Cystic Echinococcosis In Sheep Research Articles

Determination of Malondialdehyde, Nitric Oxide, Reduced Glutathione, Sialic Acid and Ceruloplasmin Levels in Sheep Liver Tissue With Hydatic Cyst

Cystic echinococcosis is a prevalent helminth-zoonosis that poses a significant threat to human and animal health worldwide. Although it typically shows an asymptomatic clinical course, it has been reported to cause many damages and biochemical changes in tissues and organism. The aim of this study was to investigate the concentrations of nitric oxide (NO), reduced glutathione (GSH), malondialdehyde (MDA), ceruloplasmin (Cp), and total sialic acid (TSA) in sheep liver tissue affected by cystic echinococcosis. The study was carried out on sheep between 4-5 years of age brought to the slaughterhouse in the Igdir territory. The livers were examined post-mortem for cystic echinococcosis and cystic structures. Sheep liver tissues that tested positive for protoscolex were designated as the ‘infected group’, while healthy sheep liver tissues without lesions were assigned to the ‘control group’. The results showed significantly higher levels of NO, MDA, Cp and TSA in the infected group compared to the control group (p<0.05), while GSH levels were significantly lower (p<0.05). These findings indicate that cystic echinococcosis in sheep is closely associated with mechanisms of inflammation, oxidative stress, and tissue damage. Moreover, our study provides insights into the oxidative response of cystic echinococcosis in liver tissue and enhances our understanding of the disease’s pathogenesis.

Rapid and non-invasive detection of cystic echinococcosis in sheep based on serum fluorescence spectrum combined with machine learning algorithms.

Cystic echinococcosis (CE) is a grievous zoonotic parasitic disease. Currently, the traditional technology of screening CE is laborious and expensive, developing an innovative technology is urgent. In this study, we combined serum fluorescence spectroscopy with machine learning algorithms to develop an innovative screening technique to diagnose CE in sheep. Serum fluorescence spectra of Echinococcus granulosus sensu stricto-infected group (n = 63) and uninfected E. granulosus s.s. group (n = 60) under excitation at 405 nm were recorded. The linear support vector machine (Linear SVM), Quadratic SVM, medium radial basis function (RBF) SVM, K-nearest neighbor (KNN), and principal component analysis-linear discriminant analysis (PCA-LDA) were used to analyze the spectra data. The results showed that Quadratic SVM had the great classification capacity, its sensitivity, specificity, and accuracy were 85.0%, 93.8%, and 88.9%, respectively. In short, serum fluorescence spectroscopy combined with Quadratic SVM algorithm has great potential in the innovative diagnosis of CE in sheep.

Immunohistochemical Alterations of Syndecan-1 in Sheep Liver with Cystic Echinococcosis

Cystic echinococcosis is formed by the larval forms of Echinococcus granulosus, causes health problems in humans and different species of animals worldwide. There is still limited information about the pathophysiological response to cystic echinococcosis in the liver. Syndecan-1 (Sdc1) is a cell surface proteoglycan of the liver. It has a crucial role in the pathophysiology of various liver diseases. This study aimed to investigate the immunohistochemical expression of Sdc1 in the liver with cystic echinococcosis in sheep. A total number of 51 liver tissue samples with cystic echinococcosis and ten healthy livers were examined. The tissue samples were stained with hematoxylin-eosin (HE) for histopathological examinations. Sdc1 was determined in the same liver tissues by immunohistochemistry. Infected liver tissues mainly showed severe inflammatory reactions, congestion, diffuse degeneration, and necrosis of hepatocytes around the cysts. The results indicated severe Sdc1-positive staining in hepatocytes around the cysts. Liver tissues of the control group showed relatively mild immunopositive reactions. This study determined that the Sdc1 significantly increased in the sheep livers with cystic echinococcosis infection.

Echinococcosis of animals and humans as a socially dangerous problem in a densely populated subject of the North Caucasus

Echinococcosis of animals and humans as a socially dangerous problem in a densely populated subject of the North Caucasus

Rapid and accurate screening of cystic echinococcosis in sheep based on serum Fourier-transform infrared spectroscopy combined with machine learning algorithms.

