Hematological Assays Research Articles

P106: EARLY PREVENTIVE DIAGNOSIS OF HEMOLYTIC ANEMIA IN SICKLE CELL PATIENTS BY DETECTING THE TRIGGERING OF ERYPTOSIS.

Background/Aims: Sickle cell disease (SCD) also known as sickle cell anemia is one of the most worldwide-disseminated hereditary hemoglobinopathies. It is caused by a single amino acid substitution at the sixth residue of β globin gene (Glu6Val), which results in an abnormal hemoglobin called hemoglobin S (HbS). The acceleration of HbS polymerization induces rigid and dysfunctional erythrocytes that play a central role in acute and chronic clinical manifestations of SCD(Conran et al.,2009). Vaso-occlusion and hemolytic anemia are the hallmarks of SCD. We aim to explore the cellular environment of red blood cells to explain the physiopathology of SCD. In fact, the life span of circulating erythrocytes in healthy individuals vary from 100 to 120 days. In SCD, red blood cells undergo a form of cell death, namely, eryptosis before they reach their full life span. Eryptosis is triggered by a wide variety of factors as hyperosmolarity, oxidative stress and energy depletion. It is characterized by the presence of membrane blebbing, cell shrinkage, and phosphatidylserine (PS) exposure (Lang et al.,2012). In this study, we will explore the mechanism of triggering of eryptosis in Sickle cell disease. Methods: Following clinical diagnosis, 50 homozygous SCD patients and 30 healthy donors were identified for hematological and cellular assays. Flow cytometry was performed in order to determine the viability parameters of erythrocytes. The morphology of red blood cells and the externalization of phosphatidylserine was detected by labeling red blood cells with Annexine V. Moreover, we had identified intracellular calcium concentration and ceramide level by labeling erythrocytes with Fluo3-am and anti-ceramide antibodies. Finally, we had quantified reactive oxygen species (ROS) by labeling red blood cells with CM-H2DCFDA. Results and Discussion: Eryptosis in sickle cell patients is accelerated. In fact, PS (+) red blood cells are more present in patients than in healthy subjects. Therefore, eryptosis is triggered by oxidative stress, which stimulates the increase of calcium activity and subsequent externalization of PS and red blood cells shrinkage in sickle cell patients. However, the pathway of ceramide can also be considered a potential stimulating factor of eryptosis in SCD. Eryptosis ensures healthy erythrocyte quantity in circulation, whereas excessive eryptosis is the cause of acute anemia and may contribute to vaso-occlusive crisis in SCD patients.

Evaluation of Hepatic Enzyme Levels and Haematological Profiles in Chikungunya Patients- An Institutional Study

Background: Chikungunya is a viral infection caused by the Chikungunya virus (CHIKV) belonging to the Togaviridae family. The transmission of the virus is taking place through the bite of infected daytime biting female mosquitoes – primarily Aedes aegypti and Aedes albopictus. There were many reports which highlighted the number of reported cases of Chikungunya has been increasing in India in recent years, especially since 2016-2017. 
 The aim and objective of the present study was to understand the relationship between the severity of Chikungunya infection and its impact on levels hepatic enzymes and haematological profiles of individuals.
 Materials and Methods: Blood samples were collected from patients diagnosed with having Chikungunya. Routine laboratory investigations were carried out for analysing haematological assays like platelets, hematocrit count, haemoglobin count, total leukocyte count. and serum enzyme profiles includes Liver Function Test. 
 Result: Our results highlighted that the alterations of haematological and hepatic dysfunction are prevalent in Chikungunya infection, where haematological indices were significantly different between the control and patient group. RBC count, Hb, and WBC count were significantly lower in the patients than in the healthy control group. Liver enzymes like SGPT rising significantly more than SGOT. Hepatic Enzymes levels appear to have a directly proportional correlation with severity of infections. 
 Conclusion: Our study demonstrated that haematological and hepatic enzymes levels should be explored as routine laboratory markers for assessing the severity of Chikungunya infection, as they will help in employing an appropriate patient therapy, and thus optimise the use of available resources.

