only for their multipotent properties as well as for immunomodulatory effects on the immune system. Parkinson's disease is characterized as neurodegenerative disorder which etiology is undoubtedly related to neuroinflammation process. The properties of ADSCs can be used as a new tool in stem cells therapy to treat neurodegenerative disorders. However, their efficacies are still controversial. Some authors have reported neuroprotection effects, while others did not find differences or stem cells increased the damage. Our previous study showed that ADSCs can survive long time after transplantation, suggesting us some biological effects could need more time to be repaired. In this study, we assessed the neuroprotection 6 months after transplantation. Our results suggest ADSCs can protect the dopaminergic loss after lipopolysaccharide (LPS) injection both reducing the microglia activation and differentiating into dopaminergic cells.

Dysregulation of the Hippo pathway signaling in aging and cancer.

Human beings are facing emerging degenerative and cancer diseases, in large part, as a consequence of increased life expectancy. In the near future, researchers will have to put even more effort into fighting these new challenges, one of which will be prevention of cancer while continuing to improve the aging process through this increased life expectancy. In the last few decades, relevance of the Hippo pathway on cancer has become an important study since it is a major regulator of organ size control and proliferation. However, its deregulation can induce tumors throughout the body by regulating cell proliferation, disrupting cell polarity, releasing YAP and TAZ from the Scribble complexes and facilitating survival gene expression via activation of TEAD transcription factors. This pathway is also involved in some of the most important mechanisms that control the aging processes, such as the AMP-activated protein kinase and sirtuin pathways, along with autophagy and oxidative stress response/antioxidant defense. This could be the link between two tightly connected processes that could open a broader range of targeted molecular therapies to fight aging and cancer. Therefore, available knowledge of the processes involved in the Hippo pathway during aging and cancer must necessarily be well understood.

Cell tracking, survival, and differentiation capacity of adipose-derived stem cells after engraftment in rat tissue.

Adipose tissue is an important source of adipose derived stem cells (ADSCs). These cells have the potential of being used for certain therapies, in which the main objective is to recover the function of a tissue/organ affected by a disease. In order to contribute to repair of the tissue, these cells should be able to survive and carry out their functions in unfavorable conditions after being transplanted. This process requires a better understanding of the biology involved: such as the time cells remain in the implant site, how long they stay there, and whether or not they differentiate into host tissue cells. This report focuses on these questions. ADSC were injected into three different tissues (substantia nigra, ventricle, liver) and they were tracked in vivo with a dual GFP-Luc reporter system. The results show that ADSCs were able to survive up to 4 months after the engraftment and some of them started showing resident cell tissue phenotype. These results demonstrate their long-term capacity of survival and differentiation when injected in vivo.

Immunolocalization of Substance P and NK-1 Receptor in ADIPOSE Stem Cells.

Substance P (SP) is a neuropeptide belonging to the thachykinin peptide family. SP, after binding to its receptor, the neurokinin 1 receptor (NK1R), controls several transcription factors such as NF-κB, hypoxia inducible factor (HIF-1α), c-myc, c-fos, c-jun, and AP-1. SP and NK1R have a widespread distribution in both the central and peripheral nervous systems. They are also present in cells not belonging to the nervous system (immune cells, placenta, etc.). SP is located in all body fluids, that is, blood, cerebrospinal fluid, etc., making it ubiquitous throughout the human body. SP and NK1R genes are expressed in the stem cell line TF-1 and in primary stem cells derived from human placental cord blood. However, to our knowledge, the presence of SP and the NK1R receptor in adipose stem cells (ADSC) is unknown. We demonstrated by immunofluorescence the localization of SP and NK1R in human and rat ADSC. SP and NK1R are located in both the cytoplasm and the nucleus of these cells. The NK1R is higher in the nucleus than in the cytoplasm of ADSCs. By Western blot we demonstrated the presence of different isoforms of NK1R that have different subcellular locations in the ADSC. SP induces proliferation and mitogenesis through NK1R in ADSCs. These findings reported here for the first time suggest an important role for a SP/NK1R system, either as genetic and/or epigenetic factor, in both the cytoplasm and nucleus functions of the ADSCs. J. Cell. Biochem. 118: 4686-4696, 2017. © 2017 Wiley Periodicals, Inc.

Aging and Oxidative Stress Decrease Pineal Elongation Factor 2: In Vivo Protective Effect of Melatonin in Young Rats Treated With Cumene Hydroperoxide.

