Induced Ruminal Lactic Acidosis in Sheep Treated with Various Remedial Agents in Libo Kemkem Districts, Northwest Ethiopia

Background Grain overload is a ruminant metabolic disorder associated with overingestion or a sudden change to rapidly fermentable concentrate feeds. Objective A randomized clinical trial was carried out in Libo Kemkem districts to investigate vital signs, ruminal fluid, and hematological changes in sheep related to grain overload and to assess the treatment efficacy of various remedial agents in Farta sheep breeds. Methods All sheep were selected by using the simple random process, and categorized into four groups of eight animals groups I, II, and III in which each sheep was given 80 g of wheat flour per kg of body weight then baking soda, Magnalax, and digestive powder were treated successively, but group IV was the negative control group. Results The current clinical trial explained that all acidotic sheep had statistically significant (p < 0.0002) lower body temperature, rumen motility, protozoa activity, and ruminal fluid pH with 38.48 ± 0.20, 0.5 ± 0.89, 6 (75%), and 5.37 ± 0.34 mean value successively. Nevertheless, there were significant (p < 0.0059) increments in hematological variables including total red blood cell, total white blood cell, packed cell volume, and hemoglobin estimation with 14.05 ± 1.27, 12.71 ± 0.99, 40.78 ± 5.24, and 14.78 ± 1.83 mean value, respectively, before treatment in acidotic sheep. There were also vital sign increments including pulse rate, respiratory rate, skin turgor, and capillary refill time with 123.5 ± 27.1, 73.75 ± 12.71, 3 ± 1.78, and 3.37 ± 1.77 mean values, respectively, before medical treatment in acidotic sheep. Digestive powder was the first drug of choice, followed by Magnalax, and baking soda was ranked at the end based on clinical recovery. Conclusion The treatment effectiveness illustrated that all treatments could cure the grain overload. Nevertheless, digestive powder is the drug of choice based on the clinical and systemic recovery of the sheep. In addition to this, sheep should be introduced gradually to concentrate rations over 2–3 weeks with a mixture of concentrate in the milled feed containing roughage.


Introduction
Ruminal lactic acidosis is a metabolic disorder defned by decreased blood pH and bicarbonate, caused by overproduction of ruminal D-lactate [1].It results from the high production of lactic acid in the rumen after ingestion of large quantities of readily digestible carbohydrates, particularly grain.Wheat, barley, and corn are the most readily digestible grains; oats are less digestible [2].Rumen overload in sheep occurs due to the ingestion of grains, which helps the development of Streptococcus bovis bacteria [3].Morbidity rate and mortality rate vary based on the type of grain, the dose of grain eaten, and body condition score [2].
Te clinical signs of grain overload include reduction of appetite, fuid and electrolyte loss due to diarrhea, ataxia, grinding of teeth, grunting, and abdominal distension [4].Te treatment of ruminal acidosis depends on the degree of severity of the case.A variety of treatments can be given like antibiotics to reduce the growth of Gram-positive bacteria and fuid and electrolytes to maintain circulating blood volume [5].Sodium bicarbonate, magnesium hydroxide, and digestive powder are remedial agents that correct the disturbed blood pH to a normal level.Digestive powder is an appetizer, which induces reticulorumen motility.It consists of diferent components: paracetamol, glucose monohydrate, sodium propionate, and sodium hydrogen carbonate.Sodium chloride oral solution is given to treat fuid and electrolyte loss due to diarrhea [6].Sodium bicarbonate or magnesium hydroxide is applied to alkalinize the rumen pH (power of hydration).Fiber supplementation is given for the prevention of ruminal acidosis.Sheep should be introduced gradually to concentrate rations over 2-3 weeks, beginning with a mixture of ≤50% concentrate in the milled feed containing roughage [7].
Sheep and goat production in Ethiopia contributes a main role to the GDP (gross domestic product) of the country [8].Nevertheless, grain overload has a signifcant economic impact by resulting in delayed marketing, condemnation of entire carcasses, reduced weight gain, reduction of appetite, and death of sheep [9].
Various remedial agents have diferent scientifc names but are used for the correction of disturbed parameters related to ruminal lactic acidosis.It includes magnesium hydroxide or Magnalax, sodium bicarbonate or baking soda, and digestive powder, which are used to correct the disturbed ruminal and systemic acidosis to the normal required level of pH value in animals.Terefore, they have similar characteristics as the alkalinizer of ruminal acidosis [10].However, diferentiation among remedial agents for the treatment of ruminal and systemic acidosis due to gain overload is essential for accurate and rapid therapy of this disease.Tere has been no study about acute carbohydrate engorgement in sheep and its treatment efcacy of various remedial agents in Farta sheep breeds in Libo Kemkem districts, North West Ethiopia Terefore, the objective of this study is to investigate the treatment efcacy of various remedial agents of induced carbohydrate engorgement in Farta sheep breeds.