Effect of Fermentation with Rumen Content on the Feeding Value of Boiled Iron Tree (Prosopis africana) Seedcoat on Haematology and Serum Biochemistry of Broiler Chickens

Aims: The study was conducted to evaluate the effect of inclusion of Prosopis seed coat fermented with rumen content at different levels as energy source on the serum biochemical composition and heamatological indices of broiler chickens.
 Study Design:Data collected were subjected to analysis of variance in a completely randomized design.
 Place and Duration of Study: The study was conducted at the Livestock Teaching and Research Farm of the University of Agriculture Makurdi, Benue State, Nigeria. Between November 2016 and January 2017.
 Methodology: The study was in two phases; the starter and finisher phase, in each of the phases, there were five dietary groups and fermented Prosopis africana seed coat meal (FPASCM) was included at 0, 10, 15, 20 and 25% to replace maize. The total of two hundred and sixty (260), day- old Zartech broiler chicks were randomly distributed to five dietary treatments with four replicates of 52 chickens per treatment and each replicate had 13 birds. The birds were raised on deep litter and fed ad-libitum and drinking water provided daily and performance was monitored. At the end of the eight week, blood was collected via the wing web for serum and haematological assay.
 Results: The results of haematological parameters of broiler chickens fed varying levels of fermented Prosopis africana seed coat are presented in Table 3.There were no significant (P>0.05) differences between treatments for parameters measured except for PCV and haemogobin. The PCV values ranged from 25.00 -28.25% and haemoglobin values ranged from 8.35 -9.40g/dl. It was observed that most of the haematological indices measured fell within the normal reference range values. The results of serum biochemical assay revealed that serum glucose 45.85-107.22 mg/dl varied significantly (P<0.05) among the dietary treatments. Birds served 25% FPSCM had the highest value while those served 15% FPASCM recorded least value.
 Conclusion: Based on the observations from this study, it was concluded that FPASCM could be included in the diets of broiler chickens up to 25% without compromising the health of the birds.

The Acute Toxicity of Rutin in Mice

Acute toxicity is a step to evaluate the toxicity of a substance. Rutin is one of the flavonoid compounds with a variety of pharmacological effects. The aim of the study is to calculate the lethal dose that affect fifty percent of the mice used in the experiment (LD50). Thirty Swiss albino male and 30 non-pregnant female mice have been divided equally and randomly into 5 treated groups and one control group (n=5) Rutin has been administered with concentrations 5, 2.5.1.25,0.625 and 0.312 g/kg administered as a single dose intraperitoneally (IP) while the control group received 1% DMSO (IP). Animals were observed for any morbidity and mortality for 14 days. After 14 days the animal blood collected for biochemical and hematological analysis then all animals are euthanized for histopathological evaluation. The results showed the LD50 was 1.51 g/kg for male mice while for female mice was1.49 g/kg. No significant changes were observed at dose of 1.25glkg (female) and 0.625, 0.312 glkg (both sexes) in body weight measurements and in biochemical or hematological assays. Moreover no significant histopathological changes were reported compared to control.. It can be concluded that Rutin is practically a non-toxic substance.

Salsola imbricata Forssk. ameliorates acetic acid-induced inflammatory bowel disease by modulating dysregulated antioxidant enzyme system and cytokine signaling pathways in mice

Objective: To explore the protective effect of the crude extract of Salsola imbricata against acetic acid-induced inflammatory bowel disease in mice and its mechanism of action. Methods: Ethanolic crude extract of Salsola imbricata was characterized by HPLC. Salsola imbricata extract at different doses was administered and ulcerative colitis was induced by 200 μL, 7.5% acetic acid and macroscopic parameters were evaluated to assess the homeostatic condition of intestinal mucosa along with hematological and biochemical assays. The levels of malondialdehyde, glutathione peroxidase 1, superoxide dismutase, and catalase were determined in colon tissues. Proinflammatory cytokines including interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) were quantified by ELISA. The extent of tissue damage was assessed by histological analysis. Results: Phytochemical analysis confirmed the presence of phytochemicals including quercetin, gallic acid, syringic acid, benzoic acid and chlorogenic acid in the crude extract. The crude extract of Salsola imbricata (300 and 500 mg/kg) markedly decreased malondialdehyde and nitric oxide (P<0.01) and increased antioxidant activities of glutathione peroxidase 1 (P<0.001) and superoxide dismutase (P<0.001). Moreover, it decreased the levels of IL-1β, IL-6 and TNF-α significantly (P<0.001) and reduced the damage to the colon mucosa, promoting tissue healing and regeneration. Conclusions: Salsola imbricata extract restores the colonic epithelial layers by maintaining mucosal homeostasis and cell integrity by modulating antioxidant defense system and inflammatory cytokine signaling in ulcerative colitis mice.

