Klebsiella-induced infections in domestic species: a case-series study in 697 animals (1997-2019).

Klebsiella species, particularly K. pneumoniae, are well-known opportunistic enterobacteria related to complexity of clinical infections in humans and animals, commonly refractory to conventional therapy. The domestic animals may represent a source of the pathogenic and multidrug-resistant Klebsiella species to humans. Nevertheless, most studies involving Klebsiella-induced infections in domestic animals are restricted to case reports or outbreaks. We retrospectively investigated selected epidemiological data, clinical aspects, and in vitro susceptibility pattern of 697 non-repetitive Klebsiella infections in livestock and companion species (1997-2019). The isolates were obtained from different clinical disorders from dogs (n = 393), cattle (n = 149), horses (n = 98), cats (n = 27), pigs (n = 22), sheep (n = 5), goats (n = 2), and buffalo (n = 1), except four isolates from subclinical bovine mastitis. Urinary (223/697 = 32%), enteric (117/697 = 16.8%), mammary (85/697 = 12.2%), reproductive (85/697 = 12.2%), and respiratory disorders (67/697 = 9.6%) were the most common clinical manifestations. Other miscellaneous clinical pictures (116/697 = 16.6%) included abscesses, otitis, hepatitis, conjunctivitis, pyodermitis, sepsis, and encephalitis. Norfloxacin (183/245 = 74.7%) and gentamicin (226/330 = 68.5%) were the most effective antimicrobials. High in vitro resistance of the isolates was seen to ampicillin (326/355 = 91.8%), amoxicillin/clavulanic acid (25/62 = 40.3%), and trimethoprim/sulfamethoxazole (100/252 = 39.7), and multidrug resistance to ≥ 3 classes of antimicrobials was found in 20.4% (142/697) isolates. Wide variety of clinical manifestations of Klebsiella-induced infections was observed, with a predominance of urinary, enteric, mammary, reproductive, and respiratory tract disorders, reinforcing opportunistic behavior of agent. Poor in vitro efficacy was observed to some conventional antimicrobials and ~ 20% of isolates exhibited resistance pattern, reinforcing the need for proper use of drugs on therapy approaches in domestic animals to avoid multidrug-resistant bacteria, an emergent global concern.

Identification of Mycobacterium species and Rhodococcus equi in peccary lymph nodes.

Mycobacterium species and the virulence-associated proteins (vapA, vapB, and vapN genes) of Rhodococcus equi isolated from 330 lymph nodes of collared peccaries (Tayassu tajacu) and white-lipped peccaries (Tayassu pecari) intended for human consumption were investigated. Thirty-six (10.9%) R. equi strains were isolated; 3.3% (n = 11/330) were from white-lipped peccary lymph nodes, and 7.6% (25/330) were from collared peccary lymph nodes. Among the 11 isolates of R. equi from the white-lipped peccaries, 90.9% (n = 10/11) were obtained from the mesenteric lymph nodes, and only 9.1% (n = 1/10) were obtained from the mediastinal lymph nodes. In the 25 isolates of R. equi obtained from the collared peccaries, 40.0% (n = 10/25) were recovered from the mesenteric lymph nodes, 36% (n = 9/25) from the submandibular lymph nodes, and 24.0% (n = 6/25) from the mediastinal lymph nodes. No vapA, vapB, or vapN genes (plasmidless) or three host-associated types (pVAPA, pVAPB, and pVAPN) were identified among the R. equi isolates. Mycobacterium species were isolated in 3.03% (n = 10/330) of all the lymph nodes analyzed. Among the 10 mycobacterial isolates, 60% (n = 6/10) were from the white-lipped peccary lymph nodes, and 40% (n = 4/10) were from the collared peccary lymph nodes. Ten Mycobacterium species were detected by PCR-PRA with a predominance of M. avium type 1. Sequencing of the hsp65 and rpob genes revealed mycobacteria that were saprophytic (M. sinense and M. kumamotonense) and potentially pathogenic (M. colombiense and M. intracellulare) to humans and animals. To our knowledge, this is the first description of R. equi and/or mycobacterial species identified in the lymph nodes of peccary specimens. R. equi (plasmidless) and the mycobacterial species described here have been reported as causes of pulmonary and extrapulmonary infections in both immunocompetent and immunocompromised humans.

