Open Veterinary Journal, (2026), Vol. 16(4):2155-2163
Research Article
10.5455/OVJ.2026.v16.i4.18
Assessment of health conditions in Khmer native × Hariana crossbred cattle under a free-grazing system
Sophany Morm1*, Mach Din1, Pao Srean1, Jinhu Huang2, Jinxin Liu3, Vey Seb4 ,
Kheng Kheak4, Phiny Chiv5, Areerat Lunpha6 and Ruangyote Pilajun6
1Department of Animal Science, Faculty of Agriculture and Food Processing, National University of Battambang, Battambang, Cambodia
2College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China
3College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, China
4Graduate School, National University of Battambang, Battambang, Cambodia
5Research and Development Office, Faculty of Agriculture, Svay Rieng University, Svay Rieng, Cambodia
6Department of Animal Science, Faculty of Agriculture, Ubon Ratchathani University, Ubon Ratchathani, Thailand
*Corresponding Author: Sophany Morm. Department of Animal Science, Faculty of Agriculture and Food Processing, National University of Battambang, Battambang, Cambodia. Email: sophanymorm [at] gmail.com
Submitted: 31/12/2025 Revised: 11/03/2026 Accepted: 29/03/2026 Published: 30/04/2026
© 2025 Open Veterinary Journal
This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way.
Abstract
Background: Cattle play an important role in supporting food security and rural livelihoods economy in Cambodia. However, cattle production systems in Cambodia remain largely traditional, characterized by extensive grazing, limited feed supplementation, low genetic potential, and inadequate health management, which result in low productivity and increased vulnerability to diseases. Therefore, ensuring sustainable productivity in the cattle sector in Cambodia requires a focus on health management, which is essential to enhance animal health resilience and reduce disease vulnerability.
Aim: This study aimed to evaluate the condition of Khmer native × Hariana crossbred cattle under a free-grazing system, with a focus on differences between males and females.
Methods: Ten cattle were selected, consisting of five males (206.80 ± 10.15 kg/head) and five females (192.60 ± 12.59 kg/head) based on their initial body weight (IBW). The dietary treatments were arranged in a Completely Randomized Design (CRD) with two treatments and five replications, each replication containing one animal. The feeding treatments were as follows: FGm, free-grazing (male), and FGf, free-grazing (female).
Results: The results indeed showed that the IBW and final body weight (FBW) did not differ significantly between the groups (p > 0.05). Eimeria spp. egg per gram (EPG) counts were significantly different between groups during the first sampling (p < 0.05), but no significant difference was observed in the second sampling (p > 0.05). Glucose and triglyceride concentrations were lower than the laboratory reference values, while creatinine and blood urea nitrogen levels were higher; however, none of these biochemical parameters differed significantly between groups (p > 0.05).
Conclusion: These findings suggest that free-grazing management may adversely affect cattle health. Thus, to ensure clear and measurable results, data collection should be conducted over a longer period in future research on native crossbred beef cattle.
Keywords: Free-grazing, Hariana crossbred cattle, Health assessment, Khmer native cattle.
Introduction
Bos indicus weighing an average of 250–300 kg and measures approximately 1.5–1.7 meters in body length. It typically has yellow, maroon, or red coat coloration and short neck. This species is highly resilient and can survive on poor-quality feed and disease. However, Hariana cattle originated from India and were introduced to Cambodia between 1960s and 1980s. These cattle are known for their disease resistance (Kumar et al., 2019). In addition, this variety is well recognized for its disease resistance (Kumar et al., 2019). Nevertheless, Khmer native x Hariana crossbred cattle are primarily raised for beef production. Crossbred cattle are widely reared due to their adaptability to the tropical monsoon climate, enhanced disease resistant, and strong alignment with local market demand. Nevertheless, crossbreeding must be well planned, carefully controlled, and supported by proper management and conservation of local breeds to ensure long-term sustainability. Cattle-raising systems are particularly important, serving as a major source of household income and contributing to national economic assets (Ashley et al., 2018). In rural areas, farmers show strong interest in