{"id":5448,"date":"2022-10-13T20:13:51","date_gmt":"2022-10-13T18:13:51","guid":{"rendered":"https:\/\/veterinarska-stanica-journal.hr\/?post_type=article&p=5448"},"modified":"2022-10-13T20:13:51","modified_gmt":"2022-10-13T18:13:51","slug":"signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals","status":"publish","type":"article","link":"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals","title":{"rendered":"Signs of pathogenicity by Pasteurella multocida<\/em> in different species of animals"},"content":{"rendered":"

\"\"<\/p>\n

S. Boianovskiy<\/strong>, V. Ushkalov<\/strong>*, L. Vygovska<\/strong>, T. Mazur<\/strong>, L. Ishchenko<\/strong>, K. Rudnieva<\/strong>, A. Ushkalov<\/strong> and V. Melnyk<\/strong><\/p>\n

\n
Serhii BOIANOVSKIY<\/strong>, Valerii USHKALOV<\/strong>*, (Corresponding author, e-mail: ushkalov63@gmail.com), Lilia VYGOVSKA<\/strong>, Tatyana MAZUR, Liudmyla ISHCHENKO<\/strong>, National University of Life and Environmental Sciences of Ukraine 15 Heroyiv Oborony str., Kyiv, Ukraine; Kateryna RUDNIEVA<\/strong>, Kyiv Regional Clinical Hospital, 1 Baggovutivska str., Kyiv, Ukraine; Artem USHKALOV<\/strong>, Main administration of state service of Ukraine on food safety and consumer protection in Kharkiv reg. Ukraine; Volodymyr MELNYK<\/strong>, National University of Life and Environmental Sciences of Ukraine 15 Heroyiv Oborony str., Kyiv, Ukraine<\/div>\n
\"\"<\/a>https:\/\/doi.org\/10.46419\/vs.54.3.5<\/a><\/div>\n<\/p>\n

<\/a><\/p>\n

\n
Abstract<\/a>Introduction<\/a>Materials and methods<\/a>Results<\/a>Discussion<\/a>Conclusion<\/a>References<\/a>Sa\u017eetak<\/a><\/span><\/div>\n<\/div>\n

<\/a> ▲<\/a><\/p>\n

\n

Abstract<\/h2>\n
\n

A significant number of microorganisms in natural and artificial environments exist in a structured formation \u2013 biofilm. This formation attaches to a certain surface, particularly the epithelium. The ability to form a similar structure has been observed in Pasteurella multocida<\/em>, the causative agent of anthropozoonoses that affect domestic and wild animals, birds, companion animals and humans. The spectrum of pathogenetic action of P. multocida<\/em> is wide and associated with the development of respiratory and multisystemic pathology, bacteraemia and other manifestations. Timely detection of P. multocida<\/em> and treatment of the diseases it causes in farm and domestic animals is important to limit economic losses and improve social security. The main objective of this study was to determine the pathogenicity of P. multocida<\/em>, its ability to form a biofilm, its resistance to antibiotics, and to identify the genes responsible for the formation of dermonecrotic toxin and biofilm formation. The paper presents the results of a study of 11 isolates of P. multocida<\/em>: six isolates (54.5%) from rabbits, two isolates (18.2%) from dogs, two isolates (18.2%) from cats, and one isolate from pigs (9.2%). In all isolates, the gene ptfA<\/em> was detected. This gene encodes the formation of type 4 fimbriae and participates in the formation of the biofilm, and the studied cultures in vitro<\/em> formed a biofilm of different densities. The genome of eight isolates (72.7%) included the toxA<\/em> gene (provides the formation of dermonecrotic toxin), while 45.4% of isolates had a complete set of the studied signs of pathogenicity, both in phenotypic (biofilm formation, mortality for laboratory animals) and genotypic (presence of toxA<\/em>, ptfA<\/em>) traits, and three isolates (27.3%) showed signs of multidrug resistance. The virulence of the toxA<\/em>-negative isolates of P. multocida<\/em> was lower than in toxA<\/em>-positive isolates. The culture with the highest virulence (0.5 x 101<\/sup> CFU) and extreme resistance to antibiotics formed a biofilm of the highest density. The association of the gene in the biofilm-producing mechanism needs further evaluation, and further research is needed to identify the relationships between pathogens in Pasteurella multocida<\/em> isolates from different species of animals and humans.<\/p>\n

Key words:<\/strong> biofilm; Pasteurella multocida; antibiotic resistance; toxA; ptfA<\/em><\/p><\/blockquote>\n

<\/a> ▲<\/a><\/p>\n

Introduction<\/h2>\n
\n

The evolution of infectious diseases requires the study of the biological properties of pathogens: morphological, enzymatic, molecular genetics, pathogenic, susceptibility to antibacterial agents, and demands a search for new alternatives for their treatment and prevention. An important area of research is the development of measures and methods for their prevention, including zoonoses \u2013 diseases transmitted from animals to humans through direct contact or through food, water, and the environment (Bruchmann et al<\/em>., 2021; Guan et al<\/em>., 2021).<\/p>\n

A large variety of microorganisms, in addition to their capacity to attach to surfaces, also produce an extracellular polymeric substance. This substance forms a thin layer around cells known as biofilm, a structure that comprises an extracellular polymeric substance and the bacterial cells within it. This ability to form biofilm leads to the major pathogenic factor of bacterial infections.
\nBiofilm protects against attacks from the immune system and against antibiotic treatment, hindering the eradication of these infections (Donlan and Costerton, 2002; Coenye and Nelis, 2010; Petruzzi et al<\/em>., 2018).<\/p>\n

Bacteria in biofilm mode undergo conspicuous changes in their genetic and phenotypic expression by expressing many novel proteins, constituted by the outer membrane and heat shock proteins (Petruzzi et al<\/em>., 2018; Guan et al<\/em>., 2021). Biofilms could cause chronic and recrudescent infections, that are difficult to control by treatment (Welin, 2004).
\nThe increased resistance of biofilms is explained by several factors: 1) different rates of diffusion of substances; 2) the accumulation of extracellular enzymes in the matrix that have a destructive effect on antibiotics; 3) inaccessibility of bacteria due to adhesion; 4) the resistant properties of the cells themselves (Jamal et al<\/em>., 2018; Guan et al<\/em>., 2019; Petruzzi et al<\/em>., 2020).<\/p>\n

