Blood content of selenium and zinc of female alpacas and their offspring raised in the Andes.

doi:10.21203/rs.3.rs-2953793/v1

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Blood content of selenium and zinc of female alpacas and their offspring raised in the Andes.

https://doi.org/10.21203/rs.3.rs-2953793/v1

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Minerals are important for animals in many biological functions, there is scarce information however about micromineral content in blood of South American camelids under their prevalent production system. This study aims to determine the content of selenium in whole blood and zinc in blood serum of grazing alpacas at three different physiological states in the Peruvian Andes. Fifteen female alpacas on native pastures of the Andes were included in this study. Blood samples were collected for measurement of selenium in whole blood and zinc in blood serum from female alpacas at pre-calving, ten days after calving, and late lactation, as well as their respective offspring. Selenium and zinc content of pastures consumed by alpacas also were determined. The content of Se and Zn of the diet selected by alpacas met the requirements for the physiological states they go through in comparison with literature references. The concentration of zinc in blood serum of female alpacas was higher ten days after calving (0.26 µg/ml), compared to pre-calving and late lactation. The same tendency was found in the case of their offspring at ten days after calving (0.23 µg/ml) higher than for late lactation. Unlike zinc, in the case of selenium, no differences were found between the physiological states studied for alpacas or their offspring. Apparently, the zinc serum concentration changes through productive cycle of alpacas but not the whole blood selenium concentration. Further investigation is needed to define alpaca adequacy of zinc and selenium in relation to blood content of both minerals.

Alpaca

Selenium

Zinc

Blood

Andes

Nutrition

Minerals fulfill essential functions for life; being fundamental in the productive, reproductive, and immunological performance of livestock animals. Selenium is related to antioxidant activity and animal growth, being part of the enzyme glutathione peroxidase and the enzyme 5-monodeiodinase (McDowell 2003a). Zinc participates among other functions as a cofactor and integral component of molecules (structural or functional), having immunological, metabolic and performance repercussions (McDowell 2003b). The status of a mineral is defined as the determination of the concentration of mineral in the soil, diet, and blood level and the interpretation of the results to obtain a panoramic view of the impact of the mineral on the productive performance of the animal. However, in some cases, the analysis of minerals is only carried out in blood samples, since the samples of the diets that the animals consume are difficult to determine due to the amount ingested and their composition, as well as the different factors that could alter the concentration of a certain mineral (source, composition, and interaction) (Herdt and Hoff 2011).

There is scarce information regarding the mineral content of blood and the effects of physiological states on it, in the alpacas reared in the Andes, with almost the majority of information coming from Europe (Husakova et al. 2014; Pechová et al. 2018), North America (Van Saun and Herdt 2014), and Oceania (Judson 1999; Judson et al. 2011) all with important differences in the environment and diet to the prevalent situation in the Andes. Specifically, the concentrations of selenium and zinc in the pastures consumed by grazing alpacas in the Andes are unknown, as are their concentration in the blood; not knowing whether both microminerals are at adequate or deficient levels and the impact of the different physiological states on them. Therefore, the objective of our work was to determine the content of selenium in whole blood and zinc in blood serum in grazing alpacas (mother and offsprings) in the Peruvian Andes at critical physiological states (gestation and lactation).

Location

This study was conducted in the district of Ocuviri,department of Puno, at a height of 4397 meters above sea level. The temperature during the year had a minimum of -11.2°C and a maximum of 21.8°C, with annual precipitation of 843 mm (SENAMHI 2020). The alpacas were in extensive production, being fed based on the native pastures from the area without any kind of supplementation.

Animals

We worked with three physiological states for female alpacas and their offsprings, which were: pre-calving, ten days after calving, and late lactation.

It was projected to work with 30 Suri breed female alpacas, which completed the three physiological states, however, due to deaths of the offspring and abortions of the female alpacas, only 15 alpacas were counted at the end of the study, which went through the three physiological states. These female alpacas at the beginning of the evaluation were at three months before calving with a body condition score of 2.86 ± 0.41 and an estimated age of 4.03 ± 0.93 years. Body condition was evaluated as suggested by Van Saun (2009), and the estimated age through dental identification as suggested by Aragon (2005).

Sampling and analytical procedure

Blood samples were taken at three different moments: in the beginning of the rainy season (November 2018), alpacas in prepartum, with 37 ± 29.9 days before calving; in the middle of the rainy season (February 2019), female alpacas and their offspring, after 10 days of calving and being born, respectively (postpartum); and in the dry season (July 2019), female alpacas and their offspring, both in late lactation, with 162 ± 7.7 days after calving.

