Hematological and serum biochemical profile of broilers during treatment with Vetom 21.77

 

Nozdrin G. A.¹, Rafikova E. R.², Yakovleva M. S.³

¹Professor, Novosibirsk State Agrarian University, Dobrolubova 160, Novosibirsk, 630039 Russia

²Research Lecturer, Novosibirsk State Agrarian University, Dobrolubova 160, Novosibirsk, 630039 Russia

Student, Novosibirsk State Agrarian University, Dobrolubova 160, Novosibirsk, 630039 Russia

 

ABSTRACT:

This study was carried out to investigate the effect of a new microbiological preparation Vetom 21.77 containing spore-mycelial biomass of Duddingtonia flagrans on hematological and biochemical parameters of broilers. The drug was administered for 3 months. A total number of 105 five-day-old broilers (Hubbard F15) were allocated to four test groups, where birds received the preparation in doses of 2, 5, 50 and 300 μl/kg bodyweight, and a control group with no treatment. Blood samples were collected 1.5 and 3 months after the beginning of the experiment for biochemical and hematological analysis. According to the results obtained, administration of the microbiological drug was followed by positive changes in the immunity of treated broilers. The preparation seems to have stimulating effect on their metabolic processes. There was no negative effect of the drug on the physiological state of the birds. In the course of the experiment, neither disease indications, nor mortality were detected in the birds, excluding deaths caused by the required slaughter. There were no treatment-related allergic reactions, even at the highest daily dose, as exemplified by almost borderline low eosinophil levels observed in treated broilers over the study period. No clinically relevant results were detected in serum biochemical profile.

 

 

 

INTRODUCTION:

The necessity of searching for safe and inexpensive compounds of non-chemical origin that have the potential to replace antibiotic growth promoters in poultry diets has repeatedly been pointed out by researchers worldwide [1-4].

 

An extensive list of natural growth stimulants has been accumulated in the literature to date. They are characterized by a beneficial effect on the state of the gastrointestinal tract. Sethiya has attempted to summarize these replacers for synthetic drugs [5].

 

 

 

 

Valenzuela-Grijalva et al. in their analytical review of relevant research on alternative growth promoters conclude that in the case of experiments on broilers those agents were effective in increasing their productivity, as well as meat quality [6].

 

A separate stream among organic antibiotic replacers is fungi. There is evidence from experiments with broilers and some animals for a positive impact on their performance resulting from dietary supplementation of Scytalidium acidophilum, Lentinus edodes, Ganoderma lucidum, Pleurotus ostreatus, Cordycepss inensis and Duddingtonia flagrans [7-11]. The latter is widely regarded as a bioagent antagonistically acting on various helminths. The newest studies also confirm its predatory capabilities [12,13]. The beneficial influence of this hyphomycete on animal digestive tract has inspired scientists to expand research into its additional abilities. Thus, for example, a new preparation Vetom 21.77 based on this carnivorous fungus was shown to increase live weight of mice [14].

 

A favorable impact of Vetom series preparations on hematological indices in broilers has previously been demonstrated by Nozdrin et al. [15], in particular. As distinct from other strains of Vetom, the one considered in the present study is based on the fungal spore-mycelial biomass which is supposed to normalize the physiological status of birds and animals by reducing the parasite burden. The preparation is at the preclinical trials stage and its influence on blood picture of birds is little known so far. Meanwhile, blood picture is an important criterion in assessing animal and bird health [16].

 

The aim of the current research was to determine the effect of different levels of Vetom 21.77, containing spore-mycelial biomass of D. flagrans, given to broilers on their hematological and serum biochemical parameters.

 

MATERIALS AND METHODS:

The study period extended for three months and was conducted with 105 healthy quarantined 5-day-old Hubbard F15 broilers of uniform weights. The birds were housed on concrete floors with wheat straw bedding, at the agrotechnopark of Shakarim State University (Semey, Kazakhstan). The conditions in the birds holding area were in accordance with Hubbard Broiler Management Manual [17]. This experiment was implemented as a part of a subchronic oral toxicity study in accordance with OECD Test No. 409 [18] in which Vetom 21.77 based on spore-mycelial biomass (1х109 CFUs/cm3) of D. flagrans F-882 was evaluated on mice and chickens.