animal husbandry; however, they continue to face various technical limitations, including shortages of feed and water resources, operational constraints, breeding challenges, and the influence of diseases. Furthermore, animal feed resources need improving (Al-Baadani et al., 2022; Morm et al., 2024b). Most livestock farmers cultivate commercial grasses to meet the nutritional needs of ruminants and support time-efficient management, but this practice requires large areas of land (Cantalapiedra-Hijar et al., 2018). Currently, cattle production in Cambodia faces numerous constraints, including insufficient forage quantity and quality, internal and external parasitism, and other disease-related challenges that reduce overall productivity (Darith et al., 2017). These constraints convey to waste of time and saving diseases of owner livestock investigator (MAFF , 2021). Improving livestock feeding systems is therefore necessary to enhance overall animal health. Free-grazing is most popular for cattle raising, but this cause impacts cattle health due to insufficient feed and other impacted sources of cattle health (Kijlstra et al., 2009; Morm et al., 2024a). Thus, intensity grazing stability is an optional level of a productive state, which plays an important role in enhancing metabolizable energy in cattle (Mori et al., 2025). Nevertheless, livestock production and productivity are limited due to seasonal changes that are faced with the seasonality of forage. Thus, the quantity and quality of feed sources affected animal productivity. Moreover, cattle could intake insufficient minerals such as calcium, phosphorus, and magnesium, which support cattle's health. Previous research elucidated that crossbreed cattle are less resilient to infectious tick-borne pathogens than native beef cattle (Kolte et al., 2017). Valls‐Fox et al. (2018) and Herrik et al. (2023) reported that free grazing leads cattle to ecological imbalances that may indirectly affect cattle health through altered disease dynamics and competition for resources. Overall, assessing the health status in free grazing systems involves multifaceted factors, influenced by behavioural patterns, nutritional quality of forage, environmental interactions, and management practices. Based on our knowledge, assessing health status in Khmer native x Hariana crossbred cattle-based in free grazing systems has not been extensively reported in previous studies. Consequently, the main objective of this study was to evaluate the health status of Khmer native × Hariana crossbred cattle under a free-grazing system, with particular emphasis on sex-based differences.
Materials and Methods
Animal, dietary treatment, and experimental design
The research was conducted at the Agricultural Research and Training Center (ARTC), National University of Battambang (NUBB), Chhras keo village, Kampong Preash commune, Sanke district, Battambang province, Cambodia (13°00'26.5"N; 103°18'49.0" E). The experimental cattle belonged to ARTC. The cattle housing was built under an iron, concrete floor, and zinc roof—the size of cattle housing (length 24 x width 8 x height 3) meters. At rest time (night and daytime), cattle were kept in the house to prevent hot temperatures and avoid any annoyance from harmful animals to cattle. In addition, surrounding housing cattle used mosquito nets and LED BULD (mosquito and insect repeller-20 watt). Clean water and rice straw were provided ad libitum. Ten heads of cattle were selected, consisting of five males (206.80 ± 10.15 kg/head) and five females (192.60 ± 12.59 kg/head) in terms of initial body weight (IBW). Those selected cattle were 18 months of age, with a 14-day feeding observation. The dietary treatments were assigned in a Completely Randomize Design with two treatments and five replications, each containing five cows. Feeding treatments were as follows: FGm, free-grazing (male), and FGf, free-grazing (female). Those cattle have natural grasses ad libitum in the experimental field in ARTC , from 8:00 am to 5:00 pm daily. The initial weigh, blood, and faecal sampling were collected in two rounds on 27 January 2024 and 10 February 2024 (first round and second round). This period falls within the dry season. Cambodia has two distinct seasons: the dry season from May to October and the rainy season from November to April. The outdoor temperatures recorded during the experiment indicate that the average temperature at 7:00 am was 24°C, at 12:00 pm was 33°C, and at 5:00 pm was 31°C, as shown in Figure 1.
Fig. 1. Outdoor temperature during the cattle experiment.