Various adhesins helps gram-negative bacteria in the colonisation of host tissues. One of these adhesins is type 4 fimbriae (pili). These structures allow gram-negative bacteria to colonise epithelial surfaces. This pili structure can be observed in Pasteurella multocida<\/em> strains A, B and D. The type 4 fimbrial subunit protein (\u0440tfA<\/em>) was identified as an 18-kDa protein isolated from whole membrane fractions (Doughty et al<\/em>., 2000).<\/p>\n

Biofilm formation of P. multocida<\/em> has become a new perspective of its virulence study, since it is a respiratory zoonotic pathogen and its ability to form biofilm could possibly be one of its virulence factors for survival inside the host (Steen et al<\/em>., 2010; Rajagopal et al<\/em>., 2013; Peng et al<\/em>., 2017; Guan et al<\/em>., 2020).<\/p>\n

The main objective of this study was to describe biofilm formation of P. multocida<\/em>, methods for its detection and the presence of genes responsible for dermonecrotic toxin and biofilm formation.<\/p>\n

<\/a> ▲<\/a><\/p>\n

Materials and methods<\/h2>\n
\n

This study was performed at the Ukrainian Laboratory of Quality and Safety of Agricultural Products and the Department of Epizoology, Microbiology and Virology of the National University of Life and Environmental Sciences of Ukraine (Kyiv, Ukraine) during 2019\u20132020. Research conducted with the use of animals in accordance with the requirements of the Ukraine Law On Protection of Animals from Cruelty, and Directive 2010\/63\/EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes.<\/p>\n

We used a total of 11 cultures of P. multocida<\/em>, isolated from various sick and clinically healthy animals.
\nCultures of P. multocida<\/em> were isolated as a result of bacteriological examination of pathological material (from heart and liver blood) from 24 dead rabbits, 5 piglets, and the pharyngeal and tonsil smears from 12 clinically healthy dogs and 9 cats (veterinary clinic patients). Bacteriological studies were performed in accordance with the current requirements for bacteriological diagnosis of pasteurellosis in animals and the Manual of Diagnostic Tests and Vaccines for Terrestrial Animals.<\/p>\n

LD100<\/sub> was determined by subcutaneous injection of test cultures in doses of 0.5×101<\/sup>\u20130.5×109<\/sup> CFU, the results were recorded for 72 hours. The LD100<\/sub> was taken as the minimum amount of the studied culture, which caused 100% death of experimental animals.<\/p>\n

A total of 11 animal P. multocida<\/em> isolates were examined for virulence-associated and biofilm-associated genes (ptfA<\/em> and toxA<\/em>) using various polymerase chain reaction (PCR) methods reported elsewhere (Curtis, 1985).<\/p>\n

Antibiotic susceptibility of P. multocida<\/em> cultures was determined using the disc-diffusion method using discs produced by HiMedia (India), and Mueller-Hinton II agar with 5% horse blood produced by GRASO (Poland). Studies and the interpretation of results were performed according to EUCAST recommendations (version 12, available at: https:\/\/www.eucast.org<\/a>), which provide specific recommendations for determining the susceptibility of P. multocida<\/em> (Magiorakos et al<\/em>., 2012).<\/p>\n

The ability to form biofilms in the derived isolates was determined and the results interpreted (Kukhtyn and Krushelnytska, 2014). This study was performed using sterile polystyrene Petri dishes (Greiner Bio-One GmbH, Germany) of d=100 mm, in which 10 mL brain-heart infusion broth (HiMedia, India) and 1 mL inoculum with a cell content of 0.5 were added to the MacFarland daily culture of studied P. multocida<\/em> isolates.
\nThe plates were cultured in a thermostat at a temperature of 37\u00b0C for 24 hours, the residues of the nutrient medium were carefully removed, the planktonic forms were washed three times with a sterile phosphate buffer solution (KH2<\/sub>PO4<\/sub>\u00b7Na2<\/sub>PO4<\/sub>\u00b7H2<\/sub>O), pH 7.2\u20137.4. The Petri dishes were air-dried and 10 mL 96% ethanol was added to fix the formed biofilms. The fixation exposure was 10 minutes. The fixing liquid was drained and then the Petri dishes were divided into two parts: the first was stained with 0.1% alcohol solution of crystal violet for 10 min, and the second was stained with a mixture of saturated aqueous Congo red for 15 min. Plates were washed three times with sterile phosphate buffer solution (pH 7.2) and dried. Contributed to 10,0 mL 96% ethanol and placed on a shaker for shaking for 30 min, were then pipetted transferred to a cuvette and determined the optical density spectrophotometricall on an Evolution 300 spectrophotometer (Thermo Fisher Scientific, USA) at a wavelength of 570 nm for Petri dishes with solution of crystal violet and 495 nm for Petri dishes with solution of Congo red. The density of the formed biofilm was determined by measuring the adsorption level of the dye with ethanol measured in units of optical density (OD) using a spectrophotometer.<\/p>\n

When the value of optical density is less than 0.1, strains were not considered to form a biofilm. At optical densities from 0.1 to 0.49 the ability to form a film was considered low, from 0.5 to 1.0 medium with medium density, and at values above 1.0 high with high density (Ewers et al<\/em>., 2000).<\/p>\n

Genomic DNA from P. multocida<\/em> cultures was isolated by the express method. For this, the lyophilized mass of the culture of P. multocida<\/em> was dissolved in 1 mL sterile buffered peptone water (HiMedia, India) and centrifuged at 13,500 rpm for 2 min and the supernatant was removed. The bacterial pellet was resuspended in 200 \u03bcL TE buffer and incubated in a thermostat at 95\u00b0C for 5 minutes. Cell debris was precipitated by centrifugation at 5000 rpm for 2 min and 180 \u03bcL supernatant was taken for the PCR.
\nThe DNA concentration was measured on a Biofotomer spectrophotometer (Eppendorf, Germany) at a wavelength of 260 nm. The amplification reaction was performed in a reaction mixture with a volume of 25 \u03bcL, with the following composition: 1x PCR buffer, 2.5 mM MgCl2<\/sub>, 2.0 mM each deoxynucleotide triphosphates, 10 pM each primers for detection and 1 unit DNA polymerase.
\nDNA was added in an amount of 5.0 \u03bcL (100-150 ng). Studies were performed on a thermal cycler 2720 (Applied Biosystems, USA) with the temperature profile given in the relevant literature source. The nucleotide sequences and other characteristics of the primers used in the study are given in Table 1.<\/p>\n

\"\"
Table 1<\/strong>. Characteristics of primers that were used in the studies.<\/figcaption><\/figure>\n