Nontuberculous mycobacteria in milk from positive cows in the intradermal comparative cervical tuberculin test: implications for human tuberculosis infections.

Although the tuberculin test represents the main in vivo diagnostic method used in the control and eradication of bovine tuberculosis, few studies have focused on the identification of mycobacteria in the milk from cows positive to the tuberculin test. The aim of this study was to identify Mycobacterium species in milk samples from cows positive to the comparative intradermal test. Milk samples from 142 cows positive to the comparative intradermal test carried out in 4,766 animals were aseptically collected, cultivated on Lowenstein-Jensen and Stonebrink media and incubated for up to 90 days. Colonies compatible with mycobacteria were stained by Ziehl-Neelsen to detect acid-fast bacilli, while to confirm the Mycobacterium genus, conventional PCR was performed. Fourteen mycobacterial strains were isolated from 12 cows (8.4%). The hsp65 gene sequencing identified M. engbaekii (n=5), M. arupense (n=4), M. nonchromogenicum (n=3), and M. heraklionense (n=2) species belong to the Mycobacterium terrae complex. Despite the absence of M. tuberculosis complex species in the milk samples, identification of these mycobacteria highlights the risk of pathogen transmission from bovines to humans throughout milk or dairy products, since many of mycobacterial species described here have been reported in pulmonary and extrapulmonary diseases both in immunocompetent and immunocompromised people.

Diagnosis of mycobacteria in bovine milk: an overview.

Tuberculosis remains as the world's biggest threat. In 2014, human tuberculosis ranked as a major infectious disease by the first time, overcoming HIV death rates. Bovine tuberculosis is a chronic disease of global distribution that affects animals and can be transmitted to humans by the consumption of raw milk, representing a serious public health concern. Despite the efforts of different countries to control and eradicate bovine tuberculosis, the high negative economic impact on meat and milk production chains remains, given the decreased production efficiency (approximately 25%), the high number of condemned carcasses, and increased animal culling rates. This scenario has motivated the establishment of official programs based on regulations and diagnostic procedures. Although Mycobacterium tuberculosis and Mycobacterium bovis are the major pathogenic species to humans and bovines, respectively, nontuberculous mycobacteria within the Mycobacterium genus have become increasingly important in recent decades due to human infections, including the ones that occur in immunocompetent people. Diagnosis of mycobacteria can be performed by microbiological culture from tissue samples (lymph nodes, lungs) and secretions (sputum, milk). In general, these pathogens demand special nutrient requirements for isolation/growth, and the use of selective and rich culture media. Indeed, within these genera, mycobacteria are classified as either fast- or slow-growth microorganisms. Regarding the latter ones, incubation times can vary from 45 to 90 days. Although microbiological culture is still considered the gold standard method for diagnosis, molecular approaches have been increasingly used. We describe here an overview of the diagnosis of Mycobacterium species in bovine milk.

Multidrug resistant bacteria isolated from septic arthritis in horses / Bactérias multirresistentes isoladas de artrite séptica equina

