Effects of Mangifera indica leaf hay on weaned lambs before and after a Haemonchus contortus infection

doi:10.21203/rs.3.rs-2476415/v1

Download PDF

Research Article

Effects of Mangifera indica leaf hay on weaned lambs before and after a Haemonchus contortus infection

https://doi.org/10.21203/rs.3.rs-2476415/v1

This work is licensed under a CC BY 4.0 License

Version 1

posted

You are reading this latest preprint version

The aim of this study was to evaluate the effect of Mangifera indica leaf hay on the control of Haemonchus contortus in lambs, Sixteen crossbred Santa Inês x Dorper lambs were divided into two homogeneous groups. One group was provisioned with 5g of leaf hay /Kg of body weight (BW) and other was not with the hay of this plant.On the 21st day, the animals of both groups were inoculated with infective larvae from H. contortus. The growth, intake and performance for animals were evaluated before and after the nematode infection. Lamb blood parameters were analyzed before the hay administration (day − 14), when the animals receiving the hay and were not-infected (day 7) and when receiving the hay and were infected with H. contortus (day 30). The faeces were collected on day 42, 44, 46, and 48 of the experiment to quantify the eggs per gram of faeces (EPG). The body weight gain was lower when the animals were infected to both groups. In the second period, the animals fed with the hay of M. indica showed higher feed efficiency than the un-treated group. The eosinophil concentrations were higher in the last two periods evaluated. However, the EPG averages were similar between lamb groups. Thus, the inclusion of hay from M. indica leaves in the lamb feed can lead to an improvement in feed efficiency of the animals infected with H. contortus. It can also increase their eosinophil concentration, which in turn could improve tolerance to the nematode infection.

Haemonchosis

sheep production

mango

blood parameters

feed efficiency

The occurrence of gastrointestinal parasites has always been a great challenge for sheep farming. The H. contortus, hematophagous parasite of the abomasum, is considered one the most pathogenic nematode of small ruminants (Lins et al. 2019; Arsenopoulos et al. 2021). The infection by this nematode promotes anemia, inflammation of the abomasum, ascites, submandibular edema, decreased productive performance and, in many cases, death of the ruminants (Naeem et al. 2021).

Conventional treatments use synthetic anthelmintics; however the indiscriminate use of the product has selected H. contortus populations resistant to several anthelmintic groups (Arsenopoulos et al. 2021). In Brazil, studies in different regions have conducted research on populations of the multi-resistant nematodes (Nagata et al. 2019; Amaral et al. 2021; Amaral et al. 2021,Viana et al. 2021).

Alternative control methods have been developed to control resistant populations, such as the use of biological control with fungi (Braga et al. 2020) or even through the use of vaccines (Teixeira et al. 2019; Ehsan et al. 2020). However, most research has focused on the effects of plant use (Fonseca et al. 2014; Kuiseu et al. 2021). Studies have demonstrated that the tannins present in different groups of plants may promote larvicidal (Ahmed; Al-Joubori, 2020) and ovicidal effects (Mravcaková et al. 2020). Although some tannins showed undesirable feed effects, Inácio et al. 2021 performed an in vivo study with sheep treated with extracts of Mauritia flexuosa leaves containing tannin; this approach showed anthelmintics effects without compromising the productivity of animals.

Mangifera indica L. has antioxidant, analgesic, anti-inflammatory effects, immunomodulatory action, activity in the control of chronic bronchitis, dysentery and intestinal hemorrhage in humans, in addition to diuretic activity and stimulation of milk production (Gonzales et al. 2007). Nery et al. 2012 used fresh M. indica juice at 74 g/mg body weight (BW) to control gastrointestinal nematodes; he observed that the number of eggs per gram of faeces (EPG) was reduced by 53%. However, the effects of adding leaves, which are available all year, to the diet of healthy or H. contortus-infected sheep are not known. In this study we aimed to evaluate the influence of hay from the leaves of M. indica on weaned lambs before and after the H. contortus infections.

2.1 Collection and preparation of hay from mango leaves

The leaves of M. indica var Ubá were collected in March and April, at the Montes Claros, North of Minas Gerais, Brazil. Damaged or deteriorated leaves were discarded and the selected material was washed and dehydrated in an oven with regulated air circulation (TE 394/4, Tecnal Scientific Equipment Tecnal, Piracicaba, SP, Brazil) at 40ºC ± 5 until it reached a constant weight. The leaves were then ground and stored in paper packages away from light (Morais-Costa et al. 2015). The subsamples were and then placed in the oven to determine dry matter (DM) at 105ºC according to Association of Official Analytical Chemists AOAC (1997).

2.2 Animals And Diets Evaluated

All procedures were performed in accordance with the princi- ples of animal experimentation approved in the 120/2016 protocol of the Ethics Committee on the use of animals (CEUA) of the Federal University of Minas Gerais, Brazil. Sixteen recently weaned crossbred Santa Inês x Dorper female lambs approximately 3.5 months of age with an average body weight (bw) of 15 ± 2.73 kg were included in this study. During the adaptation period, albendazole (10 mg/g of body weight) was administered to the lambs and eggs per gram (EPG) of faeces tests were performed on three consecutive days to ensure that the lambs were free of nematodes. The animals were then randomly divided into two groups.

