scholarly journals Complete diagnosis of leptospirosis in tropical reproductive cattle

2018 ◽  
Author(s):  
Gabriela Pacheco Sánchez ◽  
Fabio Almeida de Lemos ◽  
Mirian Dos Santos Paixão-Marques ◽  
Maria Fernanda Alves-Martin ◽  
Lívia Maísa Guiraldin ◽  
...  
Keyword(s):  
Farm Workers ◽  
Diagnostic Methods ◽  
Economic Losses ◽  
Reproductive Disorders ◽  
Semisolid Medium ◽  
Sanitary Condition ◽  
Prophylactic Measures ◽  
Polymerase Chain ◽  
Economic Balance ◽  
Sanitary Conditions

AbstractLeptospirosis is a worlwide zoonosis of great impact in both animal and public health. Bovine leptospirosis is commonly manifested by reproductive disorders, such as abortion, stillbirth and infertility; causing depletion of the economic balance of livestock farms, along with representing a health risk problem for farm workers. In view of these consequences, we aimed to evaluate the sanitary status of tropical cattle and their role as reproductive disseminators of leptospirosis. We analyzed blood and semen samples from 11 brazilian herds by three diagnostic methods -Culture, Microscopic Agglutination Test and Polymerase Chain Reaction. All animals were negative for bacteriological culture in Fletcher’s semisolid medium; 66% (264/400) animals were seropositive to at least one of 19 serovars (17 serogroups) ofLeptospiraspp. by MAT, given that 42.4% and 5.3% of animals presented titers against brazilians isolates Guaricura and Nupezo, respectively; furthermore, five animals were positive by PCR in blood and/or culture samples and three semen samples were positive by PCR (one of them also seropositive). These results highlight the coexistence of both disease's stages (acute and chronic) in the same environment, thus alert for venereal dissemination of leptospirosis, aggravating their sanitary condition and fomenting economic losses. We, authors, recommend the adoption of prophylactic measures, such as systemic vaccination, treatment of animals and improvement of hygienic-sanitary conditions.

scholarly journals HoBi-Like Pestivirus and Reproductive Disorders

2020 ◽  
Vol 7 ◽  
Author(s):  
Nicola Decaro
Keyword(s):  
Reproductive Tract ◽  
Diagnostic Methods ◽  
Economic Losses ◽  
Reproductive Disorders ◽  
Fatal Disease ◽  
Viral Diarrhea ◽  
Asymptomatic Infections ◽  
Cattle Reproduction ◽  
The Impact

HoBi-like pestivirus (HoBiPeV) is an emerging group of pestiviruses that has been detected in cattle and other ruminants in South America, Europe, and Asia. Analogous to other bovine pestiviruses, namely bovine viral diarrhea (BVDV) 1 and 2, HoBiPeV is able to cause a variety of clinical forms that range from asymptomatic infections to fatal disease, having a great impact on cattle productions and causing substantial economic losses, mainly as a consequence of the occurrence of reproductive failures. The manuscript aims to provide an updated review of the currently available literature about the impact of HoBiPeV infection on cattle reproduction. The reproductive disorders observed in cattle due to natural and experimental infections caused by this virus are reported along with the few available in-vitro studies involving the reproductive tract. HoBiPeV should be considered among the bovine pathogens that impact on reproduction, but there is a need for more specific and sensitive diagnostic methods, while the cross-protection elicited by commercially available BVDV vaccines should be better investigated.

scholarly journals Genetic and parasitological identification of Trypanosoma evansi infecting cattle in South Sulawesi, Indonesia

2021 ◽  
Vol 14 (1) ◽  
pp. 113-119
Author(s):  
Agus Setiawan ◽  
Wisnu Nurcahyo ◽  
Dwi Priyowidodo ◽  
Rina Tri Budiati ◽  
Desy Sylvia Ratna Susanti
Keyword(s):  
Clinical Signs ◽  
Trypanosoma Evansi ◽  
Buffy Coat ◽  
Economic Losses ◽  
Pcr Analysis ◽  
Its2 Region ◽  
Reproductive Disorders ◽  
South Sulawesi ◽  
Rdna Sequencing ◽  
Polymerase Chain

