ABSTRACT
Infection with Gasterophilus intestinalis (botfly) larvae often occurs in horses. The aim of the study was to isolate the larvae of G. intestinalis and evaluate the serum and salivary humoral immune response using self-developed ELISA in G. intestinalis infected horses. Blood serum or saliva samples were taken from 125 infected horses and 54 uninfected slaughtered horses. The antigens from G. intestinalis larvae were used for development of ELISA in order to evaluate the intensity of G. intestinalis IgG, IgM, and IgA antibody reactivity in the serum or saliva of naturally infected horses and horses without larvae in the gastrointestinal tract (control group). Serum antibodies against second and third larvae's stadium antigens reacted significantly more intensively in infected than in healthy horses in IgG (p ≤ 0.001; p ≤ 0.05, respectively) and IgA (p ≤ 0.05;p ≤ 0.001, respectively) classes. Salivary IgG and IgA specific's antibody reactivity was significantly higher in horses with moderate (p ≤ 0.01) and severe infection (p ≤ 0.001) compared to the healthy horses. The determination of the G. intestinalis IgG and IgA antibody activity in saliva and serum may be used for detecting horses moderately and severely infested with larvae.
Subject(s)
Diptera , Horse Diseases , Animals , Horses , Larva , Immunity, Humoral , Serum , Seasons , Diptera/physiology , Immunoglobulin A , Immunoglobulin G , Immunoglobulin MABSTRACT
Fungi belonging to the Cryptococcus neoformans/C. gattii species complex (CNGSC) are pathogens causing severe infections in humans and animals, that for humans may result in a mortality rate ranging up to 70%. The CNGSC is divided into eight major molecular types, that may differ in their virulence and susceptibility. In order to fully understand the epidemiology of cryptococcosis, it is important to study the world distribution and population structure of these pathogens. The present study is the first presenting a population of strains isolated in Poland and one of the few using a multi-species animal group as a source of the specimen. The pathogen was present in 2.375% of the tested animals. The URA5-RFLP and MALDI-TOF MS analyses have revealed that the population consisted exclusively of C. neoformans strains, with a predominance of major molecular type VNIV (C. neoformans var. neoformans). The MALDI-TOF MS was used to perform the CNGSC strains identification on both the species and sub-species level. Despite the fact that the animals providing the specimens were not treated with 5-fluorocytosine, around 10% of the tested population presented MIC values exceeding 64 mg/L, indicating the existence of the 5-fluorocytosine-resistant strains in the environment.
Subject(s)
Animal Diseases/epidemiology , Animal Diseases/microbiology , Cryptococcosis/veterinary , Cryptococcus neoformans/classification , Animal Diseases/history , Animals , Antifungal Agents/pharmacology , Cryptococcus neoformans/drug effects , Cryptococcus neoformans/genetics , Cryptococcus neoformans/isolation & purification , History, 21st Century , Microbial Sensitivity Tests , Molecular Typing , Mycological Typing Techniques , Poland/epidemiology , Polymorphism, Restriction Fragment Length , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
Heat Shock Proteins (HSP) are highly conserved proteins that are widely spread throughout all organisms. They function in the cytoplasm as chaperones; however, they could be expressed on the cell surface. It has been shown that Hsp60 obtained from gram-negative bacteria are able to stimulate cells of the acquired and innate immune system. The aim of this study was the evaluation of the immunogenic properties of recombinant Hsp60 proteins derived from four common pathogenic bacteria: Escherichia coli, Histophilus somni, Pasteurella multocida and Salmonella Enteritidis. The analysis of the humoral immune response in DBA/2J mice hyperimmunized with selected rHsp60 revealed high levels of IgG rHsp60-antibody with the predominance of the IgG1 subclass, in the reaction with both homologous and heterologous antigens. The presence of IgG2a and IgG2b was also observed; however, no antibodies of subclass IgG3 were detected. The comparison of plasma IgG antibody reactivity of mice immunized with two different doses of rHsp60 (10/20⯵g) showed that the lower dose was sufficient to induce a strong humoral response. The reactivity of the IgG rHsp60-antibody with whole bacterial cells showed a significantly higher reaction with H. somni compared with other pathogens. It was demonstrated that the addition of all rHsp60 with polymyxin B to the culture medium stimulated splenocytes isolated from hyperimmunized mice to release IL-1ß and IL-6. As a strong stimulator of the immune system, bacterial-origin Hsp60 seems to be an interesting potential component of subunit vaccines aimed at the development of protection for animals during infections caused by gram-negative bacteria.
Subject(s)
Antibodies, Bacterial/blood , Antibody Formation , Antigens, Bacterial/immunology , Chaperonin 60/immunology , Immunization , Animals , Antigens, Bacterial/genetics , Chaperonin 60/genetics , Cytokines , Female , Gene Expression Regulation, Bacterial , Gram-Negative Bacteria/immunology , Immunity, Humoral , Immunity, Innate , Immunoglobulin G/blood , Interleukin-1beta/metabolism , Interleukin-6/metabolism , Mice , Mice, Inbred DBA , Polymyxin B/pharmacology , Recombinant Proteins/immunology , Recombinant Proteins/isolation & purification , Tumor Necrosis Factor-alphaABSTRACT
The analysis of the data shows that the growth of roe deer from the eastern part of Greater Poland (Wielkopolska) is very similar to that of specimens from other areas in Poland. A comparison with excavated bone material revealed only slight and fluctuating differences. The roe deer from the areas to the west of Poland are larger and those from the areas to the east of the country are the largest.