Septic arthritis is a debilitating joint infectious disease of equines that requires early diagnosis and immediate therapeutic intervention to prevent degenerative effects on the articular cartilage, as well as loss of athletic ability and work performance of the animals. Few studies have investigated the etiological complexity of this disease, as well as multidrug resistance of isolates. In this study, 60 horses with arthritis had synovial fluid samples aseptically collected, and tested by microbiological culture and in vitro susceptibility test (disk diffusion) using nine antimicrobials belonging to six different pharmacological groups. Bacteria were isolated in 45 (75.0%) samples, as follows: Streptococcus equi subsp. equi (11=18.3%), Escherichia coli (9=15.0%), Staphylococcus aureus (6=10.0%), Streptococcus equi subsp. zooepidemicus (5=8.3%), Staphylococcus intermedius (2=3.3%), Proteus vulgaris (2=3.3%), Trueperella pyogenes (2=3.3%), Pseudomonas aeruginosa (2=3.3%), Klebsiella pneumoniae (1=1.7%), Rhodococcus equi (1=1.7%), Staphylococcus epidermidis (1=1.7%), Klebsiella oxytoca (1=1.7%), Nocardia asteroides (1=1.7%), and Enterobacter cloacae (1=1.7%). Ceftiofur was the most effective drug (>70% efficacy) against the pathogens in the disk diffusion test. In contrast, high resistance rate (>70% resistance) was observed to penicillin (42.2%), enrofloxacin (33.3%), and amikacin (31.2%). Eleven (24.4%) isolates were resistant to three or more different pharmacological groups and were considered multidrug resistant strains. The present study emphasizes the etiological complexity of equine septic arthritis, and highlights the need to institute treatment based on the in vitro susceptibility pattern, due to the multidrug resistance of isolates. According to the available literature, this is the first report in Brazil on the investigation of the etiology. of the septic arthritis in a great number of horses associated with multidrug resistance of the isolates.(AU)

Artrite séptica é uma artropatia infecciosa debilitante de equinos, que requer diagnóstico precoce e intervenção terapêutica imediata, com intuito de evitar a degeneração de a cartilagem articular e a perda da capacidade atlética e de trabalho dos animais. Poucos estudos têm investigado a complexidade etiológica da afecção, bem como a presença de multirresistência dos isolados aos antimicrobianos. Foram investigados 60 equinos portadores de artrite, submetidos à colheita asséptica de líquido sinovial para a realização de cultivo microbiológico e teste de sensibilidade microbiana in vitro (difusão com discos) com nove antimicrobianos pertencentes a seis diferentes grupos farmacológicos. Foi obtido isolamento microbiano em 45 (75,0%) amostras, como segue: Streptococcus equi subsp. equi (11=18,3%), Escherichia coli (9=15,0%), Staphylococcus aureus (6=10,0%), Streptococcus zooepidemicus (5=8,3%), Staphylococcus intermedius (2=3,3%), Proteus vulgaris (2=3,3%), Trueperella pyogenes (2=3,3%), Pseudomonas aeruginosa (2=3,3%), Klebsiella pneumoniae (1=1,7%), Rhodococcus equi (1=1,7%), Staphylococcus epidermidis (1=1,7%), Klebsiella oxytoca (1=1,7%), Nocardia asteroides (1=1,7%) e Enterobacter cloacae (1=1,7%). Ceftiofur foi o antimicrobiano mais efetivo (>70% eficácia) in vitro diante dos patógenos. Em contraste, alta resistência dos isolados (>70% de resistência) foi observada para penicilina (42,2%), enrofloxacino (33,3%) e amicacina (31,2%). Onze (24,4%) isolados foram resistentes a três ou mais diferentes grupamentos de fármacos e considerados com resistência múltipla aos antimicrobianos. O presente estudo enaltece a complexidade etiológica envolvida na artrite séptica em equinos e ressalta a necessidade de instituir o tratamento dos animais com respaldo de testes de sensibilidade microbiana in vitro em virtude da resistência múltipla dos isolados. De acordo com a literatura consultada, esta é a primeira descrição no país da etiologia da artrite séptica em grande número de equinos associada a multirresistência dos isolados aos fármacos testados.(AU)

Successful therapy in unusual generalized Dermatophilus congolensis infection in a calf based on modified in vitro disk diffusion test / Sucesso no tratamento de infecção generalizada em bezerra por Dermatophilus congolensis baseado em teste in vitro modificado de difusão com discos