Study Area.
Te study was carried out in the Libo Kemkem district.It is placed in the south Gondar zone of the Amhara regional state (Figure 1).Te district has a latitude and longitude of 12 °07′N 37 °47′E and an elevation of 1975 meters above sea level.Te animal population consists of 124,607 cattle, 157,811 small ruminants, 27,329 equines, 199,950 poultry, and 21,381 honeybee hives [11].

Study Design.
A randomized clinical trial was carried out in Libo Kemkem districts.

Study Population.
Indigenous sheep breeds (Farta sheep) were brought from the market with a variety of ages and sexes selected in the Libo Kemkem district.

Study Animals.
A total of 32 Farta sheep breeds were selected by using the simple random method with a mean body weight of 28 kg (range 25-34 kg).Te sheep were given straw before and during the clinical trial.Sheep was of feed for twenty-four hours before the start of the clinical trial.During the clinical trial, each group was penned separately in their room.For this experiment, 32 sheep were randomly divided into 4 groups of eight animals in each group.On day zero, all sheep were ofered 80 g of wheat four per kg of body weight.Te four was ofered as a suspension in warm water by using a stomach tube [12].

Study Methodology
2.5.1.General Examination.All groups were observed three times in day associated with their posture, appetite, rumination, and diarrhea.General examination with clinical signs of anorexia, apathy, grinding of teeth, grunting, abdominal distension, ruminal stasis, watery diarrhea, ataxia, and recumbency were observed after the grain was ofered to sheep.Systemic examinations were carried out before and after treatment [6,11].
(1) Pulse and Respiration Taking.Te pulse rate was taken at the femoral artery [13].Te fngers were put on the femoral artery and gentle pressure was applied until the wave was determined.Te respiration rate was counted by numbering the movements of the rib cage and abdomen [14].
(2) Temperature Taking.Te body temperature was measured using a digital clinical thermometer.Te digital thermometer was gently inserted into the rectum.It was kept in the rectum for two minutes [15,16].

Observation of Ruminal Fluid
(1) Ruminal Fluid pH.Rumenocentesis was done to collect ruminal fuid by using 16 gauge needles with a disposable syringe.Te penetration site was the ventral rumen on the left side fve cm posterior to the last rib.Te procedure was done based on the standard [17].Te ruminal fuid was used for the evaluation of ruminal pH, which was measured using pH indicator paper or pH meter [18].Te ruminal fuid was placed on the paper and pH was observed by changing the color of the litmus paper and diferentiating it from the standard colors of the indicator paper.Te ruminal fuid was placed in a beaker and then a pH meter was inserted into it.So, the pH value was evaluated as the mean value [19].

