Ruminants

Feeding a balanced diet such as total mixed ration (TMR) is a widely adopted feeding strategy providing a uniformly blended diet of roughages, concentrates, and supplements that enhances ruminant productivity by optimizing nutrient utilization, stabilizing rumen fermentation, and improving microbial activity. Scientific studies have confirmed that TMR increases dry matter intake (DMI), milk yield, and growth performance in dairy and beef cattle, as well as in sheep and goats. TMR’s advantages include consistent feed quality, reduced selective feeding, and improved feed efficiency. A key benefit of TMR is its ability to promote the production of volatile fatty acids (VFAs), which are the primary energy source for ruminants, particularly propionate. This enhances energy metabolism, resulting in higher carcass yields, increased milk production, and economic benefits compared to conventional or supplementary feeding systems. However, TMR feeding is also susceptible to mycotoxin contamination (e.g., aflatoxins, zearalenone), potential effects on methane emissions, and the need for precise formulation to maintain consistency and optimise profitability. Prevention and good practices, including routine inspection of feed for pathogens and vulnerable ingredients, as well as careful management of particle size and forage-to-concentrate ratios, are crucial in preventing subacute ruminal acidosis (SARA) and the development of other subclinical diseases. Mycotoxin binders, such as hydrated sodium calcium aluminosilicate, can also reduce mycotoxin absorption. Another advantage of practicing TMR is that it can support sustainable farming by integrating agro-industrial byproducts, which minimises environmental impact. In conclusion, TMR is a widely adopted feeding strategy that significantly enhances ruminant productivity by optimizing nutrient utilization, stabilizing rumen fermentation, and improving microbial activity, leading to increased dry matter intake, milk yield, and growth performance. It offers key benefits such as consistent feed quality, reduced selective feeding, improved feed efficiency, and enhanced energy metabolism, providing economic advantages and supporting sustainable farming through agro-industrial byproduct integration. However, its implementation requires careful management to mitigate risks, including mycotoxin contamination, potential impacts on methane emissions, and digestive issues like SARA if formulation is not precise. Therefore, for sustainable production, future research should focus on optimizing TMR formulations with alternative ingredients (e.g., agro-industrial byproducts) and precision feeding strategies to enhance livestock health and animal productivity while minimizing environmental impacts. Full article

This study aimed to evaluate the effects of age and sex on carcass traits, commercial cuts, and meat quality in crossbred kids. A total of 32 Boer × native crossbred kids were used, equally distributed according to sex and slaughter age: 16 male kids (8 slaughtered at 70 days and 8 at 100 days) and 16 female kids (8 at 70 days and 8 at 100 days). However, slaughter age significantly affected (p < 0.05) total weight gain, empty body weight, hot and cold carcass weights, loin eye area, internal fat percentage, and carcass compactness index. A significant interaction (p < 0.05) between sex and age was found for thoracic depth. In addition, leg length and leg compactness index were significantly influenced (p < 0.05) by both factors. Slaughter age also had a significant effect (p < 0.05) on internal body length, leg length, rump width, thigh perimeter, rump perimeter, chest circumference, thoracic depth, internal thoracic depth, and carcass compactness index. Regarding commercial cuts (neck, shoulder, ribs, loin, and leg), slaughter age had a significant influence (p < 0.05), reflecting the same pattern observed in hot and cold carcass weights, and indicating consistent distribution across cuts. In conclusion, slaughter age had a more pronounced effect on carcass characteristics than sex, highlighting its importance in determining meat yield and quality in crossbred kids. Full article

Selection indices enhance dairy cattle breeding by optimizing multiple traits simultaneously. This study analyzed 2181 lactation records from Holstein–Friesian cows at Sakha Experimental Farm, Egypt, to evaluate selection indices for 305-day milk yield (MY), fat yield (FY), and protein yield (PY) using two relative economic value (REV) methods: actual economic values (REV1) and one phenotypic standard deviation (REV2). Using MTDFREML software, we estimated heritabilities of 0.27 ± 0.01 (MY), 0.22 ± 0.01 (FY), and 0.28 ± 0.02 (PY). Four selection indices were constructed based on actual relative economic values (REV1) and one phenotypic standard deviation (REV2). The comprehensive index (I₁) incorporated all three key production traits, viz., MY, FY, and PY, to maximize the genetic merit of the aggregate genotype. In contrast, the reduced indices (I₂, I₃, and I₄) included only two traits each. The I₂ incorporated MY and FY, the I₃ included MY and PY, and the I₄ included FY and PY. The index I₁ (including all traits) yields the highest genetic gains (305 kg MY, 14.0 kg FY, 11.93 kg PY per generation). Both REV methods produced comparable genetic gains, but REV2 is recommended for its computational simplicity. These findings support the use of selection indices for genetic improvement in Holstein–Friesian cows, offering practical guidance for dairy breeding programs in Egypt and similar environments. Full article

