Macrophage polarization in lymph node granulomas from cattle and pigs naturally infected with Mycobacterium tuberculosis complex.

Tuberculosis in animals is caused by members of the Mycobacterium tuberculosis complex (MTC), with the tuberculous granuloma being the main characteristic lesion. The macrophage is the main cell type involved in the development of the granuloma and presents a wide plasticity ranging from polarization to classically activated or pro-inflammatory macrophages (M1) or to alternatively activated or anti-inflammatory macrophages (M2). Thus, this study aimed to analyze macrophage polarization in granulomas from cattle and pig lymph nodes naturally infected with MTC. Tuberculous granulomas were microscopically categorized into four stages and a panel of myeloid cells (CD172a/calprotectin), M1 macrophage polarization (iNOS/CD68/CD107a), and M2 macrophage polarization (Arg1/CD163) markers were analyzed by immunohistochemistry. CD172a and calprotectin followed the same kinetics, having greater expression in late-stage granulomas in pigs. iNOS and CD68 had higher expression in cattle compared with pigs, and the expression was higher in early-stage granulomas. CD107a immunolabeling was only observed in porcine granulomas, with a higher expression in stage I granulomas. Arg1+ cells were significantly higher in pigs than in cattle, particularly in late-stage granulomas. Quantitative analysis of CD163+ cells showed similar kinetics in both species with a consistent frequency of immunolabeled cells throughout the different stages of the granuloma. Our results indicate that M1 macrophage polarization prevails in cattle during early-stage granulomas (stages I and II), whereas M2 phenotype is observed in later stages. Contrary, and mainly due to the expression of Arg1, M2 macrophage polarization is predominant in pigs in all granuloma stages.

Zoonotic tuberculosis in a high bovine tuberculosis burden area of Ethiopia.

Background: Tuberculosis (TB) is a major cause of ill health and one of the leading causes of death worldwide, caused by species of the Mycobacterium tuberculosis complex (MTBC), with Mycobacterium tuberculosis being the dominant pathogen in humans and Mycobacterium bovis in cattle. Zoonotic transmission of TB (zTB) to humans is frequent particularly where TB prevalence is high in cattle. In this study, we explored the prevalence of zTB in central Ethiopia, an area highly affected by bovine TB (bTB) in cattle. Method: A convenient sample of 385 patients with pulmonary tuberculosis (PTB, N = 287) and tuberculous lymphadenitis (TBLN, N = 98) were included in this cross-sectional study in central Ethiopia. Sputum and fine needle aspirate (FNA) samples were obtained from patients with PTB and TBLN, respectively, and cultures were performed using BACTEC™ MGIT™ 960. All culture positive samples were subjected to quantitative PCR (qPCR) assays, targeting IS1081, RD9 and RD4 genomic regions for detection of MTBC, M. tuberculosis and M. bovis, respectively. Results: Two hundred and fifty-five out of 385 sampled patients were culture positive and all were isolates identified as MTBC by being positive for the IS1081 assay. Among them, 249 (97.6%) samples had also a positive RD9 result (intact RD9 locus) and were consequently classified as M. tuberculosis. The remaining six (2.4%) isolates were RD4 deficient and thereby classified as M. bovis. Five out of these six M. bovis strains originated from PTB patients whereas one was isolated from a TBLN patient. Occupational risk and the widespread consumption of raw animal products were identified as potential sources of M. bovis infection in humans, and the isolation of M. bovis from PTB patients suggests the possibility of human-to-human transmission, particularly in patients with no known contact history with animals. Conclusion: The detected proportion of culture positive cases of 2.4% being M. bovis from this region was higher zTB rate than previously reported for the general population of Ethiopia. Patients with M. bovis infection are more likely to get less efficient TB treatment because M. bovis is inherently resistant to pyrazinamide. MTBC species identification should be performed where M. bovis is common in cattle, especially in patients who have a history of recurrence or treatment failure.