The strains of leaves were evaluated according to AOAC (1997) for dry matter (DM), crude protein (CP) ether extract (EE). Neutral detergent fiber (NDF) analyses were performed according to procedures described by Van Soest et al., (1991). Acid detergent fiber was evaluated by AOAC, (1997) using ANKOM200 Fiber Analyzer (ANKOM Technology Corporation, Fairport, Nova York, EUA).

The proportion of ingredients and their chemical compositions are shown in Table 1. One group was treated with mango leaf hay (MLH) at 5 g/kg/bw instead of corn silage and the other was not treated, representing the negative control. The diets were formulated according to the National Research Council (2007), to contain 50% roughage (corn silage) and 50% concentrate based on corn bran, soybean meal, limestone, and bi-calcium phosphate (Table 2). The forage and concentrate were mixed in the troughs and fed to the animals twice a day, in two equal portions. One portion was offered in the morning (8:00 am) and the other in the afternoon (4:00 pm). The amount of feed provided was calculated to allow for 10% leftovers.

Table 1

Chemical composition of the ingredients (g/Kg of dry mater) used in the experimental diets of the sheep evaluated
Ingredients	mango leaf hay	corn silage	ground corn grain	Soybean meal
Dry matter	908.7	272.9	876.4	886.1
Mineral matter	85.8	59.0	16.1	63.9
NDF	371.7	484.4	139.1	122.2
ADF	242.1	220.0	54.4	101.3
EE	76.5	23.8	40.2	34.5
CP	85.2	110.1	90.5	487.8
Note: crude, protein (CP) ether extract (EE). Neutral detergent fiber (NDF), Acid detergent fiber (NDF)

Table 2

Ingredients in experimental diets with or without *Mangifera indica* leaf hay (MLH) (g/kg of matter and chemical composition according to treatments.
Ingredients (%DM)	Diet without MLH	Diet with the MLH
Corn silage	500.0	380.0
Corn	340.0	329.0
Soybean meal	140.0	148.0
Bi-calcium phosphate	20.0	20.0
Mango leaf hay	00.0	123.0

2.3 Experimental Design

For adaptation, the sheep were confined to individual stalls for seven days. They were fed corn silage to help them adapt to the place, when were also given 14 days to adapt to their diets. The experiment was carried out in a split-plot (two lamb groups and two periods). The first period comprised 21 days with healthy animals free from nematode infection. In the second period, the animals were infected with 800 infective larvae of H. contortus /kg/bw and evaluated over 14 days.

2.4 Evaluated Variables

The morphometric measures, DM ingestion, and performance (daily body weight gain, feed conversion e feed efficiency) of the animals were evaluated weekly during the adaptation and then the experimental period. The animals were inspected for possible clinical or behavioral alterations according to the procedure described by Dirksen (1993).

Once a week, the ewe lambs were weighed on a digital scale. For each period, morphometric measurements (height at withers, croup height, chest height, body length, croup length, croup width, chest circumference, neck circumference, and chest width) were evaluated upon a straight floor with the aid of a tape measure.

For the analysis of hematological parameters, blood was collected from the lambs thrice over the three periods; in period one (before the animals started to receive the diet with the treatment, during the adaptation period, day − 14),period two (the animals receiving the treatment and not infected, day 7) and period three (animals finishing the treatment period and already infected with H. contortus, day 30). Blood was collected by jugular vein puncture after asepsis with 1% PVP-iodine I, using vacuum tubes. Tubes containing ethylenediamine tetraacetic acid (EDTA) were sent for analysis of blood counts in electronic counting equipment (BC 2,800 Vet, Mindray Medical International LTDA, China). The analyses was performed in split plots (two treatments and three periods), with eight replications. The data were log-transformed and evaluated at 5% significance by the Scott-Knott test.

The egg reduction test in ewes' faeces was according to the methodology adapted from Coles et al. (1992). Faeces were collected on days 42, 44, 46, and 48 of the experiment, after the nematode attained maturity. EPG means were obtained individually using the modified McMaster technique with two replicates per animal (Gordon; Whitlock, 1939). Fecal samples from all collections were used for coproculture to confirm the nematode genus (Ueno; Gonçalves, 1988). The experiment was carried out in a split-plot design (plots represented by the two treatments and the sub-plots by the four analysis periods). The EPG data obtained were log10 (x + 10)-transformed and submitted to an analysis of variance. The means were compared by Scott-Knott tests, at 5% significance. The formula adapted from Coles et al. (1992) was employed to determine the percentage reduction of eggs in faeces: % Efficacy = 100 x (1 – mean EPG treated group / mean EPG control group).

Statistical analysis

After normality and homogeneity analyses, the data were analyzed in a split plot considering the two treatments and two periods (as measures repeated in time) and the interaction between the treatments and periods, by the statistical model:

Yijk = m + Ai + Bj + Ck + (A x C) ik + ebijk

Yijk was the observation; m was the overall mean; Ai was the effect of treatments (i = 2); Bj was the random effect of the animals (n = 8); Ck was the effect of the periods (k = 2); (A x C) ik was the fixed effect of the interaction between treatments and periods and ebijk was the effect of experimental error. The blood parameters from the two treatments and three periods were also evaluated in a split plot. The means were evaluated and compared by the Scott–Knott test, at a significance level of 5%, and the SAEG 9.1 statistical package was used.