This study aims to analyze the technical and economic infrastructure of sheep farming enterprises operating in Türkiye. It assesses the demographic characteristics of enterprise owners, enterprise scales, production objectives, marketing strategies, and economic performance. Primary data were collected through face-to-face surveys conducted with 201 sheep farming enterprises during the 2023 production period. The sample was selected based on information provided by the Provincial Directorates of Agriculture and Forestry and the Breeding Sheep and Goat Breeders’ Associations. Data analysis was performed using SPSS 27. Categorical data related to enterprise characteristics and the demographic profiles of enterprise owners were examined. The findings indicate that the majority of enterprise owners are middle-aged or older individuals, have a low level of education, and operate predominantly within an extensive production system. The producers’ marketing methods rely mainly on direct sales. In conclusion, ensuring the sustainability of the sheep farming sector requires encouraging young producers to enter the industry, expanding educational programs, and adopting modern production techniques. Full article

MFD is a nutritional disorder in dairy cattle characterized by a reduction in milk fat content despite a normal or increased milk yield. This review synthesizes current knowledge on the biological mechanisms and nutritional factors contributing to the development of this condition. Disruptions in rumen fermentation and alterations in fatty acid biohydrogenation (particularly the formation of trans-10 fatty acids) are recognized as central contributors to MFD. Several theories have been proposed to explain its pathophysiology, including the glucogenic, volatile fatty acid, trans fatty acid, and biohydrogenation theories. MFD is most commonly associated with diets low in fiber and high in polyunsaturated fatty acids or starch, which promote the accumulation of fatty acid intermediates that inhibit mammary lipogenesis. Among these, trans-10, cis-12 conjugated linoleic acid is particularly notable for its potent suppression of de novo fatty acid synthesis in the mammary gland. While proper dietary formulation remains the most effective preventive strategy, nutritional interventions such as magnesium-based alkalinizers, sodium bicarbonate, intravenous arginine, and vitamin E have shown promise in mitigating established cases. This review underscores the importance of nutritional management in preserving milk fat synthesis and promoting overall animal health. Full article

The success of optimal ruminant production relies heavily on feed efficiency to deliver the necessary nutrients to animals. Nutritional deficiencies in livestock pose a significant challenge in regions experiencing prolonged fluctuations in resource availability and quality. In this context, the present study aimed to investigate the cumulative gas production (CGP) and in vitro degradability of silages made from spineless forage cactus (a native species) combined with high-fiber ingredients, to evaluate their viability as a sustainable, low-cost alternative to animal feed. The experiment involved ensiling spineless cactus genotypes with varying levels of sugarcane bagasse (0, 150, 300, 450, and 600 g/kg of dry matter) and a 1% urea–ammonium sulfate solution. The results indicated that for all genotypes studied, the CGP curves from silage composed solely of forage cactus differed significantly from those containing bagasse, which exhibited an initial phase characterized by little or no gas production. In vitro degradability was negatively influenced by the inclusion of bagasse at any level, resulting in decreased dry matter and organic matter degradability, as well as reduced CGP with increasing bagasse concentration. Therefore, the study demonstrated that the proposed combination of ingredients represents a promising sustainable feed supplement to enhance animal nutrition. Silage containing 150 g/kg of bagasse treated with urea offers a favorable balance between the energy required by rumen microflora and the benefits of fiber presence. Full article

