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Saudi Journal of Biological Sciences logoLink to Saudi Journal of Biological Sciences
. 2023 Aug 9;30(9):103768. doi: 10.1016/j.sjbs.2023.103768

Evaluation of trace elements in forages and their effect on gastrointestinal parasite burden in grazing sheep

Hafiz Muhammad Rizwan a,⁎, Muhammad Sohail Sajid b, Muhammad Younus c, Muhammad Ahsan Naeem d,e, Muhammad Sulman Ali Taseer c, Hossam Ebaid f
PMCID: PMC10461023  PMID: 37645686

Abstract

This study was designed to evaluate the trace elements (minerals) in forages fed to sheep and their effect on gastrointestinal parasite burdens. The ultimate objective was to determine the correlation between the burden of gastrointestinal (GI) parasites and the level of trace minerals in sheep serum as a result of the forages they grazed on. A total of 384 faecal samples were collected from sheep in each of the districts (Sialkot and Multan) and examined quantitatively using the McMaster technique. Serum collected from them and plants were pre-treated, and spectrophotometry was used to determine the concentration of trace minerals (Mn, Co, Cu, and Zn). The level of these trace elements differed significantly (P < 0.05) in forages from both districts. In the district of Sialkot, the highest concentrations (mg/Kg) of Zn (38.53 ± 0.16) were found in Cichorium intybus, Cu (41.57 ± 0.07) in Cynodon dactylon, Mn (39.61 ± 0.05) in Parthenium hysterophorus, and Co (1.42 ± 0.03) in Coronopus didymus. In the district of Multan, the highest concentrations (mg/Kg) of Zn (39.43 ± 0.46) were found in Cichorium intybus, Cu (25.76 ± 0.36) in Cynodon dactylon, Mn (34.29 ± 0.53) in Launaea nudicaulis, and Co (1.74 ± 0.08) in Brachiaria raptens. The prevalence of GI parasites in sheep populations in district Sialkot was 34%, while in district Multan, it was 32%. In tehsil Sialkot of district Sialkot, Zn and Cu were significantly (P < 0.05) correlated with eggs per gram (EPG) of faeces, while in tehsil Multan City of district Multan, only Cu was significantly (P < 0.05) correlated with EPG. The potential mechanism behind the role of trace minerals in lowering the burdens of GI parasites requires more investigation. It is recommended that plants with high content of trace minerals should be utilized as part of comprehensive preventive and control strategies against GI parasitism in ruminant animals like sheep.

Keywords: Trace minerals, Gastrointestinal parasites, Sheep, Forages, Pakistan

1. Introduction

A global challenge in the livestock industry is to maintain and improve gut health and immunity to obtain optimal production. To improve and maintain gut health, it is important to assess the associated risk factors, such as parasite burden, forage quantity and nutrient profile of forages (Fekete and Kellems 2007). A diet with mineral supplementation is essential to optimize metabolic processes in animals, as trace minerals act as cofactors for various important enzymatic reactions (Erdoğan et al. 2002). Grazing sheep are popular and provide a comparatively cheap source of red meat source in Islamic countries like Pakistan, not only for routine consumption, but also for religious events like Eid ul Azha (Montossi 2013).

The amount of trace minerals required for normal metabolism in animals is very small, usually < 100 mg/Kg (McDowell, 1992), and their quantity in the serum of animals is also very low, i.e., around 2 ug/mL (Suttle, 2010). Trace minerals play significant roles in immunity and gut health, even though they are required in very low amounts. Trace minerals contribute to growth, production, and reproduction (Rizwan et al., 2021). For fully functional immunity, certain enzymes are required, and trace minerals are cofactors for these enzymes. When an animal is deficient in trace minerals, it becomes more susceptible to certain diseases as its ability to fight against them is reduced (Arthington and Ranches, 2021).

Understanding of the importance of trace minerals for better performance of animals is increasing day by day, and as a result, researchers have suggested only a few changes in minimal requirements of trace minerals over the last decades (Greene, 2000). The quantity of trace minerals in grazing animals is determined mainly by the mineral profile of the forages eaten, while the mineral profile is affected by several factors, including forage species, fertilization, maturity, and pH of the soil. Animals can store trace minerals in their bodies, which they can use when a short-term deficiency of them is encountered (McDowell and Arthington, 2005). Supplemental nutrients are given to livestock animals in developed countries to help counter deficiencies of energy and protein in forages. The bioavailability of trace minerals can be compromised by antagonists present in food supplements and forages (Spears and Weiss, 2014).

