Hypothalamic Neuromodulation and Control of the Dermal Surface Temperature of Livestock during Hyperthermia.

Hyperthermia elicits several physiological and behavioral responses in livestock to restore thermal neutrality. Among these responses, vasodilation and sweating help to reduce core body temperature by increasing heat dissipation by radiation and evaporation. Thermoregulatory behaviors such as increasing standing time, reducing feed intake, shade-seeking, and limiting locomotor activity also increase heat loss. These mechanisms are elicited by the connection between peripheral thermoreceptors and cerebral centers, such as the preoptic area of the hypothalamus. Considering the importance of this thermoregulatory pathway, this review aims to discuss the hypothalamic control of hyperthermia in livestock, including the main physiological and behavioral changes that animals adopt to maintain their thermal stability.

Use of Infrared Thermography and Heart Rate Variability to Evaluate Autonomic Activity in Domestic Animals.

Most of the responses present in animals when exposed to stressors are mediated by the autonomic nervous system. The sympathetic nervous system, known as the one responsible for the "fight or flight" reaction, triggers cardiovascular changes such as tachycardia or vasomotor alterations to restore homeostasis. Increase in body temperature in stressed animals also activates peripheral compensatory mechanisms such as cutaneous vasodilation to increase heat exchange. Since changes in skin blood flow influence the amount of heat dissipation, infrared thermography is suggested as a tool that can detect said changes. The present review aims to analyze the application of infrared thermography as a method to assess stress-related autonomic activity, and their association with the cardiovascular and heart rate variability in domestic animals.

Hypothalamic Neuromodulation of Hypothermia in Domestic Animals.

When an organism detects decreases in their core body temperature, the hypothalamus, the main thermoregulatory center, triggers compensatory responses. These responses include vasomotor changes to prevent heat loss and physiological mechanisms (e.g., shivering and non-shivering thermogenesis) for heat production. Both types of changes require the participation of peripheral thermoreceptors, afferent signaling to the spinal cord and hypothalamus, and efferent pathways to motor and/or sympathetic neurons. The present review aims to analyze the scientific evidence of the hypothalamic control of hypothermia and the central and peripheral changes that are triggered in domestic animals.

Strategies and Mechanisms of Thermal Compensation in Newborn Water Buffaloes.

Hypothermia is one of the principal causes of perinatal mortality in water buffaloes and can range from 3% to 17.9%. In ruminants, factors affecting hypothermia in newborns may be of intrinsic (e.g., level of neurodevelopment, birth weight, vitality score, amount of brown fat, skin features) or extrinsic origin (e.g., maternal care, environmental conditions, colostrum consumption). When newborn buffaloes are exposed to cold stress, thermoregulatory mechanisms such as peripheral vasoconstriction and shivering and non-shivering thermogenesis are activated to prevent hypothermia. Due to the properties of infrared thermography (IRT), as a technique that detects vasomotor changes triggered by a reduction in body temperature, evaluating the central and peripheral regions in newborn buffaloes is possible. This review aims to analyze behavioral, physiological, and morphological strategies and colostrum consumption as thermal compensation mechanisms in newborn water buffalo to cope with environmental changes affecting thermoneutrality. In addition, the importance of monitoring by IRT to identify hypothermia states will be highlighted. Going deeper into these topics related to the water buffalo is essential because, in recent years, this species has become more popular and is being bred in more geographic areas.

Effect of heat stress on TNF-α, TNFRI and TNFRII expression in BLV infected dairy cattle.

High temperatures for extended periods, which do not allow animals to recover from heat stress, affect in particular those BLV-infected animals that carry a high proviral load. For this study, animals were discriminated between BLV (+) and BLV (-), and those belonging to the first group, were classified based on their proviral load. The expression of the inflammatory cytokine TNF-α and its receptors, which play an important role in disease progression, were quantified by qPCR in two different seasons. During the summer, average temperature was 19.8 °C, maximums higher than 30 °C were frequent. Instead, during the autumn, the average temperature was 12.63 °C, and temperatures never exceeded 27 °C. During this season, almost no periods of temperatures exceeded the comfort limit. Our results revealed that the expression levels of TNF-α and its receptors were downregulated in animals with high proviral load. This fact could affect their antiviral response and predispose to viral dissemination; over time, animals with a poorer immune system are prone to acquiring opportunistic diseases. Conversely, animals with LPL maintained their expression profile, with behavior comparable to non-infected animals. These findings should be considered by producers and researchers, given the problems that global warming is causing lately to the planet.

Hepatic hematopoiesis in the alpaca (Vicugna pacos), a species with development in hypoxic environments.