Blood Performance: A New Formula for Fish Growth and Health

Simple SummaryThe use of haematological and blood biochemistry parameters has proven to be effective and repeatable ways to monitor fish health. Testing these parameters is becoming more common in aquaculture studies. Further, it is widely accepted that fish with better health status are more likely to grow faster as less energy should be consumed for non-growth purposes. Here, a new formula (Blood Performance) is introduced, which contains five common haematological and blood biochemistry parameters: red blood cells, white blood cells, haemoglobin, haematocrit, and total protein. The idea behind this formula is that any single component of this formula cannot be reliable enough as a biomarker of fish health and growth. However, interestingly, Blood Performance can be much more reliable and accurate for monitoring fish health and growth.Monitoring fish health in a repeatable and accurate manner can contribute to the profitability and sustainability of aquaculture. Haematological and blood biochemistry parameters have been powerful tools and becoming increasingly common in aquaculture studies. Fish growth is closely related to its health status. A fish with a higher growth rate is more likely to be a healthy one. Any change in the physiological status of the fish, from pollution to nutritional stress, can cause changes in the blood parameters. Various aquaculture studies have measured the following components: red blood cells, white blood cells, haemoglobin, haematocrit, and total protein. However, because these parameters do not always follow the same trend across experimental fish, it is difficult to draw a firm conclusion about which parameter should be considered. Therefore, Blood Performance (BP) as a new formula is introduced, which is a more reliable indicator. This formula is simple and sums up the natural logarithm of the five above-mentioned parameters. More than 90 published peer-reviewed articles that measured these five parameters in the last six years confirmed the reliability and validity of this formula. Regardless of which supplements were added to the diets, the fish with a higher growth rate had higher BP as well. In addition, in 44 studies out of 53 articles, there was a significant positive correlation between specific growth rate and BP. Under different stressful situations, from pollution to thermal stress, the fish under stress had a lower BP than the control. Fish meal and fish oil replacement studies were further evidence for this formula and showed that adding excessive alternative proteins decreased growth along with BP. In conclusion, BP can be a reliable indicator of fish health and growth when it is compared between groups in the same experiment or farm. Although there was a positive correlation between specific growth rate and BP, comparing BP between experiments is not recommended. Standardising the haematological assays can improve the reliability and accuracy of BP across experiments.

Assessing the Physiologic Relevance of Red Blood Cell Deformability in Iron Deficiency Anemia

Background: Iron deficiency anemia (IDA), which affects individuals of all ages worldwide, is an often overlooked and undertreated component of chronic disease, despite data correlating its association with adverse outcomes in patients with cardiovascular disease (von Haehling, Nat Rev Cardiol, 2015). While red blood cells (RBCs) in IDA are known to be smaller and contain less hemoglobin than healthy RBCs, how RBC deformability is altered in IDA remains poorly understood; some ektacytometry studies have observed impaired deformability in iron deficient RBCs (idRBCs), while others described either unchanged or increased deformability (Brandão, Clin Hemorheol Microcirc, 2009). Here we ask: can single cell biophysical techniques definitively determine whether idRBCs are less deformable than healthy RBCs and how heterogenous that phenomena may be? Recent investigations into IDA's role in cardiovascular disease have generally focused on the myocardium and coronary vasculature, yet much regarding other physiologic implications remains unknown, including whether idRBCs cause microvascular obstruction or vasculopathy. To address such questions, we leveraged a suite of microvascular models we developed.Methods: We first coupled our microfluidic capillary model with μEXACT, our customized automated particle tracking program for hematologic cell-based assays, to collect high-throughput velocity tracking of single RBCs from a healthy control and 2 IDA patients (anemic for age, ferritin <10 ng/mL) to create a single cell deformability index (sDI) for each RBC (Fig 1). Next, whole blood samples collected in EDTA tubes from the control and IDA patients were perfused into both straight 100μm wide channels (mimicking large venules) and branching 30μm wide microfluidic devices (mimicking smaller venules) at a constant shear rate for 30 minutes to observe if any occlusions or obvious alterations in flow were observed (Fig 2). Finally, using the straight 100μm channel microfluidic devices, human umbilical vein endothelial cells (HUVECs) were cultured throughout each microchannel and RBCs from a healthy control and 3 IDA patients were perfused in parallel microchannels for 4 hours. The endothelialized models were then fixed, permeabilized, and immunostained with antibodies against VCAM-1 and E-selectin, known markers of endothelial inflammation. Mean fluorescence intensity was measured to quantify endothelial inflammation (Fig 3).Results: sDI distribution histograms were obtained for healthy and IDA patient RBCs. The mean sDIs for IDA patient RBCs were decreased in comparison to the healthy RBCs. Additionally, both IDA patient's RBCs lacked a subpopulation of highly deformable RBCs, likely reticulocytes, seen in the healthy RBCs (Fig 1C). There was no evidence of microchannel occlusion for the healthy control or IDA patient whole blood samples in either the straight 100μm microchannels or branching 30μm microfluidic devices (Fig 2D). Finally, in our endothelialized microfluidic model, endothelium exposed to IDA patient RBCs exhibited increased VCAM-1 and E-selectin expression over endothelium exposed to healthy RBCs (Fig 3B).Conclusions: By utilizing an array of microfluidic models we can develop a more comprehensive understanding of the role idRBCs play systemically on microvasculature. Our combined microfluidic and image analysis system demonstrated decreased deformability in idRBCs and can offer detection of subpopulation differences that cannot be fully characterized with bulk techniques such as ektacytometry. So far, our data demonstrates that while no microvascular occlusion occurs, idRBCs contribute to endothelial inflammation. Additionally, the observation that physical interactions between endothelial cells and idRBCs are sufficient to cause endothelial inflammation warrants further investigation, as generally idRBCs had not been viewed as pro-inflammatory. Ongoing studies will couple unique sDI distribution curves with the degree of endothelial inflammation, as well as elucidate how these changes are associated with the degree of IDA or clinical events such as the initiation of iron supplementation. Utilizing atomic force microscopy to better understand how the idRBC membrane impacts deformability and developing biophysical computer simulations to determine if increased idRBC-endothelium interactions are observed in silico are also planned. [Display omitted] DisclosuresLam: Sanguina, Inc.: Current holder of individual stocks in a privately-held company.