The technique used for blood sampling was external jugular vein puncture, extracting 5 ml of blood from both mothers and offspring. Blood samples were collected in tubes with anticoagulant (EDTA) and without anticoagulant, for selenium and zinc respectively. For the analysis of zinc, the formation of the clot was expected in the collected tubes. To separate the serum from the clot formed, the samples were centrifuged for 10 minutes at 2500 rpm, extracting the supernatant (serum) and placing it in Eppendorf tubes, keeping them at -20°C. A similar activity was carried out with the tubes that had the blood with anticoagulant, omitting the previous step of centrifugation. 15 samples were collected for each physiological state, where the female alpacas had three physiological states and the offspring had two physiological states.

Pasture samples from diets that consume the female alpacas were collected at the same time (month and year) as described above for the blood samples. A manual simulation technique was used, which consists of observations of the individual grazing of the animals for two to three hours, near the grazing area, to later take representative manual samples of the natural grass consumed by the animal (Austin et al. 1983). These samples were collected from the observation of two alpacas from the herd, each with 30 feeding stations, cutting the forage at an approximate height of 5 cm above the ground (Burton et al. 2003), averaging these 30 food stations as a single sample, having at the end two samples from the two alpacas. In the case of zinc, the two samples obtained were analyzed, however, in the case of selenium, a single sample from the mixture of the two samples taken was analyzed.

Selenium in whole blood and pasture was measured by Unidad de Servicios de Análisis Químicos de la Facultad de Química e Ingeniería Química (UNMSM) using a graphite furnace atomic absorption spectrophotometry equipment (SHIMADZU, model AA6800). Zinc in serum blood and pasture was measured according to Soltero et al. (2007), and Bazán (2018) respectively, in the Laboratorio de Evaluación Nutricional para Animales, Facultad de Zootecnia (UNALM) using a flame atomic absorption spectrophotometry analysis (GBC model SavantAA ∑).

Statistical analysis

For analysis of data from whole blood selenium and blood serum zinc, fixed block designs were carried out considering the alpacas as block and the physiological states as a factor (independent variable), two separate analyzes were made, one for female alpacas and other for their offspring. For female alpacas we considered three levels for physiological factor (pre-calving, ten days after calving, and late lactation) and fifteen fixed blocks (mother alpacas), and for their offspring two levels for physiological factor (ten days after calving and late lactation) and fifteen fixed blocks (offspring alpacas). In the case to found differences between the physiological states a Least significant difference test (LSD) was made. All the analysis designs were run by Statistical Analysis System (SAS OnDemand for Academics).

Zn and Se content in pastures

No differences were observed among physiological states in the concentration of selenium in the pastures that consumed the female alpacas, the value was 0.28 ± 0.04 mg/Kg. The same was for zinc concentration, with a value of 30.7 ± 9.1 mg/Kg. (Table 1)

Table 1

Selenium and zinc content of pastures consumed by alpacas during three physiological states.
Physiological states /Mineral concentration	Pre-calving	Ten days after calving	Late lactation
Zinc (mg/Kg)	39	32	21
Selenium (mg/Kg)	0.28	0.24	0.32

Selenium in whole blood

The concentration of selenium in whole blood were 53.6 ± 13.9, 56.8 ± 14.4, and 48.2 ± 14.7 ng/ml for female alpacas at pre-calving, ten days after calving, and late lactation respectively (Table 2). For their offspring, selenium in whole blood were 54 ± 14.1 and 53 ± 19.6 ng/ml ten days after calving and late lactation, respectively. No significant differences were found between physiological states concerning selenium concentration in the whole blood of alpacas and their offspring,

Table 2

Whole blood selenium concentration and serum blood zinc concentration of female alpacas during three physiological states and their offspring.
		Physiological states
		Pre-calving	Ten days after calving	Late lactation
Dams (15)	Zinc (µg/ml)	0.154 ± 0.11^b	0.26 ± 0.14 ^a	0.05 ± 0.04 ^c
	Selenium (ng/ml)	53.6 ± 13.9	56.87 ± 14.44	48.2 ± 14.73
Offspring (15)	Zinc (µg/ml)		0.23 ± 0.15 ^a	0.07 ± 0.04 ^b
	Selenium (ng/ml)		54 ± 14.09	53 ± 19.63
Note: Letters with different superscript in rows (a, b, c), indicates significant differences (P < 0.05) between physiological states

Zinc in serum blood

A higher concentration of zinc in the blood serum of female alpacas were observed ten days after calving (0.26 ± 0.2 µg/ml) (P < 0.05), than pre-calving (0.15 ± 0.1 µg/ml) and late lactation (0.05 ± 0.04 µg/ml). On the other hand, the concentration of zinc in blood serum for their offspring were higher ten days after calving (0.23 ± 0.2 µg/ml) (P < 0.05), than late lactation (0.07 ± 0.04 µg/ml) (Table 2).