 

The chickens were assigned into 4 treatment groups (received the preparation) and a control group (basal diet only), 20 birds per group. Five additional chickens were used for hematologic and biochemical analysis performed prior to the start of the experiment for the purpose of comparing with further results. Ingredient and nutrient composition of basic diets provided in pellet form were equal for broilers of all groups (Tab. 1). The preparation was given orally to broilers of 1th, 2nd, 3rd and 4th groups once-daily at 2, 5, 50 and 300 μl/kg body weight/day, respectively, daily over the period of 3 months. Every drug application was followed by visual observation of the birds.

 

 

Table 1. Composition of basal diets for broilers of a control and treatment groups

Diet features	Starter (0-14 days)	Grower (15-34 days)	Finisher (35 days and older)
Metabolizable energy (kcal/kg feed)	3150	3100	3180
Crude protein (%)	22.0	26.5	19.2
Raw fiber (%)	8.0	10.0	5.6
Raw fat (%)	3.47	8.8	7.3
Calcium (%)	2.5	4.8	1.1
Available phosphorus (%)	0.8	0.8	0.7
Lysine (%)	1.15	1.44	1.0
Methionine (%)	0.42	0.77	0.77

 

Blood samples were punctured from the wing vein of 5 broilers per group 1.5 and 3 months after the experiment had started, plus 5 chickens before. The procedure conformed to Bessarabov's guidelines [19]. The samples were collected in plain red-top, lavender-top and sure-sep vacutainer tubes, clotted and then centrifuged at 3000 rpm for 15 minutes to separate the blood into plasma, RBC and WBC layers. The samples were transferred to the secondary tubes, loaded onto the analyzers and processed in accordance with the manufacturers' instructions.

 

Hematological analysis was performed using an automated hematology analyzer PCE 90 vet (HTI, USA). The following hematological parameters were estimated: RBC count, WBC count, platelets, PCV, Hb concentration, basophils, eosinophils, neutrophils, lymphocytes, monocytes, reticulocytes and ESR. ESR was measured by a Sunostik Sclot S2 coagulation analyzer (Sunostik Medical Technology Co., Ltd, China). MCH and MCV were calculated.

 

Serum biochemical profile was determined by means of semi-automatic biochemical analyzers Minitecno (I.S.E S.r.l., Italy) and Stat Fax 3300 (Awareness Technology, USA). The serum biochemical parameters measured included: total protein, albumin, urea, total bilirubin, ALT, AST, α-amylase, glucose, phosphorus, calcium, potassium, sodium, magnesium, total cholesterol, creatinine.

 

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

 

Treatment differences between the reference and the test groups were compared using one-tailed Mann–Whitney U test in StatsDirect 3.1.14 (StatsDirect Ltd, UK). The level of significance was set at Р <0.05. Data are expressed as mean ± SEM.

 

RESULTS AND DISCUSSION:

As indicated in Tables 2 and 3, all the morphological and biochemical blood indices in the chickens were within normal physiological limits, which may demonstrate there were no pathologies in their organisms. It is in agreement with the results of the clinical examination.

 

1.5 months after daily administration of the drug to the broilers started, they shown values of the parameters indicating the respiratory function of the blood and the erythropoietic organs activity that exceeded those in the control group. Significant differences of 5.4% (P <0.05), 11.2% (P <0.001) and 17.6% (P <0.001) were found for the numbers of RBCs between the treated groups 2, 3, 4, respectively, and the control (Tab. 4). Nonetheless, all of the morphological indices obtained were still within the reference intervals. This data is consistent with previous observations in the trial of Hernández, where horses were provided with pellets containing spores of Mucor circinelloides and D. flagrans [20]. Chicks of the 4th group also surpassed the controls in the level of hemoglobin by 10.3% (P <0.05) and lymphocytes by 7.1% (P <0.05). Reticulocytes were not increased markedly, as well as platelets; consequently, the alterations in RBCs could not be associated with regenerative response to an anemic stimulus [21], although MCV was in the low normal range. Besides, we defined no inflammatory leukogram there. Necropsy examination revealed no evidence of hemorrhage.