We studied the alterations of Elongation Factor 2 (eEF2) in the pineal gland of aged rats as well as the possible protective role of exogenous melatonin on these changes in young rats treated with cumene hydroperoxide (CH), a compound that promotes lipid peroxidation and inhibits protein synthesis. The study was performed using male Wistar rats of 3 (control), 12, and 24 months and 3-month-old rats treated with CH, melatonin, and CH plus melatonin. We found that pineal eEF-2 is affected by aging and CH, these changes being prevented by exogenous melatonin in the case of CH-treated rats. The proteomic studies show that many other proteins are affected by aging and oxidative stress in the pineal gland. The results suggest that one of the possible mechanisms underlying pineal gland dysfunction during aging is the effect of lipid peroxidation on eEF-2, which is a key component of protein synthesis machinery. J. Cell. Biochem. 118: 182-190, 2017. © 2016 Wiley Periodicals, Inc.

Chronic stress enhances microglia activation and exacerbates death of nigral dopaminergic neurons under conditions of inflammation.

BACKGROUND: Parkinson's disease is an irreversible neurodegenerative disease linked to progressive movement disorders and is accompanied by an inflammatory reaction that is believed to contribute to its pathogenesis. Since sensitivity to inflammation is not the same in all brain structures, the aim of this work was to test whether physiological conditions as stress could enhance susceptibility to inflammation in the substantia nigra, where death of dopaminergic neurons takes place in Parkinson's disease. METHODS: To achieve our aim, we induced an inflammatory process in nonstressed and stressed rats (subject to a chronic variate stress) by a single intranigral injection of lipopolysaccharide, a potent proinflammogen. The effect of this treatment was evaluated on inflammatory markers as well as on neuronal and glial populations. RESULTS: Data showed a synergistic effect between inflammation and stress, thus resulting in higher microglial activation and expression of proinflammatory markers. More important, the higher inflammatory response seen in stressed animals was associated with a higher rate of death of dopaminergic neurons in the substantia nigra, the most characteristic feature seen in Parkinson's disease. This effect was dependent on glucocorticoids. CONCLUSIONS: Our data demonstrate that stress sensitises midbrain microglia to further inflammatory stimulus. This suggests that stress may be an important risk factor in the degenerative processes and symptoms of Parkinson's disease.

In vitro and in vivo protection by melatonin against the decline of elongation factor-2 caused by lipid peroxidation: preservation of protein synthesis.

As organisms age, a considerable decrease in protein synthesis takes place in all tissues. Among the possible causes of the decline of translation in old animals are the modifications of elongation factor-2 (eEF-2). eEF-2 occupies an essential role in protein synthesis where it catalyzes the ribosomal translocation reaction. eEF-2 is particularly sensitive to increased oxidative stress. However, all oxidants do not affect eEF-2, only compounds that increase lipid peroxidation. As peroxides are unstable compounds, they decompose and generate a series of highly reactive compounds, including aldehydes malondialdehyde (MDA) and 4-hydroxynoenal (HNE). We have previously reported that hepatic eEF-2 forms adducts with low-molecular weight aldehydes, MDA and HNE. Therefore, the protection of eEF-2 must be specifically carried out by a compound with lipoperoxyl radical-scavenging features such as melatonin. In this article, we show the ability of melatonin to protect against the changes that occur in the eEF-2 under conditions of lipid peroxidation induced by cumene hydroperoxide (CH), a compound used experimentally to induce lipid breakdown. As experimental models, we used cultured cells and rats treated with this oxidant compound. eEF-2 levels, adduct formation of this protein with MDA and HNE, and lipid peroxides were determined. In the cultured cells, protein synthesis rate was also measured. Our results show that melatonin prevented the molecular changes in eEF-2 and the decline in protein synthesis rate secondary to lipid peroxidation. The results also show that serum levels of several hormones were affected by CH-induced oxidative stress, which was partially or totally prevented by melatonin.

The Neurokinin-1 Receptor Is Essential for the Viability of Human Glioma Cells: A Possible Target for Treating Glioblastoma.

Background: Glioblastoma or glioma is the most common malignant brain tumor. Patients have a prognosis of approximately 15 months, despite the current aggressive treatment. Neurokinin-1 receptor (NK-1R) occurs naturally in human glioma, and it is necessary for the tumor development. Objective: The purpose of the study was to increase the knowledge about the involvement of the substance P (SP)/NK-1R system in human glioma. Methods: Cellular localization of NK-1R and SP was studied in GAMG and U-87 MG glioma cell lines by immunofluorescence. The contribution of both SP and NK-1R to the viability of these cells was also assessed after applying the tachykinin 1 receptor (TAC1R) or the tachykinin 1 (TAC1) small interfering RNA gene silencing method, respectively. Results: Both SP and the NK-1R (full-length and truncated isoforms) were localized in the nucleus and cytoplasm of GAMG and U-87 MG glioma cells. The presence of full-length NK-1R isoform was mainly observed in the nucleus, while the level of truncated isoform was higher in the cytoplasm. Cell proliferation was decreased when glioma cells were transfected with TAC1R siRNA, but not with TAC1. U-87 MG cells were more sensitive to the effect of the TAC1R inhibition than GAMG cells. The decrease in the number of glioma cells after silencing of the TAC1R siRNA gene was due to apoptotic and necrotic mechanisms. In human primary fibroblast cultured cells, TAC1R silencing by siRNA did not produce any change in cell viability. Conclusions: Our results show for the first time that the expression of the TAC1R gene (NK-1R) is essential for the viability of GAMG and U-87 MG glioma cells. On the contrary, the TAC1R gene is not essential for the viability of normal cells, confirming that NK-1R could be a promising and specific therapeutic target for the treatment of glioma.