3.1 Animal performance

In the initial period (uninfected), animals fed with the MLH additive had a higher dry matter intake (DMI) than those in the control group (P < 0.05, Table 3). In the second period, the infected animals in the control group had a higher DMI compared to the first period while the animals treated with MLH showed a decrease in DMI (P < 0.05, Table 3). The interaction between treatments and periods was significant (P < 0.05).

Table 3

Performance of ewes fed on a diet that included *Mangifera indica* leaf hay (MLH) before and after *Haemonchus contortus* infection.
	Before nematode infection		After nematode infection
Parameters	Control	MLH	Control	MLH
DMI (g/day)	0.69 ± 0.08 ^Bb	0.90 ± 0.12^Aa	1.06 ± 0.16 ^Aa	0.62 ± 0.07 ^Bb
DBWG (g/day)	0.15 ± 0.05 ^Aa	0.14 ± 0.04 ^Aa	0.10 ± 0.02^Ab	0.08 ± 0.01^Ab
Feed conversion	6.23 ± 3.37 ^Aa	7.36 ± 2.56 ^Aa	9.97 ± 0.59 ^Aa	7.57 ± 1.51 ^Aa
Food efficiency	0.22 ± 0.20 ^Aa	0.18 ± 0.24^Aa	0.10 ± 0.005^Bb	0.14 ± 0.03 ^Aa
Note: Different capital letters indicate the difference between lamb groups in the same periods and different lower case letters indicate difference between periods in the same lamb groups by Scott Knott test (P < 0.05).
DMI = Averages of total dry matter intake in grams per day.
DBWG = Averages of daily body weight gain in grams per day.
Feed conversion = DMI/DPVD.
Food efficiency = DPVD/DMI.

There was no significant difference in body weight gain (BWG) between the lambs groups evaluated. However, this gain was lower in the second period for both groups, when the animals were infected (P < 0.01, Table 3). There was no difference in feed conversion (FC) for both lamb groups and periods evaluated (P > 0.05). However, in the second period, when the ewe lambs were infected, the group treated with MLH presented higher FE than the un-treated group (P < 0.05). Analyzing the periods evaluated, the control group significantly reduced the FE, while the treated group maintained it (Table 3). The interaction between treatments and periods was significant (P < 0.05).

For the average daily gain of withers height, croup height, chest height, body length, and rump length, no significant differences were detected between treatments in any of the evaluated periods (Table 3). In the second period, after upon infecting the animals, these growth parameters were reduced for both groups of sheep evaluated (P < 0.01). For the average daily gains in rump width, chest circumference, neck circumference, and breast width, no significant differences were detected between the lamb groups and periods evaluated (Table 4).

Table 4

Mean daily gain in morphometric measurements (cm) of lambs (fed or unfed) *Mangifera indica* leaf hay (MLH) before and after infection by *Haemonchu contortus*.
Morphometric measurements	Before nematod infection		After nematode infection
	Control	MLH	Control	MLH
Height at withers	0.21 ± 0.08 ^Aa	0.18 ± 0.06 ^Aa	0.09 ± 0.02 ^Ab	0.12 ± 0.06 ^Ab
Rump height	0.28 ± 0.07 ^Aa	0.22 ± 0.09 ^Aa	0.12 ± 0.04 ^Ab	0.13 ± 0.06 ^Ab
Chest height	1.11 ± 0.05 ^Aa	1.09 ± 0.03 ^Aa	0.09 ± 0.08 ^Ab	0.05 ± 0.02 ^Ab
Body length	2.56 ± 0.16 ^Aa	2.51 ± 0.07 ^Aa	0.17 ± 0.03 ^Ab	0.13 ± 0.05 ^Ab
Rump length	0.20 ± 0.07 ^Aa	0.16 ± 0.09 ^Aa	0.09 ± 0.04 ^Ab	0.05 ± 0.04 ^Ab
Rump width	0.14 ± 0.03 ^Aa	0.12 ± 0.05 ^Aa	0.10 ± 0.06 ^Aa	0.08 ± 0.03 ^Aa
Chest circumference	0.25 ± 0.08 ^Aa	0.25 ± 0.13 ^Aa	0.17 ± 0.05 ^Aa	0.18 ± 0.08 ^Aa
Neck circumference	0.14 ± 0.03 ^Aa	0.12 ± 0.07 ^Aa	0.11 ± 0.05 ^Aa	0.37 ± 0.42 ^Aa
Breast width	0.11 ± 0.03 ^Aa	0.09 ± 0.05 ^Aa	0.14 ± 0.03 ^Aa	0.08 ± 0.04 ^Aa
Note: Morphometric measurements expressed in centimeters. Different uppercase letters indicate differences between lamb groups and different lowercase letters indicate differences between periods by the Scott Knott test (P ≤ 0.05).

3.2 Blood Parameters And Fecal Nematode Egg Counting

The concentration of red blood cells was not influenced by the treatment. The concentration was lower in the second period for both lamb groups (P < 0.05, Table 5). The concentration of hemoglobin, hematocrit, and platelets was not influenced by the treatments. However, in both groups, concentrations were lower in the first period (P < 0.01). For both lamb groups, the mean corpuscular hemoglobin and mean corpuscular volume values were higher in the second period. However, corpuscular hemoglobin concentrations and erythrocyte distribution amplitude increased with the age of lambs (P < 0.01, Table 5).