Ruminant livestock production plays a crucial role in the agricultural systems of Sub-Saharan Africa, significantly supporting rural livelihoods through income generation, improved nutrition, and employment opportunities. Despite its importance, the sector continues to face substantial challenges, such as low feed quality, seasonal feed shortages, and climate-related stresses, all of which limit productivity and sustainability. Considering these challenges, the adoption of natural feed additives has emerged as a promising strategy to enhance animal performance, optimise nutrient utilisation, and mitigate environmental impacts, including the reduction of enteric methane emissions. This review underscores the significant potential of natural feed additives such as plant extracts, essential oils, probiotics, and mineral-based supplements such as fossil shell flour as sustainable alternatives to conventional growth promoters in ruminant production systems across the region. All available documented evidence on the topic from 2000 to 2024 was collated and synthesised through standardised methods of systematic review protocol—PRISMA. Out of 319 research papers downloaded, six were included and analysed directly or indirectly in this study. The results show that the addition of feed additives to ruminant diets in all the studies reviewed significantly (p < 0.05) improved growth parameters such as average daily growth (ADG), feed intake, and feed conversion ratio (FCR) compared to the control group. However, no significant (p > 0.05) effect was found on cold carcass weight (CCW), meat percentage, fat percentage, bone percentage, or intramuscular fat (IMF%) compared to the control. The available evidence indicates that these additives can provide tangible benefits, including improved growth performance, better feed efficiency, enhanced immune responses, and superior meat quality, while also supporting environmental sustainability by reducing nitrogen excretion and decreasing dependence on antimicrobial agents. Full article

Due to the feeding system (high-concentrate diet) during the early lactation stage, ruminal pH in dairy cows follows a diurnal pattern and can remain below the critical level of 5.5 for extended periods of the day. This study aimed to evaluate the effect of low ruminal pH on blood concentrations of certain macrominerals, trace minerals, and fat-soluble vitamins and on the oxidative status of dairy cows during the first half of lactation. Fifty-three randomly selected lactating Holstein cows were used; blood and ruminal fluid samples were collected from all cows on days 30, 90 and 150 of lactation. Blood samples were obtained via coccygeal venipuncture, while the ruminal fluid was obtained by rumenocentesis and the pH was measured immediately after collection. Using a threshold pH of 5.5, samples were classified as normal (pH > 5.5) or low pH (pH ≤ 5.5). Serum concentrations of Ca, Mg, K, Cr, Mn, Zn, Se, and vitamins A, D3, E, and K were not significantly affected by ruminal pH, either by days in milk or by their interaction (p > 0.05). Plasma malondialdehyde and reduced glutathione followed the same trend (p > 0.05). Copper concentration was significantly higher (p < 0.05), and Fe concentration tended to be higher in cows with low pH compared to those with normal pH (p = 0.052). On day 150 of lactation, Cu, Fe, and Co concentrations were significantly higher in low-pH cows compared to normal-pH cows (p < 0.05). Low ruminal pH is associated with significant changes in serum concentrations of copper, iron, and cobalt but has no significant effect on the oxidative status of the animals or on the serum concentrations of the macro elements and fat-soluble vitamins studied. Full article

The objective of this analysis was to compare the greenhouse gas (GHG) emissions from contemporary grazing cattle production with bison grazing, both modern and historical. The data sets used in this analysis were derived from existing research and conservation properties located outside of Manhattan, KS (USA), which are home to stocker cattle, cow–calf production (CCS), and grazing bison. For stocker cattle, 10 years of animal production data (2007–2016) from season-long stocking (SLS, grazing 156 d) and intensive early stocking systems (IES; 76 grazing d and 2× stocking density) were used for GHG calculations. Enteric CH₄, manure CH₄, and direct nitrous oxide emissions were estimated using the IPCC tier 2 methodology. Historic bison (HGB) enteric CH₄ estimates were calculated using a stocking density of 0.15 ha/animal and assuming that only 13% of grassland was used by bison each year. Within contemporary systems, IES had the lowest emissions (463.3 kg CO₂-eq./ha/yr), while SLS, CCS, and MGB had the highest estimates (494.7, 493.9, and 595.9 kg CO₂-eq./ha/yr, respectively). HGB had the lowest estimated annual emissions at 295.7 kg CO₂-eq./ha/yr. These results imply that the historic grazing baseline of this grassland system is lower but similar to that of contemporary grazing cattle in the Great Plains region. Full article

Geometric morphometrics is performed on different species in different contexts. Here, the aim was to investigate morphological differences in the head of the Sanmartinero Creole bovine to examine head shape variations between sexes using geometric morphometric methods. A sample of cranial pictures of 43 animals (13 males and 30 females) was obtained, and form (size + shape) was studied by means of geometric morphometric techniques using a set of 14 landmarks. This approach eliminated potential dietary effects, ensuring that the observed shape variations were primarily due to intrinsic morphological differences. Sexual dimorphism was found in form (for both size and shape) of the head of the Sanmartinero Creole bovine breed. Males had significantly larger heads based on centroid size (U = 714, p = 0.0004), confirming true sexual size differences, and Principal Component Analysis revealed overlapping head shapes with sexual dimorphism concentrated at midline sagittal landmarks (between the most rostral and caudal orbit points) and paired lateral points, indicating that males have broader and longer heads. The two evaluated characters (head size and shape) are of special interest for the conservation of the breed, especially in those cases whose objectives are to maintain the uniqueness, distinctiveness, and uniformity of the populations. This study analyzed animals subjected to the same feeding program, ensuring the elimination of additional variables. Full article