Genetic selection indices for growth traits in Blanco-Orejinegro cattle.

Selection indices are used in genetic improvement programs, with the purpose of selectins simultaneous for several economically important traits. The objective of this study was to construct equations for selection indices in the Blanco-Orejinegro (BON) breed and to determine the index that would generate the greatest genetic progress. The information used included birth weight (BW), body weights adjusted to 120, 240, 480, and 720 days old (W120, W240, W240, 480 and W720, respectively), age at first calving (AFC) and interval between first and second calving (IBC) estimated breeding values. Two Smith and Hazel indices were calculated using variances (I1) and literature (I2), with a part two indices designed using information from experts and breeders (I3 and I4). All the indices favored the reduction of weight at birth. The I1 focused mainly on W120 and I2, I3 and I4 focused on 720. In general, the estimated indices obtained similar reliability and expected genetic differences I1 generated a decrease in direct BW. I2 generated the largest increases in BW and AFC. I3 and I4 generated positive changes in growth and reproductive traits, with I3 generating the greatest genetic gains in the population, especially for W240.

Effect of Age and Lipoperoxidation in Rat and Human Adipose Tissue-Derived Stem Cells.

A wide range of clinical applications in regenerative medicine were opened decades ago with the discovery of adult stem cells. Highly promising adult stem cells are mesenchymal stem/stromal cells derived from adipose tissue (ADSCs), primarily because of their abundance and accessibility. These cells have multipotent properties and have been used extensively to carry out autologous transplants. However, the biology of these cells is not entirely understood. Among other factors, the regeneration capacity of these cells will depend on both their capacity of proliferation/differentiation and the robustness of the biochemical pathways that allow them to survive under adverse conditions like those found in damaged tissues. The transcription factors, such as Nanog and Sox2, have been described as playing an important role in stem cell proliferation and differentiation. Also, the so-called longevity pathways, in which AMPK and SIRT1 proteins play a crucial role, are essential for cell homeostasis under stressful situations. These pathways act by inhibiting the translation through downregulation of elongation factor-2 (eEF2). In order to deepen knowledge of mesenchymal stem cell biology and which factors are determinant in the final therapeutic output, we evaluate in the present study the levels of all of these proteins in the ADSCs from humans and rats and how these levels are affected by aging and the oxidative environment. Due to the effect of aging and oxidative stress, our results suggest that before performing a cell therapy with ADSCs, several aspects reported in this study such as oxidative stress status and proliferation and differentiation capacity should be assessed on these cells. This would allow us to know the robustness of the transplanted cells and to predict the therapeutic result, especially in elder patients, where probably ADSCs do not carry out their biological functions in an optimal way.

Genetic parameters and trends for growth traits in Blanco Orejinegro cattle.

Since 1940, efforts have been made to preserve the Blanco Orejinegro (BON) cattle breed by maintaining gene banks. Nine years ago, a BON genetic improvement program was implemented to increase genetic gain based on control of productivity and the use of performance tests, polygenic and genomic evaluations, and selection indices. The objective of this study was to estimate genetic parameters and trends for growth traits by using polygenic (PM) and genomic-polygenic (GPM) models. Productive information from the years 1980 to 2019 was used. The following data were used: 7,304 birth weight (BW) records, 1,281 records of body weight adjusted to 120 days (W120), 4,791 records of weight adjusted to 240 days (W240), 3,339 records of weight adjusted to 480 days (W480), and 1,364 records of weight adjusted to 720 days (W720). The relationship matrix included 13,612 pure animals belonging to the Colombian Corporation for Agricultural Research - AGROSAVIA and the Producer Network program. A total of 1,224 individuals were genotyped with chips of different densities and all were imputed up to 50,932 SNPs. Polygenic and genomic-polygenic models were used to estimate genetic parameters and correlations between the genetic values estimated by each model. A generalized additive model with smoothing was used to estimate trends of genetic values from 1980 to 2019. Heritabilities between 0.29 and 0.40, genetic correlations between 0.13 and 0.94, and phenotypic correlations between 0.23 and 0.72 were observed. Close-to-zero genetic growth was observed for BW and W120. Genetic trends for the other traits were positive, with higher growth in the last 7 years. The heritabilities observed in this population indicate that the traits measured would respond to selection. Greater genetic progress can be achieved in W240, W480, and W720 by continuing with genetic evaluations and developing performance tests and strategies to make genetic material readily available to producers.