The amplification products were separated in 1.5% agarose gel.<\/p>\n

Molecular genetic studies were performed to identify the presence of the genes toxA<\/em>, which is responsible for producing a dermonecrotic toxin, and ptfA<\/em>, which encodes production of the products that assemble to form type 4 fimbriae on the bacterial surface and are responsible for biofilm formation (Devi et al<\/em>., 2018; Gong et al<\/em>., 2020).<\/p>\n

<\/a> ▲<\/a><\/p>\n

Results<\/h2>\n
\n

This paper presents the results of a study of 11 isolates of Pasteurella isolated from pathological material from 24 rabbits and five piglets, and smears from 12 clinically healthy dogs and nine cats (Table 2): six isolates (54.5%) were from rabbits, two isolate (18.1%) from dogs, two (18.1%) from cats, and one (9.1%) from pigs.<\/p>\n

\"\"
Table 2<\/strong>. Results of determination of toxA<\/em> gene, virulence, antibiotic resistance, ptfA<\/em> biofilm gene in isolates of P. multocida<\/em>, and the results of phenotypic determination of biofilm formation (by crystal violet and Congo red staining).
* CFU \u2013 colony forming units.<\/figcaption><\/figure>\n

The virulence factor of the toxA<\/em> gene (Table 2, Figure 1), which is responsible for the formation of a dermatonecrotic toxin, was present in 72.7% of cases in P. multocida<\/em> strains P5, P15, P16, P17, P50, P97, P99, and P99p.<\/p>\n

\"\"
Figure 1<\/strong>. P. multocida<\/em> polymerase chain reaction for the detection of the toxA gene (846 bp) of P. multocida<\/em>. Lane A and I \u2013 marker lanes; lanes B, H, M: samples negative for toxA<\/em> gene; Lanes C, D, E, F, G, J, K, L: samples positive for toxA<\/em> gene; Lane N \u2013 positive control for toxA<\/em> gene; Lane O \u2013 negative control for toxA<\/em> gene.<\/figcaption><\/figure>\n

The isolate P. multocida<\/em> P50 had the highest virulence. At 6 hours after infection, the death of 100% of animals was recorded in all experimental groups, LD100<\/sub> – 0.5 \u00d7 101<\/sup> CFU (Table 2). LD100<\/sub> for the isolates of P. multocida<\/em> P5, P15, P16, P99 and P99p was 0.5 \u00d7 104<\/sup> CFU, the death of experimental animals was recorded 14\u201318 hours after infection.
\nLD100<\/sub> for isolates of P. multocida<\/em> P2, P17, PC, P97 and P69p was 0.5 \u00d7 106<\/sup> CFU, and the death of experimental animals was recorded 24 hours after infection.<\/p>\n

The sensitivity of P. multocida<\/em> isolates to the antibacterial drugs penicillin, cephalosporin, fluoroquinolol and tetracycline groups was determined. The susceptibility to P. multocida<\/em> antibiotics are shown in Table 3: 54.5% of isolates were resistant to benzylpenicillin, 45.4% to cefotaxime, 18.2% to ampicillin, and none of the studied isolates showed resistance to amoxicillin.<\/p>\n

Examining the group of fluoroquinolones showed that 27.3% of isolates of P. multocida<\/em> were resistant to ciprofloxacin and 54.4% to levofloxacin.
\nResistance to nalidixic acid (screening) was demonstrated at 18.2% for each drug.
\nScreening for fluoroquinolone resistance for nalidixic acid sensitivity was positive for only four (36.4%) P. multocida<\/em> isolates (P2, P50, P99, P69p).<\/p>\n

A study of the tetracycline group found that 54.5% of isolates were resistant to doxycycline and 18.2% of P. multocida<\/em> isolates were resistant to tetracycline.<\/p>\n

Isolate P5 showed resistance to the group of penicillins (benzylpenicillin, ampicillin), cephalosporins (cefotaxime), fluoroquinolones (levofloxacin), indicating that this isolate is multidrug-resistant (MDR). The isolate \u0420\u0421 was also included in this category of resistance, showing resistance to the group of penicillins (benzylpenicillin), cephalosporins (cefotaxime), fluoroquinolones (levofloxacin, ciprofloxacin), tetracyclines (doxycycline). Isolate P50 with extreme multidrug-resistance (XDR) showed resistance to the group of penicillins (benzylpenicillin, ampicillin), cephalosporins (cefotaxime), fluoroquinolones (levofloxacin, ciprofloxacin), tetracyclines (doxycycline, tetracycline), and nalidixic acid.<\/p>\n

\"\"
Figure 2<\/strong>. Biofilm formation of P. multocida<\/em> stained with crystal violet (top) and Congo-red (bottom), dishes on the left \u2013 control.<\/figcaption><\/figure>\n

The density of the biofilm was determined by staining methods with crystal violet and Congo red (Table 2, Figure 2).<\/p>\n

The results showed that these staining methods provide close in value results, deviations of the optical density results were registered in the range of 2.2\u201317.2%, which did not exceed 15%.<\/p>\n

Therefore, in the study of the phenotypic biofilm formation, staining with Congo-red did not differ from the method with crystalline violet.<\/p>\n

In the study of the phenotypic biofilm formation using crystalline violet and Congo-red staining methods It was found that one isolate (9.1%) formed a high-density biofilm (\u03bb-1.7). The medium-density biofilm was formed by three (27.3%) isolates P. multocida<\/em>: P5 \u2013 \u03bb 0.8, P16 \u2013 \u03bb 0.5, P97 \u2013 \u03bb 0.5. The low-density biofilm (\u03bb 0.13 \u2013 0.3) was formed by 63.6% of the studied isolates P. multocida<\/em>.<\/p>\n

In three isolates (isolated from rabbits and pigs, LD100<\/sub> 0.5×104<\/sup>\u20130.5×106<\/sup> CFU) the density of the biofilm ranged from \u03bb 0.1344\u20130.1801. In four isolates, the optical density of the biofilm in vitro<\/em> was \u03bb 0.2162\u20130.3082. These isolates were isolated from rabbits (1), dogs (1) and cats (2), and virulence was 0.5×104<\/sup>\u20130.5×106<\/sup> CFU. In two isolates, the optical density of the biofilm was \u03bb 0.5053\u20130.5505. These isolates were isolated from rabbit and dog, and LD100<\/sub> was 0.5×104<\/sup> and 0.5×106<\/sup> CFU, respectively. One culture of P. multocida<\/em> isolated from rabbits with a virulence of LD100<\/sub> 0.5×104<\/sup> formed a biofilm in vitro<\/em> with an optical density \u03bb 0.8593. The isolate with the highest virulence LD100<\/sub> 0.5×101<\/sup> formed a biofilm with the highest optical density \u03bb 1.7893.<\/p>\n

