Detection of Anti-Echinococcus granulosus Antibodies in Humans: An Update from Pakistan

Human cystic echinococcosis (CE) is a zoonotic disease caused by the larval stage of Echinococcus granulosus sensu lato that causes economic losses by affecting livestock and also poses a public health threat worldwide. The present study is the first retrospective report on the seroprevalence of anti-E. granulosus antibodies in humans in Pakistan. The study used data from 93 blood analysis reports of patients suspected of having CE from different medical centers in Lahore, Pakistan. Out of 93 sera samples, 20 (21.5%) were seropositive, and higher seropositivity (17.2%) was recorded with the indirect hemagglutination test (IHA) than with enzyme-linked immunosorbent assay (ELISA). The findings indicated that age, gender, and year had no significant relationship with the seropositivity of CE. The current study provides directions towards the management of the disease in the near future in Pakistan.

Isolated Renal Hydatid Cyst in a 6-Year-Old Boy: A Case Report

Hydatid cyst (HC) is a parasitic infection transmitted by oral ingestion of Echinococcus granulosus eggs. Isolated kidney involvement is extremely rare. It is even less common in children. We present a case of isolated renal HC in a 6-year-old boy from Şanlıurfa, southeast Turkey who complained of abdominal pain and distension. Cystectomy was performed with a flank incision using the extraperitoneal approach. No recurrence was detected in the postoperative 6-month follow-up. There are not enough data about the efficacy of medical treatment in renal HC. Therefore, medical treatment should be considered pre- and post-operatively to prevent dissemination, rather than being used as a primary treatment. Kidney-sparing surgery should be the first choice in patients with isolated renal HC. However, nephrectomy is recommended for nonfunctioning kidneys, large cysts thought to be connected with the collecting system, and cysts with suspicious tumor. In societies where HC disease is endemic, renal HC should be considered in children with cysts located in kidney, even if the indirect hemagglutination test is negative.

Presence of atypical genotypes of Toxoplasma gondii isolated from cats in the state of Bahia, Northeast of Brazil

In this study, 20 blood, heart, and brain samples were collected from euthanized cats at the Zoonosis Control Centers and Veterinary Clinics in the state of Bahia, Brazil. The sera were examined for anti-T. gondii antibodies using the indirect hemagglutination test. The brains and hearts of seven seropositive cats were ground, and peptide digestion was performed for bioassay in mice. Toxoplasma gondii was isolated in 5/7 (71.42%) of seropositive cats. In these isolates, the parasite was genotyped using the Polymerase chain reaction, associated with the DNA fragment polymorphism obtained by restriction enzyme PCR-RFLP technique with 11 markers (SAG1, 5’-SAG2, 3’-SAG2, alt. SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico, and CS3) and 15 microsatellite markers (TUB-2, W35, TgM-A, B18, B17, M33, IV.1, XI.1, M48, M102, N60, N82, AA, N61, N83). The analysis of the isolates by PCR-RFLP revealed five distinct genotypes. Three of these genotypes have never been reported before; one corresponded to the TgDgCo13 genotype, and one incomplete genotype. In genotyping analysis using microsatellite markers, it was observed that the isolates showed atypical alleles in the typing and fingerprint markers. This revealed five atypical genotypes. The typing marker B17 showed the highest degree of atypia. This study is the first to report the genotyping of T. gondii obtained from naturally infected cats in Bahia, Northeast Brazil. The genotypes found in this study were different from those found in other studies conducted in Bahia, which included different species of animals. None of the clonal lineages I, II, or III were found. This study demonstrates the diversity of T. gondii in the study region, with the presence of unusual genotypes, reaffirming the genetic variability of the parasite in Brazil.