Advantages and disadvantages of apoptosis in the aging process.

Researchers cannot predict as yet how long a human being can live. Life expectancy has been steadily increasing in the last century, but perhaps not always the quality of life in parallel with it. Future generations will be faced with the problems of an increased life expectancy along with the emergence of new age-related diseases. A deeper understanding of the aging process is crucial to ameliorate, if not to prevent, these projected new old-age diseases. One of the mechanisms responsible for healthy aging is through the effective maintenance of physiological, biochemical, and immunological functions. To carry this out, the organism needs to create new cells to replace old ones and to induce the disappearance of old and damaged cells. Apoptosis is involved in all these processes. However, if apoptosis is dysregulated, premature senescence-associated diseases are likely to appear. In our review, the focus will be on a better understanding of the role of apoptosis in the aging process. These signaling pathways will most assuredly be pharmacologically targeted in antiaging medicine therapies.

Population genetic structure of two herds of Aberdeen Angus cattle breed in Colombia / Estructura genética poblacional de dos hatos de ganado bovino Aberdeen Angus en Colombia / Estrutura genética da população de dois rebanhos da raça Aberdeen Angus na Colômbia

Abstract Background: Two biotypes of Aberdeen Angus cattle breed, known as Old Type and New Type, that differ in their origin and beef production are formally recognized. In Colombia, this breed has been commercialized for approximately 80 years. Studies on the origin, kinship and levels of genetic diversity of this breed in Colombian herds are scarce, yet important for planning crossing and management strategies. Objective: To measure the genetic diversity and structure of two Colombian herds of Old Type and New Type biotypes of Aberdeen Angus from Huila and Cundinamarca provinces and assess mitochondrial introgression with other breeds. Methods: A set of ten microsatellites and sequences of the Mitochondrial Control Region were characterized. Estimators of genetic diversity and population differentiation along with tests of population assignment were applied. Results: Nuclear loci were highly polymorphic as shown by the Polymorphic Information Content (0.599) and the Probability of Identity (1.896 10-08). Both populations were highly diverse and clearly differentiated into two groups corresponding to the Old Type and New Type phenotypes. In contrast, mitochondrial data failed to distinguish these two groups and showed extensive admixture. Conclusions: This study optimized a set of ten highly polymorphic nuclear markers that may be used for parentage and population genetic studies of Aberdeen Angus. Genetic differentiation in these loci agreed with phenotypic differences of the Old and New Types. However, mitochondrial data indicated ancestry of multiple European breeds in the origin of Colombian Aberdeen Angus.

Resumen Antecedentes: Dentro de la raza Aberdeen Angus existen dos biotipos conocidos como Old Type y New Type, las cuales difieren en su origen y producción de carne. En Colombia, esta raza se ha venido comercializando desde hace aproximadamente 80 años. No obstante, aún no se han realizado estudios sobre su origen, parentesco y niveles de diversidad genética de esta raza en hatos colombianos, lo cual es importante para planear estrategias de cruce y manejo. Objetivo: Medir la diversidad y estructura genética de dos hatos colombianos de Aberdeen Angus Old Type y New Type de Huila y Cundinamarca y evaluar la introgresión mitocondrial con otras razas. Métodos: Se caracterizó un grupo de diez loci microsatélite y se secuenció la Región Control Mitocondrial. Se aplicaron estimadores de diversidad genética y diferenciación poblacional, junto con pruebas de asignación poblacional. Resultados: Los loci microsatélite fueron altamente polimórficos, tal como lo indicaron el Contenido de Información Polimórfica (0,599) y la Probabilidad de Identidad (1,896 10-08). Las poblaciones evaluadas de Aberdeen Angus en Colombia fueron altamente diversas y se diferenciaron claramente en dos grupos correspondientes a los fenotipos Old Type y New Type. En contraste, los datos mitocondriales no recobraron estos dos grupos y mostraron una amplia mezcla genética. Conclusiones: Este estudio optimizó un grupo de diez marcadores altamente polimórficos que pueden ser usados para estudios de parentesco y genética poblacional de Aberdeen Angus. La diferenciación genética en loci nucleares concordó con las diferencias fenotípicas entre Old y New Types, pero los datos mitocondriales indicaron ancestría de múltiples razas europeas en el origen del Aberdeen Angus colombiano.