Finally, it should be noted that all isolates possessed the ptfA<\/em> gene (Table 3, Figure 3), which is responsible for the formation of special adhesives (type 4 pili).<\/p>\n

\"\"
Table 3<\/strong>. Sensitivity testing to antibiotics in isolated isolates of P. multocida<\/em>, according to the EUCAST recommendations, (mm).<\/figcaption><\/figure>\n
\"\"
Figure 3<\/strong>. P. multocida<\/em> polymerase chain reaction for the detection of the ptfA<\/em> gene (435 bp) of P. multocida<\/em>. Lane A and I – marker lanes; Lanes D, E, F, G, H, J, K, L, M, N, O: samples positive for ptfA<\/em> gene; Lane C – positive control for ptfA<\/em> gene; Lane B \u2013 negative control for ptfA<\/em> gene.<\/figcaption><\/figure>\n

<\/a> ▲<\/a><\/p>\n

Discussion<\/h2>\n
\n

In this study, 11 isolates of P. multocida<\/em> were used. P. multocida<\/em> isolates were pathogenic for white mice, LD100<\/sub> ranged from 0.5×101<\/sup> to 0.5×106<\/sup> CFU. The virulence index LD100<\/sub> 0.5×106<\/sup> CFU was found in isolates from rabbits (2), dogs (2), pigs (1). The virulence index LD100<\/sub> 0.5×104<\/sup> CFU was found in five isolates from rabbits (3) and cats (2), while the virulence index LD100<\/sub> 0.5×101<\/sup> CFU was found in the isolate from rabbit.<\/p>\n

Among the studied strains, eight (72.7%) were toxA<\/em> positive (formed dermonecrotic toxin), while five (45.4%) had a complete set of phenotypic (biofilm formation, lethality) and genotypic (toxA<\/em>) traits. Three strains (27.3%) showed significant signs of multidrug resistance. All studied isolates possessed the gene ptfA<\/em> that encodes the formation of fimbriae type 4, which participates in the formation of the biofilm. Virulence in toxA<\/em>-negative P. multocida<\/em> was lower compared to toxA<\/em>-positive isolates.<\/p>\n

It should be noted that the determination of biofilm density obtained by staining crystal violet and Congo red did not show a deviation of optical density results exceeding 15%.<\/p>\n

As a result of determining the sensitivity of P. multocida<\/em> isolates to antibacterial drugs, it was found that the cultures P5 and PC showed signs of multiple drug resistance MDR, and culture P50 exhibited extensive drug resistance (XDR).<\/p>\n

The ability to form a biofilm and the presence of the gene for the adhesive ability of cultures was found in all strains of P. multocida<\/em>, though there was a tendency to increase the optical density of the biofilm at a higher strain virulence.
\nThis indicates the ability to form a biofilm in P. multocida<\/em> as another possible sign of bacterial virulence.<\/p>\n

It should also be noted that two strains of P. multocida<\/em> that showed signs of polyresistance (P5 and P50), were isolated from rabbits from a single farm, and this indicate that the antibiotic-resistant strains of P. multocida<\/em> are confined to this farm. It is also important to distinguish MDR-Pasteurella from a dog as a companion animal, which can make this a medical issue, since P. multocida<\/em> can cause pathological processes in both animals and humans.<\/p>\n

The results indicate a possible relationship between the level of P. multocida<\/em> virulence and the density of the formed biofilm; we believe that an in-depth study of this issue will be the subject of our future research.<\/p>\n

It should also be noted that the phenotypic manifestation of the ability to form biofilms in this study did not coincide with the results of the relevant genetic marker detection. Indirectly, this fact indicates the need for in-depth study of this phenomenon to identify previously unknown genetic loci that determine the phenotypic manifestation of resistance and the ability to form biofilms, which is important for understanding the pathogenesis of pasteurellosis and their effective detection. In addition, data on the prevalence of virulence factors will be the scientific basis for improving the specific prevention of animal pasteurellosis.<\/p>\n

<\/a> ▲<\/a><\/p>\n

Conclusion<\/h2>\n
\n

As a result of the study of 50 samples of biological material from animals, 11 isolates of P. multocida<\/em> were isolated: six from rabbits, two from dogs, two from cats and one from pigs.<\/p>\n

Isolated isolates of P. multocida<\/em> were sensitive to amoxicillin; one isolate was assigned to XDR, and two isolates to MDR. Screening of susceptibility of P. multocida<\/em> isolates to fluoroquinolones by the disc-diffusion method, sensitivity to nalidixic acid did not coincide in 63.6% of cases, which gives grounds for further studies.<\/p>\n

The isolated cultures were pathogenic to white mice. The LD100<\/sub> of isolate P50 was 0.5×101<\/sup> CFU in the lightning course of the disease. In five isolates, LD100<\/sub> was 0.5×104<\/sup> CFU, and in five others was 0.5×106<\/sup> CFU.
\nNo association was established between virulence and the species of animals from which P. multocida<\/em> has been isolated. The toxA<\/em> gene was detected in eight cultures with LD100<\/sub> 0.5×101<\/sup>\u20130.5×106<\/sup> CFU isolated from rabbits, dogs and cats; in three isolates with LD100<\/sub> 0.5×106<\/sup> CFU this gene was not detected.<\/p>\n

In the study of phenotypic biofilm formation using Congo-red staining, the results did not differ from the method with crystalline violet. The genome of all studied isolates of P. multocida<\/em> contained the ptfA<\/em> gene; the studied cultures in vitro formed a biofilm of varying density.
\nThe culture with the highest virulence (0.5×101<\/sup> CFU) and extreme resistance to antibiotics formed a biofilm of the highest density.<\/p>\n

From this study, it can be concluded that the toxA<\/em> gene is an important marker gene for defining the pathogenic potential of P. multocida<\/em> strains in various animals. However, other virulence genes are also found to be widely distributed among pathogenic strains of P. multocida<\/em>.
\nThe association of the gene in the biofilm-producing mechanism needs further evaluation. Further research is also needed to identify the relationships between pathogens among P. multocida<\/em> isolates from different species of animals and humans.<\/p>\n