Cystic echinococcosis (CE) in sheep is a serious zoonotic parasitic disease caused by Echinococcus granulosus sensu stricto (s.s.). Presently, the screening technology for CE in sheep is time-consuming and inaccurate, and novel screening technology is urgently needed. In this work, we combined machine-learning algorithms with Fourier transform infrared (FT-IR) spectroscopy of serum to establish a quick and accurate screening approach for CE in sheep. Serum samples from 77 E. granulosus s.s.-infected sheep to 121 healthy control sheep were measured by FT-IR spectrometer. To optimize the classification accuracy of the serum FI-TR method for the E. granulosus s.s.-infected sheep and healthy control sheep, principal component analysis (PCA), linear discriminant analysis, and support vector machine (SVM) algorithms were used to analyze the data. Among all the bands, 1500-1700 cm-1 band has the best classification effect; its diagnostic sensitivity, specificity, and accuracy of PCA-SVM were 100%, 95.74%, and 96.66%, respectively. The study showed that serum FT-IR spectroscopy combined with machine learning algorithms has great potential for rapid and accurate screening methods for the CE in sheep.

The Diagnosis of Sheep Cystic Echinococcosis by Native Antigen B - Elisa Method

Cystic echinococcosis (CE) is one of the most prevalent zoonotic diseases in some countries in the world. Cystic echinococcosis is considered a neglected disease. This disease increases economic damage via medical costs and loss of human and livestock productivity. The aim of the current study gains a better understanding of the prevalence of CE in sheep. Totally 250 sheep sera were collected. 25 serum samples from newborn lambs were used as negative, and 25 others were obtained from slaughterhouses as positive to infection with CE and 200 unknown sera. Antigen B Isolated from hydatid cysts fluid was used for designing ELISA methods. The first Method used anti-Sheep conjugate (SIGMA, at 1:3000 dilution). According to the results, the seroprevalence of CE in East Azerbaijan of Iran was 5.5% and sensitivity and specificity for the diagnosis of hydatidosis in sheep by AgB-ELISA methods was 92%. Using Antigen B in ELISA design for hydatidosis diagnosis has attracted researchers in recent years. During this study, an Iranian native B antigen was used to design the specific detection of hydatidosis in sheep using a specific ELISA technique. The results have shown that using Antigen B in ELISA design is so valuable.

Protection against cystic echinococcosis in sheep using an Escherichia coli-expressed recombinant antigen (EG95) as a bacterin.

The EG95 recombinant vaccine is protective against cystic echinococcus in animal intermediate hosts. Preparation of the existing, registered EG95 vaccines involves semi-purification of the vaccine protein, adding to the cost of production. Truncation of the EG95 cDNA, shortening both the amino and carboxy-termini of the protein, leads to high levels of recombinant protein expression. The recombinant EG95 protein was prepared as a bacterin from clarified, whole bacterial lysate, and used in a vaccine trial in sheep against an experimental challenge infection with Echinococcus granulosus eggs. The EG95 bacterin was found to induce 98% protection. Use of this in a new generation EG95 vaccine would simplify production, facilitate new sources of the vaccine and potentially enhance uptake of vaccination in control of E. granulosus transmission.

The Diagnosis of Sheep Cystic Echinococcosis by Native Antigen B - Elisa Method

Cystic echinococcosis (CE) is one of the most prevalent zoonotic diseases in some countries in the world. Cystic echinococcosis is considered a neglected disease. This disease increases economic damage via medical costs and loss of human and livestock productivity. The aim of the current study gains a better understanding of the prevalence of CE in sheep. Totally 250 sheep sera were collected. 25 serum samples from newborn lambs were used as negative, and 25 others were obtained from slaughterhouses as positive to infection with CE and 200 unknown sera. Antigen B Isolated from hydatid cysts fluid was used for designing ELISA methods. The first Method used anti-Sheep conjugate (SIGMA, at 1:3000 dilution). According to the results, the seroprevalence of CE in East Azerbaijan of Iran was 5.5% and sensitivity and specificity for the diagnosis of hydatidosis in sheep by AgB-ELISA methods was 92%. Using Antigen B in ELISA design for hydatidosis diagnosis has attracted researchers in recent years. During this study, an Iranian native B antigen was used to design the specific detection of hydatidosis in sheep using a specific ELISA technique. The results have shown that using Antigen B in ELISA design is so valuable.