The effects of quercetin on seizure, inflammation parameters and oxidative stress in acute on chronic tramadol intoxication

BackgroundTramadol is a widely used synthetic opioid for moderate to severe pain. Some studies have shown that tramadol can increase oxidative stress in different tissues of the body. Quercetin is also a substance with various biological effects, including antioxidant, anti-inflammatory, hepatoprotective, nephroprotective, and cardioprotective activities. The current investigation aimed at determining the effects of quercetin, with or without naloxone, on tramadol intoxication.MethodsThis study was performed on 30 male Wistar rats divided into five groups: Group I) control group: intraperitoneal injections of normal saline 0.9% for 14 days; Group II) tramadol: 25 mg/kg for 14 days, and then a 50 mg/kg acute dose injection on the last day; Group III) acute quercetin (single dose): tramadol injection as with the second group plus 100 mg/kg of quercetin on the last day; Group IV) chronic quercetin: tramadol injection similar to the second group plus quercetin 100 mg/kg for 14 days; Group V) quercetin plus naloxone: tramadol injection similar to the second group plus injection of quercetin 100 mg/kg + intravenous naloxone 2 mg/kg on the last day, followed by a 4 mg/kg/h injection of naloxone for six hours. The rats were monitored for six hours on the last day, relating to the number and severity of seizures. Finally, the samples were prepared for biochemical investigation of the serum level of oxidative stress markers (MDA, SOD, NOx), inflammatory factors (IL-6, TNF-α), biochemical parameters (ALT, AST, creatinine, glucose) and hematological assay. The liver, heart, kidney, cortex, cerebellum, and adrenal tissues were collected to investigate the redox state.ResultsNone of the treatments had positive effects on the number and severity of seizures. Chronic administration of quercetin led to alteration of some blood parameters, including reduced hemoglobin level and elevated platelet counts. Acute on chronic tramadol administration resulted in a significant rise in AST, where different treatments failed to reduce their levels down to the control group.Conclusionchronic administration of quercetin showed decreased oxidative/nitrosative stress in the liver, kidney, adrenal, and heart tissues. Quercetin plus naloxone decreased oxidative stress in the heart and adrenal tissues, but adverse effects on the brain cortex and hepatic function. Single-dose quercetin reduced cardiac oxidative stress.

Indices in differentiating iron deficiency anemia from thalassemia trait- a comparative study

Background: Iron deficiency anemia (IDA) and beta thalassemia trait (BTT) are the two most common and important causes of microcytic hypochromic anemia in India. It is very difficult to differentiate between the two. Many different types of techniques have been proposed for the same. While some are invasive like bone marrow examination others are not available at all centers, like electrophoresis. Hence different indices come into play. Aims and Objective: This study was undertaken to compare the efficacy of Shine and Lal index and Mentzer index in differentiating between IDA and BTT. Materials and Methods: A total of 407 anemia cases were studied over a period of 18 months and their blood samples were subject to different hematological and biochemical assays to diagnose the type of anemia. Results: Based on these tests 92.1% cases were found to be of IDA whereas 3.7% cases were found to be of BTT. Then both the indices were applied in the above mentioned cases. Conclusion: While Shine and Lal index was found to have better sensitivity, Mentzer index was found to have better specificity.