Development of new antigens for serologic diagnosis of animal trypanosomosis and their use for epizootic monitoring

The purpose of the research is developing a method for obtaining erythrocyte antigens containing and not containing Trypanosoma equiperdum and T. evansi DNA, which can later be used in serological reactions to differentiate these types of Trypanosoma.Materials and methods. The studies were conducted in the Protozoology Laboratory and the Vyshnevolotsk Branch of the Federal State Budget Scientific Institution “Federal Scientific Centre VIEV RAS”, as well as livestock farms of the Russian Federation and other countries using clinical, microscopic, hematological, parasitological, biomolecular and serological methods.Results and discussion. Studies carried out for the first time have shown that it is possible to use erythrocyte antigens containing the T. equiperdum and T. evansi DNA obtained after 3-fold administration to mice and rabbits of a mixture of trypanosomal antigen with addition of 1.0 ml of an adjuvant (aluminum hydroxide), and bleeding of animals at 25 to 30 days. The formed precipitate was used as an antigen for serological tests. Experiments have shown that blood for preparation of positive serum can be taken when antibodies are in titers of 1:20 in the Prolonged Complement Fixation Test, and at least 1:400 in the Indirect Hemagglutination Test and ELISA, and for negative serum when horse blood serum reacts negatively with antigens of T. equiperdum and T. evansi in the Prolonged Complement Fixation Test, Indirect Hemagglutination Test and ELISA. The test systems of the Prolonged Complement Fixation Test, Indirect Hemagglutination Test and ELISA prepared by us with antigens containing and not containing T. equiperdum and T. evansi DNA resulted in creating a universal test system (Indirect Hemagglutination Test) for differentiating T. equiperdum from T. evansi.

Role of a schistosoma haematobium specific microRNA as a predictive and prognostic tool for bilharzial bladder cancer in Egypt

Urinary bladder cancer is a common malignancy in Egypt, thus reliable methodologies are required for screening and early detection. In this study, we analyzed the gene expression of a Schistosoma hematobium specific microRNA “Sha-miR-71a” and mitogen-associated protein kinase-3 (MAPK-3) in the urine samples of 50 bladder cancer patients and 50 patients with benign bilharzial cystitis. Fifty control subjects were also tested. Indirect hemagglutination test (IHA) diagnosed 70% of studied cancer cases as bilharzial associated bladder cancer (BBC), while histopathological examination detected only 18%. Urinary Sha-miR-71a & MAPK-3 revealed enhanced expression in BBC (p-value = 0.001) compared to non-bilharzial bladder cancer (NBBC) cases. Patients with chronic bilharzial cystitis exhibited a significant increase in gene expression compared to those with acute infection (p-value = 0.001). Sha-miR-71a and MAPK-3 showed good sensitivity and specificity in the diagnosis of BBC when analyzed by the receiver operating characteristic (ROC) curve. They were also prognostic regarding malignancy grade. Both biomarkers showed a positive correlation. Our results revealed that IHA is a reliable test in the diagnosis of bilharziasis associated with bladder cancer, and that Sha-miR-71a and MAPK-3 provide non-invasive specific biomarkers to diagnose BBC, as well as a potential role in testing bilharzial patients for risk to develop cancer.

Indirect Hemagglutination Test for Detection of Antibodies to Toxoplasma gondii in Venezuelan felids

Current knowledge of Toxoplasma gondii infection in Venezuelan ecosystems is limited. T. gondii is a ubiquitous intracellular protozoan parasite. Mammals and birds are intermediate hosts and felid species are definitive hosts. In most human altered habitats, the domestic cat is the predominant definitive host. Cats are important in the epidemiology of T. gondii infection because they are the only hosts that can excrete the environmentally resistant oocysts. Other carnivores can be infected by the consumption of tissue cysts when feeding on infected animals and by incidental ingestion of oocysts from environmental contamination. This study aimed to quantify the values of antibodies for T. gondii in blood serum of some felids species by means of the technique of Indirect Hemoagglutination. In the present study, seropositivity of T. gondii was determined in serum of 35 animals (22 stray cats and 13 wild cats) from Venezuela, South America. Antibodies to T. gondii were assayed by the indirect hemagglutination test and found in 21 of 22 (95.45 %) stray catstiters of 1:64 in four, 1:128 in four, 1:256 in one, 1:512 in one, 1:1024 in three, and 1:2048 or higher in eight. In 4 of 6 (66.67 %) ocelots titers of 1:64 in one, 1:256 in one, 1:1024 in one, and one with titers 1:2048. In 3 of 4 (75.00 %) jaguars titers of 1:512 in one, and two with titers 1:2048. The Kruskal-Wallis test showed a statistically significant difference between species (H = 6.983, p = 0.03).