Cystic echinococcosis in slaughtered sheep in Erzurum province, Turkey

Cystic echinococcosis (CE) is an important zoonotic infection caused by the larval stages of the genus Echinococcus. Turkey is a highly endemic region for CE and the disease is one of the major public health problems. The study was aimed to assess the situation of the CE in sheep in Turkey and also to provide data on circulating genotypes in the country. A total of 3319 sheep at slaughter were screened during the study. The prevalence of CE in the study area was 31.7% (1052/3319). The lungs were the most frequently CE infected organ (50%, 526/1052). Microscopic examination revealed that overall cyst fertility was 68.1%. Molecular analysis of partial fragments of 12S and COI gene regions were included for 351 selected cyst samples and all of them were identified as E. granulosus sensu stricto. Sequence analysis showed that the predominant genotype in the study areas was G1 (77.1%), and the rest were G3 (22.9%). The prevalence rate of CE in sheep in the study area is lower compared to previous years except for one province. Considering the high cyst fertility rate and the predominance of E. granulosus G1 which is particularly pathogenic to humans, calls for serious control measures like public awareness about the disease, sufficient dog deworming programs, continuity of monitoring the disease should be taken.

Assessment of the Vaccination Program against Cystic Echinococcosis in Sheep in the Pehuenche Community of Central Chile.

Echinococcosis is a neglected zoonosis that uses dogs and sheep as its main hosts in Chile. The Eg95 vaccine against sheep infection has been included in some control programs. Here, we assess the efficacy of the vaccination program in the hyperendemic Alto Biobío commune after 3 years of execution. Fisher's test and generalized linear models were used in the assessment. The program tried to offer a first dose at 2 months of age, a booster 1 month later, and yearly vaccination. Given logistic difficulties, important delays in vaccination occurred, and most animals did not receive the first booster. Dog deworming was not included in the program. Likely due to the aforementioned factors, the overall frequency of infection was not lower, but the proportion of large (>5 mm) cysts and fertile cysts was smaller after the program. The frequency of infection and/or the number of cysts were lower when the age at first dose was younger and the first booster was administered 1 month after the first dose. The results suggest that vaccination affects both cyst development after the larvae reach the target organs, as well as the development of the protoscolex once the cysts start developing.

Current Situation of Cystic Echinococcosis in Sheep in Turkey

Current Situation of Cystic Echinococcosis in Sheep in Turkey

Proteomic characterization of Echinococcus granulosus sensu stricto, Taenia hydatigena and Taenia multiceps metacestode cyst fluids

Cystic echinococcosis (CE) diagnosis by means of serological assays is hampered by the presence of parasites closely related to Echinococcus granulosus sensu lato (s.l.), responsible of the zoonotic disease and with which share cross-reacting antigens. Thus, improvements on the characterization of Echinococcus specific antigens expressed in the larval stage are required, in order to provide useful information for the development of immunological assays for the serodiagnosis of CE in sheep. Here, the proteome of the hydatid cyst fluids (HFs) of Echinococcus granulosus (hydatid fluid, EgHF) and other ovine parasites cyst fluids (CFs), Taenia hydatigena (ThCF) and Taenia multiceps (TmCF) were analyzed by a shotgun proteomic approach. Parasite and host protein profiles in the three types of cyst fluids were characterized and compared. Among the identified proteins, differential parasitic markers with serodiagnostic potential, due to their well-known immunoreactivity in human, included Ag5, AgB proteins, 8-kDa glycoproteins, hydatid disease diagnostic antigen P29 and major egg antigen P40. In particular, seven proteoforms of AgB and 8-kDa glycoprotein resulted to be the most promising diagnostic biomarkers, as they might predict CE in ovine and discriminate between different types of parasites.