 

Table 2. Hematological profile of Hubbard F15 broilers prior to the start of treatment with Vetom 21.77 (n = 5, x̄ ± µ)

Parameters	x̄ ± µ	Normal range
RBCs (1012/L)	2.61±0.03	2.5-3.5
WBCs (109/L)	24.14±0.63	20-40
Platelets (109/L)	52.36±2.02	32-100
PCV (%)	24.4±0.4	22-35
Hb (g/L)	88.72±1.46	70-130
МСН (pg)	3.,93±0.25	33-47
МСV (fL)	93.33±0.87	90-140
Reticulocytes (%)	0.46±0.02	0-0.6
ESR (mm/h)	2.54±0.02	2-3
Leukogram, %
Basophils	1.2±0.49	1-3
Eosinophils	7.6±0.75	6-10
Neutrophils	28.0±2.07	15-50
Lymphocytes	57.4±0.87	52-60
Monocytes	5.8±0.66	4-10

 

Table 3. Serum biochemical profile of Hubbard F15 broilers prior to the start of treatment with Vetom 21.77 (n = 5, x̄ ± µ)

Parameters	x̄ ± µ	Normal range
Total protein (g/L)	32.8±1.03	30-44
Albumin (g/L)	20.9±1.82	14-29
Urea (mmol/L)	1.03±0.19	0.26-2.04
Total bilirubin (mg/dl)	1.88±0.01	1.86-1.96
ALT (IU/L)	11.6±0.53	0.1-14.8
AST (IU/L)	174.0±9.9	125-269
α-amylase (IU/L)	613.0±21.54	299-840
Glucose (mmol/L)	15.7±1.31	7.14-19.1
Phosphorus (mmol/L)	1.82±0.2	1.22-3.9
Calcium (mmol/L)	3.89±0.14	3.75-6.75
Potassium (mmol/L)	5.24±0.09	4.9-6.4
Sodium (mmol/L)	18.84±0.31	17.5-20
Magnesium (mmol/L)	0.97±0.01	0.82-1.11
Total cholesterol (mmol/L)	4.73±0.24	2.3-5.5
Creatinine (mmol/L)	0.1±0.02	0.08-0.16

 

Table 4. Hematological profile of Hubbard F15 broilers treated and non-treated with Vetom 21.77, 1.5 months after the start of the study (n = 25, x̄ ± µ)

Parameters	Control	Treatment1	Treatment 2	Treatment 3	Treatment 4
RBCs (1012/L)	2.78±0.06	2.81±0.02	2.93±0.01*	3.09±0.02**	3.27±0.05**
WBCs (109/L)	30.11±2,5	30.05±1.22	30.54±1.22	31.13±1,37	32.28±2.52
Platelets (109/L)	85.98±3.27	78.92±8.12	86.24±5.92	83.14±4.49	88.92±3.58
PCV (%)	28.6±0.87	26.8±0.37	27.8±0.37	29.0±0.32	30.0±0.45
Hb (g/L)	100.72±3.48	100.8±2.05	102.08±0.42	105.26±1.25	111.12±1.3*
МСН (pg)	36.29±1.07	35.86±0.9	34.87±0.25	34.02±0.42	33.99±0.18
МСV (fL)	102.98±1.73	95.34±1.77	94.95±1.26	93.73±0.72	91.75±0.32
Reticulocytes (%)	0.21±0.03	0.24±0.04	0.23±0.01	0.25±0.04	0.25±0.03
ESR (mm/h)	2.66±0.05	2.68±0.12	2.72±0.09	2.78±0.02	2.8±0.04
Leukogram, %
Basophils	1.2±0.2	1.2±0.2	1.0±0	1.2±0.2	1.0±0
Eosinophils	7.0±0.63	7.0±0.32	7.4±0.51	6.8±0.49	7.0±0.63
Neutrophils	29.0±0.89	28.0±1.87	27.8±1.56	27.8±1.43	27.2±0.73
Lymphocytes	53.6±1.03	55.6±1.5	55.8±1.36	56.4±1.12	57.4±1.21*
Monocytes	9.2±0.2	8.2±0.73	8.0±0.63	7.8±0.37	7.4±0.51