Almost every country in the world faces specific implications related to trace mineral supply that affect management systems (Arthington and Ranches, 2021). Pakistan is gifted with a variety of ecological, climatic, and topographical zones. Animal production is largely based on natural vegetation and agro-grazing in Pakistan. Local conditions and climate not only influence the type and nature of plants, their composition, distribution, and nutritional value, but also affect the types and numbers of grazing animals. Therefore, different grazing sites and cropping belts in Punjab, Pakistan, have different effects on the chemical and botanical composition of grazing forages. Surveys carried out in various regions of Punjab show that many plant species are used by small ruminants for grazing (Ullah et al., 2012, Abdullah et al., 2013, Qudoos et al., 2017).

Many plant species have the potential to reduce parasite loads in sheep (Marley et al., 2003), but the exact mechanism of action is unknown (Suttle et al., 1992). In this study, we hypothesized that the trace element profile of sheep may be linked with parasite immunity and may decrease faecal egg count (FEC) in sheep under natural conditions. The present study was planned to carry out the following objectives: quantitative and qualitative estimation of gastrointestinal (GI) parasites in sheep in targeted study areas; calculation of concentrations of trace minerals (Zn, Mn, Cu, and Co) in the native breeds of sheep and fodder; and determination of correlation between parasite burden and level of trace elements in sheep in study areas.

2. Materials and methods

2.1. Study areas

The study was conducted in two districts, including district Sialkot (32° 30′0″ N / 74° 31′0″ E), consisting of three tehsils (a) Sialkot, (b) Daska, and (c) Pasroor; and Multan district (30° 11′44″ N / 71° 28′31″ E), which has four tehsils (a) Multan Saddar, (b) Multan City, (c) Jalalpur Pirwala, and (d) Shujabad.

2.2. Coprological examination

The study population consisted of native sheep breeds from the districts of Multan and Sialkot. Using standard protocols, 384 faecal samples were collected from sheep in each district. To assess the endo-parasitic load i.e., egg per gram (EPG), the Modified McMaster test was used, and a quantitative faecal examination was carried out as described by Soulsby (1982). Standard identification keys were used to identify parasite eggs.

2.3. Forage sampling and pre-treatment

Forages commonly grazed by animals were collected in triplicate from study area grazing sites. The samples were collected in polythene bags and labelled for shipment to the University of Agriculture, Faisalabad, Pakistan. Samples were identified by a professional from the Botany Department, University of Agriculture, Faisalabad. The forage leaves were washed with HCL (1%) followed by distilled water then dried first in the open air and after that placed in a drying oven (65 ± 5°C). An electrical grinding machine was used to grind dried leaves, which were then subjected to wet digestion (Miller, 1998). In short, one g dried sample of the plant was digested with HClO4 (perchloric acid) and HNO3 (nitric acid) in a 1:3 ratio on a hot plate. After clearing the material, the final volume was maintained at 50 mL with de-ionized water, and then filtration was performed through Whatman filter paper No. 42 and the filtrate was kept in bottles with no airflow for mineral profile determination.

2.4. Sera sampling and pre-treatment

A total of 384 blood samples of sheep from each district were collected using a standard protocol (Soulsby, 1982). After centrifugation, serum was separated and used for wet digestion using a method provided by AOAC (2005). A digestion flask was first filled with one mL serum and 10 mL HNO3 (in concentrated form) and heated at 60–70°C for 15 min. Five mL HClO4 was added to the flask after cooling. The flask contents were heated until only 2 mL of the mixture remained. After that, deionized water was added to the mixture to make the total volume 25 mL.

2.5. Mineral analyses

An atomic absorption spectrophotometer (Hitachi Polarized Zeeman AAS, Z-8200) was used to measure the concentration of Zn, Cu, Co, and Mn using standard protocols provided by Anan et al. (2001). Standard solutions for Zn, Cu, Mn, and Co were prepared using reagent-grade salts. Each standard solution was run individually for these minerals and their absorbance was recorded. Standard curves for each mineral were constructed by plotting the absorbance of standards against their concentrations. The concentrations of the respective minerals in the samples were calculated from their standard curves.

