Haematological Traits of Induced Hypovolemic Large White Yorkshire Piglets as Affected by Feeding them Milk of Pantja Goats and Black Badri Cows

Milk from goats and cows is invariably consumed by human beings throughout the globe. Goat milk has been said to be beneficial due to its unique nutritional and medicinal values (Holmes et al., 1946; Sharma and Kumar, 2004) and many times it has been prescribed as perfect alternative to cow and human milk, particularly for infants and aged people (Devendra and Burns, 1970, Chandan et al., 1992, Park and Haenlein, 2007). Goat milk is enriched with selenium and also improves digestive and metabolic utilization of various minerals (Gunjan et al., 2011). Cow milk is a rich, cheap and easy source of protein and calcium. It has important growth supporting minerals (K, Mg, P, Zn), which are absorbed in gut due to high lactose content. Further, favourable role of indigenous cow milk has been frequently mentioned in ancient and modern literature (Kalyan, 1945 and 1995). Old generation people have signified role of black Indian cow’s milk for maintaining better health and this notion is still prevalent in Indian society (Kalyan, 1945a). In India, dengue fever has been a common health issue during mosquito breeding season where-in there becomes a complication of selenium deficiency and reduced blood platelet count. Most of the people prefer goat milk during dengue fever and they give it upper hand for recovery without having any scientific proof.
       
Other than laboratory rodents, pig is recognized as an advantageous non-rodent animal model within a large number of biomedical research areas (Roth and Tuggle, 2015). Considering the above facts present work was undertaken to study effect of feeding milk of Pantja goats and black Badri cows to experimentally induced hypovolemic large white yorkshire (LWY) weaned piglets to ascertain superiority of goat milk over cow milk or vice-versa, that may be recommended for improving haematological traits during the conditions as prevailing during dengue fever, accidental hypovolemia and the like.

Before conducting the experiment, an approval (IAEC-CVASc-LPM-385) was obtained from duly constituted Institutional Animal Ethics Committee. The experiment was conducted at Pig unit of LPM Department of College of Veterinary and Animal Sciences, GBPUAT, Pantnagar during October -November 2019 and May-June 2020. The place is situated in Tarai plains at the foothills of Shivalik ranges of Himalayas at 29.5^oN and 79.5^oE at 243.84 m MSL, experiencing ambient temperature exceeding 42^oC in summer and below 2oC in winter and relative humidity range of 15 to 95 per cent.
       
A total of 18 LWY weaned piglets, aged 2.5 months, littered by 3 sows were available for the study. They were individually sexed and weighed and divided into three groups to conduct the experiment in randomized block design for a period of 30 days. All these groups had 6 piglets in each (having 4 males and 2 females in each group), with an average body weight of 8.91± 3.33, 8.78 ± 2.09 and 9.00 ± 2.42 kg for T1, T2 and T3 groups, respectively. All the piglets were reared under intensive system in well ventilated pens on concrete flooring and provided with similar environmental and managemental conditions. 
       
In order to attempt for induced hypovolemia, first total volume of blood in circulation in the piglet was estimated as per IACUC (2007), followed by taking out 15% of the piglet’s total estimated blood volume [@ 7.5% on day 1^st (75^th day age) and again 7.5% on day 3^rd (78^th day age)], as suggested by IACUC (2007). All the 18 piglets of the three groups were subjected to induced hypovolemia by withdrawing required amount of blood at a fixed schedule (Table 2; Dineshkumar, 2020).
 
Feeding schedule of induced hypovolemic LWY piglets
 
The control group piglets (T1) were maintained on basal diet only during the experimental period. Whereas, the treatment groups piglets, T2 and T3, were provided with freshly collected milk of farm reared Pantja goats and black Badri cows (Table 1), respectively, in addition to the usual basal diet for a period of 30 days (i.e. from 75^th to 135^th day age of piglets). The milk was hand fed through pail and its quantity was double the volume of blood, withdrawn from the piglets to induce hypovolemia (Table 2). Ad libitum drinking water was made available to all piglets throughout the period of experiment.
 

