Heat stress is a prominent issue among dairy cattle, primarily due to their heightened vulnerability to fluctuations in surrounding temperature, which is attributed to their increased metabolic rate that is required to produce milk [60]. When dairy cattle experience heat stress, body temperature and respiration rate can be affected [61], resulting in health consequences [8]. In fact, heat stress significantly damages the thermal homeostasis system of the cows and alters body temperature. In study [62], the research findings shed light on the significant impact of heat stress on the thermal homeostasis system of heifers. Heat stress, in this context, refers to a state in which animals experience difficulties in maintaining their normal body temperature as a result of exposure to intense heat. When the surrounding air temperature surpasses 30 °C, it exerts a considerable strain on the heifers’ ability to regulate their body temperature. This manifests in notable increases in both ocular (eye) and skin surface temperatures, which are indicative of their struggle to dissipate excess heat. Similarly, ref. [63] investigated the applicability of Temperature–Humidity Index (THI) for playing a crucial role in evaluating and quantifying heat stress in crossbred dairy animals, particularly in regions with temperate to hot climates. The THI is a composite index that takes into account both temperature and humidity levels, offering a comprehensive measure of the environmental heat load. When the THI value exceeds 80, it signifies a particularly high level of heat stress. Meanwhile, crossbreed cattle only experienced slight to no stress when the THI was below 74, whereas a THI between 74 and 79 resulted in a moderate level of stress.

In agreement with the previous findings, ref. [64] observed that the increment in the internal animal temperature of Angus and Taurine breed adapted (e.g., Romosinuano) bulls exposed to heat stress was induced by a heat load greater than its dissipation capacity. This result was in line with the finding in [54]. In fact, animal production was negatively impacted significantly earlier than the attainment of the critical tolerance threshold because of the body’s difficulty in maintaining a constant temperature. Ref. [18] investigated the effects of heat stress on body temperature in healthy dairy cows early postpartum in two weather periods with distinctly different THIs (i.e., 59.8 ± 3.8 and 74.1 ± 4.4). When a cow perceives thermal discomfort, it begins to alter behavior and endures physiological adaptations [65]. According to investigations in [66], an increase in core body temperature is strongly correlated to the amount of time the cows spend standing in a 24 h period.

Study [58] showed that as the heat load index increases, cows spend less time lying down; the respiration rates and body temperature increase. Indeed, aside from alterations in the temperature of the body, heat stress can lead to a rise in the respiratory rate in dairy cows [67]. As the THI rises, the cattle begin to pant trying to cool the body [68,69]. Panting is a form of the evaporative cooling process that is closely connected to heat stress [10] and, as a quick and severe reaction to heat stress, this behavior occurs in environments with high THI levels [7].

Furthermore, heavy breathing promotes increased CO₂ exhalation, which contributes to rumen acidosis. In fact, the body should maintain a fixed HCO₃ to CO₂ ratio (i.e., 20:1) to operate as an efficient blood pH buffering mechanism [70]. Due to the reduction in blood CO₂ caused by hyperventilation, the kidney secretes HCO₃ in order to maintain this ratio. This limits the HCO₃ quantity available for buffering and sustaining the rumen pH [70].

Monitoring dairy cattle pulse rates can be a useful method for detecting animals affected by heat stress and at risk of developing more significant health issues. With specific reference to heart rate, in [71], the authors investigated the impact of thermal stress in Holstein bull calves during a warm episode in summer to evaluate the immediate physiological reactions of calves to heat stress. Under heat load circumstances, animals housed in shaded pens have a lower heart rate than non-protected animals. This result was supported in [72] for dairy cows in and out of shade. The pulse rate of the animals in the sunlight was much greater than the pulse rate of cows within a shelter. These findings agree with those reported in [62], where it was confirmed that ambient temperature affects heart rate. When the surrounding temperature was higher than 30 °C, heifers under heat stress had a higher heart rate with a 15% rise in the rate of beats per minute. The daily evaluation of native zebu breeds also revealed greater intensities for physiological indicators including heart rate and respiratory rate throughout the afternoon [73]. Similarly, Murrah buffaloes raised in the Indian area of Punjab showed greater pulse rate, respiration rate, and rectal temperature during hot–humid climate conditions [74].