Hematopoiesis occurs in different anatomical niches throughout the life of the individual. The first hematopoietic extra-embryonic stage is replaced by a intra-embryonic stage that occurs in a region that is adjacent to the dorsal aorta. Then, the prenatal hematopoietic function is continued by the liver and spleen, and later by the bone marrow. The objective of the present work was to describe the morphological characteristics of hepatic hematopoiesis in the alpaca and to analyze the proportion of the hematopoietic compartment of the organ and the cell types, at different times of ontogeny. Sixty-two alpaca samples were collected from the municipal slaughterhouse of Huancavelica, Perú. They were processed by routine histological techniques. Hematoxylin-eosin staining, special dyes, immunohistochemical techniques and supplementary analyses by lectinhistochemistry, were performed. The prenatal liver is an important structure in the expansion and differentiation of hematopoietic stem cells. Their hematopoietic activity was characterized by four stages: initiation, expansion, peak, and involution. The liver started its hematopoietic function at 21 days EGA and it was maintained until shortly before birth. Differences were found in the proportion and morphology of the hematopoietic tissue in the different groups corresponding to each gestational stage.

Neurophysiology of Milk Ejection and Prestimulation in Dairy Buffaloes.

The present review aims to integrate the anatomical characteristics of the mammary gland and the neurophysiology of milk ejection to understand the milking capacity of the water buffalo. Since one of the main uses of this species is milk production, this article will analyze the controversies on the use of oxytocin as a stimulant during milking as well as the existing alternatives that farmers apply to promote correct stimulation during milk letdown. According to the available literature, the efficiency of the milking process, the quality of the milk, and the health of the animals are elements that require the consideration of species-specific characteristics to enhance the performance of buffaloes. The incorporation of technological innovations and competitive strategies could contribute to a better understanding of water buffalo in the milk industry.

Cytokine TNF-α and its receptors TNFRI and TNFRII play a key role in the in vitro proliferative response of BLV infected animals.

Bovine leukemia virus (BLV) main host cells are B lymphocytes. Infected animals can be classified into high or low proviral load (HPL or LPL respectively), regarding the number of proviral copies infected lymphocytes they carry. After infection, there is an overexpression of several cytokines, particularly TNF-α, which has a delicate regulation mediated by receptors TNFRI and TNFRII; the first one involved with apoptosis, while the other stimulates cell proliferation. The study aimed to quantify TNF-α and its receptors mRNA expression, and in which extent in vitro proliferation was affected, in peripheral blood mononuclear cells (PBMC) from BLV-infected animals with different proviral loads, after the addition or not of synthetic TNF-α (rTNF-α) for 48 h. PBMC from BLV-infected animals showed spontaneous proliferation after 48 h in culture but did not show changes in proliferation rates after 48 h incubation in the presence of the rTNF-α. TNF-α mRNA expression after 48 h culture without exogenous stimulation was significantly lower, regardless of the proviral load of the donor, compared to non-infected animals. In the LPL animals, the expression of TNF-α mRNA was significantly lower with respect to the control group while the expression of TNFRI mRNA was significantly increased. The HPL animals showed a significant decrease in the expression of TNF-α and TNFRII mRNA respect to the control group. After 48 h incubation with rTNF-α, PBMC from infected animals had different responses: TNF-α and TNFRI mRNA expression was reduced in PBMC from the LPL group compared to the BLV negative group, but no differences were observed in PBMC from the HPL group. TNFRII mRNA expression showed no differences between HPL, LPL, and BLV negative groups, though HPL animals expressed 10.35 times more TNFRI mRNA than LPL. These results support the hypothesis that LPL animals, when faced with viral reactivation, present a pro-apoptotic and anti-proliferative state. However, complementary studies are needed to explain the influence of TNFRII on the development of the HLP profile. On the other hand, exogenous stimulation studies reinforce the hypothesis that BLV infection compromises the immune response of the animals.

Alterations in TNF-α and its receptors expression in cows undergoing heat stress.

Heat stress is one of the environmental factors that most severely affects milk industry, as it has impact on production, immune responses and reproductive performance. The present study was conducted with high-performance Holando-Argentino cows. Our objective was to study TNF-α and its receptors pattern expression in cows from a region characterized by extreme climatic seasonality. Animals were evaluated in three periods: spring (n = 15), summer (n = 14) and autumn (n = 11). Meteorological records from a local station were used to estimate the temperature and humidity index (THI) by means of an equation previously defined. A THI higher than 68 is indicative of stressing conditions. During the summer period, the animals were exposed to 8.5 ±â€¯1.09 h of heat stress, or THI > 68. In spring, stress hours were reduced to 1.4 ±â€¯0.5 every day, while during the autumn, there were no recorded heat stress events. Expression of TNF-α, and its receptors was determined by qPCR. During the summer, TNF-α and its receptors expression diminished drastically compared to the rest of the year, when stressful conditions were infrequent. We conclude that animals that are not physiologically prepared to resist high temperatures might have a less efficient immune response, reinforcing the need to develop new strategies to improve animal welfare.

