Mass spectrometry-based identification of 26 Pasteurella species and in vitro antimicrobial susceptibility pattern of isolates recovered from diseased domestic catsABSTRACT:
Pasteurella species are well-known opportunistic bacteria that inhabit the microbiota of the oral cavity and upper respiratory tract of cats and have been related to a set of pet-associated diseases in addition to humans. Most studies involving feline pasteurellosis have been described as case reports and species identification based on classic phenotypic methods. In turn, a lack of comprehensive studies involving a great number of cats with pasteurellosis has been described, especially where diagnosis at the species level has been performed by molecular-based methods. In this scenario, we investigated the molecular identification of Pasteurella species isolated from 26 diseased domestic cats (i.e., cutaneous abscesses, pneumonia, conjunctivitis, open wounds, urinary tract infections, pleural effusion, pyometra, and infection secondary to neoplasia) based on proteomic diagnosis, using mass spectrometry (matrix-assisted laser desorption ionization time-of-flight mass spectrometry - MALDI-TOF MS). The in vitro antimicrobial susceptibility patterns of isolates and selected epidemiological data (with emphasis on the outcome) were assessed as well. MALDI-TOF MS identified predominantly P. multocida (23/26=88.5%), followed by P. dagmatis (2/26=7.7%) and P. canis (1/26=3.8%). The isolates revealed 100% susceptibility to beta-lactams (amoxicillin/clavulanic acid, ampicillin, cephalexin, ceftriaxone), tetracyclines (tetracycline, doxycycline) and fluoroquinolones (ciprofloxacin, levofloxacin, marbofloxacin) groups of antimicrobials. Conversely, the highest resistance of the isolates was observed for amikacin (10/26=38%). Data on outcomes were available for 61% (16/26) of cats, of which 50% (8/16) died or were subjected to euthanasia due to severe complications (e.g., sepsis, pneumonia, and pleural effusion) secondary to disseminated/systemic infections, although no significant association was observed between Pasteurella species and the clinical-epidemiological findings studied. Our results contribute to the molecular identification of Pasteurella species and vigilance of multidrug-resistant bacteria that infect cats. Also, it highlights the need for the early diagnosis and therapy of feline pasteurellosis due to high mortality rates.
INDEX TERMS:
Feline pasteurellosis; Pasteurella multocida; MALDI-TOF MS; multidrug-resistant bacteria
RESUMO:
As espécies de Pasteurella são bactérias oportunistas que habitam a microbiota da cavidade oral e do trato respiratório superior de gatos, relacionadas a vários sinais clínicos em animais de companhia e humanos. A maioria dos estudos envolvendo pasteurelose felina têm sido descritos como relatos de casos, e a identificação de espécies baseada em métodos clássicos de classificação fenotípica. No entanto, número restrito de estudos têm focado grande número de gatos com pasteurelose, nos quais métodos moleculares tenham sido utilizados para o diagnóstico dos patógenos em nível da espécie. Neste cenário, foi investigada a identificação molecular de espécies de Pasteurella isoladas de 26 gatos domésticos com diferentes manifestações clínicas (e.g., abscessos cutâneos, pneumonia, conjuntivite, feridas, infecções do trato urinário, derrame pleural, piometra e infecção secundária à neoplasia), com base no diagnóstico por proteômica, utilizando a espectrometria de massas (Matrix-assisted laser desorption ionization time-of-flight mass spectrometry - MALDI-TOF MS). O perfil de sensibilidade microbiana in vitro dos isolados e dados epidemiológicos dos animais (com ênfase no desfecho dos casos) também foram avaliados. MALDI-TOF MS identificou predominantemente P. multocida (23/26=88,5%), seguido por P. dagmatis (2/26=7,7%) e P. canis (1/26=3,8%). Os isolados revelaram 100% de sensibilidade aos grupos de antimicrobianos beta-lactâmicos (amoxicilina/ácido clavulânico, ampicilina, cefalexina, ceftriaxona), tetraciclinas (tetraciclina, doxiciclina) e fluoroquinolonas (ciprofloxacino, levofloxacino, marbofloxacino). Por outro lado, a maior resistência dos isolados foi observada para a amicacina (10/26=38%). Os dados de desfecho estavam disponíveis para 61% (16/26) dos gatos, dos quais 50% (8/16) morreram ou foram submetidos à eutanásia devido a graves complicações secundárias a infecções disseminadas/sistêmicas, embora nenhuma associação estatística tenha sido observada entre as espécies de Pasteurella e os achados clínico-epidemiológicos estudados. Os resultados do presente estudo contribuem para a identificação molecular de espécies de Pasteurella e para a vigilância de bactérias multirresistentes que infectam gatos, indicando a necessidade de diagnóstico e tratamento precoces da pasteurelose felina devido às altas taxas de mortalidade.