Ameliorative effect of ink extract and polysaccharide of Sepia officinalis on hepatotoxicity, renal toxicity and hematological disorders in adult male albino rats treated with cyclophosphamide

This study aimed to investigate ameliorative effects of the crude extract (SIE) and polysaccharide (SIP) of the ink of Sepia officinalis, on some biochemical and hematological disorders induced by cyclophosphamide (CP). Forty adult male albino Wistar rats were divided into five groups (n= 8 each). In the control group, rats were administered orally with 0.9% isotonic saline solution at a dose (5 ml/kg b.w.). All the other groups were i.p. injected with a single dose of CP (200mg/kg b.w.) only for one time. Then the third group was treated with oral administration of (SIE) (200mg/kg b.w.) daily for 60 days, the fourth group was treated with oral administration of (SIP) (80mg/kg b.w.) daily for 60 days and the fifth group was treated with oral administration of (SIE, 200mg/kg b.w. + SIP, 80mg/kg b.w.) daily for 60 days. All the animals were slaughtered by the end of the experiment for collecting the blood samples for hematological and biochemical assays. The biochemical results indicated that administration of CP was associated with a significant increase in the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea, creatinine and uric acid. Moreover, a significant decrease in the levels of albumin and total protein was recorded. In addition, hematological disorders including a significant suppression on the numbers of RBCs, WBCs and PLTs, with a remarkable reduction in hemoglobin contents (Hb) and a significant drop in PCV values. Concomitant administration of SIE and SIP alleviated the altered biochemical and hematological parameters.

Hematological and kidney-functional analysis of acetic acid-induced inflammatory rats administered flavonoid-rich fraction of Ficus sur

Background and aims: Unregulated inflammation causes dysfunctional immune system and could consequently lead to blood disorders due to impaired hematopoietic process, and also destruction of important organs as a result of autoimmunity. In this study, flavonoid-rich fraction of Ficus sur (FRFFS) is investigated for its effects on hematological parameters and renal metabolites in inflammation-induced rats. Materials and methods: Inflammation was induced in Wistar rats of average weight of 112.60 ± 0.75 g by successive oral administration of 6% at (20 ml/kg), 3% at (15 ml/kg) and 6% at (10 ml/kg) acetic acid for one week. Experimental animals were orally administered various doses of FRFFS; 15 mg/kg, 30 mg/kg, 60 mg/kg and 120 mg/kg for two weeks. Blood and kidney samples were collected for hematological parameters and kidney functional assays respectively, and data were analyzed using SPSS version 20. Results: FRFFS significantly (p &lt; 0.05) increased pack cell volume, hemoglobin, red blood cells and platelets, and significantly decreased white blood cells, most especially at 120 mg/kg and 60 mg/kg doses. Similarly, it significantly increased kidney urea and creatinine concentrations. Conclusion: Sequel to these findings, it is inferred that FRFFS could be a potential natural therapy for hematological and renal disorders.

Hematological and Histopathological Profiles of Swiss Albino Mice Infected with Plasmodium berghei upon Treatment with Cooked Tetracarpidium conophorum Seed Ethanol Extract

Aim: This study was conducted to verify the hematological and histopathological outcome of cooked ethanol seed extract of Tetracarpidium conophorum in swiss albino mice infected with Plasmodium berghei (NK65).&#x0D; Study Design: Descriptive study.&#x0D; Methodology: Standard methods were conducted to determine the phyto-chemical profile of cooked T. conophorum seed ethanol extract, hematological, histopathological indices and biochemical assay. Mice were assembled into five groups (1, 2, 3, 4 and 5) of seven each. The mice in group 4 were treated with a customary antimalarial drug (chloroquine as positive control) at a dose of 5 mg/kg body weight, while mice in groups 3, 2 and 1 was administered with increasing dosages (200, 400, 800 mg/kg body weight) of seed extracts for four consecutive days respectively.&#x0D; Results: The qualitative phytochemical screening of T. conophorum seed extract revealed the presence of bioactive components such as saponin, tannins, flavonoid, terpenoid. Saponin (16.82 mg/100 g) was the highest occurring phytochemical followed by terpenoid (10.39 mg/100 g), glycoside (5.9 mg/100 g), tannin (2.8 mg/100 g) and flavonoid (1.17 mg/100 g), while steroid only had a trace presence. Hematological parameters revealed mice in all groups do not possess significantly different mean corpuscular haemoglobin concentration (MCHC), mean corpuscular hemoglobin (MCH) and mean corpuscular volume (MCV). Mice in group 5 (infected and not treated) demonstrated lowest values of hemoglobin, platelet, and packed cell volume with a significantly lower % eosin. There was considerable decrease in the levels of alanine transaminase and aspartate transaminase in group 5 compared to mice in groups 1, 2, and 3.&#x0D; Conclusion: Ameliorative effects of seed extract were observed on the liver and kidney of mice at dose levels (200 mg/kg) used, but the 400 mg/kg restored the kidney but had adverse effect on the liver. This study therefore shows that cooked T. conophorum is generally less effective and could not be employed for treatment of malaria.