Cystic echinococcosis in sheep and goats of Lebanon.

Cystic echinococcosis (CE), a zoonotic disease caused by the species complex of Echinococcus granulosus sensu lato (s.l.), is endemic in Middle East and the Mediterranean basin, where pastoral activity is widespread. Despite the chronic endemicity of the disease in Lebanon and neighbouring countries, recent data are scant. The objectives of this survey were to evaluate the current epidemiology of CE in Lebanon, investigate the prevalence and determine the infecting genotypes in locally raised sheep and goats. A multidimensional approach combining post-mortem inspection of slaughtered animals and molecular diagnosis of the parasite was conducted to this end. From 2018 to 2020, 62.9% of the sheep and 20.9% of the goats were found positive for CE. The presence of hydatids varied between organs, showing higher prevalence in the liver of sheep vs the lungs of goats, however, a higher fertility rate of hydatid cyst was observed in lungs for both animals. Molecular diagnosis based on partial mitochondrial cox1 gene (795 bp) showed that the majority of isolates were identified as E. granulosus sensu stricto (98.7%) and only one isolate from goat was identified as Echinococcus canadensis (genotype G7; 1.3%). Echinococcus granulosus s.s. population among the sheep and goats was represented by 22 haplotypes having very little genetic differentiation and relatively moderate haplotype diversity. Population demographics explored through neutrality indices suggested expanding population within the intermediate hosts. These results document the high prevalence of CE in the livestock of Lebanon and reveal for the first time the presence of three different genotypes G1, G3 and G7.

Fast Focus Ultrasound Liver Technique for the Assessment of Cystic Echinococcosis in Sheep.

Simple SummaryCystic echinococcosis (CE) is a parasitic zoonosis caused by the larval stage of the tapeworm Echinococcus granulosus. The liver ultrasonography can be considered as the gold standard for the in vivo diagnosis detection of CE. Nevertheless, control programs against E. granulosus are considered long-term actions that require an integrated approach and the high expenditure of time and financial resources. This parasitosis still causes severe economic losses both in human healthcare and in the entire livestock breeding system, with loss in milk, meat, and wool production. Considering the impact of this zoonosis in the modern sheep’s breeding system, a reliable hepatic ultrasound could represent an innovative strategy to control and even eradicate the disease in sheep farms. Based on these considerations, the present study aimed to evaluate a fast-focused technique for hepatic CE detection in different sheep breeds and compare the performance of the latter with another fast-scan (previously evaluated in Sarda sheep breed), the complete ultrasound liver examination, and the anatomopathological examination.A complete ultrasound examination (cUS) of the liver was performed on 172 female sheep and compared to the performance of a fast-focused ultrasound technique to diagnose echinococcal cysts. The scanned area was divided in: HYP (right hypocondrium), zone (Z)1 from HYP to the 11th intercostal space (IS), Z2 (10th–8th IS) and Z3 (7th–5th IS). Contiguous scans were also examined (HYP + Z1, Z1 + Z2, Z2 + Z3). Furthermore, during the procedures, the sheep were divided into three groups according to the body weight: Group (G) 1 (lighter), G2 (medium), and G3 (heavier). Finally, diagnostic outcomes were compared with necropsy findings. cUS obtained the highest values of sensitivity (Se) (91%), Specificity (Sp) (80%), and positive-zones (124/138, 90%), as compared to the other scans. cUS was also characterized by high values of Se and Sp and was able to identify a great number of positive-zones, when sheep were divided by body-weight groups. Similar performances were obtained in G1 by HYP (Se: 91%–Sp: 82%; 18/20, 90% of positive-zones) and HYP + Z1 scans (Se: 91%–Sp: 82; 90% of positive-zones, 18/20). Thus, in lighter breeds, the examination of HYP and HYP + Z1 scan windows could be considered reliable techniques for identifying the infected animals, while in heavier sheep the cUS still represents the best option.

In-House Developed ELISA Indicates High Prevalence of Anti-Echinococcus granulosus IgG in Sheep Population-An Update from Pakistan.