TERMOS DE INDEXAÇÃO:
Pasteurelose felina; P. multocida; MALDI-TOF MS; bactérias multirresistentes
Introduction
It is roughly estimated that > 600 million cats live across the globe, of which over two-thirds (480 million) probably live as stray cats (Iapwa 2021). Domestic, free-ranging, or feral cats possess a habit of competing for food, for females during the reproductive period, or for the hierarchy of the environment, mainly among adult males (Stull 2022). It is common to experience skin infections secondary to bites and scratches, which can evolve into disseminated/systemic infections (De Cecco et al. 2021).
Pasteurella species are well-known bacteria that are opportunistic in nature, inhabit the oral cavity and upper respiratory tract of companions and livestock, and are related to a variety of clinical infections (Peng et al. 2019). It is estimated that the majority of cats can harbor Pasteurella species in the oral microbiota and may be potentially infectious for other animals (including cats). They are also the leading cause of pet-associated infections in humans, secondary to bites and scratches (Bula-Rudas & Olcott 2018, Stull 2022).
Cutaneous-subcutaneous abscesses represent the most common clinical sign related to feline pasteurellosis. However, systemic spread of the pathogen is common, causing fever, nasal secretion, weight loss, organ abscesses, ocular and urinary tract infections, pneumonia, peritonitis, pleural effusions, and encephalitis, usually with a poor prognosis (Lloret et al. 2013, Stull 2022).
The routine diagnosis of pasteurellosis in cats has been based on clinical-epidemiological findings, hematological and biochemical tests, imaging, bacteriological culture and phenotypic identification, and in vitro susceptibility tests (Stull 2022). Cephalosporins, fluoroquinolones, tetracyclines, and amoxicillin/clavulanic acid have been the main groups of antimicrobials used in the therapy of infections caused by Pasteurella species in cats (Lloret et al. 2013, Stull 2022).
In humans, Pasteurella species represent one of the most common bacteria causing cutaneous (local) and disseminated lesions secondary to cat bites and scratches (Mirzai et al. 2019, Piorunek et al. 2023). In addition, thin teeth of cats may develop deep perforating injuries in the tissues secondary to bites, with inoculation of saliva containing Pasteurella species, which probably has been underestimated due to the minimal appearance of the lesions (Kheiran et al. 2019).
Despite the severity of systemic infections by Pasteurella species in cats and guarded prognosis, most studies involving feline pasteurellosis have been described as case reports (Dolieslager et al. 2011, Lloret et al. 2013) and/or identification of species based on traditional bacteriological (phenotypic) tests (Wang et al. 2009, Awosile et al. 2018). Considering this scenario, we investigated the identification of Pasteurella at the species level using mass spectrometry and the in vitro susceptibility pattern of isolates recovered from a variety of clinical infections in 26 diseased domestic cats, as well as selected epidemiological findings, with emphasis on the outcome.
Materials and Methods
Ethics approval. This study was conducted under the Ethics Committee on Animal Use (CEUA) guidelines of the “Faculdade de Medicina Veterinária e Zootecnia” (School of Veterinary Medicine and Animal Science - FMVZ), “Universidade Estadual Paulista ‘Júlio de Mesquita Filho’” (Unesp), Botucatu/SP, Brazil (protocol number 0224/2021).
Animals and Pasteurella isolates. A convenient sampling of Pasteurella isolates obtained from diseased cats from the routine of a Teaching Veterinary Hospital and the “Laboratório de Diagnóstico Microbiológico” (Microbiological Diagnostic Laboratory) at FMVZ-Unesp, Botucatu/SP, Brazil, over 2019 and 2023, were used. The different clinical specimens were collected after rigorous antisepsis, kept refrigerated (4 to 8 °C), and immediately sent for microbiological culture in the aforementioned laboratory.
Bacteriological culture. A variety of clinical specimens (transtracheal wash, urine, organ fragments, and secretion from abscesses, ocular conjunctiva, pleural effusion, uterus, and wounds) were simultaneously cultured under aerobic conditions on bovine blood agar media (Oxoid™, São Paulo, Brazil) and selective MacConkey media (Oxoid™, São Paulo, Brazil), incubated at 37 °C, and kept for 72 hours. The isolated microorganisms were initially identified based on traditional morphotintorial and biochemical aspects (Quinn et al. 2011). Colonies compatible with Pasteurella species were subsequently submitted for identification at the species level using mass spectrometry. Clinical infections by Pasteurella species were considered > 10 colony-forming units (CFU) of isolates.
Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Isolates compatible with Pasteurella species in bacteriological culture were subjected to mass spectrometry for diagnostic confirmation. The MALDI-TOF MS technique (Bruker and Daltonics™, Bremen, Germany) was used, following the manufacturer’s recommendations. Three to four freshly isolated colonies (24 to 48 h) on blood agar were subjected to extraction with 20-40 µL of formic acid (70%) and centrifuged, aiming at bacterial lysis and release of proteins and formation of ions necessary for the formation of bacterial spectra. Approximately 15 minutes later, 20-40 µL of acetromil (100% P.A.) in the same proportion as formic acid (1:1) was added to each sample and centrifuged. Then, 1 µL of the solution for each sample was added to specific plates containing 96 wells (Bruker and Daltonics™, Bremen, Germany) and kept for approximately 20 minutes to dry at room temperature. The well with the dried samples was covered with 1 µL of matrix solution (2-cyano-4-hydroxycinnamic acid diluted with 50% acetonitrile and 2.5% trifluoroacetic acid). The plates were placed in the receptacle of the MALDI-TOF MS equipment (Bruker and Daltonics™, Bremen, Germany), operated with a 337-nm laser. Spectral data were analyzed between 2,000-20,000 m/z using FlexControl 3.3 software. Identifying microorganisms at the genus and species level was considered for isolates with spectra ≥ 1.7 and ≥ 2.0, respectively (Gonçalves et al. 2014).
In vitro susceptibility test. All isolates were subjected to in vitro susceptibility testing (disk diffusion method), according to the Clinical Laboratory Standards Institute (CLSI 2023, 2024) guidelines, using 11 antimicrobials belonging to five groups, predominantly indicated for the therapy of feline pasteurellosis, as follows: 1) aminoglycosides (amikacin, 30 μg), 2) beta-lactams and derivatives (amoxicillin/clavulanic acid, 30 μg; ampicillin, 10 μg; cephalexin, 30 μg; ceftriaxone, 30 μg), 3) fluoroquinolones (ciprofloxacin, 5 μg; levofloxacin 5 μg; marbofloxacin, 5 μg; norfloxacin, 10 μg), 4) macrolides (azithromycin, 15 μg), and 5) tetracyclines (doxycycline, 30 μg). Isolates resistant to ≥ 3 antimicrobials from different classes were considered multidrug-resistant ones (Magiorakos et al. 2012).
Epidemiological data. Selected epidemiological data, i.e., breed, gender, outcome (cure or death), and breeding conditions (restricted indoors, semi-domesticated, stray or feral animals), were assessed. The history of recent diagnosis (< 30 days) of immunosuppressive retrovirus (feline immunodeficiency virus - FIV and feline leukemia virus - FeLV) was investigated among studied animals. The periods (seasons) of the year (considered in Brazil) in which cases occurred were also assessed as follows: spring (September to November), summer (December to February), autumn (March to May) and winter (June to August). The age of the sampled animals was stratified as follows: < 1 year of age, 1 to < 5 years, > 5 to < 10 years, and > 10 years [20], considered young, young adults, adults, and elderly, respectively.
Statistical analysis. Associations between categorical variables (sex and age of animals, seasons, access to the street, and outcome), different clinical infections and identification of Pasteurella species were evaluated based on the Chi-square (or Fisher’s tests). The analyses were carried out using SAS OnDemand for Academics software (SAS Institute Inc., Cary/NC, USA), considering the level of significance results < 0.05.
Results
Selected epidemiological data
Of the 26 cats sampled, 92.3% (24/26) did not have a defined breed. Data on age were available from 88.5% (23/26) of the cats sampled. The ages ranged from two months to 16 years (average of 5.5 years), and among cats with known age, 21.7% (5/23) were < 1 year old (young), 21.7% (5/23) were between one and ≤ 5 years old (young adults), 26.1% (6/23) were > 5 and ≤ 10 years old (adults), and 30.5% (7/23) were > 10 years old (elderly). Among eight animals studied that died, 62.5% (5/8) were > 10 years old (considered elderly).
Among all cats, 65.4% (17/26) were males and 34.6% (9/26) were females. In addition, 53.8% (14/26) had access to the street, 30.8% (8/26) did not have access to the street, and 15.4% (4/26) could not obtain this data. Of the 26 cats, 34.6% (9/26) were sampled in spring, 26.9% (7/26) in summer, 11.6% (3/26) in autumn, and 26.9% (7/26) in winter. Among the 26 cats studied, only 15.4% (4/26) had been recently tested (< 30 days) for FIV and FeLV, of which two males were reactive and, therefore, this information was not considered for further analyses.
Outcome
Data on outcomes were available in 61% (16/26) of animals. Of these animals, 50% (8/16) died or were subjected to euthanasia due to severe complications secondary to disseminated forms, including sepsis, pneumonia, and pleural effusion. In turn, among the remaining 50% (8/16) of animals with known outcomes, 75% (6/8 = 75%) recovered after treatment, mainly from skin lesions (or local infections).