<\/a>
\nReferences<\/strong> [… show]<\/span><\/a><\/span><\/p>\n

  ▲<\/a><\/p>\n

1.\tBRUCHMANN, S., T. FELTWELL, J. PARKHILL and F. L SHORT (2021): Identifying virulence determinants of multidrug-resistant Klebsiella pneumoniae in Galleria mellonella. Pathog. Dis. 79, ftab009, 10.1093\/femspd\/ftab009
\n2.\tCOENYE, T., and H. J. NELIS (2010): In vitro and in vivo model systems to study microbial biofilm formation. J. Microbiol. Methods 83, 89-105. 10.1016\/j. mimet.2010.08.018
\n3.\tCURTIS, P. E. (1985): In: Pasteurella multocida. Isolation and Identification of Microorganisms of Medical and Veterinary Importance. Collins, C. H, Grange, J. M, eds. London: Academic Press.
\n4.\tDEVI, L. B., D. P. BORA, S. K. DAS, R. K. SHARMA, S. MUKHERJEE and R. A. HAZARIKA (2018): Virulence gene profiling of porcine Pasteurella multocida isolates of Assam. Vet. World. 11, 348-354.
\n5.\tDONLAN, R. M. and J. W. COSTERTON (2002): Biofilms: survival mechanisms of clinically relevant microorganisms. Clin. Microbiol. Rev. 15, 167-193.
\n6.\tDOUGHTY, S. W., C. G. RUFFOLO and B. ADLER (2000): The type 4 fimbrial subunit gene of pasteurella multocida. Vet. Microbiol. 7279-7290. 10.1016\/S0378-1135(99)00189-3
\n7.\tEUCAST. The european committee on antimicrobial susceptibility testing (2021): Available from: http:\/\/ www.eucast.org.
\n8.\tEWERS, C., A. LUBKE-BECKER, A. BETHE, et al. (2000): Virulence genotype of Pasteurella multocida strains isolated from different hosts with various disease status. Vet. Microbiol. 114, 304-317.
\n9.\tGONG, Q., Y. G. PENG, M. F. NIU, C. L. QIN (2020): The immune enhancement ability of inulin on ptfA gene DNA vaccine of avian Pasteurella multocida. Poultry Sci. 99, 3015-3019. 10.1016\/j.psj.2020.03.006
\n10.\tGUAN, L, Y. XUE, W. DING and Z. ZHAO (2019): Biosynthesis and regulation mechanisms of the Pasteurella multocida capsule. Res. Vet. Sci. 127, 82-90. 10.1016\/j.rvsc.2019.10.011.
\n11.\tGUAN, L. J., J. J. SONG, Y. XUE, X. AI, Z. J. LIU, L. F. SI, M. Y. LI and Z. Q. ZHAO (2021): Immune Protective Efficacy of China\u2019s Traditional Inactivated and Attenuated Vaccines against the Prevalent Strains of Pasteurella multocida in Mice. Vaccines (Basel). 9, 1155. 10.3390\/vaccines9101155
\n12.\tGUAN, L., L. ZHANG, Y. XUE, J. YANG and Z. ZHAO (2020): Molecular pathogenesis of the hyaluronic acid capsule of Pasteurella multocida. Microb. Pathog. 149, 104380. 10.1016\/j. micpath.2020.104380.
\n13.\tJAMAL, M, W. AHMAD, S. ANDLEEB, F. JALIL, M. IMRAN, M. A. NAWAZ, T. HUSSAIN, N. ALI, M. RAFIQ and M. A. KAMIL (2018): Bacterial biofilm and associated infections. J. Chin. Med. Assoc. 81, 7-11. 10.1016\/j.jcma.2017.07.012.
\n14.\tKUKHTYN, M. and N. KRUSHELNYTSKA (2014): Forming of biofilms of microorganisms obtained from milking equipment. Anim. Biol. 16, 95-103.
\n15.\tMAGIORAKOS, A.-P., A. SRINIVASAN, R. B. CAREY, et al. (2012): Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin. Microbiol. Infect. 18, 268-281. 10.1111\/j.1469- 0691.2011.03570.x.
\n16.\tPENG, Z., H. WANG, W. LIANG, Y. CHEN, X. TANG, H. CHEN and B. WU (2018): A capsule\/ lipopolysaccharide\/MLST genotype D\/L6\/ST11 of Pasteurella multocida is likely to be strongly associated with swine respiratory disease in China. Arch. Microbiol. 200, 107-118. 10.1007\/s00203-017-1421-y.
\n17.\tPETRUZZI, B., R. A. DALLOUL, T. LEROITH, N. P. EVANS, F. W. PIERSON and T. J. INZANA (2018): Biofilm formation and avian immune response following experimental acute and chronic avian cholera due to Pasteurella multocida. Vet Microbiol. 222, 114-123. 10.1016\/j.vetmic.2018.07.005
\n18.\tPETRUZZI, B., A. DICKERMAN, K. LAHMERS, W. SCARRATT and T. INZANA (2020): Polymicrobial Biofilm Interaction Between Histophilus somni and Pasteurella multocida Front. Microbiol. 11, 1561. 10.3389\/fmicb.2020.01561
\n19.\tRAJAGOPAL, R., G. K. NAIR, M. MINI, L. JOSEPH, M. R. SASEENDRANATH and K. JOHN (2013): Biofilm formation of Pasteurella multocida on bentonite clay. Iran J. Microbiol. 5, 120-125.
\n20.\tSTEEN, J. A., J. A. STEEN, P. HARRISON, T. SEEMANN, I. WILKIE, M. HARPER, B. ADLER and J. D. BOYCE (2010): Fis is essential for capsule production in Pasteurella multocida and regulates expression of other important virulence factors. PloS Pathog. 6, e1000750 10.1371\/journal.ppat.1000750.
\n21.\tWELIN, J., J. C. WILKINS, D. BEIGHTON and G. SVENSATER (2004): Protein expression by Streptococcus mutans during initial stage of biofilm formation. Appl. Environ. Microbiol. 70, 3736-3741.
\n<\/em><\/p>\n<\/div>\n