Resumo Antecedentes: Dentro da raça Aberdeen Angus há dois biótipos conhecidos como Old Type e New Type, que diferem em sua origem e produção de carne. Na Colômbia, esta raça é comercializada há aproximadamente 80 anos. Entretanto, estudos sobre a origem, o parentesco e os níveis de diversidade genética desta raça em rebanhos colombianos ainda não foram realizados, o que é importante para o planejamento de cruzamentos e estratégias de manejo. Objetivo: Medir a diversidade genética e a estrutura de dois rebanhos colombianos de biótipos de Old Type e New Type de Aberdeen Angus de Huila e Cundinamarca e avaliar a introgressão mitocondrial com outras raças. Métodos: Um grupo de dez loci de microssatélites foi caracterizado e a Região de Controle Mitocondrial foi sequenciada. As estimativas de diversidade genética e diferenciação populacional foram aplicadas, juntamente com testes de designação populacional. Resultados: Os locus microssatélites foram altamente polimórficos, conforme indicado pelo Conteúdo de Infomação Polimórfica (0,599) e Probabilidade de Identidade (1,896 10-08). As populações avaliadas de Aberden Angus na Colômbia eram altamente diversificadas e claramente diferenciadas em dois grupos correspondentes aos fenótipos do Old Type e New Type. Em contraste, os dados mitocondriais não recuperaram esses dois grupos e mostraram um amplo mix genético. Conclusões: Este estudo otimizou um grupo de dez marcadores altamente polimórficos que podem ser usados para estudos genéticos de parentesco e população de Aberdeen Angus. A diferenciação genética nos loci nucleares concordou com as diferenças fenotípicas entre os Old e New Types, mas os dados mitocondriais indicam ancestralidade de várias raças européias na origem do Aberdeen Angus colombiano.

Chronic stress alters the expression levels of longevity-related genes in the rat hippocampus.

The molecular mechanisms underlying the negative effects of psychological stress on cellular stress during aging and neurodegenerative diseases are poorly understood. The main objective of this study was to test the effect of chronic psychological stress, and the consequent increase of circulating glucocorticoids, on several hippocampal genes involved in longevity. Sirtuin-1, p53, thioredoxin-interacting protein, and heat shock protein 70 were studied at the mRNA and protein levels in stressed and non-stressed animals. Stress treatment for 10 days decreased sirtuin-1 and heat shock protein 70 levels, but increased levels of p53, thioredoxin-interacting protein and the NADPH oxidase enzyme. Examination of protein expression following two months of stress treatment indicated that sirtuin-1 remained depressed. In contrast, an increase was observed for thioredoxin-interacting protein, heat shock protein 70, p53 and the NADPH oxidase enzyme. The effect of stress was reversed by mifepristone, a glucocorticoid receptor antagonist. These data suggest that chronic stress could contribute to aging in the hippocampus.

Jacob's disease associated with temporomandibular joint dysfunction: a case report.

Jacob's disease is regarded a rare condition in which a joint formation is established between an enlarged mandibular coronoid process and the inner aspect of the zygomatic body. Chronic temporomandibular joint (TMJ) disk displacement has been proposed as etiological factor of coronoid process enlargement. We present a 23-year-old woman with long-standing TMJ dysfunction and restricted interincisal opening, who developed a progressive zygomatic asymmetry. The patient underwent treatment by intraoral coronoidectomy and homolateral TMJ arthroscopy in the same surgery. The histopathological diagnosis of the coronoid sample was cartilage-capped exostoses with presence of articular fibrous cartilage. Although the low prevalence of this entity, it should be considered as a possible diagnosis in patients with progressive limitation of mouth opening, although a TMJ syndrome may be present as a cause of this entity.

Lipid peroxidation: production, metabolism, and signaling mechanisms of malondialdehyde and 4-hydroxy-2-nonenal.

Lipid peroxidation can be described generally as a process under which oxidants such as free radicals attack lipids containing carbon-carbon double bond(s), especially polyunsaturated fatty acids (PUFAs). Over the last four decades, an extensive body of literature regarding lipid peroxidation has shown its important role in cell biology and human health. Since the early 1970s, the total published research articles on the topic of lipid peroxidation was 98 (1970-1974) and has been increasing at almost 135-fold, by up to 13165 in last 4 years (2010-2013). New discoveries about the involvement in cellular physiology and pathology, as well as the control of lipid peroxidation, continue to emerge every day. Given the enormity of this field, this review focuses on biochemical concepts of lipid peroxidation, production, metabolism, and signaling mechanisms of two main omega-6 fatty acids lipid peroxidation products: malondialdehyde (MDA) and, in particular, 4-hydroxy-2-nonenal (4-HNE), summarizing not only its physiological and protective function as signaling molecule stimulating gene expression and cell survival, but also its cytotoxic role inhibiting gene expression and promoting cell death. Finally, overviews of in vivo mammalian model systems used to study the lipid peroxidation process, and common pathological processes linked to MDA and 4-HNE are shown.

