Direct blood PCR detection of Babesia bigemina and its effect on haematological and biochemical profile in crossbred cattle of eastern Haryana

2017 ◽  
Author(s):  
Anita Ganguly ◽  
R. S. Bisla ◽  
Indrajit Ganguly ◽  
Harpreet Singh ◽  
Vandna Bhanot ◽  
...  
Keyword(s):  
Clinical Signs ◽  
Pcr Detection ◽  
Biochemical Profile ◽  
Crossbred Cattle ◽  
Babesia Bigemina ◽  
Direct Pcr ◽  
Blood Samples ◽  
Crossbred Cows ◽  
Specificity And Sensitivity ◽  
Healthy Control

The present study aimed to diagnose Babesia bigemina in naturally infected crossbred cows and to determine its effect on haemato-biochemical profile of host animals. Blood samples from lactating crossbred cows (n=30) between 3-6 years of age and showing clinical signs of babesiosis were collected, with or without anticoagulant, and analyzed for the protozoa by direct smear, direct blood PCR detection of the apical membrane antigen 1 (AMA-1) gene specific amplicon of B. bigemina and estimation of haematological and biochemical parameters. Healthy crossbred cows (n=10), examined free from haemoprotozoan infections were included as control. Blood Direct PCR revealed a 448-bp amplified fragment. Out of 150 random blood samples screened, (27/150) 18% were positive under light microscope, whereas direct blood PCR revealed (39/150) 26% samples positive for B. bigemina. The result shows higher specificity and sensitivity of PCR test over blood smear examination. The infected group showed significantly (p less than 0.001) decreased levels of TEC (3.04±0.19), Hb (4.78±0.27) and PCV (14.53 ±0.87) than healthy control animals. However, differences in the red blood cell indices (MCV, MCH and MCHC) were non-significant (p>0.05) between the groups indicating normocytic hypochromic anaemia in affected crossbred cattle. Serum samples of infected cows showed significantly (p less than 0.01) higher values of ALT (78.83±8.95), AST (146.13±7.62), BUN (27.09±1.02), creatinine (1.93±0.1) and TBIL (1.42±0.06) than that of healthy control. A significant decrease (p less than 0.01) of TSP (6.12±0.13) and albumin (2.39±0.09) was also recorded in the infected cows compare to healthy control. The standardized blood direct PCR method of the present investigation may be useful for rapid and reliable diagnosis of B. bigemina in conjunction with microscopic examination. Moreover, marked changes in haematological and serum biochemical profile observed in B. bigemina infected crossbred cows may be useful in understanding disease pathogenesis and undertaking necessary corrective measures..

Investigations on First Confirmed Outbreak of Ovine Theileriosis (Theileria luwenshuni) from Maharashtra State, India

2020 ◽  
Author(s):  
V.S. Dhaygude ◽  
K. Kundu ◽  
B.P. Kamdi ◽  
U.R. Bagal ◽  
S.B. Bhosale ◽  
...  
Keyword(s):  
18S Rrna ◽  
Clinical Signs ◽  
Specific Treatment ◽  
Small Ruminants ◽  
18S Rrna Gene ◽  
Pcr Detection ◽  
Nucleotide Sequencing ◽  
Rrna Gene ◽  
Blood Samples ◽  
Theileria Luwenshuni

Background: Clinical theileriosis of small ruminants is tick-borne disease caused by Theileria lestoquardi, Theileria uilenbergi and Theileria luwenshuni. Theileria annulata, the causative agent of bovine tropical theileriosis in cattle, can also infect sheep but does not cause any significant illness. It is one of the economically important diseases. There are no reports of ovine clinical theileriosis from Maharashtra state and there is paucity of information on its epidemiology. This paper reports first confirmed outbreak of ovine theileriosis based on clinical signs, microscopic examination, PCR and sequencing in the Maharashtra State of India. Methods: Whole blood samples from 22 ailing sheep were collected and subjected to hematological examination. Blood smears stained with Leishman’s stain were examined under 100X objective of the microscope. The blood samples from sheep found positive by microscopic method were subjected to PCR detection of 18S rRNA gene of hemoprotozoa and then for nucleotide sequencing and sequence analysis.Conclusion: Samples from 14 out of 22 sheep were found positive for piroplasms of Theileria spp by light microscopy. All positive samples were further confirmed by PCR detection of 18S rRNA gene of hemoprotozoa. PCR amplification yielded expected product of 1750 bp for all samples. BLAST and phylogenetic analysis of one sample revealed high sequence homology with T. luwenshuni reported from India and other countries. Characteristic clinical signs like fever, progressive anaemia, laboured breathing, lymphadenopathy, debility and non-responsiveness to antibiotic therapy were recorded. The animals responded to specific treatment against theileriosis. It is the first ever confirmed report of ovine theileriosis in Maharashtra state of India and hence reported.

