May 27, 2022

Universal probe-based lateral flow immunoassay for the rapid quantitative detection of fumonisin B1

Fumonisin B1 (FB1) is a serious threat to the health of humans and animals. Herein, a lateral flow immunoassay based on universal detection probes that could combine with any mouse monoclonal antibody was applied to detect FB1 in corn and feed. Compared with that based on direct monoclonal antibody labeling, this assay maintained bioactivity and saved consumption of monoclonal antibodies with the indirect signal amplification effect.
The results indicated that this assay had higher sensitivity with a limit of detection (LOD) of 0.025 and 0.097 ng mL-1 (0.50 and 1.94 ng g-1 based on sample weight) in corn and feed, respectively. The detection range was about 1-50 ng mL-1 (20-1000 ng g-1 based on sample weight). In addition, the evaluation proved that it had good specificity, joplink Phospho-EGFR (Tyr1197) Antibody accuracy, precision, and applicability, and thus was suitable for the rapid and low-cost detection of fumonisin B1.

Seroepidemiology of leptospirosis in livestock and workers of high-risk occupation in Kurdistan

Leptospirosis is one of the major zoonotic infectious diseases which could cause disease in both animals and humans. Using ELISA is one of the serological tests that could be used in the detection of leptospirosis. Based on the different reports about the prevalence of leptospirosis in different parts of our country, we aimed to investigate the rate of Leptospira spp, among livestock and human in Kurdistan province.
ELISA assay was performed by ELISA kit (Novatec, Germany) for quantitative detection of anti-Leptospira IgG in human and IgM antibody and total antibodies (IgM and IgG) in serum samples of livestock.
In the present study, the seroprevalence in sheep, goats, and cows was (2/30) 6.7% [95% CI 0.8%,22.1%], (1/31) 3.2% [ 95% CI 0.08%,16.7%] and 0%, respectively. Also, the rate of anti-Leptospira was (1/51) 1.9% [95% CI 0.05%,10.4%] among 51 human participants.
the current study, provided some valuable information on the rate of leptospirosis in animal and human participants from west of Iran, which can be useful in terms of monitoring the disease in the area and helping the health care system to control the roots of bacterial transmission.

Cryptosporidium and Giardia in Livestock in Tigray, Northern Ethiopia and Associated Risk Factors for Infection: A Cross-Sectional Study

The occurrence and species/genotypes of Cryptosporidium and Giardia duodenalis infecting young livestock in selected districts of Tigray, Ethiopia were investigated, along with risks associated with infection. A total of 757 fecal samples were collected from calves, lambs, and goat kids from four rural districts in Tigray, and also from calves in periurban Mekelle, Tigray’s main city, and analyzed for Cryptosporidium oocysts and Giardia cysts. Farmers answered questionnaires regarding potential risk factors at sample collection.
Immunofluorescent antibody staining was used for parasite detection, and PCR at selected genes and sequencing of positive samples was used for molecular characterization. The occurrence of Cryptosporidium infection was 10, 9, and 4% in calves, lambs, and goat kids, respectively; equivalent figures for Giardia infection were 39, 32, and 21%. Molecular characterization of Cryptosporidium isolates revealed C. ubiquitum, subtype XIIa in all three host species; C. ryanae in calves and goat kids; C. andersoni and C. bovis were identified only in calves, and C. xiaoi was identified in lambs.
For Giardia, Assemblage E predominated in all host species, but among calf isolates, we also identified a few potentially zoonotic genotypes (assemblages A (AI) and Assemblage B). Periparturient care was shown to be a particularly relevant risk factor for infection, and infections were less likely to occur under extensive management systems. Our major findings were the widespread occurrence of both parasites in livestock, and the apparent lack of the most common zoonotic species.
Our results are discussed in relation to other relevant studies. As our study was conducted in Tigray, further investigation in different settings in Ethiopia could provide relevant information on transmission and zoonotic potential. In addition, given the dependency on healthy animals for the livelihoods of the population of Tigray, investigation of the effect of these common parasites on livestock productivity is important.

Evaluation of New Immunohistochemical Approaches for the Study of Kidney Tumors in Geriatric

Kidney malignancies are among the most deadly genitourinary tumors. It is more common in males and is often seen in people aged 60-70 years old. The incidence rate of kidney cancer seems to be increasing. One reason for this may be the fact that imaging techniques, such as computed tomography scans are more commonly used. These tests may lead to the accidental detection of more kidney cancers. Fortunately, kidney cancer is often detected in the early stages, when the tumor is small and confined to the kidney.
The objective of this study was the development of new diagnostic immunohistochemical methods.
Clinical examination material of 134 people, including 94 (70%) males and 40 (30%) females, were used in this study. Immunohistochemical staining of tryptase was carried out in compliance with the requirements using Anti-Mast Cell Tryptase antibodiesGoat anti-mouse antibodies #AS-M1-HRP were used as secondary antibodies, visualized with ImmPACTTM DAB Peroxidase Substrate Kit (#SK-4105) according to the instructions of the manufacturer.
The nuclei were counterstained with Mayer’s hematoxylin, and the sections were embedded in a permanent mounting medium. The immunohistochemical study showed an increase in both tryptase- and chymase-positive mast cells in the renal parenchyma, compared with the control group.
The number of mast cells with tryptase expression directly in the tumor was significantly less than the peritumoral localization.
A similar pattern was observed for chymase-positive mast cells as the content of the tumor was more than 10 times higher than the intratumoral arrangement. The histological and immunological characteristics did not differ in different age groups. The immunohistochemical method of research in the diagnosis of renal tumors plays an important diagnostic and prognostic value. It can assist pathologists in difficult and ambiguous cases to correctly diagnose renal tumors. This will make it possible to prescribe the correct treatment and predict the course of malignant tumor growth in patients.

Development of Immunochromatographic Assay for the Rapid Detection of Mycoplasma capricolum subsp. capripneumoniae Antibodies

Mycoplasma capricolum subsp. capripneumoniae (Mccp) is the cause of contagious caprine pleuropneumonia (CCPP), which is a highly significant respiratory disease in goats leading to significant economic losses in Africa and Asia. Currently available procedures for the diagnosis of CCPP have some limitations in sensitivity, specificity, operation time, requirement of sophisticated equipment or skilled personnel, and cost.
In this study, we developed a rapid, sensitive, and specific colloidal gold-based immunochromatographic assay (GICA) strip for the efficient on-site detection of antibodies against Mccp in the serum within 10 min. For the preparation of this colloidal GICA strip, recombinant P20 protein, the membrane protein of Mccp, was expressed by Escherichia coli prokaryotic expression system after purification was used as the binding antigen in the test.
The rabbit anti-goat immunoglobulin G labeled with the colloidal gold was used as the detection probe, whereas the goat anti-rabbit immunoglobulin G was coated on the nitrocellulose membrane as the control line. The concentration of the coating antibody was optimized, and the effectiveness of this colloidal GICA strip was evaluated.
Our results proved that the detection limit of the test strip was up to 1:64 dilutions for the Mccp antibody-positive serum samples with no cross-reactivity with other pathogens commonly infecting small ruminants,including goat pox virus, peste des petits ruminants virus, foot-and-mouth disease virus type A, or other mycoplasmas. Moreover, the colloidal GICA strip was more sensitive and specific than the indirect hemagglutination assay for the detection of Mccp antibodies. The 106 clinical serum samples were detected by the colloidal GICA strip compared with the complement fixation test, demonstrating an 87.74% concordance with the complement fixation test. This novel colloidal GICA strip would be an effective tool for the cost-effective and rapid diagnosis of CCPP in the field.

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