Milk quality: milking personnel associated factors / Calidad de la leche: factores asociados al personal vinculado al ordeño

ABSTRACT Objective. To identify factors associated with high and low Somatic Cell Counts (SCC) levels in bulk tanks of dairy farms in Southeast Brazil. Materials and methods. A total of 68 dairy herds with high and low bulk tank SCC levels were analyzed. Surveys and checklists were applied to the personnel regarding milking routines and equipment. Results. Milkers and management personnel explained up to 40.28% of the variability among herds, while the milker's well-being and stability explained up to 28%. Planning and organization were relevant for SCC, as well as the state of the equipment and the milking routine. According to separate analyzes of employees and owners, employees have greater variability in terms of knowledge on milk production, mastitis, milking routine, and experience. Conclusion. There are qualifying factors in milking systems in southeastern Brazil associated with milking personnel, equipment and milking routine. Understanding these factors will enable the implementation of strategies to produce better quality milk.

RESUMEN Objetivo. Identificar factores asociados a altos y bajos niveles de recuentos de células somáticas (RCS) en tanques de hatos lecheros del Sudeste de Brasil. Materiales y métodos. Se analizaron 68 hatos lecheros con niveles altos y bajos de RCS en tanque. Para identificar factores asociados al personal vinculado al ordeño y relacionarlos con RCS se aplicaron encuestas y listas de chequeo para la rutina y el equipo de ordeño. Resultados. El personal vinculado al ordeño, administración y gestión del productor explicaron hasta el 40.28% de la variabilidad entre rebaños, mientras que el bienestar y la estabilidad del ordeñador explicaron hasta el 28%. La planeación y organización del productor fueron relevantes en el RCS, al igual que el estado del equipo y la rutina de ordeño. Análisis separados de empleado y propietario permitieron concluir que existe mayor variabilidad para los primeros, diferenciándose por conocimientos en la producción de leche y el manejo de la mastitis, la rutina y la experiencia. Conclusión. Existen factores clasificatorios en los sistemas de ordeño del sudeste de Brasil asociados al personal, el equipo y la rutina de ordeño. El entendimiento de estos factores posibilitará la implementación de estrategias que permitan producir leche de mejor calidad.

Association of BoLA-DRB3 and TLR4 alleles with subclinical mastitis in cattle from Colombia / Asociación de los alelos BoLA-DRB3 y TLR4 con mastitis subclínica en vacas de Colombia / Associação de Bola-DRB3 alelos e TLR4 com mastite subclínica em vacas da Colômbia

Background: molecular markers for genetic resistance can be used to control mastitis in dairy cattle. The Major Histocompatibility Complex and the Toll-like receptor 4 (TLR4) are two promising genes that warrant investigation. Objective: to identify associations between genotypes of BoLA-DRB3 locus and T4CRBR2 fragment and subclinical mastitis (SM). Methods: 996 lactating cows from 32 herds comprising Holstein (80%), Holstein x Jersey cross (12.5%), and other crosses (7.5%) were evaluated monthly during two years, diagnosed for SM and genotyped for the second exon of BoLA DRB3 and the TLR4 coreceptor-binding region 2 (T4CRBR2) using a Polymerase chain reaction-restriction fragment length polymorphism technique (PCRRFLP). The association between candidate alleles and subclinical mastitis was measured by logistic regression. Results: the most frequently observed alleles for BoLA-DRB3 were DRB3.2 *8, *22, *24, *16, *10, *23, *gba, *11, *2, *mbb, *jba, *3, and *15, accounting for 58.9% of the population. Frequencies for T4CRBR2 alleles A and B were 0.352 and 0.647, respectively. Based on 57,408 observations during the period, the mean SM prevalence was 16.2% (95% CI 13.0 and 19.4) per udder quarter and 37.6% (95% CI 32.1 and 43.2) per cow. The predominant microorganisms isolated from SM quarters were Streptococcus agalactiae and Coagulase-Negative Staphylococci (CNS). Allele DRB3.2 *23 was associated with SM occurrence and CNS infection. No alleles were associated with Streptococcus agalactiae infection. Allele *mbb was associated with occurrence of CNS infection and alleles *jba and *15 were associated with resistance to CNS infection. No significant relationship between T4CRBR2 and SM was observed. Conclusion: DRB3.2 gen may play an important role in the occurrence of SM and certain alleles may confer resistance to specific pathogens.