<\/a> ▲<\/a><\/p>\n

\n

Studija znakova patogenosti kod Pasteurella multocida<\/em>, izolirane iz \u017eivotinja razli\u010ditih vrsta<\/h2>\n
\n
Serhii BOIANOVSKIY<\/strong>, Valerii USHKALOV<\/strong>, Lilia VYGOVSKA<\/strong>, Tatyana MAZUR<\/strong>, Liudmyla ISHCHENKO<\/strong>, National University of Life and Environmental Sciences of Ukraine 15 Heroyiv Oborony str., Kyiv, Ukraine; Kateryna RUDNIEVA<\/strong>, Kyiv Regional Clinical Hospital, 1 Baggovutivska str., Kyiv, Ukraine; Artem USHKALOV<\/strong>, Main administration of state service of Ukraine on food safety and consumer protection in Kharkiv reg. Ukraine; Volodymyr MELNYK<\/strong>, National University of Life and Environmental Sciences of Ukraine 15 Heroyiv Oborony str., Kyiv, Ukraine<\/div>\n
\n

Znatan broj mikroorganizama u prirodnim i umjetnim okru\u017eenjima postoji u obliku strukturirane formacije \u2013 biofilma i ta se formacija mo\u017ee vezati i na odre\u0111enu povr\u0161inu, posebice na epitel. Mogu\u0107nost formiranja sli\u010dne strukture zamije\u0107ena je kod Pasteurella multocida<\/em>, koja je uzro\u010dnik antropozoonoza, a poga\u0111a i ljude, ku\u0107ne ljubimce, ptice i doma\u0107e i divlje \u017eivotinje. Spektar patogenih u\u010dinaka P. multocida<\/em> prili\u010dno je \u0161irok i povezan je s razvojem respiratorne i multisistemske bolesti, bakterijemije i drugih manifestacija. Pravovremeno otkrivanje P. multocida<\/em> i lije\u010denje bolesti koje ova bakterija prouzro\u010di u \u017eivotinja na farmi i doma\u0107ih \u017eivotinja va\u017eno je za ograni\u010denje ekonomskih gubitaka i sigurnost dru\u0161tva. Glavni je cilj ove studije bio utvrditi patogenost P. multocida<\/em>, sposobnost formiranja biofilma, otpornost na antibiotike, ali i identificirati gene odgovorne za formiranje dermonekroti\u010dnog toksina, odnosno formiranje biofilma. Ovaj rad predstavlja rezultate studije 11 izolata P. multocida<\/em>: 6 izolata (54,5 %) izoliranih iz ze\u010deva, 2 (18,2 %) iz pasa, 2 (18,2 %) iz ma\u010daka te 1 izolat izoliran iz svinja (9,2 %). U 100 % prou\u010davanih izolata, otkriven je gen (ptfA<\/em>) koji kodira formiranje fimbrija tipa 4 i sudjeluje u formiranju biofilma. Ispitane su kulture in vitro<\/em> formirale biofilm razli\u010ditih gusto\u0107a. U genomu 8 ispitanih izolata (72,7 %) otkrivena je prisutnost toxA<\/em> gena (koji omogu\u0107uje formiranje dermonekroti\u010dnog toksina). 45,4 % ispitanih izolata pokazalo je cijeli set prou\u010davanih znakova patogenosti – fenotipskih (formiranje biofilma, smrtnost za laboratorijske \u017eivotinje) karakteristika i genotipskih (prisutnost toxA<\/em>, ptfA<\/em>) karakteristika. Tri izolata (27,3 %) pokazala su otpornost na vi\u0161e lijekova. Otkriveno je da je kod toxA<\/em>-negativnih izolata P. multocida<\/em> virulencija bila ni\u017ea u usporedbi s toxA<\/em>-pozitivnim izolatima. Kultura s najvi\u0161om virulentno\u0161\u0107u (0,5 x 101<\/sup> CFU) i ekstremnom otporno\u0161\u0107u na antibiotike formirala je biofilm najve\u0107e gusto\u0107e. Asocijacija gena u mehanizmu proizvodnje biofilma zahtijeva dodatnu procjenu, a potrebno je i dodatno istra\u017eivanje za identifikaciju odnosa izme\u0111u patogena me\u0111u izolatima P. multocida<\/em> izoliranima iz ljudi i razli\u010ditih vrsta \u017eivotinja.<\/p>\n

Klju\u010dne rije\u010di:<\/strong> biofilm, Pasteurella multocida, otpornost na antibiotike, toxA, ptfA<\/em><\/p><\/blockquote>\n","protected":false},"excerpt":{"rendered":"