Prevention of doxorubicin-induced haematotoxicity by turmeric in Wistar rats

Doxorubicin is a known anticancer drug used in treatment of various cancers including leukemias, lymphomas, soft-tissue sarcomas and solid tumors. Doxorubicin exerts its toxicity on the blood by causing bone marrow suppression, myelosuppression, pronounced marrow depression and bone marrow depletion, and hypoplasia This study investigated the effect of turmeric on the haematological parameters in doxorubicin-induced oxidative stress in Wistar rats. 54 adult Wistar rats were divided into 9 groups of six animals each. Group 1 animals served as control (normal saline), group 2 animals served as negative control, and received doxorubicin, group 3 animals were given doxorubicin and turmeric, group 4 animals received doxorubicin and vitamin C, group 5 animals received doxorubicin and vitamin E, group 6 animals received doxorubicin, vitamins C and turmeric, group 7 animals received doxorubicin, vitamin E and turmeric, while group 8 animals received doxorubicin, vitamin C and vitamin E and finally, group 9 animals receive doxorubicin, vitamin C, vitamin E and turmeric. The experiment lasted for 28 days and blood samples were collected from each animal from the various groups for haematological assay. Doxorubicin caused a significant decrease in the serum levels of white blood cell count, neutrophil count, lymphocyte count, monocyte count, platelet count, red blood cell count, hemoglobin concentration, hematocrit and mean corpuscular volume. These changes were prevented by turmeric alone or in combined concomitant administration with vitamins C and or E and doxorubicin

Haematology of captive wild African giant (Cricetomys gambianus) and brown (Rattus norvegicus) rats sero-positive for Leptospira spp. around Zaria

In this study, the haematology of captive wild African giant (Cricetomys gambianus) and brown (Rattus norvegicus) rats sero-positive for Leptospira spp. around Zaria was determined. Blood was collected by intracardiac aspiration using chloroform anaesthesia from 10 adult wild rats (5 African giant and 5 brown rats) caught within Zaria and its environs. The individual blood samples were screened for antibodies against Leptospira spp. using IgG ELISA kit. Blood samples of rats positive to Leptospira spp. were subjected to haematological assay carried out following standard procedures. The sero-prevalence of antibodies against Leptospira spp. in both rat species was found to be 100%. In conclusion, the monocyte count in the African giant rat were significantly lower (P = 0.000) compared to corresponding values in the brown rat.

Growth performance and haematological traits of Nigerian local chickens fed varied dietary protein levels

This study was carried out to investigate the growth performance and haematological traits of Nigerian local chickens (NLC) fed varied dietary protein levels. An eleven week feeding trial was done in a 3 x 2 factorial arrangement using six treatment groups comprising a combination of two genotypes of the NLC' identified as normal leather (na) and naked neck (Na) and three dietary protein levels - high protein: HP (22% and 20% CP chick mash and grower mash respectively), medium protein: MP (20% and 18% chick mash and grower mash respectively) and low protein: LP (18% and 16% chick mash and grower mash). Data were collected on growth performance traits (body weight, feed intake, feed conversion ratio and protein efficiency ratio). At the end of the feeding trial a haematological assay was done. Neither genotype nor interaction of genotype and dietary protein level influenced (p&gt;0.05) any of the growth performance traits studied. Birds on HP ate 10.21% and 7.09% less feed than those on MP and LP respectively. A better (p&lt;0.05) protein efficiency ratio (up 10 20.79%) was observed in birds fed LP. Na birds had 4.55% and 17.50x704L-1 more packed cell volume and platelets respectively than na birds (p&lt;0.05). All haematological traits studied, except white blood cell count, showed significant variations (p&lt;0.05) due to varied dietary protein levels. Higher haemoglobin, red blood cell counts, mean corpuscular haemoglobin concentration and platelets were observed in birds fed LP diet. Reducing the dietary protein level had no adverse effect on the NLC thus, the LP diet is recommended for raising the Nigerian local chicken.