Background and Aim: Sulawesi is an Indonesian island located within the Wallacea region that contains a distinctive mix of Asian and Australasian species. This distinctiveness extends to parasites, including Trypanosoma evansi, the cause of surra. Surra has non-specific clinical signs such as anemia, anorexia, weight loss, drop in milk production, and reproductive disorders which cause economic losses. Due to the trade of livestock, surra has spread in Indonesia from one island to another. The aim of this study was to investigate the trypanosomes infecting cattle in South Sulawesi, using internal transcribed spacer (ITS2) ribosomal DNA (rDNA) sequencing. Materials and Methods: A total of 100 whole blood samples were collected from cattle in Makassar, South Sulawesi Province, Indonesia. All samples were tested using conventional parasitological methods (CPT), namely, thin blood smear, buffy coat smears, and polymerase chain reaction (PCR) testing. Positive PCR results were sequenced and phylogenetically analyzed. Results: Only one of the 100 samples was found to be positive with microscopic observation; however, PCR analysis revealed that 3% (3/100) of samples were positive. Sequencing identified the positive samples as T. evansi, China isolate (KU552344), with a homology of 99%. Two out of three sequences showed variations in ITS2 region. Conclusion: Based on CPT and molecular analysis, T. evansi isolates from infected cattle in South Sulawesi demonstrate genetic diversity of ITS2 sequences.

Recent advances in nucleic acid-based methods for detection of helminth infections and the perspective of biosensors for future development

Parasitology ◽  
2019 ◽  
Vol 147 (4) ◽  
pp. 383-392 ◽  
Author(s):  
Hanif Ullah ◽  
Abdul Qadeer ◽  
Muhammad Rashid ◽  
Muhammad Imran Rashid ◽  
Guofeng Cheng
Keyword(s):  
Nucleic Acid ◽  
Helminth Infection ◽  
Diagnostic Methods ◽  
Economic Losses ◽  
Chain Reaction ◽  
Advantages And Disadvantages ◽  
Recent Advances ◽  
Helminth Infections ◽  
Pathogen Control ◽  
Polymerase Chain

AbstractPathogenic helminth infections are responsible for severe health problems and economic losses worldwide. Timely and accurate diagnosis of helminth infections is critical for adopting suitable strategies for pathogen control. Here, we review recent advances in nucleic acid-based diagnostic methods, including polymerase chain reaction, quantitative qPCR, loop-mediated isothermal amplification and recombinase polymerase amplification, and discuss their advantages and disadvantages for diagnosing helminth infections. In addition, we highlight recent advances in biosensors for the detection of nucleic acid biomarkers that can potentially be used for the diagnosis of helminth infection.

scholarly journals Strengthening of Molecular Diagnosis of SARS-CoV-2 / COVID-19 with a Special Focus on India

2020 ◽  
Vol 14 (suppl 1) ◽  
pp. 789-798
Author(s):  
Ragini Bhatia ◽  
Rajesh Chaudhary ◽  
Sandip Kumar Khurana ◽  
Ruchi Tiwari ◽  
Kuldeep Dhama ◽  
...  
Keyword(s):  
Laboratory Testing ◽  
Nasal Congestion ◽  
Diagnostic Methods ◽  
Economic Losses ◽  
Clinical Samples ◽  
Special Focus ◽  
Runny Nose ◽  
Public Health Professionals ◽  
Polymerase Chain ◽  
Novel Coronavirus

Severe acute respiratory syndrome corona virus-2 (SARS-CoV-2), a novel coronavirus initially reported in Wuhan, China, is the causative agent of coronavirus disease (COVID-19) pandemic. Symptoms of the disease comprise of fever, tiredness, dry cough, aches and pains, nasal congestion, runny nose, sore throat, diarrhoea and pneumonia at the late stage. SARS-CoV-2 has severely crippled the healthcare system and has caused huge economic losses. Following the outbreak, the SARS-CoV-2 was recognized timely and its genome was sequenced, leading to the development of real-time polymerase chain reaction assays for its detection in clinical samples collected from suspected cases. The management of the pandemic is limited by a number of misconceptions and insufficient information about laboratory testing for SARS-CoV-2 to confirm the disease. This includes a lack of awareness about procedures for the collection, transport, testing, and handling of biological samples for COVID diagnosis. This article provides an overview of the current laboratory diagnostic methods with a purpose to provide information and guidance to laboratories, stakeholders, broader community and especially public health professionals involved in laboratory testing for SARS-CoV-2.

scholarly journals High Frequency of Cryptosporidium hominis Infecting Infants Points to A Potential Anthroponotic Transmission in Maputo, Mozambique