Microbiological findings and mass spectrometry identification of Pasteurella species
Among the 26 cats sampled, rounded, mucoid, grayish, nonhemolytic colonies measuring ~1 mm in diameter were isolated between 24 and 48 hours in blood agar media (Fig.1), phenotypically compatible with Pasteurella species. All these isolates were subjected to Gram staining, revealing Gram-negative rods to coccobacillary bacteria with bipolar aspect tendency.
Pasteurella multocida isolated from a cat with pneumonia identified at the species level by mass spectrometry. Note rounded, mucoid, grayish, nonhemolytic colonies measuring ~1 mm in diameter isolated in blood agar media, after 48 hours of incubation, under aerobic conditions at 37 °C.
The isolates suggestive of Pasteurella species, subjected to mass spectrometry diagnosis, revealed a predominance of P. multocida (23/26 = 88.5%) identification, followed in a minor frequency by P. dagmatis (2/26 = 7.7%) and P. canis (1/26 = 3.8%) (Table 1).
In vitro antimicrobial susceptibility pattern
The isolates revealed 100% susceptibility to beta-lactam (amoxicillin/clavulanic acid, ampicillin, cephalexin, ceftriaxone), fluoroquinolones (ciprofloxacin, levofloxacin, marbofloxacin), and tetracyclines (tetracycline, doxycycline) groups of antimicrobials. In turn, the highest resistance of the isolates was observed for amikacin (10/26=38%) (Table 2). No multidrug-resistant isolates were identified.
Clinical signs
Abscesses (11/26 = 42.3%), pneumonia (3/26 = 11.5%), conjunctivitis (3/26 = 11.5%), open wounds (3/26 = 11.5%), urinary tract infections (2/26 = 7.7%), pleural effusion (2/26 = 7.7%), pyometra (1/26 = 3.9%), and infection secondary to neoplasia (1/26 = 3.9%) were the clinical signs observed among the 26 cats with identification of Pasteurella species (Table 1).
Statistical analysis
No significant association (p> 0.05) was observed between Pasteurella species and the clinical-epidemiological findings studied.
Discussion
The population of domestic, free-ranging and feral cats has increased around the world (Iapwa 2021), including in Brazil, which possesses one of the greatest number of cats worldwide (Moutinho et al. 2019). Despite the psychosocial and well-being benefits to owners with the presence of domestic cats in households, their close contact with felines offers a risk of the transmission for humans of pet-associated diseases that have a zoonotic nature (Esch & Petersen 2013), a fact that deserves concern in the One Health concept (Kheiran et al. 2019, Mirzai et al. 2019), and may be considered a motivation of the current study.
Pasteurella constitute a group of bacteria that inhabit the oral microbiota and upper respiratory tract of domestic animals, whose infections in companion animals are strongly associated with the opportunistic behavior of the pathogen (Stull 2022). More than 15 Pasteurella species are well-known, although P. multocida, P. dagmatis and P. stomatis have been frequently isolated from a set of clinical infections in domestic animals (Quinn et al. 2011), particularly cats (Stull 2022). Among the 26 cats sampled, 88.5% of isolates were identified as P. multocida based on mass spectrometry diagnosis, followed in minor frequency by P. dagmatis and P. canis, reinforcing the predominance of P. multocida as a primary species in clinical infections in domestic cats (Lloret et al. 2013, Stull 2022).
Different epidemiological factors must be considered in Pasteurella infections in domestic cats, including sex, age and breed of the animals, territorial habits of the felines (disputes over dominance in the environment, food and females in estrus), underlying conditions (coinfection with FIV and FeLV), and the access to the external environment of the household (Love et al. 2000, Walker et al. 2000, Wang et al. 2009). Nonetheless, no significant associations were observed between these selected epidemiological variables, clinical signs, and proteomic identification of Pasteurella species.
Despite no statistical association of epidemiological data studied, the high prevalence of mixed-breed cats could be credited to an increase adoption of stray cats in Brazil (Moutinho et al. 2019) or to the low socioeconomic profile of the owners referred to the routine of the Teaching Veterinary Hospital studied.
A wide variation in the age of the cats sampled was observed (2 months to 16 years old), indicating that feline pasteurellosis can occur in any age group. Nonetheless, a highly lethal outcome was observed among cats studied considered elderly (> 10 years old), which could be attributed to the development of clinical complications (i.e., sepsis, pneumonia, and pleural effusion) secondary to systemic spread of the pathogen in debilitated animals (Giordano et al. 2015, Stull 2022).