As far as the rumination time in dairy cows is concerned, it has been demonstrated that heat stress can have a significant impact. Several studies have shown that high temperatures reduce the cattle rumination time [75] which is a vital component of the digestive process [76] and plays an important role in the milk yield process [77]. As an example, the study conducted in [78] identified a negative correlation between the THI and the rumination time in Holstein dairy cows. The amount of time spent ruminating decreases as THI rises. As part of this experimental research, six distinct THI thresholds were employed for the categorize instances of thermal stress: “safe” (THI < 68), “mild discomfort” (68 ≤ THI < 72), “discomfort” (72 ≤ THI < 75), “alert” (75 ≤ THI < 79), “danger” (79 ≤ THI < 84), and “emergency“ (THI ≥ 84). This result was confirmed in [13], in which the authors carried out research on Italian Friesian dairy cows raised in a free stall barn, identifying a THI value of 52 as the threshold at which ruminating time decreases.

Furthermore, in ref. [79], the authors reported a rumination time reduction of about 2 min for every daily maximum THI point beyond the threshold value of 76 in 21 Italian Holstein cows. Likewise, in ref. [69], the authors noticed an inverse relationship between rumination time and THI. They identified a decrease in ruminating time during the night under heat stress circumstances. During colder hours, when there is a decrease in the heat stress effects, animals tend to allocate a greater amount of time to ruminating. Dairy cows under heat stress may minimize ruminating activity during cooler hours of the day to efficiently decrease the metabolic temperature [69].

Elevated temperatures can significantly impact reproduction and fertility of dairy cattle [80]. Cows undergo physiological changes when exposed to severe temperatures, which can result in decreased fertility and reproduction potential. When the body temperature increases, the endocrine system suffers, causing hormones to become unbalanced. The impact of heat stress on dairy cows has detrimental effects on their reproductive cycle [81], such as causing premature corpus luteum death, resulting in an augment of infertility and abortion. Furthermore, high temperatures can increase the risk of fetal death. In fact, in ref. [82], the authors observed that progesterone levels and prolactin concentrations were adversely associated with the estrus cycle. In addition to the impacts of extreme heat stress, lower plasma progesterone levels were also a result of hyperprolactinemia, which may have impeded follicular development before ovulation and limited luteal tissue growth after ovulation, resulting in an infertility. Moreover, in ref. [83], the authors deduced that the prolonged luteal phases, evident in heat stress-affected cattle, resulted from insufficient estradiol secretion by a follicle likely impaired by heat stress.

With specific regard to the ovarian pool of oocytes, it is one of the components of the female reproductive tract most vulnerable to heat [51]. High temperatures pose a significant threat to the ovarian pool of oocytes, affecting gene regulation in oocytes and early embryos [84]. According to research in [51], because of their protracted growth process, ovarian follicles and their encapsulated eggs are susceptible to external heat stress throughout the hot months. As a result, heat-induced changes in the early phases of follicular development might manifest later as impaired oocyte maturation and developmental competence. This mechanism might explain why cow fertility decreases in the fall when the cows are no longer subjected to heat stress, implying a carryover impact on the oocyte.