2.6. Statistical analyses

To identify variations in the trace mineral profiles of sera and forages, data were subjected to analysis of variance (ANOVA). Pearson's correlation method was used to determine the relationship between serum trace mineral concentrations and the burden of GI parasites. Statistically, P-values of < 0.05 were regarded as significant (Schork and Remington, 2010). The Minitab 17 software was used for all statistical tests.

3. Results

A total of nine species of forages were collected from district Sialkot and 10 from district Multan (Table 1). During the survey, five species of forages (Digitaria sanguinalis, Cichorium intybus, Cynodon dactylon, Medicago polymorpha, and Trifolium alexandrinum) were common in both districts. The level of selected trace minerals varied significantly (P < 0.05) in different forages in both districts. In district Sialkot, the highest concentrations (mg/Kg) of Zn (38.53 ± 0.16) were found in Cichorium (Ci.) intybus, Cu (41.57 ± 0.07) in Cynodon (C.) dactylon, Mn (39.61 ± 0.05) in Parthenium hysterophorus, and Co (1.42 ± 0.03) in Coronopus didymus. In district Multan, the highest concentrations (mg/Kg) of Zn (39.43 ± 0.46) were found in Ci. intybus, Cu (25.76 ± 0.36) in C. dactylon, Mn (34.29 ± 0.53) in Launaea nudicaulis, and Co (1.74 ± 0.08) in Brachiaria raptens. The mean level of Mn, Co, Cu, and Zn in forages species collected from these districts are shown in Table 1. Among different tehsils, mean concentrations of these trace minerals in forages did not differ significantly (P > 0.05; Table 2). Mean concentrations of Zn and Cu in the serum of sheep grazed in different tehsils of district Sialkot varied significantly (P < 0.05), while Mn and Co showed non-significant variation (P > 0.05). In district Multan, mean concentrations of all the selected trace elements in the serum of sheep grazing in different tehsils varied significantly (P < 0.05; Table 3).

Table 1.

Mean ± SE concentrations (mg/Kg) of specific trace minerals (Mn, Co, Cu, and Zn) in forage collected from Sialkot and Multan districts in Punjab, Pakistan.

Districts Forages Mn Cu Zn Co
Sialkot Cichorium intybus 15.45 ± 0.22de 34.48 ± 0.11b 38.53 ± 0.16a 1.11 ± 0.05a
Trifolium alexandrinum 19.54 ± 0.04 cd 28.50 ± 0.07c 35.21 ± 0.07a 1.07 ± 0.02a
Mazus reptans 18.59 ± 0.04de 24.50 ± 0.13d 35.20 ± 0.11a 1.23 ± 0.05a
Medicago polymorpha 23.52 ± 0.12c 25.14 ± 0.09 cd 34.79 ± 0.07a 1.09 ± 0.05a
Digitaria sanguinalis 37.16 ± 0.07b 39.14 ± 0.09b 30.37 ± 0.11b 1.08 ± 0.05a
Malva neglecta 31.48 ± 0.04b 26.18 ± 0.06 cd 28.60 ± 0.16b 1.29 ± 0.06a
Cynodon dactylon 14.83 ± 0.06e 41.57 ± 0.07a 26.17 ± 0.13b 0.89 ± 0.03a
Parthenium hysterophorus 39.61 ± 0.05a 40.72 ± 0.05a 24.53 ± 0.06c 1.04 ± 0.03a
Coronopus didymus 29.07 ± 0.14b 23.17 ± 0.19d 18.44 ± 0.16c 1.42 ± 0.03a
Multan Cynodon dactylon 31.58 ± 0.20c 25.76 ± 0.36a 39.41 ± 0.27a 1.34 ± 0.07abc
Cichorium intybus 27.87 ± 0.83d 24.20 ± 0.57a 39.43 ± 0.46a 1.53 ± 0.06ab
Convolvulus arvensis 24.41 ± 0.17ef 12.45 ± 0.15e 36.71 ± 0.39abc 0.81 ± 0.09d
Brachiaria raptens 18.36 ± 0.18 h 19.57 ± 0.17b 35.44 ± 0.24abcd 1.74 ± 0.08a
Euphorbia prostrata 22.64 ± 0.19 fg 13.33 ± 0.42d 33.14 ± 0.30abcd 1.04 ± 0.04 cd
Trifolium alexandrinum 21.29 ± 0.33 g 13.46 ± 0.18d 32.54 ± 0.43abcd 1.64 ± 0.11a
Medicago polymorpha 33.71 ± 0.48bc 19.27 ± 0.63b 32.24 ± 0.60abcd 1.01 ± 0.08 cd
Ziziphus mauritiana 26.95 ± 0.57d 21.45 ± 0.65ab 29.75 ± 2.44bcd 0.95 ± 0.31d
Digitaria sanguinalis 33.64 ± 0.28abc 18.72 ± 0.57bc 28.35 ± 0.31bcd 1.34 ± 0.09abc
Launaea nudicaulis 34.29 ± 0.53ab 16.36 ± 0.11c 25.23 ± 0.24d 1.72 ± 0.09a
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In a column, the mean sharing similar letters is not statistically significant (P > 0.05).