Utilization of coconut meal based diets supplemented with lysine by broiler chicken

Coconut meal (CM) was included in a broiler diet to supply 40% of the total groundnut cake protein. The diet was formulated based on the recommendations of a previous study that coconut meal protein can replace 40% of groundnut cake (GNC) in broilers diet. A control diet was formulated containing 22% GNC, the protein of which was replaced by coconut meal protein. Five diets were prepared . The control (Diet A) and 4 others that contained essentially the same proximate composition (Diets B-E) were supplemented with synthetic lysine at 0.30, 0,60, 0.90 and 1.20% dietary levels. 240 broiler chicks were alotted into 5 dietary treatment groups of 3 sub-groupa. Each sub-group contained 16 birds. The dietary lysine content from diets A-E were 1.20, 1.15, 1.45, 1.76 and 2.00% respectively and were fed to the broilers for a period of 56 days. Performance indices, average feed consumption, average body gain, live weight, feed conversion ratio (FCR), protein efficiency ratio (PER) were monitored. The dry matter, protein and ether extractretentions were all equally monitored twice at the 4th and 8th week. Haematological and enzyme assays were investigated at 4th and 8th week. The average feed consumption and FCR were significantly increased (P&lt;0.05) by the increasing level of lysine supplementation both at the 4th and 8th week. The body gain also decreased significantly (P&lt;0.05) at the 4th and 8th week while the live weight was significantly (P&lt;0.05) better in the control and 0.6% lysine supplementation at the 56th day of the study. The dry matter, nitrogen and other extract retentions significantly decreased (P&lt;0.05) at the 4th and 8th week. The total protein and albumin decreased significantly (P&lt;0.05) at the 4th and 8th week while, creatinine and urea increased with increasing level of lysine supplementation. The serum glutamate amino transaminase and alanine amino transaminase both increased significantly (P&lt;0.05) at the 4th and 8th week. The study indicated that 0.6% lysine supplementation (of diet containing coconut meal contributing 40% protein of the major vegetable protein source) enhance the utilization of the coconut meal by broilers.

Growth performance and haematological traits of Nigerian local chickens fed varied dietary protein levels

This study was carried out to investigate the growth performance and haematological traits of Nigerian local chickens (NLC) fed varied dietary protein levels. An eleven week feeding trial was done in a 3 x 2 factorial arrangement using six treatment groups comprising a combination of two genotypes of the NLC identified as normal feather (na) and naked neck (Na) and three dietary protein levels - high protein: HP (22% and 20% CP chick mash and grower mash respectively), medium protein: MP (20% and 18% chick mash and grower mash respectively) and low protein: LP (18% and 16% chick mash and grower mash). Data were collected on growth performance traits (body weight, feed intake, feed conversion ratio and protein efficiency ratio). At the end of the feeding trial a haematological assay was done. Neither genotype nor interaction of genotype and dietary protein level influenced (p&gt;0.05) any of the growth performance traits studied. Birds on HP ate 10.21% and 7.09% less feed than those on MP and LP respectively. A better (p&lt;0.05) protein efficiency ratio (up to 20.79%) was observed in birds fed LP. Na birds had 4.55% and 17.50x103 L-1 more packed cell volume and platelets respectively than na birds (p&lt;0.05). All haematological traits studied, except white blood cell count, showed significant variations (p&lt;0.05) due to varied dietary protein levels. Higher haemoglobin, red blood cell counts, mean corpuscular haemoglobin concentration and platelets were observed in birds fed LP diet. Reducing the dietary protein level had no adverse effect on the NLC thus, the LP diet is recommended for raising the Nigerian local chicken.

Wild epiphytic Bangladeshi orchids Cymbidium aloifolium (L.) Sw. and Papilionanthe teres (Roxb.) Lindl. potentially modulates the immune functions in Swiss albino mice.