Assessing of Hematological Parameter
(1).Blood Sample Collection.Blood sample collection was sampled from the jugular vein containing fve ml with tubes without anticoagulant and tubes containing EDTA (ethylene 2 Veterinary Medicine International diamine tetra acetic acid).Total white blood cell count (TWBC), total red blood cell (TRBC), hemoglobin estimation (Hb %), and packed cell volume (PCV) were evaluated by using an automatic haem-analyzer.Te hematological evaluation was done through an automated hematology analyzer [16].
(2) Determination of Blood pH.A blood sample was taken from the jugular vein by using a test tube containing 5 ml.
Te sample was permitted to clot at room temperature for one hour to obtain serum [17].Te serum pH was evaluated by using pH indicator paper and pH meter [20].Te pH indicator paper was inserted into serum and the color of the strip was diferentiated from the standard colors [18].Te serum was put into a beaker and a pH meter was inserted into it.Te mean value of both readings was evaluated [21].

Clinical Trials.
Clinical trials were carried out to diferentiate the best remedial agents.Bicart-sodium bicarbonate, digestive powder, and Magnalax-magnesium hydroxide treatment options were used for ruminal lactic acidosis as indicated in Table 1.Te remedial agents were ofered randomly to each group.Group I animals were given baking soda at a dose of 1 g/kg body weight.Group II was given Magnalax-magnesium hydroxide at a dose of 1.11 g/kg body weight.Animals in group III were treated with digestive powder at a dose of 0.8 g/kg.Te remaining sheep were considered as negative control (group IV).Sampling was performed before the onset of therapy and thereafter on post-treatment [21,23].Te therapeutic efectiveness was evaluated based on the disappearance of clinical signs, change of disturbed vital signs, ruminal pH, and hematological variable to the normal range [24].It was also evaluated by the survival rate.Finally, the accuracy of the remedial agent was assured based on the clinical recovery of the sheep and rumen function tests [23].
2.6.Data Analysis.Data were collected and entered into an MS Excel spreadsheet.Ten, it was processed using Stata MP 17. Te data were analyzed using ANOVA and t-test.Te hematological variable, ruminal, and clinical variables before and after therapy were calculated by one-way ANOVA.Te acidotic groups were diferentiated before and after therapy from the negative control group by paired t-tests.P value <0.05 was considered as statistically signifcant.3).Te rumen pH mean values observed after the remedial agent were 6.31 ± 0.43, 6.58 ± 0.30, 6.42 ± 0.37, and 6.65 ± 0.25 for groups I, II, III, and IV successively.Te increase in ruminal fuid pH value after remedial agent in all groups of acidotic sheep was statistically signifcant (p < 0.05) as assessed to their pretreatment value.

Protozoa Activity.
Ruminal protozoa want appropriate pH for their survival.Tere was a signifcant (p < 0.05) reduction in protozoa motility of acidotic sheep as we diferentiated to negative control.After the remedial agent, the protozoa activity test was carried out for all groups.Te protozoa motility in group II was the lowest as we contrasted to groups I, III, and IV as summarized in Table 3. Te improvement in protozoa motility after medical treatment in all the groups of acidotic sheep was statistically signifcant (p < 0.05) as contrasted to their pretreatment value.4.

Remedial Accuracy Determination.
Treatment accuracy of the remedial agents was assured based on clinical recovery from systemic acidosis.Te summary of treatment efcacy is illustrated in Table 5.When we compared the remedial agent efcacy, digestive powder was the frst drug followed by Magnalax and baking soda was ranked at the end based on the clinical recovery of the sheep.

Discussion
4.1.Vital Sign Parameter.Te reduction of body temperature observed in the current study had similar fndings to [25], which was due to fuid and electrolyte loss [4].Te increment of pulse rate was due to metabolic acidosis activation of the sympathetic nervous system [26,27].Te increased respiration rate noted in the current study had similar fndings to [28,29].Te elevation of respiration rate above normal level was due to activation of medulla oblongata because of increased carbon-dioxide tension of blood and reduced blood pH [27,30].

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Te occurrence of hypomotility or muscle atony during grain overload was due to the deactivation of the sympathomimetic ganglion [31].Te elevation of CRT and skin turgor in acidotic sheep had similar fndings [32,33].Tis is due to profuse diarrhea [27].