Experimental equipment
Faecal, blood collection, and analytical methods
The Ohaus PX224/E pioneer analytical balance, 220 g × 0.1 mg, external cal used for weighing cattle faecal and solid chemical. Erlenmeyer flasks 120 ml used for diluting faeces. The faeces were collected through the rectum of each cattle and kept in an ice box during transport to the laboratory, and those samples were kept in refrigerator ~ 40C. While eggs count, faecal is transferred from a freezer to an ice box and kept until faecal is thawed. This method is used to explore parasite eggs using the McMaster Counting: Modified McMaster Technique to identify the number 1 g of EPG. Procedure: Weigh 4 g of cattle faeces and place it in a beaker with a scale of 150 ml, add sodium chloride 60 ml, then stir it thoroughly with a fork or other appropriate material. Filter with a laboratory test sieve by size 250 μm and transfer to another sieve of size 53 μm to remove large pieces of grass debris. Using the pipette, withdraw the sample as the filtrate is being stirred to fill the compartment of the McMaster counting chamber slide, then take it to test under a compound microscope at 40 or 100 magnifications. To identify and count all eggs within the grid of both chambers were diluted at a ratio of (1:15 w/v) by mixing 2 g of fecal samples in a final volume of 30 ml. The McMaster chamber has a volume of 0.15 ml per grid; therefore, counting both grids represents a total volume of 0.30 ml. This volume corresponds to 1/50 of the diluted suspension, equivalent to 1 gram of feces (Zajac and Conboy, 2012). Parasite eggs in faecal 1 g are equal to the number of parasite eggs counted in both grids of McMaster chambers by multiplying the total of 50, which means that if we found “A” number of parasite eggs in 1 g, it is equal to A × 50 (Morm et al., 2021). Egg counts were used to grid each chamber of the McMaster, and eggs laid outside or over the grid were ignored. For instance, 10 parasite eggs counted in chamber I and 12 eggs in chamber II have to be calculated (10 + 12) × 50=1,100 EPG (Zajac and Conboy, 2012; Morm et al., 2021). A total of 10 ml of blood samples were collected from the jugular vein; 4 ml were kept in a test tube containing a sodium fluoride/Ethylenediaminetetraacetic Acid tube for glucose analysis, and each 4 ml was kept in a test tube containing a serum clot activator tube for blood urea nitrogen (BUN), creatinine, total protein, and triglyceride analysis and were sent to the LABORATOIRE HUMAN at road no.3, Battambang town, Cambodia. Blood urea nitrogen uses the urease method, creatinine uses the Isotope Dilution Mass Spectrometry traceable method, glucose (plasma) uses the glucose oxidase method, total protein (serum) uses the biuret lithium method, and triglyceride uses the glycerophosphate O method (Ruiz-Gutierrez et al. 1995; Küme et al. 2018; Zhang et al. 2022).
Growth performance
The average daily gain was determined using the IBW and final weight (FW). However, the IBW was weighted at the day-1 of experiment, and then at day-14 was weighted the FW. The body weight (BW) gain for the study was estimated from the IBW in Table 1, representative.
Table 1. Variable of an initial and final weight gain of cattle in the experiment.
Statistical analysis
The study was conducted using a CRD and analyzed using SPSS version 26. Independent T-tests were employed to compare treatment means. Differences were considered statistically significant at p < 0.05, while values of 0.05 <p < 0.10 were regarded as indicating a tendency. All results are reported as mean ± Std. Deviation. The statistical model was expressed as follows:
Yij=μ + Ti + Єij
where Yij=observed value of the dependent variable; μ=overall mean; Ti=fixed effect of the treatment; Єij=random error associated with the experimental unit under the treatment, assumed to be normally distributed with mean 0 and variance σ2. Pearson’s correlation coefficient (r) was used to evaluate relationships among weight gain, blood metabolites, parasite load, and microorganism characteristics.
Ethical approval
All operations were carried out in accordance with National University of Battambang (NUBB), under the guideline of Animal Care and Use for Scientific Purpose, with approval no. HEIP/2021 on 15 January 2021.
Results
A variable of an initial and final weight gain of cattle in the experiment
Table 1 shows that the initial average weight of cattle in the FGm group was 206.80 ± 10.15 kg, while the FGf group had an average of 192.60 ± 12.59 kg. The difference between the two groups at the beginning of the experiment was not statistically significant (p=0.085; p > 0.05). After 14 days, the final average weights were 210.90 ± 12.63 kg for FGm and 197.80 ± 12.45 kg for FGf, and no significant difference was observed between the groups at this stage as well (p=0.124; p > 0.05).
The first and second rounds of blood chemical characteristics
The blood chemical characteristics in Table 2 showed that glucose, triglycerides, creatinine, BUN, and total protein did not change (p > 0.05). Additionally, these blood chemical compositions were below the standards. For instance, glucose levels in the first and second rounds, lower than 74–110 mg/dl, affected powerlessness, hunger, and provoked animal stress. In this case, increased cortisol, adrenalin, and heart rate. However, animals may exhibit agitation and restlessness due to energy deficiency. Triglyceride levels are still insufficient, with a couple of readings lower than 150 mg/dl, which were collected over time, and varied with the animals' habitat. Furthermore, creatinine levels in the results were found to exceed the lab’s standard, leading the animals to serious kidney issues and powerlessness constraints. Moreover, BUN was very high, indicating that the kidneys were not functioning properly. Separately, the total protein is at a normal level.