Lymphocyte proliferation and apoptosis of lymphocyte subpopulations in bovine leukemia virus-infected dairy cows with high and low proviral load.

Bovine leukemia virus (BLV) is one of the most important virus in dairy cattle. The infection behavior follows what we call the iceberg phenomenon: 60% of infected animals do not show clinical signs; 30% develop persistent lymphocytosis (PL); and the remaining 10%, die due to lymphosarcoma. BLV transmission depends on infected cell exchange and thus, proviral load is determinant. Understanding the mechanisms by which cattle governs the control of viral dissemination will be desirable for designing effective therapeutic or preventive strategies for BLV. The development of high proviral load (HPL) or low proviral load (LPL) might be associated to genetic factors and humoral immune responses, however cellular responses are not fully described. It is known that BLV affects cellular homeostasis: proliferation and apoptosis. It is also known that the BLV tropism is directed towards B lymphocytes, and that lymphocytotic animals have elevated amounts of these cells. Usually, when an animal is infected by BLV, the B markers that increase are CD21, CD5 and CD11b. This increase could be related to the modulation of apoptosis in these cells. This is the first work in which animals infected with BLV are classified according to their proviral load and the subpopulations of B and T lymphocytes are evaluated in terms of their percentage in peripheral blood and its stage of apoptosis and viability. PBMCs from HPL animals proliferated more than LPL and non-infected animals. CD11b+/CD5+ lymphocytes in LPL animals presented greater early and late apoptosis than HPL animals and cells of HPL animals had increased viability than LPL animals. Our results confirm that BLV alters the mechanism of apoptosis and proliferation of infected cells.

Stable infection of a bovine mammary epithelial cell line (MAC-T) with bovine leukemia virus (BLV).

Bovine leukemia virus (BLV) is a retrovirus that affects cattle causing a lymphoproliferative disease. BLV infection has been associated with misbalance of the immune response causing a higher incidence of other infections. Mastitis is one of the most important conditions that affect milk production in cattle. The aim of this study was to stably infect a bovine mammary epithelial cell line (MAC-T). MAC-T cell line was successfully infected with BLV and the infection was confirmed by nested PCR, qPCR, immunocytochemistry, western blot and transmission electron microscopy. This is the first report of a bovine mammary epithelial cell line stably infected with BLV. This new cell line could be used as an in vitro model to study the effect of BLV on the immune response in the mammary gland and the relationship with other agents causing mastitis.

Can Bovine Leukemia Virus Be Related to Human Breast Cancer? A Review of the Evidence.

The incidence of breast cancer is continuously increasing worldwide, as influenced by many factors that act synergistically. In the last decade there was an increasing interest in the possible viral etiology of human breast cancer. Since then, many viruses have been associated with this disease (murine mammary tumor virus, MMTV; Epstein-Barr virus, EBV; and human papillomavirus, HPV). Recently, BLV has been identified in human breast cancers giving rise to the hypothesis that it could be one of the causative agents of this condition. BLV is a retrovirus distributed worldwide that affects cattle, causing lymphosarcoma in a small proportion of infected animals. Because of its similarity with human retroviruses like HTLV and HIV, BLV was assumed to also be involved in tumor emergence. Based on this assumption, studies were focused on the possible role of BLV in human breast cancer development. We present a compilation of the current knowledge on the subject and some prospective analysis that is required to fully end this controversy.

A novel association of BoLA DRB3 alleles in BLV infected cattle with different proviral loads / Uma nova associação de alelos de BoLA DRB3 em bovinos infectados com BLV com diferentes cargas provirais

Bovine leukemia virus (BLV) is associated with the most common neoplastic disease of cattle. BLV has a silent dissemination in the herd due to infected cell exchange, thus the concentration of BLV-infected cells in blood should play a major role in the success of viral transmission. Genes from Bovine leukocyte antigen (BoLA), the MHC system of cattle, are associated with genetic resistance and susceptibility to a wide range of diseases, and also with production traits. Some BoLA DRB3.2 allele polymorphisms in Holstein cattle have been associated with resistance or susceptibility to BLV-disease development, or with proviral load (PVL). This investigation studied 107 BLV-infected Argentinean Holstein dairy cows, all of them belonging to one herd. PVL was analysed by qPCR and animals were classified as high proviral load (HPVL, N = 88) and low proviral load (LPVL, N = 19), and BoLA DRB3.2 alleles were genotyped. Alleles BoLA DRB3.2*1501 and *1201 were significantly associated with HPVL (p = 0.0230 and p = 0.0111 respectively), while allele BoLA DRB3.2*0201 was significantly associated with LPVL (p = 0.0030). The present study aims at contributing to the knowledge of the association between BoLA polymorphism and development of a BLV infection profile. Genes that best explain the PVL in this population resulted BoLA DRB3.2*0201 (as a protection factor) and *1501 (as a risk factor). Allelic differences may play an important role in the development of effective immune responses. A better understanding of how BoLA polymorphism contributes to these responses and the establishment of a BLV status is desirable to schedule and evaluate control measures.(AU)