Furthermore, thermal damage to oocytes causes morphological abnormalities, oxidative stress, nuclear fragmentation and mitochondrial dysfunction [85]. In ref. [86], the authors noted the proportion of oocytes reaching the Blastocystis stage was lower in hot–warm seasons compared to that in cool seasons. Heat stress inhibits the production of the luteinizing hormone, estradiol, and the pre-ovulatory surge, which results in poor follicle development and inactive ovaries in cattle [87]. Although it is susceptible to maternal heat stress, the pre-implantation embryo becomes less sensitive as it develops. Heat stress was observed to reduce the fraction of embryos that progressed to the blastocyst stage in lactating cows on Day 1 following estrus, when embryos are one to two cells [88]. Indeed, even mild maternal hyperthermia caused by ambient thermal stress can impair reproductive activities such as estrus, fertilization, embryonic growth and survival [89].

With respect to the conception rate (CR), it has been noted that lactating dairy cows experience a decline throughout the summer season [90]. In study [30], the CR was highly related to the daily average THI on the day of breeding. CR fell steadily, beginning at the lowest THI threshold of 41 and increasing by one unit for each point increase in the average THI on the day of breeding. These data suggest that heat stress occurs in lactating dairy cows also at low THI values and in moderate climates, as indicated by a reduction in CR. 

Because of physical characteristics such as coat color, breeds respond differently to heat stress [91]. For instance, white coats are more tolerant to heat stress compared to animals with dark coats [91]. This is consistent with [92], in which the authors observed that animals with light pigmentation have substantially greater reflectance values for wavelengths between 300 and 850 nm than those with dark coats. A similar finding was reported in study [93], in which the authors observed that Boran cow body temperature gradients were lower than those of Nguni cows. This was most likely owing to increased skin temperature due to high external conditions resulting from vasocontrolled thermoregulation introducing more blood into the skin tissue [94]. This result was in agreement with work [95], the authors of which noted that lactating black cows have a body temperature increase of approximately 4.8 °C when exposed to direct sunshine, whereas it is about 0.7 °C for white cows. This results from the greater absorption of solar radiation by black coats (89%) as opposed to white coats (66%). Analogously, in [41], it was noticed that red and white cattle showed lower temperatures in white areas during the hot season compared to black and white animals. These findings imply that the red coat genotype absorbs less solar energy and then less heat. Deeply pigmented skin shields the underlying tissues from short-wave UV direct radiation by preventing its penetration [54].

Furthermore, coat length, thickness, and hair density influence the adaptation of animals in tropical climates, where short hair, thin skin, and fewer hair follicles per unit area are directly related to increased adaptability to hot temperatures [54]. Ref. [96] explored the association between Holstein cows’ survival rates and hair coat color in hot areas. The authors observed that cows living in the tropics must have a stronger aptitude for evaporative cooling, which is most likely associated to darker pigmentation, hair coat color and physical characteristics. 

Standing and lying events are adaptation mechanisms that can aid both effective dissipation of body heat and heat load reduction from ground surfaces [97]. High temperatures may lead to prolonged standing behavior inducing a reduction in milk production [10,66]. By standing more frequently, cows may improve their respiratory efficiency and properly use their body surface area for a more efficient management of sensible and insensible heat loss, reducing the amount of heat, which they are able to transfer from a heated surface when they are lying [66]. Ref. [98] highlighted that standing during the first 21 days of a lactation can influence the subsequent lying behavior throughout the rest of the lactation period. Regardless of the heat stress level, ref. [38] claimed that difference in core body temperature is minimal, i.e., 0.07 °C, between standing and lying cows. According to the same study, cows were more likely to be lying during the warmest hours of the day, when their body temperature averaged the highest, than during the colder overnight hours. This result agrees with study [99], the authors of which investigated the lying behavior and activity in healthy grazing cows throughout the period from late gestation to early lactation. Cows exposed to cold and rainy weather spent less time lying down. Similarly, the authors of ref. [69] conducted an experimental campaign monitoring cow activity in order to discover behavioral changes induced by heat stress. The work concluded that, when heat stress is present, the amount of daily time spent resting decreases. Ref. [100] revealed a noticeable reduction in the daily lying time equal to 3 h when the THI increased from 56.2 to 73.8.