Table 5

Hemogram of uninfected female lambs before day 14 and on day 7 upon treatment with *Mangifera indica* leaf hay (MLH); hemogram of female lambs infected with *Haemonchus contortus* (treated or untreated) with MLH day 30.
Lambs groups	Before nematode infection		After nematode infection
	Day − 14	Day 7	Day 30	Reference ^a	CV (%)
Red blood cells (Millions/mm)
Not treated	5.26	4.24	5.46	9–15	18,02
(MLH)	5.84	4.68	5.726	9–15	18,02
Mean	5.05a	4.46b	5.59a
Hemoglobin (g/dL)
Not treated	12.31	14.86	16.02	9–15	17,56
(MLH)	12.12	15.32	16.14	9–15	17,56
Mean	12.21b	15.09a	16.08a
Hematocrit (%)
Not treated	36.94	42.92	44.34	27–45	17,36
(MLH)	37.3	43.76	44.06	27–45	17,36
Mean	37.12b	43.34a	44.2a
Mean corpuscular hemoglobin (pg)
Not treated	23.53	35.44	30.92	8–12	5,84
(MLH)	20.79	33.66	28.23
Mean	22.16c	34.55a	29.57b
Corpuscular hemoglobin concentration (g/l)
Not treated	33.30	34.67	36.14	31–34	14,38
(MLH)	32.50	35.14	36.62	31–34	14,38
Mean	32.9c	34.90b	36.38a
Mean corpuscular volume (fl)
Not treated	70.63	102.22	85.43	28–40	7,33
(MLH)	64.02	95.28	77.16
Mean	67.32c	98.75a	81.29b
Range of distribution of erythrocytes - RDW (%)
Not treated	13.32	16.98	18.94	*	17,59
(MLH)	13.56	17.12	18.08	*	17,59
Mean	13.44c	17.05b	18.51a
Platelets (µl)
Not treated	343800	368000	386800	250–750	6,54
(MLH)	310000	457000	403600	250–750	6,54
Mean	326900b	412500a	395200a
Capital letters in the columns and lowercase letters in the rows differ significantly (P ≤ 0.05). Reference values for healthy sheep (PUGH, 2004). * Values not reported in the consulted literature.

The values of leukocytes, lymphocytes, and neutrophils were not influenced by treatments, being lower at first period evaluated for both groups lambs (P < 0.01, Table 6). Monocyte concentrations showed no influence of treatments and periods (P > 0.05). There was a significant interaction between treatments and periods evaluated for eosinophil counts. The concentrations of these were higher for the treated group in the last two periods evaluated in relation to the control group (P < 0.01, Table 6). The mean EPG of both lamb groups and periods was similar (P > 0.05) and there was no evidence of EPG reduction in the group treated with MLH (Table 7).

Table 6

Leukogram of uninfected female lambs treated with *Mangifera indica* leaf hay (MLH) before day 14 and on day 7. Leukogram of infected female lambs with *Haemonchus contortus* (treated or untreated) with MLH (day 30).
Lambs groups	Before nematode infection		After nematode infection
	Day − 14	Day 7	Day 30	Reference ^a	CV (%)
Leukocytes (mm³)
Not treated	5460	8400	8360	4000–12000	3,09
MLH	6200	8820	8480	4000–12000	3,09
Mean	5830b	8610a	8420a
Lymphocytes (mm³)
Not treated	1836,2	2285,0	2570,2	2.000–9.000	3,42
MLH	1898,6	2673,6	2804,6
Mean	1867,4b	2479,3a	2687,4a
Monocytes (mm³)
Not treated	151Ac	197,8Bb	207,2Ba	0–750	7,02
MLH	149,8Ac	247,8Ab	252,4Aa	0–750	7,02
Eosinophils (mm³)
Not treated	157,6Ab	174,8Ba	112,0Ba	0 -1000	31,94
MLH	84,4Bb	277,0Aa	166,2Ab	0 -1000	31,94
Rods neutrophils (mm³)
Not treated	23,8	0.0	0.0	*	24,53
MLH	0.0	0.0	0.0	*	24,53
Segmented neutrophils (mm³)
Not treated	3376,2	5650,2	5420,4	400 a 6.000	3,60
MLH	3992,2	5723,8	5311,0
Mean	3684,2b	5687ª	5365,7a
Total neutrophils (mm³)
Not treated	3388,4	5650,2	5420,4
MLH	3992,2	5723,8	5311,0
Mean	3690,3b	5687ª	5365,7a
Capital letters in the columns and lowercase letters in the lines differ significantly (P ≤ 0.05). Reference intervals for healthy sheep (PUGH, 2004). * Values not reported in the consulted literature

Table 7

Mean values of eggs per gram of faeces of *Haemonchus contortus* from lambs (treated or untreated) with *Mangifera indica* leaf hay (5g/kg/bw)
Collection period (days)
Lambs groups	42	44	46	48	Mean
Not treated	437.5 ± 120.5	756.2 ± 320.5	956.2 ± 230.3	825.0 ± 320.3	743.7 ± 220.3
M. indica leaf hay	725.0 ± 121.3	1162.5 ± 220.3	787.5 ± 128.3	1006.2 ± 380.3	920.3 ± 201.5

4.1- Animal performance

In this study, weight gain, FC, and FE were similar between lamb groups after weaning (before the H. contortus infection), indicating that MLH could represent a nutritional alternative. Additionally, the inclusion of this hay could contribute to reducing diet costs in relation to roughage, since the partial replacement of corn silage by MLH did not lead to performance losses and the leaves of this plant are available in many rural properties during all the seasons.