Pathogens ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 293
Author(s):  
Idalécia Cossa-Moiane ◽  
Hermínio Cossa ◽  
Adilson Fernando Loforte Bauhofer ◽  
Jorfélia Chilaúle ◽  
Esperança Lourenço Guimarães ◽  
...  
Keyword(s):  
Public Hospitals ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
P Value ◽  
Cryptosporidium Hominis ◽  
Cryptosporidium Spp ◽  
Pcr Rflp ◽  
Maputo City ◽  
Stool Samples ◽  
Polymerase Chain

Cryptosporidium is one of the most important causes of diarrhea in children less than 2 years of age. In this study, we report the frequency, risk factors and species of Cryptosporidium detected by molecular diagnostic methods in children admitted to two public hospitals in Maputo City, Mozambique. We studied 319 patients under the age of five years who were admitted due to diarrhea between April 2015 and February 2016. Single stool samples were examined for the presence of Cryptosporidium spp. oocysts, microscopically by using a Modified Ziehl–Neelsen (mZN) staining method and by using Polymerase Chain Reaction and Restriction Fragment Length Polymorphism (PCR-RFLP) technique using 18S ribosomal RNA gene as a target. Overall, 57.7% (184/319) were males, the median age (Interquartile range, IQR) was 11.0 (7–15) months. Cryptosporidium spp. oocysts were detected in 11.0% (35/319) by microscopy and in 35.4% (68/192) using PCR-RFLP. The most affected age group were children older than two years, [adjusted odds ratio (aOR): 5.861; 95% confidence interval (CI): 1.532–22.417; p-value < 0.05]. Children with illiterate caregivers had higher risk of infection (aOR: 1.688; 95% CI: 1.001–2.845; p-value < 0.05). An anthroponotic species C. hominis was found in 93.0% (27/29) of samples. Our findings demonstrated that cryptosporidiosis in children with diarrhea might be caused by anthroponomic transmission.

Comparison of Droplet Digital Polymerase Chain Reaction (ddPCR) and Real-Time Quantitative Polymerase Chain Reaction (qPCR) in Detecting Neonatal Invasive Fungal Infections

2021 ◽  
Vol 11 (3) ◽  
pp. 373-379
Author(s):  
Huitao Li ◽  
Xueyu Chen ◽  
Xiaomei Qiu ◽  
Weimin Huang ◽  
Chuanzhong Yang
Keyword(s):  
Polymerase Chain Reaction ◽  
Limit Of Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Diagnostic Methods ◽  
Chain Reaction ◽  
Fungal Isolation ◽  
Blood Samples ◽  
Fungal Strains ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction

Invasive fungal infection (IFI) is the leading cause of death in neonatal patients, yet the diagnosis of IFI remains a major challenge. At present, most IFI laboratory diagnostic methods are based on classical, but limited, methods such as fungal isolation and culture and histopathological examination. Recently, quantitative polymerase chain reaction (qPCR) and droplet digital polymerase chain reaction (ddPCR) technology have been adopted to quantify nucleic-acid identification. In this study, we established qPCR and ddPCR assays for IFI diagnosis and quantification. qPCR and ddPCR were carried out using identical primers and probe for the amplification of 18S rRNA. Assay results for three fungal strains were positive, whereas ten non-fungal strains had negative results, indicating 100% specificity for both ddPCR and qPCR methods. Genomic DNA of Candida albicans was tested after a serial dilution to compare the sensitivity of the two PCR methods. The limit of detection of ddPCR was 3.2 copies/L, which was a ten-fold increase compared with that of the qPCR method (32 copies/L). Blood samples from 127 patients with high-risk factors and clinical symptoms for IFI were collected from a NICU in Shenzhen, China, and analyzed using qPCR and ddPCR. Thirty-four blood samples from neonates had a proven or probable diagnosis of IFI, and 25 of these were positive by qPCR, whereas 30 were positive by ddPCR. Among the 93 blood samples from neonates who had a possible IFI or no IFI, 24 were positive using qPCR, and 7 were positive using ddPCR. In conclusion, ddPCR is a rapid and accurate pan-fungal detection method and provides a promising prospect for IFI clinical screening.