Consequently, cows dedicated less than 8 h per day to rest. With specific regard to air temperature and THI, ref. [101] observed that cows spend less time lying and more time standing when air temperature and THI increase. Lying time reduces by about 40 min when mean temperature is above 28 °C and by 48 min when the THI is higher than 72. These findings highlight the importance of monitoring cow behaviors and time spent lying and standing, these being measures useful indicators of cow adaptation to extreme environmental conditions and of their overall well-being.

The drinking behavior of dairy cows is closely linked to their ability to cope with high temperatures, as water consumption increases with increasing temperature due to the need for evaporative cooling [102,103,104]. Ref. [104] observed that moving from a THI equal to 57 to a THI equal to 72, lactating cows drank 21% more. The correlation between drinking behavior and THI is further supported by study [105] where free-range dairy cows had the most drinking episodes when the THI was between 78 and 82. Drinking patterns were assessed over a span of three consecutive days. Each hour of the day underwent systematic categorization based on the THI: “normal” (THI < 70), “alert” (70 < THI < 78), “danger” (79 < THI < 82), “emergency” (THI > 82). Incorporating a standardized stress index, which quantifies various environmental indicators, can provide an easily interpretable metric for assessing stress levels. This index can complement decision support system by offering a concise measure of environmental data to monitor stress indicators. Moreover, ref. [106] observed that drinking frequency increased consistently, i.e., from 0.12 to 0.23 l/THI, throughout a range of THI increasing from 51.4 to 79.7. Additionally, changes in drinking patterns were observed between different troughs based on the dominance hierarchy of the cows, with a seasonal correlation of drinking time with THI [107]. Likewise, study [108] evaluated the ways in which animal behaviors alter in a subtropical location throughout the hot season, with or without unrestricted access to shaded area. Cows having access to shaded area had lower water ingestion episodes than cows without access to shade. These results were confirmed in [109], which examined the effects of heat stress on the behavior at the drinker in 69 indoor-housed lactating Holstein dairy cows. The reviewed literature agrees on the fact that cows in heat stress consume a greater amount of water, spend more time at the drinker, approach the drinker more frequently, and engage in more competitive interactions at the drinker.

It is well known that heat stress leads dairy cows to reduce their daily feed intake, resulting in a decrease in milk yield and an increase in health issues. An indicator of heat stress in dairy cows is the decrease in appetite. Reduced feed intake is attributed to the discomfort caused by high temperature, as well as the energy required for sweat production, which is needed to regulate the cow’s body temperature [38,110]. In animals experiencing heat stress, a physiological adaptation to cope with internal metabolic heat production is the reduction in dry matter intake (DMI). In [54], significant differences were noticed in the feed sorting behavior of cows receiving evaporative cooling and those suffering from heat stress. Specifically cows under acute or chronic heat stress and without cooling opted for long ration particles, whereas cows receiving evaporative cooling did not sort for any proportion. Changes in behavior according to data gathered from the rumen environment may explain the consequences of heat stress [111]. Moreover, in the study conducted in ref. [62], cattle undergoing heat stress experienced a reduction in feed intake; this was observed in animals raised under thermoneutral conditions of 15% in the case of high-energy diet and 20% in the case of low-energy diet. Cows experiencing heat stress eat and ruminate less, which results in fewer buffering chemicals entering the rumen, where ruminating is the primary activator of saliva production [70]. According to [112], the mean daily DMI decreased less in heat-tolerant cows than in heat-susceptible cows. By decreasing the fermentation heat produced by the rumen, a reduction in daily DMI can be observed, which is a behavioral adaptation that decreases the generation of metabolic heat [113]. The additional metabolic heat production generated by dairy production may account for the more rapid adaptation observed in lactating cows [70,101]. Furthermore, a lack of DMI in transition dairy cows having higher energy need could result in an adverse energy balance [114]. Prior research has shown that heat stress can reduce de novo fatty acid synthesis in the mammary gland [115], which might cause acetate deficiency caused by a lower feed intake [116].