Antecedentes: los marcadores moleculares genéticos de resistencia para mastitis bovina son una herramienta para el control de la enfermedad en rebaños lecheros. Los genes del Complejo Mayor de Histocompatibilidad y el Receptor tipo Toll 4 (TLR4) son dos genes candidatos promisorios que justifica investigar. Objetivo: identificar asociaciones entre los genotipos del locus BoLA-DRB3 y del fragmento T4CRBR2 con la ocurrencia de mastitis subclínica. Métodos: 996 vacas lactantes de 32 hatos de las razas Holstein (80%), Holstein x Jersey (12,5%) y otros cruces (7,5%), fueron visitadas mensualmente por dos años, diagnosticadas para mastitis subclínica y genotipificadas para el segundo exón del DRB3 y para la región 2 de unión al correceptor del TLR4 (T4CRBR2) por medio de las técnicas de Reacción en cadena de la polimerasa y de Longitud del polimorfismo del fragmento de restricción (PCR-RFLP). La asociación entre los alelos candidatos y la mastitis subclínica se midió por regresión logística. Resultados: los alelos más frecuentes para el DRB3.2 fueron *8, *22, *24, *16, *10, *23, *gba, *11, *2, *mbb, *jba, *3 y *15, que suman el 58,9% del total en la población. Las frecuencias para los alelos A y B del T4CRBR2 fueron de 0,352 y 0,647, respectivamente. Basados en 57.408 observaciones, la prevalencia de MS a nivel de cuarto fue 16,2% (95% IC 13,0 y 19,4) y a nivel de vaca fue de 37,6% (95% IC 32,1 y 43,2). Los microorganismos más frecuentes fueron Streptococcus agalactiae y Estafilococo Coagulasa Negativo (ECN). El alelo DRB3.2 *23 fue el más asociado con la ocurrencia de MS y con la infección por ECN. No se hallaron alelos asociados a infección con mastitis por Streptococcus agalactiae. Con respecto a la infección por ECN, el *mbb se asoció con la ocurrencia y los alelos *jba y *15 se asociaron con resistencia. No se observó asociación entre T4CRBR2 y MS. Conclusión: el gen DRB3.2 puede jugar un papel importante en la presencia de MS y ciertos alelos pueden conferir resistencia a patógenos específicos.

Antecedentes: o uso de marcadores moleculares de resistência para mastites permite controlar esta doença em rebanhos leiteiros. Os genes Toll Like Receptor 4 (TLR4) e o Complexo Mayor de Histocompatibilidade são dois genes candidatos promissórios que justifica pesquisar. Objetivo: identificar associações entre genótipos do locus BoLA-DRB3 e do fragmento T4CRBR2 com a ocorrência de mastite subclínica. Método: 996 vacas em lactação de 32 rebanhos da raça Holandesa (80%), Holandesa x Jersey (12,5%) e outras cruzas (7,5%) foram visitadas mensalmente por dois anos, diagnosticadas para mastites subclínica e genotipadas para o exon segunda BoLA DRB3 e região 2 da ligação co-receptor TLR4 (T4CRBR2) através das técnicas de Reação em Cadeia da Polimerase e do Polimorfismo de Comprimento do Fragmento de Restrição (PCRRFLP). A associação entre alelos candidatos e mastite subclínica foi realizada por meio de regressão logística. Resultados: os alelos mais frequentes *8, *22, *24, *16, *10, *23, *gba, *11, *2, *mbb, *jba, *3 e *15, com um 58,9% do total da população. As frequências dos alelos A e B do T4CRBR2 foram 0,352 e 0,647, respectivamente. Com base em 57.408 observações, a prevalência da SM em quartos mamários foi de 16,2% ((IC 95% 13,0 e 19,4) e ao nível de vaca foi de 37,6% (IC 95% 32,1 e 43,2). Os microrganismos mais comuns foram: Streptococcus agalactiae e Estafilococos Coagulase-negativo, ECN. O alelo DRB3.2 *23 foi o mais associado com a ocorrência de SM e com a infecção por ECN. Não foram encontrados alelos associados à infecção por Streptococcus agalactiae. Em relação à infecção por ECN, o *mbb esteve associado com ocorrência e os alelos *jba e *15 estiveram associados com resistência. Não existiu associação entre MS e os alelos do T4CRBR2. Conclusão: o gene DRB3.2 bovino pode desempenhar um papel importante na presencia de MS e alguns alelos podem conferir resistência à patógenos específicos.

Variance components and genetic parameters for milk production of Holstein cattle in Antioquia (Colombia) using random regression models / Componentes de varianza y parámetros genéticos para producción de leche en ganado Holstein de Antioquia (Colombia), utilizando modelos de regresión aleatoria / Componentes de variância e parâmetros genéticos para produção de leite em gado Holandês da Antioquia, (Colombia) utilizando modelos de regressão aleatória