Studies on Diagnosis of Bovine Brucellosis by Immunocyto-chemistry and Immunohistochemistry

2021 ◽  
Author(s):  
K.S. Lakshmikanth ◽  
N.S. Sharma ◽  
D. Pathak ◽  
Paviter Kaur
Keyword(s):  
Biochemical Analysis ◽  
Placental Tissue ◽  
Diagnostic Techniques ◽  
Clinical Samples ◽  
Bovine Brucellosis ◽  
Reproductive Disorders ◽  
Chain Reaction ◽  
Tissue Samples ◽  
Polymerase Chain ◽  
Positive Results

Background: Brucellosis is a major threat to livestock economy and an important zoonotic disease. A rapid and accurate diagnosis is a necessity to curb the spread and progress of the disease. The current study aimed to evaluate sensitivity of Immunocytochemistry and Immunohistochemistry methods for detection of Brucella spp.Methods: A total of 50 samples comprising of fetal stomach content, vaginal discharges and placenta were collected from cattle and buffaloes suffering from abortions and other reproductive disorders in and around Ludhiana, Punjab during the period 2017-2018. All the samples were processed for isolation and confirmed with biochemical analysis and Polymerase chain reaction (PCR). The isolates obtained and 43 clinical samples excluding placental samples were subjected to Immunocytochemistry (ICC). Immunohistochemistry (ICH) was performed on placental samples.Result: A total of four isolates were recovered from the screened samples. The four isolates also yielded positive results in Immunocytochemistry. Among the 43 clinical samples screened by Immunocytochemistry, five were positive, however only 3 isolates were recovered on isolation. A total of seven placental tissue samples were processed and subjected to immunohistochemistry. Of the three placental samples positive by immunohistochemistry, only one sample was isolated on culture. The results suggest that both immunocytochemistry and immunohistochemistry are sensitive diagnostic techniques in comparison to isolation.

scholarly journals UTILITY OF ANTIGEN DETECTION TEST AND POLYMERASE CHAIN REACTION IN THE DIFFERENTIATION OF TUBERCULOUS AND NON-TUBERCULOUS MYCOBACTERIA

2017 ◽  
Vol 10 (6) ◽  
pp. 289
Author(s):  
Yogita Singh ◽  
Raji Vasanth ◽  
Shrikala Baliga ◽  
Dhanashree B
Keyword(s):  
Polymerase Chain Reaction ◽  
False Negative ◽  
Diagnostic Methods ◽  
Clinical Samples ◽  
Chain Reaction ◽  
College Hospital ◽  
Medical College ◽  
Negative Results ◽  
Polymerase Chain ◽  
Mycobacterium Spp

Objectives: Cultivation and identification of mycobacteria to species level remains difficult and time-consuming. Hence, easy and rapid diagnostic methods are necessary for the differentiation of Mycobacterium tuberculosis (MTB) from non-tuberculous mycobacteria (NTM). The present study aims to detect and differentiate MTB from NTM isolated from clinical samples by immunochromatographic test (ICT) and polymerase chain reaction (PCR). Methods: Over a period of 1 year, clinical samples (n=496) received from suspected cases of TB, at the Department of Microbiology, Kasturba Medical College Hospital, Mangalore were cultured to isolate Mycobacterium spp. Identification of all the isolates was done by conventional biochemical technique, ICT, and PCR. Results: Among the 496 samples processed, 49 (9.87%) were acid-fast bacilli smear positive and 59 (11.89%) samples showed the growth of Mycobacterium spp. Among these, 10 were rapid growers, 49 were slow-growing mycobacteria, out of which 30 were MTB as identified by conventional biochemical reaction. Out of 59 Mycobacterial isolates subjected to ICT for the detection of MPT 64 antigen, only 28 were identified as MTB. However, all the 30 isolates were correctly identified as MTB by PCR. Conclusion: Hence, PCR is essential for rapid differentiation of non-tuberculous Mycobacterium from MTB. False negative results seen with immunochromatographic MPT 64 antigen assay could be due to mutations within the mpt64 gene. Further studies are necessary to characterize these PCR-positive and immunochromatographic assay negative MTB isolates.

scholarly journals Comparative Study of Immune-Diagnostic Tools with Polymerase Chain Reaction in Sub-Clinical Leishmaniasis Isolates

2011 ◽  
Vol 12 (1) ◽  
pp. 34-39 ◽  
Author(s):  
Nazar M Abdalla
Keyword(s):  
Polymerase Chain Reaction ◽  
Skin Test ◽  
Diagnostic Methods ◽  
Diagnostic Tools ◽  
Enzyme Linked Immunosorbent Assay ◽  
Clinical Form ◽  
Chain Reaction ◽  
Leishmanin Skin Test ◽  
Wide Range ◽  
Polymerase Chain