Animals have developed various strategies to cope with heat stress, and one of the most effective is to decrease the metabolic energy output through metabolic adaptation [117]. However, if heat stress persists for an extended period, it can lead to lethal heat stress. As previously mentioned, this condition can compromise the ability of the animals to thermoregulate, leading to hyperthermia, which can cause acute metabolic and systemic diseases [59]. The presence of hyperthermia and dehydration has been correlated with heightened neuromuscular fatigue and compromised movement coordination in animals. Hence, in regions characterized by hot climates, the potential for injury escalation is amplified [118]. Ref. [59] reported that lethal heat stress can result in a range of disorders, including endotoxemia, electrolyte imbalances, abnormalities of the acid-base system, and physiological disturbances including respiratory, cardiac, and renal failure. In vitro experimental models have demonstrated that a reducing tissue factor essentially limits inflammation-induced thrombin generation, thereby preventing endotoxemia [119]. However, the coagulation cascade can become widely activated by the tissue factor, generating thrombin, stimulating platelets, and producing platelet-–fibrin clots. Local perfusion consequences can lead to tissue hypoxia and organ failure caused by these microthrombi [120].

The conversion of feed into energy, for various metabolic activities such as milk production, maintenance, and development, is referred to as energetic metabolism in dairy cattle. Catabolism and anabolism are the two major processes involved in energetic metabolism. Under conditions of stress caused by high temperatures, lactating dairy cows experience a significant decrease in their blood glucose levels if compared to those maintained in thermal neutrality. This decrease primarily arises from a concomitant lower DMI, leading to a reduction in the overall glucose synthesis within the animal body [121,122]. Lactating cows that are subjected to heat stress exhibit high levels of plasma insulin, despite experiencing a decline in their blood glucose levels. This suggests that factors beyond blood glucose levels can impact the secretion of insulin in response to stressors [122,123]. Furthermore, several researchers have concluded that cows subject to heat stress show increased plasma urea nitrogen levels, indicating greater ammonia absorption in the gastrointestinal tract and enhanced hepatic elimination and amino acid deamination [124]. The results in [125] imply that the reduction in plasma amino acids measured during heat stress may be attributable to heightened amino acid consumption as gluconeogenic precursors. The occurrence of heat stress in lactating dairy cows has been shown to increase the quantity of hepatic glucose production [48]. With specific regard to the metabolism of proteins, ref. [126] revealed that heat stress led to the mobilization of skeletal muscle proteins, confirmed by an increase in blood creatine. These findings elucidate that short-term heat stress can enhance both adipose tissue lipolysis and skeletal muscle proteolysis as an energy source, whereas long-term heat stress inhibits adipose tissue lipolysis while intensifying skeletal muscle proteolysis.