Causas de condenações em bovinos em um abatedouro com inspeção municipal de Minas Gerais / Causes of bovine condemnation in an slaughter under municipal inspection services in the Minas Gerais staction

A identificação de condenações em frigoríficos é importante para fornecer melhorias no processo produtivo e dados de possíveis fontes de contaminação na linha de abate. O objetivo foi identificar as causas de condenação de carcaças e vísceras bovinas, em um frigorífico de inspeção municipal em Minas Gerais, e discutir fatores que favorecem a ocorrência das mesmas. Foram analisados os registros de condenações no período de julho de 2017 a setembro de 2018. Do total de 3.723 animais abatidos, 101 (2,71%) apresentaram algum tipo de condenação. Foram condenados principalmente fígado, coração, fragmentos de caraças e carcaças inteiras. Abscessos hepáticos foram a causa mais frequente de condenação. Observamos que a frequência de descarte de carcaças e vísceras variaram entre os Serviços de Inspeção e estão relacionadas com a região estudada.

Sensibilidade in vitro a antimicrobianos de Corynebacterium pseudotuberculosis isolados de pequenos ruminantes da região Centro- Norte do Estado da Bahia / In vitro antimicrobial susceptibility of Corynebacterium pseudotuberculosis isolated from small ruminants from the North-Central region of the State of Bahia

Introdução: corynebacterium pseudotuberculosis é uma bactéria com importância em medicina veterinária por ser o agente etiológico da Linfadenite Caseosa (LC) em pequenos ruminantes. A doença leva a perdas econômicas devido a condenação de carcaças, diminuição de produtividade e redução do valor comercial do couro dos animais acometidos. O tratamento da doença é pouco eficaz, pois os agentes antimicrobianos não conseguem atravessar a barreira dos granulomas, mas tem sido proposta a inoculação intracapsular de antibióticos como tratamento e como medida de prevenção de contaminação ambiental. Objetivo: devido a esse fato, o presente estudo teve como objetivo avaliar a sensibilidade in vitro de isolados clínicos de Corynebacterium pseudotuberculosis isolados de animais da região Centro-Norte do estado da Bahia. Metodologia: foram retirados material de granulomas de 11 animais de uma fazenda na região acima mencionada, e as bactérias foram isoladas a partir desse material. Esses isolados então foram submetidos a metodologia de difusão em disco de ágar utilizando-se discos impregnados com doze antibióticos diferentes. Resultados: 90,9% (10/11) dos isolados foram resistentes a oxacilina. Com relação à sensibilidade, 100% dos isolados foram sensíveis a ampicilina, amicacina, amoxicilina, cefalotina, ceftriaxona, ciprofloxacina, enrofloxacina, neomicina e penicilina, enquanto 90,9% destes apresentaram sensibilidade a bactericina e doxiciclina. Conclusão: pode-se concluir que os isolados de C. pseudotuberculosis da região estudada apresentam sensibilidade a diversos antibióticos, o que pode facilitar na escolha de um antimicrobiano com menor toxicidade para fins de tratamento com inoculação intracapsular do medicamento.

Background: corynebacterium pseudotuberculosisis important bacteria in veterinary medicine because it is the Etiologic Agent of Caseous Lymphadenitis (CL) in small ruminants. The disease leads to economic losses due to condemnation of carcasses, productivity decrease and leather commercial value reduction of affected animals affected. The treatment of the disease is not very effective, because antimicrobial agents cannot cross the barrier of granulomas, but it has been proposed the intracapsular inoculation of antibiotics as treatment and as a measure of prevention of environmental contamination. Objective: due to this fact, the present study aimed to evaluate the in vitro susceptibility of clinical isolates of Corynebacterium pseudotuberculosis isolated from animals in the North Central region of the State of Bahia. Methodology: granuloma material were removed from 11 animals from a farm in the above mentioned area, and the bacteria were isolated from this material. These isolated bacteria were then subjected to disk diffusion method of agar using disks impregnated with twelve different antibiotics. Results: 90,9% (10/11) the isolated bacteria were resistant to oxacillin.With respect to sensitivity, 100% of the isolates bacteria were susceptible to amikacin, ampicillin, amoxicillin, cephalotin, ceftriaxone, ciprofloxacin, enrofloxacin, neomycin and penicillin, while 90.9% of these showed sensitivity to bacitracin and doxycycline. Conclusion: it can be concluded that the isolated bacteria of C. pseudotuberculosis from the region studied present sensitivity to different antibiotics, which can facilitate the choice of a less toxic antimicrobial for treating with intracapsular inoculation of the medicine.