Analysis of the epizootic situation and improvement of the scheme for the specific prevention of anthrax

The article shows the epizootic situation for three periods (decades): from 1990 to 1999, from 2000 to 2009 and from 2010 to 2019. As a result of the studies, it has been established that since 1990, the 26 sporadic outbreaks of anthrax have occurred in the territory of the republic, 13 of them in territories where the disease was not previously recorded, which indicates the spread of the pathogen in the environment. Due to the special situation of anthrax, which consists of a large number of unidentified anthrax cattle burial grounds, as well as high density and activity of dysfunctional sites, the republican veterinary service recommended vaccinating an adult cattle population twice a year with an interval of 6 months: in spring and autumn, to create the maximum percentage of immune animals. In addition, a method has been introduced in the republic for assessing specific prophylaxis of anthrax, based on the detection of specific antibodies in the blood serum of an immunized livestock by an indirect hemagglutination test. This method allows the detection of animals that were skipped during vaccination and tolerant animals. Thanks to the joint work of veterinarians and scientists who improved the specific scheme for the prevention of anthrax, the republic managed to reduce the number of outbreaks of infection by 15 times, which allows us to predict further improvement in the epizootic situation for this infection.

Production testing of antigenicity and immunogenicity of bivalent inactivated vaccine salmonellosis vaccine

Scientists of both human and veterinary medicine combine their potential to develop new or improve old instruments in order to rein the problem of foodborne salmonellosis in Ukraine. According to the experience of European poultry industry, the most effective measure for the prevention of avian salmonellosis is total vaccination against salmonellosis of laying hens and breeding birds. In our country, the epizootic situation of salmonellosis of animals and poultry is consistently favorable. At the same moment nearly 90% of foods born Salmonella outbreaks are of poultry and egg products origin. In other words the source of major source of Salmonella agent is avian origin. Currently no vaccines of native origin have been registered in Ukraine. Although there have been numerous attempts to develop a vaccine against avian salmonellosis. The purpose of our work is to evaluate the antigenicity and immunogenicity of the two experimental series of bivalent inactivated emulsified vaccine against avian salmonellosis in production conditions. During the examination of the vaccine in the poultry farm it was found that for 21 days after the re-introduction of the vaccine, the titers of antibodies to the mono-antigens S. Typhimurium, Enteritidis and Gallinarum in the Agglutination Test (AT) and Indirect Hemagglutination Test (IH) were: 1 : 640–1280 to 1 : 2560–5120 respectively. This indicates high antigenicity of the vaccine. No significant difference between the levels of antibodies to Typhimurium and Enteritidis antigens was detected neither in AT nor IH. At the same time the levels of antibodies to mono-antigen Gallinarum were markedly lower in both reactions (1 : 160–1 : 320 – in AT and 1 : 320–1 : 1280 – in IH), but high enough to indicate that the vaccine creates a tense cross-humoral immunity to Salmonella surface antigens of Gallinarum serovar. The results of study of immunogenicity of the vaccine show that the vaccine is highly immunogenic. It means that after control infection of vaccinated hens none of tested Salmonella strains (S. Typhimurium and S. Enteritidis) were isolated from any organs, whereas in the control non-vaccinated group of birds both Salmonella test-strains were isolated from all organs. The obtained results provide a basis for further phases of the vaccine estimating followed by its registration in the prescribed manner.