Among all cats studied, 65.4% (17/26) were males, and 53.8% (14/26) had access to the street. In addition, 34.6% (9/26) of Pasteurella infections occurred in spring and 26.9% in summer, considered hot periods for reproductive action. The free access of males to the street agrees with similar studies involving feline pasteurellosis (Freshwater 2008, Lloret et al. 2013), which could facilitate respiratory infections through direct contact between cats or result in cutaneous lesions secondary to bites or scratches due to the territorial habits of male cats (i.e., disputes over food, females in estrus and/or dominance of the territory). The territorial behavior of cats could favor the traumatic inoculation of Pasteurella species in skin tissue and opportunistic infections of the pathogen (Johnston et al. 2001, Wang et al. 2009, Lloret et al. 2013, Giordano et al. 2015).
Traumatic inoculation in the skin of the pathogen by bites or scratches and inhalation of the agent represent frequent routes of feline infections by Pasteurella species. In addition, cat licking is also considered an infection route for Pasteurella species due to the grooming of felines, enabling contamination of previous lesions by owner saliva (Stull 2022). In these routes of infections, the animals commonly evolve to cutaneous abscesses, cellulitis or disseminated infections and, occasionally, sepsis and development of abscesses in organs (Quinn et al. 2011, De Cecco et al. 2021, Wei et al. 2021, Stull 2022). Likewise, in the current study, abscesses (42.3%) represented the most common clinical sign among 26 diseased cats, followed by a set of other clinical manifestations in minor frequency, i.e., pneumonia, conjunctivitis, open wounds, urinary tract infections, pleural effusion, pyometra, and infection secondary to neoplasia. The predominance of cutaneous-subcutaneous lesions in cats studied agrees with similar studies, in which skin lesions have been considered the most common clinical sign of feline pasteurellosis, possibly due to traumatic inoculation of the agent secondary to bites and scratches (Lloret et al. 2013, Giordano et al. 2015, Stull 2022). In addition, the variety of clinical manifestations observed in 26 diseased cats studied reinforces the opportunistic behavior of Pasteurella-induced infections in domestic cats (Quinn et al. 2011, Stull 2022).
Except for amikacin, azithromycin and norfloxacin, all other antimicrobials showed > 95% in vitro efficacy against Pasteurella isolates among cats sampled. Conversely, the highest resistance of the isolates was observed to amikacin (10/26 = 38%), followed by azithromycin and norfloxacin, whereas no multidrug-resistant Pasteurella isolates were reported.
Beta-lactams and derivatives (amoxicillin/clavulanic acid, ampicillin, cephalexin, ceftriaxone) and tetracyclines (doxycycline and tetracycline) groups/antimicrobials showed 100% in vitro efficacy against the three Pasteurella species isolated in the current study. Furthermore, ciprofloxacin, levofloxacin and marbofloxacin (fluoroquinolone group) also showed 100% efficacy against the isolates, except for norfloxacin, which revealed one resistant P. multocida isolates. These results are consistent with similar in vitro studies of the susceptibility patterns of Pasteurella species isolated from cats (Freshwater 2008, De Cecco et al. 2021) and P. multocida isolated from humans bitten by cats in the USA (Westling et al. 2006). These findings indicate that, in general, antimicrobials belonging to the groups of beta-lactams, tetracyclines and fluoroquinolones represent good therapeutic options for the treatment of feline pasteurellosis, probably due to the broad spectrum of action (including Gram-negative bacteria), and the high therapeutic concentrations reached into cells and tissues (Lloret et al. 2013, Stull 2022).
Some groups of antimicrobials have shown effectiveness in treating Pasteurella infections in domestic cats. Nonetheless, it is recommended that, if possible, treatments be carried out with previous support of in vitro susceptibility tests of the isolates, which can increase the success of treatment (Guiguère et al. 2013), as well as avoid the nonrational or overusing of antimicrobials, which is related to increase of selective pressure for multidrug-resistant bacteria (Magiorakos et al. 2012), a global emerging concern (Wei et al. 2021).
Despite the clinical severity and poor prognosis of systemic disorders-related Pasteurella infections in domestic felines (Giordano et al. 2015), most studies involving feline pasteurellosis have been described as case reports (Dolieslager et al. 2011, De Cecco et al. 2021), and the diagnosis based on traditional phenotypic tests (Foster et al. 2004, Freshwater 2008, Wang et al. 2009, Giordano et al. 2015, Awosile et al. 2018). Conversely, in the current study, Pasteurella isolates obtained from 26 diseased domestic cats were diagnosed at the species level based on proteomics (MALDI-TOF MS), which has been revealed to be a reliable and fast technique with high discriminatory power for the identification of bacterial and yeast species, enabling etio-epidemiological studies with microorganisms from animal and human origin (Kuhnert et al. 2012, Zangenah et al. 2013).