Significant difference versus the control denoted by asterisk – Mann-Whitney test: * – P <0.05; ** – P <0.001.

 

Concerning the serum biochemical parameters, there was a significant difference (P <0.01) of 5.2% and 10.4% for total protein level in broilers which were administered 50 and 300 μl/kg of the drug, respectively, as compared with non-treated birds (Tab. 5). Considering that PCV was normal, along with calcium/phosphorus ratio, this plasma protein magnitude is not indicative of dehydration, and no physical signs of it were detected withal. AST values in these groups also outnumbered the control one by 6.93% (P <0.05) and 10.4% (P <0.05), respectively. All the values remained within the normal levels.

Table 5. Serum biochemical profile of Hubbard F15 broilers treated and non-treated with Vetom 21.77, 1.5 months after the start of the study (n = 25, x̄ ± µ)

Parameters	Control	Treatment1	Treatment 2	Treatment 3	Treatment 4
Total protein (g/L)	34.6±1.21	35.0±1.24	35.8±1.12	36.4±0.4**	38.2±0.44**
Albumin (g/L)	22.2±2.2	23.0±2.77	22.8±2.26	24.0±2.12	24.6±1.23
Urea (mmol/L)	1.36±0.04	1.33±0.02	1.4±0.13	1.39±0.03	1.42±0.03
Total bilirubin (mg/dl)	1.9±0	1.91±0.01	1.87±0.01	1.92±0.01	1.9±0
ALT (IU/L)	12.2±0.78	11.9±1.34	11.8±0.87	11.4±0.7	11.1±0.78
AST (IU/L)	202±5.46	213±10.01	210±4.30	216±2.19*	223±4.64*
α-amylase (IU/L)	590±22.65	611±24.17	615±15.73	596±22.69	602±17.05
Glucose (mmol/L)	12.46±0.84	12.31±0.77	12.03±1.11	12.19±0.90	12.05±1.11
Phosphorus (mmol/L)	2.02±0.23	2.06±0.16	2.09±0.17	2.05±0.17	2.11±0.18
Calcium (mmol/L)	4.01±0.09	3.94±0.09	4.0±0.05	3.99±0.11	4.04±0.05
Potassium (mmol/L)	5.94±0.21	6.04±0.12	6.1±0.12	5.98±0.17	6.02±0.14
Sodium (mmol/L)	19.04±0.31	19.0±0.28	18.92±0.19	19.02±0.21	18.98±0.14
Magnesium (mmol/L)	1.04±0.01	1.02±0.02	0.98±0.01	1.06±0.02	1.01±0.04
Total cholesterol (mmol/L)	3.16±0.33	3.11±0.03	3.19±0.24	3.15±0.22	3.1±0.02
Creatinine (mmol/L)	0.09±0	0.10±0.01	0.10±0.01	0.10±0.01	0.11±0.01

Significant difference versus the control denoted by asterisk – Mann-Whitney test: * – P <0.05; ** – P <0.01.