Prevalence of tuberculous lymphadenitis in slaughtered cattle in Eastern Cape, South Africa.

OBJECTIVE: To detect the prevalence of Mycobacterium tuberculosis complex (MTBC) in the lymph nodes of slaughtered cattle collected from selected abattoirs in Eastern Cape Province of South Africa. METHODS: A total of 376 lymph nodes were collected from slaughtered cattle over a period of 12 months. Certain characteristics (sex, age, body condition score, and breed) were observed to be associated with MTBC among slaughtered cattle. Collected samples were cultured and tested for acid-fast bacilli (AFB). DNA was isolated, purified, and quantified using a spectrophotometer. Quantified DNA was confirmed to be MTBC by multiplex PCR targeting two genes (IS6110 and mpb64). RESULTS: Of the 376 collected lymph nodes, 182 were positive when tested by Ziehl-Neelsen stain and 162 were confirmed positive for MTBC by PCR. MTBC was isolated from lymph nodes with nodular lesions (72.8%, 118/162) and inflamed lymph nodes (27.1%, 44/162). All detected MTBC isolates were positive for region of deletion 1 (RD1). No isolate was detected to have Mycobacterium bovis bacille Calmette-Guérin (BCG). However, 3.1% had M. bovis and 96.9% had M. tuberculosis. CONCLUSIONS: The presence of live Mycobacterium strains in slaughtered cattle poses a health risk to beef consumers and abattoir workers.

Systemic infection of Mycobacterium avium subspecies hominissuis and fungus in a pet dog.

A 3-year-old neutered female poodle with a long history of dermatophytic skin disease was presented with lethargy, anorexia and progressive weight loss. Abdominal ultrasonography revealed markedly enlarged mesenteric lymph nodes and multiple hypoechoic foci in the spleen. Cytology of the mesenteric lymph nodes and spleen showed granulomatous inflammation with fungal organisms and negatively stained intracytoplasmic bacterial rods consistent with Mycobacteria spp. Based on culture, multiplex polymerase chain reaction and sequence analysis, the bacterium was identified as Mycobacterium avium subspecies hominissuis. Despite treatment with antibiotics, the dog's condition deteriorated, and it died approximately 3 weeks after first presentation.

Evaluation of the efficiency of nested q-PCR in the detection of Mycobacterium tuberculosis complex directly from tuberculosis-suspected lesions in post-mortem macroscopic inspections of bovine carcasses slaughtered in the state of Mato Grosso, Brazil.

Bovine tuberculosis (BTB) is a zoonotic disease caused by Mycobacterium bovis, a member of the Mycobacterium tuberculosis complex (MTC). The quick and specific detection of this species is of extreme importance, since BTB may cause economic impacts, in addition to presenting imminent risks to human health. In the present study a nested real-time PCR test (nested q-PCR) was used in post-mortem evaluations to assess cattle carcasses with BTB-suspected lesions. A total of 41,193 cattle slaughtered in slaughterhouses located in the state of Mato Grosso, were examined. Of the examined animals, 198 (0.48%) showed BTB-suspected lesions. M. bovis was isolated in 1.5% (3/198) of the samples. Multiplex-PCR detected MTC in 7% (14/198) of the samples. The nested q-PCR test detected MTC in 28% (56/198) of the BTB-suspected lesions, demonstrating higher efficiency when compared to the multiplex-PCR and conventional microbiology. Nested q-PCR can therefore be used as a complementary test in the national program for control and eradication of bovine tuberculosis.

Pre-multidrug-resistant Mycobacterium tuberculosis Beijing strain associated with disseminated tuberculosis in a pet dog.

Resistance to isoniazid, ethambutol, and streptomycin was detected in a Mycobacterium tuberculosis strain, belonging to the Beijing family lineage, isolated from two nodule exudates of a Yorkshire terrier with generalized tuberculosis. This report alerts medical practitioners to the risk of dissemination of pre-multidrug-resistant tuberculosis (preMDR-TB) through exposure to M. tuberculosis-shedding pets.

Diagnosis of Mycobacterium avium complex in granulomatous lymphadenitis in slaughtered domestic pigs.

