Antimicrobial activity of Lactobacillus strains of chicken origin against bacterial pathogens

This study was conducted to identify and evaluate the antimicrobial activity of some Lactobacillus isolates of chicken origin. Among 90 isolates 14 Lactobacillus species were distinguished using MALDI-TOF mass spectrometry and 16S-ARDRA. The dominant species was L. salivarius (34.4%), followed by L. johnsonii (23.3%), L. crispatus (13.3%) and L. reuteri (11.1%). All lactobacilli were screened for antimicrobial activity against wild-type strains of Salmonella enterica, Escherichia coli, and Clostridium perfringens. Results from the agar slab method showed that all Lactobacillus isolates were able to produce active compounds on solid media with antagonistic properties against these pathogens. The highest sensitivity to lactobacilli was observed in C. perfringens strains, and the lowest in E. coli. Lactobacillus salivarius exhibited particularly strong antagonism towards all of the indicator bacteria. Strains of L. ingluviei and L. johnsonii and one strain of L. salivarius (10d) selectively inhibited the growth of C. perfringens. No antimicrobial activity of many Lactobacillus isolates was observed when cell-free culture supernatant was used in a well diffusion assay. All Lactobacillus isolates exhibited the ability to produce H2O2 and proved to be hydrophobic (excluding one of L. salivarius). [Int Microbiol 19(1):57-67 (2016)]Keywords: Lactobacillus spp. · avian lactobacilli · antimicrobial activity · gut health · poultry pathogen

Učinak odabranih makrolida na neka obilježja goveđih leukocita

The aim of the study was to evaluate the effect of tylosin, tilmicosin and roxithromycin on viability, nitro blue tetrazolium reduction (NBT) assay, chemotaxis, apoptosis and oxidative stress in bovine leukocytes in vitro conditions. The material for the study consisted of blood collected into EDTA tubes from the external jugular vein of Holstein-Friesian cattle aged 1 week to 2.5 years, during routine veterinary examinations. In leukocytes the percent of viability, nitrate ion concentration (NO), metabolic activity (NBT, nitrotetrazolium blue reduction assay), chemotactic activity and apoptosis were determined. The results indicated a slight negative effect of these macrolides on the viability of the leukocytes, and confirmed the ability of macrolides to induce apoptosis in leukocytes in vitro. These results indicate that all of the macrolides investigated exhibit a modulatory effect on the functions of leukocytes isolated from cattle of different ages. The strongest inhibitory effect on the NBT reduction assay and chemotaxis of the leukocytes was exhibited by roxithromycin, which at the same time had the least negative effect on the leukocytes.Cilj je ovog istraživanja bio procijeniti učinak tilozina, tilmikozina i roksitromicina na preživljavanje, test redukcije “nitro-tetrazol-modrog” (NBT), kemotaksiju, apoptozu i oksidacijski stres goveđih leukocita u uvjetima in vitro. Prilikom rutinskog veterinarskog pregleda, u epruvete s EDTA bila je uzeta krv iz jugularne vene goveda holštajnsko-frizijske pasmine u dobi od tjedan dana do 2,5 godine. Određen je postotak preživljavanja leukocita, koncentracija iona nitrata (NO), metabolička aktivnost (test redukcije “nitro-tetrazol-modrog”, NBT), kemotaksijska aktivnost i apoptoza. Rezultati su pokazali blagi negativni učinak spomenutih makrolida na preživljavanje leukocita i potvrdili sposobnost makrolida da potaknu njihovu apoptozu in vitro te naznačuju da svi pretraženi makrolidi imaju modulacijski učinak na funkciju leukocita goveda različite dobi. Najjači inhibicijski učinak na test redukcije NBT i kemotaksiju leukocita pokazao je roksitromicin, koji je istodobno imao najmanje negativan učinak na leukocite

AN IMMUNO-ELECTROPHORETICAL ANALYSIS OF SOLUBLE BLOOD PROTEINS OF COILED AND UPRIGHT LARVAE OF THE WORKER HONEY BEE, APIS MELLIFERA