Table 2. The blood chemical characteristics.
Body temperature, microorganisms, and parasite eggs
Body temperature in the first round (day 1) and the second round (day 14) did not change significantly (p > 0.05). However, Eimeria spp. and protozoa log cells/ml differed significantly between groups in the first round (p < 0.05). Eimeria spp. counts were higher in the FGm group than in the FGf group, whereas protozoa log cells/ml were lower in FGm compared with FGf. In contrast, neither Eimeria spp. nor protozoa levels differed between groups in the second round (p > 0.05), as presented in Table 3.
Table 3. Body temperature, microorganisms, and parasite eggs.
Pearson correlation (r) on weight gain, blood metabolites, parasites, and microorganism characteristics
Table 4 revealed that initial body weight (InW) exhibited a negative correlation with triglyceride levels during the first sampling period (r=−0.507, p=0.876) and a significant negative correlation with triglyceride levels in the second sampling period (r=−0.787, p=0.007). In contrast, the second body weight measurement showed no significant correlation with triglyceride levels (r=−0.834, p=0.003). Notably, Eimeria spp. demonstrated a very strong positive correlation with Proz1 (r=0.963, p < 0.001) and ambient temperature (r=0.907, p < 0.001) in the first sampling period. Similarly, during the second sampling period, Eimeria spp. maintained a strong positive correlation with Proz2 (r=0.965, p < 0.001).
Table 4. Pearson correlation (r) on weight gain, blood metabolites, parasites, and microorganism characteristics.
Discussion
Weigh gain of Khmer native x Hariana crossbreed cattle
The current studies presented in Table 1 found that the final weight gain of beef cattle was not positively affected by growth performance differences between male and female, as reported by Tofastrud et al. (2020), who noted that raising Charolais beef in free grazing with high-density stocking impacts animal performance. Generally, low-density grazing influences weight gain more than high stocking density in cows (Tofastrud et al., 2020). Previous studies indicated that increased use of sub-optimal habitats eventually leads to cows consuming less nutritious herbage in high stocking densities (Spedener et al., 2019), as predicted by ideal free distribution theory (Fretwell and Lucas, 1969). High stocking density affects weight gain in cows due to sward height and the availability of preferred feeding plants (Cornelissen and Vulink, 2015). Consequently, cows may still be unable to meet their nutritional requirements, even with sufficient grazing time when shorter swards are present (Chacon et al., 1978). According to Hessle et al. (2008) and Hansen et al. (2009), by comparing weight gain in native heifer breeds and Charolais, which is agreed upon in the current study. However, there is no effect on daily weight gain while grazing in the boreal forests of Norway among native heifer breeds (Braghieri et al., 2011; McCabe et al., 2019). However, this contrasts with findings by Dias et al. (2019), who argue that pasture-based dairy systems offer opportunities to improve herd nutrient use efficiency and milk production through strategic pasture allowance and supplementation strategies. The types of feed cattle consume change significantly when they graze in pastures. These variation changes are linked to feed intake and digestibility as well as losses in intestinal fill, which constitute a substantial portion of live weight (Spörndly et al., 2000; Hessle et al., 2007). The feeding regime before the grazing period directly impacts weight loss during the recovery phase, as measured by changes in live weight. So that it may ensure livestock regain weight efficiently after grazing. While pre-grazing might enhance the herd’s long-term health and productivity during effective management. A short grazing time is a main factor influencing weight loss, thereby leaving fewer days available for positively increasing weight gain before housing (Tofastrud et al., 2020).