Objective:This research investigated the immunomodulatory potentials of two medicinally important wild epiphytic Bangladeshi orchids Cymbidium aloifolium and Papilionanthe teres using Swiss albino mice.Materials and Methods:Orchid extracts were prepared using a cold methanol extraction procedure. To assess the immunomodulatory action, Swiss albino mice of either sex weighing 25–35 gm were divided into five groups each with six animals. Sheep red blood cells (SRBC) of 0.5 × 109 cells/ml were used to immunize all mice on the 7th day, and a booster dose of the same quantity of SRBC was given on the 11th day of the experiment. After 14 days of oral treatment with 100 and 200 mg/kg bw of orchid extract, the mice were sacrificed to collect serum and organs. Hematological assays, delayed type of hypersensitivity assays, phagocytic index (PI), and histopathological investigations were used to assess in vivo immunomodulatory efficacy.Results:The body weight changes of the experimental animals were considerably greater at 100 mg/kg bw than at a higher dose (200 mg/kg bw). There was a substantial improvement of relative organ weights of the thymus and spleen at the low dose, but no effect on kidney weights was evident. The liver weight increased significantly (p < 0.05) at both doses. Total neutrophil, leukocyte, and lymphocyte counts, hemoglobin percentage, delayed hypersensitivity reaction, and PI were all significantly (p < 0.05) increased in mice receiving the lower dose. In contrast to the control group, the higher dose reduced immunological response, suggesting the negative influence of a higher dose of extracts on the immune reaction.Conclusions:The results demonstrate that orchid extracts can potentially modulate the innate immune system in the experimental animal.

Oxidative stress in Strongylus spp. infected donkeys treated with piperazine citrate versus doramectin.

Background: Parasitic infection is one of the main problems in equidae, particularly donkeys. Aim:This study evaluated the oxidative stress in donkeys infected with Strongylus spp by determining the correlation between antioxidants levels; malondialdehyde (MDA), total antioxidant capacity (TAC), and the severity of parasitic infection. It also compared the therapeutic efficacy of piperazine citrate as an oral anthelmintic drug and Doramectin as an injectable one. Methods:The study was conducted on 40 donkeys naturally infected with Strongylus spp. These donkeys were divided into two groups (20 donkeys each) according to treatment; One group was treated with piperazine citrate (PipTG) and the other with doramectin (DoraTG). Thorough clinical examination, hematological, biochemical, and parasitological assays were performed before (Day 0) and after treatment (Days 7, 14, 21, and 28). All data were statistically analyzed by independent-sample t-test or paired t-test. Results:In both groups, mean values of MDA were significantly reduced, while those of TAC were significantly elevated after treatment on days 7, 14, 21, and 28. These significant changes were reported after treatment between PipTG and DoraTG in favor of DoraTG. Serum concentrations of MDA were significantly reduced, while those of TAC were significantly elevated for DoraTG treatment group when their values were compared with those of PipTG either on days 7, 14, 21, or 28. Significant correlations were reported in PipTG and DoraTG. Negative significant correlations were reported between fecal egg count (FEC) and each of whole blood picture indices (RBCS, Hb, and PCV), serum TAC and faecal egg count reduction percentage FECR%. A positive correlation was seen between FEC and MDA. MDA exhibited a negative correlation with both blood picture and TAC; hence, TAC was positively correlated with these blood picture indices in both PipTG and DoraTG. In PipTG, anthelmintic resistance (R) was present on days 7 and 14, while it was suspected (S) at day 21 then it was absent (N) at day 28. In DoraTG, anthelmintic resistance was suspected (S) on day 7, then it became absent (N) on days 14, 21, and 28 post therapy. Conclusion:The immunological status of the infected donkeys had greatly improved after treatment. The therapeutic efficacy of injectable doramectin was more efficient than that of oral piperazine citrate in Strongylus spp. infected donkeys.

Safety of oral administration of high doses of ivermectin by means of biocompatible polyelectrolytes formulation

The FDA-approved drug ivermectin is applied for treatments of onchocerciasis and lymphatic filariasis. The anti-cancer and anti-viral activities have been demonstrated stressing possibilities for the drug repurposing and therefore new information on high dosage safety is on demand. We analyzed in vivo tissue responses for high doses of ivermectin using Corydoras fish as animal model. We made intestinal histology and hematologic assays after oral administration of ivermectin transported with polyelectrolytes formulation. Histology showed any apparent damage of intestinal tissues at 0.22–170 mg of ivermectin/kg body weight. Immunofluorescence evidenced delocalization of Myosin-Vb at enterocytes only for the higher dose. Hematology parameters showed random variations after 7 days from administration, but a later apparent recover after 14 and 21 days. The study evaluated the potential of high doses of oral administration of ivermectin formulation, which could be an alternative with benefits in high compliance therapies.