S. Boianovskiy, V. Ushkalov*, L. Vygovska, T. Mazur, L. Ishchenko, K. Rudnieva, A. Ushkalov and V. Melnyk Serhii BOIANOVSKIY, Valerii<\/p>\n","protected":false},"author":8,"featured_media":0,"menu_order":6,"comment_status":"closed","ping_status":"open","template":"","format":"standard","meta":{"footnotes":""},"categories":[21],"tags":[1602,1604,1603,1606,1605],"issuem_issue":[1590],"ppma_author":[1607,1465,1608,1609,1610,1611,1612,1613],"class_list":["post-5448","article","type-article","status-publish","format-standard","hentry","category-original-scientific-articles","tag-biofilm","tag-otpornost-na-antibiotike","tag-pasteurella-multocida","tag-ptfa","tag-toxa","issuem_issue-veterinarska-stanica-54-3"],"yoast_head":"\nSigns of pathogenicity by Pasteurella multocida in different species of animals - CROATIAN VETERINARY JOURNAL<\/title>\n<meta name=\"description\" content=\"A significant number of microorganisms in natural and artificial environments exist in a structured formation \u2013 biofilm. This formation attaches to a certain surface, particularly the epithelium. The ability to form a similar structure has been observed in Pasteurella multocida, the causative agent of anthropozoonoses that affect domestic and wild animals, birds, companion animals and humans. The spectrum of pathogenetic action of P. multocida is wide and associated with the development of respiratory and multisystemic pathology, bacteraemia and other manifestations.\" \/>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals\" \/>\n<meta property=\"og:locale\" content=\"en_GB\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Signs of pathogenicity by Pasteurella multocida in different species of animals - CROATIAN VETERINARY JOURNAL\" \/>\n<meta property=\"og:description\" content=\"A significant number of microorganisms in natural and artificial environments exist in a structured formation \u2013 biofilm. This formation attaches to a certain surface, particularly the epithelium. The ability to form a similar structure has been observed in Pasteurella multocida, the causative agent of anthropozoonoses that affect domestic and wild animals, birds, companion animals and humans. The spectrum of pathogenetic action of P. multocida is wide and associated with the development of respiratory and multisystemic pathology, bacteraemia and other manifestations.\" \/>\n<meta property=\"og:url\" content=\"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals\" \/>\n<meta property=\"og:site_name\" content=\"CROATIAN VETERINARY JOURNAL\" \/>\n<meta property=\"article:publisher\" content=\"https:\/\/www.facebook.com\/pages\/Hrvatski%20Veterinarski%20Institut\/291017291058567\/\" \/>\n<meta property=\"og:image\" content=\"https:\/\/veterinarska-stanica-journal.hr\/wp-content\/uploads\/2022\/10\/SerhiiBOIANOVSKIY.jpg\" \/>\n<meta name=\"twitter:card\" content=\"summary_large_image\" \/>\n<meta name=\"twitter:label1\" content=\"Estimated reading time\" \/>\n\t<meta name=\"twitter:data1\" content=\"27 minutes\" \/>\n<script type=\"application\/ld+json\" class=\"yoast-schema-graph\">{\"@context\":\"https:\/\/schema.org\",\"@graph\":[{\"@type\":\"WebPage\",\"@id\":\"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals\",\"url\":\"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals\",\"name\":\"Signs of pathogenicity by Pasteurella multocida in different species of animals - CROATIAN VETERINARY JOURNAL\",\"isPartOf\":{\"@id\":\"https:\/\/journal.h3s.org\/#website\"},\"primaryImageOfPage\":{\"@id\":\"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals#primaryimage\"},\"image\":{\"@id\":\"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals#primaryimage\"},\"thumbnailUrl\":\"https:\/\/veterinarska-stanica-journal.hr\/wp-content\/uploads\/2022\/10\/SerhiiBOIANOVSKIY.jpg\",\"datePublished\":\"2022-10-13T18:13:51+00:00\",\"description\":\"A significant number of microorganisms in natural and artificial environments exist in a structured formation \u2013 biofilm. This formation attaches to a certain surface, particularly the epithelium. The ability to form a similar structure has been observed in Pasteurella multocida, the causative agent of anthropozoonoses that affect domestic and wild animals, birds, companion animals and humans. The spectrum of pathogenetic action of P. multocida is wide and associated with the development of respiratory and multisystemic pathology, bacteraemia and other manifestations.\",\"breadcrumb\":{\"@id\":\"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals#breadcrumb\"},\"inLanguage\":\"en-GB\",\"potentialAction\":[{\"@type\":\"ReadAction\",\"target\":[\"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals\"]}]},{\"@type\":\"ImageObject\",\"inLanguage\":\"en-GB\",\"@id\":\"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals#primaryimage\",\"url\":\"https:\/\/veterinarska-stanica-journal.hr\/wp-content\/uploads\/2022\/10\/SerhiiBOIANOVSKIY.jpg\",\"contentUrl\":\"https:\/\/veterinarska-stanica-journal.hr\/wp-content\/uploads\/2022\/10\/SerhiiBOIANOVSKIY.jpg\"},{\"@type\":\"BreadcrumbList\",\"@id\":\"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals#breadcrumb\",\"itemListElement\":[{\"@type\":\"ListItem\",\"position\":1,\"name\":\"Home\",\"item\":\"https:\/\/journal.h3s.org\/\"},{\"@type\":\"ListItem\",\"position\":2,\"name\":\"Articles\",\"item\":\"https:\/\/journal.h3s.org\/?post_type=article\"},{\"@type\":\"ListItem\",\"position\":3,\"name\":\"Signs of pathogenicity by Pasteurella multocida in different species of animals\"}]},{\"@type\":\"WebSite\",\"@id\":\"https:\/\/journal.h3s.org\/#website\",\"url\":\"https:\/\/journal.h3s.org\/\",\"name\":\"VETERINARSKA STANICA\",\"description\":\"Journal of Croatian Veterinary Institute\",\"publisher\":{\"@id\":\"https:\/\/journal.h3s.org\/#organization\"},\"potentialAction\":[{\"@type\":\"SearchAction\",\"target\":{\"@type\":\"EntryPoint\",\"urlTemplate\":\"https:\/\/journal.h3s.org\/?s={search_term_string}\"},\"query-input\":{\"@type\":\"PropertyValueSpecification\",\"valueRequired\":true,\"valueName\":\"search_term_string\"}}],\"inLanguage\":\"en-GB\"},{\"@type\":\"Organization\",\"@id\":\"https:\/\/journal.h3s.org\/#organization\",\"name\":\"Veterinarska stanica\",\"url\":\"https:\/\/journal.h3s.org\/\",\"logo\":{\"@type\":\"ImageObject\",\"inLanguage\":\"en-GB\",\"@id\":\"https:\/\/journal.h3s.org\/#\/schema\/logo\/image\/\",\"url\":\"https:\/\/veterinarska-stanica-journal.hr\/wp-content\/uploads\/2021\/03\/veterinarska-stanica-casopis-hvi-728.png\",\"contentUrl\":\"https:\/\/veterinarska-stanica-journal.hr\/wp-content\/uploads\/2021\/03\/veterinarska-stanica-casopis-hvi-728.png\",\"width\":728,\"height\":90,\"caption\":\"Veterinarska stanica\"},\"image\":{\"@id\":\"https:\/\/journal.h3s.org\/#\/schema\/logo\/image\/\"},\"sameAs\":[\"https:\/\/www.facebook.com\/pages\/Hrvatski Veterinarski Institut\/291017291058567\/\",\"https:\/\/www.linkedin.com\/company\/croatian-veterinary-institute\/\",\"https:\/\/www.youtube.com\/watch?v=BFn739WHdcU&amp;amp;t=2s\"]}]}<\/script>\n<!