ALTERATIONS IN SOME BIOCHEMICAL PARAMETERS IN CATTLE AFFECTED WITH FOOT AND MOUTH DISEASE IN DAKAHLIA GOVERNORATE, EGYPT

2018 ◽  
pp. 49-56
Author(s):  
Eman Kamal ◽  
MOhamed Salama ◽  
Ahmed Elgamal ◽  
Nabil Heakal
Keyword(s):  
Biochemical Parameters ◽  
Serum Insulin ◽  
Clinical Signs ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Control Group ◽  
Infected Cattle ◽  
Blood Samples ◽  
Foot And Mouth ◽  
Healthy Control

This study was carried out to evaluate the changes in serum biochemical parameters during foot and mouth disease (FMD) viral infection, so blood samples were collected from different farms in Dakahlia Governorate during the outbreak of the disease in 2016. Blood samples were collected after complete clinical and physical examination of twenty five cattle; their ages were ranging from 1-2 years and divided into two groups as follows: 15 cattle were showing the clinical signs of the disease and 10 cattle were clinically healthy (control group). The obtained results showed a significant increase in glucose level (P<0.01), non-esterified fatty acids (NEFA) (P<0.001), beta hydroxyl butyric acid (BHBA) (P<0.001) and lipase enzyme (P<0.01) in infected cattle compared to control group. On the other hand, serum insulin concentration and amylase were significantly lower in blood of infected cattle than control group (P<0.001) and (p<0.01) respectively. from the obtained results it could be concluded that FMD infection in cattle is associated with hyperglycemia, hypoinsulinemia and ketosis

Association of Retained Fetal Membranes with Haematological Profile in Crossbred Cows

2017 ◽  
Vol 12 (3) ◽  
Author(s):  
K. Rokde ◽  
S. Kumar ◽  
A. Bhardwaz ◽  
S. S. Mahour ◽  
S. P. Nema ◽  
...  
Keyword(s):  
Eosinophil Count ◽  
Jugular Vein ◽  
Dairy Farm ◽  
Fetal Membranes ◽  
Monocyte Count ◽  
Blood Samples ◽  
Mean Values ◽  
Crossbred Cows ◽  
Haematological Profile ◽  
The Mean

This study was carried out on clinical cases of retained fetal membranes in crossbred cows presented at College Clinics and College dairy farm and from Villages in and around Mhow. The blood samples were collected from jugular vein just before 12 hr. postpartum and on 7th day postpartum. Haematological profile revealed that the mean values of haemoglobin, neutrophil and monocyte count after 12 hrs and 7th day postpartum were significantly lower and lymphocyte count was significantly higher in RFM cows (n=18) than normally calved cows (n=6). The differences in mean TLC, eosinophil and basophil counts were non-significant at 12 hrs postpartum, however on 7th day postpartum the TLC and eosinophil count were significantly higher and basophil count was non-significantly different in RFM cows than the normally calved cows.

scholarly journals Development of a Co-Dominant Cleaved Amplified Polymorphic Sequences Assay for the Rapid Detection and Differentiation of Two Pathogenic Clarireedia spp. Associated with Dollar Spot in Turfgrass

Agronomy ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1489
Author(s):  
Tammy Stackhouse ◽  
Sumyya Waliullah ◽  
Alfredo D. Martinez-Espinoza ◽  
Bochra Bahri ◽  
Emran Ali
Keyword(s):  
Pcr Primers ◽  
The United States ◽  
Fungal Species ◽  
Dollar Spot ◽  
Direct Pcr ◽  
Specific Pcr ◽  
Specificity And Sensitivity ◽  
Pure Cultures ◽  
Speed Up ◽  
Cleaved Amplified Polymorphic Sequences