Ruminal Fluid Examination.
Te reduction in pH of the ruminal fuid was observed in the current study due to the increasing production of volatile fatty acids.Ingestion of fermentable carbohydrates leads to a change in the microfora in the rumen within 2-6 hours [29].Te Streptococcus bovis increases in number rapidly, which leads to the production high amount of lactic acid.When the rumen pH is below fve, it kills the microfora of the rumen.Lactobacilli bacteria ferment again the carbohydrate and induce a high amount of lactic acid [33,34].Te superimposition of lactic acid and lactate in the rumen liquid results in the movement of fuid into the rumen and so, leads to dehydration.Te present study had similar fndings with [11,18,25].
Te activity of the ruminal protozoa depends on the H + ion and osmolality of the rumen liquor.In the present study, ruminal protozoa motility in acidosis sheep was usually absent which had a similar report from [35].Magnalax treatment increased the rumen pH, but it reduced rumen protozoa motility.Tis fnding was also reported by [30,36,37].

Hematological Parameters.
Te elevation of TRBC, TWBC, PCV, and Hgb in acidotic sheep was observed similar to [36,38].Tis is due to fuid and electrolyte loss because of profuse diarrhea [39].Te increment in TRBC in lactic acidotic sheep was due to the stress efect of systemic acidosis and epinephrine releasement causing splenic contraction [40,41].Te current fndings of increased TRBC in ruminal acidotic sheep are in agreement with [37].
Te elevation of TWBC in ruminal acidosis is due to the production of endotoxins [34,42].Tis fnding was also reported by [38].
Elevation of PCV in systemic acidosis sheep was due to hem concentration and dehydration, which was also reported by [41].
Te elevation of hemoglobin concentration in the current study in lactic acidotic sheep had similar fndings with [4,30,40].Te clinical recovery and systemic changes after treatment have occurred, which were also reported by [32,43].
Te blood pH of ruminal lactic acidotic sheep in the present study was below the normal value, which was due to lactate absorption from the rumen.Tis report had similarities with [44].Te elevation of blood pH after medicinal treatment was due to the bufering or alkalinizing efect of the remedial agent as in agreement with [42,45].

Remedial Efcacy Assessment.
In the current study, baking soda, Magnalax, and digestive powder were applied as remedies for ruminal lactic acidosis.All remedial agents were able to treat the acidotic condition as described in Table 5.When we compared the remedial agent efcacy, digestive powder was the frst drug, followed by Magnalax and baking soda was ranked at the end based on the clinical recovery of the sheep.Tis showed that digestive powder was the most efective remedial agent which is in close agreement with [39,45].

Conclusion and Recommendations
Ruminal lactic acidosis is a metabolic disorder defned by decreased blood pH and bicarbonate, caused by overproduction of ruminal D-lactate.Te present clinical trial indicated that the acidosis sheep had lower body temperature, rumen motility, protozoa activity, and ruminal fuid pH.However, there was an elevation in pulse rate, respiration rate, CRT, and skin turgor.Tere were also signifcant (p < 0.05) increments in TEC, TLC, PCV, and Hgb in acidosis sheep.After the remedial agent application in acidotic sheep, the clinical and hematological parameters returned to normal levels.Te current fnding showed that all remedial agents were able to treat the acidotic condition.Digestive powder has various compositions, which are absorbed into the body and correct the disturbed clinical and hematological parameters.Digestive powder was the drug of choice, followed by Magnalax and baking soda was ranked at the end based on clinical and systemic recovery.In addition to this, sheep should be introduced gradually to concentrate rations over 2-3 weeks with a mixture of concentrate in the milled feed containing roughage.
Based on the above conclusions the following recommendations are forwarded: (i) We shall avoid sudden feed change from roughage to concentrate to reduce the occurrence of ruminal lactic acidosis (ii) Digestive powder has various compositions and can correct hematological and clinical changes easily and rapidly (iii) Additional studies had better be performed about the cause of ruminal lactic acidosis in ruminants