The aim of this study was to compare two diagnostic methods for the detection of Mycobacterium avium complex (MAC) infection in lymph nodes with granulomatous lymphadenitis from slaughtered domestic pigs. Fifty affected lymph nodes were collected from 50 pigs and examined microscopically and by polymerase chain reaction (PCR). Microscopically, granulomatous lesions were observed in 92% of the samples, consisting mostly of central necrosis (78%) with dystrophic calcification (46%) and associated with inflammatory infiltration by epithelioid giant cells, lymphocytes, neutrophils (92%), eosinophils (60%) and Langhans-type cells (70%). In 64% of the lesions, a capsule of connective tissue was found. Acid-fast bacilli were observed in all cases. PCR detected DNA from Mycobacterium spp. in 82% (41/50) of the lymph nodes. MAC was confirmed in 58% (24/41) and M. avium avium/silvaticum subspecies in 39% (16/41). The results of this study suggest that combined histopathology and PCR of lymph nodes are useful in the diagnosis of granulomatous lymphadenitis in slaughtered pigs.

Outbreak of tuberculosis caused by Mycobacterium bovis in golden Guernsey goats in Great Britain.

An outbreak of caprine tuberculosis caused by Mycobacterium bovis was disclosed in June 2008, affecting goats of the golden Guernsey breed kept on 10 separate smallholdings in south-west Wales and the west of England. Following the initial diagnosis at postmortem examination, 30 goats that reacted positively to the single intradermal comparative cervical tuberculin (SICCT) test, together with five in-contact animals, were euthanased and subjected to postmortem examination and mycobacterial culture. Spoligotyping and variable number tandem repeat analysis of isolates showed that they were all of the same genotype, endemic to south-west Wales. Retrospective movement tracings identified a goat herd in south-west Wales, by then completely dispersed, as the probable common source of infection. There was a perfect correlation between the SICCT test and culture results in all slaughtered goats. Grossly visible tubercular lesions were observed at postmortem examination in all but one reactor.

IS1245 RFLP analysis of strains of Mycobacterium avium subspecies hominissuis isolated from pigs with tuberculosis lymphadenitis in Portugal.

From the end of 2004 increasing numbers of tuberculosis-like lesions were detected in regular meat inspections of pigs at regional abattoirs. Mycobacterium avium subspecies hominissuis was identified as the cause of this outbreak of tuberculosis lymphadenitis by IS901 and IS1245 duplex PCR and FR300 PCR. In order to detect the source and route of infection, IS1245 restriction fragment length polymorphism (RFLP) analysis was used to type 54 strains (50 from pigs, one from a cow, two from wild boar and one from peat) isolated in 12 different regions of Portugal. A total of 51 different patterns were obtained with most strains giving a high IS1245 copy number (between seven and 27) with a complex RFLP pattern. Twenty-three strains were gathered in 11 clusters, defined as having at least 80 per cent similarity. Identical isolates were obtained from one herd in Lisbon (two isolates), one herd in Setúbal (two isolates) and in two isolates from different herds in the same region. No distinctive geographical clustering was found among the 54 strains. These results indicate that strains of M hominissuis are very heterogeneous and widespread in pigs in Portugal. No major common source of the infection was identified.

Tuberculosis in ruminants: characteristics of intra-tonsilar Mycobacterium bovis infection models in cattle and deer.

Mycobacterium bovis infection produces tubercular lymphadenitis in the head lymphatics of cattle and deer, in addition to pulmonary disease. A low-dose intra-tonsilar infection model that establishes tuberculosis (Tb) lymphadenitis in cattle and deer is characterised in this study. Intra-tonsilar infection of red deer (500 cfus of M. bovis) was monitored longitudinally at 6-week intervals over a period of 23 weeks. Lesion characteristics, bacteriological and immunological parameters were assessed, and compared against those observed in cattle at 20 weeks post-infection, where the latter were infected with 500 or 5000 cfus of M. bovis. Intra-tonsilar inoculation of M. bovis established infection in >90% of deer and cattle, with lesion frequencies at the draining sentinel lymphatic site (left medial retropharyngeal node) of 68-86% and tissue bacterial burdens >3.5 logs/g of tissue, the tonsil being a major site of M. bovis persistence in deer only. Mineralisation occurred at lesion sites in both species in the later stages (18-23 weeks) of infection, with extensive coarse mineralisation observed mainly in cattle. The severity of infection or disease in cattle that received the higher or lower dose of M. bovis did not differ markedly. Pathogen-induced cellular immune response (lymphocyte transformation) and humoral responses (IgG and IgG(1) anti-mycobacterial antibodies) were recorded in both species, and the magnitude of these was noticeably amplified by skin tuberculin testing. IgG(1) antibodies were detectable within 6 weeks post-inoculation in deer and could be associated with early detection of lymphadenitis. Deer and cattle show similar levels of susceptibility to M. bovis infection.