After nematode infection, the improved FE of lambs treated with MLH can potentially be attributed to the presence of phenolic compounds such as flavonoids. These metabolites may positively contribute to the productive performance and health of ruminants (Olagaray; Bradford, 2019). Sheep supplemented with flavonoids from mulberry leaves (Morus spp.), (Ma et al. 2017) presented changes in ruminal microbial ecology with an increase in the total bacterial population, mainly fibrinolytic bacteria, decreased protozoa, methanogenic individuals, and consequently, the production of methane. However, the presence of condensed tannins may decrease the dry matter intake (Min; Soleiman, 2018).

The average DMI (0,802 Kg) observed in this study was close to that recommended by NRC (2007), which is 0.800 Kg for lambs with an average BW of 20 Kg. The study carried out by Carvalho et al. (2021) compared the effect of four diets on Dorper sheep infected with H. contortus and Trichostrongylus colubriformis. The energy levels and protein in the diet influenced the H. contortus infection; the animals that received higher metabolizable protein and energy showed a lower incidence of hipoproteinemia. In cases of infection with these nematodes, diets with balanced levels of proteins protected from rumen degradation would be recommended to improve EF, since it could provide greater amounts of amino acids and peptides that would be absorbed in the small intestine (Atiba et al. 2021).

Morphometric measures were not influenced by the MLH inclusion; however in the second period, after infection, these growth parameters were reduced in both groups as a result of inflammatory effects on the abomasal mucosa parasitized with H. contortus. Similarly, Araújo Filho et al. (2007), reported that with the exception of thorax circumference and breast width, diet did not have a significant effect on morphometric measurements of feedlot sheep; the exceptions could be attributed to greater tissue deposition, and were observed in higher energy diets. This, however, was not the case in our study, given that the diets had similar levels of NDF.

In blood analyses, the higher eosinophil concentration observed in the lambs that ingested MLH in the last two evaluated periods could be attributed to a stronger reaction to the nematode infection triggered by MLH metabolites such as the flavonoids, which are known to have antibacterial, antiparasitic, immunomodulatory, and anti-inflammatory activities in addition to being potent antioxidants and enzimatic inhibitors (Middleton et al. 2000; Wang et al. 2007; Sulsen et al. 2007; Wang et al. 2010; Sandhar et al. 2011).

Eosinophils are important for the control of gastrointestinal nematode infections since these cells may reduce L3 to L4 of the H. contortus evolution on abnormal mucosa (Balic et al. 2006). These cells take action in an integrated way along with cytokines, chemokynes, and other cells of the immune system (Huang; Appleton, 2016).

A study that evaluated the action potential of eosinophils in vitro from the blood of H. contortus-infected sheep found that esosinophils promoted drastic reduction of L3 incubated with eosinophil extracted from infected animals. After incubation, the larvae were transferred intra-abomasum and the majority was not established in the host (Terefe et al. 2007). Thus, in our study the increase in the number of eosinophils in animals treated with MLH could also have contributed to parasite reduction in a chronic phase of haemonchosis, which should be evaluated in future studies.

In cases of intense parasitism in susceptible animals, there is an increase in TNFα and IL1β. This can promote to an increase in the inflammatory response. In resistant animals, an increase in IL-33 expression is observed as a reflection of the polarized Th2 response (Toscano et al. 2019). Similarly, it was observed by Jacobs et al. (2018) that IL-4 is important in activating of Th2 response, and is therefore related to the establishment of infection. Furthermore, other Th2 mechanisms are activated in helminth infections including IL-13 and IL-25 (Albuquerque et al. 2019; Toscano et al. 2019; Ehsan et al. 2020).

A study evaluated resistant Santa Inês sheep and susceptible Ile France sheep infected with H. contortus that were submitted to a suppressive treatment or other target selective treatment. The researchers reported different mechanisms involved in the response to infection, including early and robust eosinophilia and higher circulating levels of IgA in resistant Santa Inês sheep, denoting how the breed factor becomes important in parasitism (Albuquerque et al. 2019).

In this study, there were no significant differences between the means of EPG for groups of animals and periods that represented the initial phase of haemonchosis. However, in future research, this analysis should also be considered for a longer period, given that the animals treated with MLH demonstrated an increase in the concentration of eosinophils and could possibly reduce the number or viability of adult nematodes during a chronic phase of the disease.