-- \/ Yoast SEO plugin. -->","yoast_head_json":{"title":"Signs of pathogenicity by Pasteurella multocida in different species of animals - CROATIAN VETERINARY JOURNAL","description":"A significant number of microorganisms in natural and artificial environments exist in a structured formation \u2013 biofilm. This formation attaches to a certain surface, particularly the epithelium. The ability to form a similar structure has been observed in Pasteurella multocida, the causative agent of anthropozoonoses that affect domestic and wild animals, birds, companion animals and humans. The spectrum of pathogenetic action of P. multocida is wide and associated with the development of respiratory and multisystemic pathology, bacteraemia and other manifestations.","robots":{"index":"index","follow":"follow","max-snippet":"max-snippet:-1","max-image-preview":"max-image-preview:large","max-video-preview":"max-video-preview:-1"},"canonical":"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals","og_locale":"en_GB","og_type":"article","og_title":"Signs of pathogenicity by Pasteurella multocida in different species of animals - CROATIAN VETERINARY JOURNAL","og_description":"A significant number of microorganisms in natural and artificial environments exist in a structured formation \u2013 biofilm. This formation attaches to a certain surface, particularly the epithelium. The ability to form a similar structure has been observed in Pasteurella multocida, the causative agent of anthropozoonoses that affect domestic and wild animals, birds, companion animals and humans. The spectrum of pathogenetic action of P. multocida is wide and associated with the development of respiratory and multisystemic pathology, bacteraemia and other manifestations.","og_url":"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals","og_site_name":"CROATIAN VETERINARY JOURNAL","article_publisher":"https:\/\/www.facebook.com\/pages\/Hrvatski%20Veterinarski%20Institut\/291017291058567\/","og_image":[{"url":"https:\/\/veterinarska-stanica-journal.hr\/wp-content\/uploads\/2022\/10\/SerhiiBOIANOVSKIY.jpg","type":"","width":"","height":""}],"twitter_card":"summary_large_image","twitter_misc":{"Estimated reading time":"27 minutes"},"schema":{"@context":"https:\/\/schema.org","@graph":[{"@type":"WebPage","@id":"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals","url":"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals","name":"Signs of pathogenicity by Pasteurella multocida in different species of animals - CROATIAN VETERINARY JOURNAL","isPartOf":{"@id":"https:\/\/journal.h3s.org\/#website"},"primaryImageOfPage":{"@id":"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals#primaryimage"},"image":{"@id":"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals#primaryimage"},"thumbnailUrl":"https:\/\/veterinarska-stanica-journal.hr\/wp-content\/uploads\/2022\/10\/SerhiiBOIANOVSKIY.jpg","datePublished":"2022-10-13T18:13:51+00:00","description":"A significant number of microorganisms in natural and artificial environments exist in a structured formation \u2013 biofilm. This formation attaches to a certain surface, particularly the epithelium. The ability to form a similar structure has been observed in Pasteurella multocida, the causative agent of anthropozoonoses that affect domestic and wild animals, birds, companion animals and humans. The spectrum of pathogenetic action of P. multocida is wide and associated with the development of respiratory and multisystemic pathology, bacteraemia and other manifestations.","breadcrumb":{"@id":"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals#breadcrumb"},"inLanguage":"en-GB","potentialAction":[{"@type":"ReadAction","target":["https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals"]}]},{"@type":"ImageObject","inLanguage":"en-GB","@id":"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals#primaryimage","url":"https:\/\/veterinarska-stanica-journal.hr\/wp-content\/uploads\/2022\/10\/SerhiiBOIANOVSKIY.jpg","contentUrl":"https:\/\/veterinarska-stanica-journal.hr\/wp-content\/uploads\/2022\/10\/SerhiiBOIANOVSKIY.jpg"},{"@type":"BreadcrumbList","@id":"https:\/\/journal.h3s.org\/?article=signs-of-pathogenicity-by-pasteurella-multocida-in-different-species-of-animals#breadcrumb","itemListElement":[{"@type":"ListItem","position":1,"name":"Home","item":"https:\/\/journal.h3s.org\/"},{"@type":"ListItem","position":2,"name":"Articles","item":"https:\/\/journal.h3s.org\/?post_type=article"},{"@type":"ListItem","position":3,"name":"Signs of pathogenicity by Pasteurella multocida in different species of animals"}]},{"@type":"WebSite","@id":"https:\/\/journal.h3s.org\/#website","url":"https:\/\/journal.h3s.org\/","name":"VETERINARSKA STANICA","description":"Journal of Croatian Veterinary Institute","publisher":{"@id":"https:\/\/journal.h3s.org\/#organization"},"potentialAction":[{"@type":"SearchAction","target":{"@type":"EntryPoint","urlTemplate":"https:\/\/journal.h3s.org\/?s={search_term_string}"},"query-input":{"@type":"PropertyValueSpecification","valueRequired":true,"valueName":"search_term_string"}}],"inLanguage":"en-GB"},{"@type":"Organization","@id":"https:\/\/journal.h3s.org\/#organization","name":"Veterinarska stanica","url":"https:\/\/journal.h3s.org\/","logo":{"@type":"ImageObject","inLanguage":"en-GB","@id":"https:\/\/journal.h3s.org\/#\/schema\/logo\/image\/","url":"https:\/\/veterinarska-stanica-journal.hr\/wp-content\/uploads\/2021\/03\/veterinarska-stanica-casopis-hvi-728.png","contentUrl":"https:\/\/veterinarska-stanica-journal.hr\/wp-content\/uploads\/2021\/03\/veterinarska-stanica-casopis-hvi-728.png","width":728,"height":90,"caption":"Veterinarska stanica"},"image":{"@id":"https:\/\/journal.h3s.org\/#\/schema\/logo\/image\/"},"sameAs":["https:\/\/www.facebook.com\/pages\/Hrvatski Veterinarski Institut\/291017291058567\/","https:\/\/www.linkedin.com\/company\/croatian-veterinary-institute\/","https:\/\/www.youtube.com\/watch?v=BFn739WHdcU&amp;amp;t=2s"]}]}},"_links":{"self":[{"href":"https:\/\/journal.h3s.org\/index.php?rest_route=\/wp\/v2\/article\/5448","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/journal.h3s.org\/index.php?rest_route=\/wp\/v2\/article"}],"about":[{"href":"https:\/\/journal.h3s.org\/index.php?rest_route=\/wp\/v2\/types\/article"}],"author":[{"embeddable":true,"href":"https:\/\/journal.h3s.org\/index.php?rest_route=\/wp\/v2\/users\/8"}],"replies":[{"embeddable":true,"href":"https:\/\/journal.h3s.org\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=5448"}],"version-history":[{"count":9,"href":"https:\/\/journal.h3s.org\/index.php?rest_route=\/wp\/v2\/article\/5448\/revisions"}],"predecessor-version":[{"id":5468,"href":"https:\/\/journal.h3s.org\/index.php?rest_route=\/wp\/v2\/article\/5448\/revisions\/5468"}],"wp:attachment":[{"href":"https:\/\/journal.h3s.org\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=5448"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/journal.h3s.org\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=5448"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/journal.h3s.org\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=5448"},{"taxonomy":"issuem_issue","embeddable":true,"href":"https:\/\/journal.h3s.org\/index.php?rest_route=%2Fwp%2Fv2%2Fissuem_issue&post=5448"},{"taxonomy":"author","embeddable":true,"href":"https:\/\/journal.h3s.org\/index.php?rest_route=%2Fwp%2Fv2%2Fppma_author&post=5448"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}