 

Table 6. Hematological profile of Hubbard F15 broilers treated and non-treated with Vetom 21.77, 3 months after the start of the study (n = 25, x̄ ± µ)

Parameters	Control	Treatment1	Treatment 2	Treatment 3	Treatment 4
RBCs (1012/L)	2.92±0.02	2.91±0.16	3.05±0.05	3.13±0.13	3.21±0.1*
WBCs (109/L)	30.86±2.63	31.41±2.90	32.54±2.71	32.92±2.27	33.27±1.31
Platelets (109/L)	77.06±3.04	80.66±5.98	79.42±7.25	80.12±6.34	78.04±3.36
PCV (%)	30.2±0.73	32.4±1.44	31.0±1.18	32.2±1.36	32.0±1.14
Hb (g/L)	105.2±1.69	103.2±4.07	106.5±2.52	107.0±3.99	112.7±3.08*
МСН (pg)	36.06±0.62	35.61±0.72	34.91±0.45	34.19±0.28	35.15±0.14
МСV (fL)	103.55±2.97	112.2±5.87	101.49±2.38	103.08±3.84	99.8±2.27
Reticulocytes (%)	0.16±0.01	0.13±0.03	0.16±0.03	0.15±0.03	0.15±0.03
ESR (mm/h)	2.72±0.07	2.74±0.07	2.9±0.03	2.84±0.09	2.92±0.04
Leukogram, %
Basophils	1.4±0.4	1.2±0.2	1.2±0.2	1.0±0	1.0±0
Eosinophils	6.8±0.37	7.2±0.49	7.0±0.32	6.8±0.58	6.6±0.4
Neutrophils	28.0±1.05	27.4±0.51	24.8±1.32	25.4±1.12	27.4±0.87
Lymphocytes	54.8±0.86	55.6±1.03	58.2±1.11*	58.8±0.73**	57.4±0.93*
Monocytes	9.0±0.32	8.6±0.4	8.8±0.37	8.0±0.32	7.6±0.24

Significant difference versus the control denoted by asterisk – Mann-Whitney test: * – P <0.05; ** – P <0.01.

 

Urea, creatinine and phosphorus were not elevated markedly; any glumeral filtration disorders were therefore unlikely to occur.

 

As reflected in Tab. 6, 3 months from the experiment beginning, alterations in the morphological parameters did not go beyond the corresponding reference values either.

 

Statistically significant outpace in relation to controls for RBC (9.9%, P <0.05) and Hb (7.1%, P <0.05) were observed in the 4th experimental group alone this time.

 

Silva et al. [22] argue that PCV concentrations in the blood of sheep that received pellets with AC001 isolate of D. flagrans were moderately lower in comparison with other groups. This trend was not confirmed in the current study.

 

It is important to note that during our experiment, MCH level, largely determining the intensity of oxidation-reduction processes in living organisms, did not exceed the physiological norm, although it was near the lower boundary of the reference interval in treatment 4.

At the same time, experimental groups 2, 3 and 4 showed lymphocytes counts higher than those in the control one: 6.2% (P <0.05), 7.3% (P <0.01) and 4.7% (P <0.05). Such dynamics may point to an increase in the adaptive immunity function intensity in the treated birds. In this respect, we note that according to the leukogram reflected in the work of Vilela et al. [23], there also was higher percentage of lymphocytes in the blood of goats fed pellets containing D. flagrans (AC001) in comparison with groups where this hyphomycete was not applicated.

 

As it can be seen in Tab. 7, the serum biochemical values did not fall beyond normal limits at this stage of the research. However, in general, the values in the test groups exceeded the control numbers to some extent.

 

 

Table 7. Serum biochemical profile of Hubbard F15 broilers treated and non-treated with Vetom 21.77, 3 months after the start of the study (n = 25, x̄ ± µ)