Alternatively, a study that evaluated the inclusion of banana leaf hay (Magdeleine et al. 2010) did not report a significant effect on immunity against H. contortus. However, when treated with this hay after 21 days of infection with 5000 larvae from H. contortus, the authors observed lower reduction of EPG compared to un-treated animals. Another study by El-Sherbini, Osman (2013) evaluated M. indica L. and reported that compounds present in immature fruits, including the flavonoids, inhibited the larval development of the nematode Strongyloides stercoralis. When analyzing the tannins presents in leaves of Mauritia flexuosa, Inácio et al. (2021) detected 33.23% ± 2 of condensed tannins. When they treated the lambs with an aqueous extract of this palm at 62.1 mg/kg de BW for sheep infected with H. contortus, they found that the anthelmintic efficacy was 54.57% after 14 days.

4.2 Conclusion

The inclusion of M. indica leaf hay at 5g/Kg BW does not compromise morphometric development, and the weight gain of weaned lambs. Furthermore, the inclusion of this additive promotes improved feed efficiency and increases the concentrations of eosinophils when these animals are infected with H. contortus.

Acknowledgement

To the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), to the financial support and scholarships from Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG), and Pesquisa Pró-Reitoria da Universidade Federal de Minas Gerais (PRPq-UFMG).

Conflict of interest statement

The authors of this manuscript have no financial or personal relationship with individuals or organizations that could influence or bias the content of the paper.

Author contribution

Conceptualization: ERD; methodology: KMM, FM-C, VSMJ, LCG; ; investigation: ERD, KMM, FM-C, VSMJ; data curation: ERD, KMM, FM-C; writing—original draft preparation: KMM, ERD, ASC; ; review and editing: EFSA, AJS, KMM, ERD, ASC; supervision: ERD, All authors have read and agreed to the manuscript’s current published version; Data Availability Statement All Data are available upon request

Ethics approval

In this study all procedures were performed in accordance with the principles of animal experimentation approved in the 120/2016 protocol of the Ethics Committee on the use of animals (CEUA) of the Federal University of Minas Gerais, Brazil.

Funding

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES Brazil, grant 001), Fundeci Bando do Nordeste do Brazil – 0001)

Data availability

All data generated or analyzed during this study are included in this published article.