Objective: This study aimed to identify cases of leishmaniasis in the Nuba Mountain area, which is situated in a unique geographical site located in the centre of Sudanese leishmania belt. Wide range of investigations are available for detection of leishmania cases, but still the most reliable and easy test used as screening and epidemiological tool in field studies needs to be evaluated. The most commonly used conventional diagnostic methods direct microscopy and culture have some drawbacks in diagnosing subclinical cases of leishmaniasis. Materials and methods: In this study, comparative properties of various immune-diagnostic tools with Polymerase Chain Reaction used in sub-clinical leishmaniasis isolates were explored. The immune-diagnostic tools involved in this study include- Leishmanin Skin Test (LST), Enzyme Linked Immunosorbent Assay (ELISA) and Direct Agglutination Test (DAT). The study was conducted in the Green Valley village (Rashad Province, South Kordofan State) with a population of 332. Most of the villagers presented with sub-clinical form of leishmaniasis with minor symptoms and signs without the features found in clinical form of visceral leishmaniasis such as fever, diarrhoea, epistaxis, enlarged lymph nodes, spleen and liver. In this study we collected demographic, clinical and epidemiological data using special questionnaire. Leishmanin skin test (LST), ELISA, DAT and PCR for parasite DNA detection were used. Result: The final positive cases detected by PCR were 32 out of 332 belong to L. donovani species. The final positive cases detected by LST were 51.2% of the total population under study, while 11 out of the 37 tested samples were positive by ELISA. All of the 332 villagers showed negative readings by DAT with exception of three individuals who were positive with very high titers. Conclusion: DNA etxtraction and amplification with primers can be a good screening tool in subclinical leishmaniasis isolates. Keyword: Sub-clinical; Leishmaniasis; Leishmanin Skin Test; ELISA; DAT; PCR. DOI: 10.3329/jom.v12i1.5422J Medicine 2011; 12 : 34-39

Simultaneous detection of astrovirus, rotavirus, reovirus and adenovirus type I in broiler chicken flocks

2012 ◽  
Vol 15 (2) ◽  
pp. 337-344 ◽  
Author(s):  
D. Roussan ◽  
I. Shaheen ◽  
G. Khawaldeh ◽  
W. Totanji ◽  
R. Al-Rifai
Keyword(s):  
Polymerase Chain Reaction ◽  
Broiler Chicken ◽  
Simultaneous Detection ◽  
Adenovirus Type ◽  
Economic Losses ◽  
Type I ◽  
Chain Reaction ◽  
Group I ◽  
Avian Nephritis Virus ◽  
Polymerase Chain

Simultaneous detection of astrovirus, rotavirus, reovirus and adenovirus type I in broiler chicken flocksEnteric diseases cause substantial economic losses to the poultry industry. Astroviruses, rotaviruses, reoviruses, and adenovirus type 1 have been reported as a significant cause of intestinal symptoms in poultry. In the present study, intestinal samples from 70 commercial broiler chicken flocks were examined for the presence of astroviruses, rotavirus, and reovirus by reverse transcription-polymerase chain reaction, and for the presence of group I adenovirus by polymerase chain reaction. Astroviruses were identified in 38.6% of samples tested. Both avian nephritis virus and chicken astrovirus were identified in the astrovirus positive flocks, where 74.1% of these flocks were positive for only one type of astrovirus, whereas, 25.9% of these flocks were positive for both types of astrovirus. Reoviruses, rotaviruses, and adenoviruses were identified in 21.4, 18.6, and 14.3% of these flocks, respectively. Concomitant infection with two or more viruses in the same flock were also prominent, where 5.7, 5.7, 2.9, 2.9, 1.4, and 1.4% of these flocks were positive with both astrovirus and rotavirus; astrovirus and adenovirus; astrovirus and reovirus; rotavirus and adenovirus; rotavirus and reovirus; and reovirus and adenovirus respectively. Moreover, 4.3 and 2.7% of these flocks were positive for astrovirus, reovirus, and adenovirus; and astrovirus, reovirus, and rotavirus, respectively. Further studies will focus on identifying specific viral factors or subtypes/subgroups associated with disease through pathogenesis studies, economic losses caused by infections and co-infections of these pathogens, and the costs and benefits of countermeasures.

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