The concentration of zinc in serum blood was different between the physiological states of female alpacas: ten days after calving with a higher value than pre-calving and late lactation. These differences can be explained because large amounts of zinc (blood serum) are used for the development of the fetus and the production of colostrum and milk, decreasing the reserves of this mineral in the mother (Pavlata et al. 2018). The colostrum can have very high concentration of zinc compared to the blood serum of the mother, indicating a mobilization of zinc from the mother to the calf's first milk (Pavlata et al. 2004). Both processes coincide with what was determined by Badiei et al. (2011), where in addition to what has already been mentioned, adds that one month after calving, the concentration of zinc in blood serum tends to increase again, coinciding with what was determined by Meglia et al. (2004), Yokus and Cakir (2006), and Karatzia et al. (2016). Similar trends were determined by Akhtar et al. (2010), in buffaloes, obtaining that the concentration of zinc in the last third of pregnancy decreases compared to the initial stages of pregnancy and returns to rise during the lactation phase, as in sheep (Aliarabi et al. 2018). The concentration of blood serum zinc from the offspring were not the same between physiological states, lower values were in late lactation compared to ten days after calving. This can be explained due to the consumption habits of the offspring, they are feeding mainly with colostrum (Pavlata et al. 2004), and with a few pastures up to two months of age. When they reached 5 to 6 months of age, they are weaned (FAO 1996); in this study the management of offspring at weaning it occurs in October, after 7 months of birth, coinciding with a greater intake of pastures than milk.

The concentration of zinc determined in the diets was like that found by Ojeda (2020). Likewise, as determined by Espinoza et al. (1982), and Zech and Feuerer (1984), in Bolivia, Burton et al. (2003) in Chile, Judson et al. (1999), and Judson et al. (2011), in Southern Australia. This concentration was adequate during pre-calving, ten days after calving (≥ 30 mg/kg), and marginal during late lactation (20-29.9 mg/kg) (Van Saun and Herdt 2014). According to the assessment of the zinc concentration in pastures, it would be expected that the blood serum during the first two physiological stages should be adequate and during late lactation, it would expect to be deficient, but not so low. Blood serum zinc concentration greater than 0.05 µg/ml was like that obtained by Rosadio et al. (2012), in vicuñas from Peru. However, Semevolos et al. (2013), found a higher concentration of this mineral in the United States, as did Dwyer et al. (2019), in New Zealand and Van Saun and Herdt (2014), in North America. The low concentrations obtained can be attributed to factors such as management (stress, zinc sequestration), and the analysis protocol (elimination of the protein part). However, the evaluation criteria should not only be limited to the individual concentration of the mineral but also consider the concentrations of other minerals, to evaluate the interactions. In the case of zinc, this is less absorbed if there is a higher concentration of sulfur, copper, and iron in the pastures. The first macro mineral affects the availability of zinc in the rumen (Goff 2018), and the second micro mineral if it is in a ratio of 50 to 1 with zinc, will affect the absorption at the level of the intestine (McDowell 2003b; Goff 2018), and the third micro mineral competes for the transport (DMT1) in the same area (intestine) and within the blood (plasma, transferrin) (Georgievski et al. 1982). Using data from a first phase (carried out in some points that cover the highlands of Peru) not yet published, added to what was mentioned above. The sulfur concentration was in the adequate range (0.15–0.20% DM), so its antagonistic role can be ruled out. On the other hand, the iron concentration was excessive (> 400 mg/kg), which could alter the absorption of zinc at the level of the apical membrane of enterocytes and in its transport at the blood plasma level. As for the copper concentration, the average was 14.45 mg/kg, considered adequate, analyzing the copper: zinc ratio, this was 0.47:1, so copper could not intervene in the absorption of zinc at gut level.

On the other hand, the non-variation of selenium in the whole blood of female alpacas during the three physiological states: Pre-calving, ten days after calving, and late lactation, was similar to that determined by Herdt (1995), in llamas, where they found no differences between the concentration of selenium in blood serum, during pregnancy, and after calving, furthermore to the determined by Kachuee et al. (2019), in goats, where they measured whole blood selenium concentration weeks before calving and during calving. As in the female alpacas, in their offspring, no difference was found in whole blood selenium concentration.