Table 2.

Mean ± SE concentrations (mg/Kg) of specific trace minerals (Mn, Co, Cu, and Zn) in forage collected from different administrative divisions (tehsils) of Sialkot and Multan districts in Punjab, Pakistan.

Districts Tehsils Mn Cu Zn Co
Sialkot Pasroor 27.12 ± 6.70a 29.37 ± 2.03a 33.31 ± 3.99a 0.93 ± 0.30a
Sialkot 23.16 ± 5.08a 26.05 ± 3.74a 30.40 ± 7.33a 1.11 ± 0.23a
Daska 25.57 ± 9.12a 27.70 ± 4.94a 27.85 ± 4.74a 0.94 ± 0.08a
Multan Multan City 28.54 ± 4.40a 19.42 ± 4.48a 32.13 ± 5.30a 1.34 ± 0.38a
Multan Sadar 26.56 ± 5.28a 21.84 ± 3.14a 30.64 ± 1.26a 1.52 ± 0.16a
Shujabad 27.75 ± 6.54a 19.37 ± 3.74a 34.42 ± 3.45a 1.59 ± 0.26a
Jalal Pur Pirwala 28.33 ± 3.62a 18.14 ± 5.86a 35.13 ± 2.52a 1.23 ± 0.37a
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In a column, the mean sharing similar letters is not statistically significant (P > 0.05).

Table 3.

Mean ± SE concentrations (mg/L) of specific trace minerals (Mn, Co, Cu, and Zn) in serum of sheep collected from different administrative divisions (tehsils) of Sialkot and Multan districts in Punjab, Pakistan.

Districts Tehsils Mn Cu Zn Co
Sialkot Pasroor 0.22 ± 0.05a 0.74 ± 0.08c 0.90 ± 0.28b 0.27 ± 0.33a
Sialkot 0.25 ± 0.11a 1.09 ± 0.09a 1.27 ± 0.21a 0.29 ± 0.38a
Daska 0.21 ± 0.11a 0.96 ± 0.17b 0.76 ± 0.08b 0.26 ± 0.55a
Multan Multan City 0.13 ± 0.05ab 0.85 ± 0.16a 0.87 ± 0.17a 0.21 ± 0.04a
Multan Sadar 0.24 ± 0.07a 0.59 ± 0.06b 0.90 ± 0.14a 0.17 ± 0.03b
Shujabad 0.18 ± 0.03b 0.54 ± 0.07b 0.89 ± 0.08a 0.12 ± 0.02c
Jalal Pur Pirwala 0.17 ± 0.06b 0.79 ± 0.08a 0.72 ± 0.13b 0.11 ± 0.02c
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In a column, the mean sharing similar letters is not statistically significant (P > 0.05).

The concentration of trace elements in individual plants varied significantly in both districts. The mean concentration of Mn in individual forages was lower than the critical value (40 mg/Kg). However, the concentration of Cu and Co was higher than the critical values (10 mg/Kg and 0.125 mg/kg, respectively) in each forage of the study districts. Four forage species from district Sialkot and three species from district Multan showed a lower level of Zn concentration than the critical value (30 mg/Kg) required for sheep.