Impact of sawdust and wood shavings in bedding on pig tuberculous lesions in lymph nodes, and IS1245 RFLP analysis of Mycobacterium avium subsp. hominissuis of serotypes 6 and 8 isolated from pigs and environment.

Among 25,027 slaughter pigs raised in two farms, tuberculous lesions were detected in the lymph nodes of 898 (3.6%) of them. Tuberculous lesions were most commonly found in the mesenteric (601; 2.4%) and head (451; 1.8%) lymph nodes. Mycobacteria were isolated from 49 of 120 randomly selected mesenteric, head and bronchial lymph nodes with diagnosed tuberculosis originating from both farms. Forty six Mycobacterium avium subsp. hominissuis, one M. chelonae and two M. fortuitum isolates were found in the lymph nodes of pigs. No statistically significant difference was detected between farms A and B for isolation rates of mycobacteria from the lymph nodes of pigs and their species composition. To investigate the source of the pigs' infections, culture examinations of 117 samples from the external environment were performed. Mycobacteria were isolated from 25 samples from the external environment (21.4%). Mycobacterial isolates were also detected in eleven (91.7%) and two (16.7%) of 12 used sawdust and 12 of non-used (fresh) sawdust samples, respectively. None of 12 wood shavings was culture-positive. Twelve of 13 sawdust isolates were classified as M. a. hominissuis of serotypes 6 and 8 and genotype IS901- and IS1245+; the remaining isolate was classified as species M. fortuitum. Other conditionally pathogenic mycobacteria were only isolated from 12 of the remaining 81 samples from the external environment (excluding bedding). A total of eight isolates (two pig and six sawdust samples originating from farms A and B) were examined by IS1245 restriction fragment length polymorphism (IS1245 RFLP) analysis. These isolates produced five distinct IS1245 RFLP types with more than 20 bands. Based on identical IS1245 RFLP types of one pig isolate and two isolates of used sawdust from farm A, we have concluded that contaminated sawdust was the source of mycobacterial infection for pigs in our study.

Milk containing Mycobacterium bovis as a source of infection for white-tailed deer fawns (Odocoileus virginianus).

SETTING: White-tailed deer represent the first wildlife reservoir of Mycobacterium bovis in the United States. The behavior of does with nursing fawns provides several potential mechanisms for disease transmission. Little information exists concerning transmission between doe and fawn, specifically transmammary transmission. OBJECTIVE: Determine if fawns can become infected by ingestion of milk replacer containing M. bovis, thus simulating transmission from doe to fawn through contaminated milk. DESIGN: Seventeen, 21-day-old white-tailed deer fawns were inoculated orally with 2 x 10(8) CFU (high dose, n=5), 2.5 x 10(5) to 2.5 x 10(6) CFU (medium dose, n=5), and 1 x 10(4) CFU (low dose, n=5) of M. bovis in milk replacer. Dosages were divided equally and fed daily over a 5-day period. Positive control fawns (n=2) received 1 x 10(5) CFU of M. bovis instilled in the tonsillar crypts. Fawns were euthanized and examined 35-115 days after inoculation and various tissues collected for bacteriologic and microscopic analysis. RESULTS: All fawns in the tonsillar, high oral and medium oral dose groups developed generalized tuberculosis involving numerous organs and tissues by 35-84 days after inoculation. Three of five fawns in the low-dose oral group had tuberculous lesions in the mandibular lymph node, and one of five had lesions in the medial retropharyngeal lymph node when examined 115 days after inoculation. CONCLUSION: White-tailed deer fawns can become infected through oral exposure to M. bovis. Therefore, the potential exists for fawns to acquire M. bovis while nursing tuberculous does.