Data on outcomes were available in 61% (16/26) of animals. Of these, 50% (8/16) died or were subjected to euthanasia due to severe complications, such as sepsis, pneumonia, and pleural effusion. This finding highlights the poor prognosis in cases of dissemination or systemic infections by Pasteurella species (Giordano et al. 2015, Stull 2022) and the need for early diagnosis and therapy approaches of feline pasteurellosis due to high mortality rates of disease.
Clinical pasteurellosis in cats has been related to debilitated animals or those coinfected with an immunosuppressive retrovirus (Stull 2022). Nonetheless, among 26 cats sampled, only two (7.7%) males showed reactions to FIV and FeLV tests, limiting the assessment to the impact of coinfection of Pasteurella species and these immunosuppressive diseases in cats studied.
Pasteurella species represent a predominant group of bacteria isolated from lesions in humans secondary to being bitten and scratched by cats (Mirzai et al. 2019, Lloret et al. 2013, Mu et al. 2020) due to inhabiting oral microbiota of normal cats (Portilho et al. 2024). In addition, Pasteurella isolates recovered from owners and cats that inhabit the same households have revealed similarities regarding the virulence and in vitro antimicrobial susceptibility profile, indicating that cats may be considered potential reservoirs of the pathogen from other animals and humans (Ujvári et al. 2019). Also, domestic cats possess a thin thickness of teeth that favor deep lesions in skin inoculated by their saliva, which may induce cutaneous-subcutaneous lesions and systemic dissemination of the pathogen (Love et al. 2000, Kheiran et al. 2019), which deserve relevance in human health.
Convenience sampling, a lack of identification of Pasteurella serogroups/subspecies, and no investigation of comorbidity conditions or immunosuppressive viral diseases (i.e., FIV and FeLV) in a total of animals studied may be considered limiting factors of the current study.
Conclusion
Overall, the identification of Pasteurella species in 26 domestic cats with different clinical infections was investigated using proteomics (mass spectrometry), which revealed a predominance of P. multocida species and a high occurrence of skin lesions, mainly in male cats, in addition to a high fatal evolution of systemic or disseminated infections. Our results contribute to the molecular identification of Pasteurella species, the vigilance of multidrug-resistant isolates, and concerns regarding high mortality rates of disseminated/systemic feline pasteurellosis.
Acknowledgments
We thank the “Conselho Nacional de Desenvolvimento Científico e Tecnológico” (CNPq), Brazil, for the research productivity fellowship (PQ) given to Márcio Garcia Ribeiro (#310345/2020-0).
References
- Awosile BB, McClure JT, Saab ME, Heider LC. Antimicrobial resistance in bacteria isolated from cats and dogs from the Atlantic Province, Canada from 1994-2013. Can Vet J 2018;59(8):885-893. PMid:30104781
-
Bula-Rudas FJ, Olcott JL. Human and animal bites. Pediatr Rev 2018; https://doi.org/10.1542/pir.2017-0212, PMid:30275032
» https://doi.org/10.1542/pir.2017-0212 - CLSI. Performance standards for antimicrobial disk and dilution susceptibility test for bacteria isolated from animals (CLSI VET 01S). 7th ed. Wayne: Clinical and Laboratory Standards Institute; 2024.
- CLSI. Performance standards of antimicrobial susceptibility testing (M100). 33th ed. Wayne: Clinical and Laboratory Standards Institute; 2023.
-
De Cecco BS, Carossino M, Del Piero F, Wakamatsu N, Mitchell MS, Fowlkes NW, Langohr IM. Meningoencephalomyelitis in domestic cats: 3 cases of Pasteurella multocida infection and literature review. J Vet Diagn Invest 2021; https://doi.org/10.1177/10406387211034484, PMid:34301172
» https://doi.org/10.1177/10406387211034484 -
Dolieslager SMJ, Riggio MP, Lennon A, Lappin DF, Johnston N, Taylor D, Bennett D. Identification of bacteria associated with feline chronic gingivostomatitis using culture-dependent and culture-independent methods. Vet Microbiol 2011; https://doi.org/10.1016/j.vetmic.2010.08.002, PMid:20828946
» https://doi.org/10.1016/j.vetmic.2010.08.002 -