Parameters	Control	Treatment1	Treatment 2	Treatment 3	Treatment 4
Total protein (g/L)	36.2±0.55	36.8±1.99	37.6±0.61	38.0±0.27*	39.4±0.56*
Albumin (g/L)	24.0±2.63	24.2±1.8	24.6±2.07	25.0±2.15	26.2±1.65
Urea (mmol/L)	1.61±0.16	1.68±0.17	1.72±0.04	1.71±0.12	1.74±0.12
Total bilirubin (mg/dl)	1.92±0.01	1.9±0	1.9±0.01	1.88±0.01	1.89±0.01
ALT (IU/L)	14.1±1.37	13.7±0.5	13.4±0.35	13.4±0.69	13.2±0.38
AST (IU/L)	215.0±6.23	210.0±11.28	229.0±13.86	222.0±12.4	218.0±13.21
α-amylase (IU/L)	656.0±28.51	639.0±18.07	660.0±19.98	651.0±9.35	669.0±27.03
Glucose (mmol/L)	10.79±0.63	10.45±1.2	10.86±0.59	10.14±0.08	9.92±1.03
Phosphorus (mmol/L)	2.28±0.03	2.29±0.21	2.2±0.08	2.32±0.2	2.3±0.09
Calcium (mmol/L)	4.33±0.19	4.25±0.12	4.3±0.12	4.39±0.15	4.36±0.11
Potassium (mmol/L)	5.72±0.21	5.9±0.23	5.84±0.26	5.94±0.16	6.1±0.09
Sodium (mmol/L)	18.68±0.19	18.78±0.12	18.7±0.14	18.8±0.12	18.74±0.32
Magnesium (mmol/L)	0.96±0.02	0.93±0.01	0.95±0.01	1.01±0.04	0.95±0.03
Total cholesterol (mmol/L)	5.07±0.19	5.1±0.07	5.02±0.18	5.09±0.11	5.05±0.15
Creatinine (mmol/L)	0.08±0	0.09±0	0.09±0.01	0.09±0	0.09±0

Significant difference versus the control denoted by asterisk – Mann-Whitney test: * – P <0.01.

 

 

With regard to the total plasma protein level, treatment 3 had greater mean as opposed to the intact group (5%, P <0.01), as well as treatment 4 (8.8%, P <0,01). This is not in line with findings of Rocha et al [24]. The authors state a lack of significant changes in total amounts of serum protein between involved sheep caused by administration of pellets with D. flagrans.

 

When examining capacity of this predatory fungus to control Cyathostomins of horses, Buzatti et al. [25] found that the serum glucose was significantly different (P = 0.001) between two groups studied. No phenomenon akin to this was seen within the present survey.

 

In fact, given that the vast majority of studies about effects of D. flagrans upon the animal organism were focused on the gastrointestinal tract aspect, only serum pepsinogen concentrations were measured there among biochemical parameters of blood [26-28]. This makes it impossible for us to compare our results with those of other researchers completely.

 

CONCLUSION:

Thereby, Vetom 21.77, when applied at doses of 2, 5, 50 and 300 μl/kg body weight, in general, did not adversely affect physiological state, serum biochemistry or hematological parameters of the broilers.

 

The results suggest that the preparation had a positive influence on the physiological status of the birds through intensification of metabolic processes by increasing RBC and Hb counts in the normal range. It should be noted that MCH values displayed sufficient saturation of erythrocytes with Hb.

 

 

 

Test broilers had gradual shift of leukogram towards lymphocytes within the norm, which may indicate the preparation-related activation of cellular elements of immune system.

 

Neither disease indications, nor mortality were observed in the birds throughout the experiment, excluding deaths caused by the slaughter. There were no treatment-related allergic reactions, even at 300 μl/kg bodyweight dose, as evidenced by almost borderline low eosinophil levels observed in treated broilers over the study period. No clinically relevant results were registered in serum biochemical profile.

 

List of symbols and Abbreviations:

RBCs – Red Blood Cells

WBCs – White Blood Cells

PCV – Packed Cell Volume

Hb – Hemoglobin

ESR – Erythrocyte Sedimentation Rate

MCH – Mean Corpuscular Hemoglobin

MCV – Mean Corpuscular Volume

ALT – Alanine aminotransferase

AST – Aspartate aminotransferase

CFUs – Colony–forming Units

SEM – Standard Error of the mean

x̄ – the mean (average)

µ – Standard Error of the mean

 

ACKNOWLEDGMENT:

We thank Professor Abdrakhman Baigazanov for providing necessary facilities for the present work. Our special appreciation goes to Sergey Kozhevnikov who has translated the paper from Russian into English.