The aims of the current study were to characterize the natural compounds of Salsola tragus via GC-MS and determine its effects as a forage source on lamb growth, in vitro rumen fermentation kinetics, carcass quality, cellular damage, and metabolomic meat interaction. Twenty-one Rambouillet lambs were randomly assigned to one of three experimental diets (seven lambs per treatment): (1) a control diet (W/o-Salsola) containing 300 g/kg dry matter (DM) of sorghum stover; (2) a diet with a medium inclusion of Salsola tragus (15-Salsola), which contained 150 g/kg DM of Salsola tragus and 150 g/kg DM of sorghum stover; and (3) a diet comprising 300 g/kg of Salsola tragus (30-Salsola). The results showed there were no differences (p > 0.05) in lamb growth performance during the fattening phase. The in vitro gas analysis demonstrated that the 30-Salsola treatment increased lag time h⁻¹ (p < 0.05) and reduced gas production (p = 0.03). The metabolomic analysis findings suggest that the treatments that included Salsola tragus significantly positively affect the metabolomic composition of meat (p < 0.05). The use of 15-Salsola as a source of forage is promising for feeding lambs during the fattening phase. Full article

A key factor in calculating dairy cows’ nitrogen (N) excretion is knowing the amount of daily excreted urine. The present study aimed to investigate two methods to calculate the daily urine volume (UV) excreted using spot urine samples. Data were obtained from nine balance experiments involving 47 lactating and seven non-lactating German Holstein cows, with an average body weight (BW) of 620 ± 95 kg and an average age of 5.6 ± 1.4 years. Daily urinary creatinine (Cr) and UVs were known for all animals. The first method was developed by linearly regressing the daily excreted amount of Cr in urine against BW (p < 0.001; R² = 0.51; RSE: 2.8). The slope of the regression was used to calculate UV. The second method includes a non-linear regression of UV on Cr concentration in urine, allowing direct estimation of UV without knowledge of BW (p < 0.001; RSE: 8.13). Both estimation methods were compared to the standard method to determine UV from balance trials using Lin’s concordance correlation coefficient (CCC) and Bland–Altman plots. The first method had a CCC of 0.81, and the second method had a CCC of 0.85. Both methods can confidently be applied to calculate UV. Therefore, the second method is usable if BW is unavailable. Full article

Overmilking occurs when the teat cups remain attached to the udder during milking, even though there is little or no milk flow. This puts pressure on the teat tissue and reduces milk production due to longer milking times, meaning fewer cows are milked per hour. Therefore, the correct removal of the teat cup at the end of mechanical milking is crucial for the milking process. The aim of this study was to describe the factors influencing automatic teat cup removal (ATCR) at the end of mechanical milking and to demonstrate its importance for udder health, milk production and milk quality. There are considerable differences between milking system suppliers and countries regarding the minimum removal of the teat cup at the end of the milking process. However, to ensure good milk quality, prevent teat damage and reduce the risk of mastitis, it is important to shorten the working time of the milking machine on the udder in both automatic and conventional milking systems. For this reason, several studies have shown that increasing the milk flow switch point effectively reduces milking time, especially in automatic milking systems where dairy cows are milked more than twice a day. However, when the ATCR setting was increased above 0.5 kg·min⁻¹, milk production decreased, and the number of somatic cells in the milk produced increased. Therefore, the use of ATCR at a milk flow rate of 0.2 kg·min⁻¹ significantly increased milk production, improved milk quality and maintained udder health when a low vacuum level (34–36 kPa) was used in milking machines such as MultiLactor and StimuLactor (Siliconform, Germany). In conclusion, ATCR at a milk flow of 0.2–0.3 kg·min⁻¹ is a useful level to achieve various goals on dairy farms when a low vacuum of 34–36 is used in the milking machine. If the milking machine uses a higher vacuum, it is possible to program a higher ATCR at a milk flow of up to 0.5 kg·min⁻¹. Full article