O vírus da leucemia bovina (BLV) está associado à doença neoplásica mais comum do gado bovino. O BLV tem uma disseminação silenciosa no rebanho devido à troca de células infectadas, assim, a concentração de células BLV infectadas no sangue deve desempenhar um papel importante no sucesso da transmissão viral. Os genes do antígeno leucocitário bovino (BoLA), sistema MHC do gado bovino, estão associados à resistência genética e à susceptibilidade a uma ampla gama de doenças, bem como às características da produção. Alguns polimorfismos de alelos de BoLA DRB3.2 em bovinos Holstein têm sido associados à resistência ou susceptibilidade ao desenvolvimento da doença BLV, ou com carga proviral (PVL). Esta investigação avaliou 107 vacas leiteiras da raça Holstein argentina infectadas com BLV e pertencentes a um único rebanho. A PVL foi analisada por qPCR, os animais foram classificados em alta carga proviral (HPVL, N = 88) e baixa carga proviral (LPVL, N = 19), e os alelos BoLA DRB3.2 foram genotipados. Os alelos BoLA DRB3.2*1501 e *1201 estavam significativamente relacionados à HPVL (p = 0,0230 e p = 0,0111, respectivamente), enquanto o alelo BoLA DRB3.2*0201, à LPVL (p = 0,0030). O objetivo deste estudo é contribuir para o conhecimento da associação entre o polimorfismo de BoLA e o desenvolvimento de infecção por BLV. Os genes que melhor explicam a PVL na população analisada resultaram em BoLA DRB3.2*0201 (como fator de proteção) e *1501 (como fator de risco). As diferenças alélicas podem desempenhar um papel importante no desenvolvimento de respostas imunitárias eficazes. Uma melhor compreensão de como o polimorfismo BoLA contribui para estas respostas e o estabelecimento de um estado BLV é desejável para agendar e avaliar as medidas de controle.(AU)

Toll-like receptors, IFN-γ and IL-12 expression in bovine leukemia virus-infected animals with low or high proviral load.

Bovine leukemia virus (BLV) infection is widespread mainly in dairy cattle and 5-10% of infected animals will die due to lymphosarcoma; most cattle remain asymptomatic but 30% develop persistent lymphocytosis (PL). BLV transmission depends on infected cell exchange and thus, proviral load is determinant. Understanding the mechanisms which govern the control of viral dissemination will be desirable for the design of effective therapeutic or preventive strategies for BLV. The development of high proviral load (HPL) or low proviral load (LPL) might be associated to genetic factors and humoral immune responses, however cellular responses are not fully described. We aimed to characterize cytokines and toll-like receptors (TLR) expression related to the proviral load profiles. IFN-γ and IL-12 mRNA expression level was significantly higher in PBMC from infected cattle (LPL n=6 and HPL n=7) compared to uninfected animals (n=5). While no significant differences were observed in IL-12 expression between LPL and HPL group, IFN-γ expression was significantly higher in LPL animals. Infected cattle exhibited higher expression levels of TLR3, 7-9. Animals with HPL had significantly higher expression of TLR7/8 than uninfected cattle. TLR8 and TLR9 were up-regulated in HPL group, and TLR3 was up-regulated in LPL group. This is the first report related to TLR gene expression in BLV infected cattle and represents evidence of the involvement of these receptors in BLV recognition. Further studies on different subpopulations of immune cells may help clarify their role in response to BLV and its consequences on viral dissemination.

Association of TNF-α gene promoter region polymorphisms in bovine leukemia virus (BLV)-infected cattle with different proviral loads.

Tumor necrosis factor alpha (TNF-α) is a pleiotropic cytokine involved in the immune response against viral and other infections. Its expression levels are affected by a polymorphism in the promoter region of the gene. Bovine leukemia virus is a retrovirus that infects cattle and develops two different infection profiles in the host. One profile is characterized by a high number of proviral copies integrated into the host genome and a strong immune response against the virus, while the most relevant property of the other profile is that the number of copies integrated into the host genome is almost undetectable and the immune response is very weak. We selected a population of cattle sufficiently large for statistical analysis and classified them according to whether they had a high or low proviral load (HPL or LPL). Polymorphisms in the promoter region were identified by PCR-RFLP. The results indicated that, in the HPL group, the three possible genotypes were normally distributed and that, in the LPL group, there was a significant association between the proviral load and a low frequency of the G/G genotype at position -824.