Dollar spot is one of the most destructive diseases in turfgrass. The causal agents belong to the genus Clarireedia, which are known for causing necrotic, sunken spots in turfgrass that coalesce into large damaged areas. In low tolerance settings like turfgrass, it is of vital importance to rapidly detect and identify the pathogens. There are a few methods available to identify the genus Clarireedia, but none of those are rapid enough and characterize down to the species level. This study produced a co-dominant cleaved amplified polymorphic sequences (CAPS) test that differentiates between C. jacksonii and C. monteithiana, the two species that cause dollar spot disease within the United States. The calmodulin gene (CaM) was targeted to generate Clarireedia spp. specific PCR primers. The CAPS assay was optimized and tested for specificity and sensitivity using DNA extracted from pure cultures of two Clarireedia spp. and other closely related fungal species. The results showed that the newly developed primer set could amplify both species and was highly sensitive as it detected DNA concentrations as low as 0.005 ng/µL. The assay was further validated using direct PCR to speed up the diagnosis process. This drastically reduces the time needed to identify the dollar spot pathogens. The resulting assay could be used throughout turfgrass settings for a rapid and precise identification method in the US.

scholarly journals Evaluation of a Preclinical Blood Test for Scrapie in Sheep Using Immunocapillary Electrophoresis

2006 ◽  
Vol 89 (3) ◽  
pp. 720-727 ◽  
Author(s):  
Roy Jackman ◽  
David J Everest ◽  
Mary Jo Schmerr ◽  
Mohammed Khawaja ◽  
Pat Keep ◽  
...  
Keyword(s):  
Capillary Electrophoresis ◽  
Prion Protein ◽  
Clinical Signs ◽  
Test System ◽  
Buffy Coat ◽  
Infected Sheep ◽  
Test Method ◽  
Peptide Sequence ◽  
Transmissible Spongiform Encephalopathies ◽  
Blood Samples

Abstract An analytical method is described for detection of endogenous disease-associated prion protein in the buffy coat fraction from the blood of sheep infected with scrapie. The method has been improved and evaluated for its performance in the preclinical diagnosis of ovine transmissible spongiform encephalopathies. The test system uses a protocol for sample preparation that includes extraction and concentration and a test method that uses a liquid-phase competitive immunoassay for prion protein. Antibodies directed to a peptide sequence at the C-terminus of the prion protein (PrP) and a fluorescein-labeled peptide conjugate are used in the assay. Free zone capillary electrophoresis with laser-induced fluorescence for detection is used to separate the antibody-bound fluorescently labeled peptide and free labeled peptide. In this assay, the PrP competes with the fluorescently labeled peptide for limited antibody binding sites, which results in a reduction of the peak representing the immunocomplex of the antibody bound to the fluorescently labeled peptide. When blood samples from scrapie-infected sheep aged 712 months and of the scrapie-susceptible PrP genotypes VRQ/VRQ and VRQ/ARQ were analyzed, the abnormal PrP was found in blood samples. These results correlated with the post-mortem diagnosis of scrapie. The sheep were preclinical and appeared normal at the time of testing but later died with clinical disease approximately 12 months after testing. In older animals, and those with clinical signs, a smaller percentage of animals tested positive. This study has demonstrated that this technology can be used as a sensitive, rapid preclinical test to detect the disease-associated PrP in the blood of scrapie-infected sheep. Improvements in the extraction protocol and capillary electrophoresis conditions will enhance the robustness of this test.

scholarly journals Diagnosis and Treatment of Human Salmonellosis in Addis Ababa City, Ethiopia

2018 ◽  
Vol 2018 ◽  
pp. 1-7
Author(s):  
Legesse Garedew ◽  
Semaria Solomon ◽  
Yoseph Worku ◽  
Hilina Worku ◽  
Debela Gemeda ◽  
...  
Keyword(s):  
Antimicrobial Susceptibility ◽  
Addis Ababa ◽  
Clinical Signs ◽  
Diagnosis And Treatment ◽  
Widal Test ◽  
Predictive Values ◽  
Blood Samples ◽  
Antimicrobial Susceptibility Tests ◽  
Stool Samples ◽  
Susceptibility Tests