Cystic echinococcosis (CE) is a World Health Organization (WHO)-listed neglected tropical farm economy jeopardizing and public health concern disease. This study was aimed at furnishing sero-epidemiological baseline data of CE in sheep in Pakistan, where data are non-existent. For this purpose, two sheep-rich provinces of Pakistan were selected, and 728 sheep sera were collected using probability proportional to size (PPS) statistical technique. Epidemiological information was recorded on a questionnaire for the estimation of potential risk factors. The serum samples were analyzed for IgG antibodies against Echinococcus granulosus using an in-house-developed EgAgB-based ELISA kit. The overall seroprevalence recorded was 21.98% (160/728) in the tested sheep, suggesting higher seropositivity in sheep from Punjab (23.73%) as compared to Khyber Pakhtunkhwa (KPK) (19.04%). The overall apparent prevalence observed by this ELISA method was almost similar to the calculated true prevalence (21.77%). Prevalence was significantly different (p < 0.05) among sheep from different districts. Higher prevalence was found in females (22.54%, OR 1.41), age group > 5 years (29.66%, OR 1.64), crossbreeds (42.85%, OR 2.70), and sheep with pasture access (25.96%, OR 3.06). Being in age group > 5 years and having pasture access were the factors significantly associated with seropositivity (p < 0.05). This study provides serological evidence of E. granulosus infection in sheep and can be used as a model for ante-mortem screening of the sheep globally.

Bayesian Analysis of Three Methods for Diagnosis of Cystic Echinococcosis in Sheep.

Diagnosis of cystic echinococcosis (CE) in sheep is essentially based on necropsy findings. Clinical symptoms can be easily overlooked, while the use of immunological tests is still not recommended for an intra vitam diagnosis. This study assessed the performances of three post-mortem laboratory methods in the diagnosis of ovine CE. In the absence of a single and accurate test as a gold standard, the results of multiple analytical tests can be combined to estimate diagnostic performance based on a Bayesian statistical approach. For this purpose, livers (n = 77), and lungs (n = 79) were sampled from adult sheep and examined using gross pathology, histopathology and molecular analyses. Data from the three diagnostic methods were analyzed using a Bayesian latent class analysis model to evaluate their diagnostic accuracy in terms of sensitivity (Se), specificity (Sp), positive predictive value (PPV) and negative predictive value (NPV). The gross pathology examination revealed excellent diagnostic capabilities in diagnosing ovine CE with an Se of 99.7 (96.7–99.8), Sp of 97.5 (90.3–99.8), PPV of 97.6 (90.5–100), and NPV of 99.7 (96.5–100). The experimental design used in this work could be implemented as a validation protocol in a quality assurance system.

Cystic echinococcosis in the endemic island of Sardinia (Italy): has something changed?

Cystic echinococcosis (CE) is a disease of livestock and humans caused by the larval stage of the Echinococcus granulosus sensu lato (s.l.) species complex and occurs especially in areas where sheep breeding is widespread. This survey evaluates, after more than a decade since the last epidemiological survey, the current situation and trend of CE in sheep of the highly endemic island of Sardinia (Italy). From 2012 to 2018, out of a total of 1416 sheep, 65.3% were found positive for CE. Cyst fertility has increased slightly from 10.3% in 2006 (Scala et al. 2006) to 11.7% in the present survey (χ2 = 1.07; P = 0.30018) with a peak of 14.0% in the province of Sassari. Overall, 6815 hydatids were found and classified as sterile (26.2%), fertile (14.4%), caseous (5.3%), or calcified (54.1%) based on their morphology. All isolates were identified as E. granulosus sensu stricto (former G1/G3, strains). Results shown as the implementation of new European Council Regulations and in particular of 21/2004 that established a system for the individual identification and tracking of sheep could be a step forward towards a better control of animal movement, preventing illegal slaughtering on the island as well as in other countries experiencing similar problems. Even if at first sight the general situation of Sardinia regarding CE seems a little better compared with the past, our findings indicate that in some Provinces the epidemiological situation is not improved since 2006.

Molecular characterization of triosephosphate isomerase from Echinococcus granulosus.