 

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

 

REFERENCES:

1.       Akinleye SB, Iyayi EA and Afolabi KD. The performance, haematology and carcass traits of broilers as affected by diets supplemented with or without biomin a natural growth promoter. World Journal of Agricultural Sciences. 4 (4); 2008: 467-470.

2.       Buchanan NP, Hott JM, Cutlip SE, Rack AL, Asamer A and Moritz JS. The effects of a natural antibiotic alternative and a natural growth promoter feed additive on broiler performance and carcass quality. Journal of Applied Poultry Research. 17 (2); 2008: 202-210.

3.       Huyghebaert G, Ducatelle R and Van Immerseel F. An update on alternatives to antimicrobial growth promoters for broilers. The Veterinary Journal. 187 (2); 2011: 182-188.

4.       Kulcsár A, Mátis G, Molnár A, Petrilla J, Wágner L, Fébel H and Neogrády Z. Nutritional modulation of intestinal drug-metabolizing cytochrome P450 by butyrate of different origin in chicken. Research In Veterinary Science. 113; 2017: 25-32.

5.       Sethiya NK. Review on natural growth promoters available for improving gut health of poultry: An alternative to antibiotic growth promoters. Asian Journal of Poultry Science. 10; 2016: 1-29.

6.       Valenzuela-Grijalva NV, Pinelli-Saavedra A, Muhlia-Almazan A, Domínguez-Díaz D and González-Ríos H. Dietary inclusion effects of phytochemicals as growth promoters in animal production. Journal of Animal Science and Technology. 59 (1); 2017: 8.

7.       Chandrawathani P, Jamnah O, Adnan M, Waller PJ, Larsen M and Gillespie AT. Field studies on the biological control of nematode parasites of sheep in the tropics, using the microfungus Duddingtonia flagrans. Veterinary Parasitology. 120 (3); 2004: 177-187.

8.       Huang MK, Choi YJ, Houde R, Lee JW, Lee B and Zhao X. Effects of Lactobacilli and an acidophilic fungus on the production performance and immune responses in broiler chickens. Poultry Science. 83 (5); 2004: 788-795.

9.       Gómez-Rincón C, Uriarte J and Valderrábano J. Effect of nematophagous fungus Duddingtonia flagrans and energy supplementation on the epidemiology of naturally infected kids. Veterinary research. 38 (1); 2007: 141-150.

10.     Jackson JN. Effects of Phase Feeding Different Levels of Fungus Myceliated Grain on Broiler Performance and Health (Doctoral dissertation, North Carolina Agricultural and Technical State University). Available from: URL:

10.http://cdm15116.contentdm.oclc.org/cdm/singleitem/collection/p9007coll2/id/643.

11.     Willis WL, Wall DC, Isikhuemhen OS, Jackson JN, Ibrahim S, Hurley SL and Anike F. Effect of level and type of mushroom on performance, blood parameters and natural coccidiosis infection in floor-reared broilers. The Open Mycology Journal. 7; 2013: 1-6.

12.     Rodrigues JVF, Braga FR, Campos AK, de Carvalho LM, Araujo J, Aguiar AR and de Freitas SG. Duddingtonia flagrans formulated in rice bran in the control of Oesophagostomum spp. intestinal parasite of swine. Available from: URL:

12.https://www.sciencedirect.com/science/article/pii/S0014489417303284.

13.     de Gives PM, Arellano MEL, Marcelino LA, Jenkins SO, Guerrero DR, Várgas GR and Murillo VEV. The nematophagous fungus Duddingtonia flagrans reduces the gastrointestinal parasitic nematode larvae population in faeces of orally treated calves maintained under tropical conditions. Dose/Response assessment. Available from: URL:

13.https://www.sciencedirect.com/science/article/pii/S0304401718300372.