Background. Diagnosis using reliable tools and treatment followingin vitroantimicrobial susceptibility tests are critical to proper addressing of antibiotic-resistantSalmonellainfection.Methodology. A cross-sectional study was conducted to assess the practice of diagnosis and treatment of salmonellosis in Addis Ababa. Tube Widal test (for blood samples only), culture, biochemical and carbohydrate fermentation, serotyping, and antimicrobial susceptibility tests were employed for both blood and stool samples.Results. Of all the diseases listed in the diagnosis, nontyphoidal (n=72, 13.71%) and typhoidal (n=47, 8.95%) salmonellosis were the second and third common diseases. Among the 288 blood samples, almost half were positive for O, H, or both antigens. However, only 1 (0.68%) of the positive blood samples yieldedSalmonellaisolate during culture. The study demonstrated low specificity (0.68%) and positive predictive value (48.78%) of Widal test. Conversely, the test showed 100% sensitivity and negative predictive values.Salmonellaisolates were identified from 7 (7.07%) of 99 stool samples. Two-thirds of salmonellosis suspected patients received antibiotic treatment. However, only half of the confirmed salmonellosis patients were treated with appropriate antibiotics. All of the isolates were susceptible to ciprofloxacin and ceftriaxone but resistant to ampicillin.Conclusions. Majority of the patients who participated in this study were wrongly diagnosed using symptoms, clinical signs, and tube Widal test. Consequently, most of the patients received inappropriate treatment.

scholarly journals Reliable Measurements of theβ-Amyloid Pool in Blood Could Help in the Early Diagnosis of AD

2012 ◽  
Vol 2012 ◽  
pp. 1-10 ◽  
Author(s):  
Pedro Pesini ◽  
Virginia Pérez-Grijalba ◽  
Inmaculada Monleón ◽  
Mercè Boada ◽  
Lluís Tárraga ◽  
...  
Keyword(s):  
Cognitive Impairment ◽  
Mild Cognitive Impairment ◽  
Early Diagnosis ◽  
Small Proportion ◽  
Blood Cells ◽  
Plasma Proteins ◽  
Blood Samples ◽  
Striking Result ◽  
Healthy Control ◽  
Amnesic Mild Cognitive Impairment

The present study was aimed at assessing the capability of Aβ1-40 and Aβ1-42 levels in undiluted plasma (UP), diluted plasma (DP), and cell bound (CB) to distinguish between early stages of Alzheimer's disease (AD), amnesic mild cognitive impairment (MCI), and healthy control (HC). Four blood samples from each participant were collected during one month and the levels of Aβ1-40 and Aβ1-42 were determined by a blinded proprietary ELISA sandwich (Araclon Biotech. Zaragoza, Spain). First striking result was that the amount of Aβ1-40 and Aβ1-42 in UP represented only a small proportion (~15%) of the total beta-amyloid pool in blood (βAPB) described here as the sum of Aβ1-40 and Aβ1-42 in blood where they are free in plasma, bound to plasma proteins, and bound to blood cells. Furthermore, we found that levels of Aβ1-40 and Aβ1-42 in UP, DP, and CB were significantly higher in MCI when compared to HC. On average, the totalβAPB was 1.8 times higher in MCI than in HC (P=0.03) and allowed to discriminate between MCI and HC with a sensitivity and specificity over 80%. Thus, quantification of several markers of theβAPB could be useful and reliable in the discrimination between MCI and HC.

scholarly journals Sequencing and structure analysis of UBC gene for breast and ‎lung cancer

2019 ◽  
Vol 10 (3) ◽  
pp. 1640-1645
Author(s):  
Saleen Salam Abdulhadi ◽  
Abbas Abdullah Mohammed‎
Keyword(s):  
Lung Cancer ◽  
Cancer Patients ◽  
Genetic Alterations ◽  
Cancer Disease ◽  
Blood Samples ◽  
Lung Cancer Patients ◽  
Control Subjects ◽  
Exon 1 ◽  
Healthy Control ◽  
Transition Mutation

In the present study, sequencing approach has been adopted for exploring the ‎genetic alteration of sequences for the ubiquitin gene (UBC) in patients of breast and ‎lung cancer and comparing the results with a normal sequence that obtained from NCBI. ‎The aim of this study was to detect for genetic alterations of UBC gene in the breast and ‎lung cancer patients then compare with healthy control subjects, to investigate the ‎association between the mutations at the intron region of the UBC gene and cancer disease, ‎‎40 blood samples were examined from patients with breast and lung cancer aged ranged from (17-65) years, were collected at Al-Amal Hospital of cancer in Baghdad ‎province/Iraq, the period of collecting samples were from October/2018 to January/2019. ‎While twenty-two blood samples from healthy control subjects were collected at ages ‎ranged from(19-59). After DNA extraction, the PCR primer was designed to amplify the ‎region in the UBC gene (part of exon 1 and the whole intron). Here we report the polymorphism of the intron sequence of the UBC gene in Iraqi population as the results of sequencing the PCR amplified products showed three different transition mutation G→A, ‎C→T, T→C in patients with breast cancer were also appeared in healthy control subjects. While nine transition mutations appeared in lung cancer patients, at different locations ‎of the sequence were detected by BLAST tool. ‎

scholarly journals Study of haematological, biochemical profile and clinical presentation in dengue positive patients: 82 cases