About the relation between selenium concentration in pastures and whole blood, the selenium concentration in pastures was like that determined by Judson et al. (1999), and Judson et al. (2011), in Southern Australia. However, it was higher than that determined by Espinoza et al. (1982), in Bolivia. According to Van Saun and Herdt (2014), selenium in pastures is adequate (> 0.2mg/kg), so it would expect that the whole blood selenium is adequate. Whole blood selenium concentration in this study was higher than that found by Ellison (2006), in alpacas bred in New Zealand, and by Judson et al. (1999) and Judson et al. (2011) in alpacas raised in Southern Australia. However, our concentration of selenium in whole blood was lower compared to that determined by Husakova et al. (2014), in alpacas raised in Europe, agreeing with that found by Chicaiza et al. (2016), in Ecuador. Likewise, in other species, such as llamas raised in the United States (Smith et al. 1998), as well in camels (Faye and Seboussi 2009). Considering what is mentioned by Van Saun and Herdt (2014), the concentration of selenium in whole blood obtained in the present study are deficient, the sampled alpacas should show some clinical sign of deficiency (weakness, stiffness of the limbs, white muscle disease, susceptibility to bacterial diseases, metritis), however, none of these signs were observed.

Nonetheless, Judson and Husakova mentioned in both publications that although the values obtained were not that recommended by Van Saun and Herdt, the alpacas did not show clinical signs of deficiency, considering the ranges found adequate for their countries. Perhaps, in our case to confirm this assessment, the interaction of selenium with other minerals should be also considered, for example, selenium may not be absorbed due to modifications at the rumen by sulfur (Fowler 2010; Herdt 2011; Van Saun 2014) and in the small intestine by iron (Semevolo et al. 2013; Mehdi and Dufrasne 2016). As in the case of zinc, comparing selenium concentrations with unpublished data obtained from a first phase, the average sulfur concentration is within the appropriate range (0.15–0.20%), so it would not affect the selenium availability. Regarding the iron concentration, the values were excessive (> 400 mg/kg), iron to be absorbed must be as ferrous (Fe^+ 2), in the diet it is generally found as ferric (Fe^+ 3). In ruminants, at the level of the abomasum, ferric can be reduced to ferrous by the action of acids and at the level of the duodenum by the action of enterocyte ferrireductase (Goff 2018). If there was an excess of iron, this trace mineral in a form ferric could antagonize selenium absorption by forming complexes in the duodenum, which would decrease its absorption.

The concentration of zinc in blood serum of female alpacas varied among physiological states, being higher after ten days after calving (P < 0.05), compared to when they were in pre-calving and late lactation. Selenium concentration in the whole blood of female alpacas doesn´t change among physiological states, having similar values in pre-calving, ten days after calving, and late lactation.

In the case of the offspring, the concentration of zinc in blood serum was higher ten days after calving than late lactation (P < 0.05). On the other hand, no difference was found between ten days after calving and late lactation in whole blood selenium concentration.

For extending understanding of selenium and zinc content in blood of alpacas work should be conducted in pasturing areas with contrasting content of those minerals or simulated in experimental trials. Also, it is advised to take samples from the liver and milk (colostrum and during lactation) to have greater detail and explain the mineral distribution phenomena.

Acknowledgments

Funds for this study were provided by CONCYTEC through a support project (N° 178-2015-FONDECYT) to the Ph.D. program in Animal Science of Universidad Nacional Agraria La Molina.

Statement of Animals Rights

The animal care and experimental procedures used in this study conformed to the regulations and guidelines of the Animals Ethics Committee and the Ethical and Animal Welfare Committee of Universidad Nacional Agraria La Molina (TR. No 0.358-CU-UNALM) and the consent of the owner of alpacas.

Conflict of interest statement

There’s no conflict of interest to public this article.

Funding

Funds for this research were provided by CONCYTEC through a support project (N° 178-2015-FONDECYT) to the Ph.D. program in Animal Science of Universidad Nacional Agraria La Molina.

Competing Interests

The authors have no relevant financial or non-financial interests to disclose.

Authors' contributions

All authors contributed to the study conception and design. Investigation, formal analysis, writing the original draft, resources, software, validation, formal analysis, review, and editing were performed by Sergio Antonio Vargas Mendivil, Carlos Gomez and Mariano Gonzalo Echevarria Rojas. The first draft of the manuscript was written by Sergio Antonio Vargas Mendivil and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Data availability

The datasets generated during the current study are not publicly available but are available from the corresponding author on reasonable request.

Ethics approval

The animal care and experimental procedures used in this study conformed to the regulations and guidelines of the Animal Ethics Committee and the Ethical and Animal Welfare Committee of Universidad Nacional Agraria La Molina.

Consent to participate

Informed consent was obtained from all individual participants included in the study

Consent to publish

The authors participants provided informed consent for publication.

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Blood content of selenium and zinc of female alpacas and their offspring raised in the Andes.

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Abstract

Introduction

Material and Methods

Location

Animals

Sampling and analytical procedure

Statistical analysis

Results

Zn and Se content in pastures

Selenium in whole blood

Zinc in serum blood

Discussion

Declarations

References

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