 

       
Sampling of blood (5 ml) from each piglet was done on day 1^st, 3^rd, 7^th, 15^th and 30^th of the experiment (i.e. 75^th, 78^th, 82^nd, 90th and 105^th day age of piglets) using a sterilized disposable syringe and blood was immediately transferred to EDTA rubber capped 10 ml tube. On first two occasions, the samples were taken from the removed blood while attempting for induced hypovolemia. However, the blood for the remaining three samples was collected by following the usual procedure. All the 90 samples, so collected, were kept at room temperature in thermos and analysed through Auto-haem Analyser (Celltac-α, MEK-6510K) on the same day of their collection. The values so obtained for haemoglobin (g/dl), erythrocyte sedimentation rate (mm/h) and blood platelets count (x10⁵ cells/ mm³) were subjected to statistical analysis as per Snedecor and Cochran (1994).

Several physical signs to indicate hypovolemia [postural vital signs such as increase in heart rate by >30/min, decrease in systolic blood pressure by >20 mmHg, dryness of mouth, nasal mucous membranes, axillae and tongue and neurological signs like alterations in mental status, weakness of upper or lower extremities] have been reported (Gergely et al., 2011). However, as a scope of present study following paragraphs discuss changes in haemoglobin, ESR and blood platelets counts of experimentally induced hypovolemic weaned LWY piglets as affected by feeding them with milk of Pantja goats and black Badri cows.
 
Haemoglobin
 
The overall value for haemoglobin in LWY piglets was 10.85±0.11 g/dl. The values for control, Pantja goat milk fed and black Badri cow milk fed group of piglets for haemoglobin were 10.66±0.27, 10.95±0.16 and 10.93±0.17 g/dl, respectively, which did not vary significantly. However, the values showed significant (P<0.01) variation on different days following hypovolemia, being higher on 1^st and 30^th day and the lowest and insignificantly different on 3^rd, 7^th and 15^th day. Overall haemoglobin value on 30^th day became similar to normal value, 11.36 ± 0.20 g/ dl and was in consonance with reported values (Reece et al., 2015). About 8-12% of initial haemoglobin level was reduced due to blood withdrawal. These low values may be a consequence to experimental hypovolemia till 15^th day (Table 3, Fig 1).
 

 

       
The values for haemoglobin for control, Pantja goat milk fed and black Badri cow milk fed groups piglets did not show any significant variation on various days of hypovolemia, implying that all the group piglets behaved similarly in declining and then regaining the haemoglobin level over the study period. The overall values were numerically, but not statistically, highest in the order of “Pantja milk fed > black Badri cow milk fed > Control group” (Table 3).
       
Haemoglobin is a complex iron containing and oxygen carrying pigment conjugated with protein in red blood cells. Its low amount leads to anaemia, causing depletion of oxygen in tissues and overall inactivity and other related problems. Normal haemoglobin range in pig is reported as 10.0-16.0 g/dl (Reece et al., 2015).
 
Erythrocyte sedimentation rate (ESR)
 
Overall values for erythrocyte sedimentation rate was 7.87± 0.74 mm/hr. Control, Pantja goat milk fed and black Badri cow milk fed groups had ESR values of 7.30±1.16, 9.70±1.77 and 6.60±0.98 mm/hr, respectively and they didn’t show any significant variation among the groups. However, on different days following hypovolemia these values had a significant variation (P<0.05), being higher on day 1^st and 3^rd and lower on day 7^th, 15^th and 30^th (Table 4, Fig 1). These low values may be due to simultaneous increase in total erythrocytic count which in turn increases viscosity of blood (Dineshkumar, 2020). Reduction in ESR value indirectly indicates improvement and thus was a good sign for healthy animal.
 

       
Control, Pantja goat milk fed and black Badri cow milk fed groups piglets’ ESR values on different days of hypovolemia did not show any significant variation, implying that all the treatment groups behaved similarly during the study period (Table 4).
       