Table 2 .
Te mean rectal temperature values recorded before treatment were 38.48 C °± 0.20, 38.6 C °± 0.21, 38.71 C °± 0.54, and 39.32 C °± 0.27 for groups I, II, III, and IV, respectively, as indicated in Table 2. Te mean rectal temperatures calculated after treatment were 39.02 C °± 0.32, 39.11 C °± 0.14, 39.23 C °± 0.33, and 39.32 C °± 0.27 for groups I, II, III, and IV, respectively.Te mean pulse rate values before treatment for groups I, II, III, and IV were 123.5/min ±27.15, 115.5/min ±18.89, 117.25/min ±21.29 and 82/min ±6.19, respectively, as shown in Table 2.After treatment, the pulse rate of the groups I, II, III, and IV were again determined.Te mean pulse rate values calculated after treatment were 93.5/min ±30.61, 88/min ±6.68, 86/min ±7.37, and 82/min ±6.19 for groups I, II, III, and IV, respectively.Te mean skin turgor values counted before treatment were 3.37 ± 1.77, 4.12 ± 2.43, 3.12 ± 1.88 and 1 ± 0.00 second for groups I, II, III, and IV, respectively, as shown in Table 2. Tere was a signifcant (p < 0.05) increment in skin turgor of all groups of acidotic sheep as compared to healthy control.Te mean skin turgor values of

Table 4
III, and IV after treatment was 12.71 ± 1.99, 11.8 ± 0.92, 11.5 ± 0.85, and 11.32 ± 0.54, respectively, as described in Table4.Te total leukocyte count of ruminal acidotic sheep of groups I, II, and III was evident from values of 12.71 ± 0.99, 12.3 ± 1.82, and 11.83 ± 1.12, respectively, over 9.05 ± 0.80 of group IV before treatment.Te increment in total leukocyte count in all three ruminal acidotic groups was highly signifcant (p < 0.05) as compared to the healthy control group.Te total leukocyte count declined to 10.95 ± 1.73, 10.51 ± 1.20, 10.3 ± 0.89, and 9.05 ± 0.80 in groups I, II, III, and IV, respectively, after therapy in acidotic sheep as shown in Table4.Te average hematocrit (PCV) in ruminal acidotic sheep of groups I, II, and III were 40.78 ± 5.24, 40.5 ± 5.93, and 39.28 ± 20.86 over 29.41 ± 3.25 of group IV before treatment.Te hematocrit values declined to 35.27 ± 6.24, 33.87 ± 3.78, 31.68 ± 2.71, and 29.41 ± 3.25 in groups I, II, III, and IV, respectively, after treatment as indicated in Table4.Te blood pH of all ruminal acidotic sheep before treatment was found to be lower than the blood pH of the healthy control group.Te mean blood pH values recorded before treatment were 6.91 ± 0.44, 6.92 ± 0.24, 7.02 ± 0.24, and 7.26 ± 0.17 for groups I, II, III, and IV, respectively.Te blood pH values obtained after treatment were 7.11 ± 0.28, 7.31 ± 0.18, 7.21 ± 0.20, and 7.26 ± 0.17 for groups I, II, III, and IV, respectively, as shown in Table

Table 2 :
Clinical variables for a variety of groups due to grain overload efect (values are mean ± SE).Superscripts indicate statistical signifcance at p < 0.05 as it diferentiated to group IV. *

Table 3 :
Ruminal fuid analysis was performed in indigenous sheep breed to check protozoal activity (values are mean ± SE).Superscripts describe statistical signifcance at p < 0.05 as it contrasted to group IV. *

Table 4 :
Hematological variables change in indigenous sheep breed due to grain overload (values are mean ± SE).
* Superscripts indicate statistical signifcance at p < 0.05 as it contrasted to group IV.

Table 5 :
Remedial efectiveness of various therapeutic regimens in ruminal lactic acidosis in sheep.