2018 ◽  
Vol 6 (6) ◽  
pp. 2099
Author(s):  
Mubin I. Patel ◽  
Abhishek Patel ◽  
Avani Patel ◽  
Sharmistha Patel ◽  
Suresh Padsala
Keyword(s):  
Dengue Fever ◽  
Clinical Presentation ◽  
Liver Enzymes ◽  
Neglected Tropical Diseases ◽  
Clinical Signs ◽  
Signs And Symptoms ◽  
Biochemical Profile ◽  
Igg Antibody ◽  
Aedes Mosquitoes ◽  
Clinical Signs And Symptoms

Background: Dengue Fever (DF) is a self-limiting disease caused by arbovirus and transmitted by Aedes mosquitoes (Aedes aegypti and Aedes albopictus). It is one of the 17 neglected tropical diseases by WHO. Diagnosis of dengue depends mainly on the detection of IgM and IgG antibody, and NS1 antigen.Methods: The study was carried out in Department of Pathology, affiliated with a government hospital. It includes 82 dengue patients, admitted from August 2015 to August 2016. Haematological, biochemical profile, clinical signs and symptoms were recorded. The Tourniquet test was performed in all the patients on admission. Grading of dengue: DF/DHFI/DHFII/DHFIII/DHFIV. Grade III and IV were collectively called as Dengue Shock Syndrome.Results: Total 82 Dengue positive cases were studied, 52 (63%) were males and 30 (37%) were females. 24 (29%) patients were recorded in September 22 (27%) in October 19 (23%) in August. 12 (14.60%) had positive tourniquet test. Thrombocytopenia was present in 86.5 % patients. Majority cases were of classical dengue fever 51 (62.20%), 14 (17.07%) were of DHF I, 12 (14.63%) were of DHF II, 3 (3.66%) were of DHF III and 2 (2.44%) were of DHF IV.Conclusions: It is very important to correlate clinical examination with haematological and biochemical profile in dengue patients. Hematocrit value, leucopenia, thrombocytopenia, raised liver enzymes is very important to monitor dengue cases in their initial stages and thus facilitate early treatment. This would minimize morbidity and mortality arising out of serious complications of dengue fever.

scholarly journals Detection of Aspergillus species in bone marrow transplant patients

2010 ◽  
Vol 4 (08) ◽  
pp. 511-516 ◽  
Author(s):  
Parisa Badiee ◽  
Abdolvahab Alborzi
Keyword(s):  
Bone Marrow ◽  
Real Time ◽  
Bone Marrow Transplant ◽  
Real Time Pcr ◽  
Clinical Signs ◽  
Marrow Transplant ◽  
Aspergillus Species ◽  
Pcr Assay ◽  
Blood Samples ◽  
Transplant Patients

Introduction:  Invasive aspergillosis is a severe complication of cytotoxic chemotherapies and bone marrow transplantation (BMT). The aim of this study was to assess the utility of a real-time PCR assay for the early diagnosis of Aspergillus species in blood samples from BMT patients. Methodology: Blood specimens (n = 993) from patients (n = 82) scheduled for BMT were collected prior to transplant and for 100 days post transplantation.  The specimens were later tested using an Aspergillus-specific real-time PCR assay. Cultures of clinical samples, along with sonography and computerized tomographic scans, were performed as standard of care. Results: Aspergillus DNA was positive in 94 sequential blood samples from 13 patients with clinical and radiological signs of infection. Samples from three of these patients were PCR-positive for Aspergillus in the first week of admission, prior to transplantation. Four patients with aspergillosis were cured with antifungal agents and nine died. An additional 12 patients without clinical signs of infection were PCR-positive on one occasion each, while two patients with clinical signs of infection were PCR-negative. Compared to routine methods of aspergillosis diagnosis, the respective sensitivity, specificity, negative, and positive predictive values of the PCR method by patient were 86.6%, 82%, 96.5% and 52%. Conclusions: The results show that Aspergillus infections in the blood of bone marrow transplant patients can be dectected by PCR methods. Early detection of Aspergillus infections by PCR has the potential to positively impact patient mortality rate and provide cost savings to hospitals.

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