Background: genetic parameters of lactation curve in dairy cattle can be analyzed as longitudinal data using random regression models (RRM). Objective: the goal of the present study was to estimate variance components and genetic parameters for milk production in Holstein cattle located in Antioquia province using RRM. Methods: a total of 3,158 monthly controls corresponding to 741 first lactations of Holstein cows were evaluated. The RRM included several Legendre polynomials to estimate the population fixed-curve coefficients and to predict the direct additive genetic and the permanent environment effects. Additionally, heterogeneous residual variances were considered by grouping the days in milk into 5 and 10 classes. Eleven models with first to fourth order polynomials were used to describe the direct additive genetic and the permanent environment effects. The residue was modeled by considering five variance classes. Models were compared using Schwartz Bayesian and Akaike's information criteria. Results: the best model was obtained by fourth order Legendre polynomials to estimate the fixed curve of the population, genetic and permanent environment effects. In addition, 5 kinds of days were used to model the residual variances. The variance for the animal genetic, phenotypic, permanent environment, and residual effects decreased as days increased. Milk production heritability in early lactation was 0.36, increasing until 95 days (0.41), with subsequent decrease, reaching 0.10 at 245 days. The permanent environment variance values decreased to 125 days (0.13) and then increased to 215 days (0.21), to finish at the last stage of lactation with values of 0.05. The genetic and phenotypic correlations between milk yields at different days of lactation decreased as days intervals increased. Conclusion: the findings of this study suggest that in the first 150 days of lactation animals better express their genetic potential and that after 180 days there is greater environmental effect.

Antecedentes: los parámetros genéticos de la curva de lactancia en ganado de leche pueden ser analizados como datos longitudinales usando modelos de regresión aleatoria (RRM). Objetivo: el objetivo de este estudio fue estimar componentes de varianza y parámetros genéticos para la producción de leche en vacas Holstein en el departamento de Antioquia, utilizando RRM. Métodos: se utilizaron 3.158 controles mensuales de 741 primeras lactancias. Se usaron RRM con diferentes grados de polinomios de Legendre para estimar los coeficientes de la curva fija de la población y la predicción de los efectos genético aditivo directo y de ambiente permanente y se consideraron 5 y 10 clases de varianzas residuales heterogéneas. Se emplearon once modelos con polinomios de primer a cuarto orden, para describir los efectos genético aditivo directo y ambiente permanente. Los modelos fueron comparados mediante los criterios de información bayesiano de Schwartz y de Akaike. Resultados: el mejor modelo presentó polinomios de cuarto orden 4, 4 y 4 de la curva fija, del efecto genético aditivo y de ambiente permanente, respectivamente, y con 5 clases de varianzas heterogéneas (444.het5). La varianzas para los efectos genético animal, fenotípico, de ambiente permanente y residual disminuyeron con el aumento de los días. La heredabilidad de la producción de leche al inicio de la lactancia fue de 0,36 y fue aumentando hasta los 95 días (0,41), con posterior disminución, llegando a 0,10 a los 245 días. Para la trayectoria de la proporción de ambiente permanente los valores descendieron hasta los 125 días (con 0,13), luego aumentaron hasta los 215 días (con 0,21), para finalizar en la última etapa de la lactancia con valores de 0,05. Las correlaciones genéticas y fenotípicas entre producciones de leche en los diferentes días de lactancia disminuyeron con el aumento del intervalo de los días. Conclusión: los resultados encontrados en este estudio sugieren que en los primeros 150 días de lactancia los animales expresan mejor su potencial, y que despues de 180 días hay mayor impacto ambiental.

Antecedentes: os parâmetros genéticos da curva de lactação em gado leiteiro podem ser analisados como dados longitudinais usando modelos de regressão aleatória (RRM). Objetivo: o objetivo deste estudo foi estimar os componentes de variância e os parâmetros genéticos para produção de leite de vacas holandesas em Antioquia, utilizando um modelo de regressão aleatória (RRM). Métodos: foram utilizados 3.158 controles mensais de 741 primeiras lactações. Usaram-se RRM com diferentes graus de polinômio ortogonal de Legendre para estimar os coeficientes da curva fixa da população e a predição dos efeitos genéticos aditivos diretos e de ambiente permanente. Consideraram-se 5 e 10 classes de variâncias residuais heterogêneas. Foram empregados 11 modelos com polinômios de primeira ate quarta ordem para descrever os efeitos genéticos aditivos diretos e de ambiente permanente. Os modelos foram comparados mediante os critérios de informação bayesiano de Schwartz e de Akaike. Resultados: o melhor modelo foi o de quarto ordem (4, 4 e 4) da curva fixa, do efeito genético aditivo e de ambiente permanente, respectivamente, e com cinco classes de variâncias heterogéneas (444.het5). A variância para os efeitos genético animal, fenotípico, de ambiente permanente e residual diminuiu com o aumento dos dias. A herdabilidade da produção de leite ao inicio da lactação foi de 0.36 e foi aumentando até os 95 dias (0.41), com posterior diminuição, chegando até 0.10 aos 245 dias. Para a trajetória da proporção de ambiente permanente os valores descenderam até os 125 dias (com 0.13), com posterior aumento até os 215 dias (com 0.21), para finalizar na última etapa da lactação com valores de 0.05. As correlaciones genéticas e fenotípicas entre produções de leite nos diferentes dias de lactação diminuíram com o aumento do intervalo dos dias. Conclusão: os resultados encontrados sugerem que nos primeiros 150 dias da lactação os animais expressaram melhor seu potencial genético.