The concentration of trace elements in individual plants varied significantly in both districts. The overall mean concentration of Mn in forages of different tehsils of the study districts was lower than the critical value (40 mg/Kg). However, the concentration of Cu and Co was higher than the critical values (10 mg/Kg and 0.125 mg/kg, respectively) in forages collected from different tehsils of the study districts. The overall mean concentration of Zn in forages collected from different tehsils of the study district was in the range of or more than the critical value (30 mg/Kg), except for one tehsil in Sialkot district.

Analysis of blood is an accepted method for determining the levels of trace minerals in animals. About < 2 mg/L of trace minerals is maintained in animal serum. Mean concentrations trace elements in serum of sheep collected from different tehsils of study districts was in rage.

The prevalence of GI parasites in sheep populations of district Sialkot was 34%, while in district Multan, it was 32%. In district Sialkot, the burden of GI parasites was highest in tehsil Pasror, while tehsil Sialkot had the lowest burden (P < 0.05). In district Multan, tehsil Shujabad showed the highest burden, followed in order by tehsil Jalal Pur Pirwala, Multan Sadar, and Multan City (Table 4). Pearson’s correlation of the concentration of trace minerals in serum with the mean EPG values in different tehsils of the study districts (Sialkot and Multan) of Punjab, Pakistan, is given in Table 5. In tehsil Sialkot of district Sialkot, Zn and Cu were significantly (P < 0.05) correlated with EPG, while in tehsil Multan City of district Multan, only Cu was significantly (P < 0.05) correlated with EPG.

Table 4.

Burden of gastrointestinal parasitic infection of sheep in different tehsils of study districts (Sialkot and Multan) of Punjab, Pakistan.

Districts Tehsils Light (%) Moderate (%) High (%) χ2 P-value
Sialkot Pasror 22 (19.13) 24 (20.87) 17 (14.78) 0.852 0.021
Sialkot 7 (5.65) 6 (4.84) 6 (4.84)
Daska 18 (12.41) 15 (10.34) 16 (11.03)
Multan Multan City 4 (4.04) 6 (6.06) 4 (4.04) 1.001 0.986
Multan Sadar 9 (11.25) 11 (13.75) 8 (10.00)
Shujabad 14 (12.96) 16 (14.81) 13 (12.04)
Jalal Pur Pirwala 15 (15.46) 12 (12.37) 11 (11.34)
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Light = 100–––600; Moderate = 601–1000; High = > 1000.

Table 5.

Correlation of the specific trace minerals in serum with mean parasitic eggs per gram of faeces in different tehsils of study districts (Sialkot and Multan) of Punjab, Pakistan.

District Tehsils Mn Cu Zn Co
Sialkot Pasror −0.028* −0.007* −0.056* −0.150*
0.746** 0.933** 0.520** 0.085**
Sialkot −0.113* −0.157* 0.155* 0.057*
0.210** 0.014** 0.017** O.565**
Daska 0.014* −0.014* −0.054* 0.018*
0.864** 0.862** 0.512** 0.828**
Multan Multan City −0.110* 0.059* −0.128* −0.027*
0.316** 0.020** 0.243** 0.805**
Multan Sadar 0.091* −0.117* −0.048* 0.075*
0.418** 0.300** 0.668** 0.508**
Shujabad 0.042* −0.030* −0.064* −0.031*
0.649** 0.749** 0.488** 0.741**
Jalal Pur Pirwala −0.164* 0.073* −0.035* 0.020*
0.102** 0.470** 0.733** 0.846**
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*= Pearson Correlation, **= Sig. (2-tailed), Bold = Significant correlation.

Pearson’s correlation demonstrated significant (P < 0.05) correlation of EPG with Zn and Cu in tehsil Sialkot of district Sialkot, and with Cu in tehsil Multan City of district Multan. All other tehsils of study districts showed non-significant (p > 0.05) correlation of EPG with trace element profiles of sheep reared on the rangelands of different tehsils.

4. Discussion

The rangelands of an area are crucial to its economy since they are utilized for grazing animals. Pakistan's topographical areas, climatic conditions, and ecological zones, are ideal for supporting a wide range of edible natural flora. Grazing is used to raise over 60% of Pakistan's livestock populations. The forage species found in this study and those by Ahmad et al., 2011, Mashwani et al., 2012, and Qudoos et al. (2017) are similar in some cases. During their surveys, other authors also discovered different species of forages compared to our study (Hussain and Durrani, 2008, Ahmad et al., 2011). Many types of forages were identified by researchers in Kenya, Norway, and Botswana which are consumed by their animal populations (Aganga and Mesho, 2008, Lengarite et al., 2013, Nordlokken et al., 2015). The geography of an area, the composition and structure of soil, and meteorological conditions contribute to differences in plant diversity (Liu et al., 2020). Also, some data suggest that grazing can affect the variety of forages (Bainard et al., 2020).