Cystic echinococcosis (CE) is a zoonosis that can be caused by the larvae of Echinococcus granulosus; this disease occurs worldwide and is highly endemic in China. E. granulosus can produce energy by glycolysis as well as both aerobic and anaerobic respirations. Triosephosphate isomerase is a glycolytic enzyme present in a wide range of organisms and plays an important role in glycolysis. However, there has been little research on triosephosphate isomerase from E. granulosus (Eg-TIM). Here, we present a bioinformatic characterization and the experimentally determined tissue distribution characteristics of Eg-TIM. We also explored its potential value for diagnosing CE in sheep using indirect enzyme-linked immunosorbent assay (ELISA). Native Eg-TIM was located in the neck and hooks of protoscoleces (PSCs), as well as the tegument and parenchyma tissue of adult worms. The entire germinal layer was also Eg-TIM positive. Western blots showed that recombinant Eg-TIM (rEg-TIM) reacts with positive serum from sheep and had good immunogenicity. Indirect ELISA exhibited low specificity (53.6%) and low sensitivity (87.5%) and cross-reacted with both Taenia multiceps and Taenia hydatigena. Our results suggest that TIM may take part in the growth and development of E. granulosus. Furthermore, we determined that rEg-TIM is not a suitable serodiagnostic antigen for CE in sheep.

Expression, Tissue Localization and Serodiagnostic Potential of Echinococcus granulosus Leucine Aminopeptidase.

Echinococcus granulosus is the causative agent of cystic echinococcosis (CE), a widespread parasitic zoonosis. Leucine aminopeptidases (LAPs) of the M17 peptidase family have important functions in regulating the balance of catabolism and anabolism, cell maintenance, growth and defense. In this study, we presented a bioinformatic characterization and experimentally determined the tissue distribution characteristics of E. granulosus LAP (Eg-LAP), and explored its potential value for diagnosis of CE in sheep based on indirect ELISA. Through fluorescence immunohistochemistry, we found that Eg-LAP was present in the tegument and hooks of PSCs, the whole germinal layer and adult worm parenchymatous tissue. Western blotting results revealed that the recombinant protein could be identified using E. granulosus-infected sheep serum. The diagnostic value of this recombinant protein was assessed by indirect ELISA, and compared with indirect ELISA based on hydatid fluid antigen. The sensitivity and specificity rEgLAP-ELISA were 95.8% (23/24) and 79.09% (87/110), respectively, while using hydatid fluid as antigen showed the values 41.7% (10/24) and 65.45% (72/110). This is the first report concerning leucine aminopeptidase from E. granulosus, and the results showed that Eg-LAP belong to M17 peptidase families, and that it is involved in important biological function of E. granulosus. Furthermore, rEg-LAP is appropriate for diagnosing and monitoring CE in sheep in field. Development of a rapid test using rEg-LAP to diagnose sheep CE deserves further study.

Pathological and histochemical studies of the effects of cystic echinococcosis in sheep

A total of 2100 sheep, slaughtered or spontaneously dead, from various areas of Kashmir valley were screened for the presence of hydatidosis. Out of 2100 cases, 85 were positive for hydatidosis. Histopathologicallly, the cyst wall consisted of the inner germinal, middle lamellated, and outer fibrous layer. Inflammatory reaction around the cyst was variable and was characterized by an inner zone of loosely arranged fibroblasts infiltrated with mononuclear cells and densely packed fibroblasts mixed with mononuclear cells; and an outer layer of concentrically packed fibrous connective tissue. Mast cell reaction was observed mainly in the bronchiolar, peribronchiolar, and inflammatory zones in the lungs; while in the liver, mast cells were noted in portal triads and bile ducts and were least evident in the vicinity of the cysts. Qualitative increase was observed for acid mucopolysaccharides in fibroblasts, inflammatory cells, and bronchial epithelium, especially hyperplastic epithelial cells. Masson’s trichrome revealed intense formation of collagen fibers in the pericystic connective tissue. The calcareous corpuscles of the protoscolices were distinct when stained with combined Alcian blue PAS and toluidine stains but were not visible when stained with H & E and Masson’s trichrome stains.