Bovine dermatophilosis is a dermatitis characterized by typical focal or localized lesions with "paintbrush" aspect and occasionally as disseminated cutaneous disease. We report the case of a one-year-old Nelore female with history of chronic cutaneous disseminated lesions that appeared immediately after a rainfall period. Serous to purulent exudates, hair with tufted appearance, hyperkeratotic, non-pruritic, hardened, yellowish to brown, and coalescent crusty lesions were observed distributed all over its body. Removal of the crusts revealed ulcerated or hemorrhagic areas, with irregular elevated crusts like "paintbrush". Microbiological diagnosis enabled the identification of a microorganism, the Dermatophilus congolensis. Despite disseminated and chronic lesions, we obtained a successful therapy with parenteral therapy using long-acting tetracycline based on modified in vitro disk diffusion test. The present report highlights success therapy in uncommon generalized bovine dermatophilosis with selection of first-choice drugs based on modified in vitro susceptibility test, and need of responsible use of antimicrobials in livestock.(AU)

A dermatofilose bovina é uma dermatite caracterizada por lesões focais ou localizadas com aspecto de "pincel" e, ocasionalmente, como lesão cutânea disseminada. Relata-se o caso de uma fêmea bovina de um ano de idade, que foi atendida apresentando história de lesões cutâneas crônicas imediatamente após um período de alta pluviosidade. Ao exame clínico, lesões serosas a purulentas, com hiperqueratose, coalescentes, não pruriginosas, ressecadas, de coloração amarelada à acinzentada foram observadas distribuídas de modo generalizado pelo animal. A remoção das crostas revelou áreas ulceradas ou hemorrágicas, com crostas irregulares e elevadas semelhantes a "pincel". O diagnóstico microbiológico possibilitou a identificação do micro-organismo Dermatophilus congolensis. Apesar das lesões disseminadas e crônicas, a cura do animal foi obtida com tratamento parenteral usando oxitetraciclina de longa duração, baseado em teste in vitro de sensibilidade microbiana modificado. O presente relato ressalta o sucesso no tratamento de caso incomum de lesões generalizadas de dermatofilose bovina com respaldo de teste in vitro de sensibilidade modificado, bem como a necessidade do uso responsável de antimicrobianos em animais de produção.(AU)

Diagnosis of mycobacteria in bovine milk: an overview

ABSTRACT Tuberculosis remains as the world’s biggest threat. In 2014, human tuberculosis ranked as a major infectious disease by the first time, overcoming HIV death rates. Bovine tuberculosis is a chronic disease of global distribution that affects animals and can be transmitted to humans by the consumption of raw milk, representing a serious public health concern. Despite the efforts of different countries to control and eradicate bovine tuberculosis, the high negative economic impact on meat and milk production chains remains, given the decreased production efficiency (approximately 25%), the high number of condemned carcasses, and increased animal culling rates. This scenario has motivated the establishment of official programs based on regulations and diagnostic procedures. Although Mycobacterium tuberculosis and Mycobacterium bovis are the major pathogenic species to humans and bovines, respectively, nontuberculous mycobacteria within the Mycobacterium genus have become increasingly important in recent decades due to human infections, including the ones that occur in immunocompetent people. Diagnosis of mycobacteria can be performed by microbiological culture from tissue samples (lymph nodes, lungs) and secretions (sputum, milk). In general, these pathogens demand special nutrient requirements for isolation/growth, and the use of selective and rich culture media. Indeed, within these genera, mycobacteria are classified as either fast- or slow-growth microorganisms. Regarding the latter ones, incubation times can vary from 45 to 90 days. Although microbiological culture is still considered the gold standard method for diagnosis, molecular approaches have been increasingly used. We describe here an overview of the diagnosis of Mycobacterium species in bovine milk.