Edible forages are a significant source of trace minerals for livestock in a natural grazing ecosystem. In the present study, nine edible species of forages were collected from district Sialkot and 10 from district Multan; among these, five species were common in both districts. The variation in the presence of different edible forage species in the study districts might be due to variations in the ecosystem, egro-climate, environment, and topography of the area. Since the trace mineral composition of edible forages represent the level of minerals in grazing livestock, evaluation of trace minerals in forages should be a regular practice (Hoste et al., 2006, Faulkner and Weiss, 2017). Similar to our study, Qudoos et al. (2023) found significant variation in the concentration of trace elements in the collected forages. According to Jank et al. (2014), the mineral contents of forages are influenced by the soil type, age and species of plant, use of fertilizer and season.

Analysis of blood is an accepted method for determining the levels of trace minerals in animals (Spears et al., 2022). Only around 100 mg/kg of dry matter of trace minerals are needed overall for animals (NRC, 2016). However, < 2 mg/L of trace minerals is maintained in animal serum (Suttle, 2010). In the present study, the mean concentrations of trace elements in the serum of sheep collected from different tehsils of the study districts were in range, i.e., <2 mg/L. In another study conducted by Erdoğan et al. (2002), the level of Se, Ca, Na and Mg was within the normal range in both goat and sheep. However, Cu and Zn levels were below essential levels in every region, although K levels were somewhat lower in some. Sex, age, breed, production capacity, and genotype are variables that might affect the quantity of and requirements for trace elements in serum (Devi et al., 2011, López-Alonso, 2012, Yatoo et al., 2013). The level of trace minerals in animals, notably Zn, are affected by several variables, including sex, age, health, stress, and nutrition (Ishag et al., 2014, Abdelrahman et al., 2022). Because animals grow quickly and their food may contain inhibitors, variations in trace mineral concentration may also be observed (López-Alonso, 2012).

Epidemiology is the foundation for the control of parasitic infections. In comparison to other districts of Pakistan, the prevalence of GI parasites was lower in this study (Shah et al., 2015, Batool et al., 2022, Ruhoollah et al., 2023). Moreover, GI parasite prevalence found in many places of the world is higher than reported here (Singh et al., 2017, Bhowmik et al., 2020). Similar to our investigation, Kantzoura et al. (2012) in Greece and Latera et al. (2016) in Ethiopia found lower GI parasite frequencies. Numerous variables, including grazing practices, education level, standard of management, irrational use of anthelmintics, seasonal differences, variations in the agro-ecology of the study areas, and financial capacity of the farmers can affect parasite prevalence. In addition, farms lacking fences, having poorly draining soil, the grazing of female and male animals and adult and young animals together, the predominant agro-climatic conditions, and overcrowding of animals are other factors that affect the intensity of parasitism (Lashari and Tasawar, 2011, Nana, 2016, Jansen et al., 2020).

EPG is a useful and efficient monitoring method for determining GI parasite load in animals (Poulin, 2007). EPG values in the present study are consistent with those reported by others (Al-Shaibani et al., 2008, Roy et al., 2013). However, there is also evidence of both higher (Qudoos et al., 2017) and lower (Martínez-Valladares et al., 2013) values of EPG. The sampling size, inhibition of egg production, exposure of faeces to the environment, and moisture contents are possible causes of variation in EPG values (Bricarello et al., 2004, Das et al., 2011). Seasons, sex, and age are additional variables that could affect EPG (Idika et al., 2012, Roy et al., 2013). The low level of EPG we found may be attributed in part to the availability of a sufficient amount of nutrient-rich pasture. Nutrient-rich pasture can boost animal immune systems, reduce the growth and reproductive potential of parasites, and ultimately lower EPG (Kumar et al., 2013, Pathak, 2017). Moderate to high EPG may result if animals acquire a high infection rate during the rainy season (Githigia et al., 2005).