Ahmed, S. and Al-Jubori, A-A.,2020. Garlic (Allium sativum) and clove (Syzygium aromaticum) as alternative treatments for the control of Haemonchus contortus in sheep. Plant Archives, 20, 106-109.
Albuquerque, A. C. A., Bassetto, C. C., Almeida, F. A., Hildersley, K. A., McNeilly, T. N., Britton, C., Amarante, A. F. T., 2019. Differences in immune responses to Haemonchus contortus infection in the susceptible Ile de France and the resistant Santa Ines sheep under different anthelmintic treatments regimens. Veterinary Research, 50, 1-12. doi:10.1186/s13567-019-0722-3
Amaral, R. L. G., Guerra, N. R., Santana, I. M., Chicoy-Ramirez, Y., Arenal, A., Lima, M. M., Alves, L. C., Molento, M. B., Faustino, M. A. G. 2021. Diagnóstico molecular da resistência de Haemonchus contortus ao grupo dos benzimidazóis em ovinos e caprinos no estado de Pernambuco, Brasil. Archives of Veterinary Science, 26, 59-71.
AOAC - Association of Official Analytical Chemist, 1997. Official methods of analysis 16th edition AOAC, Gaithersburg, MP. AOAC, Gaithersburg, MP.
Arsenopoulos, K. V., Fthenakis, G. C., Katsarou; E. I., Papadopoulos, E. 2021. Haemonchosis: A Challenging Parasitic Infection of Sheep and Goats. Animals, 11, 363. doi:10.3390/ani11020363
Atiba, E. M., Li, Y., Cheng, W., Li, Y., Sun, N.,Sun, Z.; Zhong Q. 2021. Dietary protein degradability effect on performance of lambs experimentally infected with Haemonchus contortus and Trichostrongylus colubriformis. Tropical Animal Health and Production, 53, 1-9. doi:10.1007/s11250-021-02822-z
Balic, A., Cunninghan, C.P., Meeusen, E.N.T. 2006. Eosinophil interactions with Haemonchus contortus larvae in the ovine gastrointestinal tract. Parasite Immunology, 28, 07-115.
Braga, F. R., Ferraz, C. M., Silva, E. N., Araujo, J. V. 2020. Efficiency of the Bioverm® (Duddingtonia flagrans) fungal formulation to control in vivo and in vitro of Haemonchus contortus and Strongyloides papillosus in sheep. 3 Biotech. 10, 1-5. doi:10.1007/s13205-019-2042-8
Carvalho, N., Neves, J. H., Pennacchi, C. S., Castilhos, A. M., Amarante, A. F. T. 2021. Performance of lambs under four levels of dietary supplementation and artificially mix-infected with Haemonchus contortus and Trichostrongylus colubriformis. Revista Brasileira de Parasitologia Veterinária, 30, 1-9. doi:10.1590/S1984-29612021010
Coles, G. C., Bauer, C., Borgsteede, F. H., Geerts, S., Klei, T. R., Taylor, M. A., Waller, P. J. 1992. World Association for the Advancement of VETERINARY PARASITOLOGY (WAAVP) – methods for detection of anthelmintic resistance in nematodes of veterinary importance. Veterinary Parasitology, Amsterdam, 44, 35–44.
Ehsan, M., HU, R.-S., Liang, Q-L., Hou, J-L., Xiaokai, S., Yan, R.., Zhu, X.-Q., Li, X. 2020. Advances in the Development of Anti-Haemonchus contortus Vaccines: Challenges, Opportunities, and Perspectives. Vaccines, 8, 555. doi:10.3390/vaccines8030555
El-Sherbini, G. T., Osman, S. M. 2013. Anthelmintic activity of unripe Mangifera indica L. (Mango) against Strongyloides stercoralis. International Journal of Current Microbiology and Applied Sciences, 2, 401-409.
Dirksen, G. Sistema digestivo. In: Dirksen, G.; Gründer, H. D.; Stöber, M. (Eds.). Rosenberger: Exame clínico dos bovinos. Rio de Janeiro: Guanabara-Koogan, 1993. p.167-169.
Filho, J. T. A., Costa, R. G., Fraga, A. B., Sousa, W. H., Neto, S. G., Batista, A. S. M., Cunha, M. G. G. 2007. Efeito de dieta e genótipo sobre medidas morfométricas e não constituintes da carcaça de cordeiros deslanados terminados em confinamento. Revista Brasileira de Saúde e Produção Animal, 8, 394-404.
Fonseca, L. D., Vieira, T. M., Lázaro, S. F., Silva, M. L. F., Ferreira, A. V. P., Bastos, G. A., Morais-costa, F., Martins, E. R., Duarte, E. R.. 2014. Eficácia in vitro de extratos aquosos de plantas no controle de nematódeos gastrintestinais de bovinos. Acta Scientiae Veterinariae, 42, 1-8.
González, J. E., Rodríguez, M. D., Rodeiro, I., Morffi, J., Guerra, E.., Leal, F., García, H.., Goicochea, E.., Guerrero, S., Garrido, G., Delgado, R.., Nuñez-selles, A. J. 2007. Lack of in vivo embryotoxic and genotoxic activities of orally administered stem bark aqueous extract of Mangifera indica L. (Vimang®). Food and Chemical Toxicology, 45, 2526-2532.
Gordon, H. M., Whitlock, H. V. 1939. A new technique for courting nematode eggs in sheep faeces. Journal of Council of Science, Industry and Research, 12, 50-52.
Huang, L., Appleton, J. A. 2016. Eosinophils in Helminth Infection: Defenders and Dupes. Trends in Parasitology, 32, 798-807. doi:10.1016/j.pt.2016.05.004
Inácio, M. F., Morais-costa, F., Cardoso Filho, O., Martins, I. P., Martins Júnior, V. S., Duarte, E. R.., Arrudas, S. R.., Nunes, Y.. R. F. 2021. Leaves of Mauritia flexuosa L. f. (Arecaceae) is effective in vitro and in vivo in the control of Haemonchus contortus. Research, Society and Development, 10, 1-11. doi:10.33448/rsd-v10i12.20493
Jacobs, J. R., Greiner, S. P., Bowdridge, S. A. 2018. Impaired interleukin-4 signalling promotes establishment of Haemonchus contortus in sheep. Parasite Immunology, 40, e12597 doi:10.1111/pim.12597
Kuiseu, J., Zinsou, F. T. A., Olounlade, P. A., Alowanou, G. G., Adenile, A. D., Dansou, C. C., Hounzangbe-Adote, S., Babayemi, O. J., Edohr, P. A. 2021. Prevalence, effects and alternative control methods of Haaemonchus contortus in small ruminants: A review. Journal of Veterinary Medicine and Animal Health, 13, 84-97. doi:10.5897/JVMAH2020.0868
Lins, J. G. G., Rodrigues, S. D., Marques, A. V. M. S. 2019. Prevalence of gastrointestinal helminths in sheep raised in intermediary geographical region of Paraiba state. Veterinária e Zootecnia, 26, 1-9. doi:10.35172/rvz.2019.v26.358