Esch KJ, Petersen CA. Transmission and epidemiology of zoonotic protozoal diseases of companion animals. Clin Microbiol Rev 2013; https://doi.org/10.1128/CMR.00067-12, PMid:23297259
» https://doi.org/10.1128/CMR.00067-12 -
Foster SF, Martin P, Allan GS, Barrs VR, Malik R. Lower respiratory tract infections in cats: 21 cases (1995-2000). J Feline Med Surg 2004; https://doi.org/10.1016/j.jfms.2003.11.006, PMid:15135354
» https://doi.org/10.1016/j.jfms.2003.11.006 -
Freshwater A. Why your housecat’s trite little bite could cause you quite a fright: a study of domestic felines on the occurrence and antibiotic susceptibility of Pasteurella multocida Zoonoses Public Health 2008; https://doi.org/10.1111/j.1863-2378.2008.01152.x, PMid:18811910
» https://doi.org/10.1111/j.1863-2378.2008.01152.x -
Giordano A, Dincman T, Clyburn BE, Steed LL, Rockey DC. Clinical features and outcomes of Pasteurella multocida infection. Medicine, United States 2015; https://doi.org/10.1097/MD.0000000000001285, PMid:26356688
» https://doi.org/10.1097/MD.0000000000001285 -
Gonçalves JL, Tomazi T, Barreiro JR, Braga PAC, Ferreira CR, Araújo Junior JP, Eberlin MN, Santos MV. Identification of Corynebacterium spp. isolated from bovine intramammary infections by matrix-assisted laser desorption ionization-time of flight mass spectrometry. Vet Microbiol 2014; https://doi.org/10.1016/j.vetmic.2014.06.028, PMid:25086477
» https://doi.org/10.1016/j.vetmic.2014.06.028 -
Guiguère S, Prescoott JF, Dowling PM. Antimicrobial therapy in veterinary medicine. 2013; https://doi.org/10.1002/9781118675014
» https://doi.org/10.1002/9781118675014 -
IAPWA. Neuter strays for healthier happier lives. International Aid for the Protection & Welfare of Animals; 2021. Accessed September 3, 2024. https://iapwa.org/neuter-strays-for-healthier-happier-lives/
» https://iapwa.org/neuter-strays-for-healthier-happier-lives/ - Johnston SD, Kustriz MVR, Olson PNS. Canine and feline theriogenology. Philadelphia: Saunders; 2001.
-
Kheiran A, Palial V, Rollett R, Wildin CJ, Chatterji U, Singh HP. Cat bite: an injury not to underestimate. J Plastic Surg Hand Surg 2019; https://doi.org/10.1080/2000656X.2019.1637750, PMid:31287352
» https://doi.org/10.1080/2000656X.2019.1637750 -
Kuhnert P, Bisgaard M, Korczak BM, Schwendener S, Christensen H, Frey J. Identification of animal Pasteurellaceae by MALDI-TOF mass spectrometry. J Microbiol Methods 2012; https://doi.org/10.1016/j.mimet.2012.02.001, PMid:22343217
» https://doi.org/10.1016/j.mimet.2012.02.001 -
Lloret A, Egberink H, Addie D, Belák S, Boucraut-Baralon C, Frymus T, Gruffydd-Jones T, Hartmann K, Hosie MJ, Lutz H, Marsilio F, Möstl K, Pennisi MG, Radford AD, Thiry E, Truyen U, Horzinek MC. Pasteurella multocida infection in cats: ABCD guidelines on prevention and management. J Feline Med Surg 2013; https://doi.org/10.1177/1098612X13489215, PMid:23813817
» https://doi.org/10.1177/1098612X13489215 -
Love DN, Malik R, Norris JM. Bacteriological warfare amongst cats: what have we learned about cat bite infections? Vet Microbiol 2000; https://doi.org/10.1016/S0378-1135(00)00186-3, PMid:10808087
» https://doi.org/10.1016/S0378-1135(00)00186-3 -
Magiorakos A-P, Srinivasan A, Carey RB, Carmeli Y, Falagas ME, Giske CG, Harbarth S, Hindler JF, Kahlmeter G, Olsson-Liljequist B, Paterson DL, Rice LB, Stelling J, Struelens MJ, Vatopoulos A, Weber JT, Monnet DL. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect 2012; https://doi.org/10.1111/j.1469-0691.2011.03570.x, PMid:21793988
» https://doi.org/10.1111/j.1469-0691.2011.03570.x -
Mirzai S, Rifai AO, Tidrick A, Huang Q, Hale J. A case report on Pasteurella multocida peritoneal dialysis-associated peritonitis: when cats think medical equipment are toys. Case Reports Nephrol 2019; https://doi.org/10.1155/2019/5150695, PMid:31934471
» https://doi.org/10.1155/2019/5150695 -
Moutinho FFB, Serra CMB, Valente LCM. Situação pós-adoção dos animais adotados junto a uma ONG de proteção animal no Estado do Rio de Janeiro. Ciênc Anim Bras 2019; https://doi.org/10.1590/1809-6891v20e-43777
» https://doi.org/10.1590/1809-6891v20e-43777 -