ESR in an anticoagulated whole blood is the rate at which red blood cells descend in a vertically placed standard tube over a period of time. Generally, it is measured in millimetre per hour (mm/ h) as the distance in uppermost layer of erythrocyte descends in certain length. It is mainly based on degree of agglutination of erythrocyte and level of plasma proteins (fibrinogen, albumin, globulin, etc.). In swine, ESR normally ranges from 1-14 mm/hr (Reece et al., 2015).
 
Platelets count (x105cell/ mm3)
 
Platelets are the tiny blood cells that help to stop bleeding by forming clots. The process of spreading across the surface of damaged blood vessels to stop bleeding is adhesion by mash work with fibrin. Platelets count in domestic animals range from 200,000-400,000 cells/mm³. Reduction in platelets count is termed as thrombocytopenia, which may lead to haemorrhage in internal organs (Reece et al., 2015).
       
The overall mean platelets count (x105 cells/ mm3) for LWY piglets was 3.257± 0.089. The values of platelets count in control, Pantja goat milk fed and black Badri cow milk fed groups piglets were 3.0983±0.1675, 3.0820±0.1490 and 3.5907±0.1885, respectively, being significantly higher (P<0.05) for black Badri cow milk fed group piglets. Whereas, the values for platelets count were statistically indifferent for control and Pantja goat milk fed group piglets (Table 5, Fig 1). This implies that use of indigenous cow milk (black Badri cow milk), but not the goat (Pantja) milk, may be helpful in improving platelets count in human beings too, as pig is a good model for human medical studies. However, further studies are required to validate this finding. Jung et al., (2008) observed the platelets (x 105cell/mm³) as 13.74, 6.55, 4.53, 4.77 on 4, 8, 12 and 20 weeks of age in miniature growing pigs.
 

       
The platelets count (x10⁵ cell/ mm³) during different days of testing were 3.1767±0.2577, 2.7606±0.2022, 3.9717± 0.2045, 3.3172±0.1592 and 3.0589±0.2057 on day 1^st, 3^rd, 7^th, 15^th and 30^th, respectively. These values were significantly higher (P<0.01) on day 1^st, 7^th, 15^th and 30^th day, but not on 3^rd day of experiment. Initially, on 1^st day platelets count was in normal range and after blood withdrawal there was 11-13% reduction in cells from the initial level. Its low value on day 3^rd of testing was a sequel to induced hypovolemia. Level of platelets counts became higher on 7^th day and later normal in all the treatment groups piglets. However, platelets count was significantly higher (P<0.05) in black Badri cow milk fed piglets than the rest groups, indicating positive role of black Badri cow’s milk feeding on improving platelets in hypovolemic piglets (Table 5). Since pig is a good model for human medical researches, present findings could be beneficial for human beings also for improving blood platelets counts in Dengue fever and like situations where thrombocytopenia becomes the major health issue.
       
Other than laboratory rodents, pig is recognized as an advantageous non-rodent animal model within a large number of biomedical research areas (Roth and Tuggle, 2015) and it is a leading animal species within studies of xenotransplantation of animal organs into humans (Schook et al., 2005). So far there seems no similar study that has been carried out with regard to using goat milk, indigenous cow milk or indigenous black cow milk to increase count of blood platelets or other blood components in vivo. At present such nature of studies are rare and the proofs for review or validation were scanty in literature, so further studies are recommended on these lines with larger observations to recommend use of black Badri (indigenous) cows to improve blood picture. However, in India people are seen buying and taking goat milk vaguely once they are affected with Dengue fever.

Authors are thankful to the Vice Chancellor, Director Experiment Station, Dean, CVASc., Joint Director (IDF) and Head (LPM), CVASc. of G.B. Pant University of Agriculture and Technology, Pantnagar and AICRP-Goat Unit of ICAR-CIRG, Mathura at Pantnagar for providing necessary facilities to conduct the experiment, as a part of MVSc. research of the first author.