The 8th revised edition of the Nutrient Requirements of Beef Cattle was released in 2016, with the recommendations provided in the publication being used extensively in both research and production settings. In the context of research needs identified in that publication, our objective was to review research on beef cattle nutrient requirements published since 2016 and identify knowledge gaps that should be addressed. Relative to energy requirements, the effects of environmental temperature and grazing activity, along with stress and disease, on maintenance requirements are inadequately characterized or defined. In addition, relationships between retained energy and protein should be more fully elucidated, and additional guidance on body weight at a target compositional endpoint is needed. Areas of continuing concern include accurately and precisely predicting microbial protein supply, predicting N recycling, and the metabolizable protein requirements for maintenance. Mineral and vitamin requirements are often challenging because of a lack of consistency in models used to determine requirements and potential effects of unique production settings on requirements. Based on recent research with feedlot cattle, zinc and chromium requirements should be examined more closely. Because predictions of dry matter intake are critical to supplying nutrients, additional development of prediction equations is needed, especially for beef cows and grazing beef cattle in general. Given considerable research in prediction of greenhouse gases, reevaluation of 2016 recommendations is warranted, along with a need for the updating of equations to predict excretions of N and P. Composition of feeds, particularly byproducts from ethanol production or other industrial streams, represents a knowledge gap, with obtaining reliable energy values of these feeds being a notable challenge. Nutritional models provide the means to integrate nutrient requirement recommendations into practice, and moving towards mechanistic models that take advantage of artificial intelligence and precision livestock farming technologies will be critical to developing future modeling systems. Full article

This study aimed to evaluate the microbiological quality of colostrum on three dairy farms with different colostrum management hygiene practices and to compare it with the current colostrum quality guidelines. On farm A, colostrum was fed raw, while on farms B and C it was heat treated. On farms A and B, the feeding equipment was cleaned manually, while on farm C, an automated cleaning system was used. Samples were collected from the calf-feeding equipment and submitted for microbial culture: total plate count (TPC); total coliform count (TCC); and E. coli, enterobacteria (ENTB), staphylococci (STAP), and lactic acid bacteria counts. In addition, pH, water activity (a_W), and Brix were analyzed. Colostrum quality was defined as follows: good quality (GQ)—TPC < 100,000, TCC < 10,000, STAP < 50,000 cfu/mL, and Brix ≥ 22%; excellent quality (EQ)—TPC < 20,000, TCC < 100, STAP < 5000 cfu/mL, and Brix ≥ 25%. Mean concentrations were as follows: TPC was 3.99 × 10⁵ cfu/mL (min: 40.00, max: 1.32 × 10⁷ cfu/mL); TCC was 1.17 × 10⁴ cfu/mL (min: <detection limit, max: 6.37 × 10⁵ cfu/mL); and STAP was 1.77 × 10⁴ cfu/mL (min: <detection limit, max: 3.50 × 10⁵ cfu/mL). Approximately 54% (GQ) and 32% (EQ) of samples met the defined criteria. Farm C consistently showed lower microbial counts across all culture types. Colostrum from farm B had lower TCC, LAB, and E. coli counts than farm A but not TPC, STAP, and ENTB. These results showed that a considerable proportion of calves were fed colostrum with suboptimal quality, especially when less rigorous hygiene practices were implemented. Full article

Understanding the genetic and phenotypic basis of economically important traits is essential for designing effective breeding programs in livestock. This study aimed to evaluate the phenotypic performance and estimate genetic parameters for one reproductive trait—litter size at birth (LSB)—and three pre-weaning growth traits—birth weight (BW), weaning weight (WW), and average daily gain (ADG)—in a population of 1888 Zaraibi kids born between 2018 and 2023. Genetic parameters were estimated using animal models implemented in the MTDFREML software. The overall least squares means (±standard error) for LSB, BW, WW, and ADG were 2.22 ± 0.02, 2.03 ± 0.01 kg, 10.22 ± 0.05 kg, and 90.00 ± 0.50 g/day, respectively. Statistical analyses indicated that month of birth, year of birth, and type of birth had significant (p < 0.05) effects on all traits, while the sex of the kids had no significant effect (p > 0.05) on LSB. Direct heritability estimates (h²_d) derived from Model 1 (including additive genetic, permanent environmental, and residual effects) were 0.13 ± 0.01 for LSB, 0.30 ± 0.04 for BW, 0.38 ± 0.01 for WW, and 0.30 ± 0.10 for ADG. Under Model 2 (which incorporated maternal genetic effects and their covariance with direct genetic effects), maternal heritability (h²_m) estimates for LSB, BW, WW, and ADG were 0.05 ± 0.01, 0.15 ± 0.01, 0.12 ± 0.01, and 0.14 ± 0.01, respectively. Overall, the results emphasize the importance of maternal genetic effects in influencing pre-weaning growth traits. Therefore, maternal genetic components should be explicitly considered in genetic evaluation and selection strategies aimed at improving early growth performance in Zaraibi goats. Full article