Tuberculosis, caused by Mycobacterium bovis, in a kudu (Tragelaphus strepsiceros) from a commercial game farm in the Malelane area of the Mpumalanga Province, South Africa.

Tuberculosis, caused by Mycobacterium bovis, was diagnosed for the first time, in a kudu cow from a commercial game ranch in the Malelane area of the Mpumalanga Province close to the Kruger National Park. This diagnosis has important implications for the eradication of the disease in commercial and communal livestock in the area. Kudus are considered to be a potential maintenance host and, because of discharging fistulae in the parotid area where the lymph nodes are commonly infected, they have the potential of disseminating bacteria over wide areas. Cognisance should be taken of the presence of tuberculosis in a species other than domesticated cattle in this area and its implications for the control of tuberculosis in cattle.

Tuberculosis in kudus (Tragelaphus strepsiceros) in the Kruger National Park.

Five kudus (Tragelaphus strepsiceros), three bulls and two cows, within the Greater Kruger National Park complex, were diagnosed with generalized tuberculosis caused by Mycobacterium bovis. The lesions seen in these animals were similar to those previously reported in kudus and included severe tuberculous lymphadenitis of the nodes of the head and neck (that resulted in noticeable uni- or bilateral swelling beneath the ear), thorax, and the mesentery. All the animals also suffered from severe granulomatous pneumonia. The lesions in the lungs were more severe cranially and had a miliary distribution elsewhere in the lungs. Based on the DNA patterns of the M. bovis isolates, at least some of these kudus were infected with strains commonly present in tuberculous buffaloes, lions, cheetahs, and baboons in the Park whereas other strains from these kudus were quite different and may reflect another source of infection. The presence of tuberculous kudus in the Park is expected to complicate control measures that may be instituted to contain or eradicate the disease in the Park.

[Tuberculosis lymphadenitis in slaughtered swine from the State of São Paulo, Brazil: microscopic histopathology and demonstration of mycobacteria]. / Linfadenites tuberculóides em suínos abatidos no estado de São Paulo, Brasil: aspectos macroscópicos histopatológicos e pesquisa de micobactérias.

INTRODUCTION: As the occurrence of tuberculosis lymphadenitis in swine constitutes a public health risk, especially in immunosuppressed individuals, the distribution of tuberculoid lesions and the presence of microbacteria in lymph nodes and hepatic and muscular tissue in swine for slaughter, in the State of S. Paulo, SP (Brazil), in the period from 1993 to 1994, were studied. MATERIAL E METHOD: Tuberculous lesions in 60 carcasses of swine, slaughtered at abattoirs during the period 1993-1994, were studied. When macroscopic lesions were observed, a representative sampling of lymph nodes, hepatic and muscular tissues (masseter and diaphragm) were examined using histopathological and microbacterial isolation techniques. RESULTS: The macroscopic lesions were found predominantly in mesenteric lymph nodes. The MAI Complex (M.avium-intracellulare) was isolated mainly in the carcass group that showed lesions and was not found in the control group. Microorganisms of the MAI Complex were not isolated from hepatic and muscular tissues. CONCLUSION: No clear relationship between the type of mycobacteria isolated and the macroscopic lesions observed during the carcass inspection was found.

An abattoir study of tuberculosis in a herd of farmed elk.

The purpose of this study was to examine the prevalence and distribution of grossly visible lesions of tuberculosis in a herd of 344 North American elk (Cervus elaphus) depopulated during a three-month period in 1991. Abattoir inspection detected mycobacterial lesions in 134 (39.8%) of the 337 animals received for slaughter. The prevalence of lesions increased with the age of the animals. Lesions were predominantly suppurative rather than caseous, and mineralization was less evident than in tuberculous lesions in cattle. Lesions occurred predominantly in lymph nodes, and lungs were the only organs in which mycobacterial lesions were found. The distribution of lesions suggested that aerosol transmission was the most significant means of spread of the disease within the herd. Giant liver flukes (Fascioloides magna) were observed in approximately 80% of the adult elk.