Several studies have shown the negative correlation between milk production and THI [15,34,127,128], particularly when the daily average THI overcomes the value of 68 [17]. The findings in [129] further supported this relationship, with an increase in THI leading to a decrease in daily milk production, especially in high-productivity cows. Additionally, the level of activity and milk yield decrease in response to heat load, particularly three [130,131] or five days [132] after the exposure. Ref. [133] found that the highest average milk yield was obtained in October while the lowest yield was recorded in July, thereby confirming the inverse correlation between milk yield and heat stress effects modelled with the THI. Similarly, ref. [134] observed that dairy cattle exposed to high temperatures and reducing the feed intake had a considerable drop, of about 20%, in the milk production compared to the part of the dairy cattle reared in optimal climatic conditions. In [135], it was observed that cattle that started the milking period during the hotter season of the year exhibited the lowest milk production, whereas cows that started the milking period in intermediate seasons demonstrated the highest milk yield. A recent investigation carried out in [136] revealed that during the final stage of pregnancy, an increase in the maternal body temperature could lead to a reduction in the amount of milk yield up to the three following lactations. The stage of lactation is a crucial factor in determining the impact of heat stress experienced by animals, since mid-lactation cows exhibit greater susceptibility compared to cows in early and late lactation stages. Additionally, during the early lactation stage, milk production in cows is largely sustained by mobilization of tissue stores and in a lower extent by feed intake. Conversely, in the mid-lactation stage, milk yield primarily relies on feed intake [137]. In this regard, past experimental tests have investigated the impact of reducing consumption of DMI on milk yield in animals affected by heat stress. It can be speculated that the direct influence of heat stress on milk yield indicates that a percentage ranging from 35% [138] to 50% [122] of the overall productivity drop can be attributed to a reduction in DMI [139]. Due to the higher efficiency of metabolic tissue store utilization in comparison with the metabolic utilization of feed, it is expected that early lactating cows would generate a lesser amount of metabolic heat per unit of produced milk with respect to mid-lactating cows [137]. This agrees with [139], the authors of which investigated cows in an intermediate phase of lactation, and reported an increased vulnerability to the adverse conditions associated to high THI recording modification of milk composition and a decline in milk production. Likewise, heat stress during the dry period undermines the development of the mammary gland prior to giving birth, leading to a decrease in milk production during the following lactation [140]. The thermal stress not only impacts milk quantity, but also adversely affects milk quality, leading to a decreased levels of proteins, fats and lactose [52].

Several investigations [49,141,142] have confirmed the strong correlation between THI and percentage of both milk fat and milk protein. Ref. [144] noted a reduction in the concentration of fat and protein in milk during the summer months, whereas an increase in concentration was observed during the winter season, but on the other hand, it was found that lactose remained constant across the seasons. Contrary to this finding, in work [145], it was observed that the lactose level in grazing cattle decreases during the summer season compared to the winter season. Lactose is linked to the production of α-La, a coenzyme necessary for its formation. This specific whey protein is synthesized in the mammary gland, where α-La interacts with β-1,4-galactosyltransferase in the Golgi apparatus of the mammary gland epithelial cells to facilitate the formation of the lactose–synthase enzyme. As stated in [146], α-La alters the substrate specificity of β-1,4-galactosyltransferase to enable the creation of lactose from glucose and UDP-galactose.

With specific regard to milk fat, lipid represents a fundamental constituent, with a substantial portion consisting of polar lipids that act as emulsifiers and serve as the primary structural elements of the fat globule membrane. They are essential for the milk emulsion system to remain stable [147]. In [115], the content of polar lipids underwent a reduction of 43% following the exposure to heat stress for a duration of two days, subsequently decreasing further to 52% after four days from the day of the exposure. Following a five-day period of recovery, the polar lipid content rebounded to its baseline level. Furthermore, ref. [148] observed that cows who were exposed to high temperatures for one or more consecutive days produced less milk fat compared to animals that were not under heat stress conditions. With regard to the content of proteins in milk, the work revealed that the cows need to be stressed for at least three consecutive days to observe a significant reduction in protein content. There are various possible explanations for the decrease in milk protein, some of which may be inherit to the mammary gland. Ref. [134] revealed that the decrease in milk protein observed in heat stressed cows occurs due to a specific downregulation of protein synthesis in the mammary gland and not a byproduct of a general decrease in milk production. The authors determined that heat stress exclusively influenced the individual casein mass fraction proportions, leading to an increase in α_S1-casein and a decrease in α_S2-casein. Following [125], the authors of which investigated the impact of high temperatures on the generation of milk protein, cattle exposed to heat stress displayed a decrease in the total amount of amino acids in their plasma, which was lowered by 17.1%. Both essential and non-essential amino acids were impacted by this reduction.