The conservation of local breeds plays a strategic role in maintaining genetic variability, ensuring adaptive responses to environmental challenges, and preserving the cultural and socioeconomic structures of traditional communities. In this context, this study explores the potential disappearance of traditional knowledge about local breeds from an ethnozootechnical perspective. The objectives were (I) to establish the breeding history of goat and sheep breeds/ecotypes in the semi-arid region of Paraíba; (II) to estimate the diversity index; and (III) to evaluate the selection criteria used by local communities in four territories: Coletivo, Borborema, Folia, and Casaco. The study aims to support genetic conservation and improvement programs. Data collection was participatory, involving breeders from all territories. To recover the breeds’ history, questionnaires were applied to the oldest breeders, called the “guardians.” Two workshops were held to assess the diversity of breeds in the past landscape (PP) and current landscape (PA), using the Recall technique. Responses were recorded in spreadsheets for analysis. Descriptive statistics and multiple correspondence analysis (MCA) were used to assess animal distribution. The Shannon index indicated a drop in goat breed diversity, from 1.3 (PP) to 0.87 (PA). For sheep breeds, it decreased slightly from 0.7 to 0.66. Breeders reported valuing traits such as adaptability, disease resistance, fertility, and conformation. Their strong emotional connection with the animals highlights the breeds’ cultural relevance. A strong connection was found between the loss of genetic material in the studied territories and the extinction of local communities’ knowledge about local breeds. Full article

Cytokine and Anti-Müllerian Hormone (AMH) profiling were performed in prepubertal Murrah buffalo heifers’ post-progesterone supplementation and Co-synchronization (Co-synch) protocol. Cytokine (IFNγ, IL6, IL1, IL13, TNFα, and TGFβ), AMH, progesterone, and estrogen hormone estimations were conducted. The ovarian follicular pattern and fertility outcome were recorded. Eighteen prepubertal heifers, 15–17 months of age, 250–300 kg body weight (BWt) were randomly divided into three groups, Group 1: n = 6, supplemented with altrenogest 0.044 mg/kg BWt/day/heifer orally for 14 days along with Co-synch program; Group 2: n = 6, implanted Controlled Internal Drug Release (CIDR) for 14 days and co-synch program, Group 3: n = 6, received Co-synch program. Ultrasonography was performed to determine ovarian follicle status on respective days of sampling. Pregnancy diagnosis was performed around 45 and 60 days post artificial insemination (AI). The Estradiol 17-β level remained constant in Group 1, Group 2, and Group 3 during the progesterone treatment and Co-synch treatment. Estradiol levels during Co-synch were significantly less (p = 0.024) on day 9 of Co-synch (14.41 ± 1.97 pg/mL) than on day 0 (20.11 ± 0.36 pg/mL) and on day 7 (19.77 ± 0.34 pg/mL) in prepubertal buffalo heifers in Group 1. However, no significance was observed in other groups. Progesterone levels in buffalo heifers subjected to synchronization protocols varied significantly (p < 0.05) on day 7 of progesterone (P4) treatment between Group 1, Group 2, and Group 3. Supplementation of altrenogest showed a significant (p = 0.043) increase in progesterone levels by day 14 of altrenogest treatment. Progesterone varied significantly in all groups on day 9 of Co-synch protocol [Group 1 (p = 0.020), Group 2 (p = 0.041), and Group 3 (p = 0.007)]. Cytokine IFNγ showed high correlation with progesterone, indicating the role of IFNγ in puberty in buffalo heifers (r = 0.626, p < 0.01). Anti-Müllerian Hormone had a significant positive correlation when supplemented with altrenogest with IFNγ (r = 0.673, p < 0.01) and TGFβ (r = 0.463, p < 0.01), whereas it was a negatively correlated with TNFα (r = −0.34, p < 0.05). Based on Karl Pearson correlation coefficients, IL13 and TGFβ could be considered as markers for puberty in buffalo heifers. Full article