International audienc

Bacteriophage therapy to combat bacterial infections in poultry

Abstract Infections in poultry are an economic and health problem in Europe and worldwide. The most common infections are associated with salmonellosis, colibacillosis, campylobacteriosis, and others. The prevalence of Campylobacter-positive poultry flocks in European countries varies from 18% to 90%. In the United States, the prevalence of infected flocks is nearly 90%. A similar percentage of infection has been noted for salmonellosis (about 75–90%) and E. coli (90–95%). The occurence of Clostridium perfringens is a major problem for the poultry industry, with some estimates suggesting colonization of as many as 95% of chickens, resulting in clinical or subclinical infections. In the US, annual economic losses due to Salmonella infections run from $1.188 billion to over$11.588 billion, based on an estimated 1.92 million cases. Similar costs are observed in the case of other types of infections. In 2005 economic losses in the the poultry industry due to mortalities reached 1,000,000 USD. Infections caused by these pathogens, often through poultry products, are also a serious public health issue. The progressive increase in the number of multi-drug resistant bacteria and the complete ban on the use of antibiotics in livestock feed in the EU, as well as the partial ban in the US, have led to the growth of research on the use of bacteriophages to combat bacterial infections in humans and animals. The high success rate and safety of phage therapy in comparison with antibiotics are partly due to their specificity for selected bacteria and the ability to infect only one species, serotype or strain. This mechanism does not cause the destruction of commensal bacterial flora. Phages are currently being used with success in humans and animals in targeted therapies for slow-healing infections. They have also found application in the US in eliminating pathogens from the surface of foods of animal and plant origin. At a time of growing antibiotic resistance in bacteria and the resulting restrictions on the use of antibiotics, bacteriophages can provide an alternative means of eliminating pathogens

Molecular and Serological Characteristics of Avian Pathogenic Escherichia coli Isolated from Various Clinical Cases of Poultry Colibacillosis in Poland

Escherichia coli infections are a major problem in modern poultry production. Avian pathogenic E. coli (APEC) strains have several mechanisms that enable them to colonize various ecosystems. In this study, 290 E. coli isolates were recovered from clinical cases of colibacillosis in chicken and turkey broilers and from laying and breeding hens. The samples were taken from organs with pathological changes suggesting colibacillosis. The lesions were assigned to three groups depending on their advancement, of which the largest (60% of the isolates) was group 3, with the most extensive changes. The most common serotype was shown to be O78 (14%). The most frequently detected gene among those tested was iss, while papC was the least prevalent. An analysis of the number of genes present per isolate revealed that the presence of four genes was the most common (22%), while only 1% of the strains tested had all eight genes. The most frequently detected genes for each serotype were iss and iucD for O78; irp2 and cvi/cva for O1; irp2, iucD, and iss for O2, and iss and iucD for O8, for which the least frequent was papC. All O18 serotype strains had the iss gene, while none had the vat gene

Isolation and Characterization of Lytic Properties of Bacteriophages Specific for M. haemolytica Strains.

The objective of this study was isolation and morphological characterization of temperate bacteriophages obtained from M. haemolytica strains and evaluation of their lytic properties in vitro against M. haemolytica isolated from the respiratory tract of calves.The material for the study consisted of the reference strain M. haemolytica serotype 1 (ATCC®) BAA-410™, reference serotypes A1, A2, A5, A6, A7, A9 and A11, and wild-type isolates of M. haemolytica. Bacteriophages were induced from an overnight bacterial starter culture of all examined M. haemolytica strains treated with mitomycin C. The lytic properties and host ranges were determined by plaque assays. The morphology of the bacteriophages was examined in negative-stained smears with 5% uranyl acetate solution using a transmission electron microscope. The genetic analysis of the bacteriophages was followed by restriction analysis of bacteriophage DNA. This was followed by analysis of genetic material by polymerase chain reaction (PCR).Eight bacteriophages were obtained, like typical of the families Myoviridae, Siphoviridae and Podoviridae. Most of the bacteriophages exhibited lytic properties against the M. haemolytica strains. Restriction analysis revealed similarities to the P2-like phage obtained from the strain M. haemolytica BAA-410. The most similar profiles were observed in the case of bacteriophages φA1 and φA5. All of the bacteriophages obtained were characterized by the presence of additional fragments in the restriction profiles with respect to the P2-like reference phage. In the analysis of PCR products for the P2-like reference phage phi-MhaA1-PHL101 (DQ426904) and the phages of the M. haemolytica serotypes, a 734-bp phage PCR product was obtained. The primers were programmed in Primer-Blast software using the structure of the sequence DQ426904 of reference phage PHL101.The results obtained indicate the need for further research aimed at isolating and characterizing bacteriophages, including sequence analysis of selected fragments. Moreover, standardization of methods for obtaining them in order to eliminate M. haemolytica bacteria involved in the etiopathogenesis of BRDC is essential