Lins, J. G. G., Almeida, F. A., Amarante, A. F. T. 2020. Haematological variables of Santa Ines and Ile de France suckling lambs: Influence of Haemonchus contortus infection. Pesquisa Veterinária Brasileira, 40, 882-91. doi:10.1590/1678-5150-PVB-6618
Ma, T., Chen, D-D., Tu, Y., Zhang, N-F.; Si, B-W., Diao, Q-Y. 2017. Dietary supplementation with mulberry leaf flavonoids inhibits methanogenesis in sheep. Animal Science Journal, 88, 72-78. doi:10.1111/asj.12556
Marie-Magdeleine, C., Boval, M.; Philibert, L., Borde, A., Archimède, H. 2010. Effect of banana foliage (Musa paradisiaca) on nutrition, parasite infection and growth of lambs. Livestock Science, 131, 234-239. doi:10.1016/j.livsci.2010.04.006
Middleton, E., Kandaswami, C., Theoharides, C. 2000. The effects of Plant Flavonoids on Mammalian Cells: Implications for Inflamamation, Heart Disease, and Cancer. Pharmacological Reviews, 52, 673 – 751.
Min, B. R., Solaiman, S. 2018. Comparative aspects of plant tannins on digestive physiology, nutrition and microbial community changes in sheep and goats: A review. Journal of Animal Physiology and Animal Nutrition, 102, 1181-1193. doi:10.1111/jpn.12938
Morais-Costa, F., Soares, A. C., Bastos, G. A., Nunes, Y. R., Geraseev, L. C., Braga, F. C., Lima, W. D. S., Duarte, E. R. 2015. Plants of the Cerrado naturally selected by grazing sheep may have potential for inhibiting development of Haemonchus contortus larva. Tropical Animal Health Production, 47, 1321-1328. doi:10.1007/s11250-015-0866-8
Mravcaková, D., Komáromyová, M., Babják, M., Dolinská, M. U., Königová, A., Petric, D., Cobanová, K., Slusarczyk, S., Cieslak, A., Várady, M., Váradyová, Z. 2020. Anthelmintic Activity of Wormwood (Artemisia absinthium L.) and Mallow (Malva sylvestris L.) against Haemonchus contortus in Sheep. Animals, 10, 219. doi:10.3390/ani10020219
Naeem, M., Iqbal, Z., Roohi, N. 2021. Ovine haemonchosis: a review. Tropical Animal Health and Production, 53, 1-11. doi:10.1007/s11250-020-02439-8
Nagata, W. B., Panegossi, M. F. C., Bresciani, K. D. S., Gomes, J. F., Kaneto, C. V., Perri, S. H. V. 2019. Resistance of gastrointestinal nematodes to five different active principles in sheep infected naturally in São Paulo State, Brazil. Small Ruminant Research, 172, 48-50. doi:10.1016/j.smallrumres..10.001
National Research Council - NRC. Nutrient requirements of small ruminants: Sheep, goats, cervids, and new word camelids. Washington, D.C: National Academy Press, 2007. 384p
Nery, P. S., Nogueira, F. A., Oliveira, N. J. F., Martins, E. R., Duarte, E. R. 2012. Efficacy of extracts of immature mango on ovine gastrointestinal nematodes. Parasitology Research, 111, 2467–2471. doi:10.1007/s00436-012-3017-4
Olagaray, K. E., Bradford, B. J. 2019. Plant flavonoids to improve productivity of ruminants – A review. Animal Feed Science and Technology, 251, 21-36. doi:10.1016/j.anifeedsci.2019.02.004
Sandhar, H. K., Kumar, B., Prasher, S., Tiwari, P., Salhan, M., Sharma, P. 2011. A Review of Phytochemistry and Pharmacology of Flavonoids. Internationale Pharmaceutica Sciencia, 1, 25 – 41.
Sulsen, V. P., Cazorla, S. I., Frank, F. M., Redko, F. C., Anesini, C. A., Coussio, J. D., Maldchiodi, E. L., Martino, V. S., Muschietti, L. V. 2007. Trypanocidal and Leishmanicidal Activities of Flavonoids from Argentine Medicinal Plants. American Society of Tropical Medicine and Hygiene, 654-659.
Teixeira, M., Matos, A. F. I. M., Albuqurque, F. H. M. A., Bassetto, C. C., Smith, W. D., Moteiro, J. P. 2019. Strategic vaccination of hair sheep against Haemonchus contortus. Parasitology Research, 118, 2383-2388. doi:10.1007/s00436-019-06367-x
Terefe, G., Grisez, C., Prevot, F., Bergeaud, J. P., Dorchies, P., Brunel, J. C., François, D., Fourquaux, I., Jacquiet, P. 2007. In vitro pre-exposure of Haemonchus contortus L3 to blood eosinophils reduces their establishment potential in sheep. Veterinary Research, 38, 647-654.
Toscano, J. H. B., Okino, C. H., Santos, I. B., Giraldelo, L. A., Haehling, M. B. V., Esteves, S. N., Chagas, A. C. S. 2019. Innate Immune Responses Associated with Resistance against Haemonchus contortus in Morada Nova Sheep. Journal of Immunology Research, 2019, 1-11. doi:10.1155/2019/3562672
Ueno, H., Gonçalves, P. C. 1988. Manual para diagnóstico das helmintoses de ruminantes. Tokyo: Japan International Cooperation Agency
Van Soest, P.J., 1965. Symposium on factors influencing the voluntary intake of herbage by ruminants: voluntary intake in relation to chemical composition and digestibility. J. Anim. Sci. 24, 834–843. https://doi.org/10.2527/jas1965.243834x.
Viana, M. V. G., Silva, Y. H., Martins, I. V. F., Scott, F. B. 2021. Resistance of Haemonchus contortus to monepantel in sheep: first report in Espírito Santo, Brazil. Revista Brasileira de Parasitologia Veterinária, 30, 1-5. doi:10.1590/S1984-29612021089
Wang, Q., Wang, H., Xie, M. 2010. Antibacterial mechanism of soybean isoflavone on staphylococcus aureus. Archives of Microbiology, 192, 89–898. doi:10.1007/s00203-010-0617-1

Download PDF

Version 1

posted

You are reading this latest preprint version

Effects of Mangifera indica leaf hay on weaned lambs before and after a Haemonchus contortus infection

Status:

Version 1

Abstract

1- Introduction

2- Material And Methods

2.1 Collection and preparation of hay from mango leaves

2.2 Animals And Diets Evaluated

2.3 Experimental Design

2.4 Evaluated Variables

Statistical analysis

3- Results

3.1 Animal performance

3.2 Blood Parameters And Fecal Nematode Egg Counting

4- Discussion

4.1- Animal performance

4.2 Conclusion

Declarations

References

Status:

Version 1