14.     Rafikova ER and Nozdrin GA. A preliminary investigation on determination of a toxicity class of a new probiotic preparation vetom 21.77. Available from: URL: https://elibrary.ru/item.asp?id=30513565. (In Russian).

15.     Nozdrin GA. Regulations and characteristics of the action of probiotics of vetom class on blood morphological indexes in different animals. Available from: URL: https://elibrary.ru/item.asp?id=13616579. (In Russian).

16.     Etim NN, Enyenihi GE, Williams ME, Udo MD and Offiong EEA. Haematological parameters: indicators of the physiological status of farm animals. British Journal of Science. 10 (1); 2013: 33-45.

17.     Hubbard Broiler Management Manual, 2016. Available from: URL: 

17.https://www.hubbardbreeders.com/media/20160527__broiler_management_manual__fast_growth__gb_lh__068468600_1206_26102017.pdf

18.     OECD (1998), Test No. 409: Repeated Dose 90-Day Oral Toxicity Study in Non-Rodents, OECD Publishing, Paris. Available from: URL: http://dx.doi.org/10.1787/9789264070721-en.

19.     Bessarabov BF. Laboratornaja diagnostika klinicheskogo i immunobiologicheskogo statusa u sel'skohozjajstvennoj pticy. KolosS, Moscow. 2008. (In Russian).

20.     Hernández JÁ, Arroyo FL, Suárez J, Cazapal-Monteiro CF, Romasanta Á, López-Arellano ME and de Gives PM. Feeding horses with industrially manufactured pellets with fungal spores to promote nematode integrated control. Veterinary Parasitology. 229; 2016: 37-44.

21.     Thrall MA, Weiser G, Allison R and Campbell T. Veterinary hematology and clinical chemistry. John Wiley & Sons, Ames. 2012.

22.     Silva BF, Carrijo-Mauad JR, Braga FR, Campos AK, Araújo JV and Amarante AF. Efficacy of Duddingtonia flagrans and Arthrobotrys robusta in controlling sheep parasitic gastroenteritis. Parasitology Research. 106 (6); 2010: 1343-1350.

23.     Vilela VLR, Feitosa TF, Braga FR, de Araújo JV, de Oliveira Souto DV, da Silva Santos HE and Athayde ACR. Biological control of goat gastrointestinal helminthiasis by Duddingtonia flagrans in a semi-arid region of the northeastern Brazil. Veterinary Parasitology. 188 (1-2); 2012: 127-133.

24.     Rocha RA, Araújo JV and Amarante AFT. Efficacy of the nematode-trapping fungus Duddingtonia flagrans against infections by Haemonchus and Trichostrongylus species in lambs at pasture. Journal of Helminthology. 81 (4); 2007: 387-392.

25.     Buzatti A, Santos CP, Yoshitani UY, Sprenger LK, Kloster F, Antunes JD and Molento MB. Biological control using the fungi Duddingtonia flagrans against cyathostomins of horses. Journal of Equine Veterinary Science. 32 (10); 2012: S31.

26.     Fässler H, Torgerson PR and Hertzberg H. Failure of Duddingtonia flagrans to reduce gastrointestinal nematode infections in dairy ewes. Veterinary Parasitology. 147 (1-2); 2007: 96-102.

27.     Paraud C, Pors I and Chartier C. Efficiency of feeding Duddingtonia flagrans chlamydospores to control nematode parasites of first-season grazing goats in France. Veterinary Research Communications. 31 (3); 2007: 305.

28.     Epe C, Holst C, Koopmann R, Schnieder T, Larsen M and von Samson-Himmelstjerna G. Experiences with Duddingtonia flagrans administration to parasitized small ruminants. Veterinary Parasitology. 159 (1); 2009: 86-90.

 

 

 

 

Accepted on 10.07.2018          © RJPT All right reserved

Research J. Pharm. and Tech 2019; 12(8): 3739-3744.