Trace mineral deficiencies pose a major risk to grazing animals. Animals primarily need trace minerals for healthy immune systems and resistance to disease (Radostits et al., 2007, Weyh et al., 2022). Sheep flocks with ideal serum Cu and Zn levels generally have lower EPG. In the present study, the sheep population of Pasroor and Daska tehsils of Sialkot district showed a high intensity of parasitic infection. Due to this high intensity of parasitic infection, the level of trace elements in the sheep serum of these tehsils was lower. However, the intensity of parasitic infection in the sheep population of tehsil Sialkot was lower, and the level of trace elements in the sheep serum of this tehsil was higher. Similar to our study Hill and Shannon (2019) concluded that animals with parasitic infections or other disorders have lower serum Zn and Cu levels than uninfected animals. Schafer et al. (2015) observed a non-significant finding relating the low EPG in comparison to the previously indicated combination. Animals given Zn supplementation experienced a low level of parasitic infections (Fançony et al., 2022), and animals with Zn deficiency experienced a high level of worm burden (Qudoos et al., 2017). In the current investigation, an insignificant correlation between EPG and serum Mn and Co was found. Low levels of Co make animals more prone to acquiring parasites; for instance, higher EPG levels were seen in animals fed diets containing low amounts of Co (Vellema et al., 1996, González-Montaña et al., 2020).

5. Conclusions

Five species of forages were common in the selected study areas, which had different agro-climatic conditions. The concentration of trace minerals in forages was significantly different in both districts. The level of Mn in the forages of both districts was lower than the critical values required for animals, while the level of Cu was higher and the level of Zn was higher/normal in many forages and marginal in some species. It has been noted that the majority of the sheep populations in various tehsils of selected areas had normal or higher levels of trace minerals. Trace minerals like Zn and Cu play a significant role in lowering GI parasite burdens in naturally-infected sheep. There is a need to understand how different trace minerals protect against parasites and reduce parasites in animals.

Funding

Financial support for the project was provided by the Higher Education Commission (HEC), Islamabad under the project No. 20–2666/NRPU/R&D/HEC/12/6974, titled “Phytotherapy: An Easy and Economic Way to Cure the Gastro-intestinal Parasites in Sheep” and Researchers Supporting Project number (RSP2023R366), King Saud University, Riyadh, Saudi Arabia.

7. Institutional review board Statement

This study was approved by the Research Ethics Committee, Faculty of Veterinary Science, University of Agriculture, Faisalabad, Pakistan. The standard guidelines for institutional animal care and use (IACU), University of Agriculture, Faisalabad, Pakistan, were followed.

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgments

Authors are very thankful to Emeritus Professor Dr. Timothy G. Geary, McGill University, McDonald Campus, Quebec, Canada, for English language revision of the manuscript. The authors extend their sincere gratitude to the Higher Education Commission, Islamabad, Pakistan, and King Saud University, Riyadh, Saudi Arabia for generously funding the completion of this research and covering the publication charges through project No. 20–2666/NRPU/R&D/HEC/12/6974 and Researchers Supporting Project Number RSP2023R366, respectively.

Footnotes

Peer review under responsibility of King Saud University.

Contributor Information

Hafiz Muhammad Rizwan, Email: hm.rizwan@uvas.edu.pk.

Muhammad Sohail Sajid, Email: drsohailuaf@uaf.edu.pk.

Muhammad Younus, Email: younusrana@uvas.edu.pk.

Muhammad Ahsan Naeem, Email: ahsan.naeem@uvas.edu.pk.

Muhammad Sulman Ali Taseer, Email: sulman.ali@uvas.edu.pk.

Hossam Ebaid, Email: habdrabou@ksu.edu.sa.

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Further Reading

  1. Pecora F., Persico F., Argentiero A., Neglia C., Esposito S. The role of micronutrients in support of the immune response against viral infections. Nutrients. 2020;12(10):3198. doi: 10.3390/nu12103198. [DOI] [PMC free article] [PubMed] [Google Scholar]
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  3. Waller P.J., Bernes G., Rudby-Martin L., Ljungstrom B.L., Rydzik A. Evaluation of copper supplementation to control Haemonchus contortus infections of sheep in Sweden. Acta Veterinaria Scandinavica. 2004;45:149–160. doi: 10.1186/1751-0147-45-149. [DOI] [PMC free article] [PubMed] [Google Scholar]

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