The immune response of dairy cattle is a complex process involving various cellular and molecular mechanisms that are affected by changes in environmental conditions. For instance, heat stress has been proven to decrease the activity and quantity of specific immune cells, such as neutrophils and lymphocytes. According to [148], heat stress is linked to a decrease in in vitro neutrophil phagocytosis and lymphocyte proliferation in crossbred cattle. These cells perform critical functions in battling infections and illness, and their decreased activity can make dairy cattle more susceptible to infectious pathogens. Likewise, the presence of heat stress causes a significant decrease in the percentage of CD21+MHCII+ B cell populations in both Holstein and Jersey steers. Furthermore, high temperature decreases the level of myeloperoxidase in polymorphonuclear cells in Holstein steers, whereas heat stress decreases the WC1 + γδ T cell populations in Jersey steers. Sahiwai, in comparison to Gir and Tharparkar breeds, exhibit elevated heat stress indicators, including reduced phagocytic activity, increased somatic cell count, heightened neutrophil population, elevated milk cortisol levels, and the activation of heat shock and cell adhesion markers [149]. In the mammary gland, neutrophils perform the critical role of phagocytosis and intracellular killing. This involves the process of chemoattraction, where neutrophils are drawn towards bacteria, followed by their engulfment using two distinct mechanisms, namely the respiratory burst and digestion using lysosomal enzymes [150]. In addition to the influence on immune cell function, heat stress may affect the synthesis of immune system elements such as cytokines and antibodies. Temperature increases has been demonstrated to reduce the production of cytokines, which are critical in immune response coordination. In [151], it is shown that heat stressed cows have reduced blood plasma concentrations of inflammatory cytokines and immunoglobulins (i.e., IgA, IgM, and IgG). As a result, the capacity of the immune system to mount a successful defense against pathogens may be compromised. According to these results, providing dairy cows with cooling measures during the initial period of the lactation has a positive impact on their production performance, metabolic indicators, immune response, and antioxidant capacity. Exposure to a heat stress environment alters physiological traits, which might also interact with bacterial metabolism and modify the systemic levels of cytokines, impacting the function of the brain–gut axis in dairy cows [152]. In comparison to cows maintained in thermoneutral conditions, ref. [153] reports that heat stressed cows express more IL-10. Similarly, ref. [154] notes that heat stress considerably affects the levels of IL-6 in the blood of cows during the interval preceding calving. These levels are much higher in heat-stressed animals compared to those in cows raised in non-stressing climate conditions. Furthermore, exposure to high temperatures can result in oxidative stress, a state in which the capability of the body to neutralize reactive oxygen species is out of balance with the amount of these species produced [59]. Oxidative stress may harm cells and tissues, as well as affect immune function by lowering immune cells capacity to operate in a proper way. In presence of a high THI level, mitochondria produce more superoxide anion. Additionally, the stress leads to an increase in the production of transition metal ions, which boosts the creation of superoxide anion [59]. Oxidative stress in dairy cows is caused by excessive reactive oxygen species (ROS) production and consequent mitochondrial dysfunction resulting from high temperatures. This oxidative stress, promoting both insulin resistance and apoptosis, which are negatively associated with milk protein synthesis, leads to a reduction in total milk protein [155]. Heat stress increases the risk of various diseases in dairy cattle, such as mastitis and lameness. High temperatures can weaken the immune system and make it more vulnerable to infections. Several studies have also evaluated the consequences of seasonal variations on the immunity of dairy cows. Based on a 12-month study conducted across 13 different herds, ref. [156] finds that the prevalence of Streptococcus dysgalactiae infections is lower at the beginning of the housing period compared to the pasture season [156]. Further, the impact of THI on female reproduction and health characteristics such as clinical mastitis, retained placenta, and puerperal disorders, is evaluated in 22,212 cows from Day 0 to Day 10 postpartum in [157], establishing that that the incidences rise along with a rising average THI. The immunological status responses of dairy cattle to heat stress involves several complicated pathways that are currently unknown. Nonetheless, it is known that heat stress can have a deleterious impact on dairy cattle’s immune systems, making them more prone to infections and illness.