Mu H, Yang M, Zhang Y, Zhang Y, Wang J, Yuan W, Rong S. Pet-related Pasteurella multocida induced peritonitis in peritoneal dialysis: a case report and review of the literatures. BMC Nephrol 2020; https://doi.org/10.1186/s12882-020-01765-1, PMid:32192435
» https://doi.org/10.1186/s12882-020-01765-1 -
Peng Z, Wang X, Zhou R, Chen H, Wilson BA, Wu B. Pasteurella multocida: genotypes and genomics. Microbiol Mol Biol Rev 2019; https://doi.org/10.1128/MMBR.00014-19, PMid:31484691
» https://doi.org/10.1128/MMBR.00014-19 -
Piorunek M, Brajer-Luftmann B, Walkowiak J. Pasteurella multocida infection in humans. Pathogens 2023; https://doi.org/10.3390/pathogens12101210, PMid:37887726
» https://doi.org/10.3390/pathogens12101210 -
Portilho FVR, Nóbrega J, Almeida BO, Bello TS, Paz PJL., Oliveira AL, Bosco SMG, Rall VLM, Giuffrida R, Ribeiro MG. The polymicrobial nature of the oral cavity and claws of cats diagnosed by mass spectrometry and next-generation sequencing. Microb Pathog 2024; https://doi.org/10.1016/j.micpath.2024.106765, PMid:38944215
» https://doi.org/10.1016/j.micpath.2024.106765 - Quinn PJ, Markey BK, Leonard FC, Fitzpatrick ES, Fanning S, Harting PJ. Pasteurella species, Mannheimua haemolytica and Bibersteinia trehalosi, p.300-308. In: Ibid. Veterinary Microbiology and Microbial Disease. 2nd ed. Chichester: Willey-Blackwell; 2011.
- Stull JW. Companion animal in immunocompromised and other high-risk human populations: pasteurellosis, p.228-229. In: Sykes JE. Greene’s infectious diseases of the dog and cat. 5th ed. Elsevier Health Sciences; 2022.
-
Ujvári B, Weiczner R, Deim Z, Terhes G, Urbán E, Tóth AR, Magyar T. Characterization of Pasteurella multocida strains isolated from human infections. Comp Immunol Microbiol Infect Dis 2019; https://doi.org/10.1016/j.cimid.2018.12.008, PMid:30961816
» https://doi.org/10.1016/j.cimid.2018.12.008 -
Walker AL, Jang SS, Hirsh DC. Bacteria associated with pyothorax of dogs and cats: 98 cases (1989-1998). J Am Vet Med Assoc 2000; https://doi.org/10.2460/javma.2000.216.359, PMid:10668533
» https://doi.org/10.2460/javma.2000.216.359 -
Wang AL, Ledbetter EC, Kern TJ. Orbital abscess bacterial isolates and in vitro antimicrobial susceptibility patterns in dogs and cats. Vet Ophthalmol 2009; https://doi.org/10.1111/j.1463-5224.2008.00687.x, PMid:19261163
» https://doi.org/10.1111/j.1463-5224.2008.00687.x -
Wei A, Dhaduk N, Taha B. Wrist abscess due to drug-resistant Pasteurella multocida IDCases 2021; https://doi.org/10.1016/j.idcr.2021.e01277, PMid:34522615
» https://doi.org/10.1016/j.idcr.2021.e01277 -
Westling K, Farra A, Cars B, Ekblom AG, Sandstedt K, Settergren B, Wretlind B, Jorup C. Cat bite wound infections: a prospective clinical and microbiological study at three emergency wards in Stockholm, Sweden. J Infect 2006; https://doi.org/10.1016/j.jinf.2006.01.001, PMid:16483663
» https://doi.org/10.1016/j.jinf.2006.01.001 -
Zangenah S, Güleryüz G, Boräng S, Ullberg M, Bergman P, Özenci V. Identification of clinical Pasteurella isolates by MALDI-TOF - a comparison with VITEK 2 and conventional microbiological methods. Diagn Microbiol Infect Dis 2013; https://doi.org/10.1016/j.diagmicrobio.2013.06.024, PMid:23886788
» https://doi.org/10.1016/j.diagmicrobio.2013.06.024
-
Data Availability Statement
Data availability statement – The corresponding author is responsible for the data’s maintenance, storage and accountability. All data were preserved on personal computers and in the cloud. The study data were published in the article at “Pesquisa Veterinária Brasileira” journal and are contained in the first author’s dissertation (Carolina Aparecida Rodrigues) and the “Universidade Estadual Paulista ‘Júlio de Mesquita Filho’” (Unesp) repository.
Data availability statement – The corresponding author is responsible for the data’s maintenance, storage and accountability. All data were preserved on personal computers and in the cloud. The study data were published in the article at “Pesquisa Veterinária Brasileira” journal and are contained in the first author’s dissertation (Carolina Aparecida Rodrigues) and the “Universidade Estadual Paulista ‘Júlio de Mesquita Filho’” (Unesp) repository.