The first and second rounds of blood chemical characteristics
The current studies found that serum glucose levels in both the first round and second round were lower than the healthy cows (74–110 mg/dl) standard, which is supported by Guzel et al. (2014) and Barson et al. (2019). There might be causes of hypoglycemia in Khmer native x crossbreed cattle. Which is increased peripheral glucose uptake, failure of gluconeogenesis or glycogenolysis, and endogenous hyperinsulinemia (Mukherjee et al., 2011). The serum glucose level is a crucial element in modulating reproduction, as lower levels affect the fertility rate (Yadav et al., 1995). In addition, the BUN in the first and second rounds was at a high level, which is related to fertility disorders; this symptom showed that the kidneys were not working well (Setiadi et al., 2003). The triglyceride observed, ranging from 20.40–26 mg/dl aligns with reported values for healthy cattle under normal feeding conditions (Smith et al., 2020), which influences dietary intake, energy balance, and metabolic demands. According to (National Research Council, 2016), triglycerides are higher in monogastric animals than in ruminants due to the microbes fermented in the rumen, which reduces dietary lipid absorption. Extremely low triglycerides (<20 mg/dl) may indicate energy deficits, poor nutrition, or metabolic disorders such as ketosis, though the current range does not suggest pathology. Prolonged undernutrition or high energy expenditure could suppress lipid mobilization (Herdt, 2000); the current range does not suggest pathology. However, prolonged undernutrition or high energy expenditure (e.g., lactation, growth) could suppress lipid mobilization (National Research Council, 2016). According to Kaneko et al. (2008), the observed range (1.5–1.7 mg/dl) slightly surpasses the usual reference intervals for adult cattle (0.8–1.4 mg/dl). Mild increases could be the result of subclinical renal impairment, temporary muscle catabolism, or dehydration. For instance, in grazing systems, heat stress or restricted water availability may temporarily raise creatinine (Radostits et al., 2007). However, persistently elevated levels warrant assessment of renal function via glomerular filtration rate and BUN to exclude chronic kidney disease or toxin exposure (Foster, 2023). Notably, native cattle breeds with higher muscle mass may exhibit elevated baseline creatinine without renal pathology, emphasizing the need for breed-specific reference ranges (Smith et al., 2020). In addition, crossbreeding of indigenous breeds is a common technique to improve productivity but set at risk local genetic resources (Tampaki et al., 2025).
Body temperature, microorganisms, and parasite eggs
Eimeria spp. oocyst counts in cattle ranged from 940 to 3,540 EPG in females and from 2,590–5,090 EPG in males. It was agreed that previous studies found above 500 EPG in ruminants (Najmuldeen et al., 2017). Daugschies and Joachim (2000) reported that high levels of intestinal parasites may be a factor in declining nutrient absorption, intestinal damage, and growth rate, particularly in young animals. The concurrent decline in protozoan populations (1.70–1.20 log cells/ml) may reflect dysbiosis in the gut microbiota, as protozoal communities are sensitive to shifts in pH and inflammation caused by Eimeria colonization (Williams, 1986). This inverse relationship between Eimeria load and protozoan density could indicate competition for resources or a destabilized microbial environment (Ellis et al., 2022).
Conclusion
Free grazing systems are an easy way to raise cattle in rural areas, as the animal’s owner does not need to have extensive land for feeding their animals. However, this method is not very beneficial to the animals' health. Animals consume insufficient nutrients or are contaminated by parasites or diseases. According to the current study, the free grazing system is very harmful to Khmer native x Hariana Crossbred cattle's health, leading to disease, parasite contamination, and altered blood chemical characteristics. To mitigate these challenges, livestock producers should prioritize providing balanced nutritional feed and implementing effective parasite control measures for their cattle. To be sufficient for Khmer native x Hariana crossbreed cattle, nutrient balance and anti-parasitics should be the main elements used for each animal. Furthermore, animal owners need to prepare a post-feeding at cattle housing to subsidise insufficient feed, while they are at free grazing during the daytime.
Acknowledgment
Authors are grateful to the project Innovate CAttle FArming System: Hydroponic Fodder for Sustainable Farming in North-West of Cambodia (ICAFAS), Higher Education Improvement Project of the National University of Battambang (NUBB), for supporting our research grant.
Conflict of interest
The authors declare that there are no conflicts of interest.
Funding
Authors are grateful to the Higher Education Improvement Project (HEIP) of the Ministry of Education, Youth, and Sport, Cambodia (Credit No. 6221-KH) for the financial support of this research.
Authors’ contributions
The authors contributed to the design of this study. Materials and data analysis were carried out by Sophany Morm. However, Mach Din, Vey Seb, and Kheng Kheak were carried out data collection. The first draft of the manuscript was written by Sophany Morm, Jinhu Huang, Jinxin Liu, Pao Srean, Phiny Chiv, Areerat Lunpha, and Ruangyote Pilajun. All the authors carefully checked and commented on the earlier manuscript version. The authors read and approved the final version of the manuscript.
Data availability
Data were not publicly available, but they are available from the corresponding author upon a reasonable request.
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